ETHNOPHARMACOLOGICAL RELEVANCE:Guilu Erxian Oral Liquid (GLEX) is a traditional Chinese medicinal formula first recorded in Yi Bian, a medical classic written during the Ming Dynasty, and has been conventionally used to treat male infertility based on its kidney tonification and is now widely recognized for its therapeutic effects against oligoasthenospermia (OAS). AIM OF THE STUDY:This study aims to investigate the impact of GLEX on cyclophosphamide (CTX)-induced OAS-like mice and to explore its underlying mechanisms, utilizing pharmacodynamics, multi-omics, and molecular biological techniques. MATERIALS AND METHODS:The chemical constituents of GLEX were identified using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The binding affinity between GLEX constituents and DJ-1 was determined by molecular docking and molecular dynamics simulations. A mouse model of OAS was induced using 60 mg/kg CTX for 5 days. ICR mice were randomly divided into seven groups: Control group, Model group, low (1.93 mL/kg)-, medium (3.86 mL/kg)-, and high (7.72 mL/kg)-dose groups of GLEX, Wu Zi Yan Zong pill group, and L-Carnitine group. Each group received consecutive gavage for 28 days. Semen quality parameters were assessed using a sperm quality analyzer, along with morphological assessment by hematoxylin and eosin (HE) staining. Serum sex hormone levels were measured using an enzyme-linked immunosorbent assay (ELISA). Gut microbiota was profiled via 16S rRNA gene sequencing. RNA-Seq and DIA proteomics were used to screen the potential pathways and targets associated with OAS pathogenesis and GLEX treatment. Subsequently, the ultrastructure of testis was examined under a transmission electron microscope. Testicular oxidative stress markers and Na+-K+-ATPase activity were measured by biochemical analysis and fluorescence microscopy. The Mitochondria-Complement-NETs network targets (DJ-1, UQCRQ, C3, FGG, FGA, NE-DNA, MPO-DNA, citH3, and PAD4) were assessed by immunohistochemistry detection, immunofluorescence staining, ELISA, and western blotting. RESULTS:A total of 84 components were detected by UPLC-MS/MS analysis. After merging with the 15 components predicted from the BATMAN-TCM database and screening against the TCMSP database, 13 were structurally identified. Molecular docking and molecular dynamics simulations revealed a stable binding between GLEX constituents and DJ-1. CTX-induced OAS-like mice showed markedly reduced sperm density and motility and imbalanced serum sex hormone levels, along with structural damage in testicular and epididymal tissues. GLEX effectively restored these alterations. Besides, GLEX remodeled the gut microbiota composition and function in OAS mice. Multi-omics analyses further revealed that the therapeutic mechanisms of GLEX were associated with mitochondria, energy metabolism, oxidative phosphorylation, complement and coagulation cascades, and neutrophil extracellular trap (NET) formation pathways. OAS mice performed disrupted testicular mitochondrial ultrastructure, decreased activities of Na+-K+-ATPase, T-SOD, GSH-Px, CAT, DJ-1, and UQCRQ, and elevated levels of ROS, MDA, C3, FGA, FGG, MPO-DNA, NE-DNA, citH3, and PAD4. GLEX significantly reversed them via the Mitochondria-Complement-NETs network, exhibiting mitochondrial protection, complement inhibition, and NETs blockade. CONCLUSION:GLEX exerts protective effects on semen quality, endocrine function, and histopathological injury of CTX-induced OAS-like mouse model, accompanied by the coordinated modulation of mitochondrial function, complement overactivation, and NETs release. These findings delineate a proposed Mitochondria-Complement-NETs network, which warrants further investigation and may inform future research on interventions for chemotherapy-related reproductive impairment.

