CN105435219A - Preparation method of composite inactivated vaccine for IBD (infection bursal disease) - Google Patents
Preparation method of composite inactivated vaccine for IBD (infection bursal disease) Download PDFInfo
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Abstract
The invention discloses a preparation method of a composite inactivated vaccine for an IBD (infection bursal disease) and belongs to the technical field of animal biological products. The preparation method comprises technological steps as follows: 1), virus reproduction; 2), preparation of the composite inactivated vaccine for the IBD: 1-2 parts of an immunopotentiator is added to each 1,000 ml of an IBDV (infection bursal disease virus) antigen solution checked to be qualified in inactivation, the mixture is uniformly mixed, emulsification packaging is performed on the mixture and a summit livestock water-in-oil-in-water adjuvant in the volume ratio being 3:2, and the composite inactivated vaccine for the IBD is prepared. The novel vaccine adjuvant is adopted, and the prepared vaccine has double characteristics of water in oil and oil in water, combines the sustained release function of an ordinary vaccine and has advantages of joint actions of the water-soluble immunopotentiator, flavone of traditional Chinese medicines, a pain killer and the like. With the adoption of the effective immunopotentiator, the immunity of a body is greatly improved, the cellular immunity capability of the inactivated vaccine is improved, the body is stimulated to generate efficient neutralizing antibodies, and the disease prevention capability of the body is improved.
Description
Technical field
The invention belongs to veterinary biologics technical field, be specifically related to a kind of preparation method of infectious bursal disease compound inactivated vaccine.
Background technology
Infectious bursal disease (Infectionbursaldisease, IBD) be by infectious bursa of Fabricius virus (Infectionbursaldiseasevirus, a kind of high degree in contact sexually transmitted disease of the chicken IBDV) caused and turkey, the young chicken in 3 ~ 12 week age of main infection, this virus is mainly in the lymphocyte internal breeding of fabricius bursa, and damage is in various degree caused to other immune organ, finally causes the fabricius bursa atrophy of chicken and cause the immunosuppressant of chicken.Primary disease can cause losing weight clinically, poor growth and the symptom such as skeletal muscle is hemorrhage, and the chicken being less than 3 week age then can not show obvious Clinical symptoms.But, the chicken bursa of subclinical infection also has some microscopy pathological changes and immunosuppressant phenomenon, has carried out huge economic loss to poultry industrial belt.Primary disease can cause the resistance of chicken to other diseases itself to weaken, and interference is as the immune effect of the vaccine of the diseases such as newcastle, Marek and infectious bronchitis.At present, the kind of IBDV vaccine mainly comprises live vaccine, inactivated vaccine, antigen-antibody complex vaccine, recombinant vaccine etc.According to the infective use of domestic and international live vaccine; although live vaccine can play a part of protective effect; but there is the situation of self fabricius bursa atrophy; thus cause the disorder of self immune system, there is protective rate situation on the low side equally in recombinant vaccine, antigen-antibody complex vaccine in addition.Higher antibody can be produced after inactivated vaccine immunity chicken; effectively can protect body; but fabricius bursa deactivation epidemic disease domestic at present, because its immune effect is poor or production cost is high, is not extensively promoted, the most or expensive import vaccine that each macrospecies chicken house uses.Newcastle, infectious bursal disease are the two kinds of serious infectious diseases causing chickling high mortality, seriously govern growing up healthy and sound of poultry husbandry.Carrying out the epidemic preventing working of these two kinds of diseases, particularly the epidemic preventing working of 1 Japanese instar chickling, is the prerequisite ensureing that poultry husbandry develops in a healthy way.Therefore, develop and a kind ofly can be used for into chicken and can be used for again Immune Profile In Chicks and prevent the safe and effective vaccine of these two kinds of diseases very urgent.For poultry husbandry the problems referred to above, the proposition of this project can solve in poultry husbandry the immunoprophylaxis difficult problem becoming chicken and chicken Newcastle disease, infectious bursal disease, reduces the popular of disease in industry, increases the rate of animals delivered to the slaughter-house, for the poultry husbandry of China escorts.
Water-in-oil-in-water emulsion adjuvant has the feature of aluminium glue adjuvant, oil-in-water adjuvant and Water-In-Oil adjuvant, compared with water-in-oil emulsion adjuvant, its antigen concentration prepared required for vaccine reduces greatly, in addition because it has the characteristic of oil-in-water adjuvant, make it can add water miscible immunostimulant, herbal polysaccharide and analgesics (vitamin), can alleviating pain when vaccine organism, reduce Animal stress, simultaneously immunostimulant, herbal polysaccharide also have raising immune effect, and the function such as prevent disease.
