Premature ovarian insufficiency (POI) is characterized by complex etiologies and currently lacks effective treatments, posing significant threats to women’s physical and psychological health. Quercetin (QUE), a bioactive flavonoid inherent in traditional herbs such as Flos Sophorae and Cuscuta chinensis, demonstrates potent redox-modulating and anti-senescence capacities. This investigation validates its therapeutic efficacy against POI, specifically elucidating granulosa cell-centric mechanisms. POI mouse models and injury models of the KGN (human ovarian granulosa cell line) were induced through the administration of cyclophosphamide (CTX). Subsequently, these models were treated with drugs such as QUE. Morphological changes in the ovaries of the mice were observed, and vaginal smears were conducted to monitor the estrus cycles and patterns of the mice. ELISA was employed to detect sex hormone levels in various mouse groups, while Hematoxylin-eosin (H E) staining was utilized to assess the status of ovarian follicles at different stages. Moving forward, immunohistochemical analysis and Western blot were conducted to detect the expression levels of Sirtuin 1 (SIRT1), Hypoxia-Inducible Factor 1 Alpha (HIF-1α), and other apoptosis-related proteins. Ultimately, methods such as Terminal deoxynucleotidyl transferase dUTP Nick-End Labeling (TUNEL) assay, β-galactosidase staining, flow cytometry for apoptosis detection, and 5-Ethynyl-2′-deoxyuridine (EdU) assay were utilized to evaluate the apoptosis, senescence, and proliferation of mouse ovarian tissues and human ovarian granulosa cells KGN. QUE alleviated CTX-induced ovarian atrophy and follicular atresia by upregulating SIRT1 expression. Furthermore, QUE partially restored estrous cyclicity, normalized sex hormone levels, and improved follicular counts through SIRT1/HIF-1α modulation. Concurrently, QUE suppressed apoptosis in both murine and human granulosa cells via the SIRT1/HIF-1α pathway. Our findings demonstrate that QUE enhances ovarian reserve by inhibiting granulosa cells apoptosis via SIRT1/HIF-1α signaling, highlighting its therapeutic potential for POI management.

Background: Chaihu Erchen Decoction (CED) is a classical traditional Chinese medicine prescription widely used for disorders associated with phlegm accumulation and liver qi stagnation. However, the plasma-exposed constituents and metabolic characteristics responsible for its pharmacological activities remain largely unclear.Methods: An integrated metabolomic strategy was employed to characterize the chemical constituents, plasma-accessible compounds, and in vivo metabolites of CED. Comparative plasma profiling was performed in rats and mice to evaluate species-dependent differences in systemic exposure and metabolic disposition.Results: Extraction with 30% ethanol provided the highest extraction efficiency and was selected for subsequent investigation. A total of 200 constituents were characterized in CED, among which flavonoids represented the predominant chemical class. Following oral administration, 48 and 140 plasma-accessible constituents were identified in rat and mouse plasma, respectively, indicating markedly greater systemic exposure in mice. Furthermore, 96 putative metabolites were characterized. Flavonoids dominated both the circulating parent compounds and metabolite profiles. Extensive Phase I and Phase II biotransformation, including O-dealkylation, hydroxylation, oxidation, glucuronidation, and sulfation, was observed. Polymethoxylated flavonoids underwent characteristic sequential metabolic transformations and contributed substantially to the circulating metabolite pool.Conclusion: CED exhibited flavonoid-dominated systemic exposure and pronounced species-dependent metabolic characteristics. Extensive biotransformation of flavonoids, particularly polymethoxylated flavonoids, generated diverse circulating metabolites through coordinated Phase I and Phase II reactions.

