Objective: To investigate the effects of interleukin ( IL) -10 on the proliferation of HaCaT cells and CaCl2 induced expression of differentiation markers and its possible molecular mechanisms． Methods: HaCaT cells were treated with various concentrations of IL-10 (0，3，10，30 ng / ml) for different time (0，24，48，72 h) ，cell proliferation was measured using MTS，and cell cycle was determined by flow cytometry．HaCaT cells were pretreated with IL-10 (final concentration 10 ng / ml) for 1 h，then incubated with or without CaCl2 (final concentration 1. 2 mmol / L) for 24，48，72 h ，Western blot was performed to detect the effect of IL-10 on the expression of HaCaT keratinocyte differentiation markers．After pretreatment of HaCaT cells with PD98059，an inhibitor of mitogen-activated kinase-ERK1 /2，and LY294002，an inhibitor of phosphatidylinositol kinase-serine / threonine kinase (PI3K-AKT) ，the total RNA and proteins were extracted separately，real time quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to examine the influence of IL-10 on the expression of differentiation markers (Keratin1，Keratin5，Involucrin) . Results : MTS results revealed that IL-10 (30 ng / ml and lower doses) did not alter the proliferation of HaCaT cells in 72 h．Flow cytometry analysis demonstrated that IL-10 had no significant influence on cell cycle progression．The results of Western blot showed that IL-10 upregulated the expression of differentiation markers Involucrin，while there was no significant effect on Keratin1 and Keratin5 ．Mechanism research analysis demonstrated that IL-10 could activate ERK1 /2 and AKT ，increase their phosphorylation levels ; RT-qPCR and Western blot results showed that PD98059 and LY294002 partially blocked IL-10 induced Involucrin expression． Conclusion At a particular concentration range，IL-10 has little effect on HaCaT proliferation ，but it partially upregulates the expression of differentiation marker Involucrin via the MAPKs-ERK1 /2 and PI3K-AKT pathways.

Objective:To investigate the effect of HNRNPL protein on the proliferative ability of primary hepatocellular carcinoma cells and its potential mechanism.Methods:Online public database and real-time quantitative PCR were used to analyze the difference of HNRNPL expression between cancer and adjacent tissues. The effects of HNRNPL on HCC cell MHCC97H and HepG2 proliferation and MAPK pathway were investigated by Western blot, cell counting assay, colony formation assay and nude mouse transplantation tumor experiments.Results:The level of HNRNPL mRNA was validated to be higher in HCC tissue (2.76±0.37) than in normal tissue (1.00±0.14) with statistical difference ( t=3.93, P=0.002). Colony formation assay showed that the colony numbers of two MHCC97H knockdown groups (33.3±7.7) and (43.3±2.2) were lower than their control group (84.3±6.2), and two HepG2 knockdown groups (59.0±15.5) and (41.7±4.8) were lower than their control group (200.3±6.2) with statistical difference (both P<0.01). HNRNPL knockdown decreased the proliferation ability and activation level of MAPK pathway in HCC cells. Overexpression of oncogene c-RAF partially alleviated the anti-proliferation effect of HNRNPL knockdown and rescued the tumorigenic capacity. Conclusion:HNRNPL can promote hepatocellular carcinoma cell proliferation by activating MAPK signaling pathway.

Objective To analyze the frequency of interleukin (IL)-22+CD161+CD4+ T cells in the peripheral blood mononuclear cells (PBMCs) in rheumatoid arthritis (RA) patients compared with healthy control subjects and investigate the relationship of IL-22+CD4+CD161+ T lymphocyte frequency changes with RA disease activity.In addition to explore the pathogenesis of RA,and to look for new treatment targets for RA.Methods Twenty-one RA cases were included in the Department of Rheumatology of Tangshan Gongren Hospital from 2017 to 2018.Fourteen patients were female and 7 were male with the age ranged from 36 to 74 years old.The average age of this group of patients was (55±10) years,the average disease course was (60±50) months.All patients fulfilled the classification criteria of American College of Rheumatology [American College of Rheumatology (ACR)].Twenty-one subjects were enrolled as the control group,all of them came to Tangshan Gongren Hospital for regular health check-up.Fifteen subjects in the control group were female and 6 were male.Their age ranged between 40-78 years old with the average age of (55±9) years.IL-22+CD4+CD161+ T cells in PBMCs were detected by flow cytometry.The frequency variation of different CD4+CD161 + T was compared between case and control groups.The correlation was studied between the frequency and RA disease activity score (DAS28),tender joints number,swollen joints number,red blood cell sedimentation rate,high sensitive C reactive protein and white blood cell counts,red blood cell counts,platelet counts,IgG,IgA,IgM,complement C3 level,complement C4 level.T-test or Mann-Whitney U test were used for single-factor analysis,Pearson's test was used for correlation analysis.Results The percentage of RA group secreted CD4+ T cells (0.33± 0.20)％ of INF-γand IL-22,CD4+ T cells (0.51±0.29)％ of IL-22,and CD4+CD161+ T cells of IL-22 simultaneously.The number (0.55 ±0.28)％ was.significantly higher than that of the healtby control group [(0.22±0.14)％,(0.25±0.18)％,(0.36±0.24)％],and the differences were statistically significant [P=0.002,P=-0.0.45,P=0.026].Conclusion The percentage of IL-22+CD4+CD161+ T lymphocytes in the peripheral blood monocytes in RA patients is significantly higher than that in the healthy controls.The results of this study suggest that IL-22+CD4+CD161+ T lymphocytes in RA patients maybe related to RA disease activity and joint lesions.

