Objective To study the growth effect of human cholangiocarcinoma cell line QBC939 treated by norcantharidin (NCTD) and preliminary illustrate the potential mechanism.Methods The human cholangiocarcinoma cell line QBC939 was detected by MTT assay,flow cytometry,immunocytochemistry after the treatment of NCTD in vitro.Results NCTD displayed inhibitory effect on growth of QBC939 from different doses of 0.125,0.75,2.5,10,120 μg/ml after 48 h (P ＜0.05).It was in a dose and time dependent manner.Dose-effect curve was drawn and IC50 value was (3.66±1.14) μg/ml.The flow cytometric profiles showed that the rate of cell apoptosis enhanced following increasing the concentration of NCTD[(8.6±0.4) ％,(17.6±0.3) ％,(22.9±0.4) ％,(25.5±0.9) ％ and (31.1±1.5) ％,respectively]and cells blocked in the G2/M phase after treatment with 2.5 μg/ml NCTD[(14.1±1.0) ％ and (5.7±0.3) ％].The expression of the protein caspase-3 elevated after different concentrations of NCTD co-cultured with QBC939 compare with contrast group.Conclusion NCTD has an inhibitory effect on proliferation of QBC939 cell line,and the mechanism might be related to the induction of cell apoptosis and blockade of cell cycle.

Objective:To better understand the changes of the NKT like cells after HIV infection and HAART treatment.Methods: Peripheral blood from HIV-infected individuals, HAART-treatment AIDS patients and healthy controls were collected, the expression of CD57 and the proliferation ability of NKT like cells before and after HAART were analyzed by flow cytometry.Results:We found that the percentage of NKT like cells before HAART was significantly lower than the healthy controls ( P<0.01 ) , and recovered after HAART treatment ( P<0.05 );the aging of NKT like cells was significantly higher before HAART compared with health individuals (P<0.01),and recovered after HAART treatment(P<0.05)the proliferation was significantly lower in vitro before HAART compared with healthy controls,and partial recovered after HAART.Conclusion: After HAART treatment,the number of NKT like cells,CD57 expression and the proliferation ability of HIV infected patients were restored.

Objective:To study the changes of the NKT like cells after HIV infected. Methods:We collected peripheral blood from 47 untreated HIV infected individuals and 31 healthy controls,and analyzed the expression of Annexin-V,Ki-67,HLA-DR and other surface molecules in NKT like cells by flow cytometry. Results:The NKT like cell percentage of untreated HIV infected group was (3. 03±1. 61)%,the NKT like cell percentage of normal control group was (8. 30±7. 42)%,the percentage of NKT like cells in HIV infected individuals was significantly lower than the healthy controls ( P<0. 05 );the NKT like cell HLA-DR expression of untreated HIV infected group and normal control group were (5. 40±4. 10)% and (0. 89±0. 83)%,the NKT like cell Annexin-V expression of untreated HIV infected group and normal control group were (30. 21±13. 15)% and (5. 40±8. 05)% ,and the activation and apoptosis of NKT like cells was significantly higher after HIV infection compared with health individuals (P<0. 001,P<0. 01),the degree of activation was negatively correlated with CD4 count (r=-0. 885 7,P<0. 05);and the NKT like cell Ki-67 expression of untreated HIV infected group and normal control group were (11. 15±4. 76)% and (27. 63±18. 31)%,the proliferation ability was significantly lower after HIV infection compared with healthy controls(P<0. 05). Conclusion:HIV infection can significantly reduce the number of NKT like cells and its ability to proliferate,and increase its ability to activation and apoptosis.

ObjectiveTo screen mimetic HIV-1 neutralizing epitopes from plasma with high level neutralizing antibody,and to provide useful information for further study of the interaction between antigen and antibody.MethodsIn order to gain neutralizing antibody recognized mimotopes, we detected neutralizing antibodieslevelsof 11HIV-1infectedslowprogressorsbyPBMC-basedneutralization assays.High-titer HIV-neutralizing antibodies from plasma of SPs was used as the ligand for biopanning by phage-displayed random peptide library.Positive phage clones was evaluated by ELISA,sequenced,and analyzed for homology to HIV-1 env by local BLAST to deduce the neutralizing peptide.ResultsTwenty-two clones were obtained consistent with requirement through three rounds biopanning.After comparison analysis,twelve clones include C8 were obtained as mimotopes of neutralizing antibody,C40 located in gp41Ⅱ cluster with the highest titer by inhibition ratio may be as neutralizing epitope.Conclusion By the use of IgG antibodies from SPs to screen the phage random polypeptide library,one can acquire multiple phage mimetic peptides of HIV related antigen epitope.(Chin J Lab Med,2012,35:838-842 )

