Spontaneous abortion (SA) is a common adverse outcome of early pregnancy, yet its underlying pathophysiological mechanisms remain incompletely understood. Accumulating evidence suggests that dysregulated inflammatory responses at the maternal-fetal interface play a critical role in pregnancy loss. However, the potential associations between alterations in gut microbiota, metabolic disturbances, and localized decidual inflammation in patients with SA have not been systematically characterized. Women with SA (n = 30) and those with normal early pregnancy (NP, n = 28) were enrolled in this study. Proinflammatory cytokines were quantified in decidual tissue homogenates, and histopathological and molecular analyses were performed to evaluate inflammatory activation at the maternal-fetal interface. The gut microbiota composition was profiled using shotgun metagenomic sequencing, while metabolic alterations in the feces were assessed by untargeted metabolomics. Integrated multi-omics analyses were conducted to explore associations among gut microbial dysbiosis, metabolic perturbations, decidual inflammatory signaling, and molecular alterations. Compared with those from the NP group, the decidual tissues from the SA group exhibited significantly elevated levels of IL-1β and TNF-α (1.49-fold and 1.51-fold, both P < 0.0001), accompanied by pronounced histopathological abnormalities. Enhanced activation of the NF-κB signaling pathway was observed at the maternal–fetal interface in SA patients. Metagenomic analyses revealed distinct differences in the gut microbiota composition and community structure between the two groups, with differentially abundant bacterial taxa identified (LDA score > 2.0). Consistent with these findings, fecal metabolomic profiling clearly revealed differences between SA and NP patients, with differentially abundant metabolites (VIP > 1.0, adjusted P < 0.05) predominantly enriched in lipid metabolism, amino acid metabolism, and immune-related pathways. In addition, the expression of leucine-rich repeat-containing G protein-coupled receptor 6 was significantly upregulated (P < 0.0001) in the decidual tissue of SA patients. These findings indicate that SA is associated with localized inflammatory activation at the maternal-fetal interface, dysregulation of decidual molecular activity, gut microbiota dysbiosis, and metabolic perturbations. Integrated multi-omics analyses suggest potential interactions among these factors that may be linked to decidual dysfunction during early pregnancy, providing new insights into the complex pathophysiology of SA.

Oxidative stress occurs in trophoblast cells during the development of tubal pregnancy (TP), compared to normal pregnancy. It has been demonstrated that previous studies have shown that Tanshinone IIA (TSA) can increase reactive oxygen species (ROS) levels and exacerbate oxidative stress in tumor cells, while its effects on trophoblast cells or affects cuproptosis pathways remain unclear. We collected chorionic tissue from patients with normal intrauterine pregnancies (NP) or TP to detect the expression of related proteins. HTR-8/SVneo cells were cultured in vitro and treated with Elesclomol, CuCl2 and/or TSA, tetrathiomolybdate (TTM). The expressions of proteins such as DLAT, DLST, FDX1, Lipo-DLAT, Lipo-DLST, Bax, and Bcl-2 were measured. Mitochondrial membrane potential, cell apoptosis, and cell function were also assessed. The concentration of TSA added to HTR8-SVneo cells was 30 µM. The protein expression of DLAT, DLST, Lipo-DLAT, Lipo-DLST monomers, FDX1 and Bcl-2/ Bax was downregulated by the addition of Elesclomol and CuCl2 intervention in the cells. Meanwhile, the levels of reactive oxygen species (ROS) increased, mitochondrial membrane potential decreased, cell apoptosis increased, and cell invasion and migration were attenuated. The addition of TSA enhanced these effects, while the addition of TTM mitigated them. TSA can promote oxidative stress in HTR-8/SVneo cells, leading to cell apoptosis. This process can be reversed by copper chelator.

