INTRODUCTION:This study aims to assess the therapeutic mechanisms of Zishen Yutai Pills (ZYP) in treating recurrent spontaneous abortion (RSA) using network pharmacology analysis and in vitro validation of the molecular pathways relating to angiogenesis. METHODS:An online database screening-based strategy was applied for chemical constituent and target screening-based strategy was applied for chemical constituent and target collection, as well as target prediction. Network pharmacological analysis was conducted using protein-protein interaction data. Molecular docking was further employed to validate the active components of ZYP and the central target proteins. The tube formation assay was conducted in human umbilical vein endothelial cells (HUVECs) with the intervention of active constituents in ZYP. The expression of vascular endothelial growth factor A (VEGFA) and protein kinase B (AKT1) in HUVECs was also quantified by Western Blot (WB). RESULTS:Network pharmacological analysis revealed 283 target genes by ZYP in the treatment of RSA. Through protein-protein interaction (PPI) and topological analyses, a total of 57 key targets were identified as exerting central roles. Molecular docking proved that quercetin, β-carotene, resveratrol, and atractylenolide I had strong binding to AKT1 and VEGFA. Elevated levels of AKT1 and VEGFA protein expression were observed in WB analysis following intervention with the active ingredient. DISCUSSION:The findings suggest that ZYP may ameliorate RSA by promoting angiogenesis at the maternal-fetal interface. Key active compounds were predicted to act via the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway, as demonstrated by molecular docking and cell-based assays. CONCLUSION:This study illuminates the comprehensive therapeutic effects of ZYP in RSA treatment, providing a foundation for its further exploration.

BACKGROUND:Atopic dermatitis (AD) is a chronic inflammatory skin disease often exacerbated by psychological stress, yet the underlying mechanisms remain unclear. Wushe Zhiyang Pills (WZP), an oral Chinese patent medicine, is clinically used for dermatological conditions, but its efficacy and mechanisms in stress-aggravated AD have not been defined. PURPOSE:This study aims to evaluate the protective effects of WZP against AD under stress conditions, elucidate its molecular mechanisms, and identify its bioactive compounds. METHODS:A murine AD model was established using repeated DNCB application, with or without chronic restraint stress to simulate psychological stress exposure. AD-like phenotypic severity, immune dysfunction, and inflammatory responses were comprehensively assessed. Targeted oxylipin lipidomics was performed to profile lipid mediator alterations, while molecular docking, molecular dynamics simulations, and cellular thermal shift assays (CETSA) were employed to identify and validate the direct target and active components of WZP. RESULTS:WZP treatment significantly ameliorated AD-like symptoms, restored the Th1/Th2 immune balance, reduced mast cell infiltration and cutaneous inflammatory cytokine expression, and lowered serum IgE levels. Notably, psychological stress exacerbated AD by upregulating ALOX15 expression and disrupting the cutaneous oxylipin profile, with marked elevations in pro-inflammatory mediators such as 15-HETE. WZP effectively reversed these stress-induced oxylipin derangements. Mechanistically, cimifugin, a bioactive component of WZP, was identified as a direct inhibitor of ALOX15, binding stably to the enzyme and suppressing its activity, thereby restoring oxylipin homeostasis. CONCLUSION:WZP alleviates stress-aggravated AD by inhibiting ALOX15-mediated oxylipin disruption, with cimifugin as a key active component. This reveals a novel "Stress-ALOX15-oxylipin-inflammation" axis and supports WZP as a promising therapy for stress-associated AD.