Vitamin is requisite organic compound in organism metabolism.The chemical plant that body is very complicated just as one, constantly carries out various biochemical reaction.Its reaction has substantial connection with the catalytic action of enzyme.Enzyme will produce activity, and coenzyme must be had to participate in.Known many vitamin are the coenzyme of enzyme or the ingredient of coenzyme.Therefore, vitamin is the important substance maintaining and regulate body homergy.
Flavone compound is the polyatomic phenol material that occurring in nature exists, and is also one of main active in nature medicinal plants.It refers to have 15 carbon atoms and with the three ring natural organic matters that the mode of C6-C3-C6 is formed, be the secondary metabolism product that plant produces in long-term natural selection, now isolation identification have kind more than 4000.It is extensively present in fruit and vegerable, Chinese herbal medicine, have no side effect, it has the medicine healthy sofa functions such as significant scavenger interior free yl, aging resistance, mutation, Adjust-blood lipid blood pressure lowering, it is the natural organic oxidation-resistant agent that a class has DEVELOPMENT PROSPECT, these anti-oxidation active substances can reduce and scavenger interior free yl, have prophylactic effect.
Bursopoietin (bursin, BS) is a kind of active kyrine material separated from birds distinctive humoral immunization central lymphoid organ's fabricius bursa (Bursaoffabricius), and its structure is L-Lys-His-Gly-NH2.Research shows, BS can promote differentiation, the propagation of birds and mammalian B lymphocytes precursor, second message,second messenger cAMP and cGMP level in B cell system Daudi cell can be improved, accelerate DNA in B cell and be transcribed into the speed of mRNA, promote the generation of B cell internal protein, thus B cell is produced and the reinforcement of secretory antibody ability.
The present invention is based on above-mentioned technical background, propose a kind of preparation method of infections chicken cloacal bursa compound inactivated vaccine, this vaccine adds the appropriate immunostimulant that contains by infections chicken cloacal bursa antigen and makes, and dosage form is water-in-oil-in water.The compound inactivated vaccine prepared by the method, because it has oil-in-water, Water-In-Oil two kinds of characteristics, the immunostimulants such as water miscible vitamin, bursopoietin, Chinese medicine flavone and analgesics is made to be easy to dissolve, be beneficial to animal body to absorb, can Animal stress be reduced during immunity, reduce animal suffering, can animal welfare be improved; Immunostimulant can improve the cell of live vaccine greatly simultaneously, produces higher neutralizing antibody, significantly improves protective rate.
Summary of the invention
For prior art Problems existing, the object of the invention is to design the technical scheme of the preparation method that a kind of infectious bursal disease compound inactivated vaccine is provided.
The preparation method of described a kind of infectious bursal disease compound inactivated vaccine, is characterized in that comprising following processing step:
1) virus multiplication
(1) the going down to posterity of DF-1 cell
Get and cultivate within 24-48 hour, cellular morphology is good, is evenly distributed and covers with the cell bottle of monolayer, discarding cell culture fluid, add trypsin solution digestion in Tissue Culture Flask; Outwell cell dissociation buffer, in cell bottle, then add the DMEM/F12 cell growth medium containing 10% hyclone, with suction pipe featheriness cell surface, make cell detachment and be uniformly dispersed, then adding cell growth medium and go down to posterity by 1:3-4;
(2) inoculate
Grow up to after monolayer until 24-48 hour cell and carry out changing liquid and connecing poison, get production seed culture of viruses, grown up to the DF-1 cell of good monolayer by 1% inoculation of cell maintenance medium amount, put 37 DEG C and continue to cultivate, cell maintenance medium is the DMEM/F12 solution containing 2% hyclone;
(3) gather in the crops
Meet the rear 24-48 hour of poison, harvesting liquid when cytopathy reaches more than 75%;
2) infectious bursal disease compound inactivated vaccine preparation
The virus liquid obtained in step 1) is carried out steriling test, viral level measures, carry out formalin-inactivated after the assay was approved, deactivation after the assay was approved, 1 ~ 2 part of immunostimulant mix homogeneously is added in the chicken infectivity bursa of Fabricius virus antigen liquid that every 1000ml deactivation is up to the standards, every portion of immunostimulant contains Chinese medicine flavone 10 ~ 20g, vitamin combination 3 ~ 6g and bursopoietin 10 ~ 20mg, then with Sang meter Te domestic animal W/O/W adjuvant by volume 3:2 carry out emulsifying subpackage and make infectious bursal disease compound inactivated vaccine, above-mentioned Chinese medicine flavone is by Flos Chrysanthemi 10 ~ 30 parts, 10 ~ 30 parts, the Radix Rehmanniae, Fructus Viticis 20 ~ 40 parts, Radix Gentianae 10 ~ 20 parts, Radix Scutellariae 10 ~ 30 parts, extract in the Chinese medicine composition that Rhizoma Smilacis Glabrae 20 ~ 30 parts and Radix Scrophulariae 10 ~ 30 parts are formed and obtain.
