CN106421322A - Traditional Chinese medicine composition capable of preventing and treating infectious bronchitis of chickens and preparation method of traditional Chinese medicine composition - Google Patents
Traditional Chinese medicine composition capable of preventing and treating infectious bronchitis of chickens and preparation method of traditional Chinese medicine composition Download PDFInfo
- Publication number
- CN106421322A CN106421322A CN201611017765.1A CN201611017765A CN106421322A CN 106421322 A CN106421322 A CN 106421322A CN 201611017765 A CN201611017765 A CN 201611017765A CN 106421322 A CN106421322 A CN 106421322A
- Authority
- CN
- China
- Prior art keywords
- chinese medicine
- medicine composition
- chicken
- flowered skullcap
- preventing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
- A61K36/539—Scutellaria (skullcap)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
- A61K36/233—Bupleurum
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/35—Caprifoliaceae (Honeysuckle family)
- A61K36/355—Lonicera (honeysuckle)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/63—Oleaceae (Olive family), e.g. jasmine, lilac or ash tree
- A61K36/634—Forsythia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/74—Rubiaceae (Madder family)
- A61K36/744—Gardenia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8966—Fritillaria, e.g. checker lily or mission bells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Neurosurgery (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention provides a traditional Chinese medicine composition capable of preventing and treating infectious bronchitis (IB) of chickens. The traditional Chinese medicine composition is prepared from the following crude drugs in parts by weight: radix scutellariae, fructus forsythiae, fructus gardeniae, honeysuckle flower, cablin potchouli herb, radix bupleuri and bulbus fritillariae thunbergii. The composition provided by the invention has unique effects in the aspects of animal growth prompting and IB prevention and treatment. The traditional Chinese medicine composition is safe, has no toxin and side effect, has good treatment effect and low cost and does not influence egg laying; the composition provided by the invention has cough relieving, heat clearing, pain easing and inflammation resisting effects; the observation of treatment effect and clinical verification of the chickens artificially infected with IBV (Infectious Bronchitis Virus) proves that after the preparation provided by the invention is eaten by the chickens, symptoms are alleviated, the mental status is improved, the feed intake is increased, cough and asthma symptoms disappear and the like; only a few of the chickens have symptoms of shaking heads and sneezing. The composition provided by the invention can be used for effectively alleviating sick chicken clinical symptoms caused by the IBV; the total effective rate reaches to 94 percent or more, and the healing rate and the effective rate are higher than those of a positive control group. The composition can be applied to the prevention and treatment of IB in a large scale.
Description
Technical field
The invention belongs to Chinese medicine compound prescription field, is related to a kind of Chinese medicine composition of preventing and treating infectious bronchitis of chicken and its system
Preparation Method.
Background technology
Infectious bronchitis of chicken (Infectious bronchitis, IB) is by IBV
A kind of acute high degree in contact respiratory infectious disease that (Infectious bronchitis virus, IBV) causes, because passing
The rarely seen chicken infection of metachromia bronchitis early stage, so cock infective bronchitis.Clinical manifestation is for coughing, sneezing, kidney
Infringement, egg production are reduced, Egg Quality declines etc..Avian infectious bronchitis virus has that speed of mutation is fast, serotype is complicated,
The characteristics of Disease Clinical diversity ratio is larger.The disease can infect the chicken in any period, cause delayed growth, lose weight and extremely
Die, cause the secondary infections such as Mycoplasma mixed infection and colibacillosis, increase chicken group's The dead quantity, bring sternly to plant
The loss of weight.At present for infectious bronchitis of chicken there is no effective treatment method, mainly protected using epidemic prevention means
Shield chicken group, but as the serum virus type is more, has failure during clinical vaccine immunity, and virus can often produce new variation
Strain, causes vaccine immunity risk of failure high, and poor to IB preventive effects, antibiotic easily produces drug resistance in addition, easily residual etc..Right
Symptomatic treatment, traditional Chinese medicine is mainly adopted to be mainly pulvis at present in clinical onset chicken group, preparation process is simple, and product is controllable
Property is poor, bad when good during Clinical practice effect.Infectious bronchitis of chicken is broadly divided into breathing pattern and kidney type, but preinfection
Mainly breathing pattern, with the prolongation of the course of disease, is developing progressively kidney type etc..Therefore, controlling for infectious bronchitis of chicken
Treat beforehand control more crucial.Therefore in the urgent need to a safe and effective medicine.
Content of the invention
It is an object of the invention to provide a kind of prevention effect significantly, the preventing and treating infectious bronchitis of chicken that do not affect to lay eggs
Chinese medicine composition and preparation method thereof.
To achieve these goals, the technical solution adopted in the present invention is as follows:
A kind of Chinese medicine composition of preventing and treating infectious bronchitis of chicken, it is characterised in that the Chinese medicine composition is by following weight
The bulk drug of number is prepared from:100-200 part roots of large-flowered skullcap, 100-200 part capsules of weeping forsythia, 100-200 part cape jasmines, 50-150 part gold and silver
Flower, 100-200 part Pogostemon cablin, 10-100 part radix bupleuri, 50-150 part fritillaria thunbergiis.
Preferably, the Chinese medicine composition is prepared from by the bulk drug of following parts by weight:120-180 part roots of large-flowered skullcap,
120-180 part capsules of weeping forsythia, 120-180 part cape jasmines, 60-140 part honeysuckles, 120-180 part Pogostemon cablin, 20-80 part radix bupleuri, 60-140
Part fritillaria thunbergii.It is furthermore preferred that what the Chinese medicine composition was prepared from by the bulk drug of following parts by weight:140-160 part roots of large-flowered skullcap,
140-160 part capsules of weeping forsythia, 140-160 part cape jasmines, 80-120 part honeysuckles, 140-160 part Pogostemon cablin, 40-60 part radix bupleuri, 80-120
Part fritillaria thunbergii.
Further, the Chinese medicine composition of described preventing and treating infectious bronchitis of chicken, it is characterised in that:The Chinese medicine group
Compound is powder, capsule, oral liquid, granule, parenteral solution or tablet.
Present invention also offers a kind of preparation method of the Chinese medicine composition of preventing and treating infectious bronchitis of chicken, its feature exists
As follows in step:The root of large-flowered skullcap, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and the fritillaria thunbergii of recipe quantity is weighed, fine powder is ground into,
80-100 mesh sieves are crossed, is mixed, packing is obtained powder.
Present invention also offers a kind of preparation method of the Chinese medicine composition of preventing and treating infectious bronchitis of chicken, its feature exists
As follows in step:Weigh the root of large-flowered skullcap, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and the fritillaria thunbergii of recipe quantity, the medicine in addition to the root of large-flowered skullcap
Material first adds the water immersion 0.5-3 hours that 8-12 times of prescription total weight measures, and decocts 2-3 time, and wherein the root of large-flowered skullcap treats that head liquid of boiling medicine boils
Under afterwards, each 1-2 hours, collecting decoction are evaporated to thick paste, and vacuum drying is spray-dried, and is ground into fine powder, crosses 100
Mesh sieve, adds appropriate amount of auxiliary materials, mixes, prepares capsule, granule or tablet.
Present invention also offers a kind of preparation method of the Chinese medicine composition of preventing and treating infectious bronchitis of chicken, its feature exists
As follows in step:Weigh the root of large-flowered skullcap, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and the fritillaria thunbergii of recipe quantity, the medicine in addition to the root of large-flowered skullcap
Material first adds the water immersion 0.5-3 hours that 8-12 times of prescription total weight measures, and decocts 2-3 time, and wherein the root of large-flowered skullcap treats that head liquid of boiling medicine boils
Under afterwards, each 1-2 hours, collecting decoction are evaporated to relative density 1.05~1.10 (50 DEG C), plus ethanol reaches alcohol content
50-70%;10 DEG C of 24-72 hours arranged below, leaching supernatant reclaim ethanol to without alcohol taste, add preservative, be diluted with water
To oral liquid 0.6~1.0g containing primary crude drug of every 1mL, filtration is filling, and sterilizing obtains final product oral liquid formulations.
Preferably, the preparation method of the Chinese medicine composition of preventing and treating scouring, it is characterised in that:Extraction technique is:
Add water 10 times and measure, decoct 3 times, for the first time 2 hours, second, each 1 hour for the third time.
Preferably, impurity removal process parameter is:Extract is concentrated into relative density 1.05~1.10 (50 DEG C), plus ethanol makes to contain
Alcohol amount is up to 60%.
The invention also discloses described Chinese medicine composition is used for treating preparing or prevents in infectious bronchitis of chicken
Application.Using method:The powder is added in feed according to 0.5~1.0% weight ratio, mixes thoroughly, be used in conjunction 5-7 days, you can
Infectious bronchitis of chicken is effectively prevented and treated.
The Chinese medicine composition of the present invention is mainly by groups such as the root of large-flowered skullcap, the capsule of weeping forsythia, honeysuckle, cape jasmine, Pogostemon cablin, radix bupleuri, fritillaria thunbergiis
Into being that warm disease is set from the beginning of, heat in both exterior and interior caused by affection of exogenous wind-heat, IBV can be infected etc. and to be prevented and treated..
Root of large-flowered skullcap bitter cold in side and gas is thin, main enter the part of the body cavity above the diaphragm housing the heart and lungs, the kind fire-evil that lets out in lung controls dampness-heat in upper jiao, treats hot disease on diaphragm, except the circulation of vital energy in the wrong direction in the heart,
For monarch drug in a prescription.Honeysuckle is sweet cold, wind-dispelling heat-dissipating, thoroughly evil removing toxic substances, inclined clearing lung-heat fire;Capsule of weeping forsythia hardship is slightly cold, light weight and float, rush down the six channels stagnated fire;
Cape jasmine is bitter, trembles with fear.Kind removing the heart-fire, lung, three burnt tired fire.Three medicine phases are closed, and it is ministerial drug to evacuate in table wind-heat.The pungent tepor of Pogostemon cablin, aromatization
Lung channel turbid, and stop taste discomfort vomiting, deliver solve whole body summer heat;Radix bupleuri is pungent, bitter, is slightly cold, and evacuation is brought down a fever, soothing the liver solution
Strongly fragrant, rise and lift yang-energy;Two medicines are shared, and assistant helps the heat of the clear whole body of ministerial drug, and the preventing or arresting vomiting that can dehumidify.Fritillaria thunbergii has the property of sweet cool moistening lung,
Can moisten the lung and nourish yin, again can preventing phlegm from forming and stopping coughing, for making medicine;All medicine phases are closed, altogether long memorial dissipate wind-heat, reducing pharyngeal swelling, the effect of removing heat from the lung to relieve cough.
