This study aimed to investigate the protective effects of Huangqi Guizhi Wuwu decoction (HGWD) against cerebral ischemic injury and the underlying mechanisms. A middle cerebral artery occlusion (MCAO) model was established in C57BL/6 mice to evaluate the effects of HGWD on neurobehavioral scores, cerebral infarction rate, brain water content, and oxidative stress and inflammatory markers. The mRNA and protein expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax) in brain tissue were assessed. In addition, Nrf2 knockout mice were used to verify the role of Nrf2 in the protective effects of HGWD against MCAO-induced injury. Additionally, an oxygen-glucose deprivation/reperfusion (OGD/R) model in primary neuronal cells was employed to further confirm the pharmacological effects of HGWD in vitro. The results showed that HGWD significantly ameliorated cerebral ischemic injury in MCAO mice, alleviated oxidative stress, suppressed the release of inflammatory factors, and markedly upregulated the expression of Nrf2, HO-1, and Bcl-2 while downregulating Bax expression, with consistent trends being observed at both mRNA and protein levels. The protective effects of HGWD were significantly attenuated in Nrf2 knockout mice, indicating the pivotal role of Nrf2 in HGWD-mediated protection against cerebral ischemic injury. In vitro experiments revealed that HGWD significantly increased neuronal cell viability, reduced lactate dehydrogenase(LDH) leakage, and decreased apoptosis in OGD/R-treated cells, accompanied by upregulation of Nrf2, HO-1, and Bcl-2 and downregulation of Bax. In conclusion, HGWD protects against cerebral ischemic injury by activating the Nrf2/HO-1 pathway to enhance antioxidant capacity and modulating the Bcl-2/Bax signaling pathway to inhibit apoptosis, thereby protecting brain cells from ischemic damage.

ObjectiveTo investigate the mechanism of action of Jiedu Huayu granules in improving liver injury in mice with acute liver failure （ALF） by observing its effect on a mouse model of ALF after prophylactic administration， and to provide a basis for clinical medication. MethodsA total of 60 specific pathogen-free male C57BL/6J mice were divided into normal group， model group， Jiedu Huayu granules group （JDHY group）， and farnesoid X receptor （FXR） agonist （GW4064） group using a random number table， with 15 mice in each group. The model of ALF was induced by a single intraperitoneal injection of D-galactosamine combined with lipopolysaccharide. The mice in the JDHY group were given prophylactic administration of 0.3 g/mL drug solution of Jiedu Huayu granules by gavage for 3 days before modeling， those in the normal group and the model group were given 0.9% NaCl solution by gavage， and those in the GW4064 group were given intraperitoneal injection of GW4064 for 3 consecutive days before modeling. The mice were sacrificed after modeling， and serum and liver tissue samples were collected. A veterinary automatic biochemical analyzer was used to measure the serum levels of total bilirubin （TBil）， total bile acids （TBA）， gamma-glutamyl transferase （GGT）， alanine aminotransferase （ALT）， and aspartate aminotransferase （AST） in mice from each group； HE staining was used to observe liver pathological changes； RT-PCR was used to measure the mRNA expression levels of FXR， fibroblast growth factor 15 （FGF15）， fibroblast growth factor receptor 4 （FGFR4）， small heterodimer partner （SHP）， and bile salt export pump （BSEP） in mice， and Western blot was used to measure the protein expression levels of FXR， FGF15， FGFR4， SHP， and BSEP. A one-way analysis of variance was used for comparison between groups， and the Dunett method was used for further comparison between two groups. ResultsCompared with the normal group， the model group had significant increases in the serum levels of TBil， ALT， AST， TBA， and GGT （all P<0.01）， and compared with the model group， the JDHY group and the GW4064 group had significant reductions in the serum levels of TBil， ALT， AST， TBA， and GGT （all P <0.01）. HE staining showed that compared with the model group， the JDHY group and the GW4064 group had milder pathological injury， a reduction in the area of hepatocyte necrosis， and alleviation of cellular swelling and edema. Compared with the normal group， the model group had significant reductions in the mRNA and protein expression levels of FXR， FGF15， FGFR4， SHP， and BSEP in liver tissue （all P <0.01）， and compared with the model group， the JDHY group and the GW4064 group had significant increases in the mRNA and protein expression levels of FXR， FGF15， FGFR4， SHP， and BSEP in liver tissue （all P <0.05）. ConclusionJiedu Huayu granules may alleviate liver injury in mice with ALF through the FXR/SHP axis.