Cervical cancer (CC) is a serious risk to women's health; it is necessary to explore less toxic and more effective therapies to cure CC. Triptolide (Tri) is the principal active constituent found in "Tripterygium Wilford," has been shown to have antitumor effects. This study set up to demonstrate whether Tri is capable of inducing ferroptosis in CC cells and its potential mechanism. In vitro, Tri was used to treat CC cells, and lipid peroxidation levels in CC cells were detected by flow cytometry, immunofluorescence, and other experiments; the molecular mechanism of Tri treatment of CC was explored by western blot; moreover, the regulatory effects of Tri on the NRF2/GPX4/xCT axis were verified by overexpressing NRF2 in reverse. In vivo, CC cells tumor-bearing mice were constructed to observe the effect of Tri treatment on tumor growth. In vitro, we have demonstrated that Tri prevents the growth and migration of CC cells. Further investigation revealed that Tri substantially enhances ferroptosis in CC cells by increasing lipid peroxidation accumulation. Mechanically, Tri significantly reduced the expression of NRF2, leading to a corresponding repression of the NRF2 downstream targets GPX4 and xCT. Moreover, overexpressing of NRF2 effectively reversed the impact of Tri on ferroptosis in CC cells. Additionally, animal experiments indicted that Tri markedly inhibited tumor size in nude mice by inhibiting the NRF2/GPX4/xCT axis. Tri exerts antitumor effects by triggering ferroptosis in CC cells through the NRF2/GPX4/xCT axis.

PURPOSE Polycystic ovary syndrome (PCOS) is the most common metabolic and endocrine disorder affecting women of reproductive age. The pathogenesis of PCOS is influenced by factors such as race, genetics, environment, hyperandrogenemia, hyperinsulinemia, and obesity. However, the molecular mechanisms linking RNA modification and PCOS remain underexplored. This study aims to investigate the potential genetic and molecular pathways connecting RNA modification with PCOS through bioinformatics analyses.. METHODS The GSE34526, GSE5850, and GSE98421 datasets were obtained from the National Center for Biotechnology Information Gene Expression Omnibus database. We identified intersecting differentially expressed genes (DEGs) and RNA modification-related genes within the GSE34526 dataset and visualized the overlaps using a Venn diagram. Subsequent analyses included Gene Ontology (GO), pathway enrichment (Kyoto Encyclopedia of Genes and Genomes), gene set enrichment analysis (GSEA), gene set variation analysis (GSVA), and immune infiltration analysis. Additionally, we constructed a protein-protein interaction network as well as mRNA-miRNA, mRNA-RNA binding protein, and mRNA-transcription factor (TF) regulatory networks. The expression and receiver operating characteristic curves of hub genes were also identified. RESULTS The expression of several RNA modification-related DEGs (RMRDEGs) (ALYREF, NUDT1, AGO2, TET2, YTHDF2, and TRMT61B) showed significant differences in PCOS patients. GSEA and GSVA indicated that RMRDEGs were enriched in the hedgehog, MAPK, JAK STAT, and Notch pathways. Key transcription factors, including SP7, KLF8, HCFC1, IRF1, and MLLT1, were identified in the TF regulatory networks. CONCLUSIONS These findings suggest that there are gene and miRNA profile alterations exist in PCOS patients and highlight immune-related differences. This knowledge could pave the way for new research directions in the diagnosis and treatment of PCOS.

