Objective:To investigate the expression and significance of ABCG2(the G2 member of ATP-binding cassette family)in U251and its stem cell.Methods:The U251 were cultured in serum-free medium with EGF,bFGF,LIF and B27.The tumor spheres formed and cultured continuously in the serum-free medium through the monoclonal formation experiment.The immunofluorescence staining was employed to identify the U251 stem cell.The expression of ABCG2 was detected by confocal microscopy and Western blot.Results:The stem cells from malignant glioma cell line U251 have been successfully isolated.The cells expressed CD 133.The ABCG2 protein positive stain was located in the cell membrane in both U251 cells and its stem cell with confocal microscopy method.Western blotting showABCG2 was expressed in U251 cells and in its stem cells.The expression level in the stem cells was significantly higher than U251 cells.Conclusion:ABCG2 is expressed in U251 cells and in U251 stem cells.The expression level in the latter was higher.

A total of 3 177 HIV/AIDS patients were admitted in Beijing Ditan Hospital,Capital Medical University from January 2009 to December 2015,among whom pneumothorax developed in 50 cases with a morbidity rate of 1.6％.Twenty six HIV/AIDS patients with pneumothorax died with a case fatality rate of 52.0％ (26/50).Pneunocystis jirovecii pneumonia (PCP) was the dominant lung disease related to pneumothorax (37/50).Risk factors of pneumothorax were assessed among 40 HIV/AIDS patients with PCP undergoing mechanical ventilation in ICU,including 20 cases with pneumothorax and 20 cases without pneumothorac.Multivariate logistic regression analysis revealed that positive end-expiratory pressure (PEEP) was independent risk factor of pneumothorax in HIV/AIDS patients with PCP under mechanical ventilation (OR =2.490,95 ％ CI:1.302-4.763,P =0.01).

AIM:To study the mechanisms of effects of the ?-opioid receptor(?-OR) on neonatal rat hypertrophic myocardial cells induced by isoproterenol(ISO).METHODS:Myocardial cells of neonatal rats were cultured in vitro.The total protein content was detected by the Lowry's assay.The volume of myocardial cell was measured by computer photogragh analysis system and the protein synthesis was assayed with[3H] leucine intake method.The degree of extracellular signal-regulated kinase(ERK) phosphorylation was determined by western-blotting.RESULTS:U0126(an inhibitor of ERK) decreased the total protein content,the cell size,the[3H]leucine incorporation and the ERK phosphorylation degree in hypertrophic myocardial cells induced by ISO.1 ?mol/L U50,488H(a ?-opioid receptor antagonist) could decrease the total protein content,the cell size,the[3H]leucine incorporation and the degree of ERK phosphorylation.But these effects could be diminished when incubated with U0126.CONCLUSION:?-opioid can abolish the hypertrophic response induced by ISO of cardiac myocytes,which was associate with ERK pathway.The inhibitor of ERK significantly inhibits the ISO-induced hypertrophied effect.

Objective To investigate the proper inner control genes suitable for mRNA expression level comparison of aging rat tissues.Methods Real-time reverse transcription PCR was used to examine in aging rat tissues the expression level of G3pd(glyceraldehyde-3-phosphate dehydrogenase),ACTB(?-actin),H3f3b(H3 histone,family 3B),Arbp(acidic ribosomal phosphoprotein P0)and 18S(18S ribosomal RNA).Results The most stably expressed housekeeping gene in aging rat kidney was ACTB,in heart and lung G3pd showed the minimum variation;Arbp expression was the most stable one in different tissues.Conclusion For aging rat intra-tissue mRNA normalization at least two housekeeping genes should be used: one is the ribosomal RNA gene 18S and another one is Arbp.

Objective To study full-automatic tissue hydroextractor that its factors of influence anhydration quality in the process of anhydration. Methods The quality of tissue hydroextractor was increased by various angles of reagent examination, correct clipping and standard management etc. Results The quality of hydroextrator and the high quality of tissue section was improved by replacing reagents timely, clipping tissue properly, canonical management and performing specification and maintenance of the machinery, and the efficiency and accuracy of pathological diagnosis was enhanced too. Conclusion In foundation of strictly quality control, the advanced machine integrates with the skill and sense of responsibility of the operators. The quality of tissue dehydration can be improved effectively.