Prove through the clinical treatment IBV respiratory tract disease symptom that causes of infection and live application study, we with
Dispelling wind and heat from the body, reducing pharyngeal swelling, removing heat from the lung to relieve cough are the rules for the treatment of, are primarily directed to down in spirits, poor appetite that IBV causes, productivity
Can decline, particularly get rid of nose, sneeze, gradually develop into cough, have difficulty in breathing, tracheal sound, tracheitis, bronchitis, or even
Pulmonary congestion, oedema, the breathing problem based on different degrees of pneumonia.
Chinese veterinarian thinks that avian infectious bronchitis virus mainly passes through respiratory tract infection in body deficiency of vial QI, by
In respiratory tract infection virus after cause to accumulate in heat toxin, cause sputum obstruction tracheae, make chicken occur body temperature raise, cold, whole body without
The symptoms such as power, laziness, cough, asthma.The present invention for IB clinical symptoms, treat using Chinese veterinarian is dialectical, with dispelling wind and heat from the body,
Reducing pharyngeal swelling, removing heat from the lung to relieve cough are the rules for the treatment of.The root of large-flowered skullcap, capsule of weeping forsythia etc. are acted on anti-IBV;Cape jasmine Ultramicro-powder Carrageenan causes mouse foot
Swelling has obvious inhibiting effect, paraxylene to cause mice auricle swelling to have obvious inhibiting effect, Dichlorodiphenyl Acetate to cause mouse writhing to hide
Phase is obviously prolonged effect, has synergy to hypnosis of pentobarbital sodium of subthreshold dose;Three is used in conjunction and can strengthen heat-clearing
The effect of removing toxic substances;Pogostemon cablin is removed dampness by means of aromatics, and can help remove body foul smell, and patchouli oil drops can endotoxin-induced fever rabbit body temperature
Low, with refrigeration function;Radix bupleuri is diaphoretic medicine, can dissipate in heat, radix bupleuri to the rat that generates heat caused by yeast, with certain antipyretic
Effect;Fritillaria thunbergii is clearing heat and eliminating phlegm medicine, can resolving sputum saliva;Peimine is with peiminine to chemical stimulation mouse, machinery thorn
Swash to draw and cough cavy and electro photoluminescence is drawn and coughs cat and have antitussive effect and central inhibitory action.
Medicine composition and analysis:
The root of large-flowered skullcap:Bitter, cold in nature.Return lung, courage, spleen, large intestine, small intestinl channel.Indication:Heat-clearing and damp-drying drug, purging intense heat and detonicating, only
Blood, antiabortive.For damp-warm syndrome, heat temperature evil of vomitting uncomfortable in chest, damp and hot feeling of fullness, rush down dysentery, jaundice, cough with lung heat, hyperpyrexia polydipsia, blood-head tells nosebleed,
Carbuncle sore tumefacting virus, fetal irritability.《Do not record》:Treat phlegm hot, pathogenic heat in the stomach, little eilema, disappear paddy, and sharp small intestine, woman's blood are closed, and drenches under dew
Blood, abdominal pain in children.《Haigoushen》:Heat toxin can be controlled, hectic fever due to yin, fevers and chills alternate, stomach are unfavorable, break and stop up gas, control five pouring, make us a surname freely,
Go to joint unhappy, antipyretic thirsty, hot abdominal pain is controlled, trusted subordinate's heavily fortified point is swollen.《The southern regions of the Yunnan Province book on Chinese herbal medicine》:Up purte the white, the descending bladder that rushes down are fiery,
(controlling) man five is drenched, woman's bleeding, menstruation regulating heat-clearing, and tire has burning hot uneasiness, and clear tire heat, except the six channels excess fire real heat.
Radix bupleuri:Bitter, cold nature.Return liver, gallbladder channel.Indication:In inducing sweat, soothing the liver, rising Yang.For cold, fever,
Fevers and chills alternate, distending pain in the chest and hypochondrium, irregular menstruation, sub-official prolapses, prolapse of the anus.《This warp》:Gas is tied in main trusted subordinate's stomach, and diet is gathered, and is trembled with fear
Heat evil gas, pushes away Chen Zhixin.《Do not record》:Except dysphoria with smothery sensation under the typhoid fever heart, all phlegm thermojunction realities are evil inverse in the heart, and faint breath between five Tibetan, large intestine stop
Product, water swelling, and arthritis with fixed pain caused by dampness contraction.《Haigoushen》:Zhi Relao restless bone pains, hot gas, shoulder and back ache declare smooth vim and vigour, tired thin thin;Main
Lower gas helps digestion, and when main, inside and outside disease, heat is not understood, and singly boils clothes.《Japan hanako materia medica》:General debility is mended, relieving restlessness is only frightened, beneficial gas power, dissolving phlegm
Only cough, embellish lung adds smart mends marrow, and day row temperature disease is fanatical weary exhausted, and chest side of body gas is full, forgetful.《The southern regions of the Yunnan Province book on Chinese herbal medicine》:Typhoid fever inducing sweat and dispelling exogenous evils will
Medicine, moves back the six channels heat symptoms caused by an exopathgen and comes and goes, numbness impotence, and except liver man heat symptoms caused by an exopathgen, consumptive disease heat, the gas of the inverse knot of row Liver Channel, only left flank irritability pain, controls married woman's blood
Heat burns warp, can adjust menstruation.
Honeysuckle:Sweet, cold in nature.Return lung, the heart, stomach.Indication:Clearing heat and detoxicating, wind-heat dissipating.For carbuncle swells malignant boil
Sore, larynx numbness, erysipelas, toxic-heat and blood stasis, anemopyretic cold, warm disease heating.《The southern regions of the Yunnan Province book on Chinese herbal medicine》:Heat-clearing, solves all sores, ulcer carbuncle on the back, red stream
Scrofula.《The sward property of medicine is standby will》:The ulcer that can disappear serious case of furuncle, only dysentery, washes infantile malnutrition due to digestive disturbances or intestinalparasites sore, goes skin blood-head.《Chongqing hall random notes》:In clear network
Wind fire is damp and hot, the dirty filthy heresy of the warm epidemic disease of solution, the floating more wind sun of breath liver and gall, controls convulsion and faints all diseases of epilepsy.
Fritillaria thunbergii:Bitter, cold in nature.Return lung, the heart channel of Hang-Shaoyin.Indication:Heat-clearing dissipating bind, preventing phlegm from forming and stopping coughing, for wind-heat affecting lung, phlegm
Fire cough, lung carbuncle, acute mastitis, scrofula, sore.《Book on Chinese herbal medicine is just》:Control greatly lung carbuncle lung to wither, cough and asthma, spit blood, bleeding from five sense organs or subcutaneous tissue most drops mental disorder, be apt to open
Pent-up, relieve the pain pain, and dissipate-swelling is full, clearing away the liver-fire, bright knowledge, except when gas dysphoria with smothery sensation, jaundice drenches and closes, excessive blood of having blood in stool;Relieving heat toxin, kill parasites and
Treatment larynx numbness, scrofula, acute mastitis carbuncle on the back, all large carbuncle pyogenic infections, damp and hot evil sore, anal fistula, metal-inflicted wound bleeding, fiery sore pain, than Bulbus Fritillariae Cirrhosae,
The work(of clear drop, not less than several times.《The new compilation of materia medica》:Wind phlegm is felt when going.《Detailed outline is picked up any lost article from the road》:The sharp phlegm of removing toxic substances, drives a surname's lung qi, all lung man folders
Wind fire has phlegm person preferably this.《Shandong Chinese herbal medicine handbook》:Clearing lung and eliminating phlegm, relieving haperacidity, removing toxic substances.Cure cold cough, and acid regurgitation of having a stomachache, carbuncle poison are swollen
Bitterly.
The capsule of weeping forsythia:Bitter, cool in nature.Enter the heart, liver, gallbladder channel.Indication:Heat-clearing, removing toxic substances, dissipating bind, detumescence.Control warm, erysipelas,
Macula, large carbuncle pyogenic infections, scrofula, urine leaching closes.《This warp》:Main fever and chills, mouse impotence, scrofula, Carbuncle swells dislikes sore, goiter and tumor, knot heat.《The property of medicine
By》:Main tonneau five drenches, urinary obstruction, except heart man visitor's heat.《Japan hanako materia medica》:Logical small intestine, apocenosis.Boil is controlled, the moon is led in pain relieving
Through.Li Gao:Dissipate all menses knot gas to gather;Detumescence.
Cape jasmine:Bitter, cold in nature.The thoughts of returning home, lung, tri-jiao channel.Indication:Purging intense heat relieving restlessness, reducing fever and causing diuresis, removing pattogenic heat from the blood and toxic material from the body.With
In Vexed, jaundice urine is red, blood strangury and dry pain, blood-head tells nosebleed, red eye, swell pain, pathogenic fire,toxin and furuncles.《This warp》:Perverse trend in main five, stomach
Middle hot gas, flushing, wine blister nose, white leprosy, red leprosy, sore.《Do not record》:Treat mesh heat also pain, the big heat of the chest heart, intestine and small intestine, vexation
Vexed, pathogenic heat in the stomach gas.《Medicine origin》:The heart channel of Hang-Shaoyin visitor's heat is treated, except agitation, part of the body cavity above the diaphragm housing the heart and lungs abnormal heat is gone, is controlled wind.Zhu Zhenheng:Rush down three burnt fire, clearing stomach
Gastral cavity blood, controls pyretic epigastralgia, antipyretic strongly fragrant, row knot gas.《Detailed outline》:Control haematemesis, bleeding from five sense organs or subcutaneous tissue, bloody flux, lower blood, blood strangury, traumatic blood stasis, and wound
Cold labor is multiple, pyretic headache, hernia, burn.
Pogostemon cablin:Aerial part for labiate Pogostemon cablin Pogostemoncablin (Blanco) Benth..Property
Tepor, taste is pungent.Indication:Eliminate dampness with aromatics, whet the appetite preventing or arresting vomiting, delivers relieving summer-heat.For turbi damp obstructing in middle-JIAO, the vomiting of gastral cavity ruffian, summer-heat and damp burnout,
Not easypro, cold-dampness uncomfortable in chest closes heat, stomachache vomiting and diarrhoea, nasosinusitis headache.《Do not record》:Treat geomantic omen poison swollen, remove bad odor, treat cholera, pained.《This
Sketch is passed through》:Control taste and tell inverse, be the medicine that most wants.《Pearl sac》:Benefit defends gas, beneficial stomach Qi, enters diet, controls again and tells inverse cholera.《Soup
Liquid book on Chinese herbal medicine》:The fast gas of middle benefit gas.The deficiency syndrome of the lung has cold, part of the body cavity above the diaphragm housing the heart and lungs heap soil or fertilizer over and around the roots heat, and halitosis of drinking, decoct soup are gargled.《Book on Chinese herbal medicine is stated》:Cold dispelling is wet, summer-heat and damp, hot and suffocating,
Damp and hot.Control diseases caused by external factors cold-evil, internal injury diet, or diet and hinder cold humidity hysteresis, mountainous evil air does not lie prostrate water and soil, and fever and chills make the diseases such as malaria.《Book on Chinese herbal medicine
New again》:Induce sweat and dissipate heresy, dampness removing removes wind, relieving heat and thirst.Control vomiting cholera, malaria, dysentery, sore scabies.Stalk:Larynx numbness, resolving sputum, cough-relieving can be controlled
Cough.
