Extensive evidence has demonstrated that glucagon-like peptide-1 receptor agonists (GLP-1RAs) can ameliorate obesity. Our previous studies revealed that (Ex-4)₂-Fc, a long-acting GLP-1RA we developed, depends on the leptin pathway to treat obesity. However, the mechanisms linking (Ex-4)₂-Fc and leptin resistance remain largely unclear. To address this question, we explored the mechanism of GLP-1RAs from the perspective of the gut microbiota, as increasing evidence indicates an important link between the gut microbiota and obesity. This study aimed to explore the potential role of the gut microbiota in the treatment of GLP-1RAs. We found that (Ex-4)₂-Fc treatment reshaped obesity-induced gut microbiota disturbances and substantially increased the abundance of Akkermansia muciniphila (Am). In addition, (Ex-4)₂-Fc did not respond well in antibiotic-treated (ATB) Obese mice. Subsequent studies have shown that this defect can be overcome by gavage with Am. In addition, we found that Am enhanced (Ex-4)₂-Fc therapy by producing the metabolite inosine. Inosine regulates the macrophage adenosine A2A receptor (A2A) pathway to indirectly reduce leptin levels in adipocytes Thus, elucidating the role of metabolites in regulating the leptin pathway will provide new insights into GLP-1RAs therapy and may lead to more effective strategies for guiding the clinical use of antidiabetic agents.

Hypoxia-inducible factor (HIF-1α), a core transcription factor responding to changes in cellular oxygen levels, is closely associated with a wide range of physiological and pathological conditions. However, its differential impacts on vascular cell types and molecular programs modulating human vascular homeostasis and regeneration remain largely elusive. Here, we applied CRISPR/Cas9-mediated gene editing of human embryonic stem cells and directed differentiation to generate HIF-1α-deficient human vascular cells including vascular endothelial cells, vascular smooth muscle cells, and mesenchymal stem cells (MSCs), as a platform for discovering cell type-specific hypoxia-induced response mechanisms. Through comparative molecular profiling across cell types under normoxic and hypoxic conditions, we provide insight into the indispensable role of HIF-1α in the promotion of ischemic vascular regeneration. We found human MSCs to be the vascular cell type most susceptible to HIF-1α deficiency, and that transcriptional inactivation of ANKZF1, an effector of HIF-1α, impaired pro-angiogenic processes. Altogether, our findings deepen the understanding of HIF-1α in human angiogenesis and support further explorations of novel therapeutic strategies of vascular regeneration against ischemic damage.
Humans ; Vascular Endothelial Growth Factor A/metabolism* ; Endothelial Cells/metabolism* ; Transcription Factors/metabolism* ; Gene Expression Regulation ; Hypoxia/metabolism* ; Cell Hypoxia/physiology*

Food toxicology plays a crucial role in supporting scientific and technical aspects of food safety risk assessment.However,traditional methods relying on animal testing are becoming increas-ingly inadequate for identifying and evaluating emerging foods and unknown risks.There is a strong push worldwide towards the development of new approach methodologies(NAMs)based on non-animal testing methods.Policies and regulations related to NAMs are being standardized gradually in the European Union,the United States,and China.Some progress has also been made in applying these methodologies in food toxicology research in China.For instance,within the"Food Toxicology Program"at the National Center for Food Safety Risk Assessment,high-content and high-throughput in vitro hazard identification models employing model organisms like human macrophages,hepatocytes,adipocytes,embryonic stem cells,and zebrafish,as well as Toxicological Thresholds of Concern and quantitative in vitro to in vivo extrapolation based on physiologically-based toxicokinetic models have been estab-lished and applied.Nonetheless,new toxicological hazard identification technologies still face challenges such as inadequate elucidation of toxic mechanisms,insufficient collaborative research efforts,and inef-ficient translation of these findings into practical applications.

