Urine bacteria transplantation liquid for treating interstitial cystitis as well as preparation method and application of urine bacteria transplantation liquid
Urine microecology of patients with interstitial cystitis was reconstructed through urinary transplantation, which solved the problem of poor effectiveness of existing treatment methods, and achieved effective treatment of interstitial cystitis and significant improvement in quality of life.
Patent Information
- Application Number
- CN202510667574.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-22
- Publication Date
- 2025-08-26
AI Technical Summary
Currently, there is a lack of effective treatment of interstitial cystitis, the existing fecal bacteria transplant treatment is not effective, and the relationship between urine microecology disorder and disease has not been fully explored.
Urine in the urine of healthy people is transplanted. By strictly screening donors and preparing urinary fluid, urinary transplantation is performed to reconstruct the patient's disordered urine microecology, and the urinary bacterial population in healthy urine is used to intervene in the intravesive microenvironment.
It significantly improved the symptoms of frequent urination and pain in patients with interstitial cystitis, restored the diversity and health of urine microecology, reduced chance pathogenic bacteria, improved the quality of life and no adverse reactions such as urinary tract infections were found.
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Figure CN120531771A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of medical technology, and in particular relates to a urine bacteria transplantation solution for treating interstitial cystitis, and a preparation method and application thereof. Background Art
[0002] Interstitial cystitis (IC / BPS) is defined as a chronic, nonbacterial inflammatory bladder disease characterized by frequent urination, urgency, and chronic pelvic pain. The diagnosis of IC is often established through clinical manifestations and physical examination, with ancillary tests such as cystoscopy and urodynamics used to exclude alternative diagnoses. Given the unknown etiology of IC, there are currently no proven treatments. In recent years, there has been extensive research on fecal microbiota transplantation (FMT) for various conditions, including some urinary tract diseases, but the efficacy has been limited. Urinary microbiota transplantation (UMT) involves transplanting specific urinary microbiota from healthy individuals into the patient's bladder to restore the patient's disrupted urinary microbiome. Urine is stored in the bladder for the majority of its duration, from production to excretion. Urine bacteria reside in the bladder for a long time, playing a significant role in bladder homeostasis. Therefore, urinary bacteria transplantation may be an effective intervention for some urinary system diseases that currently have no effective treatment. Currently, there are no reports in domestic and foreign literature on the use of urinary bacteria transplantation to treat urinary system diseases. Summary of the Invention
[0003] The purpose of the present invention is to provide a urine bacteria transplantation solution for treating refractory interstitial cystitis, and a preparation method and application thereof.
[0004] In order to achieve the above objectives, the technical solution adopted by the present invention is:
[0005] From May 2022 to the present, three female patients with refractory interstitial cystitis were admitted to the Central Hospital Affiliated to Jiangnan University. All three patients presented with frequent and urgent urination, two with dysuria, and two with lower abdominal pain. One patient had a history of hypertension, one with anxiety, and two with non-organic sleep disorders; none had other chronic diseases.
[0006] Laboratory tests, instrumental assessments, and specialized examinations were performed on all three patients before and after the urinary bacterial transplant. Laboratory tests included complete blood count, blood biochemistry, urinalysis, and urine culture (to exclude other diseases such as bacterial cystitis). Instrumental assessments included the Self-Rating Depression Scale (SDS), the Self-Rating Anxiety Scale (SAS), the O'Leary-Sant Symptom Index (ICSI), the O'Leary-Sant Problem Index (ICSI), the Urinary Problems Short Form, the Genitourinary Pain Assessment Form, and the Pain Catastrophizing Scale (PCS) (to comprehensively assess patients' quality of life and symptom severity). Specialized examinations included cystoscopy (to exclude bladder cancer and observe the bladder and urethral mucosal condition).
[0007] Urine bacteria collection and analysis:
[0008] (1) Urine collection: 1) Use catheterization to collect urine from the subject. Ask the subject to clean and disinfect the upper 1 / 3 of the inner thigh, perineum, and anus; 2) Wear sterile gloves according to aseptic principles to ensure the operator's hands are clean; 3) Place a sterile drape on the subject's perineum to create a relatively sterile area to prevent contamination; 4) Drain urine through the catheter into a sterile container and temporarily store at 4°C.