OBJECTIVE:To investigate how Huatuo Zaizao pill (HZP) enhances neurological function and to explore the underlying mechanism in acute phase of the middle cerebral artery occlusion (MCAO) ischemia-reperfusion model. METHODS:MCAO ischemia-reperfusion models were established in rhesus monkeys and rats to evaluate cerebral infarct volume and neurological impairment. Additionally, immunofluorescence staining and Western blotting (WB) were utilized to determine the expression levels of glial fibrillary acidic protein (GFAP) and Bax proteins. RESULTS:In the rhesus monkey MCAO ischemia-reperfusion model, the HZP group exhibited a greater reduction in infarct volume than the model group. The NHPSS scores in the HZP group began to decrease at 16 hours after stroke; compared with the model group, the NHPSS scores were significantly lower from 48 hours to 14 days. Compared with the control group, the percentages of cerebral infarction volume in rats of the model group was significantly increased, while that in the HZP groups was significantly decreased. Additionally, the time in the adhesive removal test was significantly shortened in the HZP groups. Immunofluorescence and WB revealed that the expression of GFAP and Bax was increased in the model group, whereas this change could be reversed to a certain extent after HZP treatment. CONCLUSIONS:HZP significantly alleviates acute-phase symptoms in the rhesus macaque MCAO model, with these beneficial effects persisting into the recovery phase. This protective action may counteract cerebral ischemia-reperfusion injury by mitigating neuroinflammation and inhibiting neuronal apoptosis. Findings from both rhesus macaque and rat models in this study support the potential of HZP as an adjunctive therapeutic agent for acute cerebral infarction.

INTRODUCTION:This study aimed to explore the potential neuroprotective effects of Huatuo Zaizao pills (HTZZ) in modulating microglial polarization-mediated neuroinflammation and apoptosis. MATERIALS & METHODS:Twenty-four male mice were included in the study. Sixteen mice underwent permanent middle cerebral artery occlusion (pMCAO) and were randomly assigned to either the MCAO group (vehicle, n = 8) or the MCAO+HTZZ group (HTZZ, 5.80 g/kg, n = 8). Eight mice underwent sham surgery and received vehicle treatment (Sham group). HTZZ or vehicle was administered orally for seven days before surgery and continued for three days after surgery. Neurological functions were evaluated using the Longa score, adhesive removal test, grid-walking test, and forelimb grip strength test. Neuronal damage was assessed via Nissl staining and double immunofluorescence staining for postsynaptic density protein 95 (PSD95) and microtubule-associated protein 2 (MAP2). We also quantified pro-inflammatory cytokines as well as markers of microglial polarization. Apoptosis was assessed through both TUNEL assays and Western blot quantification of key apoptosis-associated proteins. RESULTS:When comparing HTZZ-treated and untreated ischemic animals at 72 h after pMCAO, mice receiving HTZZ exhibited notable functional improvement. HTZZ treatment suppressed pro-inflammatory cytokines and decreased GFAP- and Iba-1-positive cells in the peri-infarct area. Analysis of microglial phenotypes revealed that HTZZ diminished the expression of iNOS/Iba-1+ cells while increasing Arg-1/Iba-1+ cells (both P<0.05). HTZZ markedly lowered the proportion of TUNEL-positive profiles and shifted the balance of apoptosis regulators toward higher Bcl-2/Bax ratios. DISCUSSION:These findings suggest that HTZZ may exert neuroprotective effects through modulation of microglial polarization and attenuation of neuroinflammatory responses. By shifting microglia/macrophages away from a pro-inflammatory phenotype and suppressing cytokine overproduction, HTZZ appears to mitigate secondary neuronal injury in the peri-infarct region. The observed reduction in apoptotic markers and restoration of the Bcl-2/Bax balance indicate HTZZ's potential role in inhibiting neuronal apoptosis. Notably, increased Akt phosphorylation was observed, indicating the possible involvement of the PI3K/Akt pathway. CONCLUSION:HTZZ demonstrates potential neuroprotective effects in permanent cerebral ischemia, possibly through modulation of microglial polarization and attenuation of neuronal apoptosis. These findings support further investigation into HTZZ as a multi-target therapeutic candidate for ischemic stroke.

