CN103911307B - Lactobacillus crispatus bacterial strain and uses thereof - Google Patents
Lactobacillus crispatus bacterial strain and uses thereof Download PDFInfo
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- CN103911307B CN103911307B CN201310004353.4A CN201310004353A CN103911307B CN 103911307 B CN103911307 B CN 103911307B CN 201310004353 A CN201310004353 A CN 201310004353A CN 103911307 B CN103911307 B CN 103911307B
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- NIPNSKYNPDTRPC-UHFFFAOYSA-N N-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 NIPNSKYNPDTRPC-UHFFFAOYSA-N 0.000 description 1
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- GSXRBRIWJGAPDU-BBVRJQLQSA-N tyrocidine A Chemical compound C([C@H]1C(=O)N[C@H](C(=O)N[C@@H](CCCN)C(=O)N[C@H](C(N[C@H](CC=2C=CC=CC=2)C(=O)N2CCC[C@H]2C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N1)=O)CC(C)C)C(C)C)C1=CC=C(O)C=C1 GSXRBRIWJGAPDU-BBVRJQLQSA-N 0.000 description 1
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Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a kind of bacterial strain, does is this bacterial strain lactobacillus crispatus (Lactobacillus? crispatus) bacterial strain, is characterized in that, does is the preserving number of this bacterial strain CGMCC? No.6363.Present invention also offers above-mentioned bacterial strains for the preparation of the purposes prevented and/or treated in colpitic medicine, in the purposes prepared in vaginal care articles for use with preparing the purposes in genitalia sanitary product.By technique scheme, the present invention can realize the antagonistic action to various pathogens effectively.In addition, bacterial strain of the present invention also has the higher adhesive capacity to vaginal epithelial cell.Thus bacterial strain of the present invention can meet actual clinical application demand preferably.
Description
Technical field
The present invention relates to Medical Microbiology technical field, particularly, relate to a kind of bacterial strain of lactobacillus crispatus and the purposes of this bacterial strain.
Background technology
There are some dominant microfloras in healthy women vagina, these dominant microfloras play a part key to the balance maintaining vagina normal microflora.Dominant microflora plays by materials such as secretion lactic acid, hydrogen peroxide, bacteriocins the effect killing pathogenic bacterium.Meanwhile, play the effect of biological barrier at the competition adhesion of vaginal mucosal epithelium cell, prevent pathogenic bacterium in the breeding of vagina.
But many reasons such as antibiotic use, bad health habit, environment, chemicotherapy, stress can destroy vagina normal microflora balance, dominant microflora quantity is reduced, pathogenic bacterium amount reproduction, and cause various vaginopathy as bacterial vaginitis, monilial vaginitis, trichomonal vaginitis, child's property vaginitis, menstruation vaginitis, senile vaginitis, mixed infective vaginitis etc.
Recover Lactobacillus vaginalis advantage by the intervention of external source bacterium, thus the equilibrium state reaching normal vagina flora is a kind of effective means preventing and/or treating colpitis.But known external source bacterium is still more weak to the antagonistic action of pathogenic bacterium, be difficult to meet actual clinical application demand.
Summary of the invention
The object of this invention is to provide a kind of external source bacteria strain stronger to the antagonistic action of pathogenic bacterium, this bacterial strain can meet actual clinical application demand preferably.
To achieve these goals, the present inventor is separated and obtains a strain Bacterium lacticum from the vagina of volunteer, finds that this lactobacterium strain has the stronger antagonistic action to pathogenic bacterium unexpectedly, resulting in the present invention.
On the one hand, the invention provides a kind of bacterial strain, this bacterial strain is the bacterial strain of lactobacillus crispatus (Lactobacilluscrispatus), it is characterized in that, the preserving number of this bacterial strain is CGMCCNo.6363.
On the other hand, present invention also offers above-mentioned bacterial strains for the preparation of prevention colpitic medicine in purposes.