Objective To predict the mechanism of action of Laiyang Pear Cough Syrup in treating cough by using network pharmacology, molecular docking technology and the predicted key pathways in animal experiments. Methods Chemical components of each medicinal flavor were collected and screened from TCMSP and other databases to establish a database of the components of Laiyang Pear Cough Syrup. Cough-related targets were obtained from TTD and other databases. A Venn diagram of the components targets and cough targets was drawn to prelim determine the potential targets of the active ingredients of Laiyang Pear Cough Syrup in the treatment of cough. A PPI network was constructed and key targets were screened. The data were into Cytoscape 3.10.1 software, and a "active component-disease target" network was constructed, and key components were screened. The screened targets were subjected to GO enrichment analysis and KEGG pathway enrichment analysis. Molecular docking verification between key targets and core active ingredients was conducted using AutoDock Tools 1.5.6 software. A citric acid-induced guinea pig cough model was established. The cough latency and cough frequency were measured in the control and treatment groups, and the PI3K-AKT pathway predicted by network pharmacology was validated via Western blot analysis. Results 43 active chemical components, 76 common targets, 591 GO biological processes, and 159 KEGG signaling pathways were screened out. The active components of Laiyang Pear Cough Syrup in the treatment of cough may be quercetin, stigmasterol, and luteolin, and the key may be IL-6, TNF, AKT1, IL-1β, and PTGS2, which mainly regulate the PI3K/AKT pathway to relieve cough. Molecular docking revealed that luteolin, quercetin, and stigmasterol as the key active ingredients in Laiyang Pear Cough Syrup bind to AKT1 with high affinities, displaying docking scores of −8.1, −8.0, and −7.7 kcal mol−1. These results indicate that the compounds may function as potent agonists to activate the PI3K-AKT signaling pathway. Laiyang Pear Cough Syrup can significantly prolong the cough latency period and reduce the number of coughs (compared with the control group, P < 0.01 or 0.05), and down-regulate the expression of PI3K, p-PI3K, AKT, p-AKT proteins. Conclusion Laiyang Pear Cough Syrup can relieve cough through a variety of components, targets and pathways. Regulating the PI3K-AKT pathway may be its main mechanism of action.

BACKGROUND:As the global population ages, the burden of drug use disorders (DUDs) among the elderly is rising. It is imperative to conduct a quantitative analysis of the disease burden affecting this vulnerable population. METHODS:Utilize the Global Burden of Disease Study 2021 database to obtain incidence rates and disability-adjusted life years (DALYs) for opioids, cocaine, amphetamines, and cannabis among the elderly (aged 60-89) across 204 countries and 5 SDI regions from 1990 to 2021. Employ Joinpoint regression analysis to calculate the average annual percentage change (AAPC) of age-standardized incidence rates (ASIR) and age-standardized DALYs rates (ASDR). Use the Das Gupta method to decompose and analyze the impacts of changes in age structure, population growth, and epidemiology on DALYs during this period. Finally, apply the Nordpred model to predict DALYs for global and high-burden regions from 2022 to 2035. RESULTS:Of the four DUDs, opioids have the highest disease burden. Joinpoint analysis indicates that from 1990 to 2021, the ASIR and ASDR remained stable. Cocaine use disorder ASIR remained stable, but ASDR increased with an AAPC of 0.94 (95% CI 0.77-1.11). The burden of amphetamine and cannabis use disorders generally stabilized. Geographic heterogeneity was evident at regional and national levels, with ASDR for all four DUDs increasing in high-SDI areas while remaining stable or declining in other SDI areas. High-income North America, represented by the United States, shows a higher burden of disease. Decomposition analysis shows that population growth is the main factor affecting the change in the burden of DUDs in most regions, and high-income North America is mainly affected by epidemiological changes. According to the Predictive models, the DALYs of DUDs in the global elderly population is still on the rise, especially in the man group in North America. CONCLUSION:The burden of DUDs among the elderly varies across countries, regions, SDI levels, and genders, underscoring the need for targeted public health policy adjustments and strategic allocation of medical resources to mitigate this burden.

The pathogenesis of diabetic retinopathy DR is highly complex, and in recent years, increasing attention has been directed toward the role of the gut in its development. Evidence suggests that intestinal barrier dysfunction is a frequent pathological feature in both DR patients and animal models, facilitating the translocation of gut microbiota and their metabolites into the systemic circulation. On one hand, these translocated microbial components and metabolites activate intestinal immune cells, promoting the migration of gut-derived immune cells to the retina or choroid, thereby contributing to retinal neuroinflammation. On the other hand, they may compromise the integrity of the blood-retina barrier, disrupting the retinal microenvironment. This review explores the influence of the gut on DR pathogenesis through several key mechanisms: alterations in gut microbiota and their metabolites, impairment of intestinal barrier integrity, dysregulated gut immune responses, translocation routes of gut-derived components (including immune cells, microbes, and metabolites) to the retina, and the resulting retinal pathology and microglial-mediated neuroinflammation. By elucidating the pathological mechanisms underlying the gut-retina axis, we aim to identify critical molecular and cellular targets involved in DR progression and to provide new insights for the development of innovative therapeutic strategies.

