AIM:To explore the influence of microRNA-193b-5p(miR-193b-5p)on apoptosis of cardiomyo-cytes in an ischemia-hypoxia(IH)environment and the possible mechanism.METHODS:Human AC16 cardiomyocytes were cultured in vitro,and an IH model of cardiomyocytes was established.The cardiomyocytes were divided into control group,IH group,IH+miR-193b-5p inhibitor group,and IH+inhibitor NC group.The cells in control group were regularly cultured,those in IH group were treated with a hypoxia chamber for 24 h to induce cardiomyocyte apoptosis,while those IH+miR-193b-5p inhibitor and IH+inhibitor NC groups were transfected with corresponding plasmids by the same opera-tion method and then underwent IH treatment for 24 h to induce cardiomyocyte apoptosis.RT-qPCR was used to detect the expression of miR-193b-5p in cardiomyocytes after IH.The viability of cardiomyocytes was detected by CCK-8 method.Lactate dehydrogenase(LDH)was detected to understand the damage of cardiomyocytes.The apoptotic rate was detected by flow cytometry.The protein levels of Bax,Bcl-2 and cleaved caspase-3 were detected by Western blot.Finally,down-stream target genes were predicted by RNA sequencing combined with bioinformatics methods,and the interaction relation-ship between miR-193b-5p and RING(really interesting new gene)finger protein 4(RNF4)gene was verified by RT-qPCR and Western blot experiments.RESULTS:Compared with control group,miR-193b-5p was highly expressed in cardio-myocytes with IH.Furthermore,in the IH environment,the viability of cardiomyocytes decreased,and cell damage and cell apoptosis increased,while inhibiting the expression level of miR-193b-5p could enhance the viability of cardiomyo-cytes,reduce cell damage,and alleviate the apoptosis of cardiomyocytes induced by IH.The results of RNA sequencing and further experiments verified that miR-193b-5p might act on the RNF4 gene to exert its effect.CONCLUSION:Inhi-bition of miR-193b-5p can alleviate the IH injury and apoptosis of cardiomyocytes.The mechanism might be that miR-193b-5p inhibition exerts a cardioprotective effect against apoptosis by mediating the expression of the RNF4 gene.

Objective To investigate the impact of mitophagy,mediated by the long non-coding RNA SNHG16(LncRNA SNHG16)on diabetes-associated cognitive impairment(DCI).Methods 29 male C57BL/J mice were randomly divided into normal control(NC)group,DCI group and DCI+mitochondrial autophagy inhibitor(DCI+Mdivi-1)group.Morris water maze and new object recognition test were used to detect the cognitive function of mice,qRT-CPR was used to detect the expression of LncRNA SNHG16 and mitochondrial autophagy marker mRNA.Western blot were used to detect the expression of related protein.The mouse hippocampal neurons HT22 were divided into control(Con)group,high glucose(HG)group,HG+SNHG16 silencing(HG+sh-SNHG16)group and HG+no-load control(HG+sh-NC)group.CCK8 method and lactate dehydrogenase(LDH)method were used to detect neuronal damage.JC-1 method was used to detect mitochondrial membrane potential.Results Compared with NC group,the expression of LncRNA SNHG16 and the expression of autophagy-related gene 5,PTEN-induced putative kinase 1(PINK1),Parkin and microtubule associated protein light chain 3(LC3)Ⅱ/Ⅰ increased(P<0.05),while the expression of mitochondrial autophagy-related proteins P62 and mitochondrial outer membrane transposase 20(TOMM20)decreased in T2DM group.Compared with DCI group,the cognitive dysfunction of mice improved,and the expression level of LncRNA SNHG16 decreased in the DCI+Mdivi-1 group(P<0.05).The expressions of LncRNA SNHG16,LC3 Ⅱ/Ⅰ,PINK1 and Parkin were higher in HG group than in Con group(P<0.05),while the cell survival rate and TOMM20 protein expression were lower in HG group than in Con group(P<0.05).Silence of LncRNA SNHG16 can restore the activity of HT22 cells and mitochondrial membrane potential,and reduce the level of mitochondrial autophagy under HG condition.Conclusions The expression level of LncRNA SNHG16 was up-regulated in the hippocampus brain region of mice with diabetic cognitive dysfunction,and mitophagy was overactivated.Silencing of LncRNA SNHG16 inhibits mitophagy in hippocampal neurons and alleviates HG induced hippocampal neuronal damage.

