Dipeptydil-peptidase-4 (DPP-4) inhibitors are designed as oral anti-hyperglycemic drugs for the treatment of type 2 diabetes owing to their effects on the incretin system through sparing incretin hormones including glucagon-like peptide-1 (GLP-1) from rapid degradation by DPP-4.It has been shown that GLP-1 signaling may exert beneficial effects on cardiovascular system.In addition to GLP-1,DPP-4 physiologically cleaves cytokines,chemokines,and neuropeptides involved in inflammation,immunity,and vascular function.It means that DPP-4 inhibitors hold promise for cardiovascular protection independent of GLP-1.And its modulation of endothelial progenitor cells,infiammatory pathway,ischemic response,and lipid metabolism emerges as the major cardiovascular target of DPP-4 inhibitors.

Proinsulin(PI)_(AUC) increased,and insulin action index,HOMA-? and △I_(30)/△G_(30) decreased progressively from NGT(n=131) to IGT(n=120) to T2DM(n=107).Obese group had higher HOMA?,Ins_(AUC),C-P_(AUC) and PI_(AUC) than did the non-obese group.

411 subjects were divided into six groups according to their FPG levels.And OGTT was carried out.Result showed that along with the FPG increasing, HOMA ?-cell index and ?I30 /?G30 were decreased progressively, and the fasting serum proinsulin level and ratio of proinsulin /insulin were increased markedly.The area under insulin curve decreased gradually when FPG was above 8 mmol/L. In conclusion, FPG is a reflection of islet ? cell function.

Aim ① To observe the relationship between TGF?_1 and diabetic nephropathy in experimental rats;② To explore the effects of ACEI-fosinopril and ATRA-losartan on renal TGF?_1 in diabetic rats and their renoprotective mechanism. Method Four groups of rats were studied, A group:normal control rats;B group: strephtozotocin induced diabetic rats;C group:diabetic rats treated with fosinopril;D group: diabetic rats treated with losartan.Blood glucose, urinary excretion rates of albumin, TGF?_1,as well as the expressions of TGF?_1 protein and TGF?_1 mRNA in renal cortex and the relative kidney were measured. Result ① The urinary excretion rates of albumin and TGF?_1 in B,C ,D groups were significant higher than those in A group, fosinopril and losartan can decrease the urinary excretion rates of the two proteins but can’t make the rate normal. ② The expressions of TGF?_1 protein in renal cortex in B group was much higher than that in other 3 groups, fosinopril and losartan can inhibit the expression of TGF?_1. ③ The expressions of TGF?_1 mRNA in renal cortex increased greatest in B group, fosinopril and losartan can low the expression. Conclusion The overexpression of TGF?_1 protein and mRNA in renal cortex of diabetic rats may be one of the mechanisms of diabetic nephropathy. Fosinopril and losartan can suppress the expressions of TGF?_1 protein and mRNA in renal cortex, decrease the urinary excretion rates of TGF?_1. So alleviate the patholochanges in kidney.

Objective To discuss the effects of the changes of sex hormone levels on quality of life and nutritional status in middle-aged and elderly hospitalized men.Methods A total of 150 male patients (aged 45-90 years) were divided into 3 groups according to age:45-59-year (the middle-aged group),60-79 year (the elderly group),and over 80 year (the old old group) (n=50,each group).Levels of serum testosterone,follicle stimulating hormone (FSH),and luteinizing hormone (LH)were detected.The questionnaire (SF-36) and micro-nutrient assessment table (MNA) were conducted.Results Serum testosterone level was significantly lower in the old old group than in the elderly group and middle-aged group [(10.5±2.5) pmol/L vs.(13.1±2.0) pmol/L and (15.6±3.2) pmol/L,respectively,P＜ 0.05].Levels of FSH and LH were significantly higher in the old old group than in the elderly group and middle-aged group [(15.5±7.7)U/L vs.(12.7±5.1)U/L and (9.5±2.9)U/L,(9.8±3.7)U/L vs.(7.8±3.2)U/L and (5.4±2.0) U/L,respectively,P＜0.05].The 8 health latitude scores of SF-36 scale and nutritional status scores were significantly lower in the old old group than in the elderly group and middle-aged group (all P＜0.05).Relation analysis sho wed that age had a negative correlation with serum testocterone level (r=-0.389,P＜0.05),and had positive correlations with the levels of serum FSH and LH (r=0.427,r=0.520,both P＜0.05).After adjusting for age,serum testocterone level was positively correlated with physical functioning,role of physical activity,body pain,general health,vitality,mental health and nutritional status in SF-36 health scale (all P＜0.05),and had no obvious correlation with social function and emotional function health dimensions (all P＞0.05).Conclusions The sex hormone levels are changed with age,which can seriously affect the health state in middle-aged and elderly hospitalized men.