[0009] 4. (2) Urine bacterial extraction: 1) Separate the supernatant and urine bacterial pellet by high-speed centrifugation (12,000 rpm, 20 minutes); 2) In a cleanroom, resuspend the urine bacterial pellet in 40 ml of sterile saline to prepare a urine bacterial solution. The urine bacterial pellet includes bacterial precipitate and other components such as urinary tract exfoliated cells. The concentration of the urine bacterial pellet in the urine bacterial transplant solution is 0.01-0.05 g / L.
[0010] (3) Urine bacteria sequencing and analysis methods: DNA was extracted from all urine samples and the V4 region of the 16S rRNA gene of all urinary bacteria was sequenced repeatedly. The relevant primer pair sequences were F: GTGCCAGCMGCCGCGGTAA, R: GGACTACHVGGGTWTCTAAT.
[0011] Urine bacteria transplantation: This study was approved by the medical ethics review of the Central Hospital Affiliated to Jiangnan University (2021-Y-8), and all patients underwent urine bacteria transplantation treatment after signing informed consent.
[0012] (1) Donor screening: Donors should be 18-22 years old, female, and have a healthy diet and lifestyle. All donors need to undergo strict laboratory tests, mainly including: blood routine, coagulation function and C-reactive protein tests; urine routine and urine sediment tests; stool routine and occult blood tests; hepatitis B five items, hepatitis C antibody, syphilis-specific antibody and HIV antibody tests; immune function tests; liver function, blood biochemistry and blood lipid tests; routine urine culture tests; TOUCH four items (rubella, toxoplasmosis, cytomegalovirus, herpes simplex virus) tests; thyroid and parathyroid tests; sex hormone combination tests; tumor marker screening. Donors who pass the laboratory tests need to undergo further urine microecological tests, mainly including: 16SrRNA sequencing analysis of urine bacteria in donor urine samples. Donors with high diversity of urine flora and low abundance of pathogens are preferred. For example, only donors whose abundance of any one of the bacterial species of the genus Lactobacillus, Corynebacterium, Prevotella, Staphylococcus, or Streptococcus is among the top ten in the abundance ranking of all bacterial species are selected.
[0013] (2) Urine transplantation procedure: On the day before the urinary bacterial transplantation, the recipient is to be bladder-washed with 200 mL of 0.9% normal saline. On the day of the urinary bacterial transplantation, the recipient is to be asked to empty the bladder within half an hour before the transplantation operation. The donor urine solution is slowly heated to 37°C in advance. The operator ensures that the prepared donor urine solution (30 mL) is slowly injected into the recipient's bladder under sterile conditions. The infusion speed should not be too fast to avoid irritation or excessive pressure. After the urine solution is fully injected, the recipient is asked to lie supine, left lateral position, right lateral position, prone position and standing position for 10 minutes each to promote urinary bacterial colonization. During the urinary bacterial transplantation operation, the general condition of the recipient is observed at any time, and the body temperature is checked. If there is any abnormality, the operation must be stopped immediately and appropriate treatment must be carried out. Within 24 hours after the first urinary bacterial transplantation operation, the recipient is observed for urinary tract irritation symptoms (such as frequent urination, urgency, pain when urinating, etc.), and the body temperature is checked to rule out infectious fever or allergies and other adverse conditions.
[0014] Beneficial effects of the present invention:
[0015] Healthy individuals possess a unique urinary microbiome, the balance of which is closely linked to health. It is currently believed that the microorganisms found in the urinary microbiome of healthy individuals are primarily slow-growing bacteria, most of which belong to the five major phyla: Firmicutes, Bacteroidetes, Actinobacteria, Fusobacteria, and Proteobacteria. These phyla include Lactobacillus, Corynebacterium, Prevotella, Staphylococcus, and Streptococcus. Numerous studies have suggested that an imbalance in the urinary microbiome may be associated with the onset and progression of disease. One study found a significant decrease in the abundance of Lactobacillus in women with overactive bladder. Furthermore, studies have reported that changes in the abundance of Escherichia coli, Enterococcus, and Staphylococcus are often associated with urinary tract infections.