Tinosporae Radix， as a traditional Chinese medicinal herb， is the dried root tuber of Tinospora sagittata or T. capillipes. It was first recorded in the Compendium of Materia Medica Supplement in the Qing Dynasty and included in the previous edition of the Chinese Pharmacopoeia. Tinosporae Radix is excavated in autumn and winter and used after removing fibrous roots， washing， and drying. It is indicated for sore throat， carbuncle boils poison， waist and abdominal pain， and various heat syndromes and is commonly used to treat chronic inflammation. Its efficacy is significantly known as “broad-spectrum antibiotics in Zhuang medicine”. Tinosporae Radix is a traditional Chinese medicinal herb often taken by Zhuang and Yao nationalities in Guangxi province and has a wide range of application and development values and research significance. Modern studies have shown that Tinosporae Radix contains diterpenoids， alkaloids， sterols， anthraquinones， glycosides， fatty acids， volatile oils， and other compounds， which have many pharmacological activities such as anti-inflammatory and analgesic， antibacterial and antibacterial， antioxidant， anti-diabetic， and anti-tumor and anti-cancer effects， and it has achieved good efficacy in inhibiting inflammation and treating sore throat and other diseases. In recent years， there have been many research reports on the status， chemical constituents， pharmacological action， clinical application， and quality evaluation of Tinosporae Radix resources， but there is no systematic review and introduction at present. By consulting the literature and combining it with modern research， this paper systematically summarizes and collates Tinosporae Radix resources to provide guidance for the comprehensive development and utilization of Tinosporae Radix resources and subsequent in-depth study.

Age-dependent loss of skeletal muscle mass and function is a feature of sarcopenia, and increases the risk of many aging-related metabolic diseases. Here, we report phenotypic and single-nucleus transcriptomic analyses of non-human primate skeletal muscle aging. A higher transcriptional fluctuation was observed in myonuclei relative to other interstitial cell types, indicating a higher susceptibility of skeletal muscle fiber to aging. We found a downregulation of FOXO3 in aged primate skeletal muscle, and identified FOXO3 as a hub transcription factor maintaining skeletal muscle homeostasis. Through the establishment of a complementary experimental pipeline based on a human pluripotent stem cell-derived myotube model, we revealed that silence of FOXO3 accelerates human myotube senescence, whereas genetic activation of endogenous FOXO3 alleviates human myotube aging. Altogether, based on a combination of monkey skeletal muscle and human myotube aging research models, we unraveled the pivotal role of the FOXO3 in safeguarding primate skeletal muscle from aging, providing a comprehensive resource for the development of clinical diagnosis and targeted therapeutic interventions against human skeletal muscle aging and the onset of sarcopenia along with aging-related disorders.
Animals ; Humans ; Sarcopenia/metabolism* ; Forkhead Box Protein O3/metabolism* ; Muscle, Skeletal/metabolism* ; Aging/metabolism* ; Primates/metabolism*

