An anti-infective anti-adhesion gel for intrauterine use and a method for its preparation

Abstract

The application discloses an anti-infection anti-adhesion gel for intrauterine use and a preparation method thereof, which comprises lactobacillus rhamnosus CGMCC 1.577 and lactococcus lactis CGMCC 1.15072. The fermentation product of the lactobacillus rhamnosus CGMCC 1.577 has a unique property, can produce a synergistic effect with the lactococcus lactis CGMCC 1.15072, can have a relatively excellent effect of reducing cell infiltration, interstitial and epithelial reactivity changes, reducing the number of miscellaneous bacteria and reducing inflammatory factors, and can better improve endometritis symptoms.

Description

Technical Field

[0001] This invention relates to daily chemical products for women's hygiene and health, and more particularly to an intrauterine anti-infection and anti-adhesion gel and its preparation method. Background Technology

[0002] Endometritis, a common gynecological disease, affects the health of a large number of women worldwide. Statistics show that the incidence of endometritis among sexually active women of reproductive age is increasing year by year, seriously impacting their reproductive health and quality of life. If left untreated, endometritis can lead to serious complications such as infertility and ectopic pregnancy, and may also cause problems such as intrauterine adhesions, chronic pelvic pain, and pyometra, further aggravating the patient's suffering and imposing a heavy economic burden on families and society.

[0003] Currently, clinical treatments for endometritis mainly include systemic and local therapies. Systemic therapies often involve intravenous infusion, intramuscular injection, or oral administration of antibiotics. However, due to the unique physiological structure of the uterus, these drugs often fail to effectively reach the affected area, resulting in unsatisfactory efficacy. Local therapies typically involve flushing the uterus with antibiotic or disinfectant solutions. While these methods can kill pathogens and remove inflammatory secretions to some extent, the widespread use of antibiotics has led to an increase in drug-resistant strains, gradually reducing the effectiveness of treatment. Furthermore, the bactericidal effect of the solution after flushing is relatively short-lived, requiring frequent flushing to achieve optimal therapeutic results. This not only increases the workload of medical staff but also causes considerable inconvenience for patients.

[0004] Among the many drug formulations for treating endometritis, gels have received widespread attention due to their unique advantages. Gels possess good adhesion, allowing them to remain in the uterus for a longer period, prolonging the contact time between the drug and the lesion site, thereby improving the drug's efficacy. However, the effects of existing gels for treating endometritis are not ideal.

[0005] To improve treatment outcomes, the development of a highly efficient, stable, and antibacterial intrauterine gel for treating endometritis and preventing adhesions is of significant clinical importance and practical necessity.

[0006] This invention aims to provide an intrauterine anti-infective and anti-adhesion gel and its preparation method, which is expected to bring new treatment options to a large number of patients with endometritis and those who may have endometritis after surgery, make up for the shortcomings of existing treatment methods, and has significant clinical significance and market value. Summary of the Invention

[0007] The purpose of this invention is to provide a method for preparing an intrauterine anti-infective and anti-adhesion gel. The resulting intrauterine anti-infective gel can effectively improve the intrauterine inflammatory state, reduce the number of intrauterine bacteria, increase endometrial thickness, and alleviate intrauterine adhesions, thereby treating endometritis.

[0008] All bacteria used in this patent are commercially available.

[0009] The Lactobacillus rhamnosus strain is deposited at the China General Microbiological Culture Collection Center with accession number CGMCC 1.577.

[0010] The Lactobacillus rhamnosus strain is deposited at the China General Microbiological Culture Collection Center with accession number CGMCC 1.484.

[0011] The Lactococcus lactis strain is deposited at the China General Microbiological Culture Collection Center with accession number CGMCC 1.15072.

[0012] The Lactococcus lactis strain is deposited at the China General Microbiological Culture Collection Center with accession number CGMCC 1.62.

[0013] During their research and practice, the applicant's research team discovered that the fermentation product of *Lactobacillus rhamnosus* CGMCC1.577 has unique properties and can synergistically interact with *Lactococcus lactis* CGMCC1.15072. This synergistic effect significantly reduces intrauterine inflammatory cell infiltration, alters in stromal and epithelial reactivity, decreases the number of contaminating bacteria, and reduces inflammatory factors, thus effectively improving endometritis symptoms. The technical problem to be solved by this invention can be achieved through the following technical solution.

