Abstract: Objective To explore the mechanism of Bushen Wenyang Huayu formula in regulating the Notch1 / Twist1 signaling pathway-mediated epithelial-mesenchymal transition ( EMT ) in the treatment of endometriosis. Methods SD female rats were randomly divided into Sham operation group, Model group, BWH-H group, BWH-L group, and Dienogest group. The EMs rat model of Kidney Yang Deficiency was constructed by autologous transplantation combined with ice water bath. After the modeling was completed, BWH-H group and BWH-L group were respectively given high and low-dose medicinal solutions of Bushen Wenyang Huayu formula by gavage. Dienoges group was given dienogest tablets by gavage, while Sham operation group and Model group were given an equal volume of distilled water by gavage for 21 consecutive days, weight, volume, and adhesion degree of ectopic lesions and uteri were observed. Serum levels of sex hormones were determined by enzyme-linked immunosorbent assay, E-Cadherin, N-Cadherin, Vimentin, Notch1, and Twist1 proteins were detected by immunohistochemistry and Western blot, and gene expression levels of E-Cadherin, N-Cadherin, Vimentin, Notch1,and Twist1 were detected by RT-qPCR. Results Serum levels of estradiol ( E2), progesterone ( P ), folliclestimulating hormone (FSH), and luteinizing hormone (LH) were significantly increased in Model group compared with Sham operation group (P<0. 05). N-Cadherin, Vimentin, Notch1, and Twist1 protein and mRNA levels in uterine tissue were also significantly increased (P<0. 01, P<0. 05), while E-Cadherin protein and mRNA levels were significantly decreased (P<0. 01). The volume and weigh of ectopic lesions was significantly reduced in BWH-H group and BWH-L group traditional Chinese medicine and Dienogest group, compared with Model group (P<0. 05,P<0. 01), while E2, P, FSH, and LH levels were significantly decreased ( P<0. 05), N-Cadherin, Vimentin,Notch1, and Twist1 protein and mRNA levels in the uterine and ectopic lesion tissues were significantly decreased(P<0. 01, P<0. 05 ), and E-Cadherin protein and mRNA levels were significantly increased ( P< 0.01 ). Conclusions Bushen Wenyang Huayu formula can significantly inhibit the progression of endometriosis, and its mechanism may be related to the down-regulation of the Notch1/ Twist1 signaling pathway, thereby regulating the EMT process.

Abstract: Objective To explore the mechanism by which “Three Methods and Three Acupoints” massage improves motor function in sciatic nerve injury (SNI) rats by delaying denervated muscle atrophy. Methods Thirtysix male Sprague-Dawley rats were divided randomly into normal, sham operation, model, and massage groups (n=9). Rats in model and massage groups were subjected to SNI model establishment by the clamping method, and rats in the sham operation group underwent sciatic nerve exposure without clamping. Interventions were carried out in rats in the massage group from the 7th day after surgery. Point, pull, and knead techniques were applied to the Yinmen,Chengshan, and Yanglingquan acupoints in sequence, using an intelligent massage technique simulation instrument.Each acupoint was treated with each technique for 1 minute, once a day, with 1 day rest after 10 interventions,followed by another 10 day intervention. Rats in sham operation and model groups received the same duration of grasping and fixation treatment, and rats in normal group maintained routine feeding conditions. The hind limb grip force was measured before modeling and after 10 and 20 interventions to evaluate muscle strength. After 20 massage sessions, the rats were sacrificed and the right gastrocnemius muscle was removed for hematoxylin-eosin staining to observe muscle atrophy and measure the cross-sectional area of the muscle fibers. mRNA levels of CCNB1, CNTF,and MEF2A in the right gastrocnemius muscle tissue were detected by reverse transcription-polymerase chain reaction,paired box (PAX) 3, Desmin, hepatocyte growth factor (HGF) protein expression levels in the right gastrocnemius muscle were detected by Western blot, and MEF2C protein expression in the affected gastrocnemius muscle was detected by immunofluorescence. Results The hind limb grip force was significantly reduced in model rats compared with normal and sham operation groups (P<0. 01). The muscle fibers were also disordered in model rats, the muscle cells were significantly atrophied, and the cross-sectional area was significantly reduced ( P<0. 01), and mRNA expression levels of CCNB1, CNTF, and MEF2A, and protein levels of PAX3, Desmin, HGF, and MEF2C were lower compared with the normal and sham operation groups (P<0. 05). The hind limb grip force was significantly increased in the massage group compared with the model group (P<0. 01), the muscle fibers were atrophied but arranged in an orderly manner, and the cross-sectional area was significantly increased ( P<0. 01), and CCNB1,CNTF, and MEF2A mRNA and PAX3, Desmin, HGF, and MEF2C protein levels were significantly upregulated (P<0. 05). Conclusions “Three Methods and Three Acupoints” can promote the gene expression of CCNB1, CNTF,and MEF2A and protein levels of PAX3, Desmin, HGF, and MEF2C, promote the proliferation and differentiation of muscle satellite cells, repair nerve injury, delay denervated muscle atrophy, and improve the motor function in SNI rats.