INTRODUCTION:Cardiovascular Diseases (CVDs) and Gastrointestinal Disorders (GIDs) show significant comorbidity. We established a rat model of chronic atrophic gastritis (CAG) complicated by acute myocardial ischemia (AMI) to investigate the protective effects and mechanisms of Wei Nai An Capsule (WNAC) on CAG and AMI. METHODS:Potential mechanisms were predicted by using network pharmacology and GEO database analysis. To model CAG, we combined intragastric administration of alcohol, sodium deoxycholate, and ammonia. Injections of β-adrenergic agonist isoproterenol modeled the AMI. Histopathological observations were performed, and gastric pH, cardiac marker enzyme levels, cardiac electrical activity, and mRNA expression in the animals were monitored. RESULTS:Network pharmacology and GEO database analyses suggested that WNAC's positive effect on acute myocardial ischemia complicated by chronic atrophic gastritis is associated with the PI3K-AKT signaling pathway. In the model of acute myocardial ischemia complicated by chronic atrophic gastritis, WNAC significantly decreased the elevated ST-segment on the electrocardiogram, reduced gastric pH, reduced levels of markers of cardiac injury, and reduced the mRNA expression of IκBα, PI3K, NF-κB-p65, and COX2 in cardiac and gastric tissues. DISCUSSION:This study demonstrated the crucial need to suppress pro-inflammatory pathways in the treatment of the comorbidity between CVDs and GIDs. However, additional experiments could further strengthen the causal inference. CONCLUSION:This research indicated that WNAC can be successfully implemented to treat the comorbidity of CVDs and GIDs and provides strong evidence for the discovery of new indications of WNAC in clinical practice.

Ethnopharmacological relevance Mammary gland hyperplasia (MGH) is prevalent estrogen-dependent disorder involving hormonal imbalance and psychological comorbidity, which significantly elevates breast cancer risk. Ruhe Sanjie Tablet (RST), a traditional Chinese herbal formula, is clinically effective in alleviating MGH symptoms, yet its underlying mechanisms, particularly concerning associated depressive behaviors, remain insufficiently elucidated. Aim of the study This study investigated the therapeutic effects and integrated mechanisms of RST on MGH in a rat model. Materials and Methods The rat MGH model was established through sequential injections of estradiol benzoate for 25 days and followed by progesterone for 5 days. The effects of RST on MGH were evaluated via mammary nipple morphology and depression-like behaviors (open field, sucrose preference, and forced swim tests). Hematoxylin and eosin (H&E) staining was utilized for pathological examination on mammary and ovarian tissues. Serum concentrations of cytokines (IL-6, IL-1β, TNF-α) and sex hormones (E2, PRL, P, FSH, LH) were measured by ELISA. Protein expressions were confirmed via Western blotting (CDK2, Cyclin E) and immunohistochemistry (ERα, ERβ, Cyclin E1, Ki-67, PR, and PCNA in mammary gland, and GnRH, ERα, and ERβ in hypothalamus). An integrated approach employing network pharmacology, molecular docking, dynamics simulations, and untargeted metabolomics was utilized to identify core targets and metabolites of RST constituents. Results RST significantly reduced mammary nipple diameter/height and alleviated depression-like behaviors (increased exploration, sucrose preference, active coping). It decreased serum pro-inflammatory cytokines and rebalanced sex hormones (lowered E2/PRL, elevated P/FSH/LH). Histopathological improvements in mammary and ovarian tissues were observed. Mechanistically, network pharmacology and molecular docking identified CDK2 as a core target, and Epmedin C exhibiting superior binding affinity and stability with a higher relative content (the third) in RST. Experimental validation confirmed that RST downregulated expression of ER, PR, PCNA, CDK2, and Cyclin E in mammary, indicating a disruption of estrogen signaling and G1/S phase cell cycle progression. Metabolomics revealed that RST mainly modulated 10 differential metabolites, upregulating anti-inflammatory lipids and downregulating pro-inflammatory mediators. Conclusion RST alleviates MGH and its comorbid depressive behaviors through multi-target actions involving hormonal rebalancing, suppression of estrogen receptor signaling, inhibition of CDK2-mediated cell cycle progression, and modulation of inflammatory metabolites.

