OBJECTIVES: To investigate the expression of the long non-coding RNA maternally expressed gene 3 (LncRNA Meg3) in patients with the Wilson disease (WD) and its correlation with the severity of liver fibrosis and autophagy-related markers. METHODS: A total of 100 WD patients and 50 healthy individuals were enrolled from the First Affiliated Hospital of Anhui University of Chinese Medicine. Serum biomarkers, including platelet count, hyaluronic acid (HA), laminin (LN), type III procollagen N-terminal peptide (PIIINP), type IV collagen (C‑IV), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), were measured, and the non-invasive indices APRI and FIB-4 were calculated. Peripheral blood levels of LncRNA Meg3, Beclin-1 and LC3B were detected using RT-qPCR, and liver stiffness (LSM) and shear wave velocity (SWV) were evaluated using two-dimensional shear wave elastography (2D-SWE). The liver tissues from 10 WD patients and 10 patients with hepatic hemangioma were examined using histochemical staining, transmission electron microscopy, and RT-qPCR. RESULTS: The expression level of LncRNA Meg3 was significantly lower, while the levels of AST, ALT, HA, LN, PIIINP, C‑IV, APRI, FIB-4, LSM and SWV were significantly higher in WD patients than in the healthy individuals (all P<0.01). LncRNA Meg3 was negatively correlated with LSM, SWV, APRI, FIB-4, Beclin-1 and LC3B (P<0.05). ROC analysis demonstrated that LncRNA Meg3 effectively discriminated >F4 stage fibrosis (AUC=0.902) with a sensitivity of 92.9% and a specificity of 83.7% at the optimal cut-off value, outperforming APRI (AUC=0.746) and FIB-4 (AUC=0.661). The liver tissues from WD patients exhibited characteristic histopathological changes and lowered expression of LncRNA Meg3, which was negatively correlated with Beclin-1 and LC3B expressions (P<0.05). Liver fibrosis staging (7 S4 cases and 3 S3 cases) was significantly associated with LSM and SWV levels (P<0.05). CONCLUSIONS The expression level of LncRNA Meg3 is significantly decreased in WD patients, which is negatively correlated with the severity of liver fibrosis and closely related to the level of autophagy.
Humans ; RNA, Long Noncoding/metabolism* ; Liver Cirrhosis/metabolism* ; Adult ; Female ; Male ; Hepatolenticular Degeneration/metabolism* ; Case-Control Studies ; Young Adult ; Adolescent ; Middle Aged

BACKGROUNDThe prevalence of metabolic syndrome (MetS) increased recently and there was still not a screening index to predict MetS. The aim of this study was to estimate whether brachial-ankle pulse wave velocity (baPWV), a novel marker for systemic arterial stiffness, could predict MetS in Chinese community population.

METHODSA total of 2 191 participants were recruited and underwent medical examination including 1 455 men and 756 women from June 2011 to January 2012. MetS was diagnosed according to the criteria of the International Diabetes Federation (IDF). Multiple Logistic regressions were conducted to explore the risk factors of MetS. Receiver operating characteristic (ROC) curve was performed to estimate the ideal diagnostic cutoff point of baPWV to predict MetS.

RESULTSThe mean age was (45.35±8.27) years old. In multiple Logistic regression analysis, the gender, baPWV and smoking status were risk factors to MetS after adjusting age, gender, baPWV, walk time and sleeping time. The prevalence of MetS was 17.48% in 30-year age population in Shanghai. There were significant differences (χ(2) = 96.46, P < 0.05) between male and female participants on MetS prevalence. According to the ROC analyses, the ideal cutoff point of baPWV was 1 358.50 cm/s (AUC = 60.20%) to predict MetS among male group and 1 350.00 cm/s (AUC = 70.90%) among female group.

CONCLUSIONBaPWV may be considered as a screening marker to predict MetS in community Chinese population and the diagnostic value of 1 350.00 cm/s was more significant for the female group.

Adult ; Ankle Brachial Index ; methods ; Female ; Humans ; Logistic Models ; Male ; Metabolic Syndrome ; diagnosis ; Middle Aged ; ROC Curve

Objective To investigate the pathogenesis of cytotoxic T cell (CTL) dysfunction in patients with HCV infection. Methods CTL detecting system was established. Two polypeptides which could enhance CTL function and two polypeptides which could inhibit CTL function were selected and cross-combined. BALB/c mice were immunized by subcutaneous injection of the combined polypeptides, and the CTL activity in mouse spleen cells was detected by LDH release test. Results CTL activity in BLAB/c mice immunized by polypeptides in the core region of HCV could be enhanced by CPA10 (5-23 aa) and inhibited by CPA9 (39-74 aa). CTL activity in the mice could be enhanced by polypeptides from the HCV core region, CPB2+CPB8, and CPB6+CPB8, respectively. There was no obvious difference between CPB2+CPB7, CPB6+CPB7 and the negative control. Two-factor analysis of variance showed that there was reciprocal action between the inhibitory and enhancing polypeptides from the HCV core region. Conclusion CTL activity in BLAB/c mice can be detected stably by LDH. There is an interactive effect between the inhibitory and enhancing polypeptides from the HCV core region.

