The specific embodiment
Embodiment 1
The Chinese medicine of preparation treatment tonsillitis
1, gets the raw materials ready: get 490 parts of Fructus Arctiis; 320 parts of Lasiosphaera Seu Calvatia; 490 parts of Herba Schizonepetae; 490 parts of Bombyx Batryticatus; 160 parts of 490 parts of Periostracum Cicadaes and Rhizoma Cimicifugaes.Fructus Arctii wherein: remove impurity, cleaning, drying; Lasiosphaera Seu Calvatia: remove impurity, be cut into small pieces; Herba Schizonepetae: remove impurity, the spray clear water is cleaned, and runs through, and cutting is dried; Bombyx Batryticatus: eluriate after drying, remove impurity; Periostracum Cicadae: remove impurity, clean, dry; Rhizoma Cimicifugae: remove impurity, slightly bubble is cleaned, and runs through, and cuts sheet, drying.
2, Fructus Arctii is extracted: Fructus Arctii, add 6 times of amount 60% alcohol reflux secondaries, each 2 hours, merge extractive liquid, filtered, filtrate recycling ethanol, the thick paste that is condensed into is standby;
3, Herba Schizonepetae is extracted: get Herba Schizonepetae and add 8 times of water gagings, distillating extracting oil 1.5 hours, collect volatile oil, volatile oil is dissolved in ethanol (volatile oil-ethanol 1: 3), join in 40 ℃ the full Heshui solution of beta-schardinger dextrin-(volatile oil-beta cyclodextrin-water ratio is 1: 8: 80), 40 ℃ of constant temperature stirred 2 hours, cold preservation 24 hours filters 50 ℃ of dryings, sieve, standby; Aqueous solution after distillation device is in addition collected standby.
4, Lasiosphaera Seu Calvatia, Bombyx Batryticatus, Periostracum Cicadae and Rhizoma Cimicifugae are extracted: with adding 10 times of water gagings in Lasiosphaera Seu Calvatia, Bombyx Batryticatus, Periostracum Cicadae and the Rhizoma Cimicifugae, decoct secondary, each 1 hour, collecting decoction filtered, and filtrate and above-mentioned aqueous solution merge, and are concentrated into into thick paste.Here, the medicinal residues after also Herba Schizonepetae can being extracted add extraction in the lump in Lasiosphaera Seu Calvatia, Bombyx Batryticatus, Periostracum Cicadae and the Rhizoma Cimicifugae, thereby Herba Schizonepetae is further extracted.
5, mix: the thick paste in the thick paste and 2 that obtains in 4 is merged, and drying under reduced pressure (0.08Mpa, 80 ℃) is ground into fine powder, adding Herba Schizonepetae volatile oil clathrate and right amount of auxiliary materials, and with ethanol system granule, 50 ℃ of dryings, that is, the adjuvant here is dextrin and sucrose.
Embodiment 2
The Chinese medicine of preparation treatment tonsillitis
1, gets the raw materials ready: get 510 parts of Fructus Arctiis; 340 parts of Lasiosphaera Seu Calvatia; 510 parts of Herba Schizonepetae; 510 parts of Bombyx Batryticatus; 170 parts of 510 parts of Periostracum Cicadaes and Rhizoma Cimicifugaes.Fructus Arctii wherein: remove impurity, cleaning, drying; Lasiosphaera Seu Calvatia: remove impurity, be cut into small pieces; Herba Schizonepetae: remove impurity, the spray clear water is cleaned, and runs through, and cutting is dried; Bombyx Batryticatus: eluriate after drying, remove impurity; Periostracum Cicadae: remove impurity, clean, dry; Rhizoma Cimicifugae: remove impurity, slightly bubble is cleaned, and runs through, and cuts sheet, drying.
2, Fructus Arctii is extracted: Fructus Arctii, add 6 times of amount 60% alcohol reflux secondaries, each 2 hours, merge extractive liquid, filtered, filtrate recycling ethanol, it is standby to be condensed into thick paste;
3, Herba Schizonepetae is extracted: get Herba Schizonepetae and add 8 times of water gagings, distillating extracting oil 1.5 hours, collect volatile oil, volatile oil is dissolved in ethanol (volatile oil-ethanol 1: 3), join in 40 ℃ the full Heshui solution of beta-schardinger dextrin-(volatile oil-beta cyclodextrin-water ratio is 1: 8: 80), 40 ℃ of constant temperature stirred 2 hours, cold preservation 24 hours filters 50 ℃ of dryings, sieve, standby; Aqueous solution after distillation device is in addition collected standby.
4, Lasiosphaera Seu Calvatia, Bombyx Batryticatus, Periostracum Cicadae and Rhizoma Cimicifugae are extracted: with adding 10 times of water gagings in Lasiosphaera Seu Calvatia, Bombyx Batryticatus, Periostracum Cicadae and the Rhizoma Cimicifugae, decoct secondary, each 1 hour, collecting decoction filtered, and filtrate and above-mentioned aqueous solution merge, and are condensed into thick paste.Here, the medicinal residues after also Herba Schizonepetae can being extracted add extraction in the lump in Lasiosphaera Seu Calvatia, Bombyx Batryticatus, Periostracum Cicadae and the Rhizoma Cimicifugae, thereby Herba Schizonepetae is further extracted.