Objective Myocardial functional outcome after revascularization of coronary chronic total occlusion (CTO) was prospectively predicted using extracellular volume fraction (ECV) based on cardiovascular magnetic resonance (CMR). Methods Thirty patients with CTO underwent CMR before and 6 months after percutaneous coronary intervention (PCI) were enrolled. The CMR scan protocol included cine, pre?contrast and post?contrast T1 mapping and late gadolinium enhancement (LGE). Ejection fraction (EF) and segmental wall thickening (SWT) were calculated using CVI 42 software. SWT less than 45% indicated myocardial segment dysfunction. According to the American Heart Association (AHA) scientific statement, the dysfunctional segments assigned to CTO vessel were selected, and three baseline imaging markers, ECV, transmural extent of infarction (TEI) and unenhanced rim thickness (RIM) were respectively evaluated. The myocardial segments were divided into two subgroups, group with well?developed collaterals and group with poorly?developed collaterals, based on the collateral circulation using Rentrop classification. Baseline and follow?up values of SWT and EF were evaluated using paired Student′s t?test. Using an increase in SWT>10% as standard reference, ROC analysis was conducted to describe the predictive performance of baseline markers. A mixed linear model was used to probe the relationship between collateral circulation and SWT. Stepwise logistic regression analysis was used to determine the independent predictors of regional functional recovery. The differences of EF between poorly?developed and well?developed collaterals were compared by Student t test. Results The baseline mean segmental wall thickening (SWT) of the dysfunctional segments increased from 21.6% (9.7%, 33.3%) to 38.4% (19.0%, 51.2%) after PCI (Z=-6.869, P<0.001), and EF was also significantly higher compared with baseline (54.5%±8.5 % vs. 50.7%± 6.6%, t=-5.706, P<0.001). ECV showed good performance in predicting functional recovery with cutoff value 34.7%, area under ROC curve (AUC) 0.86, sensitivity 91%, and specificity 66%. The AUC of ECV was superior to TEI and RIM (AUC: 0.75 and 0.73, all P value<0.01). The segments with well?developed collaterals were associated with a higher SWT at follow?up [46.6% (36.6%, 64.2%) vs. 33.5% (12.8%, 47.8%),F=5.791, P=0.02]. Logistic regression analysis demonstrated that mean segmental ECV was the only independent predictors of regional functional outcome after PCI (OR=0.83, 95% confidence interval: 0.77—0.89; P<0.001). Conclusions ECV by CMR may provide incremental value for the prediction of regional functional recovery in CTO patients, and baseline collateral circulation correlates with the regional systolic function after revascularization.

Objective: To study the airway inflammation feature difference in the patients with occupational pulmonary thesaurosis induced by iron. Methods: Induced sputum samples were obtained from 20 healthy men, 20 patients with stable asthma, 25 patients with occupational pulmonary thesaurosis induced by iron. Differential cell counts in induced sputum were examined. The levels of supernatant IL-8 were measured by ELISA. Results: The pulmonary function result was normal in the patients with pulmonary thesaurosis., their FVC (4.1±1.2) L was similar to FVC (4.3±0.9) L in mal group; but their FEV_1.0 (2.9±0.9) L、FEV₁/FVC (71.9%±5.4%) were lower than normal group’s FEV_1.0 (3.2±0.7 L) 、FEV_1.0/FVC (79.2%±9.9%) , and were hirher than asthma group’s FEV₁ (2.5±0.8) L and FEV_1.0/FVC (68.7%±6.1%) . In induced sputum, macrophages percentage in patients with pulmonary thesaurosis (54.3%±9.7%) was higher than asthma group and normal group (48.4±6.4, 51.9±7.3, P<0.05) , neutrophils percentage in patients with pulmonary thesaurosis group (43.9%±7.6%) was higher than asthma group and normal group (41.3%±7.1%, 39.8%±6.9%, P<0.05) , eosinophils percentage in patients with pulmonary thesaurosis group (0.3%±0.1%) was lower than asthma group (2.8±0.3, P<0.05) , lymphocytes percentage in patients with pulmonary thesaurosis group (2.0%±0.4%) was lower than asthma group (4.7±0.6, P<0.05) ; the sputum levels of IL-8 in patients with pulmonary thesaurosis group (914.7±24.3) pg/ml was higher than asthma group and normal group (747.9%±16.8 pg/ml, (405.4±13.7) pg/ml, P<0.05) . Conclusion Macrophages and IL-8 may play an important role in the pathogenesis of airway inflammation after long-term inhalation of iron and its compounds, the airway function in patients of occupational pulmonary thesaurosis was found damaged.

