Intrauterine anti-infection and anti-adhesion gel and preparation method thereof

Abstract

The invention discloses an intrauterine anti-infection and anti-adhesion gel and a preparation method thereof, the intrauterine anti-infection and anti-adhesion gel comprises lactobacillus rhamnosus CGMCC 1.577 and lactococcus lactis CGMCC 1.15072, the fermentation product of lactobacillus rhamnosus CGMCC 1.577 has uniqueness, can generate a synergistic effect with the lactococcus lactis CGMCC 1.15072, and can have relatively excellent effects of reducing endometritis cell infiltration, interstitial and epithelial reactive changes, improving the anti-infection and anti-adhesion effects and improving the anti-infection and anti-adhesion effects. The number of infectious microbes is reduced, inflammatory factors are reduced, and the endometritis disease can be well improved.

Description

TECHNICAL FIELD

[0001] The present application relates to a female health and hygiene product, in particular to an intrauterine anti-infection anti-adhesion gel and a preparation method thereof. BACKGROUND

[0002] Endometritis, as a common gynecological disease, affects the health of a large number of women worldwide. According to statistics, the incidence of endometritis in sexually active women of childbearing age shows an increasing trend year by year, which has a serious impact on the reproductive health and quality of life of patients. If not treated in time and effectively, endometritis not only can cause serious complications such as infertility and ectopic pregnancy, but also can lead to problems such as intrauterine adhesion, chronic pelvic pain, and intrauterine abscess, further aggravating the patient's suffering, and also bringing heavy economic burden to the family and society.

[0003] At present, the treatment methods for endometritis in clinic mainly include systemic therapy and local therapy. Systemic therapy usually uses intravenous infusion, intramuscular injection or oral administration of antibacterial drugs, however, due to the special physiological structure of the uterus, the drugs are difficult to effectively reach the affected area, resulting in unsatisfactory efficacy. Local therapy usually uses antibacterial drugs or disinfectant solutions to flush the uterus, which can kill pathogenic bacteria and remove inflammatory secretions to some extent, but with the widespread use of antibacterial drugs, drug-resistant strains are increasing, and the treatment effect is gradually declining. In addition, the drug maintenance time of solution flushing is short, in order to achieve better treatment effect, frequent flushing is required, which not only increases the work intensity of medical staff, but also brings many inconveniences to patients.

[0004] Among the many drug forms for treating endometritis, gels have received widespread attention due to their unique advantages. Gels have good adhesion and can stay in the uterus for a long time, prolonging the contact time of drugs with the affected area, thereby improving the efficacy of drugs. However, the existing gels for treating endometritis have unsatisfactory effects.

[0005] In order to improve the treatment effect, it is of great clinical significance and practical demand to develop an intrauterine gel for treating endometritis, which is efficient, stable and has good antibacterial properties.

[0006] The present application aims to provide an intrauterine anti-infection anti-adhesion gel and a preparation method thereof, which is expected to provide new treatment options for a large number of patients with endometritis and potential postoperative endometritis, make up for the shortcomings of existing treatment methods, and has great clinical significance and market value. SUMMARY

[0007] The application aims to provide a preparation method of an anti-infection anti-adhesion gel for intrauterine use, and the obtained anti-infection gel for intrauterine use can effectively improve the intrauterine inflammatory state, reduce the number of intrauterine bacteria, increase the endometrial thickness, and reduce intrauterine adhesion, thereby treating endometritis.

[0008] The bacteria used in the patent are all commercially available.

[0009] The Lactobacillus rhamnosus is deposited in the China General Microbiological Culture Collection Center, and the deposit number is CGMCC 1.577.

[0010] The Lactobacillus rhamnosus is deposited in the China General Microbiological Culture Collection Center, and the deposit number is CGMCC 1.484.

[0011] The Lactococcus lactis is deposited in the China General Microbiological Culture Collection Center, and the deposit number is CGMCC 1.15072.

[0012] The Lactococcus lactis is deposited in the China General Microbiological Culture Collection Center, and the deposit number is CGMCC 1.62.

[0013] During the research and practice, the research team of the applicant found that the fermentation product of Lactobacillus rhamnosus CGMCC 1.577 has its unique properties, can produce a synergistic effect with Lactococcus lactis CGMCC 1.15072, can have excellent reduction of intrauterine inflammatory cell infiltration, interstitial and epithelial reactivity, reduction of the number of miscellaneous bacteria, reduction of inflammatory factors, and can better improve the endometritis symptoms.

