The association between vaginal microbiota disorders and early missed abortion: A prospective study.

INTRODUCTION: The objective of this study was to explore the association between disordered vaginal microbiota and missed abortion to ascertain potential causes of missed abortion related to vaginal microbiology. MATERIAL AND METHODS: This study was a prospective cohort study with a comparison group (reference group). Vaginal secretions from the posterior fornix of women in the early stages of pregnancy were collected in the Xiangya Third Hospital of Central South University in Changsha, Hunan, China, from November 2018 to November 2019. A total of 54 cases of missed abortion (case group) and 50 cases of normal pregnancy requiring induced abortion (reference group) were analyzed. Bacterial DNA was extracted, hybridized with gene-specific primers, and then detected using a bacterial chip. The composition and relative abundance of vaginal microbiota in the two groups were compared using α-diversity analysis, ß-diversity analysis, and the linear discriminant analysis effect size method. RESULTS: The α-diversity analysis showed that the Simpson index of the case group was lower than that of the reference group, whereas the Shannon index in the case group was higher. The relative abundance of Firmicutes in the case group (42.52%) was lower than in the reference group (51.03%, p < 0.05), as was the relative abundance of Lactobacillus (case group 16.51%, reference group 23.00%; p < 0.05). Interestingly, levels of Mycoplasma genitalium and Ureaplasma were lower in the case group (p < 0.05). The relative abundance of Lactobacillus crispatus, Lactobacillus jensenii, and Lactobacillus gasseri was also significantly lower in the case group than in the reference group (p < 0.05). The pathways enriched in the case group were predominantly related to metabolism, whereas the genetics-related pathways were predominantly enriched in the reference group. CONCLUSIONS: Bacteria are more diverse and unevenly distributed in patients with missed abortion. Decreases in the proportion of vaginal Lactobacillus and changes in Lactobacillus species in these patients may increase the chance of genital tract pathogenic bacterial infection. To our knowledge, our study was the first to observe that a decrease of Firmicutes levels in the vaginal microbiota might impair energy metabolism and have an association with missed abortion.

The density of endometrial glandular openings: a novel variable to predict the live birth rate in patients with intrauterine adhesions following hysteroscopic adhesiolysis.

STUDY QUESTION: Can the density of endometrial glandular openings (DEGO) be a reliable and simple new variable in the prediction of live birth after hysteroscopic adhesiolysis? SUMMARY ANSWER: The DEGO grade at follow-up hysteroscopy outperforms American Fertility Society (AFS) score in predicting the live birth rate after hysteroscopic adhesiolysis for patients with intrauterine adhesions (IUAs). WHAT IS KNOWN ALREADY: Several methods, such as endometrial thickness and AFS score, have been proposed for predicting the live birth rate in patients with IUAs who undergo hysteroscopic adhesiolysis. STUDY DESIGN, SIZE, DURATION: A test cohort of 457 patients with IUAs who underwent hysteroscopic adhesiolysis and had satisfactory follow-up hysteroscopy videos were retrospectively enrolled between January 2016 and January 2017. A validation cohort comprising 285 IUA patients was prospectively enrolled from March 2018 to August 2018. PARTICIPANTS/MATERIALS, SETTING, METHODS: An automated counting software tested the follow-up hysteroscopy videos to calculate the DEGO grade of all the 742 patients with IUAs after hysteroscopic adhesiolysis. The AFS score for each patient was also calculated at the same follow-up hysteroscopy. Logistic regression analysis was performed to develop prediction models to predict the live birth rate following hysteroscopic adhesiolysis. The performance of each of these prediction models was compared by calculating the AUC. MAIN RESULTS AND THE ROLE OF CHANCE: In the test cohort (n = 457), 231 patients had a live birth, but 226 patients failed. In the validation cohort (n = 285), 117 patients had a live birth, while 168 patients did not. The logistic regression analysis revealed that both the DEGO grade and AFS score at follow-up hysteroscopy were closely correlated with the live birth rate in patients with IUAs (P = 0). The AUCs of AFS score and DEGO grade in the test cohort were 0.7112 and 0.8498, respectively (P < 0.0001). The AUCs of AFS score and DEGO grade in the prospective external validation cohort were 0.6937 and 0.8248, respectively (P < 0.0001). LIMITATIONS, REASONS FOR CAUTION: Further well-designed prospective clinical studies with a multicentric larger sample size should be needed to confirm the feasibility and efficacy of DEGO. WIDER IMPLICATIONS OF THE FINDINGS: The DEGO grade is an accurate predictor factor of live birth rate in patients with IUAs following hysteroscopic adhesiolysis and can represent in the future an important and promising tool for assessing obstetric outcomes in IUAs. STUDY FUNDING/COMPETING INTEREST(S): This study is supported by National Key Research and Development Program of China (Grant No. 2018YFC1004800), Natural Science Foundation of China (Grant No. 81671492), Natural Science Foundation of Hunan (Grant No. 2020JJ5859). B.G. is supported by Chinese Scholarship Council (File number. 201806370178). The authors have no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Comparison of ultrasound-guided high-intensity focused ultrasound ablation and hysteroscopic myomectomy for submucosal fibroids: a retrospective study.