Objective To investigate the impact of mitophagy,mediated by the long non-coding RNA SNHG16(LncRNA SNHG16)on diabetes-associated cognitive impairment(DCI).Methods 29 male C57BL/J mice were randomly divided into normal control(NC)group,DCI group and DCI+mitochondrial autophagy inhibitor(DCI+Mdivi-1)group.Morris water maze and new object recognition test were used to detect the cognitive function of mice,qRT-CPR was used to detect the expression of LncRNA SNHG16 and mitochondrial autophagy marker mRNA.Western blot were used to detect the expression of related protein.The mouse hippocampal neurons HT22 were divided into control(Con)group,high glucose(HG)group,HG+SNHG16 silencing(HG+sh-SNHG16)group and HG+no-load control(HG+sh-NC)group.CCK8 method and lactate dehydrogenase(LDH)method were used to detect neuronal damage.JC-1 method was used to detect mitochondrial membrane potential.Results Compared with NC group,the expression of LncRNA SNHG16 and the expression of autophagy-related gene 5,PTEN-induced putative kinase 1(PINK1),Parkin and microtubule associated protein light chain 3(LC3)Ⅱ/Ⅰ increased(P<0.05),while the expression of mitochondrial autophagy-related proteins P62 and mitochondrial outer membrane transposase 20(TOMM20)decreased in T2DM group.Compared with DCI group,the cognitive dysfunction of mice improved,and the expression level of LncRNA SNHG16 decreased in the DCI+Mdivi-1 group(P<0.05).The expressions of LncRNA SNHG16,LC3 Ⅱ/Ⅰ,PINK1 and Parkin were higher in HG group than in Con group(P<0.05),while the cell survival rate and TOMM20 protein expression were lower in HG group than in Con group(P<0.05).Silence of LncRNA SNHG16 can restore the activity of HT22 cells and mitochondrial membrane potential,and reduce the level of mitochondrial autophagy under HG condition.Conclusions The expression level of LncRNA SNHG16 was up-regulated in the hippocampus brain region of mice with diabetic cognitive dysfunction,and mitophagy was overactivated.Silencing of LncRNA SNHG16 inhibits mitophagy in hippocampal neurons and alleviates HG induced hippocampal neuronal damage.

AIM:To explore the influence of microRNA-193b-5p(miR-193b-5p)on apoptosis of cardiomyo-cytes in an ischemia-hypoxia(IH)environment and the possible mechanism.METHODS:Human AC16 cardiomyocytes were cultured in vitro,and an IH model of cardiomyocytes was established.The cardiomyocytes were divided into control group,IH group,IH+miR-193b-5p inhibitor group,and IH+inhibitor NC group.The cells in control group were regularly cultured,those in IH group were treated with a hypoxia chamber for 24 h to induce cardiomyocyte apoptosis,while those IH+miR-193b-5p inhibitor and IH+inhibitor NC groups were transfected with corresponding plasmids by the same opera-tion method and then underwent IH treatment for 24 h to induce cardiomyocyte apoptosis.RT-qPCR was used to detect the expression of miR-193b-5p in cardiomyocytes after IH.The viability of cardiomyocytes was detected by CCK-8 method.Lactate dehydrogenase(LDH)was detected to understand the damage of cardiomyocytes.The apoptotic rate was detected by flow cytometry.The protein levels of Bax,Bcl-2 and cleaved caspase-3 were detected by Western blot.Finally,down-stream target genes were predicted by RNA sequencing combined with bioinformatics methods,and the interaction relation-ship between miR-193b-5p and RING(really interesting new gene)finger protein 4(RNF4)gene was verified by RT-qPCR and Western blot experiments.RESULTS:Compared with control group,miR-193b-5p was highly expressed in cardio-myocytes with IH.Furthermore,in the IH environment,the viability of cardiomyocytes decreased,and cell damage and cell apoptosis increased,while inhibiting the expression level of miR-193b-5p could enhance the viability of cardiomyo-cytes,reduce cell damage,and alleviate the apoptosis of cardiomyocytes induced by IH.The results of RNA sequencing and further experiments verified that miR-193b-5p might act on the RNF4 gene to exert its effect.CONCLUSION:Inhi-bition of miR-193b-5p can alleviate the IH injury and apoptosis of cardiomyocytes.The mechanism might be that miR-193b-5p inhibition exerts a cardioprotective effect against apoptosis by mediating the expression of the RNF4 gene.