The preparation method of described a kind of infectious bursal disease compound inactivated vaccine, is characterized in that described Chinese medicine flavone is obtained by following steps:
A, press Flos Chrysanthemi 10 ~ 30 parts, 10 ~ 30 parts, the Radix Rehmanniae, Fructus Viticis 20 ~ 40 parts, Radix Gentianae 10 ~ 20 parts, Radix Scutellariae 10 ~ 30 parts, Rhizoma Smilacis Glabrae 20 ~ 30 parts, Radix Scrophulariae 10 ~ 30 parts takes each raw material of Chinese medicine, chopping, clean after spend the night by cold water soak, add the purified water of raw material weight 15 times again, water-bath to 90 DEG C, and remain on 90 DEG C, boil 2 hours, boil in process every 10 minutes and stir once;
B, discard a Chinese medicine slag, collect medicinal liquid, after room temperature cooling through 10000rpm centrifugal 10 minutes, discard precipitation, collect supernatant;
C, supernatant in b is carried out precipitate with ethanol, namely supernatant obtains rough Chinese medicine flavone;
D, Chinese medicine flavone rough in c is carried out reduced vacuum concentrate, concentrate time sterilized water for injection dilution cleaning, again carries out reduced vacuum and concentrates, after three times like this, degerming finally by 0.22um membrane filtration, then namely obtains Chinese medicine flavone through lyophilisation.
The preparation method of described a kind of infectious bursal disease compound inactivated vaccine, it is characterized in that described bursopoietin is obtained by following steps: fabricius bursa tissue is rejected fascia and fatty tissue, the cold PBS of pH7.2 sterilizing cleans, the cold PBS of pH7.2 sterilizing is added in 1:1 ratio, high-speed homogenization is carried out in tissue refiner, the trypsin accounting for 2.5% weight is added in homogenate, multigelation 3 times, the centrifugal 20min of 12000rpm, abandon precipitation, the ultrafilter membrane of supernatant 1000da molecular cut off carries out ultrafiltration, under film, liquid is degerming through 0.22um membrane filtration, filtered solution is bursopoietin crude extract, crude extract obtains bursopoietin through lyophilisation.
The preparation method of described a kind of infectious bursal disease compound inactivated vaccine, is characterized in that in described vitamin combination containing vitamin D2 0 ~ 40%, vitamin E 30 ~ 60% and vitamin B2 10 ~ 30%.
The preparation method of described a kind of infectious bursal disease compound inactivated vaccine, is characterized in that in described vitamin combination containing vitamin D2 5 ~ 35%, vitamin E 35 ~ 55% and vitamin B2 15 ~ 30%.
The preparation method of described a kind of infectious bursal disease compound inactivated vaccine, is characterized in that described Chinese medicine flavone obtains by extracting in Flos Chrysanthemi 15 ~ 25 parts, 15 ~ 25 parts, the Radix Rehmanniae, Fructus Viticis 25 ~ 35 parts, Radix Gentianae 12 ~ 18 parts, Radix Scutellariae 15 ~ 25 parts, Rhizoma Smilacis Glabrae 22 ~ 26 parts and Radix Scrophulariae 15 ~ 25 parts of Chinese medicine compositions formed.
In the present invention, Sang meter Te domestic animal W/O/W adjuvant is by Zhuhai Guo Nian bio tech ltd production and sales.