The invention has the beneficial effects as follows:
1) composition of the invention has the effect of uniqueness in terms of growth of animal, prevention and treatment IB is promoted.And because of medium-height grass
Medicine is safe and free of toxic and side effects, noresidue, prevention effect are good, low cost, does not affect to lay eggs;Compared with Western medicine, with the excellent of uniqueness
Gesture, its curative effect are better than antibiotics.
2) pharmacodynamic study result shows, the present invention composition its there is cough-relieving, antipyretic, antiinflammatory action.By observation
The therapeutic effect and clinical verification of artificial challenge IBV chickens are shown, after eating the preparation of the present invention, symptom mitigation, the state of mind
Take a turn for the better, feed intake increases, cough and asthma symptom disappears etc., only indivedual chickens occur getting rid of head, sneeze.The composition of the present invention effectively can delay
The solution diseased chicken clinical symptoms that cause of IBV, total effective rate reach more than 94%, cure rate and efficient be height compared with positive controls.
By the safety testing to target animals chicken, it is determined that this product is without toxicity.Its stable processing technique, quality controllable, it is suitable for
Big production is extracted, can large-scale application in the preventing and treating of IB.
Description of the drawings
Quantitative fluorescent PCR figures of the Fig. 1 for IBV;Wherein, amplification curve, solubility curve, real-time fluorescence are followed successively by from top to bottom
Quantitative PCR canonical plotting;
Fig. 2 sets up situation map for IB models;Wherein, a. is clinical, b. trachea and bronchus, c. tracheaes, d. hearts.
Fig. 3 lung tissues and tracheal tissue HE stained slice figures (40 times), wherein, A. lungs, B. bronchuses are 1. negative right
According to group;2. negative control group;3. oral liquid middle dose group of the present invention.
Specific embodiment
Embodiment 1:The preparation of powder
A kind of Chinese medicine composition of preventing and treating infectious bronchitis of chicken, is prepared from by following bulk drug:The 400g roots of large-flowered skullcap,
The 400g capsules of weeping forsythia, 400g cape jasmines, 400g Pogostemon cablin, 300g honeysuckles, 300g fritillaria thunbergiis, 200g radix bupleuri.Preparation technology:Weigh prescription
The root of large-flowered skullcap of amount, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and fritillaria thunbergii, the medicinal material in addition to the root of large-flowered skullcap first add prescription total weight
12 times amount water soak 1 hour, decoct 2 times, wherein the root of large-flowered skullcap after head boil medicine liquid boil after under, 2 hours every time, collecting decoction subtracted
Pressure is concentrated into thick paste, and vacuum drying is spray-dried, and is ground into fine powder, crosses 100 mesh sieves, adds 230g starch, mixes, and prepares
1000g powders.
Embodiment 2:The preparation of granule
A kind of Chinese medicine composition of preventing and treating infectious bronchitis of chicken, is prepared from by following bulk drug:The 400g roots of large-flowered skullcap,
The 400g capsules of weeping forsythia, 400g cape jasmines, 400g Pogostemon cablin, 200g honeysuckles, 200g fritillaria thunbergiis, 40g radix bupleuri.Preparation technology:Weigh prescription
The root of large-flowered skullcap of amount, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and fritillaria thunbergii, the medicinal material in addition to the root of large-flowered skullcap first add prescription total weight
12 times amount water soak 1 hour, decoct 2 times, wherein the root of large-flowered skullcap after head boil medicine liquid boil after under, 2 hours every time, collecting decoction subtracted
Pressure is concentrated into relative density 1.05~1.10 (50 DEG C), plus ethanol makes alcohol content up to 50%;10 DEG C arranged below 24 hours, leaching
Supernatant, reclaims ethanol to thick paste without alcohol taste, is evaporated to, and vacuum drying is spray-dried, and is ground into fine powder, crosses 100 mesh
Sieve, adds starch 200g and microcrystalline cellulose 400g, granulation to dry, whole grain, 1000g granules is obtained.
Embodiment 3:The preparation of granule
A kind of Chinese medicine composition of preventing and treating infectious bronchitis of chicken, is prepared from by following bulk drug:The 300g roots of large-flowered skullcap,
The 300g capsules of weeping forsythia, 300g cape jasmines, 300g Pogostemon cablin, 200g honeysuckles, 200g fritillaria thunbergiis, 100g radix bupleuri.Preparation technology:Weigh prescription
The root of large-flowered skullcap of amount, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and fritillaria thunbergii, the medicinal material in addition to the root of large-flowered skullcap first add prescription total weight
The water of 10 times of amounts soak 1 hour, decoct 3 times, wherein the root of large-flowered skullcap after head boil medicine liquid boil after under, 2 hours for the first time, second, the
Three times each 1 hour, collecting decoction, is evaporated to relative density 1.05~1.10 (50 DEG C), plus ethanol makes alcohol content up to 60%;
10 DEG C arranged below 48 hours, leaching supernatant, reclaims ethanol to thick paste without alcohol taste, is evaporated to, and vacuum drying is sprayed
Fine powder is dry, pulverize into, 100 mesh sieves are crossed, adds starch 300g and microcrystalline cellulose 360g, granulation to dry, whole grain, be obtained
1000g granules.
Embodiment 4:The preparation of capsule
A kind of Chinese medicine composition of preventing and treating infectious bronchitis of chicken, is prepared from by following bulk drug:The 120g roots of large-flowered skullcap,
The 120g capsules of weeping forsythia, 120g cape jasmines, 120g Pogostemon cablin, 60g honeysuckles, 60g fritillaria thunbergiis, 20g radix bupleuri.Preparation technology:Weigh recipe quantity
The root of large-flowered skullcap, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and fritillaria thunbergii, the medicinal material in addition to the root of large-flowered skullcap first adds prescription total weight 12
Times amount water soak 1 hour, decoct 2 times, wherein the root of large-flowered skullcap after head boil medicine liquid boil after under, 2 hours every time, collecting decoction, decompression
Relative density 1.05~1.10 (50 DEG C) is concentrated into, plus ethanol makes alcohol content up to 60%;10 DEG C arranged below 48 hours, in leaching
Clear liquid, reclaims ethanol to thick paste without alcohol taste, is evaporated to, and vacuum drying is spray-dried, and is ground into fine powder, crosses 100 mesh sieves,
Add starch 75g and microcrystalline cellulose 200g, granulation to dry, add a small amount of magnesium stearate, filling, 1000 capsules are obtained,
0.4g/ grains.
Embodiment 5:The preparation of powder
A kind of Chinese medicine composition of preventing and treating infectious bronchitis of chicken, is prepared from by following bulk drug:20 parts of roots of large-flowered skullcap, 20
Part capsule of weeping forsythia, 20 parts of cape jasmines, 15 parts of honeysuckles, 20 parts of Pogostemon cablin, 10 parts of radix bupleuri.Preparation technology:Weigh the root of large-flowered skullcap, the gold and silver of recipe quantity
Flower, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and fritillaria thunbergii, are ground into fine powder, cross 80-100 mesh sieves, mix, and packing is obtained powder.
Embodiment 6:The preparation of tablet
A kind of Chinese medicine composition of preventing and treating infectious bronchitis of chicken, is prepared from by following bulk drug:The 180g roots of large-flowered skullcap,
The 180g capsules of weeping forsythia, 180g cape jasmines, 180g Pogostemon cablin, 140g honeysuckles, 80g radix bupleuri, 140g fritillaria thunbergiis.Preparation technology:Weigh prescription
The root of large-flowered skullcap of amount, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and fritillaria thunbergii, the medicinal material in addition to the root of large-flowered skullcap first add prescription total weight
8 times amount water soak 1 hour, decoct 3 times, wherein the root of large-flowered skullcap after head boil medicine liquid boil after under, 2 hours every time, collecting decoction, decompression
Relative density 1.05~1.10 (50 DEG C) is concentrated into, plus ethanol makes alcohol content up to 70%;10 DEG C of 24-72 hours arranged below, filter
Supernatant being taken, ethanol being reclaimed to thick paste without alcohol taste, is evaporated to, vacuum drying is spray-dried, and is ground into fine powder, crosses 100
Mesh sieve, adds dextrin 90g and microcrystalline cellulose 200g, granulation to dry, adds magnesium stearate 10g, 1000, tablet is obtained, per
Piece 0.5g.
Embodiment 7:The preparation of oral liquid
A kind of Chinese medicine composition of preventing and treating infectious bronchitis of chicken, is prepared from by following bulk drug:The 150g roots of large-flowered skullcap,
The 150g capsules of weeping forsythia, 150g cape jasmines, 150g Pogostemon cablin, 100g honeysuckles, 100g fritillaria thunbergiis, 50g radix bupleuri.Preparation technology:Weigh prescription
The root of large-flowered skullcap of amount, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and fritillaria thunbergii, the medicinal material in addition to the root of large-flowered skullcap first add prescription total weight
8-12 times measure water immersion 0.5-3 hours, decoct 2-3 time, wherein the root of large-flowered skullcap after head boil medicine liquid boil after under, each 1-2 hours, conjunction
And decocting liquid, relative density 1.05~1.10 (50 DEG C) is evaporated to, plus ethanol makes alcohol content reach 50-70%;10 DEG C with decentralization
24-72 hours are put, leaching supernatant reclaims ethanol to without alcohol taste, adds 2g potassium sorbate preservatives, be diluted with water to
1000mL, filtration are filling, and sterilizing obtains final product oral liquid formulations.
Extract and study with refined selection process:
Consider that bird is administered feature, we preferably prepare oral liquid formulations, extract with reference to traditional Chinese medicine and Chinese veterinarian uses
Medicine experience, determining carries out water boiling and extraction active ingredient to the seven flavor medicine such as root of large-flowered skullcap material, using the preferred extracting parameter of orthogonal test.Mouthful
Liquid is taken because there is the requirement of clarity, and in order to ensure that oral liquid will not produce substantial amounts of precipitation during placement, in conjunction with
The property of each flavour of a drug in side, selects the extraction impurity-removing method of water extract-alcohol precipitation.Each factor such as considering cost and security, this work
Skill takes ethanol as removal of impurities solvent, carries out impurity removal process research to which.