Background:Zhangyanming Tablets (ZYMT) is a proprietary Chinese medicine containing a variety of traditional Chinese medicines, which can be used to treat a wide range of eye diseases, but its exact effect on diabetic retinopathy (DR) and the specific mechanism are still unclear. This study aims to investigate the ameliorative effects and specific mechanisms of ZYMT on DR. Methods:Key regulatory genes and potential therapeutic targets of ZYMT for DR were evaluated using network pharmacological analysis. Diabetic db/db mice were given low-dose ZYMT (330 mg/kg) and high-dose ZYMT (660 mg/kg), and relevant metabolic indices were tested. Histochemical staining and optical coherence tomography angiography (OCTA) were used to evaluate the histopathological structure of mice retina, RT-qPCR, TUNEL staining and immunofluorescence staining were used to evaluate the anti-apoptosis and anti-angiogenesis effect of ZYMT on DR. Results:The results of network topology analysis showed that the top 10 Traditional Chinese Medicine (TCM) ingredients of ZYMT were quercetin, luteolin, kaempferol, wogonin, naringenin, β-sitosterol, baicalein, isorhamnetin, acacetin, and stigmasterol. ZYMT treats DR through key nodes such as AKT1, TNF, MAPK8, RELA, VEGFA, HIF1A, IL6, CASP3, BCL2, STAT3, and ICAM1. ZYMT has a direct effect on DR rather than secondary improvement of metabolic indices. Tissue staining demonstrated that ZYMT improved retinal vascular morphology and delayed retinal thinning in db/db mice. The OCTA imaging also showed that ZYMT increased blood flow density in db/db mice. TUNEL staining and RT-qPCR results showed that ZYMT could reduce the apoptosis of retinal cells in db/db mice, and RT-qPCR and immunofluorescence staining showed that ZYMT could inhibit retinal neovascularization. Conclusion:This study found the potential target of ZYMT to ameliorate DR through network pharmacological analysis, and verified that ZYMT can improve DR by exerting anti-apoptosis and anti-neovascularization.

Background:Jiangqi Dingchuan Pill (JDP) is a patent Chinese medicine in the treatment of asthma. JDP consists of six herbal drugs, namely, Ephedrae Herba, Mori Cortex, Citri Reticulatae Pericarpium, Perillae Fructus, Descurainiae Semen, Sinapis Semen. Objective:To employ the tools of network pharmacology and in vivo experiments, exploring the possible mechanism of JDP in treating chronic obstructive pulmonary disease (COPD). Materials and Methods:Chemical constituents of JDP, collection of targets of COPD, target prediction were conducted, and then network pharmacological analysis was performed based on protein-protein interaction (PPI). The cigarette smoke and lipopolysaccharide-induced COPD model was applied to assess the effects of JDP. Rats were randomly divided into five groups (n = 8), ie, a sham group, a COPD-control group, two COPD groups treated with different doses of JDP (1.26 and 2.52 g/kg/d, respectively), and one COPD group treated with aminophylline (54 mg/kg/d). Pulmonary functions were assessed. The inflammatory cytokines in bronchial alveolar lavage fluid (BALF) were quantified using enzyme-linked immunosorbent assay (ELISA). The expression of matrix metalloprotein-9 (MMP-9) was quantified using Western blot. Results:A total of 108 genes were found to be the main target genes regulated by JDP in the treatment of COPD, according to PPI analysis. Compared with the COPD-control group, rats in the JDP group exhibited amelioration in lung function, including 20 ms forced expiratory volume/forced vital capacity, maximal mid-expiratory flow curve, and airway resistance (all p < 0.05). A reduction of IL-1β and TNF-α expressions in BALF was also observed (both p < 0.05). Compared with the COPD-control group, the expression of MMP-9 in lung tissue was down-regulated in the JDP group (p < 0.05). Conclusion:This study explored the effects and its mechanisms of JDP in COPD treatment. JDP exhibited therapeutic potential as a COPD intervention drug.

Zishen Yutai Pill (ZYP) is a widely used Chinese patent medicine in the clinical treatment of diminished ovarian reserve (DOR) induced infertility in China. However, the pharmacological actions and underlying mechanisms of ZYP in treating DOR remain poorly understood. This study aimed to evaluate the therapeutic effects of ZYP on DOR and elucidate its potential mechanisms. Two models including tripterygium glycosides (TGs)-induced DOR rat model and pZP3-induced DOR mouse model were selected to assess the effectiveness of ZYP in treating DOR. The intervention lasted for 4 weeks in both models. ZYP administration significantly increased the primordial follicles and the serum levels of AMH both in DOR rat and mouse model. Further, ZYP regulated the apoptosis pathway and apoptosis-related molecules including PI3K-Akt signaling pathway, Bcl6 and Abtb2 in ovary of DOR rat. The ovarian apoptosis level was significantly downregulated in a dose-depended manner in ZYP groups. Thus, we demonstrate that ZYP improved the ovarian reserve in DOR models. The mechanisms of ZYP on DOR may be mediated through decreasing the apoptosis level by the regulation of PI3K-Akt signaling pathway and apoptosis molecules (Bcl6, Abtb2) in ovary.