[0014] A composition of an anti-infection gel for intrauterine use:

[0015] The raw materials of the gel are, by mass percentage, 3% of Lactobacillus rhamnosus supernatant, 3% of Lactococcus lactis supernatant, 25% of sophora flavescens essential oil, 25% of hyaluronic acid, 4% of water-soluble vitamin E nicotinic acid ester, 2.5% of vitamin E, 2.5% of carboxymethyl cellulose sodium, and 35% of water.

[0016] Preparation of the bacterial supernatant:

[0017] Lactobacillus rhamnosus CGMCC 1.577 strain was thawed, inoculated on MRS solid culture medium, and cultured in a 37 DEG C incubator for 24 hours, and single colonies with good morphology were picked and inoculated in MRS liquid medium for activation, and cultured at 37 DEG C, 150r / min for 24 hours, then centrifuged at 5000xg for 10 minutes, and the supernatant was filtered with a 0.22 mu bacteria filter to obtain Lactobacillus rhamnosus supernatant, which was stored at 4 DEG C for later use.

[0018] Lactococcus lactis CGMCC 1.15072 strain was thawed, inoculated on MRS solid culture medium, and cultured in a 30 DEG C incubator for 24 hours, and single colonies with good morphology were picked and inoculated in MRS liquid medium for activation, and cultured at 30 DEG C, 200r / min for 24 hours, then centrifuged at 5000xg for 10 minutes, and the supernatant was filtered with a 0.22 mu bacteria filter to obtain Lactococcus lactis supernatant, which was stored at 4 DEG C for later use.

[0019] Preparation of anti-infection gel:

[0020] Water was heated to 30 DEG C, and water-soluble vitamin E nicotinate and vitamin E were added while stirring, and then the temperature was raised to 40 DEG C after stirring for 30 minutes, and then Lactobacillus rhamnosus supernatant, Lactococcus lactis supernatant, Sophora flavescens essential oil, hyaluronic acid and sodium carboxymethyl cellulose were added and stirred for 2 hours to obtain an anti-infection gel for intrauterine use.

[0021] Advantages of the present application:

[0022] (1) Lactobacillus rhamnosus CGMCC 1.577 fermentation product has its unique nature, can produce a synergistic effect with Lactococcus lactis CGMCC 1.15072, can have excellent reduction of intrauterine inflammatory cell infiltration, interstitial and epithelial reactivity, reduction of the number of miscellaneous bacteria, reduction of inflammatory factors, and can better improve endometritis symptoms. DETAILED DESCRIPTION

[0023] The embodiments of the present application are described in detail below, which are only used to explain the present application and cannot be understood as a limitation of the present application.

[0024] Example 1

[0025] A composition of an anti-infection anti-adhesion gel for intrauterine use:

[0026] The raw materials of the gel are, by mass percentage, Lactobacillus rhamnosus supernatant 3%, Lactococcus lactis supernatant 3%, Sophora flavescens essential oil 25%, hyaluronic acid 25%, water-soluble vitamin E nicotinate 4%, vitamin E 2.5%, sodium carboxymethyl cellulose 2.5%, and water 35%.

[0027] Preparation of the bacterial supernatant:

[0028] Take Lactobacillus rhamnosus CGMCC 1.577 strain, inoculate to MRS solid medium, 37℃ incubator inverted culture for 24 hours, pick up the better morphology of single colony inoculated into MRS liquid medium for activation, 37℃, 150r / min culture 24 hours, then centrifugation, 5000xg, 10min centrifugation, the supernatant was filtered by 0.22μm bacterial filter to obtain Lactobacillus rhamnosus supernatant, 4℃ sealed for standby.

[0029] Take Lactobacillus rhamnosus CGMCC 1.577 strain, inoculate to MRS solid medium, 37℃ incubator inverted culture for 24 hours, pick up the better morphology of single colony inoculated into MRS liquid medium for activation, 37℃, 150r / min culture 24 hours, then centrifugation, 5000xg, 10min centrifugation, the supernatant was filtered by 0.22μm bacterial filter to obtain Lactobacillus rhamnosus supernatant, 4℃ sealed for standby.

[0030] Preparation of anti-infection gel:

[0031] The water is heated to 30℃, the water soluble vitamin E nicotinate, vitamin E is added while stirring, the temperature is raised to 40℃ after stirring for 30min, then Lactobacillus rhamnosus supernatant, Lactobacillus acidophilus supernatant, Sophora flavescens essential oil, hyaluronic acid, sodium carboxymethyl cellulose are added and stirred for 2 hours, and then the intrauterine anti-infection gel is obtained.