To elucidate whether Danggui Sini Decoction (DGSD) relieves sciatica by downregulating transient receptor potential (TRP) channels involved in peripheral sensitization. After conducting chronic constriction injury of sciatic nerve (CCI) operation, 32 SD rats, randomly assigned to the sham, CCI 7d, CCI 14d, and CCI 21d groups (n=8 per group), were used to observe the course of sciatica. mRNA sequencing of dorsal root ganglia (DRG) was conducted on rats in the sham and CCI 21d groups. Another 32 rats were divided into the sham, CCI, mecobalamin, and DGSD groups, 8 in each group. mRNA sequencing analysis and single-cell RNA-sequencing (scRNA-seq) data analysis were conducted. Behavioural tests were conducted to evaluate the therapeutic effects of DGSD. The levels of TRP vanilloid (TRPV)1–4, TRP ankyrin 1 (TRPA1), TRP melastatin 8 (TRPM8), Ras homolog family member A (RhoA), and phosphorylated p38 mitogen-activated protein kinase (p-p38MAPK) were measured via Western blot, and the serum levels of C-C motif chemokine ligand 2 and 5 (CCL2, CCL5), substance P (SP), and prostaglandin E2 (PGE2) were conducted via ELISA. scRNA-seq and mRNA sequencing analysis revealed that inflammatory mediator regulation of TRP channels played a role in the development of sciatica. CCI rats exhibited cold hyperalgesia from day 4 to 21, increased the DRG protein expression levels of TRP channels, including TRPV1–4, TRPA1, TRPM8, RhoA, p-p38MAPK, CCL2, CCL5, SP, and PGE2. Treatment with DGSD decreased the serum levels of CCL2, CCL5, SP, and PGE2, and reduced the levels of RhoA, p-p38MAPK, TRPV1–4, TRPA1, and TRPM8 in DRG (P<0.05). CCI increased the levels of TRP channels and inflammatory factors, inducing cold hyperalgesia, whereas DGSD relieved pain via inhibiting peripheral sensitization to relieve cold hyperalgesia by reducing the inflammatory factors and TRP.

Imidacloprid (IMI), a widely used neonicotinoid insecticide, has been increasingly linked to metabolic disorders from environmental exposure. In this study, adult rats were chronically exposed to low-dose IMI at 0.06 mg/kg body weight per day for 19 weeks under three dietary regimens: normal-fat, high-fat, and free-choice diets. Prolonged IMI treatment significantly increased body weight, induced dyslipidemia characterized by elevated serum total cholesterol and triglycerides, reduced high-density lipoprotein cholesterol and triggered systemic inflammation. Notably, under normal-fat diet, IMI disrupted lipid metabolism via crosstalk between intestinal TAS2Rs (TAS2Rs) and fat receptor 41 (GPR41). Under high-fat and free-choice diets, the effects are mainly associated with cluster of differentiation 36 (CD36) mediated PKC-AMPK-Acetyl CoA carboxylase 1 (ACC1) signaling pathway. However, the relationship between this pathway and the decreased expression of TAS2Rs remains unclear, indicating that a high-fat dietary state may interfere with the flavor perception toxicity of IMI. The result of 16S rRNA sequencing showed IMI reduced gut microbial diversity, increased pathogenic Clostridium, and decreased beneficial Bacteroides. Spearman correlation analysis indicated that TAS2R119 expression positively correlated with Bacteroides abundance and negatively with norank_o__Clostridia_UCG-014, suggesting a link between receptor modulation and microbial community shifts. Molecular docking and gene expression analyses supported IMI interaction with TAS2Rs as a novel mechanism underlying pesticide-induced metabolic disruption. These findings illuminate the combined effects of diet and environmental chemicals on metabolism and identify intestinal taste receptors as potential therapeutic targets.

