Objective:To analyze the clinical characteristics, prognosis and gene mutation in patients with dilated cardiomyopathy (DCM) in Keshan disease area of Sichuan Province, and to explore the risk factors for all-cause death in DCM patients.Methods:In June 2016, 55 DCM patients diagnosed at the local disease prevention and control center through clinical manifestations, electrocardiogram examination, and echocardiography were selected as the survey subjects in Mianning County, Liangshan Yi Autonomous Prefecture, and Renhe District, Panzhihua City, Keshan disease areas of Sichuan Province. Baseline clinical data were analyzed and long-term follow-up was conducted. The follow-up period ended June 15, 2021, with the endpoint of all-cause death. Univariate Cox regression was used to analyze the influencing factors of all-cause death in patients, and Kaplan-Meier (K-M) survival curve was used to analyze the survival time of patients. At the same time, peripheral venous blood was collected from 27 DCM patients. After separating white blood cells, DNA was extracted, and whole exome sequencing was performed to screen potential pathogenic genes.Results:Among the 55 DCM patients, 40 were males and 15 were females. The age was (54.09 ± 12.38) years old. The heart function classification of New York Heart Association (NYHA) was mainly grade Ⅱ and Ⅲ, accounting for 94.55% (52/55). The follow-up time for 55 DCM patients was (7.02 ± 2.96) years, and 17 patients experienced all-cause death, accounting for 30.91% (17/55), including 15 males and 2 females. Compared with the survival group, the death group had a lower incidence of syncope (χ 2 = 6.57, P = 0.010), but higher rates of bilateral lower limb edema (χ 2 = 6.43, P = 0.017), pulmonary congestion (χ 2 = 7.61, P = 0.006), intraventricular conduction block (χ 2 = 6.41, P = 0.011), and angiotensin-converting enzyme inhibitor (ACEI) use (χ 2 = 6.57, P = 0.010), as well as increased left ventricular diameter ( t = 2.36, P = 0.022). Univariate Cox regression analysis showed that bilateral lower limb edema [hazard ratio ( HR) = 4.61, P = 0.042] and intraventricular conduction block ( HR = 3.20, P = 0.019) were risk factors for all-cause death of DCM patients. The results of K-M survival curve analysis showed that patients with bilateral lower limb edema and intraventricular conduction block had higher all-cause death rates (log-rank χ 2 = 5.02, 6.24, P = 0.025, 0.012). Whole exome sequencing results showed that 4 patients were detected to carry pathogenic or suspected pathogenic gene mutations, with a positive rate of 14.81% (4/27), involving three genes: β-myosin heavy chain 7 (MYH7), calreticulin 3 (CALR3), and gelsolin (GSN). Conclusions:The all-cause death rate of DCM patients in the Keshan disease area of Sichuan Province is relatively high. Dead patients are prone to bilateral lower limb edema, pulmonary congestion, and intraventricular conduction block, as well as increased left ventricular diameter. Bilateral lower limb edema and intraventricular conduction block are independent predictive risk factors for all-cause death in DCM patients. MYH7, CALR3 and GSN are involved in the pathogenesis of DCM.

