The expression of microRNA (miRNA) is closely related to radio-chemosensitivity in glioma stem cells (GSCs).Moreover,the growth of glioma stem cells could be inhibited comprehensively by increasing radio-chemosensitivity and apoptosis,simultaneously with the regulation of a single miRNA,which has been confirmed by some researches.Thereby microRNA is prospective for the adoption as a specific agent in targeted therapy of glioma,so as to increase the radio-chemosensitivity in glioma stem cells.

OBJECTIVE:To investigate the effect of aqueous extraction of Lonicerae Flos (SYHW) on anti-influenza A virus H1N1 (H1N1 virus) in vitro. METHODS:Using Madin-Darby canine kid ney (MDCK) cells cultured in vitro by H1N1 virus, half of the tissue culture infection dose(TCID50)was calculated. Culturing MDCK cells for 24 h with different mass concentrations of SYHW,the maximum non-toxic concentration was investigated. And then test was divided into normal cell group,virus control group,SYHW preventive administration group,therapeutic administration group and direct killing group (given SYHW of maxi-mum non-toxic concentration,infecting cells by 100 TCID50 H1N1 virus),and antiviral effective rate (ER) of SYHW was deter-mined. Test was divided into normal cell group,virus control group,SYHW therapeutic group and direct killing group (the same administration and infection as above),changes of cell proliferation index (PI) and cell apoptosis rate were respectively deter-mined. RESULTS:100 TCID50 of H1N1 virus was 1.26×10-7,and the maximum non-toxic concentration of SYHW on MDCK cells was 50 μg/mL(cell survival rate was 91.3%). ERs of preventive administration group,therapeutic administration group and direct killing group were 0,80.3% and 52.7%,respectively. Compared with normal cell group,PI value in virus control group was sig-nificantly reduced (P<0.05),early and late apoptotic rates were significantly increased (P<0.05). Compared with virus control group,PI value in directly killing group was significantly increased(P<0.05),and early apoptotic rate was significantly reduced (P<0.05);early apoptotic rates in therapeutic administration group were significantly reduced (P<0.05). CONCLUSIONS:SY-HW shows anti-H1N1 virus effect in vitro,therapeutic administration and directly killing are preferred in antiviral effect.

Objective To investigate the mechanism of Roscovitine, an inhibitor of cyclin-dependent kinase (CDK), in inhibiting HBV replication.Methods Recombiant expression plasmids of SAMHD1 (sterile alpha motif and histidine/aspartic acid domain-containing protein 1) mutants that were defective in dNTPase (deoxynucleoside triphosphate triphosphohydrolase) activity and phosphorylation at the threonine (T) 592 residue were constructed.Huh7.0 cells were respectively co-transfected with different SAMHD1 mutants in combiantion with HBV replication plasmid to analyze whether the retroviral restriction ability of SAMHD1 was regulated by phosphorylation.The cytotoxicity of Roscovitine to Huh7 cells was evaluated by MTT assay.HBV core-associated DNA levels and phosphorylation of SAMHD1 in transfected Huh7.0 cells which were treated with different concentrations of Roscovitine were measured by Southern blot and Western blot assays.Results The SAMHD1 mutant that was defective in the dNTPase active site of D207N lost its ability to restrict HBV replication.Dephosphorylation of SAMHD1 at T592 enhanced its restriction on HBV.The median toxic concentration (TC50) of Roscovitine was 11.20 μmol/L.Both the HBV core-associated DNA levels and the phosphorylation of SAMHD1 were down-regulated by Roscovitine.Conclusion The anti-HBV function of SAMHD1 in dividing cells is regulated by phosphorylation.Roscovitine can inhibit the replication of HBV through reducing the phosphorylation of SAMHD1.

Objective:To study the role and mechanism of heat shock protein 70 (HSP70) in apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3B (APOBEC3B)-mediated inhibition of hepatitis B virus(HBV) replication.Methods:The interaction between HSP70 and APOBEC3B was analyzed by co-immunopreciptation (Co-IP) and GST pull-down. After treating Huh7 cells with siHSP70 or HSP70 inhibitor VER155008 or overexpressing HSP70 in Huh7 cells, changes in the antiviral effect of APOBEC3B were detected by Southern blot and real-time PCR; the deaminase activity of APOBEC3B was tested by differential DNA denaturation PCR(3D-PCR) and clone sequencing.Results:HSP70 could bind to APOBEC3B. Overexpression of HSP70 promoted the deaminase activity and anti-HBV activity of APOBEC3B. On the contrary, knockdown of HSP70 or using HSP70 inhibitor VER155008 would attenuate the deaminase activity and anti-HBV activity of APOBEC3B.Conclusions:HSP70 could promote the anti-HBV activity of APOBEC3B by enhancing the deaminase activity of APOBEC3B.