Oxidative stress is a major contributor to male infertility, particularly oligoasthenozoospermia. This study aimed to investigate the cytoprotective mechanism of Guilu Erxian Oral Liquid (GLEX) against H₂O₂-induced oxidative damage in spermatogonial cells, focusing on miR-6739-5p regulation and activation of the PI3K/AKT pathway using histocytochemical approaches. An oxidative stress model was established in rat spermatogonial stem cells (SSCs) with 250 µM H₂O₂. Cell proliferation, apoptosis, reactive oxygen species (ROS) accumulation, and DNA oxidative damage were assessed using EdU incorporation, flow cytometry, immunofluorescence, and 8-hydroxy-2'-deoxyguanosine (8-OHdG) ELISA. Expression of miR-6739-5p and Phosphatidylinositol 3-Kinase/Protein Kinase B (PI3K/AKT) pathway components (PIK3CA, p-PI3K, p-AKT) was evaluated by RT-qPCR and Western blotting. The interaction between miR-6739-5p and PIK3CA was confirmed via dual-luciferase reporter assay. The cytoprotective effects of GLEX were examined through pre-treatment and quantified using histochemical and cytological markers. H₂O₂ treatment significantly impaired cell viability, increased apoptosis and ROS production, and upregulated miR-6739-5p. Overexpression of miR-6739-5p exacerbated damage, while silencing reversed it and restored PI3K/AKT signaling. GLEX pretreatment effectively reduced miR-6739-5p expression, restored cell viability, suppressed oxidative and inflammatory markers (ROS, 8-OHdG, TNF-α, IL-1β), and enhanced PI3K/AKT activation. These effects were comparable to PI3K pathway activation. GLEX confers histocytochemical protection to spermatogonial cells under oxidative stress by downregulating miR-6739-5p and activating the PI3K/AKT pathway. This study highlights a novel regulatory mechanism and supports GLEX as a potential therapeutic agent for oxidative stress-associated male infertility.

Cordyceps polysaccharide (CSP) has been shown to exhibit anti-inflammatory and antioxidant effects, with potential applications in ischemic stroke. This work is to explore the interventional potential of CSP in MCAO rats and the effects on the intestinal and cerebral IL-23/IL-17 axis. We conducted pharmacological experiments and mechanism exploration in MCAO rats. Our research showed that CSP improved the neurological function and cerebral pathological morphology, reduced cerebral infarction volume and water content in MCAO rats. We also found that CSP significantly decreased the IL-1 beta, TNF-alpha and IL-6 in the ischemic brain and enhanced the ability of MCAO rats to resist oxidative stress. Additionally, CSP improved intestinal barrier, inhibited the activation of the TLR4/Myd88/NF-kappa beta signaling pathway and IL-23/IL-17 axis. The study results demonstrated the effectiveness of CSP in interfering with MCAO rats. The mechanism appears to be related to protecting the intestinal barrier and inhibiting the IL-23/IL-17 axis.

Ethnopharmacological relevance: Diminished ovarian reserve (DOR) was considered a refractory reproductive endocrine condition that negatively affected female reproductivity. Yangjing Zhongyu Decoction (YJZYD) had effects on treating infertility. However, there were few studies on the mechanisms of YJZYD preserving ovarian reserve.Aim of the study: To explore the possible mechanisms of YJZYD against DOR by UPLC-ESI-MS/MS, network pharmacology, and experimental validation.Methods: The chemicals of YJZYD were measured by UPLC-ESI-MS/MS. The correlating targets of YJZYD and DOR were identified by the ETCM database, GeneCards database, and PubMed database. The common targets were employed with the DAVID database and visualized with the PPI network. GO and KEGG enrichment analyses were carried out to explore biological progression and pathways. In vivo experiments, energy production was assessed by ATP, and apoptosis rate was analyzed by TUNEL. The serum FSH, AMH, and E2 levels were evaluated by ELISA. Western blotting and immunohistochemistry were used to measure the expression of SIRT1, PGC1 & alpha;, NRF1, COX IV, FSHR, CYP19A1, PI3K, p-Akt, Akt, Bcl-2, and Bax.Results: 132 components in YJZYD were identified by UPLC-ESI-MS/MS. 149 overlapped targets were extracted from YJZYD and DOR, and the top 20 common targets included AKT1 and CYP19A1. ATP binding was involved in GO analysis. In the KEGG enrichment analysis, the metabolic pathway was the top, and the PI3K-Akt signaling pathway was included. In vivo experiments, YJZYD improved ovarian index and histomorphology. After YJZYD treatment, serum FSH, E2, and AMH were well-modulated, and the content of ATP was up-regulated. Besides, the expression of Bax was suppressed in ovarian tissue, while the expressions of SIRT1, PGC1 & alpha;, NRF1, COX IV, FSHR, CYP19A1, PI3K, Bcl-2, and p-Akt/Akt were enhanced.Conclusion: YJZYD could attenuate reproductive endocrine disturbance and ovarian lesions in vivo by mediating steroidogenesis, energy metabolism, and cell apoptosis. This study uncovered the mechanisms of YJZYD against DOR, providing a theoretical basis for further study.