OBJECTIVE: To compare the safety, reintervention and pregnancy outcomes between ultrasound-guided high intensity focused ultrasound (USgHIFU) and hysteroscopic myomectomy (HM) for submucosal fibroids. MATERIALS AND METHODS: A total of 215 patients with a solitary submucosal fibroid treated by USgHIFU or HM at the third Xiangya Hospital were retrospectively reviewed. Among them, 58 treated with USgHIFU, 157 treated with HM. RESULTS: A significant difference was observed in size, location and type of the fibroids, effective rate, and cumulative reintervention rate between the two groups (p < .05). The size of the fibroids was 57.9 ± 1.9 mm in the USgHIFU group, while it was 32.6 ± 1.2 mm in the HM group. The number of the fibroids at horn or fundus/uterine cavity was 16/42 in the USgHIFU group, while it was 21/136 in the HM group. The number of type I/II/2-5 was 16/17/25 in the USgHIFU group, while it was 133/24/0 in the HM group. In the USgHIFU group, the effective rate was 100% and the cumulative reintervention rate at 50 (17-97) months was 19.0%, while in the HM group, it was 94.3% and 7.6%, respectively. During the follow-up period, the pregnancy rate was 22.4% (13/58) and the reintervention rate due to invalid and recurrence was 15.5% (9/58) in the USgHIFU group, while they were 18.5% (29/157) and 7.0% (11/157) in the HM group. No significant difference was observed between the two groups (p > .05). Furthermore, the reintervention rate was positively correlated with age, treatment methods and parity and fertility requirements. No other significant difference was observed between the two groups. CONCLUSIONS: Both USgHIFU and HM are safe and effective in treating submucosal fibroids. Compared with the HM group, the USgHIFU group had lower postoperative complications, but higher reintervention rate, with similar recurrence rate, pregnancy rate and reintervention rate due to invalid and recurrence. Reintervention was related to age, treatment methods, parity and fertility requirements.

The impact of serum incubation time on IgM/IgG binding to porcine aortic endothelial cells.

The results of the assay for measuring anti-non-Gal antibodies (which affect pig xenograft survival) in recipients are important. Serum incubation time and concentration may be important factors in the extent of antibody binding to the graft. The aim of this in vitro study was to determine the optimal incubation time and serum concentration for measuring anti-non-Gal antibody binding to porcine aortic endothelial cells (pAECs). Pooled human, naive, and sensitized baboon sera were incubated with wild-type, α1,3-galactosyltransferase gene-knockout (GTKO), and GTKO/human CD55 pAECs. IgM/IgG binding to pAECs after varying serum incubation times (0.5, 1, 2, and 3 hour) and concentrations (5, 10, 20, and 40 µL) was determined by flow cytometry. An increase in incubation time from 30 minutes to 2 hour was associated with increases in anti-non-Gal IgM/IgG binding to GTKO and GTKO/hCD55 pAECs of pooled human, naive and sensitized baboon sera (P<.05). Pooled human serum showed a significant increase in anti-non-Gal IgM (1.5 times) and a minimal increase in anti-non-Gal IgG antibody binding. IgM/IgG binding of sensitized baboon serum to GTKO pAECs after 2-hour incubation was 1.5 times and 2 times greater than after 30-minutes incubation, respectively, whereas naïve baboon sera showed minimal (non-significant) increase in anti-non-Gal IgM/IgG antibody binding. With 2-hour incubation, increasing the serum concentration from 5 µL to 20 µL significantly increased antibody binding to non-Gal antigens in pooled human and sensitized baboon serum. With naïve baboon serum, only IgG was significantly increased. Increasing the serum incubation time contributed to improve the sensitivity of detecting anti-non-Gal antibodies, without affecting cell viability in vitro.

Therapeutic regulation of systemic inflammation in xenograft recipients.