Objective To explore the application value of 3.0T MR diffusion tensor imaging (DTI)in 3D reconstruction of the knee peripheral nerve.Methods DTI with 25 motion probing gradients (MPGs)was performed on the left knee in 22 healthy young volunteers;By choosing the original images scaned in the vertical direction,three post-processing methods including maximum intensity proj ection (MIP),volume rendering (VR)and diffusion tensor tractography (DTT)were used.Two radiologists evaluated these images visually.For the same nerve,MIP,VR and DTT were compared using Friedman test.Intraclass correlation coefficient (ICC )analysis was used to compare the correlation among the three imaging methods.Score consistency between two observers was calculated by using the Kappa statistic.Results For the same nerve,there was no significant statistical difference for the MIP,VR and DTT imaging quality scores(P>0.05).The I CC value and the k value were 0.718-0.856,0.668-0.705,respectively.Conclusion Stereoscopic and clear displaying the tibial nerve/the common peroneal nerve in the knee can be achieved by using MIP,VR and DTT post-processing methods based on DTI original images.The reconstruction of MIP,VR is simple and efficient,in which the objectivity is better than DTT.The reconstruction of MIP,VR can be used as effective preview methods before the DTT reconstruction with complexity and strong subjective dependence.

Objective To study the morphology and characteristics of the popliteal tibial nerve and measure relevant structural parameters of the peroneal nerve passing round the fibular head and neck by 3D(three-dimensional)MR advantage sequences,and provide valuable image information for basic research and clinical work.Methods Thirty-six left knee joints of healthy volunteers were scanned using 3D MR sequences,including 3D-FS-CUBE,3D-FS-SPGR.The anatomical features in MR imaging were observed by two doctors.The different level axis section areas of the popliteal tibial nerve were measured by reconstructed 3D MR advantage sequences.The coronal/sagittal diameters and the angle of the fibular head and neck(peroneal tunnel)were also measured and compared.SPSS 19.0 software was used for further statistical analysis.Results Optimum visual rates of the knee nerve of 3D-FS-CUBE,3D-FS-SPGR sequences were 25%,89%(P<0.01).Axial section of the tibial nerve of the popliteal space was as following:top level(Tn1,tibial nerve)(19.3 ± 1.02)mm2,midpoint level(Tn2)(23.6 ± 2.24)mm2,supracondylar level(Tn3)(29.1 ± 3.39)mm2.Coronal and sagittal diameters of the fibular head were(19.8 ± 2.95)mm,(19.7 ± 4.2)mm,and coronal and sagittal diameters of the fibular neck were(12.7 ± 3.1)mm, (13.6 ± 2.5)mm,respectively.The angle between the head and neck was 164°± 6.5°.Conclusion 3D-FS-SPGR sequence is the advantage sequence for displaying the knee nerves.The diameters of the popliteal tibial nerves gradually thicken from Tn 1 to Tn3.The structural measurements of the fibula head and neck have certain significance for fundamental research or prevention and treatment of common peroneal nerve entrapment.

Alzheimer's disease ( AD ) is a complex disease caused by environmental and genetic factors. Therefore, one-drug-multiple-target compounds represent the most promising pharmacological approaches to preventing and treating this dis-ease. We have previously designed and synthesized bis( n)-Cog-nitin, novel anti-Alzheimer's dimers derived from tacrine. Bis ( n)-Cognitin have been proven to act on multiple important AD targets, including acetylcholinesterase, β-secretase, N-methyl-D-aspartic acid receptor and neuronal nitric oxide synthase, con-currently. Moreover, Bis(n)-Cognitin could inhibit β-amyloid-induced neurotoxicity, decrease glutamate-induced excitotoxici-ty, reduce oxidative stress, improve learning and memory, and protect against neuronal apoptosis in various in vitro and in vivo models, suggesting that bis ( n )-Cognitin are potential anti-AD drug candidates.