Objective: To investigate the mechanism of action of Wuzi Yanzong pill (WYP) in rats with oligoasthenozoospermia (OAZ) via metabolomics and to provide a possible basis for improving this WYP-based treatment. Methods: A rat model of OAZ was established by treating male Sprague–Dawley rats with glucosides from Tripterygium wilfordii Hook. F. Seventy-two rats were randomly divided into six groups: control, L-carnitine (positive control), model, and low-, medium-, and high-dose WYP groups. Rats in the experimental groups were treated with WYP for 4 weeks. At the end of the treatment period, sperm cell quality (density, motility, and viability) was assessed using a semen analysis system, mitochondrial membrane potential (MMP) was assessed using flow cytometry, and testicular injury was assessed using hematoxylin and eosin staining to validate the therapeutic effect of WYP in OAZ. Further, serum metabolomics-based analysis was performed using high-performance liquid chromatography-mass spectrometry to identify differential metabolic pathways and possible mechanisms of action of WYP in OAZ treatment. Results: A rat model of OAZ was considered successfully-established after comparing the quality of spermatozoa in the model group to that in the control group. WYP-M and WYP-H treatments significantly improved sperm cell density, motility, and viability compared with those in the model group (all P < .05). Compared with the model group, both WYP-M and WYP-H treatments increased MMP values (P = .006 and P = .021 respectively), while there was no significant difference in the L-carnitine group. L-carnitine and WYP administration reversed damage to the testes to varying degrees compared with that in the model group. Further, 44 differential metabolites and four metabolic pathways, especially autophagy pathway, related to OAZ were identified via metabolomics. Conclusions: WYP improves sperm cell quality and MMP in OAZ primarily via autophagy regulation. These findings can be employed to improve the efficacy of WYP in humans.

Cefminox sodium carbon nanodots were synthesized via a rapid one-step microwave method using cefminox sodium as sole precursor. The prepared cefminox sodium carbon nanodots had good water solubility and the diameter was concentrated at 1.29 +/- 0.40 nm with relative uniform. They exhibited fluorescence excitation-dependent property, of which optimal excitation wavelength was similar to 390 nm, and optimal emission wavelength was similar to 465 nm. As retained part of the structure and antibacterial properties of cefminox sodium, cefminox sodium carbon nanodots had low biological toxicity and sufficient antibacterial activity against bacteria in vitro. In vivo experiments demonstrated cefminox sodium carbon nanodots could exert a good therapeutic effect on acute bloodstream infection. In the mixed solution (Escherichia coli, blood and cefminox sodium carbon nanodots), the fluorescent intensity exhibited quadratic functional relationship with the concentration of Escherichia coli. The detection concentration range for Escherichia coli was 0.5 x 10(6) similar to 1 x 10(9) CFU/mL, and the detection limit was 3.7 x 10(5) CFU/mL. It suggested cefminox sodium carbon nanodots can rapidly and quantitatively detect the number of Escherichia coli in blood while effectively treating bloodstream infection. Due to the long period of clinical detection time (about 1-5 days or more) for bacterial counts in the blood of patients, this method provides a new strategy for rapid clinical treatment and effective monitoring of the degree of infection in patients.

目的 系统评价复方玄驹胶囊(CXC)治疗慢性前列腺炎(CP)的临床有效性与安全性.方法 检索中国知网、万方数据库、维普数据库、PubMed中CXC治疗CP的随机对照试验,检索时限自2007年1月至2022年8月29日.由两名研究人员按照上述方法进行文献查阅,并对文献进行纳入和排除的筛选.采用RevMan 5.4软件进行数据分析.结果 共纳入21篇文献,累计3401例样本.Meta分析结果显示,CXC治疗后总有效率明显升高(RR=1.32,95％CI为1.26～1.38,P<0.00001),美国国立卫生研究院慢性前列腺炎症状指数(NIH-CPSI)显著降低(MD=-6.16,95％CI为-7.56～-4.76,P<0.00001),但对不良反应无显著影响(RR=0.82,95％CI为0.50～1.33,P=0.42).结论 基于本研究,CXC可改善CP症状,对CP具有显著的临床疗效,但仍需更多高质量、大样本的随机对照试验进一步验证.

