The study is to describe the main activities of NGOs involved in programs of China AIDS Global Fund Round 5 and analyze the advantage and necessity of their involvement in HIV/AIDS control. The experience and problems of NGOs ' involvement in AIDS Global Fund Programs,as well as appropriate approaches to address them are discussed.

The effects of microRNA-34a (miR-34a)-regulated Notch1 gene on the proliferation and apoptosis of the human glioma cell line U87 were investigated in this study. The U87 cells were divided into miR-34a mimics, negative control, mock transfection and blank control groups in terms of different treatments. In miR-34a mimics group, human U87 glioma cells were transfected with miR-34a mimics by using lipofectamine 2000. The cells transfected with nonsense microRNA were set up as negative control group. Those treated with lipofectamine 2000 only were designated to the mock tranfection group. In the blank control group, the cells were cultured routinely and no treatment was given. The expression of miR-34a and Notch1 was detected by using real-time RT-PCR. Western blotting was employed to monitor the change in Notch1 protein. Cell proliferation and apoptosis were measured by CCK-8 and flow cytometry. The results showed that the proliferative ability of U87 cells was significantly reduced and the apoptotic cells increased in miR-34a mimics group relative to control groups. The expression of miR-34a was significantly up-regulated in mimics group as compared with control groups (P<0.05). Furthermore, Notch1 protein levels were significantly decreased in miR-34a mimics group when compared with control groups (P<0.05), but the mRNA expression of Notch1 showed no significant difference among these groups. It was concluded that miR-34a may suppress the proliferation and induce apoptosis of U87 cells by decreasing the expression of target gene Notch1, suggesting that miR-34a may become a promising gene therapeutic target for brain glioma.

Objective To explore the influencing factors for the purity and viability of primary cultured cortical neurons of rat,and optimize the separating and cultivating conditions of the cortical neurons.Methods The primary cotical neurons were cultured in a serum-free culture system of B27-supplemented neurobasal medium.The differences in purity and viability of primary cultured neurons between embr-yonic rat group and newborn rat [postnatal 24 h and 5 d] group were evaluated by morphology,immunocytochemistry of neuron-specific enolase(NSE) and trypan blue staining.The changes of neurons purity and viability in different trypsin digestion time(0 min,5 min and 15 min) at different environment temperatures(20 ℃ and 30 ℃) were assessed by immunocytochemistry and trypan blue staining.Results The primary cultured neurons from fetal and newborn rats grew well.There was no significant difference in embryonic rat and postnatal 1 d newborn rat group[(91.30?1.03)%,(89.50?1.78)% respectively in purity;and(98.20?0.58)%,(97.10?0.98)% respectively in viability].The neurons from 5-days newborn rat were inferior to that from fetal and 1-day newborn rat in purity and viability[(82.00?1.25)% and(92.87?1.56)% respectively].Shortening operation time and adjusting digestion time according to the environment temperatures could improve neuronal viability:A digestion for 15min at the environment temperature of 20 ℃ or for 5 min at 30 ℃ could acquired cells with higher viability [(98.20?0.58)% and(96.70?0.64)% respectively].Conclusions It is an easy,practical choice to culture priamry cortical neurons from postnatal 1 d newborn rats.Optimizing the separating and cultivating condition(environment temperatures,digest time,et al.) will improve the neurons purity and viability.

AIM:To determine the contents of isofraxidin and rosmarinic acid in Zhongjiefeng Tablet(Sarcandra glabra(Thunb)). METHODS:HPLC was performed on Diamonsil C18 column(250 mm ? 4. 6 mm,5 ?m),the mobile phase consisted of acetonitrile-0. 1% phosphoric acid solution(80 ∶ 20) at a flow rate of 1. 0 mL/min. The column temperature was set at 35 ℃,UV detection wavelength was at 342 nm. RESULTS:Contents of isofraxidin and rosmarinic acid detecded showed good linear relation(R2 =0. 999 99,0. 999 92,respectively) and the average recoveries(n =6) of 98. 7% (RSD =2. 22% )and 96. 9% (RSD =2. 76% ). CONCLUSION:The method is fea-sible,accurate and reliable,thereby available for quality control.