Objective To investigate the effects of ATP1A1 knockdown by RNA interference(RNAi) on proliferation of human U251 glioma stem cells .Methods The human U251 glioma stem cells were infected with lentivirus expressing ATP1A1-shRNA . The mRNA and protein expressions of ATP1A1 in U251 glioma stem cells were detected by RT-qPCR and Western blotting ,re-spectively .The cell cycle and apoptosis were evaluated by flow cytometry .The proliferation of U251 glioma stem cells was deter-mined by MTT assay .Results The expressions of ATP1A1 in U251 glioma stem cells transfected with ATP1A1-shRNA were in-hibited significantly at both mRNA and protein levels ,with an inhibitory rate of 84 .15% for ATP1A1 mRNA and of 55 .33% for ATP1A1 protein respectively .The proliferation of cells was inhibited ,the cell apoptotic rate was significantly increased and the cell cycle was arrested in G1 phase and S phase decreased significantly in ATP1A1-shRNA cells(P<0 .05) .Conclusion RNAi targe-ting ATP1A1 gene could down-regulates the ATP1A1 expression ,induces cell apoptosis ,regulates cell phase redistribution and in-hibits cell proliferation in U 251 glioma stem cells .

Objective To evaluate the effect of HIV infection and highly active antiretroviral therapy (HAART) on mitochondrial function and mass in peripheral monocytes.Methods There were 14 ART-naive HIV-infected patients,14 NRTI treated HIV-infected patients and 12 healthy controls from Beijing Ditan Hospital.The mitochondrial membrane potential and mitochondrial mass in monocytes were analyzed by flow cytometry.Mitochondrial disturbances related to HIV infection and HAART in monocytes were analyzed.Results In ART-naive patients and NRTI-exposed patients,the levels of mitochondrial membrane potential in monocytes (77.74 ± 14.77,73.94 ± 12.87) were significantly lower than these in healthy controls (89.43 ±4.06) (P =0.032 8,P =0.002 6).The amount of mitochondrial mass in NRTI-exposed patients (3 329.0 ± 836.7) was significantly higher than that in healthy controls (2 075.0 ± 932.2) and that in ART-naive patients (2 592.0 ± 781.5) (P ＜ 0.05).Conclusions The abnormal of mitochondrial membrane potential and mitochondrial mass in monocytes from HIV-infected patients were related to HIV infection and the introduction of HAART.

Objective:To study the clinical features and treatment outcome of AIDS associated oral candidiasis.Methods:The clinical data of 31 cases with AIDSassociated oral candidiasis from 201209 to 201303 were studied retrospectively,including general data,clinical features,oral manifestation,CD4 cell count,opportunistic infections,and antifungal therapy outcome,etc.Results:CD4cell count ＜200 cell/μl was found in 30 cases,AIDSrelated multiple opportunistic infection was observed in 29 cases.30 cases hadpseudomembranous candidiasis,1 cases had erythematous candidiasis and 2 cases had pseudomembranous candidiasis with angular candidiasis.After antifungal treatment,the lesion of 8 cases reduced,that of 23 cases disappeared completely,lesion relapse after drugwithdrawal happened in 3 cases.Conclusion:AIDSassociated oral candidiasis was more common in AIDS patients with CD4 ＜200cells/μl,the main clinical form is pseudomembranous type,and with multiple opportunistic infections.The antifungal treatment is effective for the patients.

Objective To identify the relationship between impaired glucose regulation of neuronal apoptosis in hip-pocampus and the ability of learning/memory in rats.Methods The model rats were made by high fat and sugar diet;The morris water maze was used to detect the learning and memory function;The TUNEL method was used to detect neuronal apoptosis in hippocampus .Expression of Bcl-2/Bax and Bcl-2 mRNA/Bax mRNA factor in hippo-campus neurons was detected with immunohistochemistry and in situ hybridization .Results Compared with NGT group, in IGR group the learning and memory ability were meaningfully decreased (P<0.05);the number of neuro-nal apoptosis in hippocampus was increased significantly ( P<0.05 ); the expression of Bcl-2 and Bcl-2 mRNA in hippocampus decreased significantly ( P<0.05);the expression of Bax and Bax mRNA in hippocampus increased sig-nificantly ( P<0.05 );The ability of learning and memory was positively correlated with expression of Bcl -2 ( P<0.05) and negatively correlated to the expression of Bax (P<0.05).Conclusions There is a relationship between impaired glucose regulation and the ability of learning and memory in rats , its mechanism may be potentially related to hippocampal neuronal apoptosis .