ProblemMaternal-fetal immune tolerance is vital for a healthy pregnancy. However, the metabolic factors governing this balance are not yet clear. This study investigates the role of LDHA in this process and explores how Shoutai Wan (STW) regulates uterine immune cells through LDHA.MethodsA uterine-specific LDHA-knockdown model was established by injecting AAV9-Ldha-RNAi into the uterus of CBA/J mice. Following STW pretreatment, female mice were mated with male BALB/c mice. Gravid uteri were collected on gestational days 6.5, 10.5, and 14.5. Embryo counts were recorded, and tissue morphology was observed using HE staining. Uterine immune cell populations including CD4+ T cells, CD8+ T cells, M1/M2-like macrophages, Th1/Th2 cells, Tregs, and NK cells were analyzed by immunohistochemistry and flow cytometry. IL-4, Foxp3, IFN-γ, and TGF-β1 expression were quantified by qRT-PCR and Western blot.ResultsUterine LDHA knockdown decreased embryo number and disrupted maternal-fetal interface morphology. STW treatment reversed these effects. IHC showed that STW increased CD4+ T cells, F4/80+ macrophages, and NK cells at the maternal-fetal interface of sh-Ldha mice and reduced CD8+ T cell infiltration. Flow cytometry further revealed that STW increased the absolute numbers of uterine Tregs and NK cells in early and mid‑pregnancy, whereas reducing Th1 cell counts at all three time points. Both Western blot and qRT‑PCR analyses confirmed that STW upregulated the protein and mRNA expression of LDHA, IL‑4, TGF‑β1, and Foxp3, while suppressing that of IFN‑γ at the maternal‑fetal interface.ConclusionUterine LDHA inhibition disrupts maternal-fetal immune tolerance in pregnant CBA/J mice. STW treatment restores this balance in sh-Ldha mice, supporting a successful pregnancy.

The decline in oocyte quality is a key factor contributing to the age-related deterioration of female fertility. In women of advanced maternal age, impaired oocyte development is primarily manifested as meiotic abnormalities. An important underlying pathological mechanism is mitochondrial dysfunction, which is often associated with iron overload during aging. Previous studies have suggested that the traditional Chinese medicine formula Bushen Tiaojing Decoction (BSTJD) may improve oocyte quality; however, its specific protective effects in women of advanced maternal age and the related mechanisms remain unclear. This study aimed to investigate whether BSTJD ameliorates meiotic defects in oocytes from aged mice by inhibiting aging-related iron overload. We established both naturally aged and FeSO4-induced iron overload mouse models, followed by BSTJD intervention. In vitro experiments, oocytes from aged mice were cultured and treated with BSTJD-containing serum or adenovirus infection. The results indicated that BSTJD may improve oocyte quality under advanced maternal age conditions by inhibiting iron overload and enhancing mitochondrial function. BSTJD suppressed iron overload-induced reactive oxygen species accumulation, thereby downregulating p53 and p21 expression, promoting CDK1 expression, and reducing ESPL1 expression. Through this regulatory mechanism, BSTJD prevented premature cleavage of chromosomal cohesin by ESPL1, thereby supporting the normal completion of meiosis in aged oocytes and reducing the occurrence of spindle abnormalities and aneuploidy. This study suggests that BSTJD may improve oocyte quality in aged mice by inhibiting iron overload and its downstream p53/CDK1/ESPL1 signaling pathway. This work provides experimental evidence for the clinical application of BSTJD and offers a potential target for intervention strategies aimed at improving oocyte quality decline in women of advanced maternal age.

