BackgroundKidney deficiency syndrome (KDS) is the predominant syndrome associated with gynecological reproductive system diseases in traditional Chinese medicine (TCM). However, the diagnostic method is influenced by the subjective experience of doctors, which leads to the ambiguity in differentiation of KDS and poor effect for corresponding treatment.ObjectiveTo explore an objective syndrome differentiation method for KDS in females of reproductive age through machine learning technique.MethodsWe proposed a new deep learning method for the objective differentiation of KDS in females of reproductive age. First, we simultaneously acquired 376 pairs of tongue and facial images. We divided them into a Kidney deficiency syndrome (KDS, n = 182) group and a Non-Kidney deficiency syndrome (NKDS, n = 194) group. Then, we employed two parallel DenseNet structures to extract deep features from tongue and facial images. We further used a deep supervised network strategy to better stabilize the fusion of the two deep features. We used 5-fold cross-validation to evaluate the performance by six indicators: accuracy, precision, recall, F1 score, receiver operating characteristic (ROC) and area under the curve (AUC). Finally, external validation was conducted on an independent test set consisting of 130 patients with a 1:1 ratio of KDS to NKDS cases.ResultsThe model based on tongue images, facial images, and the tongue-face fusion model achieved AUCs of 71.45% ± 6.39%, 89.60% ± 3.33%, and 92.08% ± 4.51%, respectively, with the highest value observed in the fusion model. In external validation, the tongue-face fusion model attained an AUC of 83.53%.ConclusionThe deep learning network model with tongue-face fusion can effectively differentiate KDS.

BACKGROUND:Miscarriage has the characteristics of recurrent attacks and complex etiology, so it is gradually attracted the wide attention of scholars in the fields of reproduction. Potential association between gut microbiome (GM) and pregnancy disorders has been investigated. Jianwei Shoutai pills (JWP), as a representative formula, have been proven to have protective effect in both clinical and experimental research in miscarriage. However, the specific mechanism of JWP in miscarriage through GM remains unclear. PURPOSE:To investigate the underlying mechanism of JWP against miscarriage through the gut-uterus axis. METHODS:The effects of JWP on an RU486-induced rat model of miscarriage were evaluated by embryo resorption rate, vaginal bleeding rate, and appearance of the uterus and embryo. We used 16S rRNA sequencing to measure the extent of the effect of JWP on GM of rats with miscarriage. Bile acid (BA) content of the feces of rats treated with JWP was evaluated by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS). The activation of bile acid-associated receptor, Farnesoid X receptor (FXR), was evaluated by immunofluorescence. The expression level of NLRP3 inflammasome-associated protein was detected by Western blot or Elisa. Fecal microbiota transplantation (FMT) was used to confirm that GM was essential for the therapeutic effect of JWP in miscarriage. RESULTS:JWP significantly ameliorated miscarriage symptoms and embryo resorption rate caused by RU486-induced miscarriage as well as restored the abnormal activation of NLRP3-inflammasome at the maternal-fetal interface. Furthermore, JWP can significantly regulated GM dysbiosis and closely associated with BA metabolism by KEGG pathway prediction analysis. Several BA content were significantly restored by HPLC-MS. The expression of NLRP3 inflammasome-associated protein at maternal-fetal interface was reversed by JWP. Combined with FMT, JWP could regulate activation of NLRP3 at the maternal-fetal interface by BAs produced by GM. CONCLUSION:JWP restored abnormal activation of the NLRP3-inflammasome in an RU486-induced miscarriage rat model, and corrected the BA disorder by regulating imbalance of the GM.

目的 探索阴道菌群近10年来的研究现状并预测其发展趋势.方法 在Web of Science数据库中检索近10年阴道菌群相关的文献,运用可视化软件VOSviewer进行共词分析、文献耦合、合著分析等.结果 阴道菌群领域发文量呈整体上升趋势,目前的主要热点研究分为阴道菌群的多样性、传染病学、细胞生物和基因学、阴道菌群与生殖的关联四大方面.结论 文献可视化研究展现了阴道菌群的研究现状和发展趋势,为相关研究人员提供了借鉴.

Spontaneous abortion (SA) is the most common adverse pregnancy outcome in the first trimester. The causal drivers of SA have become a substantial challenge to reveal and overcome.

