AIM To investigate the improvement effects of Poria cocos polysaccharides(PCPs)on mouse nonalcoholic fatty liver disease(NAFLD).METHODS Forty-eight C57BL/6 mice were randomly divided into the blank group,the model group,the simvastatin group(4 mg/kg)and the high,medium and low dose PCPs groups(200,100 and 50 mg/kg),with 8 mice in each group.The NAFLD model was reproduced by 16 weeks feeding of high-fat and high-cholesterol diet,followed by 8 weeks administration of corresponding drug by gavage.The mice had their body mass and liver coefficient assessed;their levels of hepatic free fatty acid(FFA),and serum total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),aspartate aminotransferase(AST),alanine aminotransferase(ALT),γ-glutamyltransferase(γ-GT)and malondialdehyde(MDA)detected;their hepatic pathological changes and lipid deposition observed using HE staining,NAFLD activity score(NAS)and oil red O staining;and their hepatic protein expressions of Akt,mTOR,p-Akt,p-mTOR and SREBP-1c detected by Western blot.RESULTS Compared with the blank group,the model group demonstrated all increased body weight,liver coefficient,hepatic FFA level,and serum TC,TG,LDL-C,AST,ALT,γ-GT,MDA,IL-1β and TNF-α.levels(P＜0.05,P＜0.01);decreased HDL-C level and activities of SOD and GSH-Px(P＜0.05,P＜0.01);more obvious hepatic pathological damage as revealed by increased NAS score(P＜0.01)and increased lipid deposition area(P＜0.01).Compared with the model group,the groups intervened with high or medium dose PCPs,or simvastatin displayed decreased body weight,liver coefficient,hepatic FFA level,and serum TC,TG,LDL-C,AST,ALT,γ-GT,MDA,IL-1β and TNF-α levels(P＜0.05,P＜0.01);increased HDL-C level and SOD,GSH-Px activities(P＜0.05,P＜0.01);decreased hepatic pathological damage as revealed by the decreased NAS score and lipid deposition area(P＜0.05,P＜0.01);and decreased hepatic protein expressions of p-Akt,p-mTOR and SREBP-1c protein(P＜0.05)as well.CONCLUSION PCPs can improve mouse NAFLD,and its mechanism may lie in their function in reversing abnormal lipid metabolism via Akt/mTOR/SREBP-1c signaling pathway.

Objective:To evaluate the impact of prophylactic cranial irradiation (PCI) on the prognosis of patients with limited-stage small cell lung cancer (SCLC) in the era of widespread application of MRI.Methods:Clinical data were collected from an open-lable prospective clinical trial on thoracic radiotherapy target volumes for limited-stage SCLC conducted in Sun Yat-sen University Cancer Center and Zhejiang Cancer Hospital between June 2002 and January 2017. In this study, patients who achieved complete remission (CR) or partial remission (PR) after definitive chemoradiotherapy (CRT) were retrospectively analyzed. Stratified analysis was performed according to different clinical efficacies. Patients were divided into different groups according to whether PCI was conducted or not. Survival analysis of patients was carried out. Survival data were calculated by Kaplan-Meier method, and Cox proportional hazards model was applied for multivariate prognostic analysis.Results:Among 309 patients with limited-stage SCLC who received CRT, 133 patients achieved CR and 140 cases obtained PR. These 273 patients were enrolled in this study. Among 133 patients with CR, 29 of them did not receive PCI, and 89 (85.6%) of the remaining 104 patients receiving PCI underwent brain MRI to exclude brain metastasis (BM) before PCI. With a median follow-up time of 22.1 months, the cumulative BM rates were 18.3% and 37.9% in patients who received or did not receive PCI ( P=0.020). The median overall survival (OS) was 30.2 and 30.5 months, and the 1-, 3- and 5-year OS rates were 93.3%, 41.9%, 27.7% and 82.8%, 44.8%, 40.8%, respectively ( P=0.910). Multivariate analysis indicated that baseline Karnofsky performance status (KPS) = 90 was a favorable independent prognostic factor for OS in CR patients ( HR=0.93, 95% CI: 0.89-0.98, P=0.006). Among 140 patients achieving PR, 52 cases did not receive PCI and 80 (90.9%) of the remaining 88 patients received brain MRI before PCI. With a median follow-up time of 18.9 months, the cumulative BM rates were 10.2% and 44.2% ( P<0.001). The median OS was 26.0 and 18.0 months, and the 1-, 3-, and 5-year OS rates were 86.4%, 37.9%, 32.2% and 75.0%, 17.3%, 10.8%, respectively ( P=0.001). Baseline KPS = 90 ( HR=0.93, 95% CI: 0.89-0.97, P=0.001) and PCI ( HR=0.54, 95% CI: 0.36-0.80, P=0.002) were favorable prognostic factors for OS in PR patients. Conclusions:PCI significantly reduces the incidence of BM and prolongs the OS in patients with limited-stage SCLC who achieve PR after CRT, but it fails to significantly prolong the OS of CR patients.