Inflammation is known to preclude tolerance after transplantation. We have previously shown that systemic inflammation in xenograft recipients (SIXR) precedes activation of coagulation in the absence of T cell responses. Accordingly, SIXR may amplify innate and adaptive immune responses against xenografts after pig-to-primate xenotransplantation, even with efficient immunosuppressive therapy. We evaluated the impact of anti-inflammatory agents on pro-inflammatory cytokines and chemokines in pig artery patch and heart xenograft recipients. Baboons received an artery patch (Group1, n=8) or heart (Group2, n=4) from genetically engineered pigs. All baboons received lymphodepletion with thymoglobulin (ATG) and costimulation blockade-based immunosuppression (anti-CD40 and/or CTLA4Ig). In Group1, baboons received either (i) no anti-inflammatory agents (n=2), (ii) cobra venom factor (CVF, n=2), (iii) α1-antitrypsin (AAT, n=2), or (iv) interleukin (IL)-6 receptor antagonist (IL-6RA, n=2). In Group2, all baboon received corticosteroids, either without (n=2) or with (n=2) IL-6RA. Serum IFN-γ, TNF-α, IL-1ß, IL-17, IL-6, IL-8, MCP-1, and sCD40L levels were measured by Luminex. Fibrinogen, D-dimers, and C-reactive protein (C-RP) were also measured. Recipient baboon T cell proliferation was evaluated by mixed lymphocyte reaction (MLR) before and after transplantation. Pig and baboon tissue factor (TF) mRNA levels in heart xenografts were measured by RT-PCR. In no recipient was a marked increase in T cell response to pig cells observed after transplantation. In Groups 1 and 2, post-transplantation levels of IFN-γ, TNF-α, IL-1ß, and IL-17 remained comparable to or lower than pre-transplant levels, except in one heart recipient that succumbed to CMV infection. In Group1, when no anti-inflammatory agent was administered, post-transplant levels of IL-6, IL-8, and MCP-1 were elevated. After CVF, IL-6, IL-8, and MCP-1 remained low. After IL-6RA, IL-6 and MCP-1 were elevated. After AAT, IL-8 was elevated. sCD40L became elevated intermittently in most recipients irrespective of the administered anti-inflammatory agent. In Group2, IL-6 was transiently elevated, particularly after IL-6RA administration. MCP-1 gradually increased by 2 months in Group2 recipients. sCD40L generally remained low except in one recipient. In Group1 and Group2 recipients, C-RP levels were elevated except after IL-6RA administration, while D-dimers were elevated regardless of administration of anti-inflammatory agent. In Group2, pig TF mRNA levels were increased in heart xenografts compared to naive pig hearts, irrespective of IL-6 receptor antagonist administration. Additionally, baboon TF mRNA levels were detectable in heart xenografts, but not in naive pig hearts. Some pro-inflammatory cytokines and chemokines are elevated in xenograft recipients, even with efficient T cell-directed immunosuppressive therapy. Persistent elevation of D-dimers, and individual cytokines and chemokines suggest a continuous inflammatory response, despite administration of anti-inflammatory agents. Systemic administration of combined anti-inflammatory agents as well as complement regulation may be essential to prevent SIXR after xenotransplantation.

Immunological and physiological observations in baboons with life-supporting genetically engineered pig kidney grafts.

BACKGROUND: Genetically engineered pigs could provide a source of kidneys for clinical transplantation. The two longest kidney graft survivals reported to date have been 136 and 310 days, but graft survival >30 days has been unusual until recently. METHODS: Donor pigs (n=4) were on an α1,3-galactosyltransferase gene-knockout (GTKO)/human complement regulatory protein (CD46) background (GTKO/CD46). In addition, the pigs were transgenic for at least one human coagulation regulatory protein. Two baboons received a kidney from a six-gene pig (GroupA) and two from a three-gene pig (GroupB). Immunosuppressive therapy was identical in all four cases and consisted of anti-thymoglobulin (ATG)+anti-CD20mAb (induction) and anti-CD40mAb+rapamycin+corticosteroids (maintenance). Anti-TNF-α and anti-IL-6R mAbs were administered to reduce the inflammatory response. Baboons were followed by clinical/laboratory monitoring of immune/coagulation/inflammatory/physiological parameters. At biopsy or euthanasia, the grafts were examined by microscopy. RESULTS: The two GroupA baboons remained healthy with normal renal function >7 and >8 months, respectively, but then developed infectious complications. However, no features of a consumptive coagulopathy, eg, thrombocytopenia and reduction of fibrinogen, or of a protein-losing nephropathy were observed. There was no evidence of an elicited anti-pig antibody response, and histology of biopsies taken at approximately 4, 6, and 7 months and at necropsy showed no significant abnormalities. In contrast, both GroupB baboons developed features of a consumptive coagulopathy and required euthanasia on day 12. CONCLUSIONS: The combination of (i) a graft from a specific six-gene genetically modified pig, (ii) an effective immunosuppressive regimen, and (iii) anti-inflammatory therapy prevented immune injury, a protein-losing nephropathy, and coagulation dysfunction for >7 months. Although the number of experiments is very limited, our impression is that expression of human endothelial protein C receptor (±CD55) in the graft is important if coagulation dysregulation is to be avoided.

Identification of Alternative Splicing-Related Genes CYB561 and FOLH1 in the Tumor-Immune Microenvironment for Endometrial Cancer Based on TCGA Data Analysis.