ETHNOPHARMACOLOGICAL RELEVANCE:Modified Shoutai Pill, also called Jianwei Shoutai Pill (JSP), is a traditional Chinese medicine prescription that has been used as an effective agent for the treatment of miscarriage.AIM OF THE STUDY:To explore the potential molecular mechanism of JSP against recurrent pregnancy loss (RPL).MATERIALS AND METHODS:In vivo, CBA/J mated DBA/2 mice were used to conduct RPL model, while CBA/J mated BALB/c mice were seen as the control group. Mice were orally administered with JSP, Fer-1 (a ferroptosis inhibitor) or distilled water from day 0.5-12.5 of gestation (GD 0.5-12.5). Pregnancy outcomes were analyzed and ferroptosis related indexes of the whole implantation sites were measured on GD 12.5. In vitro, human trophoblast cell line HTR-8/SVneo was cultured and treated with RAS-selective lethal small molecule 3 (RSL3) (a ferroptosis agonist) or different concentrations of JSP. Then, ferroptosis related indexes were tested to analyze whether JSP could inhibit ferroptosis in HTR-8/SVneo cells.RESULTS:In vivo consequences demonstrated that JSP or Fer-1 alleviated pregnancy outcomes including lower resorption rate and abortion rate. In addition, excessive iron accumulation and MDA level were inhibited, while GSH and GPX content were raised under JSP or Fer-1 exposure. Also, JSP or Fer-1 enhanced protein expressions of GPX4 and SLC7A11 which suppress ferroptosis, and lightened protein expression of ACSL4 which boosts ferroptosis. In vitro, JSP rescued HTR-8/SVneo cell death and migration ability that were injured by RSL3. Furthermore, JSP inhibited RSL3-induced intracellular reactive oxygen species (ROS), lipid ROS and iron deposition.CONCLUSIONS:Collectively, our findings illustrated that the mechanism of JSP in treating RPL might be related to inhibiting ferroptosis, which provided a novel insight into the application of JSP in RPL intervention.

Background With the decline of global fertility, drug therapeutic of ectopic pregnancy is of great significance. Lithospermum erythrorhizon is using for embryo killing as herbal medicine. Shikonin is the critical nucleus of Lithospermum erythrorhizon; however, the mechanism is still unclear. The study aimed to explore the mechanism of shikonin against ectopic pregnancy.Material and Methods In this study, we examined the viability and LDH release of HTR-8/SVneo cells by assays, observed pore formation in cell membranes by microscopy imaging and PI staining, and IL-1β release by WB and ELISA assay kit. Then, we used network pharmacology to analyse the potential interaction between shikonin, ectopic pregnancy and pyroptosis and used molecular docking techniques to verify interactions between shikonin and core common targets. Finally, western blotting and immunofluorescence assay were used to explore the mechanism of shikonin-inducing pyroptosis of HTR-8/SVneo cells.Results Shikonin could cause a significant inhibition of HTR-8/SVneo cell viability in a concentration- and time-dependent manner. In HTR-8/SVneo cells, shikonin-induced cell swelling, bubble formation, an increase in the release of lactate dehydrogenase (LDH) and up-regulation of several pyroptosis-associated factors. And network pharmacology showed that The main targets of shikonin-ectopic pregnancy-pyroptosis were IL-1β and caspase-1, and molecular docking results showed that shikonin can closely bind to IL-1β, caspase-1 and GSDMD. Additionally, the necroptosis inhibitor GSK’872 could not suppress the expression of mature-IL-1β and prevent the pyroptosis phenotype from developing. However, the nucleotide oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inhibitor MCC-950 could downregulate the expression of pyroptosis-associated factors and prevent the pyroptosis phenotype from developing. Shikonin led to an elevation in the expression of cathepsin B (CTSB), and the CTSB inhibitor CA-074 abolished pyroptosis induced by shikonin; however, the NLRP3 inhibitor MCC-950 could not inhibit the expression of CTSB.Conclusions Our results suggest that shikonin activates CTSB to induce NLRP3-dependent pyroptosis in HTR-8/SVneo cells. This study has important clinical implications for the treatment of ectopic pregnancy.