Objective To study the calcium metabolism in tacrolimus(FK506)induced rats nephrotoxicity and the preventive effect of calcium channel blocker.Methods Twenty-four Spragueinduced or FK506-induced nephropathy model.Blood creatinine,blood electrolytes,renal tissue histopathology(HE stain)and the change of ultrastructural organization in renal cells by transmission electron microscope were observed.Results The blood creatinine levels of both CsA and FK506 groups [(36.00±2.61)and(34.17±4.54)μmol/L] were significantly higher than those of the FK506+Dilgroup and control group(all P＜0.05).The blood calcium levels of both CsA and FK506 groups (2.00±0.04 and 2.05±0.04 mmol/L) were significantly lower than those of the FK506+Dil group and control group(all P＜0.05).The blood creatinine and calcium levels of FK506+Dil group were not significantly different with those of control group(P＞O.05).Histopathology examination showed cloudy swelling and vacuolization of the renal tubular epithelial cells and intra-cellular mitochondria swelling and vacuolization in the CsA and FK506 groups.However,the pathological changes of the FK506+Dil group were remarkably milder in comparison with the CsA and FK506 groups.Concluum channel blocker,Dil,could prevent the FK506-induced nephrotoxicity.

We proposed a new stitching method based on sift features to obtain an enlarged view of transmission electron microscopic (TEM) images with a high resolution. The sift features were extracted from the images, which were then combined with fitted polynomial correction field to correct the images, followed by image alignment based on the sift features. The image seams at the junction were finally removed by Poisson image editing to achieve seamless stitching, which was validated on 60 local glomerular TEM images with an image alignment error of 62.5 to 187.5 nm. Compared with 3 other stitching methods, the proposed method could effectively reduce image deformation and avoid artifacts to facilitate renal biopsy pathological diagnosis.
Algorithms ; Artifacts ; Humans ; Image Processing, Computer-Assisted ; methods ; Kidney Glomerulus ; ultrastructure ; Microscopy, Electron, Transmission ; methods

Objective To investigate the regulatory role of host restriction factor Moloney leukemia virus 10 (MOV10) protein in HBV replication. Methods Firstly, a HBV replication-expression plasmid was transfected into Huh7 cells to investigate the effect of HBV replication on MOV10 expression. Secondly, HBV DNA was extracted and measured by quantitative PCR ( qPCR) after knocking down the expression of endogenous MOV10 or enhancing the expression of exogenous MOV10. Furthermore, MOV10 conserved do-mainⅡenzyme active mutants (D645A, E646Q and G648A) were constructed and analyzed regarding their antiviral activities. The HBV replication plasmid and MOV10 expression plasmid were co-transfected into hu-man renal epithelial cells (HEK293) to investigate whether MOV10 could bind to HBV mRNA using RNA binding protein immunoprecipitation ( RIP) . Results The expression of MOV10 was increased after trans-fection of HBV replication plasmid into Huh7 cells. After knocking down the expression of endogenous MOV10 by siRNA in Huh7 cells, HBV replication was increased about 1. 5 times compared with control group, while the viral DNA level was significantly decreased in Huh7 cells that overexpressed MOV10. MOV10 domain Ⅱ mutants also significantly inhibited HBV replication. MOV10 could bind to 3. 5 kb HBV RNA. Conclusion In liver cancer cells, the expression of the host restriction factor MOV10 was associated with HBV replication. Its inhibitory effect against HBV replication was independent of its helicase activity, but might be associated with its binding activity with 3. 5kb HBV RNA.

Chronic hepatitis B virus (HBV) infection is one of the leading causes of hepatocellular carcinoma (HCC). Ubiquitin-proteasome system (UPS) mediates the post-translational modification and degradation of protein in eukaryotic cells. Recent studies have revealed that UPS plays an important role in HBV infection and hepatocarcinogenesis. Targeting HBx to intervene in the progression of HBV infection or HBV-related HCC has become a research hotspot both domestically and internationally in recent years. This article reviewed the progress in HBx promoting HBV infection and hepatocarcinogenesis through UPS.