Background: Advanced and recurrent endometrial cancer EC remains controversial. Immunotherapy will play a landmark role in cancer treatment, and alternative splicing (AS) of messenger RNA (mRNA) may offer the potential of a broadened target space. Methods: We downloaded the clinical information and mRNA expression profiles from The Cancer Genome Atlas (TCGA) database. Hub genes were extracted from 11 AS-related genes to analyze the correlation between clinical parameters and the tumor-immune microenvironment. We also analyzed the correlations between the copy numbers, gene expressions of hub genes, and immune cells. The correlation between the risk score and the six most important checkpoint genes was also investigated. The ESTIMATE algorithm was finally performed on each EC sample based on the high- and low-risk groups. Results: The risk score was a reliable and stable independent risk predictor in the Uterine Corpus Endometrial Carcinoma (UCEC) cohort. CYB561|42921|AP and FOLH1|15817|ES were extracted. The expression of CYB561 and FOLH1 decreased gradually with the increased grade and International Federation of Gynecology and Obstetrics (FIGO) stage (p < 0.05). Gene copy number changes in CYB561 and FOLH1 led to the deletion number of myeloid DC cells and T cell CD8+. Low expression of both CYB561 and FOLH1 was associated with poor prognosis (p < 0.001). The checkpoint genes, CTLA-4 and PDCD1, exhibited a negative correlation with the risk score of AS in UCEC. Conclusion: AS-related gene signatures were related to the immune-tumor microenvironment and prognosis. These outcomes were significant for studying EC's immune-related mechanisms and exploring novel prognostic predictors and precise therapy methods.

Pregnancy Patterns Impact Live Birth Rate for Patients With Intrauterine Adhesions After Hysteroscopic Adhesiolysis: A Retrospective Cohort Study.

Objective: The pregnancy patterns and other factors of live birth for patients with intrauterine adhesions (IUAs) were identified by analyzing the clinical features of pre-, intra-, and post-hysteroscopic adhesiolysis (HA). Design: A total of 742 patients with IUAs who wanted to become pregnant underwent HA from January 2017 to May 2018 at the Third Xiangya Hospital of Central South University. The patient follow-up period was 2 years post-HA. A logistic regression was performed to analyze the clinical characteristics associated with a live birth for patients with IUAs. Pre-operative clinical indicators included age, gravidity, parity, abortion, IUA recurrence, menstrual patterns, disease course. Intraoperative clinical features assessed in the last operation were uterine cavity length, IUA appearance, IUA area, number of visible uterine cornua, number of visible tubal ostia, AFS scores. Pregnancy patterns were post-hysteroscopic adhesiolysis features. Results: Among the 742 IUA patients, 348 (46.9%) had a live birth and 394 (53.1%) did not. A bivariate and binary logistic regression analysis showed that IUA patients' pregnancy patterns, age, number of visible tubal ostia noted by a second-look hysteroscopy, and American Fertility Society (AFS) scores were significantly related to the live birth rate (P < 0.05). Conclusions: Pregnancy patterns, age, number of visible tubal ostia, and AFS scores were significantly related to the live birth rate and may be considered potential predictors of the live birth rate in IUA patients. The indications of assisted reproductive technology (ART) might be a better choice for patients with recurrent IUAs.

Characteristics and transcriptomic analysis of scar tissues on the inner uterine cavity wall in patients with intrauterine adhesions.

Introduction: It has been previously reported that intrauterine adhesions (IUAs) are the main cause of uterine infertility. However, the histological origin of scar tissue present on the inner wall of the uterine cavity with IUAs has not been previously studied, which is particularly necessary for follow-up research and prevention and treatment. Methods: In this study, myometrium with normal uterus were assigned to the control group and scar tissues with IUAs were assigned to the experimental group. And pathological characteristics and transcriptomic were analyzed between the two groups. Results: We founded no difference was noted in the histological morphology and the α-SMA expression between the experimental and control groups. A total of 698 differentially expressed genes were identified between the two groups. Gene Ontology (GO) analyses revealed that the DEGs were significantly enriched in cell proliferation, AP-1 complex formation, and angiogenesis. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses revealed that the target genes were significantly enriched in the AGE-RAGE, FOXO and TNF signaling pathway. Discussion: As far as we know, this is the first study to propose that the scar tissues are mainly derived from the myometrium and the first one to report differentially expressed genes in the scar tissues of IUAs.

A nomogram model based on clinical markers for predicting malignancy of ovarian tumors.

Objective: The aim of this study was to build a nomogram based on clinical markers for predicting the malignancy of ovarian tumors (OTs). Method: A total of 1,268 patients diagnosed with OTs that were surgically removed between October 2017 and May 2019 were enrolled. Clinical markers such as post-menopausal status, body mass index (BMI), serum human epididymis protein 4 (HE4) value, cancer antigen 125 (CA125) value, Risk of Ovarian Malignancy Algorithm (ROMA) index, course of disease, patient-generated subjective global assessment (PG-SGA) score, ascites, and locations and features of masses were recorded and analyzed (p 0.05). Significant variables were further selected using multivariate logistic regression analysis and were included in the decision curve analysis (DCA) used to assess the value of the nomogram model for predicting OT malignancy. Result: The significant variables included post-menopausal status, BMI, HE4 value, CA125 value, ROMA index, course of disease, PG-SGA score, ascites, and features and locations of masses (p 0.05). The ROMA index, BMI (≥ 26), unclear/blurred mass boundary (on magnetic resonance imaging [MRI]/computed tomography [CT]), mass detection (on MRI/CT), and mass size and features (on type B ultrasound [BUS]) were screened out for multivariate logistic regression analysis to assess the value of the nomogram model for predicting OT malignant risk (p 0.05). The DCA revealed that the net benefit of the nomogram's calculation model was superior to that of the CA125 value, HE4 value, and ROMA index for predicting OT malignancy. Conclusion: We successfully tailored a nomogram model based on selected clinical markers which showed superior prognostic predictive accuracy compared with the use of the CA125, HE4, or ROMA index (that combines both HE and CA125 values) for predicting the malignancy of OT patients.