1 test material
1.1 reagent:The seven flavor medicine material such as root of large-flowered skullcap (purchased from Anguo medicinal material market), scutelloside reference substance (the biological system of Chinese medicine
Institute, lot number 110715-200514 are determined in product examine).
1.2 instrument:High performance liquid chromatograph:1200 pump of Agilent, Agilent PDAD,
Agilent automatic sampler.
2 methods
2.1 decoctings boil orthogonal test
2.1.1 experimental design
Influence factor during according to water boiling and extraction, is examined with amount of water A, decocting time B, decoction number of times C as three
Factor is examined, three levels is taken respectively, using L9(34) table design orthogonal experiment, it is shown in Table 1.
1 water extraction quadrature factor water-glass of table
2.1.2 experimental technique
The 7 taste medicinal material such as the root of large-flowered skullcap, capsule of weeping forsythia 9 part is weighed respectively, by L9(34) orthogonal experimental method carries out technique investigation.Wherein yellow
Under a kind of reed mentioned in ancient books is after decocting liquid is boiled, with sample scutelloside amount contained by the root of large-flowered skullcap as index, preferred result (table 2, table 3).
Assay:Determine according to high performance liquid chromatography (the Chinese veterinary pharmacopoeia annex page 35 of version two in 2010).
Chromatographic condition is with system suitability with octadecylsilane chemically bonded silica as filler;- 0.4% phosphoric acid of methyl alcohol
(47: 53) it is mobile phase;Detection wavelength is 280nm.Theoretical cam curve is calculated with scutelloside peak and should be not less than 3000.
The preparation of reference substance solution takes scutelloside reference substance in right amount, accurately weighed, plus the dissolving of 50% methyl alcohol, makes every 1mL
Solution containing 50 μ g, obtains final product.
The preparation precision of need testing solution measures this product 1mL in 10mL measuring bottles, plus 50% methanol dilution is shaken to scale
Even, filter, take subsequent filtrate 2.5mL, be positioned in 10mL measuring bottles, plus 50% methyl alcohol is shaken up to scale, filter (0.45 μm of filter
Film), subsequent filtrate is used as need testing solution.
Determination method is accurate respectively to draw above-mentioned reference substance solution and each 10 μ L of need testing solution, injects liquid chromatograph, surveys
Fixed, obtain final product.
2.2 impurity removal process preferred
2.2.1 the selection of removal of impurities solvent
Medicinal material is weighed by 1000mL recipe quantities, carried out extracting, be concentrated into relative density 1.05 according to preferred extraction process by water
(50 DEG C), are divided into 10 parts, take 4 parts standby;Remaining 6 parts, 1 part of measure content of baicalin, another 5 parts add ethanol to make containing alcohol respectively
Amount carries out precipitation filtration up to 45%, 50%, 60%, 70%, 75%, and filtrate recycling ethanol is extremely without alcohol taste, plus potassium sorbate
0.2g, adds water to 100mL, shakes up, and filters, and determines content of baicalin, and observes liquid clarity after placing 1 week.
2.2.2 the investigation of reduced pressure concentration relative density
This preparation takes 60% ethanol as the solvent of purifying, and relative density of medicine liquid is less, and ethanol consumption is bigger, in liquid
Scutelloside to be dissolved in effect in ethanol solution better, but waste very much ethanol;Relative density of medicine liquid is bigger, consumes amount of alcohol and gets over
Little, therefore all dissolvings but active ingredient differs surely are investigated to the concentration relative density of liquid.
Standby liquid under above-mentioned 2.2.1 tests is taken, two parts are concentrated into relative density for 1.10 (50 DEG C of surveys), another two parts of concentrations
To the clear cream that relative density is 1.15 (50 DEG C of surveys), let cool, plus ethanol makes alcohol content up to 60%, stands overnight, filter, filter residue is done
Dry, to weigh, filtrate recycling ethanol respectively plus potassium sorbate 0.2g, adds water to 200mL, shakes up, filter, determine yellow to without alcohol taste
A kind of reed mentioned in ancient books glycosides content.
2.2.3 the investigation of standing time
The medicinal material of recipe quantity is weighed, totally 4 parts, according to above-mentioned preferred technique, plus 10 times of amount decoctings are boiled 3 times, and 2 is little for the first time
When (wherein the root of large-flowered skullcap after liquid boils under), second, each 1 hour for the third time, merge decoction liquor, filtration is evaporated to phase
To density 1.05~1.10 (50 DEG C), let cool, add ethanol, make alcohol content to 60%, respectively 10 DEG C 12 hours arranged below,
24 hours, 48 hours, 72 hours, leaching supernatant reclaimed ethanol to without alcohol taste, adds 2g potassium sorbates, add water regulation to
1000mL, filtration are filling, observe after placing one week.
2.3 process certification
The medicinal material of recipe quantity is weighed, totally three parts, respectively according to above-mentioned preferred technique, plus 10 times of amount decoctings boil 3 times (wherein
The root of large-flowered skullcap is added after after first time, liquid boils), 2 hours for the first time, second, each 1 hour for the third time, merge decoction liquor, filter,
Filtrate reduced in volume to relative density is the clear cream of 1.05~1.10 (50 DEG C of surveys), lets cool, adds ethanol, make alcohol content extremely
60%, to place 48 hours, leaching supernatant, ethanol is reclaimed to the regulation that without alcohol taste, adds water to 1000mL, filtration is filling, determines yellow
A kind of reed mentioned in ancient books glycosides content.
3 results
The orthogonal preferred parameter result of the test of 3.1 water extraction process
The preferred L9 (3 of 2 water extraction process of table4) Orthogonal experiment results
3 analysis of variance table of table
Analysis shows directly perceived, A2>A1>A3, B3>B2>B1, C3>C2>C1, i.e. best of breed is:A2B3C3, that is, add water 10 times and measure,
2 hours every time, decoct 3 times.Extreme difference R shows that impact order of each factor to water extraction process is C → A → B.
The result of variance analysis shows that factor C tool has a significant impact, therefore selection factor C affects maximum horizontal C3;A、
B does not make significant difference, i.e., A, B may be selected arbitrary level.Extraction can guarantee that again completely in view of energy saving, in conjunction with production warp
Test, factor A selects by-level A2, B selective extractions 2 hours, 1 hour, 1 hour.
Therefore this product optimum extraction process is:Above seven taste, adds water 10 times and measures, decoction 3 times, 2 hours for the first time, second,
Each 1 hour for the third time.
3.2 impurity removal process preferred results
3.2.1 alcohol precipitation solvent selection result is shown in Table 4.
4 alcohol precipitation concentration preferred result of table
Knowable to above experiment, when alcohol precipitation concentration reaches 45% and 75%, the content of scutelloside has greater loss, and alcohol precipitation is dense
Degree reaches 50%-70%, and content of baicalin change is less, illustrate relatively reasonable during alcohol precipitation concentration 50%-70%, because of 50% ethanol
Alcohol precipitation, supernatant and precipitation are in suspension, it is more difficult to filter, 70% ethanol alcohol precipitation is relatively costly, considers each factor,
60% ethanol is selected to carry out alcohol precipitation removal of impurities.
3.2.2 concentration densities selection result is shown in Table 5.
5 reduced pressure concentration relative density preferred result of table
From interpretation, when concentration of liquid medicine is to relative density 1.05,1.10, the basic phase of content of baicalin in liquid
When the precipitation capacity of generation is also substantially suitable, it is possible to determine that during relative density 1.05~1.10, alcohol precipitation substantially can be complete with removal of impurities;Phase
When being 1.15 to density, ethanol consumption is few, while the precipitation that liquid is produced is excessive, in liquid, content of baicalin is relatively low, and analysis may
It is part active ingredient to be removed in dedoping step.Consider each side factor, thus select concentration relative density be 1.05~
1.10(50℃).
3.2.3 after alcohol precipitation, standing time is investigated and the results are shown in Table 6.
The investigation result of standing time after 6 alcohol precipitation of table
As a result show, placing after alcohol precipitation 12 hours and filtering, final products have microprecipitation after placing one week, place 48 hours
Filter, final products are clarified after placing one week, meet oral liquid character requirement, therefore 10 DEG C of selection more than 48 hours arranged below is
Can.
3.3 process certification
Weigh the root of large-flowered skullcap, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and the fritillaria thunbergii of recipe quantity, the medicinal material in addition to the root of large-flowered skullcap
First plus 10 times of prescription total weight amount water immersion 0.5-3 hours, decoct 3 times, 2 hours for the first time, second, for the third time each 1
Hour, wherein the root of large-flowered skullcap after head boil medicine liquid boil after under, collecting decoction is evaporated to relative density 1.05~1.10 (50 DEG C),
Plus ethanol makes alcohol content up to 60%;10 DEG C arranged below 48 hours, leaching supernatant, reclaims ethanol to without alcohol taste, adds anti-corrosion
Agent, is diluted with water to 1000mL, filtration, filling, and sterilizing obtains final product oral liquid formulations.The results are shown in Table 7.
7 process certification result of table
As a result show, process certification result is basically identical with orthogonal preferred result, show preferred technological parameter repeat
Property is good.
Test example 1:Pharmacology test
Pharmacodynamics in terms of the present invention intends to cough-relieving, antipyretic, analgesia, anti-inflammatory are studied.
First, antitussive action research
Stimulated using the ammoniacal liquor that pharmacology is recorded and draw the method for coughing, simple to operate, economic and practical, therefore this test adopts excitant
Reagent ammoniacal liquor, makes ammoniacal liquor aerosol, stimulates mouse, causes mouse cough, so as to observe the antitussive action of the present composition.
1 test material
1.1 experimental animal
Using Test Animal Centre, Academy of Military Medical Sciences, P.L.A kunming mice (male, body weight 20~
22g, credit number:SCXK- (army) 2007-004).
1.2 medicines and reagent
Test medication:The oral liquid formulations of embodiment 5, this laboratory self-control, lot number:20140701,1mL liquid crude drugs contain
Amount 0.85g.It is administered orally.Positive control drug:Maxing Cough Syrup, Guangxi god of unusual strength Pharmacy stock Co., Ltd, lot number
140301, specification:100mL/ bottles.Ammoniacal liquor, Beijing Chemical Plant, lot number 110501 analyze pure.Sodium chloride injection, Sichuan Cologne
Medicine company limited company, lot number 20131025.
The multi-functional lured to cough to draw and breathes heavily instrument of 1.3 key instrument YLS-8A, Jinan Yi Yan development in science and technology Co., Ltd.