Background Traditional Chinese medicine (TCM) functions according to the concepts of “holism” and treatment based on syndrome differentiation, and it has achieved good clinical results in treating patients with dysmenorrhea, which is a common gynecological disorder. However, there are currently no international clinical practice guidelines involving TCM therapies for dysmenorrhea. This study aims to establish a protocol for the development of such guidelines. Objective This protocol will provide a road map for the development of the first international clinical practice guidelines on TCM therapy for dysmenorrhea. Methods The guidelines will be developed with reference to the General Rules of Preparation of Diagnosis and Treatment Guideline in Traditional Chinese Medicine and the Western Medicine Guideline and Traditional Chinese Medicine Guideline: Improving Together in Mutual Learning. They will be developed in accordance with the Appraisal of Guidelines for Research and Evaluation II and the World Health Organization guideline handbook and will make recommendations based on systematic reviews. We have established a guideline working group and will formulate clinical questions using the participants, interventions, comparisons, outcomes, and study design framework. The recommendations will be developed through evidence retrieval, synthesis, and the Delphi method. We will consider the preferences and values of patients, as well as the costs and the pros and cons of interventions. Results This work was supported by the National Key R&D Program of China (grant 2019YFC1712000) in 2019. The writing group for guidelines was formed in January 2021. The literature search and screening process began in May 2022. So far, the literature search and 2 rounds of the Delphi method have been completed. The protocol will provide a road map for the development of the first international clinical practice guidelines on TCM therapy for dysmenorrhea. Conclusions The guideline will be developed in accordance with this protocol, which may provide support and evidence for TCM treatment in patients with dysmenorrhea. There is currently a need for clinical practice guidelines in TCM for the treatment of dysmenorrhea. International Registered Report Identifier (IRRID) DERR1-10.2196/77423

BACKGROUND:Endometriosis is an estrogen-dependent, progesterone-resistant gynecological disorder, with dysmenorrhea being the most common manifestation. OBJECTIVE:This study evaluates the efficacy and safety of the Tibetan herbal medication Honghua Ruyi Pill (HHRY) in managing endometriosis-related dysmenorrhea. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS:This is a multi-center, randomized, placebo-controlled, double-blinded clinical trial conducted in seven hospitals in China from July 2021 to January 2023. A total of 164 patients with endometriosis and moderate or severe dysmenorrhea (visual analog scale [VAS] score ≥ 4) were assigned to the treatment or placebo group in a 1:1 ratio by block randomization. Patients received HHRY or placebo twice a day for three consecutive menstrual cycles (MCs) and were followed up for three MCs after stopping the medication. MAIN OUTCOME MEASURES:Primary outcomes were VAS score of the maximum (VASmax) of dysmenorrhea, endometriosis health profile-5 (EHP-5) score, and 5-level EuroQoL 5-dimension version (EQ-5D-5L) score. Secondary outcomes were VASmax of non-menstrual pelvic pain, days of leave taken, emergent use of nonsteroidal anti-inflammatory drugs (NSAIDs) and changes in uterine, cyst and nodule sizes. Safety profiles were assessed based on adverse events, vital signs, serology markers, urinalysis, and liver and kidney function indicators. RESULTS:VASmax of dysmenorrhea, EHP-5 score, EQ-5D-5 L score, and VASmax of non-menstrual pelvic pain were significantly lower in the HHRY group compared to the placebo group at the final follow-up (3.00 vs 5.50, P < 0.001; 4.00 vs 8.00, P < 0.001; 4.00 vs 9.00, P < 0.001; 0.00 vs 1.00, P < 0.001; respectively). The emergent use of NSAIDs for breakthrough pain showed no significant difference between groups (P > 0.999), but the number of patients who had taken time off was significantly different (5.00 vs 14.00, P = 0.028). Sonographic evaluations indicated no significant change in uterine size (P = 0.183) but showed a significant reduction in cyst size (2.09 cm vs 0.20 cm, P = 0.027, sum of 3 diameters of cysts) and nodule size (0.70 cm vs 0.00 cm, P < 0.001, maximum nodule diameter). Safety analysis showed no significant difference in the incidence of adverse events between groups (18.85% vs 28.05%, P = 0.059). CONCLUSION:HHRY can improve dysmenorrhea, chronic pelvic pain, and quality of life in patients with endometriosis. It has a good overall safety profile, and a 3-month treatment can maintain its effects for at least 3 months after the last dose. HHRY may be considered as a new therapeutic option for treating endometriosis-related dysmenorrhea. TRIAL REGISTRATION:Trial registration at ClinicalTrials.gov with registration number: NCT04942015. Please cite this article as: Han M, Liang XF, Gao J, Wang Y, Cao LX, Wang BJ, Wang Y, Zerang Z, Liu JP, Du HL. Honghua Ruyi Pill, a compound herbal medicine, improves endometriosis-related dysmenorrhea: A multicenter randomized, double-blind, placebo-controlled trial. J Integr Med. 2026; 24(2):182-191.