The present invention has following beneficial effect:
1. present invention employs new vaccine adjuvant, prepared vaccine has Water-In-Oil, oil-in-water double grading, has had the coefficient advantage such as slow releasing function and water solublity immunostimulant, Chinese medicine flavone, analgesics of vaccine concurrently.
2. present invention employs effective immunostimulant, increased substantially the immunity of body, improve inactivated vaccine cell immunocompetent, stimulate body to produce efficient neutralizing antibody, improve the prophylactic ability of body.
Detailed description of the invention
In order to make the present invention easier to understand, below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention and place restrictions on scope of the present invention, NM specific experiment method in the following example, experimental technique carries out routinely usually.
Prepared by embodiment 1 chicken infectivity bursa of Fabricius virus suspension
(1) the going down to posterity of DF-1 cell
Get and cultivate within 24-48 hour, cellular morphology is good, is evenly distributed and covers with the cell bottle of monolayer, discarding cell culture fluid, add the digestion of appropriate trypsin solution in Tissue Culture Flask; Outwell cell dissociation buffer, in cell bottle, then add the DMEM/F12 cell growth medium containing 10% hyclone, with suction pipe featheriness cell surface, make cell detachment and be uniformly dispersed, then adding cell growth medium and go down to posterity by 1:3-4.
(2) inoculate
Carry out after 24-48 hour cell grows up to monolayer changing liquid and connecing poison, get production seed culture of viruses (infectious bursa of Fabricius virus HQ strain), grown up to the DF-1 cell of good monolayer by 1% inoculation maintaining liquid measure, put 37 DEG C and continue to cultivate.Cell maintenance medium is the DMEM/F12 solution containing 2% hyclone.
(3) gather in the crops
Meet the rear 24-48 hour of poison, harvesting liquid when cytopathy reaches more than 75%.
(4) detection of virus liquid
Viral level measures: measure viral suspension in (3) by DF-1 cell line, every 0.1ml should be not less than 10
7.5tCID
50.
Steriling test: undertaken by " Chinese veterinary pharmacopoeia ", should without bacterial growth.
The preparation of embodiment 2 Chinese medicine flavone
A, press Flos Chrysanthemi 25 parts, 25 parts, the Radix Rehmanniae, Fructus Viticis 35 parts, Radix Gentianae 18 parts, Radix Scutellariae 25 parts, Rhizoma Smilacis Glabrae 26 parts, Radix Scrophulariae 25 parts takes each raw material of Chinese medicine, chopping, clean after spend the night by cold water soak, add the purified water of raw material weight 15 times again, water-bath to 90 DEG C, and remain on 90 DEG C, boil 2 hours, boil in process every 10 minutes and stir once;
B, discard a Chinese medicine slag, collect medicinal liquid, after room temperature cooling through 10000rpm centrifugal 10 minutes, discard precipitation, collect supernatant;
C, supernatant in b is carried out precipitate with ethanol, namely supernatant obtains rough Chinese medicine flavone;
D, supernatant in c is carried out reduced vacuum concentrate, concentrate time sterilized water for injection dilution cleaning, again carries out reduced vacuum and concentrates, after three times like this, degerming finally by 0.22um membrane filtration, then namely obtains Chinese medicine flavone through lyophilisation.
Can press Flos Chrysanthemi 10 parts in this embodiment 2,10 parts, the Radix Rehmanniae, Fructus Viticis 20 parts, Radix Gentianae 10 parts, Radix Scutellariae 10 parts, Rhizoma Smilacis Glabrae 20 parts, Radix Scrophulariae 10 parts takes each raw material of Chinese medicine; Also can press Flos Chrysanthemi 30 parts, 30 parts, the Radix Rehmanniae, Fructus Viticis 40 parts, Radix Gentianae 20 parts, Radix Scutellariae 30 parts, Rhizoma Smilacis Glabrae 30 parts, Radix Scrophulariae 30 parts takes each raw material of Chinese medicine
The preparation of embodiment 3 bursopoietin
A, get Jian Kangfashi lens capsule tissue, fabricius bursa tissue is rejected fascia and fatty tissue, the cold PBS of pH7.2 sterilizing cleans.
B, weighing fabricius bursa weight, in 1:1(W:V) ratio adds the cold PBS of pH7.2 sterilizing, in tissue refiner, carry out high-speed homogenization.
Add the trypsin of 2.5% in c, homogenate, multigelation 3 times, the centrifugal 20min of 12000rpm, abandons precipitation.
