Lactobacillus crispatus and its use

Abstract

The invention discloses Lactobacillus crispatus and its use. The Lactobacillus crispatus is Lactobacillus crispatus chen-01 (Lactobacillus crispatus chen-01), and its deposit number is CGMCC NO.23396. Lactobacillus crispatus can be used to prepare medicines, and the medicines are used to treat and / or prevent HPV infection and vaginal infection. After the medicine prepared by the Lactobacillus crispatus is locally transplanted into the vagina of the present invention, the HPV viral load can be effectively reduced, the HPV clearance rate can be significantly improved, vaginitis can be effectively improved, and the diversity and abundance of vaginal flora can be significantly reduced; in addition, local vaginal transplantation of Lactobacillus crispatus can also significantly improve the vaginal microecological composition, which is close to that of normal healthy people.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to Lactobacillus crispatus and uses thereof. Background Art

[0002] Cervical cancer is the most serious cervical lesion, a common gynecological malignancy, and the second leading cause of cancer death in women. It has been determined to be a solid tumor caused by human papilloma virus (HPV), and persistent infection with high-risk HPV is a necessary condition for cervical cancer and precancerous lesions. Human papillomavirus (HPV) is a small circular double-stranded DNA virus that infects the epidermis and mucosal squamous epithelium. According to the degree of HPV pathogenicity or the degree of carcinogenic risk, HPV can be divided into two categories: low-risk and high-risk. High-risk types include HPV16, 18, 31, 33, 35, 45, 51, 52, 56, 58, 61, etc. It has been reported that in invasive cervical cancer, intact HPV DNA is present in almost all cervical cancer biopsies. In invasive cervical cancer, intact HPV DNA is present in almost all cervical cancer biopsies. Its major HPV oncoproteins (especially E6 and E7) are involved in the transformation, immortalization and malignancy of cervical cancer.

[0003] Regarding the treatment of HPV infection, there is currently a lack of recognized effective treatment methods in clinical practice. The main methods include: physical therapy (mainly removing visible tumors and subclinical infections) including: laser, microwave, freezing, electrocautery, surgical resection (LEEP knife in gynecology, etc.); drug treatment: 0.5% podophyllotoxin tincture, 5% imiquimod, 50% trichloroacetic acid, fluorouracil ointment, etc.; immunotherapy: it aims to reduce recurrence and accelerate the removal of lesions, and the drugs include: interferon, interleukin, thymosin, transfer factor, BCG, isotretinoin, autologous vaccine, etc.; therapeutic vaccine: there is currently no effective therapeutic vaccine. However, all of the above methods have various defects, and the treatment effect is unsatisfactory. Therefore, finding a safe and effective anti-HPV treatment method has always been a research hotspot for the prevention and treatment of cervical precancerous lesions and cervical cancer.

[0004] Microecology is an emerging discipline. With the continuous development of society, microecology has attracted more and more attention from scholars. The vaginal microecology system is a more complex and important system among the four major ecosystems of the human body. It plays an important role in human reproduction and disease defense mechanism. The female vaginal microecology system is composed of anatomical structure, vaginal flora, local immune function and endocrine regulation function of the body. Among them, the normal flora in the vagina is the core content of the study of vaginal microecology system. As a complex ecosystem, the vaginal microecology changes with the age of the host and is in a dynamic equilibrium state during the menstrual cycle, pregnancy and different physical conditions. This balance is beneficial to the health of the host.

[0005] Under normal circumstances, dozens of microorganisms can be isolated from the female vagina, including conditional pathogens such as Candida albicans, Gardnerella, Escherichia coli, Enterococcus, Streptococcus, Staphylococcus epidermidis, etc. At the same time, probiotics such as Bifidobacterium and Lactobacillus can also be isolated. Pathogenic bacteria and probiotics are in a dynamic equilibrium state. The most common lactobacilli in the vagina are Lactobacillus crispatus, Lactobacillus inertus, Lactobacillus jensenii, Lactobacillus gasseri, etc. Vaginal lactobacilli are the dominant bacteria, living on the surface of the vaginal mucosa, metabolizing sugar to produce acid, making the vagina in an acidic environment, forming a biofilm with the mucus in the vagina, producing bacteriocins, bacteriocin-like substances and biosurfactants, preventing harmful bacteria from colonizing in the vagina, and activating a variety of immune cells to further promote the formation of host humoral immunity and cellular immunity, which is conducive to maintaining the balance of the vaginal microecology. When the balance is broken, in addition to causing various vaginal inflammations, it may even promote the occurrence of cancer.