ETHNOPHARMACOLOGICAL RELEVANCE:Recurrent spontaneous abortion (RSA), a common condition, predominantly affects the reproductive system of women of childbearing age. Due to the unique physiological complexity of pregnancy and etiological heterogeneity, RSA is difficult to diagnose and treat clinically. As normal embryonic development depends on specific metabolic processes to maintain an optimal microenvironment, aerobic glycolysis has recently attracted considerable attention as a critical metabolic pathway. The traditional formula Shoutai Wan (STW) has shown obvious clinical effects in the treatment of RSA. However, the underlying molecular mechanisms of STW in managing RSA remain unclear and require further investigation. STUDY OBJECTIVE:To investigate the mechanism by which STW alleviates early pregnancy loss by maintaining glycolytic activity and the acidic microenvironment (AME) in decidual tissues in RSA mice, and by regulating the ERK1/2/HIF-1α signaling pathway to reduce abnormal apoptosis in decidual tissues, restore extracellular matrix (ECM) remodeling, and promote decidual maturation. MATERIALS AND METHODS:Mice were randomly divided into four groups: control, model, dydrogesterone (DYD), and STW. The RSA model was established in all groups except the control. Control and model mice received equivalent volumes of saline via gavage, while DYD (0.2 mL/20 g/day) and STW (0.3 g/20 g/day) groups received corresponding treatments. After 14 consecutive days, females were co-housed with males (2:1 ratio). On gestation day (GD) 6, embryos and the MFI were collected for pathological assessment, including: (1) MFI microenvironment (pH, lactate content); (2) glycolytic metabolism (key enzyme activities/expressions, lactate transporter expression); (3) ERK1/2/HIF-1α signaling pathway (p-ERK1/2, HIF-1α); (4) apoptosis and related factors (Bcl-2/Bax/Caspase-3), apoptosis rates (TUNEL, flow cytometry); (5) ECM remodeling markers (MMP-2, MMP-9, TIMP-1, TIMP-2, Desmin) and decidualization markers (IGFBP-1). The embryo loss rate was calculated on GD14. Additionally, the effects of STW-treated serum on human endometrial stromal cells (hESCs) were examined in vitro to validate the mechanisms. RESULTS:In comparison to the control group, RSA model mice showed a higher level in terms of embryo loss rate, disrupted AME characteristics at the MFI, increased pH, and reduced lactate content. Decreased lactate inhibited activities and expressions of key glycolytic enzymes (HK2, PKM2, LDHA). Downregulation of the ERK1/2/HIF-1α signaling axis resulted in increased apoptosis, impaired ECM remodeling (reduced expressions of MMP-2, MMP-9, TIMP-1/2, and Desmin), and decreased expression of the decidualization marker IGFBP-1. STW and DYD treatments effectively reversed these abnormalities and significantly reduced embryo loss rates. STW demonstrated superior efficacy over DYD, particularly in inhibiting apoptosis and promoting ECM remodeling, confirmed by in vitro experiments. CONCLUSION:Aerobic glycolysis and AME homeostasis at the MFI function as crucial metabolic hubs for maintaining pregnancy. Dysfunction of this system triggers pathological changes, including excessive apoptosis of decidual cells, impaired ECM remodeling, and disrupted decidualization through the ERK1/2/HIF-1α signaling pathway. The core therapeutic mechanism of STW involves effective remodeling of this metabolic-signaling regulatory network, thereby improving early pregnancy outcomes, and promoting embryo implantation.

The incidence of diminished ovarian reserve (DOR) has been increasing annually among younger patients, making it a challenging issue in the field of reproductive health worldwide. Traditional Chinese medicine (TCM) has demonstrated certain therapeutic effects and advantages in addressing the decline of ovarian reserve function, particularly in improving the quantity and quality of oocytes. However, there remains a lack of systematic promotion for the integrated use of TCM and Western medicine in the diagnosis and treatment of DOR. According to the standardized management methods from the China Association of Chinese Medicine, our project team aimed to address the issues of menstrual disorders, sex hormone imbalances, and fertility decline associated with reduced ovarian reserve function. By combining ancient and modern literature with insights from experienced TCM practitioners, we developed an integrated diagnosis and treatment plan. This was achieved through expert interviews and questionnaires, employing methods that integrate TCM with Western approaches, including internal and external treatments as well as long-term adjustments. Our goal is to provide standardized guidance to clinicians for informed decision-making in DOR treatment.