Ethnopharmacological relevance: The Zishen Yutai pills (ZYP), a Chinese medicinal formulation derived from the Qing Dynasty prescription "Shou Tai pills", have been documented to exhibit beneficial effects in clinical observations treating premature ovarian failure (POF). However, the anti-POF effects and its comprehensive systemic mechanism have not yet been clarified. Aim of the review: Therapeutic effects and systemic mechanism of ZYP in POF were evaluated. Materials and methods: After pulverization, sieving, and stirring, ZYP was administered intragastrically to cisplatin-induced POF mice at a dose of 1.95 mg/kg/d for 14 days. The anti-POF effects of ZYP were investigated by assessing the number of ovarian follicles at different developmental stages, as well as measuring serum estradiol (E2) levels and ovarian-expressed anti-Mullerian hormone (AMH). Reproductive performance and offspring health were evaluated to predict fertility restoration. Furthermore, a combination of proteomic and metabolomic profiling was employed to elucidate the underlying molecular mechanism of ZYP in treating POF. Western blot (WB) analyses and real-time quantitative polymerase chain reaction (RT-qPCR) were conducted to explore the mechanisms through which ZYP exerted its anti-POF effects. Results: We have demonstrated that oral administration of ZYP reversed the reduction in follicles at different developmental stages and stimulated the expressions of serum E2 and ovarian-expressed AMH in a cisplatininduced POF model. Additionally, ZYP ameliorated follicle apoptosis in ovaries affected by cisplatin-induced POF. Furthermore, treatment with ZYP restored the quantity and quality of oocytes, as well as enhanced fertility. Our results revealed 62 differentially expressed proteins (DEPs) through proteomic analyses and identified 26 differentially expressed metabolites (DEMs) through metabolomic analyses. Both DEPs and DEMs were highly enriched in the arachidonic acid (AA) metabolism pathway. ZYP treatment effectively upregulated the protein and mRNA expression of critical targets in AA metabolism and the AKT pathway, including CYP17 alpha 1, HSD3 beta 1, LHR, STAR, and AKT, in cisplatin-induced POF mice. Conclusions: These results indicated that ZYP exerted protective effects against POF and restored fertility from cisplatin-induced apoptosis. ZYP could be a satisfying alternative treating POF.

Jiawei Huoxiang Zhengqi Pill (JHZP) is a commonly used Chinese patent medicine for the clinical treatment of headache, dizziness, chest tightness as well as abdominal distension, and pain caused by wind-cold flu. In this study, a comprehensive strategy combining ultra-high performance liquid chromatography with diode array detector (UHPLC-DAD) fingerprinting and multi-component quantitative analysis was established and validated for quality evaluation of JHZP. A total of 49 characteristic common peaks were selected in a chromatographic fingerprinting study to assess the similarity of 15 batches of JHZP. Furthermore, 109 compounds were identified or preliminarily identified from JHZP by coupling with an advanced hybrid linear ion trap-Orbitrap mass spectrometer. For quantification, the optimized ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) method was employed for the simultaneous determination of 13 target compounds within 12 min. The sensitivity, precision, reproducibility, and accuracy of the method were satisfactory. This validated UPLC-MS/MS method was successfully applied to analyzing 15 batches of JHZP. The proposed comprehensive strategy combining UHPLC-DAD fingerprinting and multi-component UPLC-MS/MS analysis proved to be highly efficient, accurate, and reliable for the quality evaluation of JHZP, which can be considered as a reference for the overall quality evaluation of other Chinese herbal formulations.