Current studies have indicated that insufficient trophoblast epithelial-mesenchymal transition (EMT), migration and invasion are crucial for spontaneous abortion (SA) occurrence and development. Exosomal miRNAs play significant roles in embryonic development and cellular communication. Hereon, we explored the roles of serum exosomes derived from SA patients on trophoblast EMT, migration and invasion. Exosomes were isolated from normal control (NC) patients with abortion for unplanned pregnancy and SA patients, then characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and western blotting. Exosomal miRNA profiles were identified by miRNA sequencing. The effects of serum exosomes on trophoblast migration and invasion were detected by scratch wound healing and transwell assays, and other potential mechanisms were revealed by quantitative real-time PCR (RT-PCR), western blotting and dual-luciferase reporter assay. Finally, animal experiments were used to explore the effects of exosomal miR-410-3p on embryo absorption in mice. The serum exosomes from SA patients inhibited trophoblast EMT and reduced their migration and invasion ability in vitro. The miRNA sequencing showed that miR-410-3p was upregulated in SA serum exosomes. The functional experiments showed that SA serum exosomes restrained trophoblast EMT, migration and invasion by releasing miR-410-3p. Mechanistically, SA serum exosomal miR-410-3p inhibited trophoblast cell EMT, migration and invasion by targeting TNF receptor-associated factor 6 (TRAF6) at the post-transcriptional level. Besides, SA serum exosomal miR-410-3p inhibited the p38 MAPK signalling pathway by targeting TRAF6 in trophoblasts. Moreover, milk exosomes loaded with miR-410-3p mimic reached the maternal-fetal interface and aggravated embryo absorption in female mice. Clinically, miR-410-3p and TRAF6 expression were abnormal and negatively correlated in the placental villi of SA patients. Our findings indicated that exosome-derived miR-410-3p plays an important role between SA serum and trophoblasts in intercellular communication, suggesting a novel mechanism by which serum exosomal miRNA regulates trophoblasts in SA patients.

Background In this study, we conducted an integrated study of the diagnostic value of MiR-223 in ectopic pregnancy (EP). Methods We used GSE44731 downloaded from GEO and GEO2R to identify differentially expressed miRNA. The hub genes corresponding to the differential miRNA were then identified by using the Xiantao academic tool, GO (Gene Ontology), and KEGG (Kyoto Encyclopedia of Genes and Genomes). Afterward, we used the miEAA database to perform gene set enrichment analysis (GSEA) of differential miRNA, and used Xiantao academic tools again to conduct the ceRNA network based on the target genes. Protein–protein interaction (PPI) network construction and lncRNA of hub miRNA target genes were then predicted by the starbase database. For validation, the villus tissue from intrauterine pregnancy and tubal pregnancy was collected and assayed by qPCR. Results In total 19 differentially expressed miRNAs were screened out, of which MiR-223 had a relatively clear diagnostic significance. Hub genes were enriched and analyzed by GO, KEGG, and GSEA, and the results showed that regulation of NF-κB and other signaling pathways are primarily enriched in ectopic pregnancy. We also obtained 215 key genes from PPI analysis. Our ceRNA analysis indicated that LRRC75A-AS1 and PITPNA-AS1 were associated with MiR-223, and the expression of MiR-223 in qPCR was significantly high in tubal pregnancy group. Conclusion We found that MiR-223 can be used in the diagnosis of EP. Our findings provide valuable information and direction for future research into novel targets for EP diagnosis.

Objective To study the effects of Huayu Xiaozheng granule containing serum on the proliferation,invasion,migration,integrin β3(ITGB3)and fibrinogen alpha chain(FGA)expression of human chorionic trophoblast cells HTR-8/SVneo(HTR-8/SVneo).Methods CCK8 method was used to detect the effect of Huayu Xiaozheng granule containing serum on the proliferation of HTR-8/SVneo cells;transwell and cell scratch were used to detect the effects of Huayu Xiaozheng granule containing serum(low,medium and high concentration groups)on the invasion and migration of HTR-8/SVneo cells;the mRNA expressions of FGA and ITGB3 were detected by qRT-PCR;the expressions of FGA and ITGB3 proteins were detected by Western blot.Results The high,medium and low concentration groups of Huayu Xiaozheng granule containing serum significantly inhibited the activity of HTR-8/SVneo cells.Compared with the control group,the serum containing each dose of Huayu Xiaozheng granule inhibited the proliferation of HTR-8/SVneo cells in varying degrees(P＜0.05);the number of HTR-8/SVneo cells passing through the filtration membrane of Transwell chamber in different doses was lower than that in the blank group,and the membrane penetration rate of HTR-8/SVneo cells in the low dose group was the lowest(P＜0.05);the expression of FGA and ITGB3 protein was down-regulated in the Huayu Xiaozheng granule containing serum group;the expression of FGA and ITGB3 mRNA in the low concentration Huayu Xiaozheng granule containing serum group was down-regulated,and the expression of ITGB3 mRNA in the medium and high concentration Huayu Xiaozheng granule containing serum group was down-regulated.Conclusion Huayu Xiaozheng granule may inhibit HTR-8/SVneo cell adhesion by regulating FGA/ITGB3 pathway,so as to reduce cell activity.

