CN1029857C - Preparation method of bacteroides microecological preparation - Google Patents
Preparation method of bacteroides microecological preparation Download PDFInfo
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- CN1029857C CN1029857C CN 90102847 CN90102847A CN1029857C CN 1029857 C CN1029857 C CN 1029857C CN 90102847 CN90102847 CN 90102847 CN 90102847 A CN90102847 A CN 90102847A CN 1029857 C CN1029857 C CN 1029857C
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- bacteroides
- bacterial strain
- bacterioide
- fragilis
- active bacteria
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- 238000002360 preparation method Methods 0.000 title abstract description 5
- 241000606125 Bacteroides Species 0.000 title description 2
- 230000001580 bacterial effect Effects 0.000 claims abstract description 10
- 238000000855 fermentation Methods 0.000 claims abstract description 6
- 230000004151 fermentation Effects 0.000 claims abstract description 6
- 241000894006 Bacteria Species 0.000 claims description 21
- 239000000203 mixture Substances 0.000 claims description 18
- 238000009472 formulation Methods 0.000 claims description 13
- 241000606124 Bacteroides fragilis Species 0.000 claims description 7
- 244000005700 microbiome Species 0.000 claims description 6
- 238000011081 inoculation Methods 0.000 claims description 4
- 210000004185 liver Anatomy 0.000 claims description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims description 2
- 239000001888 Peptone Substances 0.000 claims description 2
- 108010080698 Peptones Proteins 0.000 claims description 2
- 235000015278 beef Nutrition 0.000 claims description 2
- 235000021336 beef liver Nutrition 0.000 claims description 2
- 229940041514 candida albicans extract Drugs 0.000 claims description 2
- 239000008103 glucose Substances 0.000 claims description 2
- 238000007654 immersion Methods 0.000 claims description 2
- 238000001802 infusion Methods 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 235000019319 peptone Nutrition 0.000 claims description 2
- 235000015277 pork Nutrition 0.000 claims description 2
- 229910052708 sodium Inorganic materials 0.000 claims description 2
- 239000011734 sodium Substances 0.000 claims description 2
- GNBVPFITFYNRCN-UHFFFAOYSA-M sodium thioglycolate Chemical compound [Na+].[O-]C(=O)CS GNBVPFITFYNRCN-UHFFFAOYSA-M 0.000 claims description 2
- 239000012138 yeast extract Substances 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 7
- 241001465754 Metazoa Species 0.000 abstract description 6
- 230000000968 intestinal effect Effects 0.000 abstract description 5
- 208000019637 Infantile Diarrhea Diseases 0.000 abstract description 4
- 206010057190 Respiratory tract infections Diseases 0.000 abstract description 4
- 206010046306 Upper respiratory tract infection Diseases 0.000 abstract description 4
- 208000012876 acute enteritis Diseases 0.000 abstract description 3
- 235000013361 beverage Nutrition 0.000 abstract description 3
- 238000005516 engineering process Methods 0.000 abstract description 3
- 208000035240 Disease Resistance Diseases 0.000 abstract description 2
- 206010029333 Neurosis Diseases 0.000 abstract description 2
- 230000001154 acute effect Effects 0.000 abstract description 2
- 208000021735 chronic enteritis Diseases 0.000 abstract description 2
- 238000011161 development Methods 0.000 abstract description 2
- 244000144972 livestock Species 0.000 abstract description 2
- 208000015238 neurotic disease Diseases 0.000 abstract description 2
- 244000144977 poultry Species 0.000 abstract description 2
- 208000015817 Infant Nutrition disease Diseases 0.000 abstract 1
- 239000003674 animal food additive Substances 0.000 abstract 1
- 239000002537 cosmetic Substances 0.000 abstract 1
- 235000013402 health food Nutrition 0.000 abstract 1
- 230000001071 malnutrition Effects 0.000 abstract 1
- 235000000824 malnutrition Nutrition 0.000 abstract 1
- 208000020029 respiratory tract infectious disease Diseases 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 9
- 235000013601 eggs Nutrition 0.000 description 7
- 201000006549 dyspepsia Diseases 0.000 description 5
- 239000000654 additive Substances 0.000 description 4
- 235000012054 meals Nutrition 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 208000014540 Functional gastrointestinal disease Diseases 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 201000007637 bowel dysfunction Diseases 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 208000008710 Amebic Dysentery Diseases 0.000 description 2
- 206010001986 Amoebic dysentery Diseases 0.000 description 2
- 208000004429 Bacillary Dysentery Diseases 0.000 description 2