The early diagnosis and treatment of ectopic pregnancy (EP) remains a major challenge. Despite a known link between vaginal microbiota and female reproductive health, few studies have focused on the association between vaginal microbiota and pregnancy location. This nested case-control study aimed to characterize the vaginal microbiota in tubal pregnancy (TP) among symptomatic women in early pregnancy. Women with symptomatic early pregnancy of unknown location (PUL) were included in this study. 16S rDNA gene sequencing was performed to assess vaginal microbial diversity and relative abundance. Machine learning and multivariate logistic regression were also used to evaluate the association between Gardnerella and TP. The results indicate that the vaginal microbiome in TP was more diverse (Shannon, p < 0.05) and was different in composition to that of women with intrauterine pregnancy (IUP) (weighted Unifrac, R = 0.08, p = 0.01). The genus Gardnerella was significantly enriched in TP. The XGBoost analysis was able to classify Gardnerella-induced TP more reliably (AUC = 0.621). Moreover, after adjusting potential confounders, our results indicate a robust association between Gardnerella and TP (as a continuous variable, adjusted OR: 12.0, 95% CI: 2.1–67.4, p < 0.01; as a categorical variable (≥0.85%), and adjusted OR: 4.2, 95% CI: 2.0–8.8, p < 0.01). In conclusion, we found that higher virginal Gardnerella levels were associated with TP in women with symptomatic early pregnancy.

Introduction Studies have suggested that the vaginal microbiome and gut microbiome are involved in pregnancy-related diseases, but little exploration of the link with early miscarriage or threatened miscarriage (TM) has been done. Whether the characteristics of the vaginal microbiome and gut microbiome in early pregnancy are related to TM and early pregnancy outcomes remains unclear.Methods and analysis The Microbiome Characteristics in Early Threatened Miscarriage Study (MCETMS) is a prospective investigation that will recruit 326 pregnant women with early TM. Pregnant women will be enrolled at 4–8 weeks of gestation, and their vaginal secretions, faecal samples, clinical data and sociodemographic characteristics will be collected prospectively. Pregnant women with TM will be followed up to 12 weeks of gestation to determine the early pregnancy outcomes (ongoing pregnancy or pregnancy loss). DNA will be extracted from the collected samples and will be analysed by 16S rRNA gene sequencing.Ethics and dissemination The MCETMS study protocol has been approved by the Ethics Committee of the First Affiliated Hospital of Guangzhou Traditional Chinese Medical University (ZYYECK[2020]051). Dissemination of study findings will occur through peer-reviewed journals, conferences and presentations.Trial registration number ChiCTR2000041172.

Background: The relationship between serum folate status and cognitive functions is still controversial. Objective: To evaluate the association between serum tetrahydrofolate and cognitive functions. Methods: A total of 3,132 participants (60–80 years old) from the 2011–2014 NHANES were included in this cross-sectional study. The primary outcome measure was cognitive function assessment, determined by the Consortium to Establish a Registry for Alzheimer’s Disease Word Learning Test (CERAD-WL), CERAD-Delayed Recall Test (CERAD-DR), Animal Fluency Test (AF), Digit Symbol Substitution Test (DSST), and global cognitive score. Generalized linear model (GLM), multivariate logistic regression models, weighted generalized additive models (GAM), and subgroup analyses were performed to evaluate the association between serum tetrahydrofolate and low cognitive functions. Results: In GLM, and the crude model, model 1, model 2 of multivariate logistic regression models, increased serum tetrahydrofolate was associated with reduced cognitive functions via AF, DSST, CERAD-WL, CERAD-DR, and global cognitive score ( p < 0.05). In GAM, the inflection points were 1.1, 2.8, and 2.8 nmol/L tetrahydrofolate, determined by a two-piece wise linear regression model of AF, DSST, and global cognitive score, respectively. Also, in GAM, there were no non-linear relationship between serum tetrahydrofolate and low cognitive functions, as determined by CERAD-WL or CERAD-DR. The results of subgroup analyses found that serum tetrahydrofolate levels and reduced cognitive functions as determined by AF had significant interactions for age and body mass index. The association between high serum tetrahydrofolate level and reduced cognitive functions as determined using DSST, CERAD-WL, CERAD-DR, or global cognitive score had no interaction with the associations between cognition and gender, or age, or so on. Conclusion: High serum tetrahydrofolate level is associated with significantly reduced cognitive function.