子宫腺肌病常见于育龄期女性,具有侵袭性、激素依赖性,症状多样且进行性加重,病情沉疴缠绵.中医认为其乃离经之血蓄积成癥,治疗多以活血化瘀立法.邓高丕教授结合多年临证经验发现,络病证治体系相较瘀血理论更体现子宫腺肌病的病位、病机、病程等,从络病论治此病,在常规活血化瘀基础上更强调络病成因的辨治和入络药物的使用,临证以通络为法,选用辛味通络药、虫类药、藤类药、利水排脓通络药、补虚养络药等,同时结合患者生理特点、主要诉求个体化施治,以卫护脉络、舒畅脉络,使血不离经,癥块缓消.

目的 通过网络药理学和体外实验探索桂枝茯苓丸作用于卵巢癌的分子机制.方法 从公共数据库中获取桂枝茯苓丸的活性成分以及桂枝茯苓丸和卵巢癌的潜在靶点,然后构建蛋白-蛋白质相互作用(PPI)网络、活性成分-靶点网络及疾病-药物-活性成分-靶点网络,并通过DAVID数据库进行GO和KEGG富集分析,利用AutodockTools软件对核心活性成分和核心靶点进行分子对接.基于网络药理学结果,选择不同浓度桂枝茯苓丸含药血清干预SKOV3细胞,以流式细胞术检测细胞凋亡水平,通过蛋白免疫印迹法对PI3K/Akt通路及凋亡相关蛋白进行验证.结果 筛选得到桂枝茯苓丸476个与活性成分关联的靶点,117个与卵巢癌相关基因,其中有20个共同靶点,这些靶标与细胞凋亡、细胞增殖和分化、蛋白质密切相关,并可能通过癌症中的蛋白聚糖、癌症中的碳代谢、粘着斑、PI3K/Akt信号通路、ErbB信号通路等25个相关途径发挥作用.体外细胞实验证实,桂枝茯苓丸能以浓度和时间依赖性地抑制SKOV3细胞增殖,可以抑制PI3K/Akt通路从而促进SKOV3细胞凋亡.结论 桂枝茯苓丸可通过多成分、多靶点、多通路治疗卵巢癌,初步验证桂枝茯苓丸能抑制PI3K/Akt通路促进细胞凋亡,发挥治疗卵巢癌的作用.

总结邓高丕从固冲通络辨治先兆流产合并宫腔积液经验.邓高丕认为冲任不固为先兆流产合并宫腔积液的核心病机,瘀阻胞络是其重要病理因素.其治疗强调治病与安胎并举,以补肾健脾、活血通络、化瘀止血为主要治法,固冲补肾以平为期,通络祛瘀,中病即止.