Objective:To investigate the ultra-long-term antihypertensive efficacy, safety, major adverse events, and survival benefits of renal denervation (RDN) in patients with resistant hypertension (rHTN) and mild chronic kidney disease (CKD).Methods:This real-world, single-center retrospective study enrolled patients with rHTN and mild CKD who underwent RDN at Tianjin First Central Hospital between October 2011 and June 2016. Office blood pressure, home self-measured blood pressure, 24-hour ambulatory blood pressure, serum creatinine, estimated glomerular filtration rate, and urine albumin-to-creatinine ratio were collected at baseline and at 1, 5, and 13 years post-RDN. The total daily defined dose of antihypertensive medications at 13 years post-RDN was recorded, along with endpoint events during follow-up, including cardiovascular death, all-cause death, hospitalization for heart failure, myocardial infarction, and stroke. Patients were stratified according to CKD stage (G1-G2 vs. G3a) and baseline systolic blood pressure (mild-to-moderate vs. severe hypertension), and follow-up data were compared across subgroups.Results:A total of 40 patients were included, aged (51±15) years, including 26 (65%) males. At the 13-year follow-up, office systolic blood pressure (SBP) and diastolic blood pressure (DBP) decreased by (-32±20) mmHg and (-15±14) mmHg (1 mmHg=0.133 kPa), respectively; reductions in home self-measured blood pressure (SBP: (-25±14) mmHg, DBP: (-10±11) mmHg) and 24-hour ambulatory blood pressure (SBP: (-16±9 mmHg, DBP: (-10±6) mmHg) were also observed, alongside a reduction in the total daily defined dose of antihypertensive medications by (1.1±0.9) compared to baseline. Renal function assessments showed no significant differences at 13 years versus baseline in serum creatinine ((105±51) μmol/L vs. (96±22) μmol/L), estimated glomerular filtration rate ((72±22) ml·min -1·1.73 m -2 vs. (78±17) ml·min -1·1.73 m -2), or urine albumin-to-creatinine ratio ((101±86) mg/g vs. (127±82) mg/g) (all P>0.05). All-cause and cardiovascular mortality rates during follow-up were 13% (5/40) and 8% (3/40), respectively. Subgroup analysis results showed that, although CKD G1-G2 patients had smaller reductions in office SBP ((-31±20) mmHg vs. (-34±19) mmHg) and DBP ((-13±10) mmHg vs. (-25±18) mmHg) compared to G3a patients at 13 years, intergroup differences were not significant (all P>0.05). In contrast, severe hypertension subgroup exhibited greater reductions in office SBP ((-55±13) mmHg vs. (-20±10) mmHg) and DBP ((-24±17) mmHg vs. (-13±10) mmHg) versus mild-to-moderate hypertension subgroup (all P<0.05). Conclusion:RDN demonstrates sustained antihypertensive efficacy with favorable renal safety in rHTN patients with mild CKD. Patients with higher baseline systolic blood pressure may exhibit better responsiveness to RDN.