[0006] In recent years, the role of changes in vaginal microecology in genital HPV infection has attracted the attention of more and more scholars. Through high-throughput sequencing of vaginal secretions of normal women with negative HPV and vaginal secretions of HPV-positive women, researchers found that there were obvious differences in vaginal microecology between the two. HPV-positive women mainly showed highly diverse vaginal microorganisms, a decrease in lactobacilli and an increase in vaginal Gardnerella. In addition, a large number of literatures have shown that bacterial vaginosis (BV) is positively correlated with the incidence, prevalence and persistence of HPV infection. BV is actually a kind of vaginal flora imbalance, the main phenotype of which is a decrease in lactobacilli that secrete H2O2 in the vagina, while Gardnerella and anaerobic bacteria multiply in large numbers. Under normal circumstances, lactobacilli prevent the colonization of bacterial species related to bacterial vaginosis by maintaining low pH and producing bacteriocins, which is important for maintaining the barrier function of the cervical epithelium that inhibits HPV from entering basal keratinocytes. When BV-associated strict anaerobes are able to colonize, they produce enzymes and metabolites that may damage this barrier, providing a favorable environment for HPV infection and invasion. The entry of HPV, especially high-risk HPV fragments and host gene integration and eventual tumor transformation. Therefore, HPV infection is inseparable from the disorder of the local microecological environment of the vagina, especially the reduction of Lactobacillus.

[0007] Probiotics are currently defined as “live microorganisms which, when administered in adequate amounts, confer a health benefit on the host.” Probiotics include live bacteria (such as Bifidobacteria, Lactobacilli, and Streptococci) that can modulate the composition of the microbiota to improve health, the immune system, inflammatory status, and bioavailability of micronutrients.

[0008] At present, probiotics have been widely used in gastrointestinal diseases (such as diarrhea and constipation, irritable bowel syndrome, various intestinal inflammatory diseases, eradication of Helicobacter pylori, etc.), as well as infant lactose intolerance, neonatal necrotizing enterocolitis, etc. It can reduce the risk of diabetes, improve dyslipidemia, treat cardiovascular diseases, allergic diseases and certain cancers, and has also been reported in preventing oral infections and dental caries, and even autism. In the female reproductive tract, probiotics have been proven to be a safe and effective method for the treatment of inflammatory diseases such as bacterial vaginosis and candidal vaginitis.

[0009] At present, there are relatively few literature reports on the clinical use of probiotics to treat cervical and vaginal HPV infection. Therefore, this study aims to explore the changes in the vaginal microecology of HPV-infected people and the therapeutic effect of probiotics on patients with cervical and vaginal HPV infection, in order to improve the clearance rate of HPV infection and provide a new method for the prevention and treatment of cervical intraepithelial neoplasia and cervical cancer. Summary of the invention

[0010] In view of the deficiencies and difficulties in the prior art, the present invention aims to provide a Lactobacillus crispatus and its use. After local vaginal transplantation of Lactobacillus crispatus, the vaginal microecology of HPV-infected people changes, which has a therapeutic effect on patients with cervical and vaginal HPV infection, improves the clearance rate of HPV infection, and provides a new method for preventing and treating cervical intraepithelial neoplasia and cervical cancer.

[0011] The present invention is achieved through the following technical solutions:

[0012] On the one hand, the present invention provides a Lactobacillus crispatus, which has been deposited in the General Microbiology Center of China Microorganism Culture Collection Committee (CGMCC for short) on September 13, 2021. The address of the depository unit is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, and the deposit registration number is CGMCC NO.23396. The strain is classified and named Lactobacillus crispatus chen-01.

[0013] Another aspect of the present invention provides use of the Lactobacillus crispatus in preparing a medicament for treating and / or preventing HPV infection.

[0014] Furthermore, the drug has the function of reducing HPV viral load and improving HPV clearance rate.

[0015] Another aspect of the present invention provides use of the above-mentioned Lactobacillus crispatus in preparing a medicine, characterized in that the medicine is used for treating and / or preventing vaginal infection.

[0016] Furthermore, the drug is used to significantly improve the vaginal microecology and reduce the diversity and abundance of vaginal flora.

[0017] The present invention also provides a pharmaceutical product, which contains the aforementioned Lactobacillus crispatus and optionally one or more pharmaceutically acceptable drug carriers and / or pharmaceutical excipients.