[0032] Example 2

[0033] Only containing Lactobacillus rhamnosus CGMCC 1.577, the others are the same as example 1.

[0034] The composition of an intrauterine anti-infection anti-adhesion gel:

[0035] The raw materials of the gel are, by mass percentage, Lactobacillus rhamnosus supernatant 6%, Sophora flavescens essential oil 25%, hyaluronic acid 25%, water soluble vitamin E nicotinate 4%, vitamin E 2.5%, sodium carboxymethyl cellulose 2.5%, water 35%;

[0036] Preparation of the bacterial supernatant:

[0037] Take Lactobacillus rhamnosus CGMCC 1.577 strain, inoculate to MRS solid medium, 37℃ incubator inverted culture for 24 hours, pick up the better morphology of single colony inoculated into MRS liquid medium for activation, 37℃, 150r / min culture 24 hours, then centrifugation, 5000xg, 10min centrifugation, the supernatant was filtered by 0.22μm bacterial filter to obtain Lactobacillus rhamnosus supernatant, 4℃ sealed for standby.

[0038] Preparation of the anti-infection gel:

[0039] Heat water to 30℃, add water-soluble vitamin E nicotinate and vitamin E while stirring, and then increase the temperature to 40℃ after stirring for 30 min. Then add Lactobacillus rhamnosus supernatant, Sophora flavescens essential oil, hyaluronic acid, and sodium carboxymethyl cellulose, and stir for 2 hours to obtain the anti-infection gel for intrauterine use.

[0040] Example 3

[0041] Only Lactococcus lactis CGMCC 1.15072 is contained, and the other components are the same as in Example 1.

[0042] Composition of an anti-infection anti-adhesion gel for intrauterine use:

[0043] The raw materials of the gel are, by mass percentage, Lactococcus lactis supernatant 6%, Sophora flavescens essential oil 25%, hyaluronic acid 25%, water-soluble vitamin E nicotinate 4%, vitamin E 2.5%, sodium carboxymethyl cellulose 2.5%, and water 35%.

[0044] Preparation of the bacterial supernatant:

[0045] Thaw Lactococcus lactis CGMCC 1.15072 strain, inoculate it on MRS solid medium, and culture it in an inverted incubator at 30℃ for 24 hours. Pick single colonies with good morphology, inoculate them into MRS liquid medium for activation, and culture them at 37℃ and 150 r / min for 24 hours. Then centrifuge, centrifuge at 5000 x g for 10 min, filter the supernatant with a 0.22 μm bacterial filter to obtain Lactococcus lactis supernatant, and store it at 4℃ in a sealed container for standby use.

[0046] Preparation of the anti-infection gel:

[0047] Heat water to 30℃, add water-soluble vitamin E nicotinate and vitamin E while stirring, and then increase the temperature to 40℃ after stirring for 30 min. Then add Lactococcus lactis supernatant, Sophora flavescens essential oil, hyaluronic acid, and sodium carboxymethyl cellulose, and stir for 2 hours to obtain the anti-infection gel for intrauterine use.

[0048] Example 4

[0049] Composition of an anti-infection anti-adhesion gel for intrauterine use:

[0050] The raw materials of the gel are, by mass percentage, Lactococcus lactis supernatant 6%, Sophora flavescens essential oil 25%, hyaluronic acid 25%, water-soluble vitamin E nicotinate 4%, vitamin E 2.5%, sodium carboxymethyl cellulose 2.5%, and water 35%.

[0051] Preparation of the bacterial supernatant:

[0052] Lactobacillus rhamnosus CGMCC 1.484 strain was thawed, inoculated on MRS solid medium, and cultured in an inverted incubator at 37°C for 24 hours. Single colonies with good morphology were picked and inoculated into MRS liquid medium for activation, and cultured at 37°C and 150 r / min for 24 hours. Then, centrifugation was performed at 5000 x g for 10 minutes, and the supernatant was filtered with a 0.22 μm bacterial filter to obtain Lactobacillus rhamnosus supernatant, which was stored at 4°C for later use.

[0053] Lactobacillus rhamnosus CGMCC 1.484 strain was thawed, inoculated on MRS solid medium, and cultured in an inverted incubator at 37°C for 24 hours. Single colonies with good morphology were picked and inoculated into MRS liquid medium for activation, and cultured at 37°C and 150 r / min for 24 hours. Then, centrifugation was performed at 5000 x g for 10 minutes, and the supernatant was filtered with a 0.22 μm bacterial filter to obtain Lactobacillus rhamnosus supernatant, which was stored at 4°C for later use.