Modified HE4, CA125, and ROMA cut-off values and predicted probability of ovarian tumor in Chinese patients.

BACKGROUND: Most prior studies investigating the risk of ovarian malignancy algorithm (ROMA) with cancer antigen 125 (CA125) and human epididymis protein 4 (HE4) have involved Caucasian population or other populations. To date, there have been no unique calculations of predicted probability (PP) risk specifically for Chinese populations to help physicians in primary care settings. METHODS: A group of 534 women with ovarian tumor diagnoses were enrolled and serum HE4 and CA125 were measured in each individual. Modified cut-off values were obtained by maximizing area under the curve (AUC) values and adjusted by using logistic regression with corresponding sensitivity (SN), specificity (SP), Youden index (YI), positive predictive value (PPV), and negative predictive value (NPV). RESULTS: By utilizing the ideal PPV, NPV, and AUC values, in premenopausal women modified HE4, CA125, ROMA, and PP cut-off values were 73.87 pmol/L, 61.60 U/mL, 18.47%, and 0.168, respectively. The same test values for postmenopausal women were 120.90 pmol/L, 76.21 U/mL, 26.48%, and 0.485, respectively. The SN for HE4 with the modified cut-off value was significantly lower than that for CA125 (P=0.040) in premenopausal women and lower than that for ROMA (P=0.001) and PP (P=0.044) in postmenopausal women. The AUC values for CA125, ROMA, and PP were all significantly higher than that for HE4 (P=0.006, 0.007, and 0.002, respectively) in postmenopausal women. CONCLUSIONS: The modified cut-off values for HE4, CA125, ROMA, and PP with ideal SN, SP, YI, NPV, PPV were useful of ruling out ovarian malignancy among both pre- and post-menopausal women. In premenopausal women modified HE4, CA125, ROMA, and PP cut-off values were 73.87 pmol/L, 61.60 U/mL, 18.47%, and 0.168, respectively and in postmenopausal women were 120.90 pmol/L, 76.21 U/mL, 26.48%, and 0.485, respectively.

The characteristics and risk factors of human papillomavirus infection: an outpatient population-based study in Changsha, Hunan.

This cross-sectional study investigated the characteristics of cervical HPV infection in Changsha area and explored the influence of Candida vaginitis on this infection. From 11 August 2017 to 11 September 2018, 12,628 outpatient participants ranged from 19 to 84 years old were enrolled and analyzed. HPV DNA was amplified and tested by HPV GenoArray Test Kit. The vaginal ecology was detected by microscopic and biochemistry examinations. The diagnosis of Candida vaginitis was based on microscopic examination (spores, and/or hypha) and biochemical testing (galactosidase) for vaginal discharge by experts. Statistical analyses were performed using SAS 9.4. Continuous and categorical variables were analyzed by t-tests and by Chi-square tests, respectively. HPV infection risk factors were analyzed using multivariate logistic regression. Of the total number of participants, 1753 were infected with HPV (13.88%). Females aged ≥ 40 to < 50 years constituted the largest population of HPV-infected females (31.26%). The top 5 HPV subtypes affecting this population of 1753 infected females were the following: HPV-52 (28.01%), HPV-58 (14.83%), CP8304 (11.47%), HPV-53 (10.84%), and HPV-39 (9.64%). Age (OR 1.01; 95% CI 1-1.01; P < 0.05) and alcohol consumption (OR 1.30; 95% CI 1.09-1.56; P < 0.01) were found to be risk factors for HPV infection. However, the presence of Candida in the vaginal flora was found to be a protective factor against HPV infection (OR 0.62; 95% CI 0.48-0.8; P < 0.001). Comparing with our previous study of 2016, we conclude that the subtype distribution of HPV infection is relatively constant in Changsha. Our data suggest a negative correlation between vaginal Candida and HPV, however, more radical HPV management is required in this area for perimenopausal women and those who regularly consume alcohol.

Imbalance of the Gut Microbiota May Be Associated with Missed Abortions: A Perspective Study from a General Hospital of Hunan Province.