Insulin receptor substrate(IRS)-1 and IRS-2 expression levels of liver tissues and skeletal muscle in intrauterine growth retarded(IUGR)rats were investigated by RT-PCR and immunohistochemistry.An IUGR animal model was established by maternal nutrition restriction during pregnancy.IRS-2 expression level of liver tissue and IRS-1 expression level of skeletal muscle in IUGR rats at 0 and 3 weeks old were significantly lower than those in normal rats at the same age respectively,and insulin resistance was induced in IUGR,and these findings might be the molecular mechanisms susceptible to metabolic syndrome in IUGR rats.

Objectives To investigate dynamic pathological features and virus distribution in the liver with a murine severe acute respiratory syndrome(SARS)model injected with murine hepati- tis virus 3(MHV-3)through trachea.As a representative of host genes,mouse fgl2(mfgl2)pro- thrombinase gene expression and its clinical significance were discussed in SARS associated liver dam- ages.Methods The Balb/cJ mice were infected with 100 PFU of MHV-3 through trachea and Balb/ cJ mice injected with saline were served as control.Survival rate,pathological features in organs and liver function were observed.Virus titers in different organs were determined on monolayer of L2 cells by a standard plaque assay.Virus distribution and cellular localization were studied by in situ hy- bridization.Both mfgl2 and fibrin expressions were examined in the liver by in situ hybridization and immunohistochemistry to investigate the role of mfgl2 in the liver impairment.Results Mice infected with MHV-3 through trachea developed multiple organs damages and died within 5 days,while all mice in control group survived with no histopathological changes.Infected liver tissues showed wide- spread cloudy swelling,prominent ballooning degeneration with mild lymphocytic infiltration in the portal area.Dot and zonal hepatocellular necrosis could be found occasionally.The lungs showed typi- cal interstitial pneumonia and hyaline membranes formation.Other histological changes also could be found in other organs examined.MHV-3 virus replication was identified in all organs observed.The liver function was injured,mfgl2 expression were evidenced mainly in the necrosis areas with fibrin deposition around the necrosis areas.Conclusions Pathological changes of the liver in this murine SARS model can mimic the liver impairment characteristics of SARS in human.In addition to the physical damage induced by the virus,the up-regulation of novel gene mfgl2 in the liver in association with fibrin deposition may play a vital role in the development of SARS associated liver damages.

To study the contribution of T cell subsets in the pathogenesis of Murine hepatitis virus Type3 (MHV-3) induced chronic viral hepatitis in C3H/Hej mice, ninety C3H/Hej mice were chosen to individually receive 10 plaque forming units (PFU) of MHV-3 intraperitoneally. The changes of virus titer and pathology in liver tissue were examined by standard plaque assay and by the hematoxylin/eosin (HE) staining method from 2 days post MHV-3 infection. The ratios of T cell subsets including CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4-CD8-, CD3+CD4+CD25+, CD3+CD4+CD25- and CD3+CD4-CD25+ T lymphocyte of total T lymphocytes in blood, spleen and liver were examined at 0, 2, 4, 6,8, 10, 12, 15, 20, 25, 30, 40 days post MHV-3 infection by flow cytosorting. We observed that the virus titer raised and showed persistent virus duplications and inflammatory changes in the livers of C3H/Hej mice from 2 days post MHV-3 infection. The double negative T cell (DN Treg cell) and CD4+CD25+ T cell ratios increased significantly from 2 days post MHV-3 infection in C3H/Hej mice, and CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4+CD25- and CD3+CD4-CD25+ T cell ratios decreased accordingly. In conclusion, the changes of virus titer and pathology in the livers of C3H/Hej mice post MHV-3 suggest their contribution to viral persistence. Further characterizations of DN Treg cells are that infection indicates that MHV-3 could induce the chronic inflammation in livers of C3H/Hej mice.The increase of the DN Treg cell and CD4+CD25+ T cell ratios in C3H/Hej mice post MHV-3 infection suggests that DN Treg cells and CD4+CD25+ T cells may both have important suppressive immunomodulation functions in the development of chronic viral hepatitis and have important roles in the virus persistent infection. Further characterizations of DNT cell and CD4+CD25+ T cell are under investigation.

