Objective To investigate the effects of inhibition of STAT5 gene expression by RNA interference technology on apoptosis of human hepatocellular carcinoma cell line SMMC-7721. Methods Three siRNA eukaryotic expression vectors against STAT5 were constructed and transfected with lipofectamineTM 2000 into SMMC-7721 cells. The changes in STAT5 expression were detected by semi-quantitative RT-PCR and Western blot. Cell apoptosis was assayed by flow cytometry (FCM). Results The sequence-specific siRNA could effectively and specifically inhibit STAT5 gene expression at both mRNA and protein levels. The inhibition rates of STAT5 mRNA expression were 70.43%, 43.02%, and 45.07%, respectively. The inhibition rates of STAT5 protein expression were 67.45%, 37.36%, and 41.86%, respectively. At 48 h after transfection, apoptosis rate was 25.61%. Conclusion siRNA against STAT5 can inhibit STAT5 gene expression in SMMC-7721 cells effectively and specifically and induce apoptosis of SMMC-7721 cells. siRNA targeting STAT5 has a great potential value in gene therapy of hepatocellular carcinoma.

Objective To examine clusters of anti-nuclear antibodies (ANA) and their associations with clinical features in patients with systemic lupus erythematosus (SLE).Methods It was a retrospective study.113 SLE patients were reviewed from March 2010 to May 2012 in Department of Rheumatology,Jinhua Central Hospital.ANA and specific autoantibodies to 15 kinds of nuclear antigens were tested by indirect immunofluorescence assay (IIF) and line immunoassay (LIA) respectively.Hierarchical clustering method was performed to analyze specific clusters of ANA profiles in SLE.Chi-square tests were used to investigate relationship between antibody clusters and clinical features of SLE.Results The positive rate of LIA for ANA was 97.3％,consistent with IIF method,and the total accordance rate of the both methods was 98.2％.Thirteen kinds of antigen-specific antibodies were detected in SLE patients by LIA.Clustering analysis for these antibodies showed three specific clusters in SLE,Nuc/His/dsDNA cluster (C1),low-Ro/low-La cluster (C2),and Ro/Sm/RNP cluster (C3),accounting for 36.3％,24.8％,38.9％ of the total cases respectively.There were significant difference of AST levels among three clusters [(32.62 ± 21.92)U/L,(25.56 ± 16.63) U/L,(50.41 ± 60.86) U/L respectively for C1,C2 and C3].High incidences of chronic cutaneous lupus,abnormal renal indicators and inflammatory synovitis were found in all three clusters.Besides,there were significant differences among three clusters for the incidences of chronic cutaneous lupus (39.0％,39.3％,63.6％ respectively for C1,C2,C3) and leukopenia/lymphopenia (56.1％,25.0％,56.8％ respectively for C1,C2,C3) (P ＜ 0.05).Patients in Ro/Sm/RNP cluster showed higher incidences of lupus nephritis (43.2％/26.8％ or 39.3％); patients in low-Ro/low-La cluster showed low risk of hypertension (7.1％/19.5％ or 22.7％) ; patients in Nuc/His/dsDNA cluster showed high incidences of thrombocytopenia (41.5％/21.4％ or 25.0％) and high risk of lung or upper respiratory tract infection (46.3％/28.6％ or 29.5％),but low incidence of neurologic symptoms (0％/ 3.6％ ％ or 11.4％).Conclusion Three characterized ANA clusters are identified in SLE patients in this pilot study.Different clusters are associated with certain clinical features and complications ofSLE.However,the correlations found in this study need to be investigated further in larger populations.

0.05],significantly higher than stage Ⅲ,Ⅳ[60%(6/10),50%(5/10);P

Acute Disease ; Adolescent ; Female ; Heart Neoplasms ; complications ; Humans ; Hypertrophy, Left Ventricular ; etiology ; Myocardial Infarction ; etiology ; Myxoma ; complications

OBJECTIVETo explore the effect of internal fixation with screw through femoral epiphyseal plate on growth in- hibition via an experimental study.

METHODSForty New Zealand rabbits were randomly divided into 4 groups and 10 rabbits in each group. Epiphyseal plate was injured by penetrating of screws, and the size of damage area was controlled by changing the number of threads. Group A: blank group; group B: injury area accounted for 4% of the epiphyseal plate; group C: injury area accounted for 6%; group D: injury area accounted for 8%. The internal fixation was removed after 2 weeks, and the results were observed with X-ray film for 4 groups to judge the complications such as early closure of epiphyseal.

RESULTSIn each group, there were no statistical differences in the length of the femoral neck, the diameter of femoral neck, the diameter of the femoral head, and the epiphyseal plate closure time. The growth speed of the length and diameter of the femoral neck, as well as the diameter of femoral head, were quicker on the early phase, and the speed was slowest when the epiphyseal plate was being closed.