Objective To investigate the effects and mechanisms of high-fat diet(HFD)-induced obesity on male ze-brafish reproductive function.Methods Adult male zebrafish were divided into normal diet(ND)group and HFD group.Growth and metabolic conditions were evaluated by measuring body weight,body length,BMI,organ index,and glucose/lipid lev-els.Reproductive capacity was assessed via sperm concentration,motility,fertilization rate,and testosterone levels.Testicular tissues from both of groups were subjected to transeriptomic sequencing(RNA-seq).And qRT-PCR was used to validate the expression of genes.Results Compared to male zebrafish in ND group,the ones in HFD group exhibited hepatic steatosis and glucose/lipid meta-bolic disorders(P＜0.05).Testicular structural disorganization,along with reduced testosterone levels,decreased gonadosomatic in-dex,and impaired sperm concentration and motility occurred in HFD group(P＜0.05).GO analysis revealed that differentially ex-pressed genes were enriched in spermatogenesis and ciliary system,while KEGG analysis highlighted metabolic related pathways(pu-rine metabolism,thyroid hormone synthesis,mTOR signaling)and cell adhesion molecules.Twenty key differentially expressed genes were validated by qRT-PCR,which confirmed the reliability of RNA-Seq results.Conclusion Impairment of reproductive function induced by HFD in zebrafish may be associated with three regulatory mechanisms including ciliary system,metabolic dysregulation,and aberrant cell adhesion molecule signaling.This study provides mechanistic insights and identifies potential therapeutic targets for clinical management of diet-associated infertility.

ObjectiveTo establish steroid-induced osteonecrosis of the femoral head （SANFH） cell model by using dexamethasone （DEX）-induced bone marrow mesenchymal stem cells （BMSCs） and demonstrate that Gushing Granules （GSNs） exert an improving effect by activating the phosphatidylinositol-3-kinase/protein kinase B/B-lymphoma-2 gene related promoter （PI3K/Akt/Bad） signalling pathway. MethodsFirstly， SD rats were orally administered with drugs at a dose of 0.9 g·kg^-1 to prepare GSN-containing serum， and CCK-8 screening was used to determine the optimal dosage and duration of action. Then， BMSCs were cultured and treated with 1×10^-6 mol·L^-1 DEX， 10% GSN-containing serum， and inhibitor LY294002 of PI3K/Akt signalling pathway for 24 hours to model and group SANFH cells. Cell viability and proliferation were detected by using CCK-8 assay kit and EdU staining kit. Flow cytometry was used to detect cell apoptosis. An alkaline phosphatase （ALP） assay kit was employed to detect ALP expression. In order to detect the PI3K/Akt/Bad signalling pathway and protein and mRNA expression of apoptosis-related proteins such as apoptosis regulatory factors B-cell lymphoma-2 gene （Bcl-2）， and Bcl-2-associated X protein （Bax）， osteocalcin （OCN）， and Collagen Ⅰ， we used Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultsThe CCK-8 assay kit determined that the optimal dosage for GSN-containing serum is 10%， and the duration of action is 48 hours. After modelling and grouping the cells in each group， the detection results showed that the SANFH model group had significantly lower cell viability， cell proliferation， and ALP expression， as well as protein and mRNA expressions of PI3K， Akt， Bad， Bcl-2， OCN， and Collagen I compared to the blank group. The nucleic acid and protein levels of the Bax index and the cell apoptosis rate detected by flow cytometry significantly increased （P<0.05，P<0.01）. After treatment with GSN-containing serum， cell viability， cell proliferation， and ALP expression， as well as expressions of PI3K， Akt， Bad， Bcl-2， OCN， and Collagen Ⅰ nucleic acids and proteins were significantly increased， while the nucleic acid and protein levels of the Bax index and the cell apoptosis rate detected by flow cytometry significantly decreased（P<0.05，P<0.01）. Compared with the GSN drug-containing serum group， the simultaneous treatment with the inhibitor LY294002 and GSN drug-containing serum reversed the improvement effect of GSN. Specifically， the cell viability， cell proliferation， ALP expression， and the nucleic acid and protein levels of PI3K， Akt， Bad， Bcl-2， OCN， and Collagen Ⅰ were all significantly decreased， while the nucleic acid and protein levels of the Bax index and the cell apoptosis rate detected by flow cytometry were significantly increased （P<0.05， P<0.01）. ConclusionGSNs antagonize DEX-induced apoptosis of BMSCs by activating the PI3K/Akt/Bad signalling pathway， providing a scientific theoretical basis for the clinical treatment of SANFH with GSNs.