[0054] Preparation of anti-infection gel:

[0055] Water was heated to 30°C, and water-soluble vitamin E nicotinate and vitamin E were added while stirring. After stirring for 30 minutes, the temperature was raised to 40°C, and Lactobacillus rhamnosus supernatant, Lactobacillus lactis supernatant, Sophora flavescens essential oil, hyaluronic acid, and sodium carboxymethyl cellulose were added and stirred for 2 hours to obtain an anti-infection gel for intrauterine use.

[0056] Example 5

[0057] Composition of an anti-infection anti-adhesion gel for intrauterine use:

[0058] The raw materials of the gel are, by mass percentage, Lactobacillus rhamnosus supernatant 3%, Lactobacillus lactis supernatant 3%, Sophora flavescens essential oil 25%, hyaluronic acid 25%, water-soluble vitamin E nicotinate 4%, vitamin E 2.5%, sodium carboxymethyl cellulose 2.5%, and water 35%.

[0059] Preparation of the bacterial supernatant:

[0060] Lactobacillus rhamnosus CGMCC 1.484 strain was thawed, inoculated on MRS solid medium, and cultured in an inverted incubator at 37°C for 24 hours. Single colonies with good morphology were picked and inoculated into MRS liquid medium for activation, and cultured at 37°C and 150 r / min for 24 hours. Then, centrifugation was performed at 5000 x g for 10 minutes, and the supernatant was filtered with a 0.22 μm bacterial filter to obtain Lactobacillus rhamnosus supernatant, which was stored at 4°C for later use.

[0061] Lactococcus lactis strain CGMCC 1.62 was thawed, inoculated on MRS solid medium, and cultured in an inverted incubator at 30℃ for 24 hours. Single colonies with good morphology were selected and inoculated into MRS liquid medium for activation, and cultured at 37℃ and 150 r / min for 24 hours. Then, the supernatant was obtained by centrifugation at 5,000 x g for 10 minutes, and filtered through a 0.22 μm bacterial filter to obtain Lactococcus lactis supernatant, which was stored at 4℃ for later use.

[0062] Preparation of an anti-infection gel:

[0063] Water was heated to 30℃, and water-soluble vitamin E nicotinate and vitamin E were added while stirring. After stirring for 30 minutes, the temperature was raised to 40℃, and plant extract, Sophora flavescens essential oil, hyaluronic acid, and sodium carboxymethyl cellulose were added and stirred for 2 hours to obtain an anti-infection gel for intrauterine use.

[0064] Example 6

[0065] Composition of an anti-infection anti-adhesion gel for intrauterine use:

[0066] The raw materials of the gel are, by mass percentage, Sophora flavescens essential oil 25%, hyaluronic acid 25%, water-soluble vitamin E nicotinate 4%, vitamin E 2.5%, sodium carboxymethyl cellulose 2.5%, and water 41%.

[0067] Preparation of an anti-infection gel:

[0068] Water was heated to 30℃, and water-soluble vitamin E nicotinate and vitamin E were added while stirring. After stirring for 30 minutes, the temperature was raised to 40℃, and Sophora flavescens essential oil, hyaluronic acid, and sodium carboxymethyl cellulose were added and stirred for 2 hours to obtain an anti-infection gel for intrauterine use.

[0069] Example 7

[0070] Preparation of a mouse model of endometritis, 45 female BALB / c mice aged 8-10 weeks were selected. They were raised under conditions of 22-24℃, with humidity maintained at 50%, and were provided with sufficient clean drinking water and suitable feed.

[0071] Antibiotic sensitivity test was performed on the E. coli strain, and E. coli sensitive to ampicillin was selected and added to 50 mL of LB liquid medium in a constant temperature shaker at 37℃ and 120 rpm overnight. Then, 0.5 mL of the bacterial solution was added to 50 mL of LB liquid medium, and cultured at 37℃ and 120 rpm for 5 hours. The bacterial solution was centrifuged at 4,000 x g for 10 minutes, and the E. coli was resuspended in sterile PBS to a concentration of 1 x 10 9 CFU / 100 μL PBS for later use.