AimsRheumatoid arthritis (RA) is a common chronic immune disease. Berberine, as its main active ingredient, was also contained in a variety of medicinal plants such as Berberaceae, Buttercup, and Rutaceae, which are widely used in digestive system diseases in traditional Chinese medicine with anti-inflammatory and antibacterial effects. The aims of this article were to explore the therapeutic effect and mechanism of berberine on rheumatoid arthritis.MethodsCell Counting Kit-8 was used to evaluate the effect of berberine on the proliferation of RA fibroblast-like synoviocyte (RA-FLS) cells. The effect of berberine on matrix metalloproteinase (MMP)-1, MMP-3, receptor activator of nuclear factor kappa-Β ligand (RANKL), tumour necrosis factor alpha (TNF-α), and other factors was determined by enzyme-linked immunoassay (ELISA) kit. Transcriptome technology was used to screen related pathways and the potential targets after berberine treatment, which were verified by reverse transcription-polymerase chain reaction (RT-qPCR) and Western blot (WB) technology.ResultsBerberine inhibited proliferation and adhesion of RA-FLS cells, and significantly reduced the expression of MMP-1, MMP-3, RANKL, and TNF-α. Transcriptional results suggested that berberine intervention mainly regulated forkhead box O (FOXO) signal pathway, prolactin signal pathway, neurotrophic factor signal pathway, and hypoxia-inducible factor 1 (HIF-1) signal pathway.ConclusionThe effect of berberine on RA was related to the regulation of RAS/mitogen-activated protein kinase/FOXO/HIF-1 signal pathway in RA-FLS cells.Cite this article: Bone Joint Res 2023;12(2):91–102.

Bidirectional communication between central nervous system (CNS) and intestine is mediated by nerve, endocrine, immune and other pathways in gut-brain axis. Many diseases of CNS disturb the homeostasis of intestine and gut microbiota. Similarly, the dysbiosis of intestinal and gut microbiota also promotes the progression and deterioration of CNS diseases. IL-23/IL-17 axis is an important inflammatory axis which is widely involved in CNS diseases such as experimental autoimmune encephalomyelitis (EAE), multiple sclerosis (MS), and ischemic stroke (IS). Attributing to the long anatomically distances between ischemic brain and gut, previous studies on IL-23/IL-17 axis in IS are rarely focused on intestinal tissues. However, recent studies have found that IL-17+T cells in CNS mainly originate from intestine. The activation and migration of IL-17+T cells to CNS is likely to be affected by the altered intestinal homeostasis. These studies promoted the attention of IL-23/IL-17 axis and gut-brain axis. IS is difficult to treat because of its extremely complex pathological mechanism. This review mainly discusses the relationship between IL-23/IL-17 axis and IS from the perspective of gut-brain axis. By analyzing the immune pathways in gut-brain axis, the activation of IL-23/IL-17 axis, the roles of IL-23/IL-17 axis in gut, CNS and other systems after stoke, this review is expected to provide new enlightenments for the treatment strategies of IS. This article is part of the Special Issue on "Microbiome & the Brain: Mechanisms & Maladies".

目的 探究鼠李糖乳杆菌R9639、干酪乳杆菌Zhang和植物乳杆菌P9三株益生菌组成的复方益生菌粉对小鼠免疫力的影响.方法 将小鼠分为阴性对照组、低剂量组、中剂量组和高剂量组,每组各10只,并分别灌胃给予同等纯净水,0.21,0.42,1.25 mg/(kg·d)复方益生菌,观察复方益生菌对小鼠体重、脏器/体重比、溶血素半数溶血值、抗体生成细胞数量、迟发型变态反应、淋巴细胞转化、碳廓清功能、腹腔巨噬细胞吞噬功能及NK细胞活性的影响.结果 与阴性对照组比较,低、中、高剂量组小鼠体重、胸腺/体重比值和脾脏/体重比值差异均无统计学意义(P＞0.05);高剂量组小鼠的迟发型变态反应程度显著增加(P＜0.01);低、中、高剂量组小鼠的抗体生产细胞数量、溶血素半数溶血值、淋巴细胞增殖能力、NK细胞活性、碳廓清功能和腹腔巨噬细胞吞噬能力均显著增加(P＜0.01).结论 复方益生菌粉能提高小鼠免疫力,结果符合《保健食品检验与评价技术规范》中"具有增强免疫力功能"的评价标准.