On the other hand, present invention also offers above-mentioned bacterial strains for the preparation for the treatment of colpitic medicine in purposes.
On the other hand, present invention also offers above-mentioned bacterial strains and prepare the purposes in vaginal care articles for use.
On the other hand, present invention also offers above-mentioned bacterial strains and prepare the purposes in genitalia sanitary product.
By technique scheme, the present invention can realize the antagonistic action to various pathogens effectively.In addition, bacterial strain of the present invention also has the higher adhesive capacity to vaginal epithelial cell and higher lactic acid producing ability.Thus bacterial strain of the present invention can meet actual clinical application demand preferably.
Other features and advantages of the present invention are described in detail in embodiment part subsequently.
Biological deposits
Lactobacillus crispatus of the present invention (Lactobacilluscrispatus) bacterial strain, (address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms's common micro-organisms center is deposited on July 17th, 2012, Institute of Microorganism, Academia Sinica, postcode: 100101) (depositary institution is abbreviated as CGMCC), deposit number is CGMCCNo.6363.
Embodiment
Below the specific embodiment of the present invention is described in detail.Should be understood that, embodiment described herein, only for instruction and explanation of the present invention, is not limited to the present invention.
On the one hand, the invention provides a kind of bacterial strain, this bacterial strain is the bacterial strain of lactobacillus crispatus (Lactobacilluscrispatus), it is characterized in that, the preserving number of this bacterial strain is CGMCCNo.6363.
Wherein, this bacterial strain has the 16SrDNA sequence shown in SEQIDNO:1.16SrDNA sequence is the DNA sequence dna of coding 16SrRNA, and the classification and/or qualification of bacterial strain have important reference value.
Wherein, the colonial morphology of this bacterial strain on MRS solid medium is circular colonies, diameter 0.5-0.75mm, surface irregularity, under the microscope in Gram-positive tyrothricin.
Wherein, the culture condition of this bacterial strain can adopt the cultural method of known lactobacillus crispatus, such as, this bacterial strain can on MRS liquid nutrient medium and MRS solid medium normal growth.Wherein, the growth temperature of this bacterial strain can be conventional temperature, and such as, growth temperature can be 36-38 DEG C.
In the present invention, MRS liquid nutrient medium refers to the MgSO of tween-80 by the ammonium citrate of the sodium acetate of the glucose of the extractum carnis of the yeast extract paste of the peptone of 10g, 5g, 10g, 20g, 5g, 2g, 1mL, 0.58g
47H
2the MnSO of O, 0.25g
44H
2the K of O, 2g
2hPO
4mix with the distilled water of 1000mL, adjust ph is 6.2-6.4 and the substratum obtained after 121 DEG C of high pressure steam sterilizations.
In the present invention, MRS solid medium refers to the MgSO of tween-80 by the ammonium citrate of the sodium acetate of the glucose of the extractum carnis of the yeast extract paste of the peptone of 10g, 5g, 10g, 20g, 5g, 2g, 1mL, 0.58g
47H
2the MnSO of O, 0.25g
44H
2the K of O, 2g
2hPO
4, the agar powder of 20g and 1000mL distilled water mixing, adjust ph is 6.2-6.4 and the substratum obtained after 121 DEG C of high pressure steam sterilizations.
On the other hand, present invention also offers above-mentioned bacterial strains for the preparation of prevention colpitic medicine in purposes.
Wherein, described vaginitis can be at least one in bacterial vaginitis, monilial vaginitis, trichomonal vaginitis, child's property vaginitis, menstruation vaginitis, senile vaginitis and mixed infective vaginitis.
Wherein, described vaginitis can be the vaginitis that at least one in Candida albicans (Candidaalbicans), intestinal bacteria (Escherichiacoli) and streptococcus aureus (Staphylococcusaureus) causes.