[0014] Composition of an intrauterine anti-infective gel:

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

[0016] Preparation of the bacterial supernatant:

[0017] Thaw Lactobacillus rhamnosus CGMCC 1.577 strain, inoculate onto MRS solid medium, and incubate upside down at 37℃ for 24 hours. Select single colonies with good morphology and inoculate them into MRS liquid medium for activation. Incubate at 37℃ and 150 r / min for 24 hours, then centrifuge at 5000×g for 10 min. Filter the supernatant through a 0.22 μm bacterial filter to obtain Lactobacillus rhamnosus supernatant, and store it in a sealed container at 4℃ for later use.

[0018] Thaw the Lactococcus lactis CGMCC 1.15072 strain, inoculate it onto MRS solid medium, and incubate it upside down in a 30℃ incubator for 24 hours. Select single colonies with good morphology and inoculate them into MRS liquid medium for activation. Incubate at 30℃ and 200 r / min for 24 hours, then centrifuge at 5000×g for 10 min. Filter the supernatant through a 0.22μm bacterial filter to obtain the Lactococcus lactis supernatant, and store it in a sealed container at 4℃ for later use.

[0019] Preparation of anti-infective gel:

[0020] Heat water to 30°C, add water-soluble vitamin E nicotinic acid ester and vitamin E while stirring, stir for 30 minutes, then raise the temperature to 40°C, add Lactobacillus rhamnosus supernatant, Lactococcus lactis supernatant, Sophora flavescens essential oil, hyaluronic acid and sodium carboxymethyl cellulose and stir for 2 hours to obtain intrauterine anti-infection gel.

[0021] Advantages of this invention:

[0022] (1) The fermentation product of Lactobacillus rhamnosus CGMCC 1.577 has its unique characteristics. It can have a synergistic effect with Lactococcus lactis CGMCC 1.15072. It can effectively reduce the infiltration of inflammatory cells in the endometrium, the changes in the reactivity of the stroma and epithelium, reduce the number of miscellaneous bacteria, reduce inflammatory factors, and can effectively improve the symptoms of endometritis. Detailed Implementation

[0023] The embodiments of the present invention are described in detail below. These embodiments are only used to explain the present invention and should not be construed as limiting the present invention.

[0024] Example 1

[0025] Composition of an intrauterine anti-infective and anti-adhesion gel:

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

[0027] Preparation of the bacterial supernatant:

[0028] Thaw Lactobacillus rhamnosus CGMCC 1.577 strain, inoculate onto MRS solid medium, and incubate upside down at 37℃ for 24 hours. Select single colonies with good morphology and inoculate them into MRS liquid medium for activation. Incubate at 37℃ and 150 r / min for 24 hours, then centrifuge at 5000×g for 10 min. Filter the supernatant through a 0.22 μm bacterial filter to obtain Lactobacillus rhamnosus supernatant, and store it in a sealed container at 4℃ for later use.

[0029] Thaw the Lactococcus lactis CGMCC 1.15072 strain, inoculate it onto MRS solid medium, and incubate it upside down in a 30℃ incubator for 24 hours. Select single colonies with good morphology and inoculate them into MRS liquid medium for activation. Incubate at 30℃ and 200 r / min for 24 hours, then centrifuge at 5000×g for 10 min. Filter the supernatant through a 0.22μm bacterial filter to obtain the Lactococcus lactis supernatant, and store it in a sealed container at 4℃ for later use.

[0030] Preparation of anti-infective gel:

[0031] Heat water to 30°C, add water-soluble vitamin E nicotinic acid ester and vitamin E while stirring, stir for 30 minutes, then raise the temperature to 40°C, add Lactobacillus rhamnosus supernatant, Lactococcus lactis supernatant, Sophora flavescens essential oil, hyaluronic acid and sodium carboxymethyl cellulose and stir for 2 hours to obtain intrauterine anti-infection gel.

[0032] Example 2

[0033] It contains only Lactobacillus rhamnosus CGMCC 1.577, and everything else is the same as in Example 1.

[0034] Composition of an intrauterine anti-infective and anti-adhesion gel:

[0035] The raw materials for the gel are, by mass percentage: 6% Lactobacillus rhamnosus supernatant, 25% Sophora flavescens essential oil, 25% hyaluronic acid, 4% water-soluble vitamin E nicotinic acid ester, 2.5% vitamin E, 2.5% sodium carboxymethyl cellulose, and 35% water.