Traditional Chinese Medicine (TCM), as a treasure of the Chinese nation, plays a significant role in maintaining public health. In 2019, the Central Committee of the Communist Party of China and the State Council proposed for the first time the establishment of a TCM registration and evaluation evidence system that integrates TCM theory, "personal experience" and clinical trials (referred to as the "Three-in-One" System) to promote the inheritance and innovation of TCM. Subsequently, the National Medical Products Administration issued several guiding principles to advance the improvement and implementation of this system. Owing to the complexity of its implementation, there are still differing understandings within the TCM industry regarding the positioning of the "Three-in-One" Registration and Evaluation Evidence System, as well as the connotation and value orientation of the "personal experience." To address this, Academician WANG Qi, President of the TCM Association, China International Exchange and Promotion Association for Medical and Healthcare and TCM master, led a group of academicians, TCM masters, TCM pharmacology experts and clinical TCM experts to convene a "Seminar on Promoting the Implementation of the ′Three-in-One′ Registration and Evaluation Evidence System for Chinese Medicinals." Through extensive discussions, an expert consensus was formed, clarifying the different roles of the TCM theory, "personal experience" and clinical trials within the system. It was further emphasized that the "personal experience" is the core of this system, and its data should be derived from clinical practice scenarios. In the future, the improvement of this system will require collaborative efforts across multiple fields to promote the high-quality development of the Chinese medicinal industry.

Objective:To study the mechanistic role of Jiedu Huayu granule in improving intestinal mucosal barrier functional damage and protecting against liver failure in mice induced by D-galactosamine combined with lipopolysaccharide.Methods:Fifty mice were randomly divided into a normal group, a model group, a low-dose Jiedu Huayu granule group (4.16 g·kg -1·d -1), a medium-dose Jiedu Huayu granule group (8.32 g·kg -1·d -1), and a high-dose Jiedu Huayu granule group (16.64 g·kg -1·d -1). D-galactosamine combined with lipopolysaccharide was administered once to establish an acute liver failure mouse model, followed by corresponding drug intervention. Retro-orbital blood samples were collected to measure serum alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels six hours after modeling in mice. The endotoxin content was detected using the limulus reagent method. The content of serum interleukin-1β, tumor necrosis factor-α, and interferon-γ levels were detected using enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was used to observe liver and intestinal pathological tissues.Western blot and RT-PCR were used to determine the protein and mRNA expression of Occludin, Zo-1, Tricellulin, and CLDN3 in intestinal tissues. Multiple quantitative data were analyzed using one-way ANOVA, following a normal distribution. LSD-t test was used for statistical analysis between inter-groups. Results:The medium-and high-dose Jiedu Huayu granules had reduced and improved liver function impairment, liver pathological tissue damage, intestinal barrier damage, serum endotoxin and inflammatory factors ( P<0.05), and increased intestinal expression of occludin, Zo-1, Tricellulin, and CLDN3 ( P<0.05) in mice group with acute liver failure. Conclusion:Jiedu Huayu granules can reduce liver inflammatory damage, gut-derived endotoxemia, bacterial endotoxins entering the liver, and protect intestinal barrier function, thereby remissioning liver inflammation and antagonizing liver failure in mice with acute liver failure.

Depression,as a severe psychological and psychiatric disorder,significantly impairs the long-term physical and mental health of patients.Current depression detection methods are plagued by strong subjectivity,limited techniques,and inadequate intelligence.Previous studies have mostly relied on single-modal signal analysis,making it difficult to comprehensively reflect the multidimensional characteristics of depression.Based on the independently developed intelligent depression detection system,wearable devices are used to collect prefrontal dual-lead EEG signals,PPG signals,and single-lead ECG signals.Data from 30 patients with depression and 40 healthy controls are collected and analyzed.A multimodal depression recognition model named RBLF-Net is proposed,which integrates spatiotemporal features,weighted attention,and random forests to utilize the multi-signal features for depression recognition.The model exhibits superior performance in the five-fold cross-validation,achieving a classification accuracy of 81.43%,a precision of 81.02%,and a recall rate of 81.25%,outperforming other comparative models,and thus providing an intelligent analysis approach for depression recognition from the perspective of multi-modal fusion.

Objective To investigate the predictive value of serum low-density lipoprotein cholesterol(LDL-C)and high-sensitive C-reactive protein(hs-CRP)in combination with carotid ultrasound parameters for assessing the risk of coronary heart disease(CHD)in patients with suspected cases.Methods A total of 260 patients who underwent coronary angiography(CAG)in the Department of Cardiology at Wujin Affiliated Hospital of Nanjing University of Traditional Chinese Medicine from August 2022 to August 2024 due to suspected CHD were enrolled in this study.Based on CAG results,patients were categorized into a non-CHD group(n=110)and a CHD group(n=150).General clinical characteristics,serum LDL-C,hs-CRP levels,and carotid ultrasound arameters-including carotid intima-media thickness(CIMT)and Crouse score of carotid artery plaque-were compared between the two groups.Spearman correlation analysis and multivariate logistic regression analysis were conducted to evaluate the associations among these parameters.The predictive value of individual and combined markers for suspected CHD was assessed by using receiver operating characteristic(ROC)curve analysis.Results(1)The coronary heart disease(CHD)group had higher age,smoking,fasting plasma glucose(FPG),and blood pressure compared to the non-CHD group(P>0.05).No significant difference was observed in the gender ratio,alcohol consumption rates,and body mass index(BMI),between the two groups(P>0.05).(2)Total cholesterol(TC),LDL-C,hs-CRP levels,and carotid ultrasound parameters were significantly higher in the CHD group than those in the non-CHD group(P<0.05).(3)Spearman correlation analysis and multivariate logistic regression analysis revealed that serum LDL-C,hs-CRP,and carotid ultrasound parameters were significantly correlated with coronary heart disease(P<0.05).(4)ROC curve analysis demonstrated that the combined assessment of serum LDL-C,hs-CRP,and carotid ultrasound parameters achieved the highest area under the curve(AUC),sensitivity,and specificity,with statistically significant differences compared to individual markers(P<0.05).Conclusions Serum LDL-C,hs-CRP,and carotid ultrasound parameters individually have predictive value for coronary heart disease,while their combined assessment offers superior predictive accuracy.The enhanced diagnostic performance of the combined valuation provides valuable guidance for the diagnosis and management of patients with suspected coro-nary heart disease.