Wherein, above-mentioned bacterial strains can play the colpitic effect of prevention in the following way: (1) inoculates above-mentioned bacterial strains in MRS liquid nutrient medium, and cultivates as cell concentration is 10
7individual thalline/every mL, namely obtains the nutrient solution of above-mentioned bacterial strains; (2) above-mentioned nutrient solution is dipped with Sterilized sanitation cotton balls, wiping vagina, every day 1-2 time, each nutrient solution using 3-10mL.That is, namely the nutrient solution of above-mentioned bacterial strains can be used as preventing colpitic medicine.
On the other hand, present invention also offers above-mentioned bacterial strains for the preparation for the treatment of colpitic medicine in purposes.
Wherein, described vaginitis is at least one in bacterial vaginitis, monilial vaginitis, trichomonal vaginitis, child's property vaginitis, menstruation vaginitis, senile vaginitis and mixed infective vaginitis.
Wherein, described vaginitis can be the vaginitis that at least one in Candida albicans (Candidaalbicans), intestinal bacteria (Escherichiacoli) and streptococcus aureus (Staphylococcusaureus) causes.
Wherein, above-mentioned bacterial strains can play the colpitic effect for the treatment of in the following way: (1) inoculates above-mentioned bacterial strains in MRS liquid nutrient medium, and cultivates as cell concentration is 10
7individual thalline/every mL, namely obtains the nutrient solution of above-mentioned bacterial strains; (2) above-mentioned nutrient solution is dipped with Sterilized sanitation cotton balls, wiping vagina, every day 1-2 time, each nutrient solution using 3-10mL.That is, namely the nutrient solution of above-mentioned bacterial strains can be used as and is used for the treatment of colpitic medicine.
On the other hand, present invention also offers above-mentioned bacterial strains and prepare the purposes in vaginal care articles for use.
On the other hand, present invention also offers above-mentioned bacterial strains and prepare the purposes in genitalia sanitary product.
Below will be described the present invention by embodiment.
Testing example 1
According to the document (foundation of the sequence measurement of polluted bacteria in 16SrDNA Rapid identification blood products, Ying Yanling etc., China's microbiology and Journal of Immunology, 10 phases in 2009) in method, check order to the sequence of the 16SrDNA of lactobacillus crispatus bacterial strain CGMCCNo.6363 of the present invention, sequencing result confirms to have the 16SrDNA sequence shown in SEQIDNO:1 to lactobacillus crispatus bacterial strain of the present invention.
Embodiment 1
The present embodiment is for illustration of the antagonistic action of bacterial strain of the present invention to Candida albicans (Candidaalbicans).Candida albicans used in the present embodiment is be the bacterial strain of 10231 purchased from the article number of ATCC.
Being cultivated by lactobacillus crispatus CGMCCNo.6363 MRS liquid nutrient medium is cell concentration 1 × 10
7the nutrient solution of CFU/mL, called after nutrient solution A.
Being cultivated with MRS liquid nutrient medium by Candida albicans (Candidaalbicans) is cell concentration 1 × 10
7the nutrient solution of CFU/mL, called after nutrient solution B.
Above-mentioned nutrient solution A and above-mentioned nutrient solution B equal-volume are mixed as experimental group, by independent above-mentioned nutrient solution B as a control group.
Respectively by the experimental group of 1mL and control group nutrient solution, be inoculated in the MRS liquid nutrient medium of 150mL, cultivate 24 hours at 37 DEG C, obtain the nutrient solution of Dual culture.
Get the Dual culture nutrient solution 100 μ L of experimental group and control group respectively, be inoculated into (Sabouraud glucose agar in 15mL Candida albicans selective medium flat board, purchased from Qingdao high-tech park Hai Bo Bioisystech Co., Ltd, article No.: HB0235-5) in, select cultivation at 37 DEG C 24 hours, then Candida albicans is counted.