- 206010010774 Constipation Diseases 0.000 description 2
- 241000287828 Gallus gallus Species 0.000 description 2
- 206010017915 Gastroenteritis shigella Diseases 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 208000028017 Psychotic disease Diseases 0.000 description 2
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- 230000004060 metabolic process Effects 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 201000005113 shigellosis Diseases 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 208000031648 Body Weight Changes Diseases 0.000 description 1
- 208000004232 Enteritis Diseases 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000010165 autogamy Effects 0.000 description 1
- 230000036760 body temperature Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 230000004579 body weight change Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 244000005709 gut microbiome Species 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 208000023504 respiratory system disease Diseases 0.000 description 1
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- 238000010186 staining Methods 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
A multifunctional bacterial strain BF839 separated from intestinal flora of well-developed infants or young animals is prepared into a live bacterial preparation by a fermentation technology, can increase the growth and development of children, and has good curative effects on preventing and treating acute and chronic enteritis, flora imbalance, upper respiratory infection, neurosis and the like, particularly obvious curative effects on infantile diarrhea and malnutrition. The feed additive can increase the weight and egg yield of livestock and poultry and animals, and enhance disease resistance, and can be used for preparing cosmetics, health food, beverage, etc.
Description
The invention belongs to technical field of bioengineering, particularly relate to a kind of bacteroides fragilis bacterial strain and application thereof that utilizes microbial fermentation technology to cultivate.
In recent years, American-European advanced country is engaged in the scholar of pathogeny microorganism, many people turn to the research to useful normal microorganism, adopt biotechnology to solve the problem that aspects such as agricultural, livestock industry, body-care, disease preventing and treating (comprise and capture the obstinate fort of cancer) and prolongs life exist.Through international online retrieval, the antitumor action document (F55092322,1978) that relates to bifidus bacillus is arranged, but do not see the report of this class strains separation of relevant bacteroides fragilis, cultivation and application.
The microecosystem of human body and immunity, metabolism, nutrition are relevant, decline along with this system of increase at age is residual gradually and wear out.So the people more than 60 years old is weak and easy cancer stricken easily, then deny less than 60 years old healthy and strong people.Purpose of the present invention is intended to isolate the probiotics that some plays an important role the microecosystem normal physiological bacterium in the low age human or animal body, be transplanted in the human or animal body at advanced age, make its recovery and set up the higher new microecosystem of biomass, improve definite value durability factor level, reach and recover or part is recovered the states such as nutrition, metabolism and immunity that low age is had period.
The present invention implements by following step: screen, isolate multi-functional bacteroides fragilis bacterial strain from well-developed baby or low-aged animal intestinal microflora, utilize microbial fermentation technology to make active bacteria formulation, be used to cure multiple disease, preparation protective foods, beverage, fodder additives and makeup etc.
Bacteroides fragilis is one of normal physiological anaerobic bacterium of human body.This bacterial strain has sent China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, preserving number CGMCCN0.0157 May 3 nineteen ninety.Identify that through Institute of Microorganism, Academia Sinica and Epidemiology Organic Inst., China Preventive Medical Academy all tentative is bacteroides fragilis (Bacteroides, the Fragilis) bacterial strain in, called after BF839.Because of limit by domestic equipment and condition, still can not make satisfactory conclusion to ultrastructure and some characteristic thereof of BF839 bacterial strain.China Committee for Culture Collection of Microorganisms's medical science bacterium center suggestion " after calibrating, considers to enroll the national standard Cordycepps ".