Abstract: Objective The pathological mechanism of inflammatory bowel disease (IBD) is closely related to overactivation of the nuclear factor-κB ( NF-κB ) signaling pathway and M1 polarization of macrophages.Accordingly, this study aimed to investigate the effects of Kangfuxin solution on the regulatory mechanism of the NF-κB pathway during the progression of IBD. Methods A mouse model of IBD was established using dextran sodium sulfate (DSS). Forty-eight C57BL / 6 mice were divided into eight groups: Control, Model, SASP, KFX-L (5 mL / kg), KFX-M (10 mL / kg), KFX-H (20 mL / kg), NF-κBi-pretreated and NF-κBi-pretreated+KFX-H groups. The disease activity index (DAI), colonic mucosal damage index (CMDI), and histopathological scores were recorded. mRNA expression levels of CD86, tumor necrosis factor-α (TNF-α), phosphorylated NF-κB inhibitor α (p-IκBα),and interleukin ( IL)-1β in colonic tissues were detected by RT-qPCR, and protein expression levels of CD86,inducible nitric oxide synthase, and p-IκBα in colon tissues were detected by Western blot. Flow cytometry was used to analyze the effect of Kangfuxin liquid on the disease model. Results DSS-induction successfully established a mouse IBD model, with the measured values of various research indicators being highest in the Model group ( P<0. 05). The indicators in each group, except the SASP group, were significantly decreased compared with the Model group (P<0. 05), while downregulation of CD86 in the SASP group was not significant (P>0. 05). Among the three Kangfuxin liquid groups, the indicators were significantly reduced in the KFX-H group (P<0. 05). The combined intervention of NF-κB inhibitor and high-dose Kangfuxin liquid did not significantly enhance the therapeutic effect,with no significant differences in the measured values of each indicator compared with the single-drug groups ( P>0. 05). Conclusions Kangfuxin liquid, sulfasalazine and NF-κB inhibitors all have protective effects on colonic inflammation in DSS-induced IBD model mice, but their sites of action differ. Kangfuxin liquid and NF-κB inhibitors synergistically attenuate the intestinal inflammatory response by inhibiting NF-κB pathway activation through reducing IκBα phosphorylation, as well as down-regulating macrophage M1-type polarization. In contrast, sulfasalazine exerts a weaker effect on macrophage M1 polarization.

Abstract: Objective To elucidate the role of G2 / S phase expressed 1 (GTSE1) in carboplatin resistance mechanisms in ovarian cancer. Methods (1) GTSE1 protein expression was compared between carboplatin-sensitive (OVCAR3, OV3R-CBP, COC1) cell lines by Western blot. (2) GTSE1 was then silenced in OV3R-CBP cells using small interfering ( si) RNA transfection si-GTSE1 or si-normal control (NC), and the knockdown efficiency was confirmed by quantitative reverse transcription-polymerase chain reaction and Western blot. Carboplatin dose-response assays (0, 10, 20, 40, 80 and 160 μmol / L) were conducted to determine the half-maximal inhibitory concentration (IC50 ) and drug resistance index. (3) OV3R-CBP cells were treated with various combinations: carboplatin alone (70 μmol / L, CBP group), carboplatin plus LY2109761 ( 10 μmol / L, CBP +si+LY2109261 group), carboplatin plus si-NC (CBP +si-NC group), carboplatin plus si-GTSE1(CBP +si-GTSE1 group), and a triple combination of carboplatin, si-GTSE1 and XMU-MP-1 ( 1 μmol / L,CBP +si-GTSE1+XMU-MP-1 group). Expression levels of Ecadherin, Vimentin, Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ) were detected by Western blot. Cell proliferation inhibition and apoptosis were assessed by Cell Counting Kit-8 assay and flow cytometry. Results (1) GTSE1 expression was significantly elevated in OV3R-CBP relative to OVCAR3 /COC1 cells (P<0. 05). ( 2 ) Knockdown of GTSE1 significantly decreased GTSE1 mRNA/ protein levels and carboplatin IC50 and resistance index (P<0. 05). ( 3) Silencing GTSE1 in the presence of carboplatin suppressed YAP, TAZ, and Vimentin expression, and enhanced E-cadherin expression, YAP / TAZ phosphorylation, cell proliferation inhibition, and apoptosis (P<0. 05). Hippo pathway inhibition using XMU-MP-1 reversed these effects,indicating pathway-specific modulation. Conclusions Silencing GTSE1 reverses carboplatin resistance in ovarian cancer cells by activating the Hippo-YAP / TAZ pathway and inhibiting epithelial-mesenchymal transition. Targeting GTSE1 restores chemosensitivity via reactivation of the Hippo pathway, offering a potential therapeutic strategy for overcoming platinum resistance in ovarian cancer.

