MiR-340 mediates the involvement of high mobility group box 1 in the pathogenesis of liver fibrosis / 中华肝脏病杂志

Objective: To explore the pathogenic mechanism of the miR-340/high mobility group box 1 (HMGB1) axis in the formation of liver fibrosis. Methods: A rat liver fibrosis model was established by injecting CCl(4) intraperitoneally. miRNAs targeting and validating HMGB1 were selected with gene microarrays after screening the differentially expressed miRNAs in rats with normal and hepatic fibrosis. The effect of miRNA expressional changes on HMGB1 levels was detected by qPCR. Dual luciferase gene reporter assays (LUC) was used to verify the targeting relationship between miR-340 and HMGB1. The proliferative activity of the hepatic stellate cell line HSC-T6 was detected by thiazolyl blue tetrazolium bromide (MTT) assay after co-transfection of miRNA mimics and HMGB1 overexpression vector, and the expression of extracellular matrix (ECM) proteins type I collagen and α-smooth muscle actin (SMA) was detected by western blot. Statistical analysis was performed by analysis of variance and the LSD-t test. Results: Hematoxylin-eosin and Masson staining results showed that the rat model of liver fibrosis was successfully established. Gene microarray analysis and bioinformatics prediction had detected eight miRNAs possibly targeting HMGB1, and animal model validation had detected miR-340. qPCR detection results showed that miR-340 had inhibited the expression of HMGB1, and a luciferase complementation assay suggested that miR-340 had targeted HMGB1. Functional experiments results showed that HMGB1 overexpression had enhanced cell proliferation activity and the expression of type I collagen and α-SMA, while miR-340 mimics had not only inhibited cell proliferation activity and the expression of HMGB1, type I collagen, and α-SMA, but also partially reversed the promoting effect of HMGB1 on cell proliferation and ECM synthesis. Conclusion: miR-340 targets HMGB1 to inhibit the proliferation and ECM deposition in hepatic stellate cells and plays a protective role during the process of liver fibrosis.

Central visual function and inner retinal structure in primary open-angle glaucoma / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

To investigate associations between central visual function and inner retinal structure in primary open-angle glaucoma (POAG). This study enrolled 78 POAG patients and 58 healthy controls. POAG was classified into early glaucoma and moderate to advanced glaucoma. The following tests were performed on all participants: isolated-check visual evoked potential (icVEP) testing, 24-2 standard automated perimetry (SAP), and Cirrus optical coherence tomography (OCT) examinations. Signal-to-noise ratio (SNR) measures obtained from icVEP responses to isolated checks presented at four depths of modulation (DOMs; 8%, 14%, 22%, and 32%) were explored. Mean macular sensitivity (mMS) was assessed by calculating the mean sensitivities of central 12 SAP points. Ganglion cell layer+ inner plexiform layer thickness (GCL+IPLT) and peripapillary retinal nerve fiber layer thickness (pRNFLT) were measured by OCT scanning. For each group of subjects, linear relationships among the following measures were analyzed: SNR, mMS, GCL+IPLT, and pRNFLT. SNR, mMS, GCL+IPLT, and pRNFLT were all more significantly decreased in glaucoma than in controls (P<0.001). A significant positive association was found between SNR at 14% DOM and GCL+IPLT at the inferior sector in early glaucoma (r=0.465, P=0.004). In moderate to advanced glaucoma, significant correlations were found between SNR at 32% DOM and mean GCL+IPLT (r=0.364, P=0.023), superior GCL+IPLT (r=0.358, P=0.025), and mean pRNFLT (r=0.396, P=0.025). In addition, in moderate to advanced glaucoma, there were significant correlations between mMS and all relevant measures of retinal thickness (r=0.330-0.663, P< 0.010). In early glaucoma, significant correlations were found between mean mMS and minimum GCL+IPLT (r=0.373, P=0.023), and between inferior mMS and superior GCL+IPLT (r=0.470, P=0.003). Linear models provided a good explanation for the relationship between SNR and inner retinal thickness (IRT), whereas nonlinear models better explained the relationship between mMS and IRT. In early glaucoma, both SNR and mMS were related moderately and significantly to IRT, whereas in moderate to advanced glaucoma, mMS was more strongly correlated with IRT than SNR.

Effect of suppressive oligodeoxynucleotides on the functional differentiation in CD4(+) Th1 lymphocytes in mice in vitro / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of suppressive oligodeoxynucleotides (Sup ODN) on the Th1 differentiation of CD4(+)T splenetic lymphocytes in mice.@*METHODS@#The splenetic lymphocytes of BALB/c mice were separated, and then CD4(+) cells were purified with immune magnetic CD4(+) microbeads (positive selection). The purification was examined by fluorescence-activated cell sorter. CD4(+) cells, anti-CD3epsilon, anti-CD28, IL-12 and Sup ODN or control oligodeoxynucleotides (Con ODN) were co-incubated for 72 h. IFN-gamma and IL-4 in the supernatant were detected using enzyme-linked immunosorbent assay. The expression of T-bet mRNA in CD4(+) cells was tested by reverse transcription polymerase chain reaction.@*RESULTS@#Sup ODN could significantly inhibit the release of INF-gamma and increase IL-4 production respectively (P<0.01). T-bet mRNA of CD4(+) lymphocytes was remarkably inhibited by Sup ODN as well (P<0.01).@*CONCLUSION@#In the presence of pro-Th1-cytokines, Sup ODN may affect the differentiation of CD4(+) T lymphocytes in vitro. Sup ODN can promote CD4(+) T cells to differentiate into Th2, and suppress them into Th1.

Serum level of HMGB1 in patients with hepatitis B and its clinical significance / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate whether there is a possible role of pro-inflammatory cytokine high mobility group box protein 1 (HMGB1) causing liver failure in severe hepatitis B patients.</p><p><b>METHODS</b>Serum HMGB1 levels of chronic hepatitis B (CHB) patients with different clinical conditions were measured and the correlations between HMGB1 and TBil or PTA were analyzed. (1) 54 chronic hepatitis B patients in different clinical conditions were enrolled in our study. Their serum TBil and PTA levels were detected by routine methods. (2) Their serum HMGB1 levels were also detected. 100 KD super-filtration columns were used to get rid of large proteins in the serum and 10 KD columns were used to condense the protein. Western blot was used to determine HMGB1 levels, and correlations between HMGB1 and TBil or PTA were analyzed.</p><p><b>RESULTS</b>The detection rates of serum HMGB1 were 100% (23/23), 90% (9/10), and 55% (6/11) in 23 patients with hepatic failure, 10 patients with chronic severe hepatitis B, and 11 patients with chronic moderate hepatitis B respectively. The concentration of serum HMGB1 levels in these three groups was (83.4+/-21.3), (78.1+/-19.5) and (60.3+/-14.3) microg/L respectively. Serum HMGB1 was not detected in normal healthy controls and hardly detected in convalescent and mild hepatitis patients. There were positive correlations between HMGB1 and TBil and negative correlations between HMGB1 and PTA.</p><p><b>CONCLUSION</b>HMGB1 levels in serum were closely associated with disease severity in chronic hepatitis B patients. HMGB1 may play a key role in the pathogenesis of chronic severe hepatitis B and liver failure.</p>