Indomethacin, as a non-steroidal anti-inflammatory drugs, is widely used in the clinic. However, it can cause severe injury to the gastrointestinal tract and the incidence is increasing. It has become an essential clinical problem in preventing intestinal damage. Teprenone has been reported to have a significant positive effect on intestinal mucosal lesions, but long-term use of teprenone can elicit adverse reactions. WeiNaiAn capsule is a traditional Chinese medicine formulation used widely in the treatment of gastric and duodenal mucosal injury. However, how WeiNaiAn protects against intestinal mucosal injury and its mechanism of action are not known. In this study, WeiNaiAn capsule or Teprenone treatment improved the intestinal mucosal pathological score and antioxidant level in indomethacin-induced rats. 16 S rRNA sequence data showed WeiNaiAn capsule reverted the structure community and replenished the beneficial bacteria. Furthermore, fingerprint analysis revealed multiple components of WeiNaiAn capsule, including calycosin glucoside, ginsenoside Rg1, ginsenoside Rb1, taurocholic acid sodium, formonetin, and calycosin glucoside. The components of WeiNaiAn capsule promoted the wound healing of the epithelial cell in vitro. Moreover, the components of WeiNaiAn capsule inhibited the protein expressions of phosphoinositide 3-kinase /protein kinase B /mammalian target of rapamycin in hydrogen peroxide or lipopolysaccharides-induced cell model. In conclusion, WeiNaiAn capsule improves intestinal mucosal injury by regulating cell migration, enhancing antioxidant activity, and promoting the structure of the bacterial community homeostasis, the multiple targets provide the parameters for the treatment in the clinic.

Objective: Zishen Yutai Pill (ZYP) is a frequently used traditional Chinese medicine (TCM) preparation in women’s health. However, the effects of ZYP on endometrial epithelial response have not been fully explored. Herein, uterine natural killer cell (uNK) secretion medium was used to mimic the uterine microenvironment. Thereafter, an endometrial epithelial cell line (Ishikawa cells) was treated with ZYP-containing serum to elucidate the effects of ZYP on endometrial receptivity.Methods: uNK cells were isolated from decidual tissues of pregnant women undergoing pregnancy termination surgery, and thereafter, uNK secretion medium was collected. ZYP-containing serum was collected from rats after intragastrical administration of ZYP. Ishikawa cells were divided into three groups, one treated with blank control (control group), one treated with uNK secretion medium (uNK group), and one treated with both uNK secretion medium and ZYP-containing serum (ZYP + uNK group). Total RNAs were extracted. Gene expression profiles of Ishikawa in different groups were determined through microarray analysis. mRNA expressions of selected genes were determined through quantitative real-time polymerase chain reaction (qRT-PCR). Expression of intercellular cell adhesion molecule-1 (ICAM-1) was determined using Western blotting (WB). Results: Compared with the uNK group, the gene expressions of ZYP group with a total of 1117 genes were significantly altered, among which 510 genes were upregulated and 607 genes were downregulated. Compared with uNK group, expressions of CSF1, CSF2, SPP1, and ICAM1 were upregulated (P < 0.05). Up-regulation of ICAM-1 expression after treatment of ZYP was further confirmed by WB analysis. Conclusion: In brief, in the presence of uNK cell medium, ZYP could improve the expressions of ICAM1, CSF1, CSF2, TNF, SPP1, etc. However, further exploration should be carried out in in vivo experiments for the validation of the mechanisms of ZYP on endometrial epithelial response.

目的 建立消渴丸中多糖含量的测定方法.方法 以葡萄糖为对照,采用苯酚-硫酸法对消渴丸中的多糖进行显色,运用紫外-可见分光光度计测定吸光度,计算消渴丸中多糖的含量,并对该方法的精密度、重复性、加样回收率等进行分析.结果 该方法测得消渴丸中多糖含量在0～0.0510 mg/mL的线性范围内,相关系数为0.9999.精密度实验相对标准偏差(relative standard deviation,RSD)为0.90％,重复性实验RSD为1.38％,平均加样回收率为98.77％,RSD为3.25％.结论 应用苯酚-硫酸法测定消渴丸中的多糖含量准确度高、精密度良好、重复性稳定、加样回收率高,适用于消渴丸中多糖含量的测定.