BackgroundSciatica causes severe pain and impaired mobility. Neuroinflammation is involved in the development of sciatica.PurposeThis study aimed to explore whether Gallic acid (GA) reduces neuroinflammation to relieve sciatica by regulating NOX4-mediated oxidative stress.MethodsAfter scRNA-seq analysis was performed, 32 SD (Sprague-Dawley) rats were randomly divided into 4 groups: sham operation, chronic constriction injury (CCI), CCI+mecobalamin, and CCI+GA groups. We conducted behavioral tests, ELISA, western blotting, and immunofluorescence analysis. In cell experiments, we conducted ROS measurement, flow cytometry, PCR, and western blotting.ResultsscRNA-seq analysis revealed that gene signatures related to the “inflammatory response” and “oxidative stress” were significantly enriched, with higher module scores observed specifically in M1 macrophages. In RAW264.7 cells, LPS stimulation significantly increased ROS generation and MDA levels and upregulated the expression of M1 macrophage markers, including IL-1β, iNOS, TNF-α, and CD32. In addition, LPS increased the protein expression of NOX4 and inflammatory mediators (TNF-α, IBA-1, IL-1β, COX-2, and iNOS) while reducing the levels of ATF4 and p-Nrf2. GA treatment reduced ROS generation and MDA levels; downregulated the mRNA expression of IL-1β, iNOS, TNF-α, and CD32; and increased CD206 mRNA expression. Similarly, GA decreased the protein levels of NOX4, TNF-α, IBA-1, IL-1β, COX-2, and iNOS but restored the expression of ATF4 and p-Nrf2. In CCI rats, GA significantly attenuated thermal hyperalgesia from Day 7 to Day 21, with thermal withdrawal thresholds recovering toward sham control levels. CCI markedly increased IL-8, COX-2, TNF-α, TGF-β, IL-6, and IL-1β levels in the sciatic nerve; increased IBA-1/CD32 coexpression; decreased IBA-1/CD206 coexpression; and markedly disrupted sciatic nerve architecture. These pathological changes were accompanied by elevated expression of IBA-1, NOX4, IL-1β, and iNOS, together with reduced ATF4 and p-Nrf2 levels. Notably, GA treatment largely reversed these CCI-induced alterations.ConclusionGA alleviated sciatica in a rat model, possibly through its ability to promote the polarization of proinflammatory M1 macrophages toward anti-inflammatory M2 macrophages via the regulation of NOX4-mediated oxidative stress.

The aim of this research endeavor was to explore the therapeutic potential of ( +)-catechin in mitigating neuropathic pain. A total of thirty-two Sprague‒Dawley rats were randomly allocated into four groups: the sham group, the chronic constriction injury (CCI) group, the CCI + ibuprofen group, and the CCI + ( +)-catechin group. The results of the in vivo experiment show that ( +)-catechin has the potential to improve mechanical hyperalgesia induced by CCI and reduce the infiltration of inflammatory cells in the injured sciatic nerve. CCI induces the upregulation of nNOS, iNOS, IL-1β, and COX-2 within the rat sciatic nerve and leads to an elevation in the levels of IL-1β, PGE2, and TNF-α in the serum of rats, while simultaneously diminishing the secretion of IL-10. Moreover, immunofluorescence analysis reveals that CCI enhances the expression of CD32 (an M1 polarization marker) in the rat spinal cord, while diminishing the expression of CD206 (an M2 polarization marker). However, the administration of ( +)-catechin effectively counteracts these effects. Western blot analysis further demonstrates that ( +)-catechin significantly reduces the protein expression of IBA-1, IL-1β, MyD88, p-NF-κB, p-JNK, p-ERK, p-p38MAPK, COX-2, and TLR4 within the spinal cord. The findings of the BV2 cell experiment revealed the attenuating effects of ( +)-catechin on M1 polarization markers (such as IL-1β, TNF-α, iNOS, and CD32), while concurrently boosting the levels of M2 polarization markers (including CD206, IL-10, and Arg-1). Notably, administration of LPS significantly heightened the accumulation of IBA-1, IL-1β, MyD88, p-NF-κB, p-JNK, p-ERK, p-p38MAPK, TLR4, COX-2, and iNOS, while concurrently suppressing Arg-1 expression. However, the administration of ( +)-catechin effectively reversed these alterations. Overall, these findings suggest that ( +)-catechin alleviates neuropathic pain by modulating the M1 and M2 phenotypes of microglia through the TLR4/MyD88/NF-κB pathway.

This study investigated the mechanism underlying Paeonol’s therapeutic efficacy against neuropathic pain. GSE158892 dataset data were used to conduct a scRNA-seq analysis. In cell experiments, Schwann cells and macrophages were utilized to examine pain pathogenesis using specific inhibitors. Thirty-two SD rats were randomly divided into four groups: sham, chronic constriction injury (CCI), ibuprofen, and Paeonol. Behavioral tests combined with ELISA, PCR, western blot, immunohistochemistry, and immunofluorescence analyses were conducted. CellChat analysis demonstrated that, following peripheral nerve injury, Schwann cells secreted IL-34, which interacted with CSF1R on macrophages, leading to the infiltration and activation of macrophages. Paeonol reduced IL-34 production by Schwann cells induced with LPS. Conditioned medium from LPS-stimulated Schwann cells treated with Paeonol did not cause macrophage proliferation or migration, activation of the CSF1 pathway, or ROS production. In CCI rats, Paeonol alleviated mechanical and cold hyperalgesia, while reducing the production of serum inflammatory mediators. Additionally, Paeonol decreased the expression levels of IL-34, CSF1R, phosphorylated ERK (p-ERK), phosphorylated NF-κB (p-NF-κB), and components of the NLRP3 inflammasome in the dorsal root ganglia of CCI rats. Conclusion: Alleviation of neuropathic pain by Paeonol treatment may be achieved by inhibiting the IL-34–CSF1R interaction, suppressing Schwann cell–macrophage interactions, and reducing DRG neuroinflammation.