AIM To investigate effects of petroleum ether fraction from Derris eriocarpa How on glucose and lipid metabolism in a mouse model of metabolic syndrome(MS).METHODS KM mice were fed a high-fat diet and administered streptozotocin intraperitoneally to establish MS models.The MS mice were then randomly assigned to the model group,the metformin hydrochloride group,the lovastatin group,the ursolic acid group,and the high-,medium-and low-dose D.eriocarpa petroleum ether fraction groups,with 10 mice in each group.Ten additional mice maitained on a normal diet served as the normal control group.After 4 weeks of intragastric administration,glucose and lipid metabolism indicators were measured.Hepatic pathological changes were assessed using HE staining and oil red O staining.Liver tissue mRNA expressions of ATF3,PEPCK,FXR,CYP7A1,HNF4ɑ,CYP8B1 and SRB1 were quantified by RT-qPCR.Hepatic protein expressions of ATF3,HNF4ɑ,PEPCK,FXR and CYP7A1 was analyzed by Western blot in MS mice.RESULTS Compared to the model group,the high-dose D.eriocarpa petroleum ether fraction group exhibited significant glucose tolerance improvement(reduced OGTT-AUC,P＜0.01);favorable serum lipid modulation in terms of increased HDL-C levels(P＜0.01)and decreased TG,TC,LDL-C(P＜0.01);reduced renal biomarkers(BUN,SCR)and hepatotoxic indicators of TBA,AST and ALT activities(P＜0.01);alleviated hepatic lipid accumulation and histopathological damage;downregulated mRNA and protein expressions of ATF3,HNF4ɑ and PEPCK,as well as CYP8B1 mRNA expression(P＜0.01);and upregulated mRNA and protein expressions of FXR and CYP7A1,along with SRB1 mRNA expression(P＜0.01).CONCLUSION D.eriocarpa petroleum ether fraction ameliorates glucose and lipid metabolism dysregulation in MS mice by modulating the ATF3/HNF4ɑ/CYP7A1 signaling pathway,consequently eliciting hypoglycemic,hypolipidemic,hepatoprotective and nephroprotective effects.

ObjectiveJunctophilin-2 (JPH2) is an essential structural protein that maintains junctional membrane complexes (JMCs) in cardiomyocytes by tethering the plasma membrane to the sarcoplasmic reticulum, thereby facilitating excitation-contraction (E-C) coupling. Mutations in JPH2 have been associated with hypertrophic cardiomyopathy (HCM), but the molecular mechanisms governing its membrane-binding properties and the functional relevance of its membrane occupation and recognition nexus (MORN) repeat motifs remain incompletely understood. This study aimed to elucidate the structural basis of JPH2 membrane association and its implications for HCM pathogenesis. MethodsA recombinant N-terminal fragment of mouse JPH2 (residues1-440), encompassing the MORN repeats and an adjacent helical region, was purified under near-physiological buffer conditions.X-ray crystallography was employed to determine the structure of the JPH2 MORN-Helix domain. Sequence conservation analysis across species and junctophilin isoforms was performed to assess the evolutionary conservation of key structural features. Functional membrane-binding assays were conducted using liposome co-sedimentation and cell-based localization studies in COS7 and HeLa cells. In addition, site-directed mutagenesis targeting positively charged residues and known HCM-associated mutations, including R347C, was used to evaluate their effects on membrane interaction and subcellular localization. ResultsThe crystal structure of the mouse JPH2 MORN-Helix domain was resolved at 2.6 Å, revealing a compact, elongated architecture consisting of multiple tandem MORN motifs arranged in a curved configuration, forming a continuous hydrophobic core stabilized by alternating aromatic residues. A C-terminal α-helix further reinforced structural integrity. Conservation analysis identified the inner groove of the MORN array as a highly conserved surface, suggesting its role as a protein-binding interface. A flexible linker segment enriched in positively charged residues, located adjacent to the MORN motifs, was found to mediate direct electrostatic interactions with negatively charged phospholipid membranes. Functional assays demonstrated that mutation of these basic residues impaired membrane association, while the HCM-linked R347C mutation completely abolished membrane localization in cellular assays, despite preserving the overall MORN-Helix fold in structural modeling. ConclusionThis study provides structural insight into the membrane-binding mechanism of the cardiomyocyte-specific protein JPH2, highlighting the dual roles of its MORN-Helix domain in membrane anchoring and protein interactions. The findings clarify the structural basis for membrane targeting via a positively charged linker and demonstrate that disruption of this interaction—such as that caused by the R347C mutation—likely contributes to HCM pathogenesis. These results not only enhance current understanding of JPH2 function in cardiac E-C coupling but also offer a structural framework for future investigations into the assembly and regulation of JMCs in both physiological and disease contexts.