[0016] Previously, interstitial cystitis was considered a non-inflammatory disease, overlooking the potential link between urinary bacteria and interstitial cystitis. Although studies on the urinary microbiome in patients with interstitial cystitis remain limited, evidence suggests that the diversity and species composition of the urinary microbiome differ significantly between patients with interstitial cystitis and healthy controls. Compared with healthy controls, the urine of patients with interstitial cystitis has a higher abundance of Lactobacillus species and lower microbial diversity and richness. Therefore, we pioneered the use of urinary microbial transplantation to reshape the disturbed urinary microbiome in patients with interstitial cystitis.
[0017] Let’s analyze it based on our experimental results:
[0018] Ding had a history of fecal microbiota transplantation treatment, but there were no obvious changes in his urine microecology before and after the fecal microbiota transplantation, which may be the reason why his symptoms did not improve significantly. However, Ding's urine microecology improved significantly after urine microbiota transplantation, and the distribution of various species was more uniform than after fecal microbiota transplantation. The patient's microbial community structure after urine microbiota transplantation was close to that of the donor. On the other hand, in terms of sampling time, the response of urine microecology after urine microbiota transplantation was faster than that of fecal microbiota transplantation, and the microbial community structure could change faster. After urine microbiota transplantation, the donor microbiota directly contacts the bladder mucosa, changes the microenvironment in the bladder, and thus more effectively corrects the patient's disordered urine microecology. From this point of view, urine microbiota transplantation may be more suitable than fecal microbiota transplantation for intervening in urinary system diseases such as interstitial cystitis. In addition, sequencing results from the three patients after urinary bacterial transplantation indicated a significant decrease in some opportunistic pathogens or harmful bacteria compared to before transplantation. For example, Cupriavidus, a genus that some studies have shown may be positively correlated with systemic inflammatory responses, and other studies have shown that this genus is enriched in prostate cancer samples. Ureaplasma, a common inhabitant of the human reproductive tract, has been increasingly reported to cause severe and even fatal infections in immunocompromised hosts. Anaerobic infections are often caused by multiple microorganisms, most of which include Bacteroides fragilis, with an associated mortality rate exceeding 19%. Pseudomonas, a major pathogen causing urinary tract infections and often resistant to antibiotics, was also found to be an important cause of urinary tract infections and often resistant to antibiotics. After urinary bacterial transplantation, an increase in beneficial commensal bacteria commonly found in healthy people, such as Streptococcus, was also observed. Some scholars have proposed developing Streptococcus as an ideal candidate for oral model probiotics; studies have shown that Dietzia can decompose perfluoroalkyl and polyfluoroalkyl substances, and these synthetic organic halides can enter the human body through contaminated diet, drinking water and environment, and accumulate in the body; Halomonas has been reported as a dominant genus associated with the jejunal and ileal mucosa of goats, and it is speculated that this genus may play a role in promoting intestinal immune development.
[0019] In this study, three patients with interstitial cystitis presented with progressive frequent and painful urination, with no significant improvement after multiple treatments. Urinary microbial transplantation significantly improved their symptoms, and long-term follow-up revealed no significant recurrence, significant relief of anxiety and depression, and no significant abnormalities in related examinations. Furthermore, the patients experienced no urinary tract infections, bacterial translocation, or other serious adverse events previously reported after fecal microbial transplantation throughout the course of treatment. The results of this study strongly suggest that urinary microbial transplantation is safe and effective, and has great potential for treating some urinary tract diseases for which there are currently no effective treatments. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] Figure 1Cystoscopy results before and after urinary bacterial transplantation. (A) Cystoscopy results for Ding before urinary bacterial transplantation. (B) Cystoscopy results for Ding after urinary bacterial transplantation. (C) Cystoscopy results for Lu before urinary bacterial transplantation. (D) Cystoscopy results for Lu after urinary bacterial transplantation. (E) Cystoscopy results for Zhang before urinary bacterial transplantation. (F) Cystoscopy results for Zhang after urinary bacterial transplantation.