Controlled ovarian hyperstimulation (COH) is a cornerstone of assisted reproductive technologies, yet its effects on endometrial function and embryo implantation remain poorly understood, particularly regarding the role of hypoxia-inducible factor 2α (HIF-2α) signaling. Therefore, the objective of this study was to investigate whether COH-induced endometrial dysfunction impairs mouse embryo implantation through the HIF-2α pathway. A COH mouse model was established using gonadotropin-releasing hormone agonist (GnRH-a)/human menopausal gonadotropin (hMG)/human chorionic gonadotropin (hCG) administration. Embryo implantation status was evaluated on gestational days 5, 6, and 20; endometrial tissues were analyzed for HIF-2α pathway activity via immunohistochemistry (IHC), immunofluorescence (IF), Western blot (WB) and quantitative real-time PCR (qRT-PCR); histological changes were assessed by Jones silver staining and transmission electron microscopy (TEM); serum estradiol (E2), progesterone (P4), and prolactin (PRL) levels were measured by radioimmunoassay (RIA). COH mice exhibited reduced total embryo implantation rates (on day 5, 6 and 20), together with decreased serum E2, P4, and PRL levels. COH mice exhibited preserved luminal epithelium integrity with increased microvillus density and continuous basement membrane structure. qRT-PCR and WB demonstrated significantly downregulated HIF-2α expression at both mRNA and protein levels, accompanied by reduced downstream RAB27B (member of RAS oncogene family)/matrix metalloproteinase 9 (MMP9) and lysyl oxidase (LOX)/adrenomedullin (ADM) signaling, which associated with impaired luminal epithelium detachment and compromised trophoblast invasion. Together, these findings identify HIF-2α as a potential key mediator of COH-induced endometrial microenvironment alterations, revealing molecular mechanisms underlying implantation failure. Importantly, the HIF-2α-RAB27B/MMP9 and HIF-2α-LOX/ADM axes are highlighted as promising therapeutic targets to optimize assisted reproductive outcomes.

Ethnopharmacological relevance The classic TCM prescription, Shoutai Wan (STW), is extensively used in clinical settings to manage threatened miscarriage and Recurrent spontaneous abortion (RSA). The complexity of pregnancy physiology, coupled with diverse etiologies, and the specificity of energy metabolism for normal embryo attachment and development,pose challenges to clinical diagnosis and treatment. The specific molecular mechanisms of how STW regulates these biological processes and contributes to the treatment of RSA remain to be elucidated. Aim of the study This study aims to investigate the causes of early pregnancy loss in RSA mice and explore how STW mitigates this loss. Materials and methods An RSA mouse model will be established and treated with STW and Dydrogesterone (DYD). Embryo loss will be quantified on the 14th day of pregnancy, and embryos will be collected on the 6th and 10th days to observe the embryonic condition and assess pathological changes. The study will analyze aerobic glycolysis and angiogenesis at the maternal-fetal interface (MFI). Additionally, STW on a knockdown LDHA mouse model and Human Endometrial Microvascular Endothelial Cells (HEMECs) in vitro will also be examined to verify the mechanism. Results Compared with the control group, the RSA group exhibited significant embryo loss, and reduced levels of aerobic glycolysis at the MFI, the precarious acidic microenvironment (AME), and the PI3K/AKT/mTOR signaling axis downregulated, leading to impaired angiogenesis, which ameliorated following STW treatment. STW treatment enhanced key aerobic glycolysis enzymes—HK2, PKM2, LDHA—and lactate levels, thereby maintaining the AME and upregulating the PI3K/AKT/mTOR axis. This, in turn, promoted the expression of angiogenesis-related factors (VEGFA and VEGFR2) at the MFI, thereby improving angiogenesis, and the same was seen in sh-LDHA mice. In vitro studies confirmed that STW could counteract the glycolysis decline caused by increased oxygen levels, a recovery that was impaired after LDHA knockdown or PI3K inhibition. Conclusions In RSA mice, disturbances in aerobic glycolysis at the MFI prevent the maintenance of a stable AME, thus impairing angiogenesis and leading to embryo loss, and STW effectively improve early pregnancy outcomes, and laying the foundation for uterine spiral artery remodeling.