Objective To discusse the research status ofyanjing medical school experience method, and provide a reference for relative researchers.Methods Articles in TCM Database System, SinoMed and CNKI were retrieved refined and analyzed t from the year, author, journal, organization and so on after duplicate removing.Results There were 389 related articles published in journals from 1956 to 2012. With 41.90% articles published in 10 core journals, written by 14 main authors. There were 3 hot research areas including experience of famous doctor,syndrome differentiation and treatment and medical record.Conclusion There was lacking of combing on Yanjing medical school and inheritance system. The inheritance of Yanjing medical school was done far from good, and the study on how to inherit Yanjing medical school need strengthening.

Objective To study the alternations of regulatory T cells in early HIV infected patients and its association with disease progression.Methods Fifty-one untreated HIV infected patients were enrolled and divided into 3 groups according to their infection time and CD+4 T cell levels(30 early HIV infected patients,15 typical progressors,6 AIDS patients).Twenty normal controls were enrolled.There were no significant differences between the age and sex among four groups.Blood was drawn by venipuncture from each subject in EDTA tubes and the levels of CD+4 CD+25 Foxp3 + regulatory T cells were detected by FACSAria flow-cytometry.Spearman correlation was used to detect association between CD+4 CD+25 Foxp3 + regulatory T cells and the absolute CD+4 T cells,viral load and activation of T cells.Results The levels of CD+4 CD+25Foxp3+ regulatory T cells showed the tendency of increasing tendency from normal control to early HIV infected patients,asymptomatic HIV infected patients and AIDS patients.Early HIV infected patients was significantly lower than that in AIDS group [3.79(2.11 - 5.43) % vs 8.09(4.90 - 8.90) %,Z = - 2.29,P = 0.022].The levels of CD+4 CD+25 Foxp3 + Treg cells were associated with viral set point(r = 0.479,P =0.038) and inversely associated with CD+4 T cells(r = -0.455,P =0.011) and closely associated with HLA-DR expression on CD+3 T cells(r = 0.533,P = 0.002).Conclusions The ratio of CD+4 CD+25 Foxp3 +regulatory T cells of early HIV infected patients was significantly increased and associated with viral set point and CD+4 T cell counts,which indicate that alternation of regulatory T cell may be an important factor contributing to the disease progression in early HIV infection.

Objective To investigate the level of B7-H1 and its counter-receptor PD-1 expression in mDC and different subsets of T lymphocytes in HIV infected individuals in China and to analyze the correlation between the level of B7-H1/PD-1 and disease progression, and to demonstrate that PD-1/PD-L1-dependent inhibition is operating in HIV infected patients.Methods Percentage of B7-H1 and PD-1 expression in mDC, CD+4 T cells and CD+8 T cells from thirty-six untreated HIV infected patients and 20 health controls were selected and detected by flow-cytometry, its correlations with CD+4 T cell absolute counts and plasma viral loads were analyzed.Results The percentage of B7-H1 expression in mDC, CD+4 T cells and CD+8 T cells (mean 15.21, mean 20.63, mean 13.5)were higher than that of health controls (all P<0.05).The percentage of PD-1 expression in CD+4 T cells and CD+8 T cells (mean 17.91, mean 19.21)were higher than that of health controls (P<0.05, P<0.05). The level of B7-H1 and PD-1 expression were inversely correlated with CD+4 T-cell counts(mDC+B7-H1+:r=-0.647, P<0.01;CD+4B7-H1+:r=-0.489, P=0.002;CD+8B7-H1+:r=-0.372, P=0.026;CD+4PD-1+:r=-0.374, P=0.025;CD+8PD-1+:r=-0.455, P=0.005) and positively correlated with HIV viral load(mDC+B7-H1+:r=0.662, P<0.01;CD+4B7-H1+: r=0.426, P=0.01;CD+8B7-H1+:r=0.531, P=0.001;CD+4PD-1+:r=0.362, P=0.03;CD+8PD-1+:r=0.380, P=0.022).Conclusion The level of B7-H1 and PD-1 expression was associated with HIV disease progression, which provides a useful marker to define disease progression of HIV infection.