Objective:Paridis Rhizoma is recorded as a minor poison in Chinese Pharmacopoeia, but its toxic mechanism has not been clarified.With the attention paid to the safety of Chinese medicine in recent years, the safety and mechanism of Paridis Rhizoma in clinical use also need to be clarified.The purpose of this paper is to clarify the hepatotoxicity mechanism of Paridis Rhizoma extract.Methods:The acute hepatotoxicity was verified by zebrafish.The zebrafish with 4 days of post-fertilization（4 dpf） were exposed to Paridis Rhizoma extract, acridine orange staining was carried out after 24 h, and the hepatotoxicity was proved by histopathological section.The hepatotoxicity targets of Paridis Rhizoma were predicted by network pharmacology and molecular docking technology, and real-time polymerase chain reaction（RT-PCR） was used to verify the network prediction targets and pathways.Results:According to the experimental results, yellow-green fluorescent spots appeared in the liver of zebrafish in the Paridis Rhizoma water extract group and alcohol extract group after exposure treatment for 24 h.Histopathological sections showed that the arrangement of hepatocytes was disordered and loose, and nuclear apoptosis and vacuoles appeared, indicating hepatotoxicity.The results of network pharmacology predicted that the hepatotoxicity of Paridis Rhizoma may be related to the Ras signaling pathway of cell proliferation, phosphoinositide 3-kinase/protein kinase B（PI3K/Akt） signaling pathway, mitogen-activated protein kinase（MAPK） signaling pathway, and Janus kinase-signal transducer and activator of transcription（JAK/STAT） signaling pathway related to energy metabolism and signal transduction.Vascular endothelial growth factor A（VEGFA）,platelet-derived growth factor receptor beta（PDGFRB）,murine double minute 2（MDM2）,signal transducers and activators of transcription 3（STAT3）,mammalian target of rapamycin（MTOR）,MET,Bcl-2-like protein 1（BCL2L1）,fibroblast growth factor 2（FGF2）,interleukin-2（IL-2）,and MAPK14 may be the core targets of hepatotoxicity caused by Paridis Rhizoma.The results of RT-PCR experiments verified the above speculation.Conclusion:Paridis Rhizoma may affect the signal transduction of hepatocyte proliferation and energy metabolism, which leads to hepatotoxicity.

OBJECTIVE:To explore the mechanism of Shenrong pills in improving oligoasthenospermia by inhibiting oxidative stress-induced Leydig cell apoptosis in mouse testis.METHODS:The oxidative stress model of mouse Leydig cells (TM3) was induced by 3% hydrogen peroxide (H2O2) of 600 μmol/L, and then TM3 cells were treated with 7.5%, 10% and 12.5% serum containing Shenrong pills, respectively. TM3 cells were divided into normal control group, model group, and low, medium and high dose groups of Shenrong pills containing serum. The cell viability of TM3 after treatment was detected by CCK-8 method, reactive oxygen species (ROS) were detected by DCFH probe method, and SOD-1, CAT, GSH-px, MDA and LPO in cell lysates were detected by ELISA method. The changes of apoptosis-related proteins Bcl-2, Bax in cell lysates were detected by Western Blot.RESULTS:After H2O2 treatment, compared with normal control group, cell viability was significantly decreased (P< 0.01), MDA and LPO contents were significantly increased (P<0.01), SOD-1, CAT and GSH-px contents were significantly decreased (P<0.01). Reactive oxygen species (ROS) were significantly increased, the relative expression ratio of Bcl-2 protein was significantly decreased (P<0.01, P<0.05), and the relative expression of Bax protein was increased. After the administration of Shenrong pills containing serum, the above indexes were reversed to varying degrees.CONCLUSION:Shenrong pills can resist oxidative stress, inhibit the apoptosis of TM3 cells in mice, maintain high levels of testosterone required for spermatogenic cells, and improve the sperm quality of mice with oligonasthenospermia.

Ischemic stroke (IS) has complex pathological mechanisms, and is extremely difficult to treat. At present, the treatment of IS is mainly based on intravenous thrombolysis and mechanical thrombectomy, but they are limited by a strict time window. In addition, after intravenous thrombolysis or mechanical thrombectomy, damaged neurons often fail to make ideal improvements due to microcirculation disorders. Therefore, finding suitable pathways and targets from the pathological mechanism is crucial for the development of neuroprotective agents against IS. With the hope of making contributions to the development of IS treatments, this review will introduce (1) how related targets are found in pathological mechanisms such as inflammation, excitotoxicity, oxidative stress, and complement system activation; and (2) the current status and challenges in drug development.