2 methods
2.1 screening ammoniacal liquor draw coughs administration time
9 mouse are taken, are divided into 3 groups, 3 per group, be respectively put into multi-functional lured to cough to draw and breathes heavily instrument, 20s, 40s is administered with ammoniacal liquor,
60s, observes the reaction of mouse, arrives cough number of times in the time of cough for the first time and 5min after record administration.Typically coughing is
Mouse contraction of abdominal muscle, while magnifying mouth, has cough.Mouse cough number of times is selected between 50~100, cough motion and sound is clear
Clear obvious, well-defined administration time group is defined as this test administration time.When showing administration 40s through result of the test, thorn
Sharp moderate, mouse can produce clear significantly cough, and action is obvious.
2.2 oral liquid cough-relieving tests of the present invention
Mouse 30 is taken, 5 groups are randomly divided into, 6 per group, first group of gavage gives the physiological saline of 1mL/kg bw, the
Two groups of gavages give Maxing Cough Syrup 1mL/kg bw, compare respectively as blank and positive drug, and other three groups give
Oral liquid of the present invention is basic, normal, high dosage group (0.75,1.5,3mL/kg bw).After administration 1h, mouse is put into multi-functional luring and is coughed
Draw and breathe heavily instrument, lured with ammoniacal liquor administration 40s and coughed, observe the reaction of mouse, the time after record administration to first time cough is (when hiding
Between) and the interior cough number of times of 5min.
2.3 statistical analysis
11.5 statistical softwares of application SPSS carry out statistical analysis, and Chi-square Test, variance analysis and Multiple range test carry out data
Analysis and the significance of difference compare, and result of the test is with average ± standard deviationRepresent, P<0.05 represents significant difference.
3 results
Result of the test is as shown in the table, and oral liquid of the present invention has good antitussive action, stops with positive control drug anesthesia apricot
Cough syrup effect slightly good, difference is not notable.With saline control group significant difference, cough latent time and 5min cough number of times
One times of difference, evident in efficacy.
The impact of the cough that the oral liquid of 8 present invention of table causes to ammoniacal liquor
Note:Same column compares, and adjacent letters represent significant difference (P<0.05), alternate letter representation difference extremely significantly (P<
0.01).Similarly hereinafter.
Determine that ammoniacal liquor draws first and cough required time, normal mouse is divided into five groups secondly, and gavage gives physiological saline respectively
(blank group), Maxing Cough Syrup (positive controls), (basic, normal, high) dosage oral liquid of the present invention, after 1 hour, ammoniacal liquor is lured
Cough, statistical result showed, positive drug group and investigational agent group latent time, cough number of times was few than blank group than blank group leader.
Show, the oral liquid energy of the present invention substantially reduces the mouse cough that ammoniacal liquor causes, illustrate that oral liquid of the present invention has preferable cough-relieving effect
Really.
2nd, refrigeration function research
This test fever model selects rat, exogenous heating agent to select 2,4-DNP, and the observation present invention is oral
The change to body temperature of liquid.
1 test material
1.1 experimental animal
Using the Wistar rats of Test Animal Centre, Academy of Military Medical Sciences, P.L.A, 30, male, body
Weigh 200~220g, credit number:SCXK- (army) 2007-004.
18~25 DEG C of airconditioning control room temperature, humidity 50%~70%, free water are ingested.
1.2 medicines and reagent
Test medication:5 oral liquid of the embodiment of the present invention, this laboratory self-control, product batch number:20140701,1mL liquids are given birth to
Medicine content 0.85g.It is administered orally.
Positive control drug:Aspirin tablet, Guangdong medicine company Industrial Co., Ltd. of health Botong, Chinese medicines quasi-word H44021139,
100mg/ pieces, face the used time with normal saline into 40mg/mL solution.
Sodium chloride injection, Kelun Pharm Ind Co., Ltd., Sichuan, lot number 20131025
2,4-DNP, Tianjin great Mao chemical reagent factories, lot number 20101202, analysis are pure, face used time physiology
Solution of the saline into 1.5mg/mL.
2 methods
Prepared by 2.1 2,4-DNP solution
Precision weighs 2,4-DNP 150mg, puts in 80mL physiological saline, is added dropwise over sodium hydroxide solution (dense
Degree:5mol/l), it is stirred continuously with magnetic stirring apparatus, is dissolved to 2,4-DNP just, is changed into glassy yellow clear solution,
Plus physiological water is settled to 100mL, obtain final product.
2.2 rat adaptability are raised
Male rat puts (23 ± 1 DEG C) adaptation 3d in laboratory environment, surveys 2 times/day of anus temperature, and continuous 3d first fits rat
The operation of anus temperature should be surveyed.Survey anus temperature 1 time, continuous survey 3 times were averaged and were designated as basal body temperature every 1 hour the test same day.Choose
Temperature changing, is grouped in 35.5~36.5 DEG C of rat at random less than 0.3 DEG C, mean body temperature.
2.3 solution heat tests
Choose and state rat 30, be divided into 5 groups, 6 per group.I.e. physiological saline group (equal-volume physiological saline), positive
Medicine control group (aspirin group 200mg/kg bw), the basic, normal, high dosage group (0.75,1.5,3mL/kg of oral liquid of the present invention
bw).After gastric infusion 1 hour, in mouse dorsal sc injection 2,4-DNP solution 2mL (concentration:1.5mg/mL), respectively
In hour each body temperature (DEG C) that surveys anus temperature once, count each minute point rat of record of 0.5, l, 1.5,2,4,6.With Temperature changing it is
Ordinate, minute point are abscissa, draw temperature curve.
2.4 statistical analysis
11.5 statistical softwares of application SPSS carry out statistical analysis, and Chi-square Test, variance analysis and Multiple range test carry out data
Analysis and the significance of difference compare, and result of the test is with average ± standard deviationRepresent, P<0.05 represents significant difference.
3 results
Test shows that the oral liquid of the present invention has certain refrigeration function.Later half little in injection 2,4-DNP
When, rat temperature rises, and reaches highest after about 1 hour, sustainable 3~4 hours.Pyrogenicity oral liquid height of the present invention after 1 hour
Dosage group body temperature is substantially less than nothing between physiological saline group and oral liquid low dosage of the present invention and middle dose group, with positive drug control group
Notable difference;2 hours and 4 hours, each test group body temperature of oral liquid of the present invention and positive drug control group body temperature were substantially less than and give birth to
Reason salt solution group;After 6 hours between each group body temperature without significant difference.As a result see the table below.
Impact (mean+SD, n=6) (DEG C) of the oral liquid of 9 present invention of table to 2,4-DNP pyrogenicity
By give five groups of rats, respectively gavage give physiological saline (blank group), aspirin (positive controls), (low,
Middle and high) dosage oral liquid of the present invention, after 1 hour, mouse dorsal sc injection 2,4-DNP solution 2mL, to different time
The body temperature of point carries out statistical analysis, as a result shows, after 1 hour, oral liquid of the present invention produces refrigeration function to pyrogenicity, and refrigeration function can
Continue 4 hours, indifference between each group after 6 hours.Show, oral liquid of the present invention has certain refrigeration function, but antipyretic
Effect is not so good as positive control drug aspirin.
3rd, antiinflammatory action research
This test selects following two inflammatory models, evaluates the antiinflammatory action of the oral liquid of the present invention, is that clinic is carried
For test basis.
1 test material
1.1 experimental animal
Using Test Animal Centre, Academy of Military Medical Sciences, P.L.A Wistar rats, 30, male, body weight
200~220g, credit number:SCXK- (army) 2007-004.
Using the kunming mice of Test Animal Centre, Academy of Military Medical Sciences, P.L.A, 30, male, body weight
18~22g, credit number:SCXK- (army) 2007-004.
20~25 DEG C of airconditioning control room temperature, humidity 50%~70%, free water are ingested.
1.2 medicines and reagent
Test medication:Oral liquid of the present invention, this laboratory self-control, product batch number:20140701,1mL liquid crude drug contents
0.85g.It is administered orally.
Positive control drug:Aspirin tablet, Guangdong medicine company Industrial Co., Ltd. of health Botong, Chinese medicines quasi-word H44021139,
100mg/ pieces, face the used time and pulverize, and with physiological saline solution, are configured to 40mg/mL solution.
2 methods
Impact of 2.1 oral liquids of the present invention to toes swelling caused by rat egg white
Wistar male rats 30 are chosen, stochastic averagina is divided into 5 groups, 6 per group.That is control group (equal-volume physiology salt
Water), positive drug control group (aspirin 200mg/kg bw), the basic, normal, high dosage group of oral liquid of the present invention (0.75,1.5,
3mL/kg bw), gastric infusion, 1 day 1 time, continuous 7 days.After last dose, with rat toes capacity measurer, measurement cause
A pawl volume after scorching front left.After last dose 1h, give rat left back pawl plantar subcutaneous injection fresh albumen, every 0.1mL is caused
Inflammation, respectively at cause inflammation after 0.5,1,2,4,6,8h, with rat toes capacity measurer measure cause inflammation after pawl volume so that scorching
Pawl volume difference is swelling in front and back.Calculate swelling and the inhibiting rate (%) of each group.
Sufficient pawl volume after toes swelling=cause inflammation causes the sufficient pawl volume before inflammation
Toes swelling inhibiting rate (%)=(control group toes swelling mean value-administration group toes swelling mean value)/
Control group toes swelling mean value × 100%
2.2 oral liquid paraxylene of the present invention cause the impact of mice auricle swelling
Kunming mice 30 is chosen, 5 groups are randomly divided into, 6 per group.That is control group (equal-volume physiological saline), positive drug
Control group (aspirin group 200mg/kg bw), the basic, normal, high dosage group (0.75,1.5,3mL/kg of oral liquid of the present invention
bw).Daily gastric infusion 1 time, continuous 7 days.After last dose l h, smear 0.02mL's to two sides before and after mouse right ear
Caused by dimethylbenzene xylene is scorching, and after l h, cervical dislocation puts to death mouse, and cuts ears along auricle baseline immediately, with the punching of a diameter of 6mm
Device, respectively in the same position punching of ears, uses electronic balance precise weighing, and ears weight difference is swelling;Swelling by each group
Degree carries out statistical analysis, calculates swelling rate average, standard deviation, inhibiting rate, compares the difference between each group.
Swelling (the mg)=left auricle weight of auris dextra piece weight
Swelling inhibiting rate (%)=(control group swelling mean value-administration group swelling mean value)/control group swelling
Degree mean value × 100%.
2.3 statistical analysis
11.5 statistical softwares of application SPSS carry out statistical analysis, and Chi-square Test, variance analysis and Multiple range test carry out data
Analysis and the significance of difference compare, and result of the test is with average ± standard deviationP<0.05 represents significant difference.