The ultrafilter membrane of d, supernatant 1000da molecular cut off carries out ultrafiltration, and under film, liquid is bursopoietin crude extract.
E, by crude extract in d by 0.22um membrane filtration degerming, then namely obtain bursopoietin through lyophilisation.
Prepared by embodiment 4 immunostimulant
Take Chinese medicine flavone 15g, vitamin combination 3g(wherein vitamin D 0.9g, vitamin e1 .2g, vitamin B2 0.9g), bursopoietin 15mg; Above-mentioned each component mix homogeneously is used as portion metering.Wherein herbal polysaccharide is obtained by the step according to embodiment 2, and Chinese medicine flavone obtains according to the step of embodiment 2, and bursopoietin obtains according to the step of embodiment 3.
Prepared by embodiment 5 immunostimulant
Take Chinese medicine flavone 20g, vitamin combination 6g(wherein vitamin D1 .2g, vitamin E 3.6g, vitamin B2 1.2g), bursopoietin 20mg; Above-mentioned each component mix homogeneously is used as portion metering.Wherein Chinese medicine flavone obtains according to the step of embodiment 2, and bursopoietin obtains according to the step of embodiment 3.
Prepared by embodiment 6 immunostimulant
Take Chinese medicine flavone 10g, vitamin combination 5g(wherein vitamin D1 .8g, vitamin E2 .5g, vitamin B2 0.7g), bursopoietin 12mg; Above-mentioned each component mix homogeneously is used as portion metering.Wherein Chinese medicine flavone obtains according to the step of embodiment 2, and bursopoietin obtains according to the step of embodiment 3.
The preparation method of embodiment 7 one kinds of infectious bursal disease compound inactivated vaccines
Under a, aseptic condition, antigen liquid after chicken infectivity bursa of Fabricius virus deactivation is added the immunostimulant 1 or 2 parts prepared in embodiment 4,5 or 6 by every 1000ml, mix homogeneously, makes immunostimulant dissolve completely.
B, 3 deals are added in emulsator containing the chicken infectivity bursa of Fabricius virus inactivation antigen liquid of immunostimulant, regulate emulsator rotor speed to be 1600rpm, the Sang meter Te water profit adjuvant of 2 deals is slowly added the antigen liquid in stirring.
C, when oil phase adds, regulating rotary rotor speed is 3200rpm, continues to stir 3-5 minute.
After d, emulsifying completely, make infectious bursal disease compound inactivated vaccine by aseptic subpackaged for vaccine after emulsifying.
F, product inspection
(1) assay
Carry out related check according to " People's Republic of China's veterinary drug allusion quotation " (2010 editions) related request to finished product, assay is in table 1:
Table 1 product inspection result
(2) immune effect of vaccine compares
Get 100 2 monthly age SPF chickens, be divided into two groups, experimental group and matched group.Experimental group immunity compound inactivated vaccine of the present invention, the common inactivated vaccine of composite immune reinforcing agent is not added in matched group immunity, and immunity plays blood sampling its antibody titer of detection for latter 3 days and agar diffusion is tired.
Claims (6)
1. a preparation method for infectious bursal disease compound inactivated vaccine, is characterized in that comprising following processing step:
1) virus multiplication
(1) the going down to posterity of DF-1 cell
Get and cultivate within 24-48 hour, cellular morphology is good, is evenly distributed and covers with the cell bottle of monolayer, discarding cell culture fluid, add trypsin solution digestion in Tissue Culture Flask; Outwell cell dissociation buffer, in cell bottle, then add the DMEM/F12 cell growth medium containing 10% hyclone, with suction pipe featheriness cell surface, make cell detachment and be uniformly dispersed, then adding cell growth medium and go down to posterity by 1:3-4;
(2) inoculate
Grow up to after monolayer until 24-48 hour cell and carry out changing liquid and connecing poison, get production seed culture of viruses, grown up to the DF-1 cell of good monolayer by 1% inoculation of cell maintenance medium amount, put 37 DEG C and continue to cultivate, cell maintenance medium is the DMEM/F12 solution containing 2% hyclone;
(3) gather in the crops
Meet the rear 24-48 hour of poison, harvesting liquid when cytopathy reaches more than 75%;
2) infectious bursal disease compound inactivated vaccine preparation
The virus liquid obtained in step 1) is carried out steriling test, viral level measures, carry out formalin-inactivated after the assay was approved, deactivation after the assay was approved, 1 ~ 2 part of immunostimulant mix homogeneously is added in the chicken infectivity bursa of Fabricius virus antigen liquid that every 1000ml deactivation is up to the standards, every portion of immunostimulant contains Chinese medicine flavone 10 ~ 20g, vitamin combination 3 ~ 6g and bursopoietin 10 ~ 20mg, then with Sang meter Te domestic animal W/O/W adjuvant by volume 3:2 carry out emulsifying subpackage and make infectious bursal disease compound inactivated vaccine, above-mentioned Chinese medicine flavone is by Flos Chrysanthemi 10 ~ 30 parts, 10 ~ 30 parts, the Radix Rehmanniae, Fructus Viticis 20 ~ 40 parts, Radix Gentianae 10 ~ 20 parts, Radix Scutellariae 10 ~ 30 parts, extract in the Chinese medicine composition that Rhizoma Smilacis Glabrae 20 ~ 30 parts and Radix Scrophulariae 10 ~ 30 parts are formed and obtain.