Long non-coding RNA (lncRNA) anomalies cause early ovarian failure. LncRNA nuclear enriched abundant transcript 1 (NEAT1) was down-regulated in premature ovarian failure (POF) mice and connected to the illness, however, the mechanism remained unclear. The levels of gene and protein were measured by using quantitative real-time polymerase chain reaction, Western blot, and immunofluorescence. Follicle stimulating hormone (FSH), estradiol (E2), and luteinizing hormone (LH) levels were determined using enzyme-linked immunosorbent assay (ELISA). 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and flow cytometry were used to determine cell viability and apoptosis. The interaction of NEAT1, miR-654, and stanniocalcin-2 (STC2) was verified by dual-luciferase reporter assay or RNA binding protein immunoprecipitation (RIP) assays. The results showed NEAT1 and STC2 down-regulated, while miR-654 up-regulated in POF mice. Overexpression of NEAT1 reduced apoptosis and autophagy in cyclophosphamide (CTX)-treated ovarian granulosa cells (OGCs), and Bax, cleaved-caspase3, LC3B, LC3II/LC3I ratio were decreased and Bcl-2 and p62 were raised. NEAT1 suppressed miR-654 expression by directly targeting miR-654. The inhibition of NEAT1 overexpression on apoptosis and autophagy in OGCs was reversed by miR-654 mimics. STC2 was a target gene of miR-654, and miR-654 inhibitor reduced the apoptosis and autophagy by regulating the STC2/MAPK axis. To sum up, NEAT1 reduced miR-654 expression and modulated the STC2/MAPK pathway to decrease apoptosis and autophagy in POF, indicating a potential therapeutic target.

目的 探讨大补元煎含药血清经FOXO3a介导的磷脂酰肌醇激酶(PI3K)/蛋白激酶B(AKT)信号通路对卵巢储备功能减退改善的作用及机制.方法 取36只健康SD大鼠随机分为空白组(6只)、模型组(6只)、大补元煎高剂量组(6只)、大补元煎中剂量组(6只)、大补元煎低剂量组(6只)和西药组(6只),均注射环磷酰胺75 mg/kg造模,在进行相应的治疗后,比较各组大鼠的卵巢储备功能.结果 HE染色结果显示,与模型组比较,大补元煎高、中、低剂量组和西药组大鼠卵巢组织的卵泡数量、颗粒层厚度、层次数,排列密集程度明显增加,闭锁卵泡数量减少.空白组大鼠的血清雌二醇(E2)、抗米勒管激素(AMH)、抑制素B(INH-B)高于其他各组大鼠,促卵泡刺激素(FSH)、促黄体生成素(LH)低于其他各组大鼠,差异有统计学意义(P<0.05);大补元煎高、中、低剂量组及西药组大鼠的血清E2、AMH、INH-B高于模型组,FSH、LH低于模型组,差异有统计学意义(P<0.05);大补元煎高剂量组大鼠的血清E2、AMH、INH-B高于大补元煎中、低剂量组及西药组,FSH、LH低于大补元煎中、低剂量组及西药组,差异有统计学意义(P<0.05);大补元煎中、低剂量组及西药组大鼠的血清E2、AMH、INH-B、FSH、LH比较,差异无统计学意义(P>0.05).空白组、大补元煎高剂量组大鼠的FOXO3a表达量低于其他各组大鼠,PI3K、AKT表达量高于其他各组大鼠,差异有统计学意义(P<0.05);空白组、大补元煎高剂量组大鼠的FOXO3a、PI3K、AKT表达量比较,差异无统计学意义(P>0.05);大补元煎高、中、低剂量组及西药组大鼠的FOXO3a表达量低于模型组,差异有统计学意义(P<0.05);大补元煎中、低剂量组及西药组大鼠的FOXO3a、PI3K、AKT表达量比较,差异无统计学意义(P>0.05).结论 大补元煎可固本培元、大补气血,在调理卵巢功能下降方面有显著疗效,有缓解大鼠卵巢早衰症状的作用,此作用与大补元煎调控体内FOXO3a、PI3K和AKT表达量机制密切相关.

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.