OBJECTIVE: To conduct a preliminary investigation that shows the possible correlation between the change of gut microbiota and missed abortions (MAs), which further provides a new potential insight for the prevention and therapy of MAs. METHOD: One hundred women, including 50 patients with MAs (case group) and 50 normal pregnant women (control group), were enrolled in the study. Fecal specimens were collected in the first trimester. Bacterial DNA was extracted, hybridized with primers of specific genes, and then detected by bacterial chip. The composition and the relative abundance of the gut microbiota were compared and analyzed. Furthermore, Kyoto Encyclopedia of Genes and Genomes enrichment analysis was used to explore the relative pathways. RESULTS: (1) The α-diversity and ß-diversity of the gut microbiota in patients with MAs were significantly lower than that those in normal pregnant women (P < 0.05). At the phylum level, Firmicutes, Proteobacteria, Actinomycetes, and Bacteroidetes accounted for the main proportion of intestinal flora in the 2 groups. Only Actinobacteria was high in the case group. Significant differences were found between the two groups at the phylum level (P < 0.05). Prevotella, Lactobacillus, and Paracoccus were significantly more abundant in the control group than in the case group at the genus level (P < 0.05). (2) KEGG pathway enrichment analysis found significant differences in 27 signaling pathways and metabolic pathways between the two groups of differentially expressed genes (all adjusted P < 0.05). (3) The positive rate of M. hominins (MH) detection in the control group was significantly higher in the MA group (χ 2 = 7.853, P = 0.004). CONCLUSION: The high abundance of Actinobacteria in the MA group was the first time found and reported in the study. The dysbiosis of the gut microbiota correlates with MAs. This study provided insights into the potential change of gut microbiota of MAs and the potential underlying mechanisms through certain impaired lipid metabolism and aroused inflammation pathways. Comprehensive insights regarding gut microbiota may facilitate improved understanding and the development of novel therapeutic and preventive strategies for MAs.

Hysteroscopic incision of the incomplete uterine septum using 5-French scissors with marking strategies: a modified hysteroscopic technique.

OBJECTIVE: To demonstrate an easier surgical strategy by using the marking technique for hysteroscopic incision of the uterine septum using 5-French cold scissors. DESIGN: A step-by-step surgical video demonstration. SETTING: Gynecologic department of the affiliated hospital. PATIENT(S): A 33-year-old woman presented with a 4-year history of primary infertility. She previously had undergone transcervical resection of (uterine) septum owing to the presence of a complete uterine septum and double cervices. Postoperative 3-dimensional ultrasound revealed a 1.2-cm residual uterine septum, and the outline of the uterine fundus was flat. A second surgery for resection of the residual septum was recommended before in vitro fertilization and embryo transfer. We used the Campo hysteroscope (4.4-mm outer sheath) and 5-French scissors with our modified marking strategy to incise the incomplete uterine septum. INTERVENTION(S): There were several critical strategies for this approach. After fully exposing 2 fallopian tube ostia, a 3-5-mm mark was made on each side of the uterine fundus where the septum ended, and the marks were parallel to the fallopian tubal ostia. The septum then was incised along the line between the two previously marked points that served as the endpoints. Care was taken to avoid incising myometrial blood vessels during incision, and the 5-French bipolar electrode was used for coagulation when necessary. At the end of the surgery, the distension pressure was gradually decreased to 80 mm Hg to confirm hemostasis of the wound before withdrawing the hysteroscope. MAIN OUTCOME MEASURE(S): Description of a modified hysteroscopic technique. RESULT(S): The overall operation time was 10 minutes, and the estimated blood loss was 5 mL. The residual septum was resected successfully while maintaining optimal hysteroscopic visualization. There were no short-term complications, such as uterine perforation or fluid overload. Hysteroscopic evaluation performed 3 months after surgery revealed that the uterine cavity was nearly normal, with no intrauterine adhesion appreciated. There are several advantages to this innovative and practical hysteroscopic surgical approach. Marking the lateral limits of the uterine septum means that a shorter reference line is obtained to incise the septum effectively rather than using the bilateral ostia as reference points. At the same time, marking the bilateral endpoint of the uterine septum incision at the beginning of the surgery might be helpful when bilateral tubal ostia are invisible because of quick absorption of the distension media, which causes insufficient distention pressure at the end of the surgery. Use of the narrow 5-French scissors allowed for instrumentation without prior cervical dilation. Moreover, with this "see and treat" strategy, a clear visualization of the surgical field was maintained without inserting and withdrawing the hysteroscope. The endometrium sustained minimal damage because of the "cold scissors" technique. CONCLUSION(S): Our hysteroscopic marking strategy allows the surgeon's intraoperative judgment to be efficient and safe during incision of the uterine septum and ensures that the incision is adequate. It is an improved and valid surgical strategy for hysteroscopic incision of the uterine septum.

Cancer-associated MSC drive tumor immune exclusion and resistance to immunotherapy, which can be overcome by Hedgehog inhibition.