INTRODUCTION:Sciatica causes intense pain. No satisfactory therapeutic drugs exist to treat sciatica. This study aimed to probe the potential mechanism of ferulic acid in sciatica treatment.METHODS:Thirty-two SD rats were randomly divided into 4 groups: sham operation, chronic constriction injury (CCI), mecobalamin, and ferulic acid. We conducted RNA sequencing, behavioral tests, ELISA, PCR, western blotting, and immunofluorescence analysis. TAK-242 and JSH23 were administered to RSC96 and GMI-R1 cells to explore whether ferulic acid can inhibit apoptosis and alleviate inflammation.RESULTS:RNA sequencing showed that TLR4/NF-κB pathway is involved in the mechanism of sciatica. CCI induced cold and mechanical hyperalgesia; destroyed the sciatic nerve structure; increased IL-1β, IL-6, TNF-α, IL-8, and TGF-β protein levels and IL-1β, IL-6, TNF-α, TGF-β, TLR4, and IBA-1 mRNA levels; and decreased IL-10 and INF-γ protein levels and IL-4 mRNA levels. Immunohistochemistry showed that IBA-1, CD32, IL-1β, iNOS, nNOS, COX2, and TLR4 expression was increased while S100β and Arg-1 decreased. CCI increased TLR4, IBA-1, IL-1β, iNOS, Myd88, p-NF-κB, and p-p38MAPK protein levels. Treatment with mecobalamin and ferulic acid reversed these trends. Lipopolysaccharide (LPS) induced RSC96 cell apoptosis by reducing Bcl-2 and Bcl-xl protein and mRNA levels and increasing Bax and Bad mRNA and IL-1β, TLR4, Myd88, p-NF-κB, and p-p38MAPK protein levels, while ferulic acid inhibited cell apoptosis by decreasing IL-1β, TLR4, Myd88, p-NF-κB, and p-p38MAPK levels and increasing Bcl-2 and Bcl-xl levels. In GMI-R1 cells, Ferulic acid attenuated LPS-induced M1 polarization by decreasing the M1 polarization markers IL-1β, IL-6, iNOS, and CD32 and increasing the M2 polarization markers CD206, IL-4, IL-10 and Arg-1. After LPS treatment, IL-1β, iNOS, TLR4, Myd88, p-p38MAPK, and p-NF-κB levels were obviously increased, and Arg-1 expression was reduced, while ferulic acid reversed these changes.CONCLUSION:Ferulic acid can promote injured sciatic nerve repair by reducing neuronal cell apoptosis and inflammatory infiltration though the TLR4/NF-κB pathway.

对《伤寒论》条文中涉及的单纯数脉主病、数脉相兼脉主病和数脉相类脉主病等内容进行整理与分析.《伤寒论》中的数脉所反映出来的病机纷繁复杂,脉义具有多重性.单纯数脉主要反映虚证、提示病情变化;数脉相兼脉主要反映虚证、里实热证、秘结的大便与宿食等有形之邪、正邪斗争的疾病态势、痈疡病等;数脉相类脉主要反映里实热证、虚证、正邪斗争胶着的疾病态势.根据数脉主病可辨明病势是整体向愈还是整体恶化;脉证合参更有利于病机的判别.数脉涉及病机的核心在于"热"与"阳",可以是阴虚所导致的虚热,或是正虚所导致的阳不秘(阳之失调),或是实邪导致的阳盛.