3 results
Impact of the oral liquid of 3.1 present invention to toes swelling caused by rat egg white
As known from Table 10, each dosage group of oral liquid of the present invention assumes inhibitory action, inhibitory action to the swelling of rat toes
With dosage into positive correlation.Result of the test shows, compared with control group, it is found that positive drug and each test group of oral liquid of the present invention have
The toes swelling effect for suppressing egg white to cause well, and oral liquid middle dose group of the present invention suppresses toes swelling effect and sun
Property medicine control group be close to.
Impact (mean+SD, n=6) of the oral liquid of 10 present invention of table to rat toes swelling caused by egg white
The oral liquid paraxylene of 3.2 present invention causes the impact of mice auricle swelling
As known from Table 11, oral liquid of the present invention can effectively suppress the mice auricle swelling that dimethylbenzene causes, inhibitory action with
Drug dose assumes positive correlation.Positive drug control group inhibitory action is between the middle and high dosage group of oral liquid of the present invention.
11 oral liquid paraxylene of the present invention of table causes the impact (mean+SD, n=6) of mice auricle swelling
Scorching method is caused using two kinds of egg white and dimethylbenzene.Egg white is caused in toes swelling experiment, to five groups of rat difference gavages
Physiological saline (blank group), aspirin (positive controls), (basic, normal, high) dosage oral liquid of the present invention is given, is even fed 7 days,
After last dose 1 hour, in rat left hind pawl plantar subcutaneous injection fresh albumen, sufficient pawl volume is determined respectively at different time,
Swelling is calculated, is as a result shown, a kind of reed mentioned in ancient books side oral liquid middle dose group is close to positive controls inhibiting rate.Caused by dimethylbenzene xylene ear swelling reality
In testing, to five groups of mouse, gavage gives physiological saline (blank group), aspirin (positive controls), (basic, normal, high) agent respectively
Amount oral liquid of the present invention, even hello 7 days, last dose smeared the dimethylbenzene of 0.02mL in two sides before and after mouse right ear after 1 hour,
Ear swelling degree is observed after 1 hour, is as a result shown, high dose group inhibiting rate is more obvious than positive controls high.Show of the invention oral
Liquid energy suppresses the rat toes swelling that egg white causes, and the mice ear that dimethylbenzene causes, with preferable antiinflammatory action.
The mechanism of our anti-inflammatory, it may be possible to produced by certain taste medicine, it is also possible to produced by the collaboration of square taste of traditional Chinese medicine.
Brief summary:The basic, normal, high dosage of oral liquid of the invention can reduce ammoniacal liquor and draw cough number of times, the suppression 2 for coughing mouse,
The rat temperature that 2, 4-dinitrophenol causes is raised, suppresses egg white to cause the toes swelling of rat and the swelling of auricle of mice caused by dimethylbenzene xylene
Swollen, with cough-relieving, antipyretic, antiinflammatory action.
Test example 2:Therapeutic effect to IB
This research, by observing therapeutic effect of the oral liquid of the present invention to artificial challenge IBV chickens, is further to develop and face
Bed application provides foundation.
1 test material
1.1 experimental animal
The 3 week old SPF Leghorns produced using Beijing Cimmeria Wei Tong animals used as test Technology Co., Ltd., male, 180
Only, licensing is numbered:SCXK (capital) 2009-0003.
1.2 test medications
Oral liquid of the present invention, this lab pilot plant product, product batch number:20140901,1mL liquid crude drug contents 0.85g.
It is administered orally.
Positive control drug:Close according to effect, similar comparable principle, from Maxingshigan oral liquid, beast in the favour of Luoyang
Medicine Co., Ltd, lot number:20140301, per 1mL equivalent to crude drug in whole 2.4g.Function with cure mainly:Clearing heat and detoxicating, a surname's lung cough-relieving,
Logical pharynx profit larynx.Cure mainly viral respiratory tract disease of fowl.Usage and dosage:Mixed drink, every 1 kg body weight, chicken, 1.5mL.
1.3 test strains
IBV:M41 strains, purchased from China Veterinery Drug Inspection Office, lot number:AV1511.
1.4 test reagents and instrument
Enzyme:RNase A, purchased from TIANGEN companies;Reverse transcriptase (M-MLV), Promega companies.
Carrier and bacterial strain:The PMD18T cloning vectors provided using Beijing Quanshijin Biotechnology Co., Ltd and DH5 α senses
Receive state cell.
Nucleic acid Marker:100bp DNA ladder, are provided by Beijing Hui Tian east Co., Ltd.
Kit:Green Realtime PCR Master Mix, Japanese TOYOBO Products;DNA fragmentation
Quick glue reclaim kit, vast Tyke biotechnology company;The little extraction reagent kit of high-purity plasmid, purchased from TIANGEN companies.
RNA extraction reagents:TRIzol LS Reagent:Promega Products.
Mx 3005P quantitative real time PCR Instruments:Stratagene companies.
1.5 test periods, place:This is tested on December 30,27 days to 2014 November in 2014 in Beijing Agricultural College's reality
Testing Animal House is carried out.
2 methods
2.1 rejuvenation virus EID50Measure
2.1.1 the rejuvenation of virus
1) pre-treatment is inoculated with:First with ovoscopy lamp in darkroom examination chicken embryo vigor, and with pencil drawing discharge chamber and the position of embryo
Put, and what a mark done at the position that will be inoculated with.When chick embryo development is normal, it is seen that the clearly chicken embryo of blood vessel and work, blood
Pipe and its Main Branches are obvious, and in cerise, chicken embryo can be with activity.The fixation of unfertilized and dead germ idiosome is motionless at one end, sees
Less than blood vessel, should pick out.
2) experimental implementation is required:All operations must rigorous aseptic.Laboratory, experiment super-clean bench all need to use ultraviolet in advance
Sterilization 20~30 minutes, experiment appliance autoclave sterilization sterilize.Inoculation operation is carried out in super-clean bench.
3) allantoic cavity inoculation
Chicken embryo is taken out, and (position of punching) is close at embryo position in chick embryo air sac and is carried out disinfection with iodine tincture cotton, alcohol swab, uses steel
Cone respectively wears an aperture after alcolhol burner flame calcination sterilization at air chamber top and air chamber lower edge are without blood vessel;IBV type strain M41
Press 1:10 dilutions, plus 4~6h of dual anti-rear 4 DEG C of effects, syringe inhales viral dilution liquid, subsequently by syringe needle from air chamber lower edge
Aperture is inserted perpendicularly into 0.5~1.0 centimeters (avoiding blood vessel), injects 0.2mL inoculums.And then paraffin sealing, juxtaposition is used to incubate
Uprightly it is incubated in ovum case, in 24h, died is discarded.Results allantoic fluid is carried out after 36~40h of incubation, is taken out and is put 4 DEG C of refrigerator overnights
In (general 10h) or -20 DEG C of refrigerators, 1.5h freezes to death chicken embryo, in order to avoid bleeding when harvesting.Collect allantoic fluid:Chicken embryo is taken out from refrigerator,
With the tincture of iodine or 75% alcohol disinfecting air chamber portion chorion, in the super-clean bench eggshell at tweezers strip off air chamber position, internal layer shell membrane is torn off
And CAM, embryo is gently pinned with tweezers, blood vessel is avoided with aseptic straw or sterile syringe and draws allantoic fluid, with
3000r/min is centrifuged 30min, takes supernatant, and packing after the chick embryo allantoic liquid of collection is merged is stored in -20 DEG C of refrigerators, in case
Next time passes on use.In 7 generation of above method blind passage, wait measure EID when passing to for 7 generation50.
2.1.2IBV EID50Determine
Allantoic fluid containing IBV-M41 is done 10 with PBS liquid-2~10-7Dilution, each dilution factor are inoculated with 10 age in days SPF chicken embryos
6 pieces, 0.1mL/ embryos are simultaneously provided with control inoculation physiological saline, to 20 ages in days, record dwarf embryo and death condition, use Reed-
Muench methods calculate its EID50, concrete formula is:
Distance than=(the infection percentage -50 higher than 50%)/(higher than 50% infection percentage-less than 50% sense
Dye percentage)
LgEID50=higher than 50% infection viral highest dilution logarithm+distance than the × coefficient of dilution logarithmic difference
The foundation of 2.2IBV models
According to multiple trial test result, 10 are adopted-3.30/ 0.1mL dosage is given and often supplies prelibation only for examination SPF chicken collunarium 0.2mL
Liquid.After attacking poison, chicken is isolated in closed environment and raises, daily (10 DEG C) stimulations of low temperature to 1~2 hour, and remaining time protects
Hold indoor temperature comfortable, relative humidity control observes clinical symptoms and pathology cut open inspection 70% ± 5%.
Treatment of 2.3 oral liquids of the present invention to IB
2.3.1 experimental animal packet and process
3 week old SPF male Leghorn 180, raises in Beijing Agricultural College's experimental animal room, is randomly divided into 6 groups, per group 30
Chicken.Negative control group, positive controls are individually raised in isolator, positive drug control group and oral liquid low dosage of the present invention
Group, oral liquid middle dose group of the present invention, oral liquid high dose group of the present invention are respectively at three layers of chicken coop raising of brooding.Do not contain in feed
Any medicine, free choice feeding and drinking-water.Adaptability attacks poison after feeding 4 days;After attacking poison, 48 hours all chick fall ill, and start to fill
Take drug treatment, successive administration 7 days.Test packet and process are shown in Table 12.
The test of table 12 packet and process
After 7 days treatment ends, each test group is scored according to evaluation of clinical curative effect standard, judge drug effect.
Test chicken is all cutd open and is killed, and wherein randomly selects each test group chicken 10, system cut open inspection, observe air bag transparency,
Whether tracheae and bronchus have oozes out and the cut open inspection pathological change such as lungs, to judge the curative effect of oral liquid of the present invention.
2.3.2IBV the foundation of carrying capacity assay method
2.3.2.1 Total RNAs extraction in tissue
100mg cryopreserved tissues are taken, 1mL TRIzol LS Reagent after grinding in liquid nitrogen, are added, after fully mixing, room
Warm 5min;0.2mL chloroforms are added according to every 1mL TRIzol LS Reagent, acutely vibration 15s, room temperature 3min, 4 DEG C
12000r/min is centrifuged 15min, draws upper strata aqueous phase;Addition 0.5mL isopropanols in upper strata aqueous phase, precipitation at room temperature 8min, 4 DEG C
12000r/min is centrifuged 10min;Tipping supernatant, add 75% ethanol 1mL rinsing after, abandon supernatant, room temperature air-dries 10min, with fit
Amount DEPC water dissolves are standby.
The sequence of the IBV strain M41 strains that has been delivered according to GenBank, designs pair of primers, amplification using Premier5.0
Fragment length is 227bp.Upstream primer 5 '-AAAACCAGTCCCAGATGCT-3 ';5 '-GAGGAAT of downstream primer
GAAATCCCAACG-3’.