[0021] Figure 2 Results of urine microbiome analysis before and after urinary bacterial transplantation in three patients. (AC) Stacked plots of relative abundance of urinary bacteria at the phylum level; (DF) Stacked plots of relative abundance of urinary bacteria at the genus level. All urinary bacterial samples were sequenced twice, and some unidentified species were removed from the results.
[0022] Figure 3 Results of multiple urine microbiome tests from donors Guo and Tang at different times. (AB) Multiple urine microbiome test results from donor Tang, stacked plots of relative abundance of urine bacteria at the phylum (A) and genus (B) levels; (CD) Multiple urine microbiome test results from donor Guo, stacked plots of relative abundance of urine bacteria at the phylum (C) and genus (D) levels. All urine samples were sequenced twice, and some unidentified species were removed from the results.
[0023] Figure 4 After a urine transplant, the recipient's urine microbiome resembled that of the donor. (A) After a urine transplant, recipient Ding's phylum Urobacteria levels were similar to those of donor Tang; (B) After a urine transplant, recipients Lu and Zhang's phylum Urobacteria levels were similar to those of donor Guo; (C) After a urine transplant, recipient Ding's genus Urobacteria levels were similar to those of donor Tang; (D) After a urine transplant, recipients Lu and Zhang's genus Urobacteria levels were similar to those of donor Guo. All urine samples were sequenced twice, and some unidentified species were removed from the results. DETAILED DESCRIPTION
[0024] The present invention will be further described below with reference to the embodiments of the present invention and the accompanying drawings.
[0025] Clinical data of 3 patients with IC
[0026] The patient, Ms. Ding, is 62 years old and has a history of interstitial cystitis for 6 years. She complained of lower abdominal distension and pain, unbearable urination, and the interval between urination during the day was about 20 minutes per time, and the interval between urination at night was about 30 minutes per time. She also had poor sleep and mental anxiety. Conventional treatment during the period was ineffective. On December 13, 2022, the patient underwent fecal microbiota transplantation in our hospital. She reported that the pain symptoms were slightly relieved, but relapsed soon after, and the effect was not good. Before the urinary microbiota transplantation, the patient's laboratory examination showed that the urine routine leukocyte esterase was positive, the bacterial count was slightly increased, the urine culture was normal, and other indicators were normal (Table 1); the scale assessment indicated that the patient had moderate depression, moderate anxiety, severe symptom index, severe problem index, severe genitourinary system pain, and severe pain catastrophizing (Table 2); cystoscopy showed obvious bladder mucosal inflammation ( Figure 1 A).
[0027] Patient Lu, female, 45 years old, had a history of interstitial cystitis for 3 years. She complained of frequent urination and urgency, occasional distension and pain in the lower abdomen, an interval of about 1.5h / time during daytime urination, frequent urination at night, and poor sleep quality due to nocturia. The symptoms were related to the menstrual cycle, and worsened one week after the end of the menstrual period. The disease recurred every day, and she had taken antibiotics (specific details unknown) in the past, but the effect was not good. Before the urine bacteria transplantation, the laboratory examination showed that the urine routine leukocyte esterase was positive, the bacterial count was slightly increased, the urine culture was normal, Lactobacillus jensenii was cultured in the past, and other indicators were normal (Table 1); the scale assessment showed that the patient had no anxiety or depression, mild symptom index, mild problem index, severe urogenital pain, and mild pain catastrophizing (Table 2); cystoscopy showed obvious bladder mucosal inflammation ( Figure 1 C).