[0018] Compared with the prior art, the Lactobacillus crispatus chen-01 of the present invention is a completely new isolated strain. After local vaginal transplantation of the Lactobacillus crispatus, the HPV viral load can be effectively reduced, the HPV clearance rate can be significantly improved, vaginitis can be effectively improved, and the diversity and abundance of vaginal flora can be significantly reduced; in addition, local vaginal transplantation of the Lactobacillus crispatus can also significantly improve the vaginal microecological composition, which is close to that of normal healthy people. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Figure 3: Effects of intravaginal transplantation of L. crispatus chen-01 on the diversity and abundance of vaginal flora; Figure (A) compares Chao1, Observed species, Shannon, Simpson and Good's coverage of Hb, Hf and H-Bb groups; Figure (B) is the PCoA analysis before and after probiotic treatment; Figure (C) is a heat map of species composition before (Hb) and after (H-Bb) probiotic treatment.

[0020] Figure 2 The effects of intravaginal transplantation of L. crispatus chen-01 on the composition of vaginal microecology; Figure (A) is a comparison of the microecological composition at the vaginal phylum level among CH-b, Hf and H-Bb groups; Figure (B) is a comparison of the vaginal microecological composition at the genus level among C, Hb, Hf and H-Bb groups; Figures (C) to (E) are comparisons of the phylum composition of Firmicutes, Bacteroidetes and Actinobacteria among C, Hb, Hf and H-Bb groups, respectively; Figures (F) to (H) are comparisons of the genus composition of Lactobacillus, Prevotella and Gardnerella among C, Hb, Hf and H-Bb groups, respectively. DETAILED DESCRIPTION

[0021] The present invention is further described below in conjunction with the accompanying drawings. Unless otherwise specified, the reagents used in the experiments of the embodiments can be obtained from commercial sources.

[0022] Example 1 Screening and in vitro property evaluation of Lactobacillus crispatus chen-01

[0023] 1. Screening of Lactobacillus crispatus chen-01

[0024] Using the technology of culturing omics, the vaginal secretions of healthy women collected clinically were diluted in multiple proportions, plated for separation and screening, the DNA of the isolated colonies was extracted and sent to the company for sequencing, the strains were identified, and vaginal Lactobacillus crispatus was isolated and screened. On September 13, 2021, the Lactobacillus crispatus was deposited in the General Microbiology Center of the China Microbiological Culture Collection (CGMCC for short). The address of the deposit unit is: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, the deposit registration number is CGMCC NO.23396, and the strain is classified and named Lactobacillus crispatus chen-01 (Lactobacillus crispatus chen 01).

[0025] Screening bacteria identification sequencing sequence (SEQ ID NO.1):

[0026]

[0027] 2. In vitro evaluation of Lactobacillus crispatus chen-01

[0028] 1. Acid resistance test

[0029] Lactobacillus crispatus chen-01 was cultured in MRS medium at 37°C in a shaking incubator for 24 hours. After 24 hours, 100 μL of the bacterial solution was taken out and added into PBS buffers with pH=2, 4, 5, 6, and 7, respectively. After anaerobic culture for 4 hours, the plate was counted.

[0030] The results showed that Lactobacillus crispatus chen-01 had good tolerance to high concentration of acid.

[0031] 2. Antioxidant test

[0032] (1) Determination of DPPH free radical scavenging ability:

[0033] The Lactobacillus crispatus chen-01 strain was cultured in an MRS medium at 37° C. in a shaking incubator for 24 hours. After 24 hours, the bacterial solution was centrifuged at 8000 rpm for 3 minutes to obtain the supernatant.

[0034] Take 1 mL of bacterial supernatant and mix it with 1 mL of prepared DPPH free radical methanol solution. After shaking evenly, react at room temperature in the dark for 30 minutes, and measure its OD value at a wavelength of 517 nm (deionized water is used as a blank control).

[0035] DPPH free radical scavenging rate: [1-A517(sample) / A517(blank)]×100%

[0036] (2) Determination of the ability to scavenge hydroxyl free radicals:

[0037] The Lactobacillus crispatus chen-01 strain was cultured in MRS medium at 37° C. in a shaking incubator for 24 hours. After 24 hours, the bacterial solution was centrifuged at 8000 rpm for 3 minutes to obtain the supernatant.

[0038] Take 1mL of supernatant and add it to a glass test tube (sample tube), add 1mL of ddH2O to a blank tube, and add 1mL of 3mmol / L salicylic acid, 1mL of 1mmol / L FeSO4, and 1mL of 3mmol / L H2O2 respectively. After mixing evenly, react in a 37℃ water bath for 15min, and measure the absorbance at a wavelength of 510nm using a spectrophotometer.

