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.

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.

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.

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.

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.

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 correlation between drug-resistance of paclitaxel and the expres- sion levels of anti-apoptotic protein survivin and the role of matrine reversal resistance of PTX in non-small cell lung cancer.Methods The expressions of survivin in 120 samples of human lung cancer with paclitaxel chemotherapy were detected by immunohistochemical staining.Results The positive expression rate of sur- vivin was 57.5%.In the positive expression of survivin group,chemotherapy combined matrine could improve response rate and survival time(P0.05).Conclusion The survivin expression was related to the response rate of paclitaxel in lung cancer.It might be a valuable new marker to predict the drug-resistance of paclitaxel.In this prospective research,survivin protein,which promoted the apoptosis caused by paclitaxel,may be the target of martine.It could partly reverse the drug re- sistance to paclitaxel.

AIM:To explore a more accurate and reliable pathological model of the chronic bronchitis , which has improved from the former single-factor modeling method of the disease .METHODS:The mice in complex group were treated with lipopolysaccharide ( LPS) by tracheal injection on the 1st day and nasal drops on the 14th day, and from the 2nd day to 30th day, the animals were given passive smoking and sulfur dioxide ( SO2 ) inhalation ( except on the 14th day).The mice in SO2 group were exposed to SO2 2 min per day, while in smoking group, the mice were exposed to smoke for about 1 h per day (4 cigarettes each time until one pack of cigarettes were burning up ).In LPS group, the mice had tracheal injection of LPS on the 1st day and nasal drops of LPS on the 14th day and 30th day.Every modeling process las-ted for 30 days.After modeling, the improvement of chronic bronchitis model was evaluated by testing the general condi-tions of the mice , analyzing leukocyte count in bronchoalveolar lavage fluid ( BALF ) , and observing the morphological changes of the bronchial and lung tissues .RESULTS:After modeling, the mice in every model group experienced symp-toms including wet nose, cough, dry and lusterless hair, arched back and curled-up body, showing inactive, and slow down in response .The mice in complex group gained the lowest weight compared to other groups .From each model group , the inflammatory cells infiltrated evidently around the bronchial walls , especially in the bronchial cavity , and the mucilage secretion in the airway increased .The total number of leukocytes in BALF increased significantly in complex group .The in-flammatory cell count in the lung tissue indicated that the mice in complex group had significantly higher levels of inflamma -tory cell infiltration.Besides, the comparison between smoke group and LPS group was statistically significant .CONCLU-SION:Smoking, SO2 inhalation and LPS injection induce bronchial lung disease in mice , and the complex chronic bron-chitis mouse model is a better model with the pathological changes of bronchus , lung tissue and BALF , and pathogenesis of chronic bronchitis .

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.