The impact of dietary fiber on human papillomavirus (HPV) infection is still underway. The aim of our study was to investigate the association between intake of dietary fiber and HPV infection. Overall, 14,151 eligible women, aged 18-59 years old, who submitted an adequate sample for HPV test, were collected from an ongoing, large scale population-based survey for seven cycles. The association of dietary fiber intake and HPV infection was assessed in multivariate logistic models. For sensitivity analysis, generalized additive model (GAM) and smooth curve fitting were employed to verify the robustness of the results. Among 14,151 eligible participants, intake of dietary fiber was negatively associated with HPV infection. Each additional increase in log(10) dietary fiber consumption was associated with a 57% lower risk of HPV infection (OR, 0.43; 95% CI 0.38-0.48). The result is stable in minimally and fully adjusted model. The possibility of nonlinear association of dietary fiber and HPV infection has been excluded by GAM and smooth curve fitting. There was an inverse linear correlation between intake of dietary fiber and HPV infection. Our findings obtained from NHANES dataset suggested that increasing dietary fiber consumption may be associated with the prevalence of HPV infection.

The features of the vaginal microbiota (VM) community can reflect health status, and they could become new biomarkers for disease diagnosis. During pregnancy, domination of bacteria of the genus Lactobacillus in the VM community is regarded as a keystone because they stabilize the VM by producing antimicrobial compounds and competing adhesion. An altered VM composition provides a marker for adverse pregnancy outcomes. This nested case–control study aimed to characterize the VM in women with a tubal pregnancy (TP) presenting with pain and/or uterine bleeding in early pregnancy. Chinese women with a symptomatic early pregnancy of unknown location were the study cohort. 16S rDNA gene-sequencing of V3–V4 variable regions was done to assess the diversity, structures, taxonomic biomarkers, and classification of the VM community. The primary outcome was the location of the early pregnancy. The VM community in women with a TP showed higher diversity (PD-whole-tree, median: 8.26 vs. 7.08, P = 0.047; Shannon Diversity Index, median: 1.43 vs 0.99, P = 0.03) and showed different structures to those in women with an intrauterine pregnancy (IUP) (R = 0.23, P < 0.01). Bacteria of the genus Lactobacillus were significantly enriched in the IUP group, whereas bacteria of the genera Gardnerella and Prevotella were significantly enriched in the TP group. Lactobacillus abundance could be used to classify the pregnancy location (AUC = 0.81). Non-Lactobacillus-dominated microbiota (≤ 0.85% Lactobacillus) was significantly associated with a TP (adjusted odds ratio: 4.42, 95% confidence interval: 1.33 to 14.71, P = 0.02). In conclusion, among women with a symptomatic early pregnancy, a higher diversity and lower abundance of Lactobacillus in the VM is associated with a TP.

Background: Pregnancy of unknown location (PUL) is a kind of pregnancy that has a positive beta-human chorionic gonadotropin (β-hCG) test result but the location of pregnancy cannot be determined by ultrasound. Early determination of the location of the pregnancy is important for subsequent treatment. However, there is no study on the characteristics of vaginal microbiota in pregnant women with unknown locations and a model for judging it by vaginal microbiota combined with clinical indicators. Therefore, we designed this study to compare the characteristics of vaginal microbiota in intrauterine and ectopic pregnancy populations during pregnancy with unknown locations and to establish a prediction model for pregnancy locations in PUL populations with clinical indicators.Methods: This is a prospective, multicenter cohort study. 576 eligible participants will be included in this study. Vaginal microbiota was collected from all participants at inclusion, and color Doppler ultrasound was performed weekly. After the locations of pregnancy were determined, participants of intrauterine pregnancy were followed up to their early pregnancy outcome, and participants of ectopic pregnancy were followed up until a none-pregnancy level of β-hCG was confirmed. Discussion: The regular method of judging the location of pregnancy is by color Doppler ultrasound and β-hCG test. We hope to provide earlier clinical methods of prediction for women with unknown locations of pregnancy through this study.Trial registration: Chinese Clinical Trial Registry: ChiCTR2000035378, registration date: 9 August 2020. http://www.chictr.org.cn/index.aspx