[0039] Hydroxyl radical scavenging rate (%) = [1-A510 (sample) / A510 (blank)] × 100%

[0040] The results showed that Lactobacillus crispatus chen-01 exhibited good antioxidant capacity and DPPH, O2 and OH reducing ability.

[0041] 3. Antibacterial Experiment

[0042] The Lactobacillus crispatus chen-01 strain was cultured in MRS medium at 37° C. in a shaking incubator for 24 hours. After 24 hours, the bacterial solution was centrifuged at 8000 rpm for 3 minutes to obtain the supernatant.

[0043] Candida albicans, Staphylococcus aureus, Escherichia coli O157, beta-hemolytic Streptococcus, Enterococcus faecalis, and Gardnerella were spread on LB solid culture medium. The Oxford cup was gently placed on the plate, and 250 μL of the probiotic culture supernatant was drawn into the Oxford cup (two replicates were made for each group), and cultured at 37°C. The size of the inhibition zone was observed every 2 hours, and the diameter of the inhibition zone was measured after 8 hours.

[0044] The results showed that Lactobacillus crispatus chen-01 showed good inhibitory effect on Candida albicans, Staphylococcus aureus, Escherichia coli O157, beta-hemolytic Streptococcus, Enterococcus faecalis, and Gardnerella.

[0045] Example 2 Effect of local vaginal transplantation of Lactobacillus crispatus on women infected with high-risk HPV

[0046] 1. Description of research subjects

[0047] The study subjects were female patients with cervical HR-HPV infection recruited from the gynecology outpatient clinic or inpatient department of Nanchang First Hospital and Jingdezhen Maternal and Child Health Hospital from July 1, 2021 to June 30, 2022. By asking about the medical history and reviewing previous case data, 161 women with cervical HR-HPV infection were selected for inclusion in the study. All subjects underwent colposcopy and cervical biopsy when necessary. The age, maternal and childbirth history, smoking, contraceptive method, age of initial sexual behavior, number of sexual partners, etc. of the subjects were collected (Table 1). Informed consent was obtained from all patients. This project has been approved by the hospital medical ethics committee and registered on the official website of the China Clinical Trial Registry (registration number ChiCTR2100046239).

[0048] Inclusion criteria: ① The patient signed an informed consent form; ② Aged 18-65 years old, with a history of sexual life; ③ No use of any medication; ④ No sexual life, vaginal douching or medication 3 days before the examination; ⑤ No abnormal vaginal bleeding; ⑥ No concurrent systemic diseases: diabetes, diseases requiring hormone drugs (acute and chronic nephritis), etc.

[0049] Exclusion criteria: ① patients with high-grade cervical intraepithelial neoplasia or cervical cancer or reproductive tract tumors; ② women who do not agree to participate in the experiment; ③ subjects with a history of vaginal douching, medication, or sexual life 3 days before the examination; ④ subjects taking contraceptives, antibiotics, immunosuppressants and other drugs 3 days before the examination; ⑤ patients after total or subtotal hysterectomy; ⑥ patients with serious medical complications, such as diabetes, autoimmune diseases, immunodeficiency diseases, etc.; ⑦ patients allergic to probiotics.

[0050] Grouping and treatment: According to the inclusion and exclusion criteria, 100 people finally met the conditions. Using the random number table method, 100 research subjects were randomly assigned to the placebo group (Hf) and the probiotic group (H-Bb) at a ratio of 1:1. The probiotic preparation used in the H-Bb group of this embodiment is a mixture of Lactobacillus crispatus (L.crispatus chen-01, numbered CGMCC 23396) and sweet potato powder, and the colony forming unit (CFU) of each capsule is 1×109CFU (hereinafter referred to as probiotics). Patients in the probiotic group are given probiotic capsules locally in the vagina. The specific method of medication is: after cleaning the vulva, put on a finger cot, put this product into the deep part of the vagina, and apply the medicine before going to bed at night, one capsule each time, once a day. Use it every night for 10 consecutive days from the 1st to the 3rd month. Use it once every three days from the 4th to the 5th month, for a total of 5 times. Avoid the month period, and do not rinse the vagina during medication. Patients in the placebo group used capsules containing only the ingredients, and their usage was the same as that of the probiotic group.