Through nearly 10 years studies show that, this bacterium has following function: (1) improves body immunity, increases the immune t-cell activity; (2) help the absorption of human body digest food; (3) some toxicant in the degraded intestines; (4) get rid of alien bacteria and settle down, adjust intestinal flora and distribute at enteron aisle; (5) synthetic multivitamin and protein in enteric cavity; (6) anticancer growth etc.
By hundreds of routine clinical verifications, the BF839 preparation can increase upgrowth and development of children, and control acute chronic enteritis, flora imbalance, upper respiratory tract infection and neurosis etc. are all had better curative effect, and is particularly remarkable to infantile diarrhea, malnutritive curative effect especially.Can increase poultry, the weight of animals, egg productivity and strengthen disease resistance as fodder additives, also can be used for preparing makeup, protective foods and beverage etc.
BF839 gets from well-developed baby intestinal flora, puts in the anaerobically fermenting substratum of autogamy fermentation and makes active bacteria formulation.Anaerobically fermenting substratum composition is as follows:
Peptone 0.5-2.5%, glucose 0.2-0.5%, halfcystine 0.001-0.5%, NaCl10.2-0.5%, Na
2HPO
40.1-0.3%, sodium thioglycollate 0.02-0.1%, beef liver or pork liver immersion liquid 30-60%, beef infusion broth 30-60%, yeast extract paste 0.1-0.8%(weight percent).Said BF839 strain culturing temperature is to approach the scope of Mammals body temperature, and optimum temps is 37 ℃, constant temperature 32-40 ℃ through 12-72 hour fermentation culture, 3500 rev/mins centrifugal removes supernatant liquor, survey bacterium uses.
The embodiment of the invention is as follows:
Embodiment one: the BF839 inoculation in 10 liter anaerobically fermenting substratum, was cultivated 24 hours for 37 ℃, and recording every milliliter of viable count is 2084 * 10
6=10
8.2(press log10
n/ ml calculates) 3000-4000 rev/min centrifugal supernatant liquor, washing, the diluted for use of going.
Embodiment two: the BF839 inoculation in 10 liter anaerobically fermenting substratum, was cultivated 24 hours for 37 ℃, and recording every milliliter of viable count is 588 * 10
8=10
9.8(press log10
n/ ml calculates) 3000-4000 rev/min centrifugal supernatant liquor, washing, the diluted for use of going.
Embodiment three: the BF839 inoculation in 10 liter anaerobically fermenting substratum, was cultivated 30 hours for 37 ℃, and recording every milliliter of viable count is 1132 * 10
7=10
9.1(press log10
n/ ml calculates).3000-4000 rev/min of centrifugal supernatant liquor, washing, the diluted for use of going.
Embodiment four: BF839 active bacteria formulation 10ml(viable count 4,000,000,000/ml) of early 10 minutes after the meal clothes next day of boarding at the nursery children for the 3-6 of kindergarten year, observed altogether 11.5 months, the result shows and takes height and the body weight (table 1 that the BF839 active bacteria formulation has significantly increased children, table 2), (table 1, table 2 are seen the literary composition back)
Embodiment five: the BF839 active bacteria formulation is used to improve immunization programs for children power, give the BF839 active bacteria formulation 10ml(viable count 4,000,000,000/ml) of children's day early 10 minutes after the meal clothes of boarding at the nursery in the 3-6 of kindergarten year, employing esterase staining mensuration after 11 months, result show takes the immunologic function (table 3) that the BF839 active bacteria formulation has significantly increased children.(table 3 is seen the literary composition back)