Count results shows, and the Candida albicans cell concentration of experimental group is only 4.67 × 10
5cFU/mL, and the Candida albicans cell concentration of control group is up to 4.67 × 10
6cFU/mL.
Comparative example 1
Test according to the method for embodiment 1, unlike, by the lactobacillus crispatus CGMCCNo.6363 in embodiment 1, the article number replaced with respectively purchased from ATCC is the lactobacillus crispatus of 33820.
Count results shows, and be the lactobacillus crispatus of 33820 for the article number purchased from ATCC, the Candida albicans cell concentration of experimental group also reaches 1.95 × 10
6cFU/mL, and the Candida albicans cell concentration of control group is still 4.67 × 10
6cFU/mL.
Embodiment 2
The present embodiment is for illustration of the antagonistic action of bacterial strain of the present invention to intestinal bacteria (Escherichiacoli).Intestinal bacteria used in the present embodiment are be the bacterial strain of 35218 purchased from the article number of ATCC.
Being cultivated by lactobacillus crispatus CGMCCNo.6363 MRS liquid nutrient medium is cell concentration 1 × 10
7the nutrient solution of CFU/mL, called after nutrient solution A.
Being cultivated with MRS liquid nutrient medium by intestinal bacteria (Escherichiacoli) is cell concentration 1 × 10
7the nutrient solution of CFU/mL, called after nutrient solution B.
Above-mentioned nutrient solution A and above-mentioned nutrient solution B equal-volume are mixed as experimental group, by independent above-mentioned nutrient solution B as a control group.
Respectively by the experimental group of 1mL and control group nutrient solution, be inoculated in the MRS liquid nutrient medium of 150mL, cultivate 24 hours at 37 DEG C, obtain the nutrient solution of Dual culture.
Get the Dual culture nutrient solution 100 μ L of experimental group and control group respectively, be inoculated into (eosin methylene blue nutrient agar in 15mL E. coli selectable culture medium flat plate, purchased from Qingdao high-tech park Hai Bo Bioisystech Co., Ltd, article No.: HB7010) in, select cultivation at 37 DEG C 24 hours, then intestinal bacteria are counted.
Count results shows, and the coli somatic concentration of experimental group is only 0CFU/mL, that is, do not find coli somatic, and the coli somatic concentration of control group is up to 2.04 × 10
8cFU/mL.
Comparative example 2
Test according to the method for embodiment 2, unlike, by the lactobacillus crispatus CGMCCNo.6363 in embodiment 1, the article number replaced with respectively purchased from ATCC is the lactobacillus crispatus of 33820.
Count results shows, and be the lactobacillus crispatus of 33820 for the article number purchased from ATCC, the coli somatic concentration of experimental group also reaches 7.76 × 10
7cFU/mL, and the coli somatic concentration of control group is still 2.04 × 10
8cFU/mL.
Embodiment 3
The present embodiment is for illustration of the antagonistic action of bacterial strain of the present invention to streptococcus aureus (Staphylococcusaureus).Streptococcus aureus used in the present embodiment is be the bacterial strain of 25923 purchased from the article number of ATCC.
Being cultivated by lactobacillus crispatus CGMCCNo.6363 MRS liquid nutrient medium is cell concentration 1 × 10
7the nutrient solution of CFU/mL, called after nutrient solution A.
Being cultivated with MRS liquid nutrient medium by streptococcus aureus (Staphylococcusaureus) is cell concentration 1 × 10
7the nutrient solution of CFU/mL, called after nutrient solution B.
Above-mentioned nutrient solution A and above-mentioned nutrient solution B equal-volume are mixed as experimental group, by independent above-mentioned nutrient solution B as a control group.
Respectively by the experimental group of 1mL and control group nutrient solution, be inoculated in the MRS liquid nutrient medium of 150mL, cultivate 24 hours at 37 DEG C, obtain the nutrient solution of Dual culture.