Purpose: The aim of this study was to evaluate white blood cell (WBC) count as a risk factor related to methotrexate (MTX) treatment failure in patients with ectopic pregnancy (EP).Methods: A total of 236 women diagnosed with EP and treated with a single dose of MTX were included. The exposure variable was WBC count at baseline, and the outcome was MTX treatment outcome. Both a multivariate binary logistics regression model and subgroup analysis were performed to evaluate the association between WBC and MTX non-response.Results: WBC count was associated with the risk of treatment failure, and the odds ratio (OR) in different multivariate models was stable [minimally adjusted model: OR 1.2, 95% confidence interval (CI): 1.0–1.3, p = 0.008; fully adjusted model: OR 1.2, 95% CI: 1.0–1.4, p = 0.026]. For WBCs in group T3 (>8.9 × 109/L), the association between WBC count and treatment failure was significant (minimally adjusted model: OR: 2.0, 95% CI: 1.0–3.8, p = 0.050; fully adjusted model: OR: 2.2, 95% CI: 1.1–5.6, p = 0.034). Subgroup analysis showed that in participants with regular menstruation (OR 1.1, 95% CI: 1.0–1.3), WBC count was significantly different from irregular menstruation (OR 1.8, 95% CI: 1.2–2.8); p for interaction was 0.031.Conclusions: We found a reliable and non-linear relationship between WBC count and MTX treatment failure for EP.

Objective: Genistein obviously inhibits the migration and invasion of various tumor cells. However, its effects on cervical cancer cells have seldom been referred. We aimed to evaluate the effects of genistein on the proliferation, migration and invasion of cervical cancer HeLa cells, the expressions and phosphorylations of proteins related with FAK-paxillin and MAPKs signaling pathways, as well as the expressions of related key genes. Materials and methods: HeLa cells were stimulated with genistein for 24 h and 48 h respectively. After adherence for 2 h, 0 mu M, 12.5 mu M, 25 mu M, 50 mu M and 100 mu M genistein solutions were added in DMEM. Cell proliferation was tested by the CCK-8 assay. After treatment with 100 mu M genistein, the migration ability was detected by the scratch assay. Transwell assay was used to detect cell migration and invasion abilities. Western blot and qRT-PCR were used to detect the expressions of proteins and mRNAs related with FAK-paxillin and MAPKs signaling pathways respectively. Results: The effect of genistein on the proliferation of HeLa cells was proportional to treatment time and drug dose, and the proliferation was inhibited after 24 h and 48 h at 100 mu M. After treatment with 100 mu M genistein, the scratch migration rate was significantly lower than that of the control group at 24 h and 48 h (P < 0.05). Genistein also inhibited the invasion of tumor cells through the upper chamber and Matrigel. The number of invasive cells was significantly lower than that of the control group (P < 0.05). Genistein significantly inhibited the phosphorylations of FAK, paxillin, p38 and p42/44. Compared to the control group, 100 mu M genistein significantly suppressed the mRNA expressions of FAK, paxillin, Snail and twist. Conclusion: Genistein inhibited the migration and invasion of cervical cancer HeLa cells by regulating FAK-paxillin and MAPK signaling pathways in dose-dependent manners. (C) 2020 Taiwan Association of Obstetrics & Gynecology. Publishing services by Elsevier B.V.

Background: In China, approximately 15% of tubal pregnancy patients treated with MTX eventually required surgery because the ectopic mass was ruptured; therefore, it is essential to develop a model to predict the risk of failure with methotrexate treatment in tubal pregnancy. Material/Methods: In this research, 168 patients met the eligibility criteria, and 29 candidate risk factors for treatment failure were collected. Multivariable logistic regression analysis was used to analyze the factors, and a full model was developed. We used a multiple fractional polynomial model and a stepwise model to increase the reliability. Bootstrap resampling for 500 times was used to internally test the prediction model. The integral performance of the model depends on the evaluation of the nomogram, the discriminative performance by receiver operating characteristic (ROC) curve analysis, and calibration. Results: The model showed excellent discrimination and calibration. The area under the ROC curve for the prediction model, mfp model, and stepwise model were 0.879 (95% CI: 0.812-0.942), 0.872 (95% CI: 0.805-0.931), and 0.880 (95% CI: 0.817-0.949), respectively. At a cutoff value of >= 0.40, sensitivity was 60%, specificity was 91%, positive predictive value (PPV) was 81%, and negative predictive value (N PV) was 77%. The model provides a net benefit when clinical decision thresholds are between 0% and 40% of predicted risk. Conclusion: This model indicated good accuracy in predicting methotrexate treatment failure for tubal pregnancy patients.