Objective In vitro fertilization-embryo transfer (IVF-ET) is a primary treatment method for infertility. It usually requires controlled ovarian hyperstimulation (COH) or repeated COH to obtain more superior oocytes, which could impair ovarian function, reduce embryo quality and limit pregnancy rates. Bushen Tiaojing Fang (BSTJF), our clinically effective prescription for kidney deficiency-related infertility over four decades, while its potential mechanisms to improve pregnancy rates and embryo quality remain unclear.Methods In this study, we focus on the approach of network pharmacology to predict key compounds, core targets, and potential mechanisms of BSTJF for blastocyst induced by repeated COH, mainly including drug-likeness (DL) and oral bioavailability (OB) screening, "herb-component-target-blastocyst" (HCTB) and protein-protein interaction (PPI) networks construction and analysis, GO and KEGG enrichment annotation, molecular docking and basic experiments validation.Results Network pharmacology analysis identified 29 key targets of BSTJF associated with blastocyst development. GO and KEGG enrichment revealed the key targets mainly involved regulating apoptosis to modulate the cell cycle, and demonstrated significant involvement in Cancer, PI3K-Akt, and HIF-1 signaling pathways. In vivo experiments suggested that BSTJF effectively improved the pregnancy rate of repeated COH mice, increased the number of blastocysts while enhancing the rate of high-quality blastocysts. Immunofluorescence (IF) results showed a significant downregulation of P21 and Bax proteins, and a significant upregulation of Cyclin B1 and Bcl-2 proteins in blastocyst of mice treated with BSTJF.Conclusion The study revealed the potential mechanism by which BSTJF could boost embryo quality in mice with repeated COH via activating the PI3K-Akt signaling pathway, which balances the proliferation and apoptosis of blastocyst cells, ultimately restoring normal cell cycle progression and offering new therapeutic strategies for IVF-ET.

Objective: To provide high-quality clinical evidence of the efficacy of Tibetan medicine Honghua Ruyi (HHRY) pills for endometriosis-associated dysmenorrhea. Methods: This study constitutes a multicenter, randomized, double-blind, placebo-controlled trial encompassing a three-menstrual cycle intervention followed by a three-menstrual cycle follow-up period. A total of 164 eligible females with endometriosis-associated dysmenorrhea were randomly divided into HHRY pills and placebo groups in a 1:1 ratio. The primary outcome included dysmenorrhea symptoms assessed using Visual Analog Scale (VAS) scores and quality of life, whereas the secondary outcome measures included the maximum VAS for non-menstrual pelvic pain, duration of pain episodes (in days), frequency and quantity of the consumption of ibuprofen sustained-release capsules (or other non-steroidal anti-inflammatory drugs), and days off work/study for staff/student due to dysmenorrhea, ovarian cyst, and/or pelvic nodule size. The safety was monitored throughout the treatment period. All the analyses were based on the intention-to-treat principle. For continuous outcomes, simple or multiple linear regressions were used to estimate the differences between the HHRY pills and placebo groups, with categorical data expressed as the number and percentage of occurrences. Differences were compared using the chi-square test or Fisher's exact test. The predefined analysis was adjusted for concomitant treatment, a variable considered to be associated with outcomes but unaffected by treatment allocation. Estimates of treatment effects were reported with 95% confidence intervals. Two-tailed P values ≤ .05 were considered statistically significant. Conclusion: Positive results from this trial, upon completion would provide robust evidence for the efficacy and safety of HHRY pills in treating dysmenorrhea in patients with endometriosis.