Abstract: Objective To explore the dose-time-dependent relationship of shear stress and different exposure times on the expression of THBS4 in HASMCs, and to reveal its potential mechanism in vascular remodeling, with the aim of guiding the treatment of clinical cardiovascular diseases. Methods HASMCs were subjected to shear stress of 0, 4, 8 and 12 dyn / cm2 using a parallel plate flow chamber for 6 and 12 h, respectively, and the cells were collected after treatment. mRNA expression of THBS4 was detected by real-time fluorescence quantitative polymerase chain reaction and protein expression was detected by Western blot. The co-localization of THBS4 and cleaved-Caspase8 was examined by immunofluorescence staining. Results The mRNA and protein expression levels of THBS4 gradually increased with increasing shear stress and exposure duration ( P< 0. 05). THBS4 gene and protein levels were significantly higher in shear-H group than in control, shear-L and shear-M groups(P<0. 05). Immunofluorescence detection demonstrated that THBS4 was co-localized with cleaved-Caspase8 and exhibited consistent expression levels. Conclusions Spatiotemporal gradient changes in shear stress can regulate the expression of THBS4 in HASMCs, and its expression level is positively correlated with the magnitude of shear stress, with a time-dependent cumulative effect. These result suggest that THBS4 may modulate apoptotic pathways in HASMCs under shear stress.

Abstract: Objective To explore the effect of Xingnao Kaiqiao acupuncture on depressive behavior in poststroke depressed rats by adjusting the P2X7 receptor (P2X7R) / NOD-like receptor family pyrin domain containing 3 (NLRP3) / interleukin-1β (IL-1β) pathway. Methods A post-stroke depression rat model was constructed and rats were divided into model, positive control (fluoxetine), acupuncture, pathway inhibitor (CBBG), and acupuncture+ pathway activator ( acupuncture + BzATP ) groups. Healthy rats were included as control group. Depression-like behavior was detected by sugar preference test and open field test. Serum inflammatory and oxidative stress factors were measured by enzyme-linked immunosorbent assay. Pathological changes in hippocampal tissue were detected by hematoxylin-eosin and Nissl body staining. Neuronal apoptosis in hippocampal tissue was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling, and P2X7R, NLRP3, IL-1β, ionized calciumbinding adapter molecule 1 (Iba-1), B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), and cleaved caspase3 proteins in hippocampal tissue were detected by Western blot. Results Compared with control group, rats in model group showed decreased glucose preference and open field activity distance, pathological damage to brain tissue, increased serum inflammatory factors, malondialdehyde ( MDA), and proportion of neuronal apoptosis, decreased catalase ( CAT) activity, increased levels of P2X7R/ NLRP3 / IL-1β pathway proteins, Iba-1, and pro apoptotic proteins in hippocampal tissue, and decreased anti-apoptotic proteins (P<0. 05). Compared with model group, rats in the model, fluoxetine, acupuncture, and CBBG groups showed increased glucose preference and open field activity distance, reduced pathological damage in hippocampal tissue, increased number of Nissl bodies, decreased levels of serum inflammatory factors, MDA, and proportion of neuronal apoptosis, increased CAT activity, decreased levels of P2X7R/ NLRP3 / IL-1β pathway proteins, Iba-1, and pro apoptotic proteins in hippocampal tissue, and increased antiapoptotic proteins (P<0. 05). Compared with acupuncture group, rats in the acupuncture + BzATP group showed aggravated depressive behavior, clear pathological damage to hippocampal tissue, and a decrease in the number of Nissl bodies, as well as inflammation and oxidative stress reactions, increased levels of P2X7R/ NLRP3 / IL-1β pathway proteins, Iba-1, and pro-apoptotic proteins, and decreased levels of anti-apoptotic proteins decreased (P<0. 05). Conclusions Xingnao Kaiqiao acupuncture may alleviate inflammation and the oxidative stress response and inhibit neuronal apoptosis in rats by regulating the P2X7R/ NLRP3 / IL-1β pathway, thereby improving hippocampal tissue damage and exerting antidepressant effects.