目的:利用网络药理学和分子对接技术探究核心组方治疗精液不液化的作用机制.方法:利用中药系统药理学数据库与分析平台(TCMSP)和Uniprot数据库对王琦院士的核心组方药物进行成分和靶点的处理收集,再借助Gene Cards数据库、在线人类孟德尔遗传数据库(OMIM)对疾病靶点进行收集,与药物靶点取"药物-疾病"交集,获得核心靶点,使用String数据库绘制核心靶点蛋白质-蛋白质相互作用(PPI)网络.运用R语言平台进行基因本体(GO)和京都基因和基因组百科全书(KEGG)信号通路富集分析,构建核心组方"成分-靶点-疾病"通路图.对关键成分与核心靶点进行分子对接,最后通过造模和药物对大鼠进行干预后,利用酶联免疫吸附试验(ELISA)检测各组大鼠前列腺中细胞因子水平,实时荧光定量-聚合酶链反应(RT-PCR)法检测IL-17mRNA的表达量,对核心作用靶点及信号通路进行动物实验验证.结果:核心组方中有活性成分29个,对应靶点194个,与疾病靶点交集靶点24个,关键成分有槲皮素、山柰酚、β-谷甾醇等,核心靶点涉及肿瘤坏死因子(TNF)、纤溶酶原激活物抑制因子(SERPINE1)、白细胞介素6(IL-6)等.主要通过糖尿病并发症AGE-RAGE信号通路及IL-17炎症通路等发挥药效.核心组方中的槲皮素、β-谷甾醇等核心靶点之间具有较强的结合活性.核心组方能显著降低模型组大鼠前列腺组织中TNF、SERPINE1、IL-6、IL-1β的水平及IL-17 mRNA的表达量(P<0.01).结论:王琦院士所拟核心组方中的关键成分槲皮素、β-谷甾醇等可能通过作用于TNF、SERPINE1等靶点调节多条通路,可能通过抑制炎症反应、抗氧化和调节纤维蛋白等作用治疗精液不液化.

Objective: This study investigated the COVID-19-prevention knowledge and practices of healthcare workers (HCWs), their psychological states concerning the return to work, and their trust and requirements in using traditional Chinese medicine (TCM) to prevent and treat COVID-19. It is hoped that the study can serve as a reference for policy making during the resumption of work in other countries or regions experiencing similar situations. Methods: This study comprised a quantitative cross-sectional online survey design. Purposive sampling and Cluster sampling were used to recruit all HCWs working in public hospitals in Huangzhou District, Huanggang City, Hubei Province, China. From April 23 to May 14, 2020, surveys were sent electronically to all 13 public hospitals in this area. Results: In total, 2,079 responses were received and 2,050 completed forms were included. After analysis, 47.9 and 46.6% of HCWs indicated that they possessed very good knowledge or good knowledge of preventative measures, respectively. Multivariable log-binomial regression indicated that male, tertiary hospital, medical staff, and undergraduate/postgraduate qualification were associated with good knowledge. Good knowledge was also well-correlated with good practice (OR: 3.277; 95% CI: 2.734–3.928; P < 0.01). 59.8% of HCWs reported worries about resuming work; especially asymptomatic infections. The Self-Rating Anxiety Scale (SAS) indicated that 10.8% of participants had mild anxiety, 1.5% moderate anxiety, and 0.1% severe anxiety. Female, divorced/widowed, and working in a high risk hospital (the Huangzhou District People's Hospital was used for throat swab examinations of returning workers) were risk factors for concerns about resuming work and anxiety symptoms. However, good preventive knowledge was a protective factor for anxiety. HCWs' trust in using TCM to treat COVID-19 was significantly higher than their trust in using TCM for prevention (P < 0.001). Regarding preferences for preventative TCM products, oral TCM granules were the most preferred (62.4%). HCWs also indicated they wanted to know more about the clinical efficacy, applicable population, and adverse reactions of preventative TCM products (89.3, 81.1, and 81.4%, respectively). Conclusion: While HCWs had good knowledge of COVID-19 preventative measures, this did not eliminate the psychological impact of resumption of work. Promotion of COVID-19 prevention knowledge reduces the risk of infection, and alleviates the worries and anxiety symptoms of HCWs about resuming work (especially in administrative staff, those with low education, and those working in primary hospitals). Additional psychological support is required for female HCWs, divorced/widowed HCWs, and those working in high-risk hospitals. Finally, systematic trials of preventative TCM products are recommended.