This study aims to explore the mechanism of Shoutai Pills in treating threatened abortion. According to the random number table method, ICR female mice were randomized into a normal group, a model group, a dydrogesterone group, and a Shoutai Pills group, with 15 mice in each group. Mice were administrated with normal saline(normal and model groups) or the suspension of Shoutai Pills or dydrogesterone by gavage at 9:00 am every day. At 16:00 every day, mice in the normal group were administrated with an equal volume of distilled water, while those in the model, Shoutai Pills, and dydrogesterone groups were administrated with hydrocortisone solution by gavage for 4 consecutive days. ICR female and male mice were caged in a ratio of 2∶1 during the pre-estrous or estrous period. From the first day of pregnancy, drug administration was continued for 5 consecutive days. On day 6, mice were administrated with mifepristone by gavage to establish the model of kidney deficiency-induced abortion. On day 6 of pregnancy, 10 female ICR mice were randomly selected from each group, and the uterus was collected for observation of the pathological changes of trophoblasts at the maternal-fetal interface by hematoxylin-eosin(HE) staining. The protein levels of key enzymes of glycolysis, hexokinase 2(HK2), enolase 1(ENO1), pyruvate kinase M2(PKM2), and lactate dehydrogenase A(LDHA), were determined by Western blot and immunofluorescence. The expression of apoptosis-related proteins including B cell lymphoma-2(Bcl-2), Bcl-2-associated protein X(Bax), and cysteinyl aspartate-specific proteinase-3(caspase-3) was determined by Western blot and real-time PCR. Terminal-deoxynucleoitidyl transferase-mediated nick-end labeling was employed to examine apoptosis. The embryo loss rate of the remaining five female mice was calculated by trypan blue staining method on day 14 of pregnancy. On day 14 of pregnancy, the embryo loss rate of the normal group was 5.00%, which was lower than that(27.78%) in the model group(P&lt;0.05). Dydrogesterone and Shoutai Pills groups showed reduced embryo loss rates(10.26% and 7.50%, respectively) compared with the model group. On day 6 of pregnancy, compared with the normal group, the model group showed down-regulated expression of HK2, ENO1, PKM2, LDHA, and Bcl-2 and up-regulated expression of Bax and caspase-3(P&lt;0.05). Compared with the model group, dydrogesterone and Shoutai Pills up-regulated the expression of HK2, ENO1, PKM2, LDHA, and Bcl-2 and down-regulated the expression of Bax and caspase-3(P&lt;0.05). Compared with that in the normal group, the apoptosis rate in the model group increased(P&lt;0.05). Compared with the model group, dydrogesterone and Shoutai Pills reduced the apoptosis rate(P&lt;0.05). In conclusion, Shoutai Pills can reduce the embryo loss rate and protect embryos by promoting aerobic glycolysis at the maternal-fetal interface and inhibiting the apoptosis of trophoblasts in mice.

Polycystic ovary syndrome (PCOS) is a common disease affecting women of reproductive age; there is a need for interventions to address this condition. Herein, the network pharmacology approach was used to explore the potential of combining Morindae Officinalis Radix (Bajitian) and Radix Salviae (Danshen) for treating PCOS. The bioactive ingredients of the Bajitian-Danshen pair were identified and used for predicting potential therapeutic target genes. Genes related to PCOS were predicted by keyword search from various databases and intersected with the predicted targets of the active components of the Bajitian-Danshen pair to obtain key target genes, which were used for building the “active compound-target gene” pharmacological network. The TCGAbiolinks package in the R environment was used for functional enrichment analysis. In addition, in vivo and in vitro PCOS models were established to explore the effect of the key bioactive compound on the treatment of PCOS and the underlying mechanisms. Histopathological analysis was performed by hematoxylin and eosin staining, while the detection of hormone and cytokine levels was performed by conducting ELISA. Immunofluorescence and western blotting were used for protein expression analysis. Tanshinone IIA (Tan-IIA) was found to be the ingredient with the highest number of target genes. The protein–protein interaction (PPI) network analysis revealed that JUN, FOS, TP53, PTGS2, MMP9, CDKN1A, BCL2, DPP4, and CASP3 were key target genes of Tan-IIA in PCOS. The docking analysis showed the interaction of Tan-IIA with FOS. Thus, the therapeutic potential of Tan-IIA in PCOS and its effect on FOS were evaluated experimentally. A rat model of PCOS was established and subsequently treated with Tan-IIA. Tan-IIA treatment attenuated the deleterious effects associated with PCOS and downregulated TP53, FOS and JUN mRNA and protein expression levels in the ovarian tissue of PCOS rats. In addition, the activation of FOS expression was followed by the exacerbation of the deleterious effect of PCOS and increased expression levels of JUN and TP53. Thus, we concluded that the Bajitian-Danshen pair, especially the Tan-IIA ingredient, counteracts PCOS‑induced damage by possibly regulating the FOS/JUN/TP53 axis.