Embodiment six: the BF839 active bacteria formulation is used for the treatment of enteritis, bowel dysfunction, indigestion.236 patients such as acute enteritis, cureless chronic intestinal inflammations (part is made a definite diagnosis through enteroscopy), bowel dysfunction, general liver property and the senile maldigestion that the standard routinely of giving is made a definite diagnosis, infantile diarrhea, amebic dysentery, depressive psychosis, bacillary dysentery, constipation early service once the BF839 active bacteria formulation every day after meal, each 20ml(viable count 4,000,000,000/ml), the efficient 98.3%(table 5 that reaches).(table 4 is seen the literary composition back)
Embodiment seven: the BF839 active bacteria formulation is used to prevent and treat upper respiratory tract infection.BF839 active bacteria formulation 10ml(viable count 2,000,000,000/ml) of early 10 minutes after the meal clothes was observed 11.5 months altogether next day of boarding at the nursery children for the 3-6 of kindergarten year, and the result shows the effect (table 5) that prevention child upper respiratory disease is arranged.(table 5 is seen the literary composition back)
Embodiment eight: with 120 of the poulty house laying hens with age in days of the same race, under identical living condition, use forage feed of the same race, stochastic sampling is divided into each 60 of test group and control groups.Test group is taken bacterium several 2,000,000,000 every day by every chicken amount is admixed in the feed feeding with BF839 viable bacteria additive and is suffered all.31 days trial periods, continued to observe 21 days behind hello the additive with stopping.In amounting to 52 days, 60 egg productivitys of test group increase 17.53Kg(table 6)
Body weight change result behind the table 1 children taking BF839
Continue Time of Administration (moon) n1(test group) the n2(control group) T value P value
2 64 54 2.5 ＜0.05
5 53 46 2.97 ＜0.01
7.5 59 56 3.58 ＜0.01
Height result of variations behind the table 2 children taking BF839
Continue Time of Administration (moon) n1(test group) the n2(control group) T value P value
6 53 46 1.78 ＞0.05
11.5 44 46 2.90 ＜0.01
Table 3 T raji cell assay Raji result
The n1(test group) T value P value n2(control group)
Group experiment 45 38 4.18＜0.01
Pair test 30 30 3.25＜0.01
Table 4 is taken the curative effect of BF839 to intestinal tract disease
The example number each one take bottle number rehabilitation take a turn for the better invalid
Chronic intestinal inflammations 81 1-10 74 61
Acute enteritis 17 1-3 17
Bowel dysfunction 22 1-6 16 42
General maldigestion 32 1-4 30 2
Liver property maldigestion 8 4-10 53
Senile maldigestion 6 3-10 6
Infantile diarrhea 58 1-3 58
Amebic dysentery 141
Depressive psychosis 343
Bacillary dysentery 444
Constipation 444
Sum 236 214 18 4
Percentage 100% 90.7% 7.6% 1.7%
Table 5 BF839 is to the prophylactic effect of upper respiratory tract infection of children
Continue Time of Administration (moon) n1(test group) the n2(control group) T value P value
11.5 63 55 3.12 ＜0.01
Table 6 BF839 increases the egg productivity of chicken
The average egg number of the sum (only) of laying eggs (only/day) total amount of laying eggs (Kg) average egg yield (Kg/ days)
Test group 2,379 45.75 143.09 2.7
Control group 2,124 40.85 125.56 2.42
Claims (2)
1, a kind of bacterioide active bacteria formulation, this bacterioide (depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center; Preserving number CGMCCN0.0157; Specific name bacteroides fragilis Bacteroides, Fragilis called after BF839), be to have a strong fermentation character and a FFI bacterial strain in the bacillus fragilis, it is characterized in that the BF839 inoculation in substratum; Peptone 0.5-2.5%, glucose 0.2-0.5%, halfcystine 0.001-0.5%, NaCl0.2-0.5%, Na
2HPO
40.1-0.3%, sodium thioglycollate 0.02-0.1%, beef liver or pork liver immersion liquid 30-60%, beef infusion broth 30-60%, yeast extract paste 0.1-0.8% (weight percent) ferments through 12-72 hour at constant temperature 32-40 ℃, filters, washs, dilutes, surveys bacterium then and use.