[0028] The patient, Zhang Moumou, female, 79 years old, had a history of interstitial cystitis for 21 years. She complained of frequent urination and urgency accompanied by pain during urination. In severe cases, the interval between urination during the day was 15 minutes / time, the urine volume was small, 20-40mL / time, and gross hematuria was visible, and she was weak all over. She had recurrent urinary tract infections twice a month in the past, and self-administered oral antibiotics (specific details unknown) were ineffective. She had a history of hypertension. There were no obvious abnormalities in the laboratory tests before the urine bacteria transplantation, except that the bacterial count in the previous urine routine was slightly higher (Table 1); the scale assessment indicated no anxiety or depression, moderate symptom index, mild problem index, severe urogenital pain, and moderate pain catastrophizing (Table 2); cystoscopy showed obvious bladder mucosal inflammation ( Figure 1 E).
[0029] Table 1 Clinical data of 3 patients before and after urinary bacteria transplantation
[0030]
[0031]
[0032]
[0033] Table 2 Related assessment scales for 3 patients before and after urinary bacteria transplantation
[0034]
[0035] 2.3 Interstitial Cystitis Patients All Had Urine Microecological Disturbances Before Urine Bacteria Transplantation
[0036] Before urinary bacteria transplantation, we performed urine microecological tests on three patients with interstitial cystitis and found that all of them had urine microecological disorders.
[0037] The urine bacteria sequencing results before Ding Moumou's fecal microbiota transplantation were as follows: Figure 2 A (sequencing 1), Figure 2 As shown in D (sequencing 1), the phylum level of Urobacteria is dominated by Firmicutes, the genus level is dominated by Lactobacillus, and pathogenic bacteria such as Bacteroides are more abundant. Figure 2 A (sequencing 2), Figure 2 D (Sequencing 2) shows that the urine microbiome has changed compared to the previous analysis. Specifically, the Proteobacteria phylum has become dominant, the abundance of Firmicutes has decreased, the genus Lactobacillus has decreased, and the abundance of Halomonas has increased. However, pathogenic bacteria such as Pseudomonas and Ochrobacter remain at high abundance.
[0038] The results of Lu's first urine bacterial sequencing before urine bacterial transplantation were as follows: Figure 2 B (sequencing 1), Figure 2 As shown in E (sequencing 1), Firmicutes is the main phylum at the phylum level, followed by Actinobacteria. At the genus level, Lactobacillus is the main genus, followed by Gardnerella. The overall diversity of urinary bacteria is low, and pathogenic bacteria such as Gardnerella are widely present. Before the urinary bacteria transplantation, Lu was retested for urinary bacteria, and the sequencing results were as follows: Figure 2 B (sequencing 2), Figure 2 As shown in E (sequencing 2), the urinary bacteria underwent certain changes. For example, the Firmicutes at the phylum level further increased, and the corresponding Lactobacillus at the genus level also further increased. At the same time, the pathogenic bacteria Ureaplasma also increased significantly.
[0039] The results of urine bacterial sequencing before Zhang Moumou's urine bacterial transplantation are as follows Figure 2 C (sequencing 1), Figure 2As shown in F (sequencing 1), Proteobacteria dominated at the phylum level, followed by Firmicutes. At the genus level, Halomonas dominated, followed by Brevibacillus. The overall diversity of urinary bacteria was high, but opportunistic pathogens such as Cupriavidus were present.
[0040] 3. Urine bacteria transplantation
[0041] (1) Donor screening: Tang and Guo were the urine bacteria donors for the three patients in this study. Both were college students. The two donors had no abnormalities in blood routine, coagulation function and CRP tests, urine routine and urine sediment tests, stool routine and occult blood tests, related infectious disease tests, immune function tests, liver function tests, blood biochemistry and blood lipid tests, routine urine culture tests, TOUCH four tests, thyroid and parathyroid tests, sex hormone combination tests, tumor marker screening and other related tests. EQUC test showed no pathogenic bacteria contamination. The 16s rRNA sequencing of the donor urine bacteria showed a high diversity of urine bacteria ( Figure 3 The results of multiple urine bacterial sequencing at different time points indicate that the donor urine microecology is relatively stable ( Figure 3 The urine bacteria of both groups are dominated by probiotics such as Bacillus, Halomonas, and Streptococcus, while pathogenic bacteria such as Escherichia and Proteus are relatively rare, making them excellent donors for urine bacteria transplantation.