OBJECTIVETo investigate the pathogenesis of cytotoxic T cell (CTL) dysfunction in patients with HCV infection.

METHODSBALB/c mice were immunized by subcutaneous injection of polypeptides from HCV core region, and the CTL activity of mouse spleen cells was detected by the LDH release test. Two polypeptides which can enhance CTL function and two polypeptides which can inhibit CTL function were selected and cross-combined. BALB/c mice were immunized using the combined polypeptides and the CTL activities were detected afterwards.

RESULTSCTL activity was inhibited by CPA9 (39-74 amino acids), CPB7 (67-76 amino acids) and CPB8 (71-80 amino acids), and promoted by CPA10 (5-23 amino acids), CPB6 (63-72 amino acids) and CPB2 (131-140 amino acids). Using single factor analysis of variance, the CTL activity in the mice could be enhanced by polypeptides from the HCV core region, CPB2+CPB8, CPB6+CPB8, respectively. There was no obvious difference between CPB2+CPB7, CPB6+CPB7 and negative control.

CONCLUSIONSCPA9, CPB7, and CPB8, the 3 polypeptides from HCV core region play an inhibition role and CPA10, CPB6, and CPB2 play an enhancement role in CTL activity in mice. The inhibition and enhancement functions of the polypeptides from HCV core region interact each other.

Animals ; Cytotoxicity Tests, Immunologic ; Cytotoxicity, Immunologic ; drug effects ; Mice ; Mice, Inbred BALB C ; Peptide Fragments ; administration & dosage ; immunology ; Spleen ; cytology ; drug effects ; immunology ; T-Lymphocytes, Cytotoxic ; cytology ; immunology ; Viral Core Proteins ; administration & dosage ; chemistry ; immunology

Objective　To study the similarities and differences on in vitro replication and expression of hepatitis C virus (HCV) between human fetal hepatocytes (HFH) and 7721 cell line. Methods　Human fetal hepatocytes and a hepatoma cell line 7721 were incubated with a serum from hepatitis C patient. After incubation, the presence of HCV RNA, the expression of HCV NS3 antigens in cells and/or supernatant were examined by RT-PCR, in situ hybridization and immunohistochemistry, respectively. Results　It was found that: ①The intracellular HCV RNA was first detected on d 2～3 post-incubation and then could be intermittently detected in cells and/or supernatant subsequently (HCV RNA could be detected in 7721 cells during a period of at least 66 days. In HFH, HCV RNA could be detected up to 25 days after incubation); ②HCV-NS3 antigen could be expressed in infected cells; ③Minus-strand RNA of HCV was mainly located within cytoplasm by in situ hybridization. Conclusion　The results suggest that both the fetal hpatocytes and the hepatoma cell line 7721 are susceptible to HCV, and especially 7721 cell line can stably support HCV replication in vitro and may be used as the target cell for long-term cultures of HCV.

Objective To explore the mechanism of CTL dysfunction in HCV infected person, to provide a theoretical basis for further understanding the pathogenesis of hepatitis C and the development of HCV vaccines. Methods HCV nucleopolypeptides were selected and synthesized with the method of solid phase synthesis. BALB/c mice were immunized subcutaneously with HCV nucleopolypeptides, and CTL activity of mice was detected by LDH releasing test. Results CTL of mice could be inhibited by HCV nucleopolypeptides residues 39-74, 67-76, 71-80 and enhanced by HCV nucleopolypeptides residues 5 23,63 72,131 140. Conclusion The function of CTL can be suppressed and intensified by different HCV nucleopolypeptides.

Objective To establish a simple animal model and the cytotoxic T cell(CTL) detecting system for the studies of the effects of CTL on HCV infection. Methods The CTL activity in BLAB/c mice immunized by polypeptides in the core region of HCV was detected with lactic dehydrogenase (LDH) by using SP2/O cells as the target cells. Results CTL activity in BLAB/c mice immunized by polypeptides in the core region of HCV could be detected with LDH. The activity could be enhanced by CPA10(5～23 aa) but inhibited by CPA9(39～74 aa). Conclusion CTL activity in BLAB/c mice can be detected stably by LDH.