5, mix: the thick paste in the thick paste and 2 that obtains in 4 is merged, and drying under reduced pressure (0.08Mpa, 80 ℃) is ground into fine powder, adds the Herba Schizonepetae volatile oil clathrate, reaches right amount of auxiliary materials, and with ethanol system granule, 50 ℃ are drying to obtain, and the adjuvant here is dextrin and sucrose.
Most preferred embodiment
The Chinese medicine of preparation treatment tonsillitis
1, gets the raw materials ready: get 500 parts of Fructus Arctiis; 333 parts of Lasiosphaera Seu Calvatia; 500 parts of Herba Schizonepetae; 500 parts of Bombyx Batryticatus; 167 parts of 500 parts of Periostracum Cicadaes and Rhizoma Cimicifugaes.Fructus Arctii wherein: remove impurity, cleaning, drying; Lasiosphaera Seu Calvatia: remove impurity, be cut into small pieces; Herba Schizonepetae: remove impurity, the spray clear water is cleaned, and runs through, and cutting is dried; Bombyx Batryticatus: eluriate after drying, remove impurity; Periostracum Cicadae: remove impurity, clean, dry; Rhizoma Cimicifugae: remove impurity, slightly bubble is cleaned, and runs through, and cuts sheet, drying.
2, Fructus Arctii is extracted: Fructus Arctii, add 6 times of amount 60% alcohol reflux secondaries, each 2 hours, merge extractive liquid, filtered, filtrate recycling ethanol, it is standby to be condensed into thick paste;
3, Herba Schizonepetae is extracted: get Herba Schizonepetae and add 8 times of water gagings, distillating extracting oil 1.5 hours, collect volatile oil, volatile oil is dissolved in ethanol (volatile oil-ethanol 1: 3), join in 40 ℃ the full Heshui solution of beta-schardinger dextrin-(volatile oil-beta cyclodextrin-water ratio is 1: 8: 80), 40 ℃ of constant temperature stirred 2 hours, cold preservation 24 hours filters 50 ℃ of dryings, sieve, standby; Aqueous solution after distillation device is in addition collected standby.
4, Lasiosphaera Seu Calvatia, Bombyx Batryticatus, Periostracum Cicadae and Rhizoma Cimicifugae are extracted: with adding 10 times of water gagings in Lasiosphaera Seu Calvatia, Bombyx Batryticatus, Periostracum Cicadae and the Rhizoma Cimicifugae, decoct secondary, each 1 hour, collecting decoction filtered, and filtrate and above-mentioned aqueous solution merge, and are condensed into thick paste.Here, the medicinal residues after also Herba Schizonepetae can being extracted add extraction in the lump in Lasiosphaera Seu Calvatia, Bombyx Batryticatus, Periostracum Cicadae and the Rhizoma Cimicifugae, thereby Herba Schizonepetae is further extracted.
5, mix: the thick paste in the thick paste and 2 that obtains in 4 is merged, and drying under reduced pressure (0.08Mpa, 80 ℃) is ground into fine powder, adds the Herba Schizonepetae volatile oil clathrate, reaches right amount of auxiliary materials, and with ethanol system granule, 50 ℃ are drying to obtain, and the adjuvant here is dextrin and sucrose.
In the present invention, medicine can be made granule, can tabletting or dress up capsule.
Pharmacodynamics test of the present invention:
Experiment material
1.1 be subjected to the reagent thing
The granular medicament that prescription of the present invention and technology are made, name are called the granule excipient contrast that disappears of the vigorous tonsillitis of burdock: excipient is made up of dextrin, Icing Sugar, cyclamate, is made into 65% concentration, presses 10ml/kg filling stomach.
Positive control drug (one): aspirin (enteric coatel tablets), every contains aspirin 25mg, clinical adult's every day oral 50mg, selecting mice dosage is 0.3g/kg, rat dosage is 0.2g/kg.
Positive control drug (two): the amoxicillin sheet, selecting mice dosage is 0.52g/kg, is equivalent to 2 times of dose,equivalents.
Positive control drug (three): guanidine quadrhydrochloride (Guanidine HCL), face the time spent to be mixed with desired concn with the RPMI1640 cell culture fluid.Be used for the extracorporeal antivirus effect test.
Positive control drug (four): LINGDANCAO JIAONANG, selecting mice dosage is 0.7g/kg, is equivalent to 2 times of dose,equivalents.
Positive control drug (five): cloperastine tablet, every contains the 10mg cloperastine hydrochloride, lot number: 20040101.Selecting mice dosage is 0.5g/kg, is equivalent to 2 times of dose,equivalents.
1.2 experimental animal
Closed colony, outbreeding system, Wistar rat, body weight 120~150 grams, male and female half and half; Kunming kind white mice, body weight 18~20 grams, male and female half and half.
1.2. animal feeding condition:
Animal housing is a regular grade, room temperature: 25 ℃, and relative temperature: 40%.Ventilate, illumination is good, adopts standard feed to feed.Animal is that female, the male separately same sex is mixed the cage raising, 5 of every cage animals, and every morning timing administration and the feedwater of quantitative feeding 1 time, and measure its preceding 1 day surplus.