[0036] Preparation of the bacterial supernatant:

[0037] Thaw Lactobacillus rhamnosus CGMCC 1.577 strain, inoculate onto MRS solid medium, and incubate upside down at 37℃ for 24 hours. Select single colonies with good morphology and inoculate them into MRS liquid medium for activation. Incubate at 37℃ and 150 r / min for 24 hours, then centrifuge at 5000×g for 10 min. Filter the supernatant through a 0.22 μm bacterial filter to obtain Lactobacillus rhamnosus supernatant, and store it in a sealed container at 4℃ for later use.

[0038] Preparation of anti-infective gel:

[0039] Heat water to 30°C, add water-soluble vitamin E nicotinic acid ester and vitamin E while stirring, stir for 30 minutes, then raise the temperature to 40°C, add Lactobacillus rhamnosus supernatant, Sophora flavescens essential oil, hyaluronic acid and sodium carboxymethyl cellulose and stir for 2 hours to obtain intrauterine anti-infection gel.

[0040] Example 3

[0041] It contains only Lactococcus lactis CGMCC 1.15072, and everything else is the same as in Example 1.

[0042] Composition of an intrauterine anti-infective and anti-adhesion gel:

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

[0044] Preparation of the bacterial supernatant:

[0045] Thaw the Lactococcus lactis CGMCC 1.15072 strain, inoculate it onto MRS solid medium, and incubate it upside down in a 30℃ incubator for 24 hours. Select single colonies with good morphology and inoculate them into MRS liquid medium for activation. Incubate at 37℃ and 150 r / min for 24 hours, then centrifuge at 5000×g for 10 min. Filter the supernatant through a 0.22μm bacterial filter to obtain the Lactococcus lactis supernatant, and store it in a sealed container at 4℃ for later use.

[0046] Preparation of anti-infective gel:

[0047] Heat water to 30°C, add water-soluble vitamin E nicotinic acid ester and vitamin E while stirring, stir for 30 minutes, then raise the temperature to 40°C, add lactococcus lactis supernatant, sophora flavescens essential oil, hyaluronic acid and sodium carboxymethyl cellulose and stir for 2 hours to obtain intrauterine anti-infection gel.

[0048] Example 4

[0049] Composition of an intrauterine anti-infective and anti-adhesion gel:

[0050] The raw materials for the gel are, by mass percentage: 3% Lactobacillus rhamnosus supernatant, 3% Lactococcus lactis supernatant, 25% Sophora flavescens essential oil, 25% hyaluronic acid, 4% water-soluble vitamin E nicotinic acid ester, 2.5% vitamin E, 2.5% sodium carboxymethyl cellulose, and 35% water.

[0051] Preparation of the bacterial supernatant:

[0052] Thaw Lactobacillus rhamnosus CGMCC 1.484 strain, inoculate it onto MRS solid medium, and incubate it upside down in an incubator at 37°C for 24 hours. Select single colonies with good morphology and inoculate them into MRS liquid medium for activation. Incubate at 37°C and 150 r / min for 24 hours, then centrifuge at 5000×g for 10 min. Filter the supernatant through a 0.22 μm bacterial filter to obtain Lactobacillus rhamnosus supernatant, and store it in a sealed container at 4°C for later use.

[0053] Thaw the Lactococcus lactis CGMCC 1.15072 strain, inoculate it onto MRS solid medium, and incubate it upside down in a 30℃ incubator for 24 hours. Select single colonies with good morphology and inoculate them into MRS liquid medium for activation. Incubate at 37℃ and 150 r / min for 24 hours, then centrifuge at 5000×g for 10 min. Filter the supernatant through a 0.22μm bacterial filter to obtain the Lactococcus lactis supernatant, and store it in a sealed container at 4℃ for later use.

[0054] Preparation of anti-infective gel:

[0055] Heat water to 30°C, add water-soluble vitamin E nicotinic acid ester and vitamin E while stirring, stir for 30 minutes, then raise the temperature to 40°C, add Lactobacillus rhamnosus supernatant, Lactococcus lactis supernatant, Sophora flavescens essential oil, hyaluronic acid and sodium carboxymethyl cellulose and stir for 2 hours to obtain intrauterine anti-infection gel.

[0056] Example 5

[0057] Composition of an intrauterine anti-infective and anti-adhesion gel:

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

[0059] Preparation of the bacterial supernatant:

[0060] Thaw Lactobacillus rhamnosus CGMCC 1.577 strain, inoculate onto MRS solid medium, and incubate upside down at 37℃ for 24 hours. Select single colonies with good morphology and inoculate them into MRS liquid medium for activation. Incubate at 37℃ and 150 r / min for 24 hours, then centrifuge at 5000×g for 10 min. Filter the supernatant through a 0.22 μm bacterial filter to obtain Lactobacillus rhamnosus supernatant, and store it in a sealed container at 4℃ for later use.