This study aims to establish an efficient protoplast-mediated genetic transformation sys-tem for Pochonia chlamydosporia(P.chlamydosporia)to facilitate gene transformation,genetic modification,and subsequent biological function research.Through orthogonal testing,the study systematically optimized key factors influencing electroporation,including voltage,osmotic stabi-lizer,pulse time,nucleic acid concentration,and protoplast concentration.The results showed that the optimal electroporation conditions were:voltage of 250 V,osmotic stabilizer as 0.6 mol/L su-crose solution,pulse time of 10 ms,plasmid concentration of 1％,and protoplast concentration of 1×107 protoplasts/mL,yielding a transformation efficiency of 0.4 × 103 CFU/pg.The interaction analysis revealed that the five factors affected transformation efficiency in the following order:plasmid concentration＞protoplast concentration＞pulse time＞osmotic stabilizer type＞voltage.Based on the positive transformants,the study further evaluated their colony morphology,growth rate,conidial yield,mycelial dry weight,and the ability to infect the eggs of three types of animal gastrointestinal nematodes.The results indicated that the electroporation process did not signifi-cantly affect the biological functions of P.chlamydosporia.The transformants successfully ex-pressed enhanced green fluorescent protein(EGFP),and PCR analysis confirmed the successful in-tegration of the thiostrepton resistance gene.In conclusion,this study successfully established a protoplast-based electroporation-mediated genetic transformation system for P.chlamydosporia,providing a valuable foundation for further research into the mechanism of egg parasitism and the underlying genes involved in its biocontrol activity.

Objective To investigate the predictive value of serum low-density lipoprotein cholesterol(LDL-C)and high-sensitive C-reactive protein(hs-CRP)in combination with carotid ultrasound parameters for assessing the risk of coronary heart disease(CHD)in patients with suspected cases.Methods A total of 260 patients who underwent coronary angiography(CAG)in the Department of Cardiology at Wujin Affiliated Hospital of Nanjing University of Traditional Chinese Medicine from August 2022 to August 2024 due to suspected CHD were enrolled in this study.Based on CAG results,patients were categorized into a non-CHD group(n=110)and a CHD group(n=150).General clinical characteristics,serum LDL-C,hs-CRP levels,and carotid ultrasound arameters-including carotid intima-media thickness(CIMT)and Crouse score of carotid artery plaque-were compared between the two groups.Spearman correlation analysis and multivariate logistic regression analysis were conducted to evaluate the associations among these parameters.The predictive value of individual and combined markers for suspected CHD was assessed by using receiver operating characteristic(ROC)curve analysis.Results(1)The coronary heart disease(CHD)group had higher age,smoking,fasting plasma glucose(FPG),and blood pressure compared to the non-CHD group(P>0.05).No significant difference was observed in the gender ratio,alcohol consumption rates,and body mass index(BMI),between the two groups(P>0.05).(2)Total cholesterol(TC),LDL-C,hs-CRP levels,and carotid ultrasound parameters were significantly higher in the CHD group than those in the non-CHD group(P<0.05).(3)Spearman correlation analysis and multivariate logistic regression analysis revealed that serum LDL-C,hs-CRP,and carotid ultrasound parameters were significantly correlated with coronary heart disease(P<0.05).(4)ROC curve analysis demonstrated that the combined assessment of serum LDL-C,hs-CRP,and carotid ultrasound parameters achieved the highest area under the curve(AUC),sensitivity,and specificity,with statistically significant differences compared to individual markers(P<0.05).Conclusions Serum LDL-C,hs-CRP,and carotid ultrasound parameters individually have predictive value for coronary heart disease,while their combined assessment offers superior predictive accuracy.The enhanced diagnostic performance of the combined valuation provides valuable guidance for the diagnosis and management of patients with suspected coro-nary heart disease.