AIM:This study aimed to investigate the effects of hsa-miR-204-5p on the viability,migration,cell cycle,and apoptosis of human vascular endothelial cells.METHODS:We established a model using the hsa-miR-204-5p mimic in the human umbilical vein endothelial cell line EA.hy926.We evaluated the effects of hsa-miR-204-5p on endothelial cell functionality through various analyses,including cell scratch,Transwell,CCK-8,cell cycle,and apopto-sis assays.Subsequently,we employed RNA sequencing and RT-qPCR to predict and verify the downstream target genes of hsa-miR-204-5p.Genes meeting the criteria of log2FC≤-0.5 and P＜0.05 in RNA sequencing and those predicted as downstream target genes of hsa-miR-204-5p by the miRWalk database were intersected.Furthermore,we conducted Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.RESULTS:Overexpres-sion of hsa-miR-204-5p inhibited the viability and migration of EA.hy926 cells,and reduced their apoptotic rate and the proportion of cells in S phase.Enrichment analyses showed that downstream target genes of hsa-miR-204-5p,including MAPT,PPP3R1,PRKACB,PTPRR,MAP2K4,CACNA2D2 and RPS6KA6,exhibited enrichment in MAPK signaling pathway.RT-qPCR results revealed that the mRNA expression levels of MAPT and MAP2K4,especially MAPT,were sig-nificantly down-regulated after overexpression of hsa-miR-204-5p.CONCLUSION:The findings suggest that hsa-miR-204-5p suppresses the biological behaviors of endothelial cells,such as viability,migration,and apoptosis,likely through the inhibition of MAPT/MAPK signaling pathway.

Objective To study the effect of mitogen-activated protein kinas/extracellular signalregulated kinase (MAPK/ERK) signaling pathway on cell proliferation modulated by Sonic Hedgehog (Shh) signaling in fibroblast-like synoviocytes (FLS) isolated from patients with active rheumatoid arthritis (RA).Methods The synovial tissue were collected by the synovial arthroscopic debridement or arthroscopic synovectomy of RA patients with active disease activity [disease activity score(DAS)28 ≥3.2].The RA-FLS were primarily cultured by the explanted culture,and then were treated with Shh agonist purmorphamine,inhibitor cyclopamine or MAPK/ERK signaling pathway inhibitor U0126,respectively.Western blotting was used to examine the phosphorylation level of ERK 1/2 (p-ERK1/2),which was the critical protein of MAPK/ERK signaling.The cell proliferation activity was detected using cell proliferation and cytotoxicity kit-8 (CCK8),and the cell proliferation rate was detected using a flow cytometry.Analysis of variance and Kruskal-Wallis H(K) test were used for statistical analysis.Results Compared with the control group,purmorphamine transiently increased p-ERK1/2 protein at the concentration of 1 μmol/L,and the peak activations of p-ERK1/2 took place at 15 min (P＜0.01).Cyclopamine and U0126 decreased the expression ofp-ERK1/2 protein (P＜0.01).After the RA-FLS treated with purmorphmine(1 μmol/L)for 48 hours,the cell proliferation activity was (114±4)％ and the percentage of S phase cells was (8.39±0.60)％,which was significantly higher than those of the control group (100±0)％ (P＜0.01) and (3.29±0.69)％ (P＜0.01).After treated with cyclopamine (10 μmol/L) for 48 hours,the cell proliferation activity of RA-FLS was (89±1)％ (P＜0.05) and the percentage of S phase cells was (1.53±0.22)％ (P＜0.05).When co-treated with purmorphamine (1 μmol/L) and U0126 (10 μmol/L),the cell proliferative activity was (89±2)％ (P＜0.05) and the percentage of S phase cells was(1.07±0.25)％(P＜ 0.05).Conclusion Shh may promote proliferation of RA-FLS via modulating MAPK/ERK signaling,which in turn contributes to hyperplasia of synovium and ultimately leading to RA.