[0051] At the beginning of the trial, 161 women participated. According to the inclusion and exclusion criteria, 100 eligible women were evaluated and finally randomly divided into a probiotic group (H-Bb group) and a placebo group (Hf group), with 50 cases in each group.

[0052] Table 1 Basic characteristics of patients (mean ± standard deviation)

[0053]

[0054]

[0055] During the study, 1 case withdrew from the H-Bb group and 2 cases were lost to follow-up; 2 cases withdrew from the Hf group and 2 cases were lost to follow-up. Finally, 45 cases in the probiotic group (H-Bb) and 46 cases in the placebo group (Hf group) completed the trial period.

[0056] 2. Specimen collection

[0057] Use a vaginal dilator to expose the cervix, wipe its secretions with a sterile cotton ball, and then rotate it clockwise 3 times with a special brush. Take out the brush and put it in a special preservation solution. One part of the sample is used for HPV DNA detection, and the other part is used for TCT detection of cytological classification. Hybrid capture II (HC II) detects HR-HPV types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, with HPV-DNA ≥ 1pg / mL as the positive standard. Real-time fluorescence quantitative PCR is used to detect HPV types. Cure is defined as all HPV subtypes turning negative. Effectiveness is defined as one or more HPV subtypes turning negative. Ineffectiveness is defined as an increase in HPV subtypes or no HPV subtype turning negative. HPV clearance rate (%) = cure rate (%) + effective rate (%). During TCT detection, the sample is rinsed to obtain cells, which are dispersed and filtered by an automatic cell detector, and then microscopically detected and analyzed.

[0058] 3. DNA extraction and high-throughput sequencing

[0059] In the sixth month after enrollment, vaginal secretion samples were collected 3 to 7 days after the menstruation ended. The patient was in the lithotomy position, and a gynecologist used a disposable vaginal speculum (the speculum was not coated with iodine or other disinfectants to avoid difficulty in distinguishing it from white blood cells under the microscope) to expose the cervix, and used a sterile dry cotton swab to collect an appropriate amount of vaginal secretions from the posterior fornix and 1 / 3 of the vaginal side wall of the patient, put it into a sterilized EP tube, sealed it, marked it, and placed it in a -80℃ refrigerator for use in 16S rDNA high-throughput sequencing.

[0060] 4. Local vaginal transplantation of L.criapatus chen-01 can effectively reduce HPV viral load

[0061] As shown in Table 2, the initial HPV viral loads of the probiotic group (H-Bb) and the placebo group (Hf) were 267.70±27.68 and 245.10±26.43, respectively, with no significant difference between the two groups (p=0.56>0.05). After probiotic treatment, the viral load of the patients was 113.10±21.69, which was significantly lower than that before treatment (p<0.01), and there was also a significant difference compared with the viral load of the control group patients after follow-up (p=0.01<0.05). However, there was no significant difference in the viral load of the placebo group patients after follow-up compared with the time of enrollment (p=0.27>0.05).

[0062] Table 2 HPV viral load in the two groups before and after treatment (mean ± standard deviation)

[0063]

[0064] 5. L. crispatus chen-01 vaginal transplantation can significantly improve HPV clearance rate

[0065] As shown in Table 3, after 6 months of follow-up, 21 cases in the probiotic group turned negative, 5 cases were effective, 19 cases were positive, and the total effective rate was 57.78%. In the placebo group (Hf group), 17 cases turned negative, 4 cases were effective, and 25 cases were positive, with a total effective rate of 45.65%. The total effective rate of the probiotic group was higher than that of the control group, but there was no significant difference between the two (p = 0.25 > 0.05).

[0066] Table 3 HPV clearance rate before and after treatment in the two groups (mean ± standard deviation)

[0067]

[0068] 6. Lactobacillus crispatus chen-01 vaginal transplantation can effectively improve vaginitis

[0069] As shown in Table 4, cervical liquid-based cytology analysis showed that compared with the time of enrollment, the improvement rate of cytological abnormalities in the Hf group was 34.62%, and the improvement rate of inflammation was 27.27%. The improvement rate of cytological abnormalities in the H-Bb group was 82.14%, and the improvement rate of vaginitis was 77.78%, both higher than that in the Hf group (p < 0.05). In addition, no adverse events occurred during this study.