Ethnopharmacological relevance: Threatened abortion is a common disease among women of childbearing age. Its high incidence rate and unclear etiology, seriously threaten women's physical and mental health. Shoutai Wan (STW) is a traditional Chinese medicine decoction for treating abortion. It has a long history of treating threatened abortion by tonifying the kidney and calming the fetus. However, the mechanism of STW remains unclear. Aim of study: To study the mechanism and potential benefit of STW in pregnant mice with hydrocortisone and mifepristone-induced threatened abortion. Materials and methods: The STW compounds were identified using gas chromatography-mass spectrometry analysis. STW-H, STW-M, or STW-L was separately given 3 mg/ml, 1.5 mg/ml and 0.75 mg/ml STW in the morning, and 2 mg/ml hydrocortisone in the afternoon from gestation day (D) 1-9 and once with 0.4 mg/kg mifepristone on D10. Didroxyprogesterone (0.1 mg/ml) and equal dose pure water were used to replace STW in didroxyprogesterone (DYD) group and model group respectively. The control group used pure water to replace STW, hydrocortisone, and mifepristone. We performed morphological and histological analyses of the maternalfetal interface on day 10. Results: The embryo loss rate in the STW-H and DYD groups was lower than that in the model group. Hematoxylin and eosin (HE) staining suggested that the morphology of maternal-fetal interface was improved in the STW-H and DYD groups. Immunohistochemical (IHC), Quantitative Reverse Transcription Polymerase Chain Reactionstaining (qRT-PCR), and Western blot (WB) results indicated that HIF-1 alpha expression in the maternal-fetal interface of the STW-H and DYD groups was higher than that in model group. The activities of HK, PKM, LDH and the concentration of lactic acid in the STW-H and DYD groups were higher than those in model group. Furthermore, the protein and mRNA levels of HK2, PKM2, LDHA, MCT4, and GPR81 were higher in the STW-H and DYD groups than those in the model group. Conclusions: STW can reduce the pregnancy loss rate by regulating the glycolysis balance at the maternal-fetal interface of kidney deficiency threatened abortion model mice.

The effects of repeated controlled ovarian stimulation (COS) on the female reproductive system are still controversial. This study investigated the effects of repeated COS on the ovaries and uterus of mice and its possible mechanism. Female ICR (Institute of Cancer Research) mice were subjected to the COS using pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) for 1, 3, 5, and 7 cycles. Serum hormone levels, reactive oxidative stress (ROS), 8-hydroxy-2’-deoxyguanosine (8-OHdG), total antioxidant capacity (T-AOC), and superoxide dismutase (SOD) in the mouse ovary and uterus were analyzed by ELISA. The morphology of the ovary and endometrium, ovarian apoptosis, and expressions of the vascular endothelial growth factor (VEGF), leukemia inhibitory factor (LIF), PI3K, AKT, Bax, and Bcl-2 in the ovarian and uterine tissues were tested by hematoxylin–eosin (HE) staining, immunohistochemistry, and western blot. The results showed that repeated COS significantly decreased the hormone level (estradiol, progesterone and anti-Müllerian hormone), high-quality of the MII oocyte ratio, oocyte and embryo number, antioxidant capacity (T-AOC, SOD activity), and the protein level of Bcl-2, LIF, and VEGF, but increased the oxidative damage (ROS, 8-OHdG content), embryo fragment ratio, and expression of pro-apoptotic protein Bax. In addition, the expressions of p-PI3K and p-AKT also decreased with the increase of COS cycle. In conclusion, repeated COS causes ovarian and uterus damage possibly through the PI3K/AKT signaling pathway, and this finding may provide some experimental basis for guiding clinical treatment.