Objective To observe the effects of metformin on expression of Adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK),nuclear factor-κB (NF-κB) and transforming growth factor β1 (TGF-β1) in cultured rat glomerular mesangial cells (MCs),and explore its renoprotective mechanisms.Methods MCs were cultured in the medium with normal glucose (group NG,5.6 mmol/L),high glucose (group HG,25mmol/L) and different concentrations of metformin (group M1,M2,M3).After 48 h exposure,the supernatants and MCs were collected.The expression of NF-κB and TGF-β1 mRNA was analyzed by real time-PCR.Total-AMPK,phospho-Thr-172 AMPK (p-AMPK),NF -κB p65 and TGF-β1 were visualized by Western blot.Results The real time-PCR and Western blot result showed MCs could express AMPK,NF-κB and TGF-β1 mRNA and protein.After stimulated by HG,the levels of intracellular NF-κB and TGF-β1 expressions were significantly increased compared with group NG (P ＜ 0.05); The levels of NF-κB and TGF-β1 were significantly decreased in group M1,M2 and group M3 compared with group HG in a dose-dependent manner.After stimulated by HG,the level of intracellular p-AMPK were down-regulated compared with group NG(all P ＜ 0.05); The expression of p-AMPK increased with the rising of metformin concentration,presenting the opposite trend (P ＜ 0.05),while the level of total-AMPK protein was unchanged with exposure to HG or different concentrations of mefformin(P ＞ 0.05).Conclusion Metformin can suppress the expression of NF-κB and TGF-β1 of glomerular MCs induced by HG via AMPK activation,which may partly contribute to its reno-protection.

Aims To observe the influences of met-formin ( MET ) on the expression of renal tissue ad-vanced glyclation end-products( AGEs) protein and its receptor mRNA ( RAGE mRNA ) in type 2 diabetes mellitus (T2DM) model rats, and to discuss the mech-anism of the MET in the treatment of diabetic nephrop-athy ( DN ) . Methods The rat model of T2 DM was established by fed with high-fat diet and intraperitoneal injection of low-dose of streptozotocin ( STZ ) . All rats were randomly divided into metformin group( MET,300 mg·kg-1 ·d-1 ) , glyburide group( GLY,5 mg·kg-1 ·d-1),T2DM model group(T2DM) and normal con-trol group ( NC ) . After 8 weeks ’ observation, blood glucose ( BG ) , glycated hemoglobin ( HbA1 c ) , blood urea nitrogen(BUN), urinary albumin,urinary AGEs and urine creatinine were detected. The expression of renal tissue AGEs was detected by immunohistochemis-try assay, and the expression of RAGE mRNA was measured by real-time PCR. Results The levels of BG, HbA1c , urinary albumin/urine creatinine ( UACR ) , glomerular basement membrane thickness ( GBMT ) in MET group and GLY group were signifi-cantly lower than those of T2DM group, while higher than those of NC group(P<0. 05), the levels of BG, FINS and HbA1 c were not statistically significant be-tween MET and GLY group ( P >0. 05 ) . The urinary AGEs/urine creatinine( UGCR) , the expressions of re-nal tissue AGEs and RAGE mRNA in MET group and GLY group were significantly decreased compared with those of T2 DM group ( P < 0. 05 ) , but higher than those of NC group ( P <0. 05 ) . The UGCR, the ex-pressions of AGEs and RAGE mRNA in MET group were lower than those of GLY group(P<0. 05). Con-clusion MET can reduce the accumulation of AGEs in the renal tissue,and down-regulate the over-expres-sion of RAGE mRNA in T2DM rats.