Objective To investigate the effect and mechanism of decitabine (DAC) on the proliferation of human cholangiocarcinoma CCLP1 cells in vitro and in vivo.Methods After treated with various concentrations of DAC,cell growth inhibition rates were determined by CCK-8 assay.Cell cycle and cell apoptosis were analyzed by flow cytometry.Cell autophagy was observed under fluorescence microscope.The effect of DAC on the growth of cholangiocarcinoma in vivo was determined in a CCLP1 mice xenograft model.Results The proliferation rate of CCLP1 cells in the DAC-treated group decreased in a time-concentrated dependent manner.After treatment with DAC,the cell cycle of CCLP1 cells was arrested at the G2/M phase.The apoptosis rate was significantly higher in the treatment group over the control group.Cell autophagy was observed after treatment with DAC in CCLP1 cells.The tumor growth of implanted CCLP1 cells significantly slowed down after the mice were treated with 0.8 mg/kg DAC,6 times weekly for 2 weeks.Conclusion DAC can inhibit the proliferation of cholangiocarcinoma in vitro and in vivo.

Objective To control IDS virus (HIV)transmission through blood, provide low -risk blood donors to donate regularly, and ensure blood safety and reduce the risk of blood transfusion. Methods By analyzing HIV antibody(anti-HIV) test results, HIV infection and prevalence of blood donors from 2000 to 2009, susceptible people were identified,prevented,educated,in order to prevent the spread of HIV. Results A total of 10 years in Huainan region 361 cases of anti-HIV positive were screened out. Anti-HIV-positive of voltmtary blood donors were recognized anti-HIV positive in 4 cases ,accounting for 0.209/10 000 of the total voluntary blood donation. Epidemiological analysis in 4 cases of anti-HIV confirmed positive results: the education level of all was less than high school,75% were the local household registration ;25% were mobile population, and all were young adults, aged from 23 to 38 years;75% were workers,and 25% were temporary workers and the service sector. Conclusion By strictly blood screening to blood donation, strengthening the management, intensify propaganda against AIDS in efforts, raising people 's awareness of prevention,the spread of AIDS could be prevented and controlled.

Purpose To investigate the effect of interleukin-10(IL-10)on IL-15mRNA and IL-6mRNA in Hela cells induced by lipopolysaccharide(LPS)and to analyze their activiated signal transduction pathways.Methods Extracted total RNA and total proteins of cultured Hela cells,which were treated with different concentrations of LPS,or IL-10 alone or in combined use,were to analyze the levels of IL-15mRNA and IL-6mRNA using RT-PCR and to analyze the expression of signal transduction pathway proteins using Western blot.Results RT-PCR indicated that expression of IL-15mRNA and IL-6mRNA in Hela cells strikingly increased after 12 h using 1 ng-10μg LPS(P＜0.01 vs control),which had dose-dependence and achieved peak value using 100 ng/mL.During 0-24 h,expression of IL-15mRNA and IL-6mRNA strikingly increased with time changing(P＜0.01 vs control),which had time-dependence and achieved peak value at 12 h.Expression of IL-15mRNA and IL-6mRNA had no conspicuous difference in Hela cells treated with IL-10(10 ng/mL)alone(P＞0.05 vs control).Different concentrations of IL-10 1,10,100 ns/mL)markedly downregnlated expression of IL-15mRNA and IL-6mRNA in Hela cells induced by LPS.Furthermore,the effect of inhibition will be more obvious with dose increasing.Western blot indicated that LPS upregulated expression of IL-15mRNA and IL-6mRNA by phosphorylation of PI3K/AKT and ERK1/2.IL-10 blocked phosphorylation of AKT,but could not affect phosphorylation of ERK1/2.Conclusion IL-10 downregulated the expression of inflammatory cytokines IL-15 and 1L-6 induced by LPS,which may correlate with the fact that phosphorylation of AKT was blcoked by IL-10.Therefore,IL-10 may be used to prevent and treat some clincal infective diseases.

Purpose To explore the antiviral effect of interleukin 29(IL-29) on hepatitis B virus in vitro.Methods To study the antiviral effect of IL-29 against hepatitis B virus by the amount of HBV mRNA detected .Through the quantity of mRNA translated from genes of MxA,2′,5′-OAS,PKR and RNase L as well as the signal pathway induced by IL-29,we used RT-PCR and Western blot to discuss the anti-hepatitis B virus mechanism which was stimulated by IL-29.Results The amount of HBV mRNA in HepG2.2.15 cells was reduced by stimulation of IL-29.The expression of MxA and 2′,5′-OAS was up-regulated,as well as P-ERK and P-AKT were activated by IL-29.Conclusion These findings showed that IL-29 had obvious antiviral activity towards HBV in HepG2.2.15 cells.