We investigated the impact of cancer-associated mesenchymal stem cells (CA-MSCs) on ovarian tumor immunity. In patient samples, CA-MSC presence inversely correlates with the presence of intratumoral CD8+ T cells. Using an immune "hot" mouse ovarian cancer model, we found that CA-MSCs drive CD8+ T cell tumor immune exclusion and reduce response to anti­PD-L1 immune checkpoint inhibitor (ICI) via secretion of numerous chemokines (Ccl2, Cx3cl1, and Tgf-ß1), which recruit immune-suppressive CD14+Ly6C+Cx3cr1+ monocytic cells and polarize macrophages to an immune suppressive Ccr2hiF4/80+Cx3cr1+CD206+ phenotype. Both monocytes and macrophages express high levels of transforming growth factor ß­induced (Tgfbi) protein, which suppresses NK cell activity. Hedgehog inhibitor (HHi) therapy reversed CA-MSC effects, reducing myeloid cell presence and expression of Tgfbi, increasing intratumoral NK cell numbers, and restoring response to ICI therapy. Thus, CA-MSCs regulate antitumor immunity, and CA-MSC hedgehog signaling is an important target for cancer immunotherapy.

Cold scissors ploughing technique in hysteroscopic adhesiolysis: a comparative study.

BACKGROUND: Intrauterine adhesions (IUAs) can be dissected using hysteroscopic scissors (cold scissors) or other methods, but there is no consensus on which hysteroscopic method is preferable. There is also no consensus on the method of how to deal with the scar tissue on the surface of the intrauterine cavity. METHODS: From January 2016 to October 2017, 179 patients who had HA met the enrollment criteria (see the text below), and their data were analyzed retrospectively. In addition, all patients were divided into three groups according to the surgical techniques used. The groups were the ploughing group (PG) (using cold scissors to dissect the adhesion and cut the scar tissue using a ploughing technique) (n=81), the traditional group (TG) (using cold scissors to dissect the adhesion, but not deal with the scar tissue) (n=42), and the electrosurgical group (EG) (using a resectoscope to dissect the adhesion with an energy L-hook electrode, and not deal with the scar tissue) (n=56). Safety (surgical complications), feasibility (surgical technique replacement rate), and postoperative efficacy (reduction of AFS score, pregnancy, and live birth rate), were each evaluated between groups. RESULTS: No statistically significant differences between the groups were observed in basic preoperative information (P>0.05), while there were significant differences between PG and TG, as well as PG and EG in postoperative AFS scores (PG vs. TG: P=0.007; PG vs. EG: P<0.001) and pregnancy outcome (PG vs. TG: P=0.039; PG vs. EG: P<0.001). No patients had surgical complications such as uterine perforations, moderate or severe fluid overload, heavy uterine bleeding, nor any surgical technique replacements (for example, transfer to use a resectoscope). CONCLUSIONS: Cold scissors ploughing technique in HA is effective, feasible, and safe, and thus worthy of further study.

Growth hormone promotes human endometrial glandular cells proliferation and motion through the GHR-STAT3/5 pathway.

BACKGROUND: This study aims at investigating the effect of growth hormone (GH) on the growth of human endometrial glandular cells (hEGCs) and preliminary exploring its mechanism. METHODS: HEGCs were isolated from the endometrial biopsies and exposed to different dose of GH (0, 50, 100, and 200 ng/mL). Cell proliferation and cell cycle assay, migration assay was performed to investigate the growth and motivation of hEGCs, respectively. Reverse transcription-polymerase chain reaction (RT-PCR), immunocytochemistry (ICC), and western blot (WB) were processed to investigate its related gene or protein expression. RESULTS: The results revealed that GH administration promoted the proliferation, cell cycle, migration, and growth hormone receptors (GHRs) expression of the hEGC. We further inhibited GHRs with AG490, and the inhibitor reversed the effects of GH on cell growth, motion, and the activation of GHR and STAT3/5. CONCLUSIONS: GH promoted hEGCs proliferation and motion, which is GHR-JAK-STAT3/5 signaling pathway-dependent. These findings reveal the essential roles of GH in the hEGCs growth and provide evidence for potential GH therapy in intrauterine adhesion (IUA) treatment.

Logistic regression analyses of factors affecting fertility of intrauterine adhesions patients.