Objective: To investigate the correlation between type 2 diabetes mellitus (T2DM) and hepatocellular carcinoma (HCC) by case-control study. Methods: From January 2006 to December 2015, the data of 1 350 first diagnosis HCC patients at the First Affiliated Hospital of Zhengzhou University were collected and analyzed. The univariate and multivariate logistic regression were performed to analyze the risk factors of HCC genesis, and the risk factors were further stratified. Results: The results of univariate logistic regression analysis showed that smoking and obesity were not risk factors of HCC genesis (both P>0.05). Alcoholism, hepatitis B virus (HBV) infection, hepatitis C virus (HCV) infection, T2DM, and family history of HCC were the risk factors of HCC genesis (odds ratio (OR)=2.593, 4.583, 3.732, 1.955 and 1.622, 95% confidence interval (CI) 1.883 to 3.549, 3.026 to 6.940, 2.544 to 5.367, 1.708 to 2.477 and 1.314 to 2.267, all P<0.05). The results of multivariate logistic regression analysis indicated that alcoholism, HBV infection, HCV infection, T2DM and family history of HCC were independent risk factors of HCC genesis (OR=2.034, 4.564, 2.831, 1.881 and 1.616, 95%CI 1.507 to 2.745, 2.672 to 7.765, 1.562 to 5.047, 1.314 to 2.671 and 1.177 to 2.228, all P<0.01), and T2DM could increase the risk of HCC genesis (P<0.01). In order to exclude the interference of alcoholism, HBV infection and HCV infection on above results, further stratified analysis showed that T2DM was one of the independent risk factors of HCC genesis (χ²=5.190, P=0.023), and had synergistic effect with alcoholism, HBV infection and HCV infection (χ²=32.848, P<0.01). There were significant differences in the propovtion, duration of disease and hemoglobin A_1c level of T2DM patients between the case group and the control group (χ²=46.618, 81.644 and 43.092, all P<0.01). Conclusions T2DM is one of the independent risk factors or HCC genesis. T2DM is correlated with the pathogenesis of HCC. The clinical desease course and blood glucose control in patients with T2DM are also correlated with the pathogenesis of HCC.

Objective To study the function and mechanism of receptor-interacting protein kinase 3 (RIPK3)-mediated necroptosis in liver ischemia-reperfusion injury (IRI) in mice.Methods Sixty mice were randomly divided into four groups using Stata statistical software:Wild-type (WT)-sham group,WT-IRI group,HKO (HKO:RIPK3 liver-specific knockout)-sham group and HKO-IRI group.Sham operation was used as a control in which only the hepatic portal blood vessels were freed after laparotomy,and blood flow was not blocked.In the WT-IRI group and the HKO-IRI group,the hepatic portal vein was freed,and the blood supply of left hepatic lobe and the mid-hepatic lobe wer blocked for 90 min,then the blood vessels were opened for 6 h.Blood and liver tissue samples of each group of mice were taken to detect liver function.Inflammatory infiltration and liver injury were detected by immunohistochemistry and hematoxylin and eosin (HE) staining,and autophagyassociated protein LC3-Ⅱ and P62 were detected by Western blotting.The primary hepatocytes of WT mice and HKO mice were extracted and divided into control group and hypoxia-reoxygenation group (HIR group).After attachment of primary hepatocytes,the HIR group was given hypoxia for 6 h and reoxygenated for 4 h.The supernatant was taken for detecting ALT and AST,and the cell extract protein was used to detect LC3-Ⅱ and P62.Results As compared with the control groups,the liver functions of the IRI groups were significantly impaired,and as compared with the WT-IRI group,the liver damage was significantly aggravated in the HKI-IRI group (P ＜ 0.05),and the LC3-Ⅱ protein content was significantly decreased and the P62 protein content was increased.Similarly,after hepatocytes were were given hypoxia and reoxygenated,HKO-derived hepatocytes were more severely damaged than WT-derived hepatocytes.Conclusions Blocking RIPK3-mediated necroptosis of hepatocytes could induce autophagy inhibition,which aggravates hepatic ischemia-reperfusion injury.