Objective To explore neurocognitive characteristics of human immunodeficiency virus (HIV)-infected patients ,and to compare the efficacy of highly active antiretroviral therapy (HAART ) among patients with different cognitive functions .Methods Cognitive function was evaluated using the Montreal cognitive assessment (MoCA) Chinese version in 118 HIV-positive patients and 62 HIV-negative controls .Among 59 patients on HAART ,CD4 + T cell count and viral load were assessed at enrollment and one-year follow-up .The mean of measurement data was compared using t test ,and enumeration data was analyzed using chi-squared or Fisher exact test when appropriate .Univariate and multivariate analysis were examined using bivariate Logistic regression models .Results Compared with control group ,HIV-infected group was characterized by higher rate of neurocognitive impairment (46 .6% vs 12 .9% , t =20 .30 ,P< 0 .05) ,and generally lower MoCA subscores for visuospatial abilities ,the clock drawing test , naming ,attention ,abstraction and delayed recall (t= - 3 .761 , - 2 .638 , - 4 .263 , - 3 .769 , - 3 .858 and- 3 .111 ,respectively ,all P< 0 .05) .Among patients on HAART ,subjects who scored < 26 showed no significant differences in viral load at three time points (pre-HAART ,post-HAART at enrollment and one-year follow-up) with those who scored ≥ 26 (t = 0 .557 ,0 .737 and - 0 .758 ,respectively ,all P >0 .05) .The former group had lower CD4 + T cell counts both at enrollment ([286 ± 127]/μL vs [363 ± 160]/μL) and one-year follow-up ([334 ± 122]/μL vs [411 ± 152]/μL) than the latter group (t= - 2 .027 and - 2 .067 ,respectively ,both P < 0 .05 ) ,while there were no obvious differences of pretreatment CD4 + T cell counts ([135 ± 77]/μL vs [155 ± 80]/μL) and HAART duration ([22 .29 ± 21 .20] months vs [18 .74 ± 16 .63] months) between these two groups (t= - 0 .968 and 0 .702 ,respectively ,both P>0 .05) .Multivariate analysis revealed that age (OR = 1 .044 ,95% CI :1 .008 - 1 .081 , P < 0 .05) and education time (OR = 0 .820 ,95% CI :0 .723 - 0 .930 , P < 0 .05 ) were independent predictors for neurocognitive impairment among HIV-infected patients . Conclusions Neurocognitive impairment is common among HIV-infected patients ,which is characterized by poor performance in multiple domains , and patients with neurocognitive impairment performed poorly in immune recovery .MoCA could be a useful screening tool of cognitive function in HIV-infected patients . Neurocognitive function has no relationship with pre- and post-treatment viral levels .

Objective:To study the changes of NKG2C/NKG2A expressed on T cells in HIV chronically infected individuals and HAART-treatment AIDS patients and the relationship with disease progression of HIV. Methods: We collected peripheral blood from HIV chronically infected individuals,HAART-treatment AIDS patients and healthy human and used the flow cytometry by staining fluorescent antibody to detect the NKG2C/NKG2A receptors expressed on T cells. Results:NKG2C+,NKG2A+ and NKG2C+NKG2A-expressed on T cells in HIV chronically infected individuals were significantly higher than the healthy control group ( P=0. 025,P=0. 032,P=0. 029),while in HAART-treatment AIDS patients were significantly lower than that in HIV chronically infected individuals (P=0. 033,P=0. 037,P=0. 018),returned to the normal levels with no significant difference compared with the healthy control group. The absolute number of peripheral blood CD4+ T lymphocytes in HIV chronically infected individuals was negative correlation with T cells which expressing NKG2A+,NKG2C+NKG2A+ and NKG2C-NKG2A+( r=-0. 697,P<0. 000 1;r=-0. 463,P=0. 015;r=-0. 693,P<0. 000 1). What was more,the absolute number of peripheral blood CD4+ T lymphocytes had positive correlation with the ratio of NKG2C and NKG2A expressed on T cells receptor in HIV chronically infected individuals(r=0. 476,P=0. 012). Conclusion:Studying the expression of NKG2C and NKG2A receptors on T cell has great significance in HIV infected individuals, which may provide a scientific basis for clinical prognosis of HIV infection.

Objective To investigate the killer cell lg-like receptors (KIR) gene frequency of HIV-1 infected slow progressors(SP) and typical progressors(TP), and to analyze the interaction between KIR alleles and the progression of HIV-1 infection in Chinese population. Methods Eighty-one HIV-1 posi-tive individuals including 43 SPs and 38 TPs were recruited. Carriage of KIR genes was assessed using poly-merase chain reaction sequence-specific primers (PCR-SSP) assays. Results KIR2DS3 gene frequency was significantly lower in SP group (3.6%) than that in TP group (14.2%), P =0. 018 ,OR =0. 210,95% CI =0.053-0.833. The number of activating KIR genes was less in SP group than that in TP group, but was not significant (P = 0. 208). Conclusion Lower KIR2DS3 gene frequency may potentially be associated with slower progression to AIDS in Chinese population.