BACKGROUND: Intrauterine adhesion (IUA) prevalence is difficult to measure, but appears to have increased over the last few decades. The reproductive outcomes following hysteroscopic adhesiolysis (HA) for moderate-severe IUAs were unsatisfactory, and few studies have analyzed the clinical characteristics pre-, intra- and post-HA to determine the main risk factors for infertility in patients with IUAs. METHODS: This retrospective observational study included 406 patients, desiring fertility, who had undergone HA between January 1st, 2016 to May 31st, 2017, and had moderate-to-severe IUA [5-12 on the American Fertility Society (AFS) classification scale]. Logistic regression was performed to analyze the data of the clinical characteristics associated with IUA. RESULTS: A total of 406 IUA patients were initially collected. Twenty-six [26] were lost during follow-up or excluded by other criteria; 380 were included in the study with a follow-up period ranging from 2 to 3 years. There were 215 patients (56.6%) that became pregnant, of whom 18 spontaneously miscarried, 5 birthed prematurely (31-36 gestational weeks), 182 delivered at term, and 10 were pregnant at the end of the study. A bivariate and binary logistic regression analysis showed that an age of >30 years, cohesive IUA, lack of increased menstrual volume, and more than 2 times undergoing HA procedure were the risk factors for infertility in IUA patients (P<0.05). CONCLUSIONS: Age, severity of IUA, increased menstrual volume, and HA procedures were the dominant factors affecting reproductive outcomes and may be regarded as potential predictors for evaluating IUA prognosis.

Obstetrical outcome in the third trimester after hysteroscopic adhesiolysis.

BACKGROUND: Intrauterine adhesion (IUA) is caused by adhesion of the uterine cavity due to the damage of endometrium. Hysteroscopic adhesiolysis (HA) is the main treatment. The objective of the study was to investigate the obstetrical outcome in the third trimester of women who previously underwent HA. METHODS: We performed a retrospective cohort study in the university-affiliated hospital. A total of 146 women with a history of HA who had given birth in their third trimester from May 2012 to May 2019 were enrolled (study group), while 292 women with a negative history of HA were matched for maternal age, gravidity, parity, and delivery year. The parameters of obstetrics and infants were investigated to evaluate the change in the third trimester of women with a history of HA. RESULTS: There was no significant difference between study and control groups in gestational weeks, nor in fetal gender distribution, birth weight, Apgar score, fetal distress, and neonatal intensive care unit (NICU) admissions after delivery (P>0.05). In addition, when compared with the control group, women with a history of HA were at a higher risk of placental risks (P<0.05), such as placenta previa (11.6% versus 3.1%), abnormally invasive placenta (AIP) (33.56% versus 2.7%), and retained placenta (42.5% versus 8.6%). This resulted in a significantly higher postpartum hemorrhage (PPH) rate in the study group as compared with that in the control group (8.9% versus 1.0%, P<0.05). Such cases were more likely to be found in patients with severe IUA compared with those who were assessed as mild and moderate. CONCLUSIONS: The history of HA might be an important risk factor inducing placental problems and PPH in the third trimester. More attention should be paid to the labor of pregnant women with a history of HA.

Establishment of an animal model of intrauterine adhesions after surgical abortion and curettage in pregnant rats.

BACKGROUND: Intrauterine adhesion (IUA), also referred to as Asherman syndrome, is a complication with partial or complete adhesion of the uterine cavity due to fibrotic proliferation after endometrial and uterine wall injury of the newly pregnant uterus. Currently, all IUA research models are based on non-pregnant animals which are ill-suited in mimicking its pathogenesis. This study aimed to establish an animal IUA model caused by surgical abortion and curettage in pregnant rats that is more in line with clinical etiology, and to provide a more ideal animal model for further exploration of the pathogenesis and treatment for IUA. METHODS: Fifteen, 13-to-15-day pregnant Sprague-Dawley (SD) rats aged 10 weeks were selected. After incising and removing the embryos, we randomly selected one side of the uterine cavity as the study group (n=15) and scraped it with a curette; the contralateral side of the uterine cavity that underwent no special procedures was used for inclusion in the control group (n=15). Five rats were euthanized and dissected on the 3rd, 7th, and 14th day after surgery respectively. The changes in tissue morphology, the number of endometrial glands, the ratio of endometrial fibrotic area, and the expression level of the transforming growth factor beta (TGF-ß) in the endometrium were compared between the 2 groups. RESULTS: The endometrial granular cell number in the study group on the 7th and 14th day postoperatively was 3.87±0.72 & 2.59±0.90 in each visual field (×100) respectively, which was significantly lower than that of the control group (6.48±0.96 & 7.53±1.10; P<0.05). The ratio of the endometrial stromal fibrotic area in the study group was significantly higher than that of the control group post-operatively on the 3rd day (0.0974±0.0430 vs. 0.0584±0.05110), 7th day (0.1551±0.0348 vs. 0.0328±0.0152), and 14th day (0.1518±0.0477 vs. 0.0311±0.0128) (P<0.05). The endometrial TGF-ß expression in the study group was positive on the 14th day postoperatively while weakly expressed in the control group, and the integrated optical density (IOD) value (5,608.8±2,887.3) was higher than that of the control group (388.3±98.6) (P<0.05). The endometrial TGF-ß expression in the study group and control group was relatively weak on the 3rd and 7th day after surgery, and there was no significant difference when compared with that of the control group (P>0.05). CONCLUSIONS: The establishment of an animal model of IUA after surgical abortion and curettage in pregnant rats was successful and is thus feasible for further studies; the model may be a more effective mimic for the generation of human IUA.