1.3 test virus
Coxsackie B 3 viruses (Coxsackie B3 virus), the Nancy strain
Adenovirus type VII (Ad
7)
Influenza virus (FM1)
1.4 strain and bacterium number:
National standard strain: staphylococcus aureus 26003
Colon bacillus 31001
Local strain: klepsiella pneumoniae 13883
Streptococcus faecalis ATCC 29212
Fu Shi will Hayes ATCC 12022
1.5 cell: people's amniotic membrane (FL) cell.Madin-Darby canine kidney(cell line) (MDCK) (MDCK).
1.6 reagent: the RPMI-1640 cell culture fluid; Newborn calf serum; Trypsin; (MTT, 3-4,5-dimethylthiazo are (2-yl)-2,5-diphenylte-trazoliumdromide) for tetrazolium bromide.
2. dosage is selected
Because median lethal dose(LD 50) LD
50Fail to measure, so select the animal-use drug amount with reference to clinical medicine dose and maximum dosage measured value.Clinical per day for adults clothes 30g crude drug, be equivalent to 0.43g crude drug/kg, pressing human body and animal body surface area equivalent dose ratio calculates, selecting 3 kinds of dosage of mice is 15.6g, 7.8g, 3.9g crude drug/kg, 3 kinds of dosage of rat are 10.8g, 5.4g, 2.7g crude drug/kg (being respectively 4 times, 2 times and dose,equivalent), the equal gastric infusion of animal (ig).Route of administration is with clinical consistent.
3. test method and result
3.1, antiinflammatory test
3.1.1. the vigorous tonsillitis of burdock disappears to the influence of Ovum Gallus domesticus album rat paw edema
[2]
Get 50 of rats, male and female half and half are divided into 6 groups at random, 10 of every treated animals,, the 1st, 2,3 group is the vigorous tonsillitis of the burdock high, medium and low dosage group that disappears, dosage is respectively 10.8,5.4,2.7g crude drug/kg, (being equivalent to 4 times, 2 times, dose,equivalent respectively), the 4th group of positive medicine aspirin (administration on the same day 1 time), dosage 0.2g/kg,, the 5th group is matched group, each organizes ig administration every day according to dosage 1 time, successive administration 3 days, matched group gives excipient by the administration high dose.Irritate stomach (ig) administration.Measure the right back ankle of rat joint girth as causing scorching preceding girth and record with self-control arrowband chi during test, behind last administration 30min, respectively only at the right back sufficient subcutaneous injection fresh albumen 0.1ml/ of rat.Survey right back ankle joint girth in per 0.5 hour 1 time, observe 2.0hr altogether.From as contrast, ankle joint girth (mm) before paw swelling=cause scorching back ankle joint girth-cause is scorching the results are shown in Table 1 with the right back foot of every Mus.
The vigorous tonsillitis of table 1 burdock disappears to the influence of Ovum Gallus domesticus album rat paw edema (X ± SD)
The result shows by table 1, disappear height, middle dosage group of the vigorous tonsillitis of burdock has the obvious suppression effect to 1 one 2h rat Ovum Gallus domesticus album foot swellings, wherein inhibitory action is the strongest behind the 1hr, but effect illustrates that not as positive drug aspirin this product has certain antagonism to the fugitive inflammation that causes of Ovum Gallus domesticus album.
3.1.2, the vigorous tonsillitis of burdock disappears that mice dimethylbenzene auricle edema is influenced
Get 50 mices, be divided into 5 groups at random, 10 of every treated animals, the 1st, 2,3 group is the vigorous tonsillitis of the burdock high, medium and low dosage group that disappears, dosage is respectively 15.6g, 7.8g, 3.9g crude drug/kg (being equivalent to 4 times, 2 times and dose,equivalent respectively), the 4th group of positive medicine aspirin, dosage 0.3g/kg, the 5th group is matched group, each treated animal ig administration every day according to dosage 1 time, continuous 3 times, matched group gives excipient by administration high dose volume.1h after the last administration evenly smears dimethylbenzene 30ul as causing scorching ear in the wide both sides of auris dextra, left ear causes scorching back 2h and cuts two ears in contrast, uses the 9mm card punch, in the facial position punching of same ear, weigh to be equal to the punching auricle, and be the swelling degree with two ear weight differences.The results are shown in Table 2.
The vigorous tonsillitis of the table 2 burdock xylol that disappears causes the influence of mice auricle swelling
Compare with matched group
*P<0.05
*P<0.01
The result shows by table 2, and the vigorous tonsillitis of the burdock 15.6g crude drug/kg dosage that disappears has the effect of obvious suppression auricle edema, and suppression ratio is 19.0%, but effect is not as positive drug aspirin group, in, low dose group also has certain inhibition trend.