Objective: To explore the mechanism of Wuling Powder in the treatment of chronic heart failure (CHF) based on network pharmacology. Methods: Traditional Chinese Medicine Systems Pharmacology Database (TCMSP) and literature mining were used to search the chemical components and targets of Wuling Powder, and a single drug-active ingredients-target network was established. The related targets of chronic heart failure were collected through Genecards and OMIM databases, the network model of active components-CHF-targets was constructed and analyzed by Cytoscape 3.7.1 software. The protein-protein interaction (PPI) network was constructed by STRING database platform, and the gene oesthetics (GO) function annotation and Kyoto Encyclopedin of Genes and Genomes (KEGG) signal pathway enrichment analysis were performed by DAVID online tools. The molecular docking was performed using Surflex software. Results: Fifty components, 29 potential targets, 8 243 targets related to chronic heart failure, and 27 targets of Wuling Powder-CHF were obtained. The network analysis results showed that the key targets of Wuling Powder in the treatment of chronic heart failure included CASP3, RELA, AR, ESR1, CHRM1 and CASP8, etc. Biological processes mainly involved signal transduction, nervous system development, transcription initiation from RNA polymerase II promoter in response to estradiol, synaptic transmission, cholinergic synaptic transmission, etc. KEGG enrichment involved neuroactive ligand-receptor interaction, PI3K-Akt signaling pathway, cholinergic synapse, etc. The molecular docking results showed that (+)-catechin, taxifolin and other key compounds in Wuling Powder had better binding ability with key targets such as CASP8, CHRM1, and NR3C1. Conclusion: The material basis and mechanism of Wuling Powder in the treatment of chronic heart failure were revealed based on network pharmacology, which provided a certain theoretical basis for the clinical application of Wuling Powder.

BACKGROUNDHepatitis B virus (HBV) x protein (HBx) in HepG2 cells causes a moderate decrease in proteolysis activity of the proteasome. A highly conserved Kunitz-type serine protease inhibitor domain within 154 amino acid residues of HBx has been identified. In this study, a peptide chain derived from the Kunitz domain (PKD) was used to study its effect on the cell cycle and apoptosis of HepG2 cells, and investigated the function of PKD on the activities of proteasomes and AAA-ATPase p97, which involves in the ubiquitin-proteasome protein degradation pathway.

METHODSThe PKD peptide (Phe-Val-Leu-Gly-Gly-Cys-Arg-His-Lys) was chemically synthesized. MTT assays were used to determine the effects of PKD on HepG2 cell growth. Mouse anti-p97 antibody was developed for Western blotting to detect the expression of p97. ATPase activity of proteasomes was measured using a colorimetric assay. Peptidase activities of proteasomes were analyzed with various peptidase-specific fluorogenic peptide substrates. Flow cytometry was used to determinate cell cycle phase and apoptosis.

RESULTSViability of HepG2 cells decreased in a PKD-dose-dependent manner. Cells exhibited significant cytotoxicity in the presence of 15 mmol/L of PKD. Western blotting analysis showed that expression of p97 was suppressed in HepG2 cells treated with PKD compared to untreated cells. The ATPase activity of proteasomes from immunoprecipitates of HepG2 cells pretreated with PKD was apparently decreased. Chymotryptic activity of proteasomes in HepG2 cells was significantly inhibited by 10 mmol/L PKD; tryptic activity and peptidylglutamyl peptide hydrolase activity of proteasomes were less inhibited by PKD than chymotryptic activity. The cell cycle phase of HepG2 cells treated with PKD for 36 hours was blocked largely at the G(0)-G(1) phase, while untreated control cells were mainly in S phase. PKD also significantly induced apoptosis.