Abstract: Objective To establish a multiple special staining method combined with image processing protocols tailored for bone tissue with varying degrees of decalcification. Methods Fifteen 10-week-old SpragueDawley rats were selected and humanely euthanized. The right femurs were harvested and fixed, followed by acid decalcification at different intensities. Paraffin-embedded sections were prepared and stained with hematoxylin-eosin to evaluate the degree of decalcification. The sections were then stained with toluidine blue, safranin O-fast green, and a multiple special staining protocol, and the staining efficacies of each method in terms of bone morphology under varying decalcification conditions were compared. Spectral unmixing techniques were also employed to develop an image processing strategy specifically for multiple-stained samples. Results Toluidine blue and safranin O-fast green staining showed suboptimal result in over-decalcified samples, while the optimized multiple special staining protocol provided enhanced visualization of cortical bone, chondrocytes, and collagen structures in over-decalcified tissues.Spectral processing resolved composite images into individual dye-specific components, thereby improving the accuracy of the quantitative analysis. Conclusions The established multiple special staining method significantly improves the visualization of cortical bone, cartilage, and collagen in over-decalcified samples. This technique offers a valuable tool for morphological studies related to bone metabolic diseases.

Abstract: Objective To address limitations of conventional behavioral experimental apparatuse and to design an intelligent, integrated, and precise novel stimulation device for psychology experiments to investigate the facilitative effects of adrenoceptor agonists on learned anxiety. Methods We developed an integrated behavioral experiment stimulation device combining electrical / acoustic stimulation, visual observation, thermostatic control, and computerized operation. The core components included a control system, main microprocessor, power supply, electrical stimulator, noise stimulator, thermostatic unit, and visual observation chamber. We used the device to deliver conditioned auditory stimuli paired with unconditioned foot-shock stimuli to induce anxiety in rats. Results The device eliminated the drawbacks of fragmented operations and achieved precise synchronization of electrical and noise stimuli, ensured strict correlation between conditioned and unconditioned stimuli, and enabled programmable preset stimulation sequences. The device accordingly reduced the risk of operational error, significantly improved experimental repeatability, and increased students’ scientific understanding of anxiety behaviors. The device also demonstrated applicability to pain research and other behavioral experiments. Conclusions As a multifunctional instrument, this novel device represents a valuable addition to the traditional medical curriculum, by addressing teaching-oriented demands through medicine-engineering integration, advancing course development, and enhancing talent cultivation.

Abstract:Summarize and analyze the reporting of outcome indicators in current clinical randomized controlled trials (RCTs) on acupuncture intervention for gastrointestinal dysfunction in sepsis, summarize the types of outcome indicators, and put forward appropriate suggestions to promote the progress of clinical research. Four Chinese and three English databases and one clinical trial registry were systematically searched to screen out RCTs of needleassisted treatment for gastrointestinal dysfunction in sepsis. The risk of bias was assessed using the Cochrane risk of bias tool. After screening, a total of 30 clinical studies were finally retrieved, including 28 Chinese and two English studies. Thirty-nine outcome indicators were extracted and mainly summarized into eight categories: Traditional Chinese Medicine syndromes, clinical effective rate, gastrointestinal function, disease severity, imaging examinations, serological markers, inflammatory indicators, and safety. Most studies had problems such as unclear risk of bias, poor quality of research method ologies, confusion of primary and secondary outcome measures, nonstandard use of outcome measures, and lack of acupuncture characteristics. We therefore collected and summarized the core outcome indicators of acupuncture treatment for gastrointestinal dysfunction in sepsis, clarified the limitations of the outcome indicators of acupuncture treatment for gastrointestinal dysfunction in sepsis, and identified research deficiencies to improve the method ological quality of clinical research.