通过《伤寒论》自身条文自证、前后条文互证的方法,同时旁引历代著名医家关于《伤寒论》的条文注解,如《注解伤寒论》《伤寒贯珠集》《医宗金鉴》等相关原文,从医古文文理及临床医理两个方面,对《伤寒论》条文第331条、176条进行解读.认为《伤寒论》第331条现行句读方式有误,而第176条的现行解释"脉浮滑,为表里俱热"引用第350条"伤寒脉滑而厥者,里有热,白虎汤主之"进行理解更有说服力.

Therapeutic drugs of chronic neuralgia have a high risk of addiction, making it crucial to identify novel drugs for chronic neuralgia. This study aimed to explore the therapeutic effect of paeoniflorin on chronic sciatica via inhibiting Schwann cell apoptosis. 28 SD rats were randomly divided into four groups, including the sham operation group, chronic constriction injury (CCI) group, mecobalamin group, and paeoniflorin group. The therapeutic effect and mechanism of paeoniflorin were evaluated via rat and cell experiments. Mechanical, hot, or cold hyperalgesia was induced in the rats after CCI operation, while paeoniflorin relieved chronic neuralgia. Besides, paeoniflorin decreased the levels of IL1, IL6, TNF‐α, CRP, and LPS and increased the level of IL10 in serum. As for the sciatic nerve, the number of inflammatory cells was decreased, and Schwann cells were present after paeoniflorin treatment, and paeoniflorin promoted the recovery of nerve structure. In cell experiments, LPS induced Schwann cell apoptosis via the TLR4/NF‐kB pathway. And paeoniflorin attenuated LPS‐induced Schwann cell apoptosis by decreasing the levels of TLR4, p‐NF‐kB, caspase3, cleaved‐caspase3, and cleaved‐caspase7. Overall, these results suggest that paeoniflorin alleviates chronic sciatica by decreasing inflammatory factor levels and promotes the repair of damaged nerves by reducing Schwann cell apoptosis.

Oxidative stress damage is known as one of the important factors that induce neuropathic pain (NP). Using antioxidant therapy usually achieves an obvious curative effect and alleviates NP. Previous pharmacological studies have shown that higenamine (Hig) performs to be antioxidant and anti-inflammatory. However, the protective effect and mechanism of Hig on NP are still unclear. This study mainly evaluated the changes in reactive oxygen species (ROS) level, lipid peroxidation, and antioxidant system composed of superoxide dismutase (SOD) and glutathione (GSH) through chronic constrict injury (CCI) model rats and t-BHP-induced Schwann cell (SC) oxidative stress model. The expressions of two inflammatory factors, tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), were also assessed. The possible molecular mechanism of Hig in the treatment of NP was explored in conjunction with the expression of mitochondrial apoptosis pathway and NOX2/ROS/TRP/P38 mitogen-activated protein kinase (MAPK)/NF-ĸB pathway-related indicators. Hig showed substantial antioxidant and anti-inflammatory properties both in vivo and in vitro. Hig significantly reduced the upregulated levels of ROS, malondialdehyde (MDA), TNF-α, and IL-6 and increased the levels of SOD and GSH, which rebalanced the redox system and improved the survival rate of cells. In the animal behavioral test, it was also observed that Hig relieved the CCI-induced pain, indicating that Hig had a pain relief effect. Our research results suggested that Hig improved NP-induced oxidative stress injury, inflammation, and apoptosis, and this neuroprotective effect may be related to the NOX2/ROS/TRP/P38 MAPK/NF-ĸB signaling pathway.