Get the Dual culture nutrient solution 100 μ L of experimental group and control group respectively, be inoculated into (N.F,USP MANNITOL sodium-chlor nutrient agar on the streptococcus aureus selective medium flat board of 15L, purchased from Qingdao high-tech park Hai Bo Bioisystech Co., Ltd, article No.: HB4128-1) in, select cultivation at 37 DEG C 24 hours, then streptococcus aureus is counted.
Count results shows, and the streptococcus aureus cell concentration of experimental group is only 0CFU/mL, that is, do not find streptococcus aureus thalline, and the streptococcus aureus cell concentration of control group is up to 7.94 × 10
7cFU/mL.
Comparative example 3
Test according to the method for embodiment 1, unlike, by the lactobacillus crispatus CGMCCNo.6363 in embodiment 1, the article number replaced with respectively purchased from ATCC is the lactobacillus crispatus of 33820.
Count results shows, and the article number for ATCC is the lactobacillus crispatus of 33820, and the streptococcus aureus cell concentration of experimental group also reaches 3.89 × 10
7cFU/mL, and the streptococcus aureus cell concentration of control group is still 7.94 × 10
7cFU/mL.
Result according to embodiment 1-3 and comparative example 1-3 can be found out, lactobacillus crispatus CGMCCNo.6363 of the present invention has excellent antagonistic action to Candida albicans (Candidaalbicans), intestinal bacteria (Escherichiacoli) and streptococcus aureus (Staphylococcusaureus).
Embodiment 4
The present embodiment is according to document (Wang Hongyan, Lactobacterium acidophilum is sticked and the research of Antagonism effect reproductive tract is epithelial, China's microbiology and Journal of Immunology, volume the 10th phase October the 24th in 2004) in method measure lactobacillus crispatus CGMCCNo.6363 of the present invention to the adhesive capacity of vaginal epithelial cell.
After measured, lactobacillus crispatus CGMCCNo.6363 of the present invention is 16.3085 ± 1.74 to the adhesion index of vaginal epithelial cell.
Comparative example 4
The article number measured purchased from ATCC according to the method for embodiment 4 is the lactobacillus crispatus of 33820.
After measured, the lactobacillus crispatus being 33820 purchased from the article number of ATCC is 10.37 ± 1.85 to the adhesion index of vaginal epithelial cell.
Result according to embodiment 4 and comparative example 4 can be found out, lactobacillus crispatus CGMCCNo.6363 of the present invention has the higher adhesive capacity to vaginal epithelial cell.
Embodiment 5
The present embodiment is according to document (Fan Yonghong, Wang Li, Liu Dan, etc. the study on determination method [J] of Rhizopus oryzae Lactic Acid from Fermentation Broth content. biotechnology, 2007,17(1): the method 54-55) measures the lactic acid content of lactobacillus crispatus CGMCCNo.6363 of the present invention.
After measured, the lactic acid content of lactobacillus crispatus CGMCCNo.6363 of the present invention is 6.7684g/L.
Comparative example 5
Measuring purchased from the article number of ATCC according to the method for embodiment 5 is the lactic acid content of the lactobacillus crispatus of 33820.
After measured, the lactic acid content being the lactobacillus crispatus of 33820 purchased from the article number of ATCC is 6.7834g/L.
Result according to embodiment 5 and comparative example 5 can be found out, lactobacillus crispatus CGMCCNo.6363 of the present invention has higher lactic acid producing ability.
More than describe the preferred embodiment of the present invention in detail; but the present invention is not limited to the detail in above-mentioned embodiment, within the scope of technical conceive of the present invention; can carry out multiple simple variant to technical scheme of the present invention, these simple variant all belong to protection scope of the present invention.
It should be noted that in addition, each concrete technical characteristic described in above-mentioned embodiment, in reconcilable situation, can be combined by any suitable mode, in order to avoid unnecessary repetition, the present invention illustrates no longer separately to various possible array mode.