2.3.2.2cDNA synthesis
The reverse transcription of the total serum IgE for being extracted as stated above, synthesizes cDNA.Press AMV ReverseTranscriptase
The synthesis for carrying out cDNA is described, sets up the reverse transcription system of 20 μ L, 2h at 42 DEG C, then 95 DEG C of denaturation 5min again, used as PCR moulds
Plate.20 μ L reverse transcription systems:
2.3.2.3 the preparation method (sequencing identification) of standard plasmid
The cDNA of acquisition is entered performing PCR amplification.PCR reaction systems are as follows:The each 5pmol of upstream and downstream primer, dNTPs
2 μ L of 5nmol, 10 × Buffer, cDNA 1 μ L, Taq archaeal dna polymerase 0.5U, plus ddH2The μ L of O to 20.PCR reaction conditions are:
95℃5min;94 DEG C of 30s, 55 DEG C of 30s, 72 DEG C of 30s, 30 circulations;72℃10min.PCR primer is carried out with 1.5% agarose
Gel electrophoresis is observed, recovery purifying purpose product fragment (using the quick glue reclaim kit of DNA fragmentation), builds IBV-PMD18T
The recombinant plasmid of standard, recombinant plasmid complete to be sequenced through Huada gene company, special with the IBV (M41 strains) of infection after comparison
Property fragment coincide.
The value that OD260 and OD280 is surveyed with nucleic acid concentration analyzer, is computed, and concentration is 1.1 × 1010Copies/ μ L, choosing
Plasmid with the ratio of OD260 and OD280 between 1.8~2.0, builds calibration curve.According to plasmid concentration is measured, as the following formula
Calculate copy number:
Copy number/μ L=6.02 × 1023(copied/mol) ÷ MW (g/mol) × concentration (g/ μ L)
Wherein, MW=purpose fragments size (bp) copies/ μ L × 660 (g/mol.bp)
According to the copy number for calculating, ten times of series of recombinant plasmid are diluted to containing 10 with TE buffer solutions10~
1038 gradients of copies/ μ L.DNA after with dilution repeats 2 as template, the template of each different dilution gradient
Hole, enters performing PCR amplification.As a result as shown in Figure 1.
2.3.3 MAIN OUTCOME MEASURES
2.3.3.1 production performance
From attack poison start calculate terminate to administration, totally 9 days.Record on-test is to terminating each group feed consumption respectively,
Calculate feed intake:Average daily gain (the g)=daily chicken number of total feed intake/∑.
10 chickens are randomly selected per group and is marked, when treatment starts with treatment end, by only weighing on an empty stomach, calculate average
Daily gain.Average daily gain (g)=(off-test body weight-on-test body weight)/number of days.
Feed-weight ratio:Feed-weight ratio=average daily gain/average daily gain
2.3.3.2 pathological change
Cut open inspection changes:Viewing test chicken nasal cavity, trachea-bronchial epithelial cell, lungs, air bag, liver, kidney etc. respectively organize organopathy
Reason change.
Histological observation:Lungs, kidney neutral formalin are fixed, FFPE, do histotomy, and HE is dyeed, tissues observed
Pathological change.
2.3.3.3 in tissue, viral nucleic acid carrying capacity is determined
Treatment 7 days after, every group of chicken takes 5, aseptic take trachea-bronchial epithelial cell, lungs and renal tissue, -80 DEG C protect
Standby, the to be set up assay method measure for organizing viral nucleic acid carrying capacity is deposited with 2.2.
2.3.3.4 curative effect determinate standard
Observation clinical symptoms, pathological change, judge curative effect in conjunction with the change of every Testing index.
Invalid:Spirit is highly depressed, drives to walk, and appetite is remarkably decreased, and gets rid of head, coughs, sneezes, tracheal sound.Cut open
Visible annulus trachealis hyperemia, bleeding is examined, has more serosity, Catarrhal and cheesy exudate;Visible bleeding in lung's bronchus,
Kidney uric acid mineralization.It is inefficiency that invalid chicken number accounts for the percentage of this group of chicken number.Effectively:Spiritual preferable, appetite is gradually extensive
Multiple, a small amount of chicken gets rid of head, coughs, sneezes.Cut open inspection observes a small amount of serosity of visible tracheae and Catarrhal mucus, and annulus trachealis is slight
Congested, rubescent, kidney lithate is deposited on a small quantity.The percentage that effectively chicken number accounts for this group of chicken number is efficient.Recovery from illness:Spirit,
Appetite, breathing recover normal, and cut open inspection trachea-bronchial epithelial cell, lungs, kidney etc. organize no abnormality seen.Recovery from illness chicken number accounts for this group of chicken
Several percentage is cure rate.Total effective rate:The percentage of this group chicken number is effectively accounted for recovery from illness chicken number summation.
2.4 statistical analysis
11.5 statistical softwares of application SPSS carry out statistical analysis, using Chi-square Test, variance analysis, Multiple range test, carry out
Data analysis and the significance of difference compare, result of the test average ± standard deviationTo represent, P<0.05 represents difference
Significantly.
3 results
3.1 half egg infectious amount (EID50) measurement result
Half egg infectious amount (EID is calculated according to Reed-Muench methods50):EID50Logarithm=higher than 50% virus dilute
Release logarithm=10 of logarithm+distance proportion × coefficient of dilution-6.30/0.1mL
Distance proportion=(higher than 50% chicken embryo death percentage -50%)/(higher than 50% chicken embryo death percentage-low
In 50% chicken embryo death percentage)=(8/14-50%)/(8/14-4/12)=0.3
LgEID50Difference between=distance proportion × dilution factor logarithm+it is higher than the dilution logarithm of 50% death rate
=0.3 × (- 1)+(- 6)=- 6.3
EID50=10-6.30/0.1mL.
13 IBV of table infects chicken embryo test process
Situation set up by 3.2IB models
It was found that test chicken involves rapidly full group in malicious 36h hinterlands supervention disease is attacked.Start within 3rd day symptom substantially, diseased chicken is main
Show as that spiritual depressed, uneasy or drowsiness, chilly is cold, flock together, loose random, the wing that hangs down are slow-witted vertical, get rid of head, viscous tears, sneeze, breathe heavily
Cough, obvious tracheal sound occur, evening symptom is particularly evident.Cut open inspection finds diseased chicken nasal meatus, trachea-bronchial epithelial cell mucous membrane swelling,
Chamber is contained within slurries, mucus or the cheesy exudate of varying number;Bronchus and tracheae intersection have a bleeding, and the ring of tracheae
Shape part also assumes bleeding;Air bag is muddy;Heart born of the same parents' hydrops is serious.Above symptom meets chicken and passes the clinical symptoms that props up.Knot
Fruit is as shown in Figure 2.As a result show, IB models are successfully established, for prelibation liquid with 10-3.30/ 0.1mL concentration is to often only for examination SPF chickens
Collunarium 0.2mL, is the infective dose of this test model.
Curative effect of 3.3 oral liquids of the present invention to IB morbidity chickens
3.3.1 the impact to production performance
14 result of the test of table shows, compared with positive controls, negative control group, positive drug control group and the present invention are oral
The average daily gain of the test chicken of the high, medium and low dosage group of liquid dramatically increases (P<0.05);Show from feed-weight ratio, positive right
Increase according to group feed-weight ratio, and positive drug control group, oral liquid senior middle school low dose group feed-weight ratio of the present invention are lower slightly compared with control group.
Impact of 14 oral liquid of the present invention of table to IB chicken production performances (N=10)
3.3.2 histological change
Positive controls lung group can be caused after breathing channel type M41 strain infection SPF chickens are understood by histopathologic slide's figure of Fig. 2
Knit, alluvial around alveolar has substantial amounts of red blood cell, has red blood cell it can in addition contain see in bronchus tube chamber, and the present invention is oral
Liquid treatment group can be clearly apparent lobulus pulmonis and organizations at different levels' structure, rarely seen a small amount of red blood cell.Bronchial wall hyperemia, oedema and lymph
Cell, plasmocyte infiltrating;Tracheal mucosa epithelial cell denaturation, partial necrosis come off, and in mucosal epithelium, goblet cell increases.Through
After oral liquid treatment of the present invention, above-mentioned pathology has clear improvement.
In summary, oral liquid of the present invention is damaged to chicken lung, tracheal tissue caused by breathing channel type IBV
Wound has obvious therapeutic effect.
3.3.3 the measure of viral nucleic acid carrying capacity in organizing after treating
As shown in Table 15, IBV nucleic acid loads generation significant change in organizing after treatment in 7 days, positive drug control group, this
The high, medium and low dosage group viral nucleic acid carrying capacity of invention oral liquid significantly reduces (P<0.05), and positive controls tissue in IBV disease
Malicious nucleic acid load highest.
Table 15IBV infected chickens treatment after IBV nucleic acid load results (10x,N=5) (logcopy)
3.3.4 impact of the oral liquid of the present invention to IB chicken clinical symptoms
After adaptability is raised, with the chick embryo allantoic liquid of IBV infection, through collunarium point ocular infections chick, part after 24 hours
Chicken falls ill, and after 48 hours, all chick fall ill;It is mainly shown as that spirit is highly depressed, stands transfixed to the ground, cough stream in disorder by plumage
The symptoms such as tears, breath sound, expiratory dyspnea, anorexia, feed intake minimizing.After oral liquid of the present invention and positive drug treatment, disease
Shape mitigates, and the state of mind takes a turn for the better, and feed intake increases, and cough and asthma symptom disappears etc., and only indivedual chickens occur getting rid of head, sneeze.
Impact of the oral liquid of 16 present invention of table to IB chicken curative effects
Result of the test shows, compared with positive controls, the basic, normal, high dosage group of oral liquid of the present invention effectively can be alleviated
The diseased chicken clinical symptoms that IBV causes, wherein with high dose group effect preferably, total effective rate reaches 83.33%, middle dose group effect
Take second place, between high, medium and low dosage group, total effective rate is in dose-effect dependence.
Test example 3:Clinical verification
Experimental animal:According to epidemiology, clinical symptoms, pathology cut open inspection, identification and isolation of pathogen, through comprehensive diagnos for passing
The diseased chicken of metachromia bronchitis 400, is randomly divided into 4 groups, 100 per group, respectively 4 groups of embodiment, 5 groups of embodiment, red mould
Plain control group and blank control group.
Trial drug:Chinese medicine composition prepared by embodiment 4-5 is added in feed according to 0.8% weight ratio, is mixed thoroughly,
Diseased chicken is fed respectively.Every kilogram of basal diet of erythromycin control group fed adds the feed of 150,000 IU erythromycin.Blank control group
The edible feed for being not added with any medicine.It is used in conjunction 7 days.