Reactive oxygen species(ROS)and immune response play critical roles in the progression of liver dis-eases.DJ-1,also known as Parkinson disease 7(Park7),is extensively expressed in cells and tissues,where it governs numerous biological functions including chaperone activity,protease function,tran-scriptional and mitochondrial regulation,and ROS modulation.Moreover,we have established that DJ-1 plays a critical role in initiating an inflammatory response by modulating ROS generation.Therefore,DJ-1 may play an important role in the progression of liver diseases by modulating ROS and the immune response.Recently,we have shown that DJ-1 deficiency negatively regulates proliferation of hepatic progenitor cells(HPCs)by impairing the formation of HPC-associated fibrosis and inflammatory niches.Deficiency of DJ-1 ameliorates liver fibrosis by inhibiting hepatic ROS production and inflammation;moreover,in a classic diethylnitrosamine(DEN)-mediated hepatocellular carcinoma(HCC)mouse model,deletion of DJ-1 ameliorates tumorigenesis and HCC cell proliferation by regulating hepatic inflammation and reducing the activity of the interleukin 6/signal transducer and activator of transcription 3(IL-6/STAT3)signaling pathway.Taken together,these data suggest a critical function for,and therapeutic value of,DJ-1 in treatment of liver diseases.The aim of this review is to summarize these functions and the underlying molecular mechanisms of DJ-1 in liver diseases,and to highlight the potential therapeutic value and future research direction of DJ-1 in liver diseases.

Objective To compare the eradication rate of helicobacter pylori between furazolidone combined with omeprazole and levofloxacin and tinidazole combined with omeprazole and levofloxacin in elderly patients with Hp positive chronic atrophic gastritis.Methods A total of 96 patients with Hp positive chronic atrophic gastritis were randomly divided into two groups,48 cases in each group.Control group was treated with tinidazole plus omeprazole and levofloxacin,while study group was treated with furazolidone plus with omeprazole and levofloxacin.The gastric function,inflammatory factors and immune function indexes were measured,and the clinical efficacy and the eradication rate of Hp were compared.Results After treatment,symptoms and pathological scores significantly decreased in both groups,G-17,PG-I contents increased significantly,PG-II content decreased significantly,serum IL-6,TNF-α,sIL-2R contents decreased significantly,CD4+ and CD4+/CD8+ contents increased significantly,and CD8+ content decreased significantly(P<0.05).There were significant differences in indexes mentioned above between the two groups(P<0.05).The effective rate and negative rate of Hp in the control group were significantly lower than those in the study group(P<0.05).Conclusion Furazolidone combined with omeprazole and levofloxacin

Objective To compare the eradication rate of helicobacter pylori between furazolidone combined with omeprazole and levofloxacin and tinidazole combined with omeprazole and levofloxacin in elderly patients with Hp positive chronic atrophic gastritis.Methods A total of 96 patients with Hp positive chronic atrophic gastritis were randomly divided into two groups,48 cases in each group.Control group was treated with tinidazole plus omeprazole and levofloxacin,while study group was treated with furazolidone plus with omeprazole and levofloxacin.The gastric function,inflammatory factors and immune function indexes were measured,and the clinical efficacy and the eradication rate of Hp were compared.Results After treatment,symptoms and pathological scores significantly decreased in both groups,G-17,PG-I contents increased significantly,PG-II content decreased significantly,serum IL-6,TNF-α,sIL-2R contents decreased significantly,CD4+ and CD4+/CD8+ contents increased significantly,and CD8+ content decreased significantly(P<0.05).There were significant differences in indexes mentioned above between the two groups(P<0.05).The effective rate and negative rate of Hp in the control group were significantly lower than those in the study group(P<0.05).Conclusion Furazolidone combined with omeprazole and levofloxacin