Objective:To investigate the application of photoplethysmogram in analyzing the fingertip pulse amplitude volume (PAV)to evaluate the endothelial function in diagnosing coronary heart disease,and to clarify its relationship with the risk factors of cardiovascular diseases.Methods:Total 409 patients with chest pain accepted coronary angiography (CAG)were selected and diveded into positive group (CAG+)(n=288)and negative group (CAG-) (n = 121)according to angiographic results.Fingertip photoplethysmogram was used to analyze the fingertip PAV by the way of applying endothelial function diagnostic after reactive ischemia,and the relationship between the PAV value and the risk factors of coronary heart diseases was analyzed,and the critical reference value of prediction index of coronary heart disease was determined.The risk factors such as age,gender,serum total cholesterol (TC ), low density lipoprotein (LDL ), high density lipoprotein (HDL ), non-HDL, serum triglyceride (TG),hypertension,diabetes,smoking,family history of coronary heart disease,body mass index (BMI)of the subjects in various groups were analyzed,and the relationship between the risk factors of coronary heart disease and PAV was analyzed by Logistic regression analysis.Results:The PAV of patients in CAG+group was significantly lower than that in CAG-group (P <0.01).The peak point of PAV was<1.37,if PAV<1.37 was used to predict the coronary heart disease,the predictive sensitivity,the specificity,the positive prediction and the negative prediction were 74.65%,44.63%, 76.24%, and 42.52%.The Logistic regression analysis showed that PAV was negatively associated with hypertension,smoking history,TG (OR= 1.476,OR=2.002, OR = 1.844;P < 0.01 ). Conclusion: PAV is associated with coronary heart disease and its risk factors (hypertension,smoking history,TG),and PAV=1.37 can be used as the peak point to predict the coronary heart disease.

BACKGROUND AND OBJECTIVETransient receptor potential canonical (TRPC) proteins, a group of Ca2' permeable nonselective cation channels, are thought to constitute store-operated calcium channels (SOCC) and mediate store-operated calcium entry (SOCE) in various cell types. Members of TRPC have been found to be involved in abnormal proliferation, differentiation, and growth of cancer cells. The aim of this study is to detect the mRNA and protein expression of TRPC in non-small cell lung cancer (NSCLC).

METHODSReal-time quantitative PCRwas performed to screen the expression of TRPC mRNA in NSCLC tissue. Protein expression of TRPC was detected by Western blot.

RESULTSAmong the seven family members of TRPC so far identified (TRPC1-7), we detected the expression ofTRPC1, TRPC3, TRPC4, TRPC6 mRNA in 24 cases of NSCLC tissue; TRPC2, TRPC5 and TRPC7 mRNA were not detectable. The relative abundance of the expressed TRPC was TRPC1 approximately equal TRPC6 > TRPC3 > TRPC4. Western blot confirmed the protein expression of TRPC1, TRPC3, TRPC4 and TRPC6 in NSCLC tissue.

CONCLUSIONOut of the seven members of TRPC, we found TRPC1, TRPC3, TRPC4, TRPC6 mRNA and protein were selectively expressed in human NSCLC tissue. This study could provide a basis for future exploration of the individual role of these TRPC proteins in mediating SOCE and in the progression of lung cancer.

Adult ; Aged ; Blotting, Western ; Calcium ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; Female ; Humans ; Lung Neoplasms ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; analysis ; TRPC Cation Channels ; genetics ; physiology

Objective To investigate the expression and significance of endothelin-1 ET-1 in urothelial transitional cell carcinoma. Methods Immunohistochemistry was applied to detect ET-1 expression in 30 cases of urothelial transitional cell carcinoma and 20 cases of normal urothelial transitional cell samples, the results were analyzed in combination with clinical data. Results ET-1 expression in urothelial transitional cell carcinoma was 40.5%, which was significantly higher than that in normal urothelial transitional cell samples(P ＜0.05). ET-1 expression was correlated with grade and stage of urothelial transitional cell carcinoma (P ＜0.05). Conclusion The over expression of ET-1 in urothelial transitional cell carcinoma tissues correlates closely with cancer genesis progression and invasiveness, suggesting that ET-1 might be a biomarker and as a therapeutic target in human urothelial transitional cell carcinoma.