Purpose. We explored the role of ROS in cold-induced vasoconstriction and corresponding mechanism. Methods. Three experiments were performed. First, we measured blood flow in human hands before and after cold exposure. Second, 24 mice were randomly divided into 3 groups: 8 mice received saline injection, 8 received subcutaneous Tempol injection, and 8 received intrathecal Tempol injection. After 30 min, we determined blood flow in the skin before and after cold exposure. Finally, we used Tempol, CCG-1423, and Go 6983 to pretreat HAVSMCs and HUVECs for 24 h. Then, cells in the corresponding groups were exposed to cold (6 h, 4°C). After cold exposure, the cytoskeleton was stained. Intracellular Ca2+ and ROS levels were measured by flow cytometry and fluorescence microscopy. We measured protein expression via Western blotting. Results. In the first experiment, after cold exposure, maximum skin blood flow decreased to 118.4 ± 50.97 flux units. Then, Tempol or normal saline pretreatment did not change skin blood flow. Unlike intrathecal Tempol injection, subcutaneous Tempol injection increased skin blood flow after cold exposure. Finally, cold exposure for 6 h shrank the cells, making them narrower, and increased intracellular Ca2+ and ROS levels in HUVECs and HAVSMCs. Tempol reduced cell shrinkage and decreased intracellular Ca2+ levels. In addition, Tempol decreased intracellular ROS levels. Cold exposure increased RhoA, Rock1, p-MLC-2, ET-1, iNOS, and p-PKC expression and decreased eNOS expression. Tempol or CCG-1423 pretreatment decreased RhoA, Rock1, and p-MLC-2 levels in HAVSMCs. Furthermore, Tempol or Go 6983 pretreatment decreased ET-1, iNOS, and p-PKC expression and increased eNOS expression in HUVECs. Conclusion. ROS mediate the vasoconstrictor response within the cold-induced vascular response, and ROS in blood vessel tissues rather than nerve fibers are involved in vasoconstriction via the ROS/RhoA/ROCK1 and ROS/PKC/ET-1 pathways in VSMCs and endothelial cells.

中医临床基础学科主要课程包括《伤寒论》《金匮要略》及《温病学》3门课程.此文从教师层面、学生层面对中医临床基础课程融合教学的作用进行分别论述,认为融合教学体现了中医经典理论从源到流的传承和发展过程,学术思想、学术观点一脉相承的特点,对提高中医类本科生中医临床基础课程的教学效果,对学生中医临床思维的培养和中医临床诊治能力的提升有着不可忽视的重要作用.

This study was to investigate the protective effect of paeoniflorin (PF) on hydrogen peroxide-induced injury. Firstly, "SMILES" of PF was searched in Pubchem and further was used for reverse molecular docking in Swiss Target Prediction database to obtain potential targets. Injury-related molecules were obtained from GeenCards database, and the predicted targets of PF for injury treatment were selected by Wayne diagram. For mechanism analysis, the protein-protein interactions were constructed by String, and the KEGG analysis was conducted in Webgestalt. Then, cell viability and cytotoxicity assay were established by CCK8 assay. Also, the experimental cells were allocated to control, model (200 μmol·L-1 H2O2), SB203580 10 μmol·L-1 (200 μmol·L-1 H2O2+ SB203580 10 μmol·L-1), PF 50 μmol·L-1 (200 μmol·L-1 H2O2+ PF 50 μmol·L-1), and PF 100 μmol·L-1 (200 μmol·L-1 H2O2+ PF 100 μmol·L-1) groups. We measured the intracellular ROS, Hoechst 33258 staining, cell apoptosis, the levels of Bcl-xl, Bcl-2, Caspase-3, Cleaved-caspase3, Cleaved-caspase7, TRPA1, TRPV1, and the phosphorylation expression of p38MAPK. There are 96 potential targets that may be associated with PF for injury treatment. Then, we chose the "Inflammatory mediator regulation of TRP channels" pathway for the experimental verification from the first 10 KEGG pathway. In experimental verification, H2O2 decreased the cell viability moderately (P < 0.05), and 100 μmol·L -1 PF increased the cell viability significantly (P < 0.05). Depending on the difference of intracellular ROS fluorescence intensity, PF inhibited H 2O2-induced reactive oxygen species production in Schwann cells. In Hoechst 33258 staining, PF reversed the condensed chromatin and apoptotic nuclei following H2O2 treatment. Moreover, Flow cytometry results showed that PF could substantially inhibit H2O2 induced apoptosis (P < 0.05). Pretreatment with PF obviously reduced the levels of Caspase3, Cleaved-caspase3, Cleaved-caspase7, TRPA1, TRPV1, and the phosphorylation expression of p38MAPK after H 2O2 treatment (P < 0.05), increased the levels of Bcl-2, and Bcl-xl ( P < 0.05). PF inhibited Schwann cell injury and apoptosis induced by hydrogen peroxide, which mechanism was linked to the inhibition of phosphorylation of p38MAPK.