2, bacterioide active bacteria formulation according to claim 1 is characterized in that BF839 bacterial strain constant temperature culture temperature is 37 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 90102847 CN1029857C (en) | 1990-05-03 | 1990-05-03 | Preparation method of bacteroides microecological preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 90102847 CN1029857C (en) | 1990-05-03 | 1990-05-03 | Preparation method of bacteroides microecological preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1056314A CN1056314A (en) | 1991-11-20 |
| CN1029857C true CN1029857C (en) | 1995-09-27 |
Family
ID=4877730
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 90102847 Expired - Fee Related CN1029857C (en) | 1990-05-03 | 1990-05-03 | Preparation method of bacteroides microecological preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1029857C (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103082294A (en) * | 2013-01-21 | 2013-05-08 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparation of composition for treating diarrhea |
| CN103146620A (en) * | 2013-03-25 | 2013-06-12 | 广州知光生物科技有限公司 | Bacteroides fragilis with characteristics of probiotics |
| WO2019095508A1 (en) * | 2017-11-17 | 2019-05-23 | 中山大学 | Composition for enhancing immune function of t cells and preparation method therefor |
| CN113151083A (en) * | 2021-04-21 | 2021-07-23 | 中国疾病预防控制中心传染病预防控制所 | Bacteroides dorsalis probiotics and application thereof in preparation of influenza treatment or prevention medicine |
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| CN103120705A (en) * | 2013-01-21 | 2013-05-29 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparation of composition for treating constipation |
| CN103142657A (en) * | 2013-03-18 | 2013-06-12 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparing composition for preventing and treating hypercholesteremia |
| CN103142658A (en) * | 2013-03-18 | 2013-06-12 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparing composition for preventing and treating acute radiation enteritis |
| CN103156887B (en) * | 2013-03-18 | 2015-06-24 | 广州知易生物科技有限公司 | Application of bacteroides fragilis in preparation of composition for promoting bifidobacterium and lactobacillus to grow |
| CN103156888A (en) * | 2013-03-18 | 2013-06-19 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparation of composition for treating inflammatory bowel diseases |
| CN103142656A (en) * | 2013-03-18 | 2013-06-12 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparing composition for preventing and treating colon cancer |
| CN103404707A (en) * | 2013-08-19 | 2013-11-27 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparation of compositions for preventing intestinal diseases of animals |
| CN105274019A (en) * | 2014-07-08 | 2016-01-27 | 王军 | Liquid-solid combined fermentation process for Bacteroides fragilis |
| CN105434477B (en) * | 2015-10-29 | 2019-03-05 | 广州普维君健药业有限公司 | Application of the bacteroides fragilis in anti-aquatic pathogenic bacterium |
| CN109528775A (en) * | 2017-09-22 | 2019-03-29 | 中山大学 | Bacteroides fragilis is preparing the application in the drug for treating and preventing tumour |
| CN110025636B (en) * | 2018-01-12 | 2021-07-23 | 石家庄普维生物科技有限公司 | Application of bacteroides fragilis extract in preparation of composition for enhancing immunity |
| CN110283760A (en) * | 2019-07-30 | 2019-09-27 | 集美大学 | A kind of fermentation culture method of common eel source bacteroides fragilis |
| CN112587552B (en) * | 2020-09-17 | 2023-09-12 | 大连图腾生命科学发展有限公司 | Application of bacteroides fragilis839 in preparing medicament for treating or assisting in treating immune related diseases |
| CN115957243A (en) * | 2021-10-12 | 2023-04-14 | 广州知易生物科技有限公司 | Application of bacteroides fragilis in preventing and treating cancer-related diarrhea |
-
1990
- 1990-05-03 CN CN 90102847 patent/CN1029857C/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103082294A (en) * | 2013-01-21 | 2013-05-08 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparation of composition for treating diarrhea |
| CN103146620A (en) * | 2013-03-25 | 2013-06-12 | 广州知光生物科技有限公司 | Bacteroides fragilis with characteristics of probiotics |
| WO2019095508A1 (en) * | 2017-11-17 | 2019-05-23 | 中山大学 | Composition for enhancing immune function of t cells and preparation method therefor |
| CN113151083A (en) * | 2021-04-21 | 2021-07-23 | 中国疾病预防控制中心传染病预防控制所 | Bacteroides dorsalis probiotics and application thereof in preparation of influenza treatment or prevention medicine |
| CN113151083B (en) * | 2021-04-21 | 2022-05-20 | 中国疾病预防控制中心传染病预防控制所 | Bacteroides donovani probiotic and its application in the preparation of influenza therapeutic or preventive medicine |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1056314A (en) | 1991-11-20 |
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