ObjectiveTo assess whether the administration of Zishen Yutai Pill (ZYP) could improve the pregnancy outcomes in different subgroups of women undergoing fresh embryo transfer cycles.Materials and methodsThis is a post hoc analysis of a large scale, placebo-controlled, double blind, randomized clinical trial (RCT) regarding the use of ZYP during assisted reproductive technology (ART) treatment. The RCT was conducted at 19 in vitro fertilization (IVF) centers between April 2014 and June 2017. A total of 2265 women undergoing fresh embryo transfer cycles were randomly assigned in a 1:1 ratio to receive ZYP (n = 1131) or placebo (n = 1134). Post hoc logistic regression analyses were applied in this study to examine the between-group differences of ZYP and placebo on clinical pregnancy rate among different subgroups. Detailed analyses, both in intention-to-treat (ITT) and per-protocol population, were also conducted in specific subgroups with regards to rates of implantation, biochemical pregnancy, clinical pregnancy, live birth, pregnancy loss, as well as other neonatal indices.ResultsZYP showed a significantly higher clinical pregnancy rates than placebo in the ITT population. Detailed subgroup analyses were conducted in subgroup in advanced maternal age (AMA, ≥ 35 years old) and overweight/obese patients (BMI > 24), due to the clinical importance and statistical results. In these subgroups, baseline characteristics were similar between two arms (all P > 0.05). Significantly elevated clinical pregnancy rates were observed in ZYP cohort (both P < 0.05) compared with the placebo group. Results also showed that ZYP treatment resulted in significantly higher rates of implantation, biochemical pregnancy in AMA or overweight/obese patients in ITT analysis (all P < 0.05).ConclusionsThe current post hoc subgroup analysis suggested that AMA and overweight/obese women could experience clinical benefits when treated with ZYP in their fresh embryo transfer cycles. The study provides references for the use of ZYP in ART practices. However, further studies in specific subgroups should be examined in more rigorous clinical trial settings.Clinical trial registrationChictr.org.cn, ChictrTRC-14004494.

Objective To investigate the Xiaoke Pills for hypotensive and hypolipidemic effects on type 2 diabetic db/db mice. Methods By analyzing the regularity of recipe composition,the systolic and diastolic blood pressure,fasting blood glucose, insulin, insulin resistance index（HOMA-IR）, serum total cholesterol（TC）and lowdensity lipoprotein cholesterol（LDL-C）in spontaneously diabetic db/db mice were compared in the low-dose Xiaoke Pills group（1.0 g/kg）, the high-dose Xiaoke Pills group（4.0 g/kg）, the pure Chinese medicine of Xiaoke Pills group（3.996 g/kg）and the Glibenclamide group（4.0 mg/kg）before and after administration. Serum angiotensin Ⅱ（Ang Ⅱ） and hepatic peroxisome proliferator-activated receptor alpha（PPARα） levels were compared between the groups after treatment. Results Compared with the normal control group,mice in the model group had characteristics of type 2 diabetes mellitus with abnormally elevated systolic blood pressure and lipids（P＜0.05 or P＜0.01）. In terms of blood pressure regulation,the systolic blood pressure in the low-and highdose Xiaoke Pills groups and the pure Chinese medicine of Xiaoke Pills group was lower than that in the model group after 4 weeks of administration（P＜0.05）,while there was no significant change in systolic blood pressure in the Glibenclamide group（P＞0.05）. In terms of fasting blood glucose,insulin,HOMA-IR regulation,the effect in the high-dose Xiaoke Pills group was comparable to that in the Glibenclamide group（P＞0.05）,the effect of fasting blood glucose and HOMA-IR in the high-dose group was superior to that in the low-dose group and the pure Chinese medicine of Xiaoke Pills group（P＜0.05）. In terms of blood lipid regulation,the effect of TC and LDL-C levels in the low-and high-dose Xiaoke Pills groups and the pure Chinese medicine of Xiaoke Pills group were comparable to that in the Glibenclamide group（P＞0.05）. The results of Ang Ⅱ and PPARα showed as follows:compared with the model group,the serum Ang Ⅱ level was significantly decreased in both the low-and high-dose groups of Xiaoke Pills and the pure Chinese medicine group（P＜0.05 or P＜0.01）,while there was no significant difference in the Glibenclamide group（P＞0.05）;compared with the model group,the liver PPARαlevel was significantly increased in both the low-and high-dose groups and the pure Chinese medicine group of Xiaoke Pills and the Glibenclamide group（P＜0.05 or P＜0.01）,among which,the PPARα level of liver tissue in the high-dose group of Xiaoke Pills was the highest. Conclusion The efficacy of Xiaoke Pills in type 2 diabetic db/db mice is not only blood glucose regulation,but also hypotensive and hypolipidemic effect,the efficacy of Xiaoke Pills being attributed to the synergistic effect of the pure Chinese medicine part and the western Glibenclamide part.Xiaoke Pills play a role in lowering blood glucose by Glibenclamide regulating HOMA-IR,in regulating blood pressure by the pure Chinese medicine part inhibiting Ang Ⅱ to lower systolic blood pressure,and in regulating blood lipids by Chinese medicine part and western medicine part increasing liver tissue PPARα expression.

