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.

ObjectiveTo investigate the expression of stem cell marker adenosine triphosphate (ATP)-binding cassette transporter 2 (ABCG2) in the tissue and side population (SP) of a cell line A431 of human cutaneous squamous cell carcinoma(SCC).MethodsSP cells were separated by flow cytometry from cultured A431 cells.Methyl thiazolyl tetrazolium(MTT) assay was used to evaluate the proliferative ability of,reverse transcription-PCR to determine the expression of ABCG2 in,SP and non-SP cells.Immunohistochemistry (MaxVision method) was carried out to detect the expression of ABCG2 protein in tissue specimens from 10 patients with SCC.ResultsSP cells existed in cultured A431 cells,and accounted for about 1.1％ of A431 cells.The SP cells had a stronger growth and colony-forming ability than non-SP cells did.The number of cell clones formed by SP cells and non-SP cells was 114.8 ± 4.95 and 44.5 ± 3.67,respectively,per well in a 24-well plate( t =27.92,P ＜ 0.01 ).The expression level of ABCG2 mRNA was significantly higher in SP cells than in non-SP cells(t =5.22,P＜ 0.01).There existed a small number of ABCG2 positive cells in SCC tissue,and ABCG2 was mainly expressed in the cytoplasm and membrane of tumor cells.ConclusionsSP cells exist in A431 cells,which have characteristics of stem cells and highly express ABCG2.ABCG2 may be a potential stem cell surface marker of SCC.

Objective To explore the clinical efficacy of image-guided 125I radioactive seed interstitial implantation therapy for unresectable pancreatic cancer.Methods 25 patients with unresectable pancreatic cancer evaluated by retrospective follow-up were enrolled in this study,13 patients received radioactive seeds implantation while 12 patients were given non-surgical treatment.We observe and compare the clinical benefits,objective curative effect,complications,adverse reaction,survival between the two groups of patients.Results Compared with the non-surgical treatment group,the clinical benefit rate in the radiotherapy seed implantation group was 92％ (12/13) while that of the non-surgical group was 42％ (5/12),the difference was of statistically significance.The numbers of cases evaluated as effective were 6 (46％) and 4 (33 ％) respectively,the difference was not statistical significant (x2 =0.427,P ＞ 0.05);The radioactive seed implantation group had no serious postoperative complications;3 cases who received subsequent chemotherapy in radioactive seed implantation group(23％,3/13) and 3 cases in non-surgical treatment group(25％,3/12)suffered from serious adverse reactions,the difference was of no statistical significance(x2 =0.013,P ＞0.05);Comparing the survival rate between the two groups,x2 =0.001,P =0.969,the difference was of no statistical significance.Conclusions The therapy of 125I radioactive seed implantation for unresectable pancreatic cancer significantly relieves cancer caused pain and improves quality of life.

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 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 .

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 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 .