Treatment standard:1) fully recover:Coughed with diseased chicken, sneeze, chick flow nose liquid, chicken egg productivity of laying eggs is reduced, and respiratory tract glues
In serosity, it is standard that the symptom such as catarrhal inflammation disappears to film;2) effectively:Coughed with diseased chicken, sneeze, chick flow nose liquid, lay eggs
Chicken egg productivity is reduced, and in serosity, the remission such as catarrhal inflammation is standard to respiratory mucosa;3) invalid:Symptom does not improve,
Spirit is highly depressed, drives to walk, and appetite is remarkably decreased, and gets rid of head, coughs, sneezes, tracheal sound.The visible annulus trachealis of cut open inspection fills
Blood, bleeding, have more serosity, Catarrhal and cheesy exudate;Visible bleeding in lung's bronchus, kidney lithate are sunk
Product.Total effective rate:The percentage of this group chicken number is effectively accounted for recovery from illness chicken number summation.
Clinical treatment outcome of the oral liquid of 17 present invention of table to IB chickens
After the preparation of the present invention and positive drug treatment, symptom mitigation, the state of mind take a turn for the better, and feed intake increases, cough and asthma symptom
Disappear etc., only indivedual chickens occur getting rid of head, sneeze.As a result show, the composition of the present invention can effectively alleviate the disease that IBV causes
Chicken clinical symptoms, total effective rate reach more than 94%, cure rate and efficient be height compared with positive controls.
Claims (10)
1. a kind of preventing and treating infectious bronchitis of chicken Chinese medicine composition, it is characterised in that the Chinese medicine composition is by following weight portion
Several bulk drugs is prepared from:100-200 part roots of large-flowered skullcap, 100-200 part capsules of weeping forsythia, 100-200 part cape jasmines, 50-150 part gold and silver
Flower, 100-200 part Pogostemon cablin, 10-100 part radix bupleuri, 50-150 part fritillaria thunbergiis.
2. according to claim 1 preventing and treating infectious bronchitis of chicken Chinese medicine composition, it is characterised in which is by following
The bulk drug of weight proportion is prepared from:120-180 part roots of large-flowered skullcap, 120-180 part capsules of weeping forsythia, 120-180 part cape jasmines, 60-140 parts
Honeysuckle, 120-180 part Pogostemon cablin, 20-80 part radix bupleuri, 60-140 part fritillaria thunbergiis.
3. according to claim 1 preventing and treating infectious bronchitis of chicken Chinese medicine composition, it is characterised in which is by following
The bulk drug of weight proportion is prepared from:140-160 part roots of large-flowered skullcap, 140-160 part capsules of weeping forsythia, 140-160 part cape jasmines, 80-120 parts
Honeysuckle, 140-160 part Pogostemon cablin, 40-60 part radix bupleuri, 80-120 part fritillaria thunbergiis.
4. according to the Chinese medicine composition of the arbitrary described preventing and treating infectious bronchitis of chicken of claim 1-3, it is characterised in that:Institute
Chinese medicine composition is stated for powder, capsule, oral liquid, granule, parenteral solution or tablet.
5. the preparation method of the Chinese medicine composition of preventing and treating infectious bronchitis of chicken according to claim 4, its feature exists
As follows in step:The root of large-flowered skullcap, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and the fritillaria thunbergii of recipe quantity is weighed, fine powder is ground into,
80-100 mesh sieves are crossed, is mixed, packing is obtained powder.
6. a kind of as claimed in claim 4 preventing and treating infectious bronchitis of chicken Chinese medicine composition preparation method, its feature
It is that step is as follows:The root of large-flowered skullcap, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and the fritillaria thunbergii of recipe quantity is weighed, in addition to the root of large-flowered skullcap
Medicinal material first adds the water immersion 0.5-3 hours that 8-12 times of prescription total weight measures, and decocts 2-3 time, and wherein the root of large-flowered skullcap treats that head liquid of boiling medicine is boiled
Under after boiling, each 1-2 hours, collecting decoction are evaporated to thick paste, and vacuum drying is spray-dried, and is ground into fine powder, mistake
100 mesh sieves, add appropriate amount of auxiliary materials, mix, prepare capsule, granule or tablet.
7. a kind of as claimed in claim 4 preventing and treating infectious bronchitis of chicken Chinese medicine composition preparation method, its feature
It is that step is as follows:The root of large-flowered skullcap, honeysuckle, the capsule of weeping forsythia, cape jasmine, Pogostemon cablin, radix bupleuri and the fritillaria thunbergii of recipe quantity is weighed, in addition to the root of large-flowered skullcap
Medicinal material first adds the water immersion 0.5-3 hours that 8-12 times of prescription total weight measures, and decocts 2-3 time, and wherein the root of large-flowered skullcap treats that head liquid of boiling medicine is boiled
Under after boiling, each 1-2 hours, collecting decoction are evaporated to relative density 1.05~1.10 (50 DEG C), plus ethanol makes alcohol content
Reach 50-70%;10 DEG C of 24-72 hours arranged below, leaching supernatant reclaim ethanol to without alcohol taste, add preservative, add water dilute
Release to oral liquid 0.6~1.0g containing primary crude drug of every 1mL, filtration is filling, and sterilizing obtains final product oral liquid formulations.
8. according to claim 6 or 7 preventing and treating scouring Chinese medicine composition preparation method, it is characterised in that:Carry
Taking technique parameter is:Add water 10 times and measure, decoct 3 times, for the first time 2 hours, second, each 1 hour for the third time.
9. according to claim 7 preventing and treating scouring Chinese medicine composition preparation method, it is characterised in that:Remove general labourer
Skill parameter is:Extract is concentrated into relative density 1.05~1.10 (50 DEG C), plus ethanol makes alcohol content up to 60%.
10. a kind of Chinese medicine composition according to claim 1 is used for treating in preparation or prevents infectious bronchitis of chicken
In application.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201611017765.1A CN106421322A (en) | 2016-11-15 | 2016-11-15 | Traditional Chinese medicine composition capable of preventing and treating infectious bronchitis of chickens and preparation method of traditional Chinese medicine composition |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201611017765.1A CN106421322A (en) | 2016-11-15 | 2016-11-15 | Traditional Chinese medicine composition capable of preventing and treating infectious bronchitis of chickens and preparation method of traditional Chinese medicine composition |
Publications (1)
Publication Number | Publication Date |
---|---|
CN106421322A true CN106421322A (en) | 2017-02-22 |
Family
ID=58220169
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201611017765.1A Pending CN106421322A (en) | 2016-11-15 | 2016-11-15 | Traditional Chinese medicine composition capable of preventing and treating infectious bronchitis of chickens and preparation method of traditional Chinese medicine composition |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN106421322A (en) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102657724A (en) * | 2012-05-23 | 2012-09-12 | 天津生机集团股份有限公司 | Traditional Chinese medicine oral liquid for preventing and treating chicken infectious bronchitis and preparation method thereof |
CN104208393A (en) * | 2013-09-30 | 2014-12-17 | 郑州后羿制药有限公司 | Traditional Chinese medicine composition for treating chicken infectious bronchitis and preparation method thereof |
CN104706993A (en) * | 2013-12-11 | 2015-06-17 | 天津嘉创生物科技有限公司 | Traditional Chinese medicine oral liquid with functions of heat clearing, detoxification, pungent cooling and exterior syndrome relieving, and preparation method thereof |
-
2016
- 2016-11-15 CN CN201611017765.1A patent/CN106421322A/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102657724A (en) * | 2012-05-23 | 2012-09-12 | 天津生机集团股份有限公司 | Traditional Chinese medicine oral liquid for preventing and treating chicken infectious bronchitis and preparation method thereof |
CN104208393A (en) * | 2013-09-30 | 2014-12-17 | 郑州后羿制药有限公司 | Traditional Chinese medicine composition for treating chicken infectious bronchitis and preparation method thereof |
CN104706993A (en) * | 2013-12-11 | 2015-06-17 | 天津嘉创生物科技有限公司 | Traditional Chinese medicine oral liquid with functions of heat clearing, detoxification, pungent cooling and exterior syndrome relieving, and preparation method thereof |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN101007067A (en) | Antivirus medicine for birds and its preparation method | |
CN104436142B (en) | A kind of Chinese medicine composition for treating pig epidemic diarrhea | |
CN104027442B (en) | A kind of Chinese medicine preparation, preparation method and the method for use Chinese medicine preparation cultivation broiler chicken used in green no drug residue broiler breeding | |
CN102895326B (en) | Traditional Chinese medicine composition for treating infantile common cold and preparation method for traditional Chinese medicine composition | |
CN104887890A (en) | Medicine composition for treating white scour of chicken and preparation method thereof | |
CN103494927B (en) | Traditional Chinese medicine composition for treating chicken respiratory disease and preparation method thereof | |
CN102696910A (en) | Feed composition and traditional Chinese medicine for preventing and treating avian infectious laryngotracheitis | |
CN105213993A (en) | One improves cachectic Chinese medicine composition of intestinal cancer and preparation method thereof | |
CN102552657B (en) | Chinese medicinal composition used for treating Newcastle disease of chicken | |
CN102188494A (en) | Medicament for treating high fever of pigs with pure Chinese medicine | |
CN106177900B (en) | A kind of gel of anti-human papilloma virus (anti-HPV) and preparation method thereof | |
CN106421322A (en) | Traditional Chinese medicine composition capable of preventing and treating infectious bronchitis of chickens and preparation method of traditional Chinese medicine composition | |
CN103721147B (en) | For treating the Chinese medicine composition of chicken coccidiasis | |
CN103623192B (en) | Medicine being used for the treatment of pig blue-ear disease and preparation method thereof | |
CN104784657B (en) | A kind of application of herbal medicine compound preparation in treating chicken colibacillosis | |
CN113101331B (en) | Thyme herb tea and preparation method and application thereof | |
CN103041172B (en) | Traditional Chinese medicine composition for curing chronic nasosinusitis and preparation method thereof | |
CN105031147B (en) | A kind of Chinese medicine composition and its preparation method and application | |
CN105770265A (en) | Medicinal preparation for treating esophageal carcinoma and application thereof | |
CN105853740A (en) | Anesthetic for stomachoscopy | |
CN106038765A (en) | Application of compound Rabdosia rubescens preparation to chemical prevention of esophagus cancer | |
CN105663899A (en) | Traditional Chinese medicine preparation for treating pulmonary abscess | |
CN106265658B (en) | Application of the flat shellfish ketone in preparation prevention and/or treatment herpes virus hominis's infection medicine | |
CN103705820B (en) | Medicine being used for the treatment of yellow scour of piglet and preparation method thereof | |
CN104224929A (en) | Traditional Chinese medicine composition for treating common cold and upper respiratory tract infection as well as preparation method and application thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
RJ01 | Rejection of invention patent application after publication | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170222 |