[0072] 45 mice were randomly divided into 9 groups, 5 mice in each group, respectively blank group, E. coli group, experimental group (Example 1-6 group), ampicillin group; on the 1st, 3rd, 5th day of the experiment, the blank group was given normal saline 100 μL intrauterine, the E. coli group, the experimental group and the ampicillin group were injected with 1×10 9 CFU / mL E. coli 100 μL to construct an endometritis mouse model; on the 7th-14th day of the experiment, the blank group and the E. coli group were given normal saline 100 μL / day intrauterine, the experimental group was given anti-infection gel prepared by Example 1-6 group 100 μL / day intrauterine, the ampicillin group mice were given 100 μL ampicillin (1 mg / mL); on the 15th day of the experiment, the mice were euthanized, and the uterus was collected for subsequent testing.

[0073] Example 8

[0074] The uterus tissue obtained in Example 7 was collected and stored in 4% paraformaldehyde for 48 hours, paraffin-embedded, sectioned, deparaffinized and HE stained, the inflammatory cell infiltration was evaluated, the number of endometrial plasma cells and lymphocytes in 5 high power fields (HPF) was evaluated and averaged, the changes of interstitial cells and epithelial cells were evaluated, whether the interstitial cells became fusiform, the interstitial congestion and edema, whether the endometrial epithelium existed in multiple layers and the cell nucleus enlarged, according to the degree of diffusion, it was divided into -, +, ++ and +++. And CD138 immunohistochemical detection was carried out at the same time, the number of cells with brownish yellow cell membrane / cytoplasm in each high power field (HPF) was counted, and more than 5 / HPF was considered positive (+). The results are shown in Table 1.

[0075] Table 1 Pathological detection of endometritis

[0076] Group Inflammatory cells (number / HPF) Interstitial and epithelial reactive changes CD138 Blank group 2.10±1.24 - - E. coli group 52.10±16.08 +++ + Example 1 group 6.42 ± 2.85 1，2 ]] - - Example 2 group 19.50 ± 3.80 1，2，3 ]] + + Example 3 group 45.22 ± 16.84 1，2，3 ]] ++ + Example 4 group 44.77 ± 19.75 1，2，3 ]] ++ + Example 5 group 18.18 ± 6.57 1，2，3 ]] + + Example 6 group 50.10 ± 20.44 1，3 ]] +++ + Ampicillin group 5.10±3.32 - -

[0077] Note: t test, 1: P<0.05 (compared with the blank group); 2: P<0.05 (compared with the E. coli group); 3: P<0.05 (compared with the ampicillin group)

[0078] Example 9

[0079] The uterus obtained in Example 7 was washed with 1 ml of sterile normal saline using a sterile syringe. After dilution, 100 μL of the diluent was spread on LB solid plate, 37°C, aerobic culture for 24 hours. The colony forming units were counted. The results are shown in Table 2.

[0080] Table 2 Comparison of E. coli load in mouse uterus

[0081] Group log10 CFU / ml Blank group 1.22±0.15 E. coli group 4.86±0.42 Example 1 group 2.37 ± 0.47 1，2 ]] Example 2 group 3.76 ± 0.71 1，2，3 ]] Example 3 group 4.37 ± 0.40 1，2，3 ]] Example 4 group 4.25 ± 0.64 1，2，3 ]] Example 5 group 3.55 ± 0.39 1，2，3 ]] Example 6 group 4.77 ± 0.71 1，3 ]] Ampicillin group 2.31±0.53

[0082] Note: t test, 1: P < 0.05 (compared with the blank group); 2: P < 0.05 (compared with the E. coli group); 3: P < 0.05 (compared with the ampicillin group)

[0083] Example 10

[0084] The collected uterine tissue was taken 1 cm 3 Add 10 ml of physiological saline to grind, and wash, block, sample, capture antibody, color development and detect TNF-α and IL-1β according to the requirements of the ELISA kit. The results are shown in Table 3.

[0085] Table 3 Immune factor determination results

[0086] Group TNF-α (ng / ml) IL-1β (ng / ml) Blank group 1.10±0.18 1.61±0.21 E. coli group 2.18±0.34 2.65±0.57 Example 1 group 1.26 ± 0.40 1，2 ]] 1.69 ± 0.29 1，2 ]] Example 2 group 1.70 ± 0.38 1，2，3 ]] 2.00 ± 0.18 1，2，3 ]] Example 3 group 1.97 ± 0.35 1，2，3 ]] 2.45 ± 0.45 1，2，3 ]] Example 4 group 1.89 ± 0.37 1，2，3 ]] 2.34 ± 0.27 1，2，3 ]] Example 5 group 1.68 ± 0.30 1，2，3 ]] 1.97 ± 0.29 1，2，3 ]] Example 6 group 2.09 ± 0.21 1，3 ]] 2.49 ± 0.33 1，3 ]] Ampicillin group 1.22±0.23 1.71±0.38

[0087] Note: t test, 1: P < 0.05 (compared with the blank group); 2: P < 0.05 (compared with the E. coli group); 3: P < 0.05 (compared with the ampicillin group)

[0088] Example 10

[0089] When the mouse uterus was dissected, it was also found that the uterus of the blank group mouse had no adhesion, and the degree of uterine cavity adhesion of the ampicillin group and the example 1 group was obviously lighter than that of the example 2-6 group, and no obvious adhesion band component was found. The membrane-like adhesion component of the E. coli group was the most.