Objective To investigate the potential of polylactic acid glycolic acid copolymer (PLGA) composite ordered multi tunnel collagen scaffold in fabricating a biomimetic artificial nerve graft to repair the sciatic nerve defects in rats.Methods The ordered multi tunnel collagen scaffold was prepared by vacuum freeze-drying and directional drawing method to simulate the epineurium;the outer conduit was prepared by PLGA to simulate the epineurium;and then,the ordered multi tunnel collagen scaffolds were loaded in the PLGA conduit (5∶1) under a stereomicroscope to develop a novel biomimetic artificial nerve.Sixty-four rats were randomly divided into four groups:artificial nerve group,PLGA group,peripheral nerve group,and non-graft control group (n=16);rats in the artificial nerve group,PLGA group,and peripheral nerve group were repaired with artificial nerve graft,hollow PLGA conduit and allogeneic sciatic nerve to bridge the sciatic nerve defect,while the sciatic nerve with the gap in rats of the control group was without any grafting.After 11 weeks of operation,the hind limbs of rats in each group were detected by behavioral test,electrophysiological examination and Fluoro-Gold retrograde tracing method.The changes of muscle tissues (gastrocnemius) were observed by hematoxylin staining and TMR-α-BTX staining,and the regenerated axons were observed by immunohistochemical staining with NF200 and the regenerative spinal anterior horn motor neurons were observed by Nissl fluorescence staining 12 weeks after operation.Results After 11 weeks of operation,the recoveries of the motor functions (the distance between the injured hindlimb and forelimb,the rotation angle of the injured foot) in the peripheral nerve group,artificial nerve group,PLGA group and control group were significantly deteriorated in turn,and the differences were statistically significant (P＜0.05).Electrophysiological examination showed that the recovery effect of peripheral nerve group was the best in both latency and amplitude of the compound muscle action potential,followed by artificial nerve group.The latency of PLGA group was the longest and the amplitude of compound action potential was the smallest;significant differences were noted between each two groups (P2＜0.05).At 12 weeks after operation,the wet weight ratio of muscle fibers,area of muscle fibers and neuromuscular junction area were significantly different between each two groups (P＜0.05);the degree of gastrocnemius atrophy in the artificial nerve group was significantly improved than that in the PLGA group,but not yet reached the level of peripheral nerve group.NF200 immunohistochemical staining showed that a large number of NF200-positive axons were seen in the grafts of the artificial nerve group,but the number was slightly smaller than that of the peripheral nerve group;the number of regenerated axons in the PLGA group was the smaller and mainly distributed near the proximal side.In the PLGA group,only (19.33 ±6.73)％ regenerated spinal anterior horn motor neurons were labeled with Fluoro-Gold,while the positive rates of Fluoro-Gold in the artificial nerve group and peripheral nerve group were (42.67±7.45)％ and (50.13±4.33)％;the differences between each two groups were statistically significant (P＜0.05).Conclusion The biomimetic artificial nerve made of PLGA conduit and ordered multi tunnel collagen scaffold can efficiently reconstruct the defected peripheral nerve with guiding axonal regeneration and promoting functional restoration in rats;however,its effect is poor than peripheral nerve grafting.

The cross regional loose medical alliance is an important carrier in the current integrated development process of medical services in the Yangtze River Delta region. Smith policy implementation process model was used to analyze the development difficulties of cross regional loose medical alliances from idealized policies, policy implementation institutions, policy target groups, and policy implementation environment. Such medical alliances were formed under the background of integrated development in the Yangtze River Delta, with Shanghai′s tertiary public hospitals as leading units and medical institutions in Jiangsu, Zhejiang, and Anhui provinces as member units. Analysis showed that the policies for such medical alliances development had not yet clearly defined the organizational management mode, operational mechanism, and implementation path, and the corporate governance structure of medical alliance was immature; The policy implementation agencies were relatively lagging behind in the support of special funds and the formulation of related supporting policies; Participation of policy target groups was insufficient and their incentive mechanisms was imperfect; There were problems in the policy implementation environment, namely inconsistent medical and health service regulations and systems in different regions, different health financing capabilities of local governments, insufficient coordination of medical institution management concepts, and a lack of unified standards in information systems. Based on the above difficulties, this study proposed to strengthen the development planning and layout of cross regional loose medical alliances, and improve the corporate governance structure; To strengthen the government′s main responsibility and improving policy implementation capabilities; To improve the internal cooperation and operation mechanism of cross regional loose medical alliances, and enhance the sense of identity of the target group; To optimize the policy implementation environment and implement various support measures, so as to provide references for further promoting the coordinated development of high-quality medical resources in the Yangtze River Delta region.