The PA-PB1 interface of the influenza polymerase is an attractive site for antiviral drug design. In this study, we designed and synthesized a mini-library of indazole-containing compounds based on rational structure-based design to target the PB1-binding interface on PA. Biological evaluation of these compounds through a viral yield reduction assay revealed that compounds 27 and 31 both had a low micromolar range of the half maximal effective concentration (EC₅₀) values against A/WSN/33 (H1N1) (8.03 μmol/L for 27; 14.6 μmol/L for 31), while the most potent candidate 24 had an EC₅₀ value of 690 nM. Compound 24 was effective against different influenza strains including a pandemic H1N1 strain and an influenza B strain. Mechanistic studies confirmed that compound 24 bound PA with a K _d which equals to 1.88 μmol/L and disrupted the binding of PB1 to PA. The compound also decreased the lung viral titre in mice. In summary, we have identified a potent anti-influenza candidate with potency comparable to existing drugs and is effective against different viral strains. The therapeutic options for influenza infection have been limited by the occurrence of antiviral resistance, owing to the high mutation rate of viral proteins targeted by available drugs. To alleviate the public health burden of this issue, novel anti-influenza drugs are desired. In this study, we present our discovery of a novel class of indazole-containing compounds which exhibited favourable potency against both influenza A and B viruses. The EC₅₀ of the most potent compounds were within low micromolar to nanomolar concentrations. Furthermore, we show that the mouse lung viral titre decreased due to treatment with compound 24. Thus our findings identify promising candidates for further development of anti-influenza drugs suitable for clinical use.

The continuous emergence of SARS-CoV-2 variants as well as other potential future coronavirus has challenged the effectiveness of current COVID-19 vaccines. Therefore, there remains a need for alternative antivirals that target processes less susceptible to mutations, such as the formation of six-helix bundle (6-HB) during the viral fusion step of host cell entry. In this study, a novel high-throughput screening (HTS) assay employing a yeast-two-hybrid (Y2H) system was established to identify inhibitors of HR1/HR2 interaction. The compound IMB-9C, which achieved single-digit micromolar inhibition of SARS-CoV-2 and its Omicron variants with low cytotoxicity, was selected. IMB-9C effectively blocks the HR1/HR2 interaction in vitro and inhibits SARS-CoV-2-S-mediated cell-cell fusion. It binds to both HR1 and HR2 through non-covalent interaction and influences the secondary structure of HR1/HR2 complex. In addition, virtual docking and site-mutagenesis results suggest that amino acid residues A930, I931, K933, T941, and L945 are critical for IMB-9C binding to HR1. Collectively, in this study, we have developed a novel screening method for HR1/HR2 interaction inhibitors and identified IMB-9C as a potential antiviral small molecule against COVID-19 and its variants.

BACKGROUND: Pre-transplant exposure to immune checkpoint inhibitors (ICIs) significantly increases the risk of allograft rejection after liver transplantation (LT); however, whether ICI-related rejection leads to increased graft loss remains controversial. Therefore, this study aimed to investigate the association between ICI-related allograft rejection and perioperative graft loss. METHODS: This was a retrospective analysis of adult liver transplant recipients with early biopsy-proven T-cell-mediated rejection (TCMR) at Liver Transplantation Center of Sun Yat-sen Memorial Hospital from June 2019 to September 2024. The pathological features, clinical characteristics, and perioperative graft survival were analyzed. RESULTS: Twenty-eight patients who underwent early TCMR between June 2019 and September 2024 were included. Based on pre-LT ICI exposure, recipients were categorized into ICI-related TCMR (irTCMR, n = 12) and conventional TCMR (cTCMR, n = 16) groups. Recipients with irTCMR had a higher median Banff rejection activity index (RAI) (6 vs . 5, P = 0.012) and more aggressive tissue damage and inflammation. Recipients with irTCMR showed higher proportion of treatment resistance, achieving a complete resolution rate of only 8/12 compared to 16/16 for cTCMR. Graft loss occurred in 5/12 of irTCMR recipients within 90 days after LT, with no graft loss in cTCMRs recipients. Cox analysis demonstrated that irTCMR with an ICI washout period of <30 days was an independent risk factor for perioperative graft loss (hazard ratio [HR], 6.540; 95% confidence interval [CI], 1.067-40.067, P = 0.042). CONCLUSION IrTCMR is associated with severe pathological features, increased resistance to treatment, and higher graft loss in adult liver transplant recipients.
Humans ; Liver Transplantation/adverse effects* ; Male ; Female ; Middle Aged ; Retrospective Studies ; Graft Rejection/immunology* ; Immune Checkpoint Inhibitors/therapeutic use* ; Adult ; T-Lymphocytes/drug effects* ; Graft Survival/immunology* ; Aged