2. the preparation method of a kind of infectious bursal disease compound inactivated vaccine as claimed in claim 1, is characterized in that described Chinese medicine flavone is obtained by following steps:
A, press Flos Chrysanthemi 10 ~ 30 parts, 10 ~ 30 parts, the Radix Rehmanniae, Fructus Viticis 20 ~ 40 parts, Radix Gentianae 10 ~ 20 parts, Radix Scutellariae 10 ~ 30 parts, Rhizoma Smilacis Glabrae 20 ~ 30 parts, Radix Scrophulariae 10 ~ 30 parts takes each raw material of Chinese medicine, chopping, clean after spend the night by cold water soak, add the purified water of raw material weight 15 times again, water-bath to 90 DEG C, and remain on 90 DEG C, boil 2 hours, boil in process every 10 minutes and stir once;
B, discard a Chinese medicine slag, collect medicinal liquid, after room temperature cooling through 10000rpm centrifugal 10 minutes, discard precipitation, collect supernatant;
C, supernatant in b is carried out precipitate with ethanol, namely supernatant obtains rough Chinese medicine flavone;
D, Chinese medicine flavone rough in c is carried out reduced vacuum concentrate, concentrate time sterilized water for injection dilution cleaning, again carries out reduced vacuum and concentrates, after three times like this, degerming finally by 0.22um membrane filtration, then namely obtains Chinese medicine flavone through lyophilisation.
3. the preparation method of a kind of infectious bursal disease compound inactivated vaccine as claimed in claim 1, it is characterized in that described bursopoietin is obtained by following steps: fabricius bursa tissue is rejected fascia and fatty tissue, the cold PBS of pH7.2 sterilizing cleans, the cold PBS of pH7.2 sterilizing is added in 1:1 ratio, high-speed homogenization is carried out in tissue refiner, the trypsin accounting for 2.5% weight is added in homogenate, multigelation 3 times, the centrifugal 20min of 12000rpm, abandon precipitation, the ultrafilter membrane of supernatant 1000da molecular cut off carries out ultrafiltration, under film, liquid is degerming through 0.22um membrane filtration, filtered solution is bursopoietin crude extract, crude extract obtains bursopoietin through lyophilisation.
4. the preparation method of a kind of infectious bursal disease compound inactivated vaccine as claimed in claim 1, is characterized in that in described vitamin combination containing vitamin D2 0 ~ 40%, vitamin E 30 ~ 60% and vitamin B2 10 ~ 30%.
5. the preparation method of a kind of infectious bursal disease compound inactivated vaccine as claimed in claim 1, is characterized in that in described vitamin combination containing vitamin D2 5 ~ 35%, vitamin E 35 ~ 55% and vitamin B2 15 ~ 30%.
6. the preparation method of a kind of infectious bursal disease compound inactivated vaccine as claimed in claim 1, is characterized in that described Chinese medicine flavone obtains by extracting in Flos Chrysanthemi 15 ~ 25 parts, 15 ~ 25 parts, the Radix Rehmanniae, Fructus Viticis 25 ~ 35 parts, Radix Gentianae 12 ~ 18 parts, Radix Scutellariae 15 ~ 25 parts, Rhizoma Smilacis Glabrae 22 ~ 26 parts and Radix Scrophulariae 15 ~ 25 parts of Chinese medicine compositions formed.
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