Objective To explore the telomere length (TL) in buccal cells and its possible implications for long-lived families of Zhuang nationality in Bama area of Guangxi.Methods Relative TL in buccal cells from Bama long-lived families (BLF,n=1250) was determined by real time PCR and compared between Bama non-long-lived families (BNLF,n=556) and Pingguo longlived families (PLF,n=630).All participants were Zhuang ethnic.Results The TL in buccal cells was negatively correlated with age (R=-0.215,P=0.000) and was independent of sex in all subjects (n=2436).There were no significant differences in TL between males and females in different of ages in BLF (all P＞0.05).Similar tendency was observed in most but not all age groups in the two other families.The TL was significantly longer in BLF aged over 65 years (the first offspring of the longlived individual) than in BNLF aged over 65 years (1.969 vs.1.622,P=0.004) and the TL of BLF aged over 90 years was comparable to that of BNLF aged over 65 years (1.662 vs.1.622,P=0.955),which indicating that the offspring of long-lived individuals inherited longer TL from their parental generation.Conclusions Telomere length is shorten with aging in long-lived families in Bama area.The TL of BLF has a tendency of inheritance,which may be one of the mechanisms of longevity in Bama area.

The levels of total cholesterol, high-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol, apolipoprotein (APO) A1 and APO B were lower in Bai Ku Yao than those in Han nationalities (all P<0.01). There was no significant difference in serum triglyceride levels and the ratio of Apo A1 to Apo B between two nationalities. Dyslipidemia was positively correlated with body mass index, waist circumference, total energy and total fat intakes, and inversely correlated with degree of physical activity and total dietary fiber intake in both ethnic groups. In addition, dyslipidemia was also positively correlated with age and alcohol consumption in Han, but not in Bai Ku Yao.

Objective To investigate the prevalence rate of hyperlipidemia in the middle-aged and elderly in Guangxi Hei Yi Zhuang population. Methods A total of 657 people of Hei Yi Zhuang nationality aged 40 years and over were surveyed. Blood pressure, body height, body weight, serum lipid and apolipoprotein levels were measured, and both body surface areas and body mass index were calculated. The results were compared with those in 520 people of Han nationality who also live in that district. Results The prevalence of hypercholesterolemia, hypertriglyceridemia, and hyperlipidemia in Hei Yi Zhuang vs. Han population were 28.9% vs. 35.8%(P0.05), and 36.2% vs. 42.3%(P0.05) respectively. Conclusions Prevalence of hyperlipidemia of Hei Yi Zhuang is lower than that of Han ethnic group, which might results from different dietary habit, life style, physical activity, and even genetic background.

Objective To investigate the role of thyroid autoimmunity in chronic urticaria. Methods The thyroid function, anti-thyroid auto-antibodies, and relevant cytokines were detected by RIA and ELISA methods in 56 patients with chronic urticaria and 40 healthy controls. Results Serum thyroid-stimulating hormone (TSH), anti-thyroglobulin antibody (TG-Ab) and anti-thyromicrosome antibody (TMA ) levels were increased in 8, 5 and 7 cases, respectively, in 56 patients, which were significantly higher than those in healthy controls (P