3.1.3 the vigorous tonsillitis of burdock disappears to the influence of mouse peritoneal permeability
Get 60 mices, be divided into 5 groups at random, 10 of every treated animals,, the 1st, 2,3 groups is the vigorous tonsillitis of the burdock height that disappears, in, low dose group, dosage is respectively 15.6g, 7.8g, 3.9g crude drug/kg (is equivalent to 4 times of dose,equivalents respectively, 2 times and dose,equivalent), the 4th group of positive medicine aspirin, dosage is 0.30g/kg, the 5th group is matched group, every day, the ig administration was 1 time, the continuous gastric infusion of animal 5 days, in last administration tail vein injection 0.5% azovan blue solution 0.1ml/10g after 1 hour, lumbar injection 0.6% acetic acid normal saline solution 0.2ml/ is only immediately, put to death mice after 20 minutes, cut off the abdominal cavity, use normal saline flushing, collect flushing liquor, transfer to 5ml, centrifugal, get supernatant, survey trap in the 590nm place with 752 type spectrophotometers, survey the OD value, the results are shown in Table 3.
The vigorous tonsillitis of the table 3 burdock Dichlorodiphenyl Acetate that disappears causes the influence of mice capillary permeability
Compare with matched group
*P<0.05
*P<0.01
* *P<0.001
The result shows by table 3, and the vigorous tonsillitis of the burdock high, medium and low three kinds of dosage that disappear all have in various degree inhibitory action to the mouse peritoneal capillary permeability.
3.2 antivirus test
3.2.1 the vigorous tonsillitis of burdock disappears, Radix Astragali injection and guanidine hydrochloride be to In vitro culture people amniotic membrane (FL) cytotoxic assay:
Medicine to be measured is mixed with the solution of 5000 μ g/ml, 2500 μ g/ml, 2000 μ g/ml, 1500 μ g/ml, 1000 μ g/m, 800 μ g/ml, 400 μ g/ml, 200 μ g/ml, 100 μ g/ml, 50 μ g/ml respectively with RPM1640.On people's amniotic membrane (FL) cell, measure maximal non-toxic dosage (TD0) and half toxic dose (TD50) respectively with mtt assay.The TD0 that the vigorous tonsillitis of burdock disappears to people's amniotic membrane (FL) cell is 800 μ g/ml, and Radix Astragali injection is 500mg/ml, and guanidine hydrochloride is 250 μ g/ml (the results are shown in Table 4).
3.2.2. test grouping:
Normal cell matched group, complete virus infected group, positive drug matched group (guanidine hydrochloride), similar medicine matched group (Radix Astragali injection) and test group are established in test.Test group is further divided into 3 groups, begins to establish 800 μ g/ml, 400 μ g/ml, 200 μ g/ml from the vigorous tonsillitis of the burdock TD0 that disappears, and every group is divided into first dosing postoperative infection virus group, infective virus dosing group behind dosing group and the first infective virus simultaneously again.The experimental concentration of similar medicine matched group (Radix Astragali injection) is 500mg/ml.The experimental concentration of positive drug matched group (guanidine hydrochloride) is 250 μ g/ml.
4.2.3 antivirus test: people's amniotic membrane (FL) cell 2.0 * 10
5Cell/0.1ml is inoculated in 96 orifice plate (coxsackie Bs
3Virus, adenovirus type VII), the 0.1ml/ hole; Madin-Darby canine kidney(cell line) (MDCK) (MDCK) 2.0 * 10
5Cell/0.1ml is inoculated in 96 orifice plates.Wait to cultivate 24 hours adherent after, respectively with 100TCID
50Coxsackie B
3(Nancy strain), adenovirus type VII and influenza virus (FM1) infect corresponding host cell, keep inclining in 2 hours, medicine to be measured and positive control medicine are begun to make 3 dilution factors adding cells from maximal non-toxic concentration (TD0), microscopically is observed CPE, destructiveness according to CPE is divided into 4 grades, promptly+---be≤25%++--for≤50%+++-for≤75%++++ is≤100% (table 5,6,7 as a result).
Carry out the virus breeding inhibition test simultaneously, the MTT testing result shows that the vigorous tonsillitis of burdock disappears to coxsackie B
3Virus standard strain Nancy strain, adenovirus type VII strain and influenza virus (FM1) strain all have stronger inhibitory action (seeing Table 4,5,6).More than each test group all carry out t check with the virus control group, prove that each drug test group and virus control group more all have utmost point significant difference level (P<0.001).
3.2.4 conclusion (of pressure testing)
Result of the test shows that the vigorous tonsillitis of burdock disappears to coxsackie B
3Type virus, adenovirus type VII and influenza virus (FM1) all have stronger inhibitory action; Prompting this product may equally also have certain therapeutical effect to the acute pharyngolaryngitis that is caused by coxsackie B type virus, adenovirus and influenza virus.Change of coxsackie b virus, adenovirus and influenza virus are one of main paathogenic factors of acute pharyngolaryngitis.Above-mentioned result of the test shows that obviously the vigorous tonsillitis of burdock disappears and can reduce tonsillitic generation, simultaneously to having certain recovery and protective effect by the damage of the histiocyte due to the viral infection.Because it is that the modern study result has proved its antiviral effect again, so it can reach the effect of the sore-throat relieving that dispels the wind, heat-clearing and toxic substances removing according to necessarily Chinese medical theory and the Chinese medicine II class compound preparation of clinical experience be combined into widely that the vigorous tonsillitis of burdock disappears.