In addition, also can carry out arbitrary combination between various different embodiment of the present invention, as long as it is without prejudice to thought of the present invention, it should be considered as content disclosed in this invention equally.
Claims (10)
1. a bacterial strain, this bacterial strain is the bacterial strain of lactobacillus crispatus (Lactobacilluscrispatus), it is characterized in that, the preserving number of this bacterial strain is CGMCCNo.6363.
2. bacterial strain according to claim 1, wherein, this bacterial strain has the 16SrDNA sequence shown in SEQIDNO:1.
3. the bacterial strain described in claim 1 or 2 for the preparation of prevention colpitic medicine in purposes.
4. purposes according to claim 3, wherein, described vaginitis is at least one in bacterial vaginitis, monilial vaginitis, trichomonal vaginitis, child's property vaginitis, menstruation vaginitis, senile vaginitis and mixed infective vaginitis.
5. purposes according to claim 3, wherein, described vaginitis is the vaginitis that at least one in Candida albicans (Candidaalbicans), intestinal bacteria (Escherichiacoli) and streptococcus aureus (Staphylococcusaureus) causes.
6. the bacterial strain described in claim 1 or 2 for the preparation for the treatment of colpitic medicine in purposes.
7. purposes according to claim 6, wherein, described vaginitis is at least one in bacterial vaginitis, monilial vaginitis, trichomonal vaginitis, child's property vaginitis, menstruation vaginitis, senile vaginitis and mixed infective vaginitis.
8. purposes according to claim 6, wherein, described vaginitis is the vaginitis that at least one in Candida albicans (Candidaalbicans), intestinal bacteria (Escherichiacoli) and streptococcus aureus (Staphylococcusaureus) causes.
9. the bacterial strain described in claim 1 or 2 is preparing the purposes in vaginal care articles for use.
10. the bacterial strain described in claim 1 or 2 is preparing the purposes in genitalia sanitary product.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310004353.4A CN103911307B (en) | 2013-01-05 | 2013-01-05 | Lactobacillus crispatus bacterial strain and uses thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310004353.4A CN103911307B (en) | 2013-01-05 | 2013-01-05 | Lactobacillus crispatus bacterial strain and uses thereof |
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| Publication Number | Publication Date |
|---|---|
| CN103911307A CN103911307A (en) | 2014-07-09 |
| CN103911307B true CN103911307B (en) | 2016-04-13 |
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| CN107937324B (en) * | 2018-01-10 | 2020-01-21 | 中国科学院微生物研究所 | Lactobacillus crispatus and application thereof |
| CN109394796A (en) * | 2018-12-06 | 2019-03-01 | 青岛东海药业有限公司 | Lactobacillus crispatus preparation and its application |
| CN114480198A (en) * | 2022-02-11 | 2022-05-13 | 西南大学 | Lactobacillus crispatus strain for vagina and application thereof |
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| CN1556852A (en) * | 2001-09-20 | 2004-12-22 | �����ɷ� | Lactic acid producing bacteria for use as probiotic organisms in the human vagina |
| CN101048168A (en) * | 2004-10-05 | 2007-10-03 | 普罗比公司 | Probiotic lactobacillus strains for improved vaginal health |
| CN102727531A (en) * | 2012-07-23 | 2012-10-17 | 郑州金森生物科技工程有限公司 | Active lactic acid bacteria capsules with treatment and prevention effect on vaginitis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1556852A (en) * | 2001-09-20 | 2004-12-22 | �����ɷ� | Lactic acid producing bacteria for use as probiotic organisms in the human vagina |
| CN101048168A (en) * | 2004-10-05 | 2007-10-03 | 普罗比公司 | Probiotic lactobacillus strains for improved vaginal health |
| CN102727531A (en) * | 2012-07-23 | 2012-10-17 | 郑州金森生物科技工程有限公司 | Active lactic acid bacteria capsules with treatment and prevention effect on vaginitis |
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