Retinitis pigmentosa (RP) is the most common genetic disorder that causes blindness. At present, there exists no remedy for the disease. The aim of the current research was to investigate the protective effect of Zhangyanming Tablets (ZYMT) in a mouse model of RP, and explore the underlying mechanism. Eighty RP mice were randomly divided into two groups. The mice in ZYMT group were administered with ZYMT suspension(0.0378 g/mL), while the mice in model group were given the same volume of distilled water. At day 7 and day 14 after intervention, electroretinogram (ERG), fundus photography, and histological examination were used to assess the retinal function and structure. TUNEL, immunofluorescence and qPCR were used to evaluate cell apoptosis and expressions of Sirt1, Iba1, Bcl-2, Bax and Caspase-3. A significantly shortened latency of ERG waves was observed in ZYMT-treated mice, in comparison to those in the model group (P < 0.05). Histologically, ultrastructure of the retina was better preserved, and the outer nuclear layer (ONL) exhibited marked increase in thickness and cell count in ZYMP group (P < 0.05). The apoptosis rate was decreased markedly in ZYMT group. Immunofluorescence analysis showed that the expressions of Iba1 and Bcl-2 in the retina were increased, Bax and Caspase-3 were decreased after ZYMT intervention, while the qPCR revealed that the expressions of Iba1 and Sirt1 were significantly increased (P < 0.05). This study indicated that ZYMT has protective effect on retinal function and morphology of inherited RP mice in the early stage, possibly mediated via the regulation of antioxidant and anti-/pro-apoptotic factors expressions.

Objective:Zishen Yutai Pill (ZYP), containing 15 Chinese traditional medicine, is a safe and well quality-controlled TCM preparation with promising effects in many fields of reproduction. The current study was designed to investigate the therapeutic effects of ZYP on sperm quality and testis in varicocele (VC) rats. Materials and methods:Male Wistar rats were randomly divided into four groups (n = 6), i.e., a sham group, a VC group, and VC groups treated with different dose of ZYP (1575 and 3150 mg/kg/d, respectively). The experimental VC model was established by partial ligation of left renal vein. Six weeks after model establishment, ZYP was orally administered once a day for the next 6 weeks. Parameters relating to testis and sperm quality were assessed. Hematoxylin-eosin staining was used to showed testicular tissue damage in experimental VC rats. Expressions of proteins relating to NLRP3 inflammasome pathways were determined using Western blot (WB). The mRNA expressions of relating genes were determined using quantitative real-time PCR (qRT-PCR) analysis. Results:ZYP could significantly improve sperm motility and decrease sperm DNA fragmentation index in VC rats (P < 0.05). Hematoxylin-eosin (HE) staining showed that ZYP could alleviate testicular tissue damage caused by experimental varicocele in rats. Compared to the VC model, expressions of NLRP3, ASC, and caspase-1 in rats treated with ZYP were significantly down-regulated, as validated by both qRT-PCR and WB analysis (P < 0.05). Conclusions:In brief, ZYP could improve sperm DNA integrity by inhibiting the NLRP3 inflammasome pathway and alleviating the chronic inflammation of testicular tissue induced by experimental varicocele in rats.