The extracorporeal antivirus effect that the vigorous tonsillitis of burdock disappears act as to be used this Drug therapy acute pharyngolaryngitis clinically certain pharmacodynamics foundation is provided.
3.3 bacteriostatic test
3.3.1, the vigorous tonsillitis of burdock disappears to the inhibitory action [6] of external antibacterial culturing
6 kinds of experimental bacteria are seeded on the common Colombia blood plate, hatch taking-up in 16-18 hour for 37 ℃, with the dilution of physiology salt, making bacterial concentration is 10
6/ ml bacterium, get 10 of sterile test tube during test, medicinal liquid stock solution is diluted by the geometric ratio multiple, after concentration is respectively 0.5g, 0.25g, 0.13g, 0.06g, 0.03g, 0.015g, 0.0075g crude drug/ml, gets 10 every pipes of test tube again and add 1ml medicinal liquid and 0.1ml bacterium liquid, fully behind the mixing, hatched 18 hours for 37 ℃, the result with drug liquid tube whether clarify, to have or not mould collarium be index to liquid level, do not have mould collarium as drug liquid tube clarification, liquid level, then this pipe medicine has antibacterial action; As drug liquid tube is muddy, and liquid level has mould collarium, and then this pipe medicine does not have antibiotic effect.Grow lowest concentration of drug as the minimum inhibitory concentration (MIC) of medicine to this bacterial strain with integral asepsis.The results are shown in Table 8.
The vigorous tonsillitis of table 8 burdock disappears to 6 kinds of experimental bacteria external (test tube method) drug sensitive test
The result shows by table 8, and the vigorous tonsillitis of the burdock different gradient medicinal liquids that disappear all can be measured minimum inhibitory concentration.
3.3.2, to the protective effect of infecting mouse in the staphylococcus aureus body
Staphylococcus aureus (purchase in Nat'l Pharmaceutical ﹠ Biological Products Control Institute bacterium number 26003) is cultivated 37 ℃ hatch taking-up in 16-18 hour, with 10 times of serial dilutions, add the bacteria suspension that 5% concentration gastron (to strengthen virulence) is made different gradients with normal saline simultaneously.
Through trial test, lumbar injection staphylococcus aureus minimum lethal dose (MLD) is 10 under this experimental condition
-2Formal test, get 120 of mices, male and female half and half, body weight 18-20g, be divided into 6 groups at random, every group 20,1st, 2,3,4 groups is the vigorous tonsillitis of burdock four the dosage groups that disappear, and dosage is respectively 31.2g, 15.6g, 7.8g, 3.9g crude drug/kg (8 times, 4 times, 2 times and dose,equivalent being equivalent to dose,equivalent respectively), the 5th group of positive medicine Amoxicillin Capsules group, dosage is 520mg/kg (being equivalent to 2 times of dose,equivalents), the 6th group is matched group, and every day, the ig administration was 2 times, and the upper and lower noon respectively once, successive administration two days is in the 4th administration pneumoretroperitoneum injection 10
-2The staphylococcus aureus suspension, every 0.5ml observes 24h dead animal number, calculates mortality rate and carries out λ
2Statistical procedures the results are shown in Table 9.
Table 9, to the protective effect of (abdominal cavity) infecting mouse in the staphylococcus aureus body
Table 9 is the result show, the vigorous tonsillitis of burdock 31.2g, the 15.6g crude drug/kg dosage that disappears all causes in the mice body staphylococcus aureus and infects the inhibitory action that has in various degree, wherein the high dose group bacteriostasis is the most obvious, learns more by statistics with matched group and handles P<0.01, P<0.05.
3.3.3, the vigorous tonsillitis of burdock disappears to the protective effect of infecting mouse in the pneumonia streptococcus thalline
Streptococcus pneumoniae is cultivated 37 ℃ hatch taking-up in 16-18 hour, with 10 times of serial dilutions, add the bacteria suspension that 5% concentration gastron (to strengthen virulence) is made different gradients simultaneously with normal saline.
Through trial test, lumbar injection streptococcus pneumoniae minimum lethal dose (MLD) is 10 under this experimental condition
-1Formal test, get 90 of mices, male and female half and half, body weight 18-20g is divided into 6 groups at random, 15 every group, 1st, 2,3,4 groups is the vigorous tonsillitis of burdock four the dosage groups that disappear, dosage is respectively 31.2g, 15.6g, 7.8g, 3.9g crude drug/kg, the 5th group of positive medicine Amoxicillin Capsules group, and dosage is 0.52g/kg (being equivalent to 2 times of dose,equivalents), the 6th group is matched group, every day, the ig administration was 2 times, the upper and lower noon each once, successive administration two days, in the 4th administration pneumoretroperitoneum injection streptococcus pneumoniae, every 0.5ml observes 48h dead animal number, calculates mortality rate and carries out λ
2Statistical procedures the results are shown in Table 10.
The protective effect of (abdominal cavity) infecting mouse in the table 10 pair pneumonia streptococcus thalline
The result shows by table 10, and 4 dosage all cause streptococcus pneumoniae and infect the inhibition trend that has in various degree in the mice body, and wherein high, middle dosage group antibacterial action is the most obvious, learns more by statistics with matched group and handles P<0.05.