Objective To evaluate inhibitory effect of Chlamydia trachomatis plasmid-encoded protein pORF5 on HeLa cell apoptosis induced by tumor necrosis factor-alpha(TNF-α). Methods The recombinant lentiviral expression vector containing pORF5 gene and helper plasmids were co-transfected into 293T cells to prepare the recombinant lentivirus. Then, the lentivirus particles were collected and concentrated, and used to infect HeLa cells. Flow cytometric screening identified stable pORF5-expressing HeLa (pORF5-HeLa) cells. Meanwhile, the empty plasmid was transfected into HeLa cells to prepare control HeLa cells. The two cell lines were both divided into two subgroups to be treated with 20μg/L TNF-αand fresh culture medium respectively for 6 hours. Then, Hoechst 33258 staining was performed to observe morphological changes of apoptotic cells, flow cytometry to detect cell apoptosis, real-time PCR to measure the mRNA expression of Caspase3, Bax and Bcl-2, and Western blot analysis to determine the protein expression of Bax and Bcl-2. Results After 6-hour treatment with TNF-α, Hoechst 33258 staining showed variable degrees of karyopyknosis and karyorrhexis, and highly-refractive blue apoptotic bodies in the pORF5-HeLa cells and control HeLa cells. The pORF5-HeLa cells and control HeLa cells both showed significantly higher apoptosis rate in the treated subgroup than in the untreated subgroup (pORF5-HeLa cells:35.5%± 4.5%vs. 9.5%± 1.5%, t=13.53, P<0.01;control HeLa cells:63.6%± 5.8%vs. 7.9%± 0.9%, t=32.36, P<0.01). Compared with treated control HeLa cells, treated pORF5-HeLa cells showed significant decreases in mRNA expression of Bax(72.8%)and Caspase 3(84.5%)(t = 35.29, 42.25, respectively, both P < 0.01), as well as in Bax protein expression(t = 17.58,P < 0.01), but significant increases in Bcl-2 mRNA and protein(6.8 times)expression(t = 87.12, 18.93, respectively, both P <0.01). Conclusion pORF5 plasmid protein can inhibit TNF-α-induced HeLa cell apoptosis likely by increasing the expression of anti-apoptotic protein bcl-2 and decreasing the expression of pro-apoptotic proteins Caspase-3 and Bax.

BACKGROUND:It has been proved that miR-34a plays an inhibitory role in the growth of lung cancer stem cels, but the underlying mechanism remains unclear.
OBJECTIVE:To explore the inhibitory effect of miR-34a on lung cancer stem celsand the underlying mechanism.
METHODS:The CD133+lung cancer stem cels were separated from lung cancer A549 cel lines using magnetic activated cel sorting method. And miR-34a-overexpressing CD133+lung cancer stem cels were established by liposome transfection technology. Besides,the targeted relationship between miR-34a and Notch1 was analyzed by the dual-luciferase reporter. Afterwards, Notch1 silencing was performed by gene knockout, and its effect on lung cancer stem cels was determined.
RESULTS AND CONCLUSION:After sorted and detected by immunomagetic selection and flow cytometry assay respectively, a high rate of CD133+lung cancer stem cel was obtained. And qRT-PCR detected that the expression level of miR-34a in CD133+lung cancer stem cels was significantly lower than that in CD133-lung cancer stem cels. Moreover, miR-34a-overexpressing CD133+lung cancer stem cels were successfuly constructedandmiR-34a significantly inhibited proliferation and induced apoptosis of lung cancer stemcels. Dual-luciferase reporter assay indicated that Notch1 mRNA was a target of miR-34a. In addition, Notch1 silencing obviously inhibited proliferation and induced apoptosis of lung cancer stem cels. These findings suggest that miR-34a can inhibite lungcancer stem celsviathe Notch1 signaling pathway.

A new type of surface plasmon resonance ( SPR) spectroscopy system was designed and built. Here, a kind of dual photocell sensor was developed as a detection device to achieve a rapid measurement of SPR angle within a certain range. This SPR system was combined and integrated with electrochemical workstation to obtain a new type of electrochemistry-time-resolved SPR ( EC-TR-SPR ) instrument via instrumental technique. This EC-TR-SPR instrument was used to characterize the electrochemical polymerization process of aniline to validate the spectroscopic characteristics. Applications of transient electrochemical characterization methods, including chronoamperometry and differential pulse voltammetry, confirmed the time resolution and the applicability of this instrument system toward the steady state and transient electrochemical methods upon small molecular reactions. The experiment results showed that this EC-TR-SPR possessed the time resolution up to 10000 times per second (0. 1 ms), and could be used to real-time investigate the doping and de-doping of polymerization process of aniline monomer as well as the prepared polyaniline film, which could not be discriminated on a conventional electrochemical current-time curve. .