Unruptured ectopic pregnancy (UEP) is a common cause of morbidity and, occasionally, of mortality in women of reproductive age. Pharmacological intervention is a common therapeutic approach for early-stage UEP. Herein, we investigated the cytotoxic effect and novel mechanism of shikonin, a natural naphthoquinone pigment purified from Lithospermum erythrorhizon, in human trophoblast cells. These data demonstrated that shikonin suppressed proliferation and induced apoptosis in a time-dependent manner in HTR-8/SVneo cells. Shikonin blocked autophagic flux and promoted p62 interaction with caspase 8, resulting in caspase 8 activation. Moreover, shikonin suppressed GLI1 expression, and GLI1 overexpression attenuated shikonin-induced cell apoptosis. Although silencing GLI1 slightly promoted cell apoptosis, p62 overexpression enhanced GLI1 silencing-induced cell apoptosis by activating caspase 8. Furthermore, rapamycin increased shikonin-induced cell apoptosis in HTR-8/SVneo cells, whereas 3-MA attenuated the cytotoxic effect of shikonin. In conclusion, shikonin suppressed trophoblast cell growth by silencing GLI1 and increasing p62 co-mediated activation of caspase 8, which suggested a potential novel therapeutic target for UEP.

Design From July 2016 to June 2018, 36 women with symptomatic early pregnancy around 4-8 weeks of gestation were recruited into the study. Among them, there were 16 women with viable intrauterine pregnancy (VIP), 9 women with spontaneous abortion (SA), and 11 women of EP. Serum exosomal miRNAs were extracted and measured at the first prenatal visit. Statistical analysis was performed to determine the clinical utility of these biomarkers as single markers and as multimarker panels for EP. Results Concentrations of miR-378d in serum exosomes were significantly higher in EP than in VIP and also SA group. As a single marker, miR-378d had the highest specificity of 64% at the sensitivity of 89.1%. Comparatively, both combined panels of hCG, progesterone, miR-100-5p and hCG, progesterone, and miR-215-5P yielded the specificity of 96%. Panels for all markers achieved the highest specificity of 80% at the sensitivity of 91%. Conclusions Although further validation in large-scale prospective studies is necessary, our results suggest that serum exosomal miR-378d, miR-100-5p, and miR-215-5P are promising biomarkers for early EP.

Mercury is a ubiquitous toxic heavy metal associated with an increased risk of female infertility; however, the evidence supporting this is limited and controversial. We aimed to explore the relationship between the total blood mercury and infertility in 1796 selected participants from the National Health and Nutrition Examination Survey (NHANES) (2013–2016). We found no significant association between mercury and infertility based on a fully-adjusted model (OR 1.04; 95 % CI 0.91, 1.19), and the results remained robust in a series of sensitive analysis. However, a non-linear relationship was detected. By a two-piecewise linear regression model and recursive algorithm, we identified an inflection point of 5.278 μg/L, when blood mercury was >5.278 μg/L, a 1-unit increase in mercury (log2) was associated with 157 % greater adjusted odds of infertility (OR 2.57; 95 % CI 1.12, 5.87). Our findings provide new insights to advance the research of the link between mercury and infertility.

Sambucus williamsii Hance has been used in fractures for thousands of years, but research on its active components, such as morroniside, until now had not been carried out. In this study, morroniside was taken as the leading compound, and fourteen derivatives were synthesized. The promotion of osteoblast proliferation effect of the derivatives was evaluated on MC3T3-E1 cells. Five derivatives (2, 3, 4, 5, and 14) showed a good proliferation effect on MC3T3-E1 cells, and their promoted expression effects on OC (Osteocalcin) and ALP (Alkaline phosphatase) in MC3T3-E1 cells were measured. Compound 3 was shown to have the strongest proliferation effect (EC50 = 14.78 ± 1.17 μg/mL) and to significantly promote the expression of OC and ALP.