4.4, separate heat test
4.4.1 disappearing, the vigorous tonsillitis of burdock yeast is caused the influence of rat fever
Get 40 of rats, be divided into 5 groups at random, 8 of every treated animals, 1st, 2,3 groups is the vigorous tonsillitis of the burdock high, medium and low dosage group that disappears, dosage is respectively 10.8g, 5.4g, 2.7g crude drug/kg (being equivalent to 4 times, 2 times and dose,equivalent respectively), the 4th group of positive medicine aspirin, dosage 0.20g/kg, the 5th group is matched group.Each group is according to dosage irritated stomach (ig) administration.Before administration, every rat is surveyed normal body temperature, and record.During test, in every rat skin lower injection 10% young beer yeast suspension 3ml/kg, 6hr after pyrogenicity, dosed administration shown in each treated animal is pressed, the refrigeration function of 0.5hr, 1hr, 1.5hr, 2hr, 3hr the results are shown in Table 11 after observation and the record administration.
Table 11 is the result show, the vigorous tonsillitis of the burdock high dose group that disappears causes that to yeast rat fever has tangible refrigeration function.
3.5 analgesic activity
Dichlorodiphenyl Acetate causes the influence of mouse writhing reaction 3.5.1 the vigorous tonsillitis of burdock disappears
Get 50 mices, be divided into 5 groups at random, 10 of every treated animals, the 1st, 2,3 groups is the vigorous tonsillitis of the burdock height that disappears, in, low dose group, dosage is respectively 15.6g, 7.8g, 3.9g crude drug/kg (is equivalent to 4 times respectively, 2 times and dose,equivalent), the 4th group of positive medicine aspirin, dosage 0.3g/kg, the 5th group is matched group, every day, the ig administration was 1 time, the continuous gastric infusion of animal 5 days (positive drug administration on the same day), 1hr after the last administration, every Mus is by lumbar injection 0.6% acetum 0.1ml/10g (body weight), begin behind the 5min to observe and the interior mouse writhing reaction times of record 10min, the results are shown in Table 12.
The vigorous tonsillitis of the table 12 burdock Dichlorodiphenyl Acetate that disappears causes the influence of mouse writhing reaction
Compare with matched group
*P<0.05
* *P<0.001
Table 12 as can be known, the vigorous tonsillitis of the burdock high, medium and low three kinds of dosage that disappear all can reduce acetic acid and cause the mouse writhing number of times, wherein high dose, middle dosage effect are the strongest.
3.5.2 influence to the reaction of mice hot plate
Get 60 of female mices, 1 is placed on the hot plate at every turn, hot plate temperature is controlled at 55 ± 0.5 ℃, and mice is from being placed on the hot plate to the pain threshold of metapedes required time (s) as this mice occurring licking, allly licks the metapedes time and gives it up less than 5s or greater than 30s or leaper.Will
Garbled 50 mices are divided into 5 groups at random, and 10 every group, according to group dosage, successive administration 7 days is surveyed the pain threshold of 1h, 2h after the last administration, 3h respectively.(60min is still reactionless, and mice is taken out, and calculate with 60s its threshold of pain).
The vigorous tonsillitis of table 13 burdock disappears to the influence of mice hot plate reaction (X ± S)
Compare with matched group
*P<0.05
*P<0.01
Result of the test shows that the vigorous tonsillitis of burdock disappears all has tangible analgesic effect in administration 1h, 2h, 3h to mice.
3.6 to Immune Effects
3.6.1 the influence that the vigorous tonsillitis of burdock disappears carbon granule in the mice body is cleaned up function
Get 50 of mices, male and female half and half are divided into 5 groups at random, every group 10, pressed the continuous gastric infusion of table 14 group dosage 7 days, in 1 hour each Mus of last administration by tail vein injection india ink 0.1ml/10g body weight, then when injection 5min and 15min, get blood 0.025ml from mice vena ophthalmica clump, add among the 1mg/ml sodium carbonate liquor 2ml, shake up, survey the OD value with 752 type spectrophotometer 60nm places, and put to death animal immediately, cut open and get liver, spleen is weighed, calculating K value, α value.The results are shown in Table 14.
The vigorous tonsillitis of table 14 burdock disappears and carbon granule in the mice body is cleaned up the influence (X ± S) of function
Compare with matched group
*P<0.05
*P<0.01
* *P<0.001
As shown in Table 14, the vigorous tonsillitis of burdock 15.6g crude drug/kg, the 7.8g crude drug/kg group that disappears can obviously improve phagocytic function to mononuclear phagocyte, with matched group P<0.001 relatively.