Male infertility is a major and growing health problem with an estimated global prevalence of 4.2%. The current therapy is limited by the unknown etiology of MI, emphasizing the critical requirement forward to a more efficient method or medication. Through thousands of years, Traditional Chinese Medicine (TCM) has been shown to be effective in treating MI effectively. However, the components, mechanisms and functions of TCM prescriptions on MI are still obscure, severely limiting its clinical application. In order to discover the molecular mechanism of TCM against MI, our study presents a comprehensive approach integrated data mining, network pharmacology, molecular docking, UHPLC-Q-Orbitrap HRMS, and experimental validation. Here, we begin to acquire 289 clinical TCM prescriptions for MI from a TCM hospital's outpatient department. Then, Core Chinese Materia Medica (CCMM) was then retrieved from the TCM Inheritance Support System (TCMISS), which was utilized to discover the underlying rules and connections in clinical prescriptions. After that, 98 CCMM components and 816 MI targets were obtained from ten distinct databases. Additionally, the network pharmacology methods, including network construction, GO and KEGG pathway enrichment, PPI analysis, were utilized to reveal that kaempferol, quercetin, isorhamnetin, and beta-sitosterol are the core components of CCMM in treating MI. The mechanisms and functions of CCMM against MI are hormone regulation, anti-apoptosis, anti-oxidant stress, and anti-inflammatory. Furthermore, the strong connections between four core components and six key targets were verified using a molecular docking method. Following that, the core components of the CCMM extract were identified using UHPLC-Q-Orbitrap HRMS analysis. Finally, in vivo experiments demonstrated that CCMM and four core components could improve the density, motility, viability of sperm, lecithin corpuscle density, decrease the rate of sperm malformation and testis tissue damage, and regulate the protein expressions of AKT1, MAPK3/1, EGFR, and TNF-α in a mouse model of MI. UHPLC-Q-Orbitrap HRMS analysis and in vivo experiments further validated the results of data mining, network pharmacology, and molecular docking. Our study could uncover the components, mechanisms, and functions of TCM prescriptions against MI and develop a new integrative approach to demonstrate TCM's multi-component, multi-target, and multi-pathway approach to disease treatment.

目的:探究治尪汤联合甲氨蝶呤改善胶原诱导关节炎模型(CIA)大鼠的骨破坏作用及机制.方法:SD雄性大鼠随机分为正常组,模型组,甲氨蝶呤组(1.5 mg·kg-1),甲氨蝶呤+治尪汤低(40.8 g·kg-1),高(102.0 g·kg-1)剂量组.除正常组外,其余大鼠分别于第1,7天尾部注射浓度为2 g·L-1的Ⅱ型胶原乳剂50μL建立CIA模型;第8天开始连续给药21d,每7d检测1次体质量、足趾厚度,进行关节炎指数(AI)评分,酶联免疫吸附测定法检测血清肿瘤坏死因子-α(TNF-α),白细胞介素-6(IL-6),白细胞介素-1β(IL-1β)含量,抗酒石酸酸性磷酸酶染色法检测踝关节破骨细胞的数量,免疫组化(IHC),实时荧光定量聚合酶链式反应(Real-time PCR),蛋白免疫印迹法(Western blot)检测踝关节核转录因子-κB受体活化因子配体(RANKL),骨保护素(OPG)的表达.结果:与正常组比较,模型组大鼠体质量显著降低(P＜0.01),足趾厚度显著增加(P＜0.01),踝关节AI评分显著增加(P＜0.01),血清TNF-α,IL-6,IL-1β含量显著升高(P＜0.01),踝关节破骨细胞数量显著增加(P＜0.01),踝关节RANKL mRNA和蛋白表达显著升高(P＜0.01),OPG mRNA和蛋白表达明显降低(P＜0.05),RANKL/OPG显著升高(P＜0.01).与模型组比较,甲氨蝶呤+治旭汤低、高剂量可改善大鼠体质量,降低足趾厚度,AI评分,RANKL mRNA和蛋白表达并增加OPG mRNA和蛋白表达,使RANKL/OPG降低(P＜0.05,P＜0.01),且甲氨蝶呤+治旭汤高剂量效果最好,甲氨蝶呤+治尪汤低剂量次之,甲氨蝶呤再次之.结论:治尪汤联合甲氨蝶呤能更好地改善类风湿关节炎的骨破坏,可能是通过抑制RANKL/OPG通路实现的,本研究为中西医结合方案治疗类风湿关节炎骨破坏提供了实验基础.