[0061] Thaw the Lactococcus lactis CGMCC 1.62 strain, inoculate it onto MRS solid medium, and incubate it upside down in a 30℃ incubator for 24 hours. Select single colonies with good morphology and inoculate them into MRS liquid medium for activation. Incubate at 37℃ and 150 r / min for 24 hours, then centrifuge at 5000×g for 10 min. Filter the supernatant through a 0.22μm bacterial filter to obtain the Lactococcus lactis supernatant, and store it in a sealed container at 4℃ for later use.

[0062] Preparation of anti-infective gel:

[0063] Heat water to 30°C, add water-soluble vitamin E nicotinic acid ester and vitamin E while stirring, stir for 30 minutes, then raise the temperature to 40°C, add plant extracts, sophora flavescens essential oil, hyaluronic acid and sodium carboxymethyl cellulose and stir for 2 hours to obtain intrauterine anti-infection gel.

[0064] Example 6

[0065] Composition of an intrauterine anti-infective and anti-adhesion gel:

[0066] The raw materials for the gel are, by weight percentage: 25% Sophora flavescens essential oil, 25% hyaluronic acid, 4% water-soluble vitamin E nicotinic acid ester, 2.5% vitamin E, 2.5% sodium carboxymethyl cellulose, and 41% water;

[0067] Preparation of anti-infective gel:

[0068] Heat water to 30°C, add water-soluble vitamin E nicotinic acid ester and vitamin E while stirring, stir for 30 minutes, then raise the temperature to 40°C, add sophora flavescens essential oil, hyaluronic acid and sodium carboxymethyl cellulose and stir for 2 hours to obtain intrauterine anti-infection gel.

[0069] Example 7

[0070] To establish a mouse model of endometritis, 45 female BALB / c mice aged 8-10 weeks were selected. They were housed at 22-24℃ with 50% humidity, and provided with ample clean drinking water and suitable feed.

[0071] Antibiotic susceptibility testing was performed on *E. coli* strains. *E. coli* strains sensitive to ampicillin were added to LB broth and incubated overnight at 37°C and 120 rpm on a shaker. Then, 0.5 mL of the bacterial suspension was added to 50 mL of LB broth and incubated for 5 hours at 37°C and 120 rpm on a shaker. The suspension was then centrifuged at 4000×g for 10 min, and the *E. coli* were resuspended in sterile PBS to achieve a concentration of 1×10⁻⁶. 9 Prepare CFU / 100μL PBS for later use.

[0072] Forty-five mice were randomly divided into nine groups of five mice each: a control group, an E. coli group, an experimental group (groups 1-6 in Examples), and an ampicillin group. On days 1, 3, and 5 of the experiment, the control group received 100 μL of physiological saline intrauterinely, while the E. coli group, experimental group, and ampicillin group received 1 × 10⁻⁶ mice intrauterinely. 9 A mouse model of endometritis was established using 100 μL of CFU / mL Escherichia coli. From day 7 to day 14 of the experiment, mice in the control group and the E. coli group were given 100 μL / day of physiological saline intrauterinely, while mice in the experimental group were given 100 μL / day of the anti-infection gel prepared in Examples 1-6, and mice in the ampicillin group were given 100 μL of ampicillin (1 mg / mL). On day 15 of the experiment, the mice were euthanized and the uterus was collected for subsequent testing.

[0073] Example 8

[0074] Uterine tissue obtained in Example 7 was preserved in 4% paraformaldehyde for 48 hours, embedded in paraffin, dewaxed, and stained with hematoxylin and eosin (HE) to assess inflammatory cell infiltration. The number of plasma cells and lymphocytes per 5 high-power fields (HPF) was assessed and averaged. Changes in stromal cells and epithelial cells were also assessed, including whether stromal cells became spindle-shaped, the presence of stromal congestion and edema, and the presence of stratified endometrial epithelium and enlarged nuclei. Based on the degree of diffusion, the results were categorized as -, +, ++, and +++. Simultaneously, CD138 immunohistochemical detection was performed, counting the number of cells showing a brownish-yellow color in the cell membrane / cytoplasm per HPF; a count greater than 5 cells per HPF was considered positive (+). The results are shown in Table 1.