[0042] (2) Urine microbial transplantation can significantly adjust the patient's urine microecology compared to fecal microbial transplantation
[0043] Ding Moumou underwent fecal microbiota transplantation on December 13, 2022, and reported poor efficacy after the operation. The urine bacteria sequencing results of Ding Moumou before and after fecal microbiota transplantation were as follows: Figure 2 A (Urine bacteria sequencing results before fecal microbiota transplantation: Sequencing 1; Urine bacteria sequencing results after fecal microbiota transplantation: Sequencing 2), Figure 2 Figure D (Urine bacteria sequencing results before fecal microbiota transplantation: Sequencing 1; urine bacteria sequencing results after fecal microbiota transplantation: Sequencing 2). The results suggest that the patient's urine microbiome changed only slightly, and the abundance of pathogenic bacteria such as Pseudomonas and Ochrobacter remained high.
[0044] On March 11, 2023, Ding Moumou underwent urine bacteria transplantation in our hospital (urine bacteria donor: Tang Mou). Seven days after the urine bacteria transplantation, the patient reported that the symptoms of frequent urination improved and the pain of urination was relieved. Laboratory tests showed no abnormalities (Table 1). The urine microecology was retested 2 weeks after the urine bacteria transplantation, and the results showed significant changes compared with the fecal bacteria transplantation. The specific results are as follows Figure 2 A (sequencing three), Figure 2D (Sequencing 3). At the Urobacteria level, Proteobacteria decreased slightly but remained the primary component, while Firmicutes decreased further. Actinobacteria and Fusobacteria increased in abundance. At the genus level, Lactobacillus decreased further, while Gardnerella and Dietzia increased. Meanwhile, Halomonas, a probiotic bacterium with high abundance in donors, also increased.
[0045] After Ding Moumou underwent urine bacteria transplantation, his urine microecology was similar to that of the donor Tang Mou. The results were as follows: Figure 4 A. Figure 4 As shown in C.
[0046] (3) Urine bacteria transplantation is safe and effective
[0047] On June 16, 2022, Lu Mou underwent urine bacteria transplantation in our hospital (urine bacteria donor: Guo Moumou). On the day after the transplantation, the patient's lower abdominal distension and pain improved, and urination was smooth; no adverse reactions such as fever and allergies occurred within 24 hours after the operation; one week after the transplantation, the symptoms of frequent urination and urgency were alleviated, and he occasionally got up at night. Laboratory tests showed no abnormalities (Table 1). Nine months after the urine bacteria transplantation, the urine microecology was retested, and the results were as follows Figure 2 B (sequencing three), Figure 2 As shown in E (sequencing 3), Firmicutes is still the dominant genus at the phylum level, and Streptococcus has increased significantly and become the dominant genus. After Lu Mou underwent urine bacterial transplantation, his urine microecology was similar to that of the donor Guo Moumou. The results are as follows Figure 4 B. Figure 4 As shown in D.
[0048] On March 19, 2023, Zhang Moumou underwent urine bacteria transplantation in our hospital (urine bacteria donor: Guo Moumou). Symptoms were relieved on the day after the transplantation, sleep was good, and mood was good; no adverse reactions such as fever and allergies occurred within 24 hours after the operation; one week later, the symptoms of frequent urination and urgency were basically improved, and he urinated about once every 3 hours during the day, and the urine volume increased. Laboratory tests showed no abnormalities (Table 1). Urine bacteria were collected the day after the urine bacteria transplantation, and the sequencing results were as follows Figure 2 C (sequencing 2), Figure 2 As shown in F (sequencing 2), at the phylum level, Firmicutes increased and became the dominant bacterial group, Proteobacteria decreased significantly, Actinobacteria increased slightly, and at the genus level, Streptococcus increased significantly compared to before the urine bacteria transplantation, Bacteroides increased slightly, and Cupriavidus was not seen. After the urine bacteria transplantation, the diversity of urine bacteria in patients increased and opportunistic pathogens decreased. After the urine bacteria transplantation, Zhang Moumou's urine microecology was close to that of the donor Guo Moumou. The results are as follows Figure 4 B. Figure 4 As shown in D.