[0070] Table 4 Changes of cytology and inflammation in the two groups before and after treatment (mean ± SD)

[0071]

[0072] 7. The diversity and abundance of vaginal flora were significantly reduced after local vaginal transplantation of L. crispatus chen-01

[0073] 16S RNA sequencing was used to analyze the effect of local transplantation of L. crispatus chen-01 on the diversity and abundance of vaginal flora. In order to more comprehensively evaluate the alpha diversity of the microbial community, this process uses Chao1 and Observedspecies indices to represent richness, Shannon and Simpson indices to represent diversity, and Good's coverage index to represent coverage. The results are shown in Figure 2. Figure 1 As shown, Hb: HR-HPV patients before enrollment; Hf: HR-HPV patients in the placebo group; H-Bb: HR-HPV patients after probiotic treatment. Figure 1As shown in A, the Chao1, Observed species, Shannon and Simpson indexes of patients receiving probiotic treatment were significantly reduced (p < 0.05) compared with those before treatment, while the Goodscoverage index was significantly increased (p < 0.05). Beta diversity analysis was performed using PCoA. Figure 1 As shown in B, the probiotic group showed obvious aggregation after treatment. In order to further compare the differences in species composition between samples and to display the species abundance distribution trend of each sample, a heat map was used for species composition analysis. Figure 1 As shown in C, Lactobacillus increased significantly after probiotic treatment compared with before treatment.

[0074] 8. Local vaginal transplantation of Lactobacillus crispatus significantly improves the composition of vaginal microecology

[0075] The vaginal microecological composition of each group of patients was analyzed from the perspective of phylum and genus. The effect of intravaginal transplantation of L. crispatus chen-01 on the vaginal microecological composition was as follows: Figure 2 As shown, in the figure, C: healthy people without HPV infection; Hb: HR-HPV patients before enrollment; Hf: HR-HPV patients treated with placebo; H-Bb: HR-HPV patients after probiotic treatment. *p<0.05; **p<0.01.

[0076] At door level (such as Figure 2 A), in the normal healthy population (Group C), the vaginal flora was dominated by Firmicutes (95.13%), and the remaining bacteria accounted for less than 5%. Before HR-HPV treatment, Firmicutes accounted for (60.1%), and the remaining main bacteria were Bacteroidetes (21.20%), Fusobacteria (11.93%), and Actinobacteria (3.6%). Compared with the two, the HR-HPV group showed obvious flora diversity (p = 0.0058). After probiotic treatment in the HR-HPV group, the vaginal flora was close to that of the normal population, showing Firmicutes as the dominant bacteria (94.96%). Compared with before treatment, the flora diversity was significantly different (p = 0.0009), while there was no statistical significance in the control group (p = 0.0789).

[0077] Further analysis at the genus level (e.g. Figure 2B), in the normal healthy population (Group C), Lactobacillus was the dominant vaginal flora accounting for 94.74%, and also included Prevotella, Sneathia, and Gardnerella, but the content was very small. Before HR-HPV treatment, Lactobacillus accounted for 56.45%, and the abundance of the other main bacteria Prevotella (13.51%), Sneathia (6.42%), and Gardnerella (3.17%) increased significantly. Compared with the healthy population, the vaginal microecology showed obvious flora diversity, but after probiotic treatment, the diversity of vaginal flora also decreased, close to that of the normal healthy population (Lactobacillus accounted for 90.75%).

[0078] Figure 2 C-2H specifically shows the distribution of Firmicutes, Bacteroidetes, Actinobacteria, Lactobacillus, Prevotella and Gardnerella in the two groups of patients.

[0079] The above only expresses the preferred embodiments of the present invention, and the description is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that for ordinary technicians in this field, several modifications, improvements and substitutions can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the attached claims.

Claims

1. Lactobacillus crispatus, characterized in that: The Lactobacillus crispatus is Lactobacillus crispatus chen-01 ( Lactobacilluscrispatus chen-01), and its deposit number is CGMCC NO.23396.

2. The use of Lactobacillus crispatus according to claim 1 in preparing medicines, characterized in that: The medicament is used to treat HPV infection.

3. The use according to claim 2, characterized in that: The drug has the function of reducing HPV viral load.

4. The use according to claim 2, characterized in that: The drug has the function of improving the HPV clearance rate.

5. The use of Lactobacillus crispatus according to claim 1 in preparing medicines, characterized in that: The medicine is used for treating vaginitis.

6. The use according to claim 5, characterized in that: The drug is used to reduce the diversity and abundance of vaginal flora.

7. A pharmaceutical product comprising the Lactobacillus crispatus according to claim 1 and optionally one or more pharmaceutically acceptable drug carriers and / or pharmaceutical excipients.

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