3.7 the vigorous tonsillitis of burdock disappears to the antitussive action of mice
Get 50 of mices, be divided into 5 groups at random, 10 of every treated animals, 1,2,3. group is the vigorous tonsillitis of the burdock high, medium and low dosage group that disappears.Dosage is respectively 15.6g, 7.8g, 3.9g/kg, 4 groups of positive medicine cloperastine groups, and 5 groups is matched group, animal is all irritated stomach (ig) for three days on end, matched group gives with the volume excipient, and after administration in the 3rd day 2hr, with mice according to group in batches (every Mus) put into.Survey in mouthful flask through the carbon dioxide preparation facilities 250ml of improvement, inject bottle with 10ml carbon dioxide quantitative gas, the incubation period (injecting carbon dioxide begins to the required time of mouse cough takes place) of observing cough is causing cough latent period (time s) as measuring observation index, and calculating cough-relieving percentage rate (%).The results are shown in Table 15.
The vigorous tonsillitis of table 15 burdock disappears to the antitussive action of mice (x ± SD)
Compare with matched group
*P<0.05
*P<0.01
The result shows: disappear height, middle dosage of the vigorous tonsillitis of burdock has certain antitussive action to mice, but effect does not have the positive control drug effect obvious.
The result
The result shows by the animal experiment study many index, and the vigorous tonsillitis of burdock disappears to upper respiratory tract easy infection antibacterial staphylococcus aureus 10
-2, streptococcus pneumoniae causes in mice (abdominal cavity) body and infects, disappear height, middle dosage group of the vigorous tonsillitis of burdock has obvious suppression effect (P<0.05), show as and obviously to reduce 24h, 48hr animal dead number, also can prolong the death time of infected mice, but two kinds of dosage inhibitory action are not as the positive reference substance amoxicillin.External drug sensitive test has proved the inhibitory action of this product to 6 kinds of bacterium equally, and from external antibacterial result, this medical instrument has certain facedown bacterial action.
External virus test shows that the vigorous tonsillitis of burdock disappears to coxsackie B
3Type virus, adenovirus type VII and influenza virus (FM1) all have certain inhibitory action; May equally also have certain therapeutical effect to the acute pharyngolaryngitis that causes by coxsackie B type virus, adenovirus and influenza virus.This is for the expulsion pathogenic factor, and releasing table disease is very important.The experimenter thinks that this may be one of pharmacological basis of this medicine.The antiinflammatory experiment shows that this medicine can obviously suppress foot swelling of rat Ovum Gallus domesticus album and Mice Auricle swelling.Also can obviously reduce fresh yeast simultaneously and cause heating rat 0.5h-2.0h body temperature, above-mentioned result of study has confirmed the therapeutical effect of this medicine to primary symptom, and this conforms to clinical efficacy.
By corresponding experimental studies with clinical primary symptom such as antiinflammatory, antiviral, antibacterial, analgesic, pain relieving, raising immunologic functions, every testing index result shows: disappear height, middle dosage group of the vigorous tonsillitis of (1) burdock causes the mouse peritoneal capillary permeability to the foot swelling of rat Ovum Gallus domesticus album, mice dimethylbenzene auricle edema and acetic acid and increases and all have the obvious suppression effect.(2) to coxsackie B
3Type virus, adenovirus type VII and influenza virus (FM1) have certain inhibitory action and inhibition trend (3) respectively and pneumonia infection streptococcus, staphylococcus aureus in the mice body are activated the dead this product of thing significant protective effect is arranged, and extracorporeal bacteria inhibitor test shows that also this product has in various degree drug sensitive reaction to escherichia coli, staphylococcus aureus, klepsiella pneumoniae, streptococcus faecalis, Fu Shi will Hayes etc. simultaneously.(4) can reduce acetic acid and cause the mouse writhing number of times, and increase the mice hot plate pain threshold of reaction, illustrate that this product has certain analgesic effect.(5) beer yeast is caused rat fever the obvious suppression effect is arranged.(6) this medicine also has the phagocytic function of enhancing body mononuclear phagocyte, makes carbon clearance speed accelerate.(7) this product has certain antitussive action.
4, acute toxicity test.
Tonsillitis powder 50% concentration that disappears, give mice ig0.4ml/10g body weight, ig is continuous 2 times in 1 day, and the back was observed 7 days continuously, and animal activity freely, none Mus death, put to death mice in the 8th day, cut open inspection, do not see and organize the obvious abnormal change of internal organs, it is 302.8g crude drug/kg that the result records the maximum dosage of mice ig this product, is equivalent to 704 times of clinical medicine dose.
5, long term toxicity test.
Test is got the Wistar rat and is divided into three dosage groups, a matched group, 80 rats are divided into 4 groups at random, and 20 every group, by setting dosage administration every day 1 time, continuous irrigation stomach 2 months, observe the general state situation of animal behind the medicine, periodic logging body weight, appetite, after administration 2 months and the drug withdrawal 15 days, each puts to death 1/2 animal, and carries out hematology, blood biochemical and reach histopathological examination substantially.The result, during the administration, each treated animal is not seen untoward reaction, weight increase amount, food-intake, hematology and blood biochemical are learned every index all in range of normal value, animal cardinal principle internal organs and main organs histopathologic examination have the inflammatory cell infiltration (relevant with environmental change except that individual animal pulmonary, can recover voluntarily), all the other also no abnormal pathological changes.