[0075] Table 1. Pathological examination of endometritis

[0076] Blank group 2.10±1.24 - - Escherichia coli group 52.10±16.08 +++ + Example 1 Group <![CDATA[6.42±2.85 1，2 ]]> - - Example 2 group <![CDATA[19.50±3.80 1，2，3 ]]> + + Example 3 Group <![CDATA[45.22±16.84 1，2，3 ]]> ++ + Example 4 group <![CDATA[44.77±19.75 1，2，3 ]]> ++ + Example 5 group <![CDATA[18.18±6.57 1，2，3 ]]> + + Example 6 group <![CDATA[50.10±20.44 1，3 ]]> +++ + Ampicillin group 5.10±3.32 - -

[0077] Note: t-test, 1: P < 0.05 (compared with the control 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 irrigated with 1 ml of sterile physiological saline using a sterile syringe. After serial dilution, 100 μL of the diluted solution was spread onto LB agar plates and incubated aerobically at 37°C for 24 hours. Colony-forming units were counted. The results are shown in Table 2.

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

[0081] Blank group 1.22±0.15 Escherichia coli group 4.86±0.42 Example 1 Group <![CDATA[2.37±0.47 1，2 ]]> Example 2 group <![CDATA[3.76±0.71 1，2，3 ]]> Example 3 Group <![CDATA[4.37±0.40 1，2，3 ]]> Example 4 group <![CDATA[4.25±0.64 1，2，3 ]]> Example 5 group <![CDATA[3.55±0.39 1，2，3 ]]> Example 6 group <![CDATA[4.77±0.71 1，3 ]]> Ampicillin group 2.31±0.53

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

[0083] Example 10

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

[0085] Table 3 Results of Immune Factor Measurement

[0086] Blank group 1.10±0.18 1.61±0.21 Escherichia coli group 2.18±0.34 2.65±0.57 Example 1 Group <![CDATA[1.26±0.40 1，2 ]]> <![CDATA[1.69±0.29 1，2 ]]> Example 2 group <![CDATA[1.70±0.38 1，2，3 ]]> <![CDATA[2.00±0.18 1，2，3 ]]> Example 3 Group <![CDATA[1.97±0.35 1，2，3 ]]> <![CDATA[2.45±0.45 1，2，3 ]]> Example 4 group <![CDATA[1.89±0.37 1，2，3 ]]> <![CDATA[2.34±0.27 1，2，3 ]]> Example 5 group <![CDATA[1.68±0.30 1，2，3 ]]> <![CDATA[1.97±0.29 1，2，3 ]]> Example 6 group <![CDATA[2.09±0.21 1，3 ]]> <![CDATA[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 control group); 2: P < 0.05 (compared with the E. coli group); 3: P < 0.05 (compared with the ampicillin group).

[0088] Example 10

[0089] Dissection of the mouse uterus revealed no adhesions in the uterus of the blank group mice. The degree of intrauterine adhesions in the ampicillin group and the Example 1 group was significantly less than that in the Example 2-6 groups, with no obvious adhesion bands observed. The E. coli group had the most membranous adhesions.

[0090] In summary, the results showed that compared with the control group, the uterine inflammation in the mice in the example groups was significantly increased. Specifically, Example 2 and Example 5 (containing *Lactobacillus rhamnosus* CGMCC 1.577) significantly improved the uterine inflammation, while Example 3 (containing *Lactococcus lactis* CGMCC 1.15072) and Example 5 (containing other types of *Lactobacillus rhamnosus* and *Lactococcus lactis* CGMCC 1.15072) showed no significant effect. This indicates that the fermentation product of *Lactobacillus rhamnosus* CGMCC 1.577 has unique properties and can synergistically interact with *Lactococcus lactis* CGMCC 1.15072, thereby enhancing the gel's anti-endometritis process and improving the therapeutic effect.

[0091] In summary, during the research and practice process, the applicant's research team discovered that the fermentation product of Lactobacillus rhamnosus CGMCC 1.577 has unique characteristics and can synergistically interact with Lactococcus lactis CGMCC 1.15072. This synergistic effect can significantly reduce intrauterine inflammatory cell infiltration, altered stromal and epithelial reactivity, decrease the number of miscellaneous bacteria, and reduce inflammatory factors, thus effectively improving endometritis symptoms.

[0092] The terms "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., refer to a specific feature, structure, material, or characteristic described in connection with that embodiment or example, which is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0093] Although embodiments of the invention have been shown and described, those skilled in the art will understand that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents.

Claims

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

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