[0090] In summary, the results show that the intrauterine inflammatory condition of the mouse in the example group is significantly higher than that in the blank group, among which the example 2 group and the example 5 group (containing lactobacillus rhamnosus CGMCC 1.577) can significantly improve the intrauterine inflammatory condition, while the example 3 group (containing lactobacillus lactis CGMCC 1.15072) and the example 5 group (containing other types of lactobacillus rhamnosus and lactobacillus lactis CGMCC 1.15072) have no obvious effect, which shows that the fermentation product of lactobacillus rhamnosus CGMCC 1.577 has its unique property, can produce synergistic effect with lactobacillus lactis CGMCC 1.15072, and thus better strengthens the process of gel resisting endometritis, thereby improving the treatment effect.

[0091] In summary, in the research and practice process, the research team of the present application found that the fermentation product of lactobacillus rhamnosus CGMCC 1.577 has its unique property, can produce synergistic effect with lactobacillus lactis CGMCC 1.15072, and can have excellent reduction of intrauterine inflammatory cell infiltration, interstitial and epithelial reactivity change, reduction of the number of miscellaneous bacteria, reduction of inflammatory factors, and better improvement of endometritis.

[0092] The description of an embodiment or examples using terms such as "one embodiment", "some embodiments", "an example", "a specific example", or "some examples" means that a particular feature, structure, material, or characteristic is included in at least one embodiment or example of the present application. The illustrative appearances of the above-mentioned terms in various places in the specification are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics can be combined in any suitable manner in one or more embodiments or examples.

[0093] While embodiments of the application have been shown and described, it is to be understood that the application is not limited to the details of the embodiments described, since the scope of the application will be defined with regard to the appended claims, along with their equivalents.

Claims

1. An anti-infective anti-adhesion gel for intrauterine use, characterized in that it comprises: The raw materials of the vaginal gel are, by mass percentage, Lactobacillus rhamnosus supernatant 3%, Lactococcus lactis supernatant 3%, Sophora flavescens essential oil 25%, hyaluronic acid 25%, water-soluble vitamin E nicotinic acid ester 4%, vitamin E 2.5%, sodium carboxymethyl cellulose 2.5%, and water 35%; Preparation of the Lactobacillus rhamnosus supernatant: the Lactobacillus rhamnosus CGMCC 1.577 strain was thawed, inoculated on MRS solid medium, and cultured in a 37℃ incubator for 24 hours, single colonies with good morphology were picked and inoculated in MRS liquid medium for activation, and cultured at 37℃ and 150 r / min for 24 hours, then centrifuged at 5000 x g for 10 minutes, and the supernatant was filtered with a 0.22 μm bacterial filter to obtain the Lactobacillus rhamnosus supernatant, which was stored at 4℃ for later use; Preparation of the Lactococcus lactis supernatant: the Lactococcus lactis CGMCC 1.15072 strain was thawed, inoculated on MRS solid medium, and cultured in a 30℃ incubator for 24 hours, single colonies with good morphology were picked and inoculated in MRS liquid medium for activation, and cultured at 30℃ and 200 r / min for 24 hours, then centrifuged at 5000 x g for 10 minutes, and the supernatant was filtered with a 0.22 μm bacterial filter to obtain the Lactococcus lactis supernatant, which was stored at 4℃ for later use.

2. An anti-adhesion infection preventing gel for intrauterine use according to claim 1, characterized in that, The preparation steps include at least the following: heating water to 30℃, adding water-soluble vitamin E nicotinic acid ester and vitamin E while stirring, stirring for 30 minutes, then increasing the temperature to 40℃, and adding Lactobacillus rhamnosus supernatant, Lactococcus lactis supernatant, Sophora flavescens essential oil, hyaluronic acid, and sodium carboxymethyl cellulose, and stirring for 2 hours to obtain the anti-infection and anti-adhesion gel for intrauterine use.