Above-mentioned result of study shows, the treatment that is used for acute tonsillitis that disappears of the vigorous tonsillitis of burdock has certain pharmacology foundation, the result of its pharmacological testing conforms to substantially with the result of the clinical prerun of this medicine, and said preparation is observed through acute toxicity test and long term toxicity test, obvious toxic-side effects is seen at the end, and prompting is used this medicine under the dosage of clinical recommendation be safe.Technical study and quality standard research result show that determined preparation technology is reasonable, makes controllable product quality.Three batch samples are by the room temperature reserved sample observing, show that the vigorous tonsillitis of burdock that produces by the preparation technology who determines disappears to keep steady quality constant within a certain period of time.
6, clinical experiment
6.1 diagnostic criteria
6.1.1 tcm diagnosis standard:
(1) main clinical manifestation: pharyngeal pain, increase the weight of when swallowing, companion's fever with aversion to cold or hyperpyrexia, the pharyngeal tonsils redness, there is yellow-white pus point on the surface, does not exceed the pharyngeal tonsils scope, easily wipes.
(2) characteristics of incidence: how relevant with the climate variation, good sending out in child and teenager.
(3) physico-chemical examination: blood leukocytes counting and neutrophil cell number can increase
2, Chinese medical discrimination
Heresy is attacked lung meridian card fever with aversion to cold, and pharyngeal pain is swallowed then aggravation, can be with dry and astringent, and burning sensation, the pharyngeal tonsils redness connects and surrounding tissue, headache, body is tired weak, and malasia brings up phlegm when one coughs, and tongue is red partially, white and thin fur or BOHUANG, floating pulse.
6.1.2 Western medicine diagnose standard
(1) medical history: it is hurried to fall ill, often suffer from cold, tired, tobacco and wine excessively, malnutrition, the low inferior inducement of passive protective physical fitness.
(2) symptom:, general malaise, the shiver with cold heating, fervescence, headache, nape and extremities aching pain, inappetence often has constipation, and pharyngeal pain increases the weight of when swallowing, often with in ear pain.
(3) check: patient's face of acute ill, the buccal flushing, halitosis, thick fur is white, the tonsil redness, the crypts mouth has yellow-white pus point, can merge in flakes, and shape such as pseudomembrane are easily wiped, mandibular lymph node enlargement and tenderness.Lab testing blood leukocytes sum can increase.
6.2 test method:
The vigorous tonsillitis of burdock disappears, and 3 times on the one, each one bag, a week is a course of treatment, carries out efficacy evaluation after finishing the course of treatment.
6.2.1 observation index
Symptom and sign: pharyngalgia, tonsil redness, heating, dry pharynx are scorching hot, headache, situations such as tired weak, the malasia of body, cough, picture of the tongue, pulse condition.
Symptom and sign scalar quantization standard
(1) primary symptom
Normal 0 minute of pharyngalgia
Pharyngalgia was lighter in slight 2 minutes, did not influence and swallowed
4 fens odynophagia of moderate
Severe pain when severe was swallowed in 6 minutes
Red and swollen normal 0 minute of tonsil
Pharyngeal tonsils hyperemia in slight 2 minutes
4 fens pharyngeal tonsils rednesses of moderate have dense point
6 fens pharyngeal tonsils rednesses of severe, the secretions slabbing
(2) inferior disease
Generated heat normal 0 minute
Slight 1 minute<38.5 ℃
38.5~39.5 ℃ of moderates 2 minutes
3 minutes>39.5 ℃ of severes
Scorching hot normal 0 minute of dry pharynx
Pharynx in slight 1 minute is little to be done
2 fens dry pharynxs of moderate are scorching hot
3 fens dry pharynx polydipsia of severe are desired cold drink
Normal 0 minute of General Symptoms
Systemic pain was light or unable in slight 1 minute
2 fens systemic pain of moderate, weak
3 fens systemic pain of severe are weak obvious
Cough does not have 0 fen
There is 1 fen
Tongue is red partially, white and thin fur or BOHUANG, floating pulse: have or not or other.
6.2.2 efficacy assessment standard
Recovery from illness: treat 7 days with interior clinical symptoms and sign disappearance, body temperature, numeration of leukocyte recover normal.The symptom and sign total mark is 0.
Produce effects: treat in 7 days, the curative effect integration reduces 〉=67%, (treatment back total mark reduces 〉=2/3)
Effectively: treat in 7 days, 67% 〉=curative effect integration reduces 〉=33%, (2/3 〉=treatment back total mark reduces 〉=1/3)
Invalid: treat in 7 days, the curative effect integration reduces≤33%, (treatment back total mark reduces≤1/3)
6.2.3 result
6.2.3.1 total effects situation
With reference to the criterion of therapeutical effect of " new Chinese medicine clinical research guideline " acute tonsillitis, the symptom and sign before and after the 30 routine patient is carried out comprehensive evaluation analysis, 14 example recoveries from illness as a result, 10 routine produce effects, 4 is effectively routine, and 2 examples are invalid.Total effective rate is 93.3%.
6.2.3.2 symptom and sign improves situation
The symptom and sign total mark relatively before and after the treatment
Symptom total mark average out to 13.10 ± 3.84 before the 30 routine patient, treatment back symptom total mark is 3.17 ± 4.713, and credit is analysed by statistics, and P<0.01 has significant difference.
Each symptom and sign size scale before and after the treatment