[0049] (4) Follow-up and prognosis
[0050] We conducted long-term follow-up on 3 patients after urinary bacteria transplantation.
[0051] On May 16, 2023, more than two months after the urine bacteria transplantation, Ding Moumou's relevant laboratory tests were reviewed and no abnormalities were found (Table 1); the scores of the relevant scales decreased, indicating that anxiety and depression improved, symptoms were alleviated, and the impact on quality of life was reduced (Table 2); a cystoscopy was performed simultaneously ( Figure 1 B) Scattered mucosal bleeding and inflammation were observed, indicating improvement compared to before the urinary bacterial transplant. During a 15-month follow-up, the patient's lower abdominal pain and dysuria disappeared, and there was no recurrence of frequent urination.
[0052] On March 29, 2023, more than 9 months after the urinary bacteria transplantation, Lu's laboratory tests were normal (Table 1); the scores on the relevant scales decreased, indicating that his physical and mental conditions had improved (Table 2); cystoscopy showed scattered inflammation of the mucosa, but no Hunner's ulcer was found, which was better than before the urinary bacteria transplantation ( Figure 1 D). During the subsequent 20-month follow-up, a urine routine examination on April 14, 2023, revealed an increase in white blood cells and leukocyte esterase. A urine routine examination on May 6, 2023, revealed an increase in the number of red blood cells, white blood cells, leukocyte esterase, and bacteria. Urine culture was positive for Streptococcus lactis. Other abnormalities were normal, and the patient did not complain of recurrence of lower urinary tract symptoms.
[0053] On May 12, 2024, more than 13 months after the urinary bacteria transplantation, a urine routine examination showed an increase in white blood cells and leukocyte esterase, with no other abnormalities (Table 1); the relevant scale score decreased, indicating that both physical and mental conditions had improved (Table 2); cystoscopy showed signs of mucosal inflammation, which was better than before the urinary bacteria transplantation ( Figure 1 F) After 26 months of follow-up, no recurrence was observed and the lower urinary tract symptoms were completely improved.
Claims
1. A urinary bacteria transplant solution for treating interstitial cystitis, characterized in that: The invention is composed of the following components: urine sediment obtained by centrifugation of urine from a healthy donor, and sterile physiological saline. The concentration of the urine sediment in the urine transplant solution is 0.01-0.05 g / L.
2. The urinary bacteria transplantation solution for treating interstitial cystitis according to claim 1, characterized in that: The abundance of any one of the genus Lactobacillus, Corynebacterium, Prevotella, Staphylococcus, and Streptococcus in the urine bacterial transplant solution should be in the top ten of the abundance ranking of all bacterial species.
3. A method for preparing a urine bacteria transplant solution for treating interstitial cystitis, characterized in that: The main steps are as follows:
1. Donor screening. Donors are between 18 and 22 years old and have a healthy diet and lifestyle. All donors need to undergo basic health examinations. Donors who pass the basic health examinations will undergo further urine microecological testing; 2. Urine collection: Aseptic catheterization was used to drain 150 ml of urine into a sterile container and temporarily stored at 4°C.
3. Urine bacteria extraction: 1) Centrifuge 150 ml of urine to separate the supernatant and urine bacteria precipitate, and remove the supernatant; 2) In a laminar flow hood, resuspend the urine bacterial pellet in 40 ml of sterile saline to prepare a urine bacterial transplant solution.
4. The method for preparing a urine bacteria transplant solution for treating interstitial cystitis according to claim 3, characterized in that: In step 1, urine microecological testing also includes 16S rRNA sequencing analysis of urine bacteria in donor urine specimens. The relevant primer pair sequence is F: GTGCCAGCMGCCGCGGTAA, R: GGACTACHVGGGTWTCTAAT. Donors whose urine flora abundance of any one of the genera Lactobacillus, Corynebacterium, Prevotella, Staphylococcus, or Streptococcus is in the top ten of the total species abundance are selected.
5. Use of the urine bacteria transplant solution for treating interstitial cystitis as claimed in claim 1 in a drug for treating refractory interstitial cystitis.