目的:基于黏附斑激酶(FAK)为中心的细胞外基质-整合素-细胞骨架跨膜信号系统,探讨化瘀消癥复方对输卵管妊娠滋养细胞迁移的调控机制.方法:对输卵管妊娠绒毛滋养层细胞进行体外培养与鉴定,以含化瘀消癥复方水提液的培养基对输卵管妊娠滋养细胞进行培养,分为A(空白)组、B(FAK抑制剂Y15)组、C(Y15+中药中剂量)组、D(中药低剂量)组、E(中药中剂量)组、F(中药高剂量)组;应用Transwell法检测各组药物对滋养细胞迁移力的影响,Western Blot法检测各组药物对FAK、p-FAK、整合素β 1(Integrin β1)、桩蛋白(Paxillin)、尿激酶型纤溶酶激活物(uPA)蛋白表达的影响.结果:与A组比较,各给药组均能显著抑制细胞的迁移能力(P＜0.05),各中药组呈一定的剂量依赖性;与A组、B组比较,C组、F组均可下调FAK、p-FAK、Integrin β 1、Paxillin、uPA蛋白表达水平(P＜0.05);各中药组对蛋白的影响呈一定的浓度依赖性.结论:化瘀消癥复方可能发挥与FAK抑制剂类似的效应,起到抑制输卵管妊娠滋养细胞迁移的作用,其机制可能是与下调Integrin β1,抑制FAK的信号转导,引起下游Paxillin的表达减少,负调控uPA,进而降调滋养细胞黏附斑激酶跨膜信号系统活性相关.

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.

Ovarian cancer (OV) is a complex gynecological disease, and its molecular characteristics are not clear. In this study, the molecular characteristics of OV subtypes based on metabolic genes were explored through the comprehensive analysis of genomic data. A set of transcriptome data of 2752 known metabolic genes was used as a seed for performing non negative matrix factorization (NMF) clustering. Three subtypes of OV (C1, C2 and C3) were found in analysis. The proportion of various immune cells in C1 was higher than that in C2 and C3 subtypes. The expression level of immune checkpoint genes TNFRSF9 in C1 was higher than that of other subtypes. The activation scores of cell cycle, RTK-RAS, Wnt and angiogenesis pathway and ESTIMATE immune scores in C1 group were higher than those in C2 and C3 groups. In the validation set, grade was significantly correlated with OV subtype C1. Functional analysis showed that the extracellular matrix related items in C1 subtype were significantly different from other subtypes. Drug sensitivity analysis showed that C2 subtype was more sensitive to immunotherapy. Survival analysis of differential genes showed that the expression of PXDN and CXCL11 was significantly correlated with survival. The results of tissue microarray immunohistochemistry showed that the expression of PXDN was significantly correlated with tumor size and pathological grade. Based on the genomics of metabolic genes, a new OV typing method was developed, which improved our understanding of the molecular characteristics of human OV.

目的 研究紫草素对HTR-8/Svneo细胞线粒体自噬和线粒体功能的影响.方法 将人绒毛膜滋养层细胞分为4组:对照组、Mdivi-1组、紫草素组和紫草素+Mdivi-1组.对照组正常培养;Mdivi-1组以5μmol·L-1 Mdivi-1处理;紫草素组用0.8μmol·L-1紫草素干预;紫草素+Mdivi-1组以0.8μmol·L-1紫草素+5μmol·L-1 Mdivi-1干预处理.用流式细胞术检测细胞凋亡率;用ATP检测试剂盒测定细胞ATP含量;用Dihydroethidium(DHE)荧光检测探针测定细胞内超氧化物水平;用蛋白质印迹法测定线粒体自噬相关蛋白表达水平.结果 细胞分组处理24 h后,对照组、Mdivi-1组、紫草素组、紫草素+Mdivi-1组的凋亡率分别为(6.86±0.37)％,(7.14±0.23)％,(39.97±1.52)％ 和(14.63±1.19)％;这4组的ATP含量为100％,(89.13±6.23)％,(52.03±9.12)％和(67.84±10.03)％;这4组的DHE探针荧光强度为23.02±1.56,23.42±0.51,43.96±0.62和36.23±1.89;这4组的PTEN介导的假定激酶蛋白1(PINK1)蛋白表达水平为0.37±0.14,0.57±0.13,1.26±0.15和0.97±0.07.上述指标,紫草素组与对照组比较,差异均有统计学意义(均P<0.01).结论 紫草素能抑制HTR-8/Svneo细胞增殖,通过PINK1/Parkin通路诱导线粒体自噬的发生且伴有线粒体功能损伤,最终发生细胞凋亡.

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.