Abstract:Liver depression syndrome is commonly observed in various clinical diseases. Extensive studies have demonstrated that chronic stress serves as a primary trigger for liver depression syndrome, and its biological mechanisms are closely related to changes in the morphology and function of multiple central brain regions. The relevant evidence, however, remains scattered across numerous studies, necessitating further synthesis and review. This review accordingly summarizes the relationships between the pathogenesis of liver depression syndrome and morphological,functional, and molecular changes in brain regions, including the hippocampus, cortex, hypothalamus, locus coeruleus, and amygdala. Among these, the pathological changes in liver depression syndrome models are predominantly concentrated in the hippocampus and hypothalamus. However, distinct liver depression syndrome models exhibit variations in the affected brain regions. This review summarizes and explores the common biological alterations and specific differences among different liver depression syndrome models. The findings provide a foundation and direction for further research into the biological basis of liver depression syndrome and the identification of syndrome-specific targets.

Abstract:Sepsis-induced myocardial injury ( SIMI) is a common and potentially fatal complication of sepsis. Its pathogenesis has not been fully elucidated, but involves multiple pathological processes, including excessive inflammatory response, mitochondrial dysfunction, apoptosis, and oxidative stress. Current clinical interventions primarily include fluid resuscitation, vasoactive agents, and anti-inflammatory therapies; however, their efficacy remains limited, and they fail to reverse pathological remodeling at the epigenetic level. This systematic review summarizes the result of studies on epigenetic modifications, including DNA methylation, histone modifications (acetylation, methylation), and non-coding RNA regulation, in relation to septic myocardial injury. The review aims to provide novel perspectives to aid in the early warning, diagnosis, treatment, and targeted drug development of SIMI, thereby improving the prognosis of patients with this condition.

Abstract:Programmed cell death is currently a critical focus in cancer therapy research. Among its various forms, ferroptosis and disulfidptosis have recently attracted considerable attention as novel mechanisms of cell death.Ferroptosis is an iron-dependent, peroxide-driven form of cell death, whereas disulfidptosis is induced by disulfide stress result ing from cystine accumulation during glucose deprivation. Both types of cell death are closely associated with tumor progression, and recent studies have demonstrated that their activation may serve as an effective strategy for cancer intervention. SLC7A11 plays a dual regulatory role in both ferroptosis and disulfidptosis, by inhibiting ferroptosis under normoglycemic conditions, while promoting disulfidptosis in glucose-deprived environments.Targeting the expression and metabolic reprogramming role of SLC7A11 and developing synergistic strategies to induce both ferroptosis and disulfidptosis may thus offer novel theoretical and clinical approaches for cancer therapy.

Abstract:Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by chronic synovitis and pannus formation, with an unknown etiology. The Wnt / β-catenin signaling pathway has emerged as a critical therapeutic target for RA. Despite recent research progress, systematic reviews and integrated analyses remain inadequate. This review explores the mechanisms underlying the role of the Wnt / β-catenin signaling pathway in the pathological progression of RA, including its regulation of inflammatory responses, fibroblast-like synoviocyte proliferation, apoptosis, bone destruction, and angiogenesis pathways. It also summarizes the latest research advancements in the modulation of the Wnt / β-catenin signaling pathway by traditional Chinese medicine monomers (such as phenolics, terpenoids, and quinones) and compound formulations (including Chinese patent medicines and other compound types) for the treatment of RA, with the aim of providing theoretical foundations and reference directions for future clinical applications and the development of novel drugs for RA.

Abstract:Metabolic dysfunction-associated steatotic liver disease (MASLD) is a globally prevalent chronic liver disease. As a complex condition caused by multiple factors, the precise pathogenesis of MASLD remains incompletely understood; however, inflammation plays a central role in the pathological development of MASLD.NOD-like receptor family pyrin domain containing 3 (NLRP3) is a critical immune regulatory factor with a pivotal role in the inflammatory response of MASLD. Overactivation of NLRP3 exacerbates chronic liver inflammation and promotes fat accumulation in the liver. Furthermore, excessive activation of NLRP3 leads to the generation of reactive oxygen species, which further activates immune response cells, thereby intensifying liver damage. Inhibiting NLRP3 inflammasome activation has thus become a novel strategy for treating MASLD. Numerous studies have indicated that extracts and formulations of traditional Chinese medicine can effectively alleviate the inflammatory response in MASLD by inhibiting NLRP3 activation, achieving significant progress in its clinical treatment. This review considers the role of the NLRP3 signaling pathway in MASLD and the potential of traditional Chinese medicine for improving MASLD by regulating NLRP3 inflammasome activity, providing a reference for the prevention and treatment of MASLD with traditional Chinese medicine.