Objective To study the effect of nickel-titanium stent(NTS) and consequent anti-allergy dexamethasone therapy on macrophage cells reactivity to lipopolysaccharide (LPS) from gram-negative bacterium .Methods The macrophage cell line Raw 264 .7 and dexamethasone-pretreated Raw264 .7 were co-cultured with NTS for 4 days ,and stimulated with LPS for 24 hours .The surface marker CD molecules of CD80 ,CD86 and FasL were detected with flowcytometr method ,the supernant cytokine production of proinflammatory cytokines IL-6 and TNF-αwas valued with ELISA method ,and intracellular inflammatory signal pathway acti-vation of NF-κB ,GAS ,ISRE and STAT3 was checked with signal molecule specific promoter lunciferase analysis .Results The stent pre-treatment improved LPS-mediated CD80 expression ,suppressed FasL production ,decreased IL-6 secretion and NF-κB ac-tivation ,the results have statistical significance (P<0 .05) .The dexamethasone treatment improved stent-mediated up-regulated ex-pression of CD80 ,FasL and TNF-α,and suppressed the activation of intracellular inflammatory signal pathway of NF-κB ,ISRE and STAT3 ,the results have statistical significance(P<0 .05) .Conclusion NTS inhibit macrophage cells Raw264 .7 react to TLR4 ag-onist LPS ,and dexamethasone treatment improved the function .

To evaluate the role of murine fibrinogen like protein 2 (mfgl2) /fibroleukin in lung impairment in Severe acute respiratory syndrome (SARS), a murine SARS model induced by Murine hepatitis virus strain 3 (MHV-3) through trachea was established. Impressively, all the animals developed interstitial pneumonia with extensive hyaline membranes formation within alveoli, and presence of micro-vascular thrombosis in the pulmonary vessels. MHV-3 nucleocapsid gene transcripts were identified in multiple organs including lungs, spleen etc. As a representative proinflammatory gene, mfgl2 prothrombinase expression was evident in terminal and respiratory bronchioles, alveolar epithelia and infiltrated cells in the lungs associated with fibrin deposition and micro-vascular thrombosis. In summary, the established murine SARS model could mimic the pathologic characteristics of lungs in patients with SARS. Besides the physical damages due to virus replication in organs, the up-regulation of novel gene mfgl2 in lungs may play a vital role in the development of SARS associated lung damage.

Objective To compare the clinical utility of alpha-fetoprotein (AFP) and des-gammacarboxyprothrombin (DCP) in diagnosing hepatocellular carcinoma (HCC) in patients with a hepatic mass.Methods From January 2015 to May 2015,141 patients were diagnosed to have a liver tumor after imaging examinations in the Hepatobiliary Surgical General Hospital of PLA,Beijing,China.Preoperative AFP and DCP were measured using commercial assay kits.The reference standard was either pathologic or clinical diagnosis of HCC.The performance of AFP and DCP in diagnosing HCC was determined using receiver operating characteristic curve analysis.Results Of 141 patients,98 were diagnosed to have HCC and 43 without.The levels of AFP were significantly higher in patients with HCC than those without [80.0(3.9-1 375.0) μg/L vs.2.1 (1.6-3.2) μg/L,Z =6.98,P ＜ 0.01].Similar results were observed in the levels of DCP [141.5 (24.0-978.0) AU/L vs.19.0 (14.0-25.5) AU/L,Z =5.18,P ＜ 0.01].Receiver operating curves (ROC) indicated the cut-off value with the best sensitivity and specificity was 3.6 μg/L for AFP and 35 AU/L for DCP.The difference in the area under ROC between AFP and DCP was not statistically significant (0.87 vs.0.78,Z =1.72,P =0.085).The sensitivity and specificity for detection of HCC in patients with a hepatic mass were 56.1％ and 95.4％ for AFP ＞ or =20 μg/L,69.4％ and 83.7％ for DCP ＞ or =40 AU/L,respectively.The level of AFP was associated with DCP in patients with HCC (x2 =9.12,P ＜ 0.01,r =0.292) and parallel testing of AFP and DCP gave an optimal sensitivity of 79.6％ with a specificity of 81.4％ in diagnosing HCC.Conclusions DCP is a useful biomarker and it gave an equal performance as AFP in diagnosing HCC in patients with a liver mass in this study.Parallel testing of AFP and DCP effectively increased the diagnostic sensitivity.Although the biomarkers only marginally improved the diagnostic results,it could be useful in diagnosing HCC in individuals who had atypical imaging results.