目的 研究补肾助孕方对超促排卵小鼠着床窗口(WOI)子宫内膜胎盘蛋白14(PP14)及子宫自然杀伤(uNK)细胞的影响,探讨其改善胚胎着床的免疫作用机制.方法 将60只动情周期规律的雌性昆明小鼠随机分为正常组、模型组和补肾组.观察小鼠孕第8天胚胎发育和着床情况;HE染色观察小鼠WOI子宫内膜厚度和腺体变化,流式细胞术检测子宫内膜uNK细胞标志物CD49b+及外周血CD4+T、CD8+T细胞水平,放射免疫法检测血清雌二醇(E2)和孕酮(P)水平,免疫组织化学法检测子宫内膜PP14的表达.结果 与正常组比较,模型组小鼠妊娠率和平均胚胎着床数降低(P＜0.05),子宫内膜厚度及腺体数量下降,CD49b+uNK细胞及PP14表达降低,血清E2和P水平及外周血CD8+T细胞水平降低(P＜0.05).与模型组比较,补肾组小鼠妊娠率和平均胚胎着床数增加(P＜0.05);子宫内膜组织结构改善,厚度及腺体数量增加,CD49b+uNK细胞及PP14表达增加,血清E2和P水平及外周血CD8+T细胞水平升高(P＜0.05).各组小鼠阴栓率和外周血CD4+T细胞水平无统计学差异(P＞0.05).结论 补肾助孕方通过调控WOI子宫内膜免疫因素及容受性改善超促排卵小鼠胚胎发育和着床.

复发性流产是生殖医学领域常见的一种妊娠并发症,严重威胁女性身心健康;其病因及发病机制复杂,滋养细胞的凋亡、迁移、侵袭和胎盘血管生成、上皮间充质转化、免疫耐受和子宫内膜蜕膜化等因素参与了复发性流产的过程.磷脂酰肌醇-3 激酶/蛋白激酶B(PI3K-AKT)信号通路参与各种细胞功能调节,在细胞生长、凋亡、血管生成和细胞的迁移、侵袭等方面发挥着重要作用.本文就近年来PI3K-AKT通路参与复发性流产的机制研究进展进行综述,并对今后的研究方向进行探讨及展望.

Morroniside is the main ingredient of Cornus officinalis and has a variety of biological activities including antioxidative effects. Ovarian granulosa cells (GCs) are responsible for regulating the development and atresia of follicles, which are susceptible to oxidative stress. In this study, we determined whether morroniside can inhibit the oxidative stress of GCs induced by hydrogen peroxide (H2O2), leading to improved oocyte quality. The oxidative damage and apoptosis of ovarian GCs cultured in vitro were induced by the addition of H2O2. After pretreatment with morroniside, the levels of ROS, MDA, and 8-OHdG in ovarian GCs were significantly decreased. Morroniside significantly upregulated p-Nrf2 and promoted the nuclear translocation of Nrf2, which transcriptionally activated antioxidant SOD and NQO1. In addition, morroniside significantly regulated the levels of apoptosis-related proteins Bax, Bcl-2, cleaved caspase-9, and cleaved caspase-3 via the p38 and JNK pathways. These results suggest that morroniside can reduce the oxidative damage and apoptosis of ovarian GCs induced by H2O2.

At present, the success rate of embryo implantation is 30%-40%, of which 50%-75% of pregnancy failure originates from abnormal embryo implantation. Embryo implantation is a key link in the establishment of normal pregnancy. The main causes of embryo implantation failure are the damage of endometrial receptivity and the failure of endometrial stromal decidualization. Embryo implantation involves a range of signaling molecules and cytokines, and many studies have shown that connexin proteins are involved in the regulation of embryo implantation and endometrial decidualization. This article reviews the role of connexin in endometrial receptivity, decidualization, vascular remodeling, implantation and its regulatory mechanism, in order to provide theoretical basis for the treatment of infertility and repeated implantation failure patients, and provide new research targets for drug development.

In recent years, the rapid development of assisted reproductive technology has brought good news to infertility patients, but the current success rate of embryo implantation is low. The studies have confirmed that about 50% to 75% of pregnancy failures are due to abnormal embryo implantation. Embryo implantation is a key link in the establishment of a normal pregnancy. Impaired uterine-embryo crosstalk is the main reason for embryo implantation failure. Many studies have confirmed that extracellular vesicles (EVs), as a new method of intercellular communication, play an important role in the process of embryo implantation and can carry drugs as a potential treatment for female infertility. This article reviewed the role of EVs in preimplantation uterine-embryo crosstalk and its regulatory mechanism, in order to provide a theoretical basis for the treatment of infertility and repeated implantation failure patients.