CONCLUSIONSThe peptide derived from Kunitz domain of HBx protein induces HepG2 cell growth arrest and apoptosis, which may result from down-regulation of p97 expression, and decrease of both the ATPase and chymotryptic activities of proteasomes.

Adenosine Triphosphatases ; metabolism ; Animals ; Apoptosis ; drug effects ; Blotting, Western ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Enzyme Activation ; drug effects ; Humans ; Lipopeptides ; chemistry ; pharmacology ; Mice ; Nuclear Proteins ; metabolism ; Trans-Activators ; chemistry ; Viral Regulatory and Accessory Proteins ; chemistry

Objective To understand the genetic basis of the adaptation of Culex tritaeniorhynchus to different environmental ecology in Jining City, Shandong Province, so as to provide insights into understanding of the population structure or isolation pattern of Cx. tritaeniorhynchus in the city. Methods Seven sampling sites were selected from urban, suburban and rural areas of Jining City, Shandong Province from June to August 2023, and mosquitoes were collected using mosquito-trapping lamps. All collected adult mosquitoes were identified morphologically. Genomic DNA was extracted from a single female Cx. tritaeniorhynchus mosquito, and the mitochondrial cytochrome C oxidase I (COI) gene was amplified using a PCR assay, sequenced and subjected to molecular identification. The number of haplotypes, haplotype diversity (Hd), nucleotide diversity (Pi), and average number of nucleotide differences (K) of Cx. tritaeniorhynchus DNA sequences were estimated among different sampling sites using the software DnaSP 6, and a neutrality test was performed. The fixation index (F_ST), and gene flow (number of migrants, Nm) of Cx. tritaeniorhynchus populations were calculated using the software Arlequin 3.5.2, and subjected to analysis of molecular variance (AMOVA). In addition, a haplotype network diagrams and a phylogenetic tree of Cx. tritaeniorhynchus populations were created using the software PopART and MEGA 11, respectively. Results A total of 420 sequences were successfully amplified from the COI gene of Cx. tritaeniorhynchus samples collected from 7 sampling sites in Jining City, and a gene fragment sequence with a length of 603 bp was obtained, with 55 variable sites and 46 haplotypes and without insertion or deletion mutations. Of the 46 haplotypes, H01 was the dominant shared haplotype, and the haplotype frequency increased gradually from urban areas (34.00%) to rural areas (47.00%). The mean Hd, Pi and K values of Cx. tritaeniorhynchus COI genes were 0.814, 0.024 and 14.129, 0.489, 0.016 and 7.941 and 0.641, 0.016 and 10.393 in suburban, urban, and rural areas, respectively, with the highest population diversity of Cx. tritaeniorhynchus in suburban areas and the lowest in urban areas. Paired F_ST analysis among different types of sampling sites showed that the mean F_ST value was 0.029 between urban and suburban areas, indicating more frequent inter-population communication. AMOVA revealed that the percentage of intra-population variation (95.74%) was higher than that of inter-population variation (4.26%). Neutrality tests showed deviation from neutrality in Cx. tritaeniorhynchus populations collected from Nanyang Township (Tajima’s D = 2.793, Fu’s Fs = 6.429, both P values < 0.05). In addition, the mismatch distribution curves of Cx. tritaeniorhynchus COI gene appeared bimodal or multimodal patterns in Jining City, indicating a relatively stable overall population size. Conclusions The mitochondrial COI gene may be used as a molecular marker to investigate the population genetic diversity of Cx. tritaeniorhynchus. The population genetic diversity of Cx. tritaeniorhynchus is higher in the suburban areas of Jining City than in rural and urban areas, and there are frequent genetic exchanges between Cx. tritaeniorhynchus populations from urban and suburban areas.