CONCLUSIONThe injury area of epiphyseal plate under 8% is safe for its growth. Because no evidences demonstrate the growth inhibition of epiphyseal plate, the screws can be used for rabbit epiphyseal plates.

Animals ; Bone Screws ; Female ; Femur Head ; growth & development ; surgery ; Fracture Fixation, Internal ; methods ; Growth Plate ; growth & development ; Magnetic Resonance Imaging ; Male ; Rabbits ; Salter-Harris Fractures

Objective To discuss the feasibility on building lewis lung carcinoma mouse models through different methods and improve the methods. Methods The method of culture LLC cells in vitro, trypsin digestion method, Ⅳ collagenase method and homogenate method were compared to make the different dose of cell suspension injected into C57BL/6 mice. The feasibility of the improved method was determined through observing the cell count, the tumor formation ratio, the tumor formation time, tumor volume, weight and life habit. Results The method of culture LLC cells in vitro could get needed cells and its tumor formation ratio was 100 %. Trypsin digestion method and homogenate method could get less cells and its tumor formation ratio was about 80 %～90 % and 60 %～75 %. Whereas 1V collagenase method could get most cell count and its tumor formation ratio was 100 %. Conclusion IV collagenase method is a preferred method which is simple,high efficiency and make a strong base on the cancer experimental study.

Adenoma, Pleomorphic ; diagnosis ; diagnostic imaging ; surgery ; Adolescent ; Adult ; Aged ; Angiography, Digital Subtraction ; Carcinoma, Adenoid Cystic ; diagnosis ; diagnostic imaging ; surgery ; Chemotherapy, Adjuvant ; Child ; Female ; Follow-Up Studies ; Head and Neck Neoplasms ; diagnosis ; diagnostic imaging ; surgery ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neurilemmoma ; diagnosis ; diagnostic imaging ; surgery ; Oral Surgical Procedures ; methods ; Postoperative Complications ; Radiotherapy, Adjuvant ; Tomography, X-Ray Computed ; Young Adult

BACKGROUND: Overexpression of phosphorylated Tau protein is a factor of dementia, and scholars abroad find that APP17 peptide may have effect on it.OBJECTIVE: To observe changes of phosphorylated Tau protein Ser202/Thr205 of mice with diabetes mellitus (DM) after injection of APP17 peptide.DESIGN: Randomized control study.SETTING: Department of Pathology, Capital University of Medical Sciences; Department of Brain Aging, Xuanwu Hospital, Capital University of Medical Sciences.MATERIALS: The experiment was carried out in the Pathological Department of Capital University of Medical Sciences and Brain Aging Department of Beijing Xuanwu Hospital. A total of 18 male Kunming mice of 8 weeks old and weighing 28-32 g were randomly divided into control group, DM group and APP17 peptide group with 6 in each group.METHODS: DM models were induced by streptozotocin (STZ) through selectively destroying β-islet cells; meanwhile, APP17 peptide was intraperitoneally injected into mice. Four weeks later, brain tissue underwentimmunohistochemical staining with AT-8 (Ser202/Thr205, a special monoclonal antibody).MAIN OUTCOME MEASURES: ① Morphological observation; ② AT-8 distribution; ③ quantitative analysis of immunohistochemical staining.RESULTS: Positive AT-8 cells in DM group were distributed in retrosplenial cortex, hippocampus, thalamus, hypothalamus, etc.; however, those incontrol and APP17 peptide groups were only distributed in retrosplenial cortex and hippocampus, and poorly stained.CONCLUSION: Positive AT-8 cells may be widely distributed in neurons of brains of DM mice; however, APP17 peptide may normalize the expression of positive AT-8 cells.

Objective To investigate the radiosensitization effects of the combination treatment of clioquinol (CQ) and zinc on human cervical cell line HeLa in vitro.Methods Cells were divided into the 4 groups:controls,drug,radiation,and combined drug and radiation group.Cytotoxic effect of CQ and zinc on cell viability was determined by CCK-8 assay.Radiosensitization effect of CQ and zinc on HeLa cells was detected by colongenic assay,and the single-hit multi-target model was used to stimulate the doseresponse curve of survival and to calculate radiosensitization parameters.The cell cycle and apoptosis of HeLa cells were analyzed with flow cytometry.Luciferase reporter assay was used to study NF-κB activity of HeLa cells.Results The combination of CQ and zinc inhibited cell growth in a dose-dependent manner (F =188.00,P ＜ 0.01).The mean lethal dose was 3.16 and 2.04 Gy for radiation group and combined drug and radiation group,respectively,and hence the SER was 1.55.Compared with the radiation group,the ratio of G2-phase cells in the combined drug and radiation group decreased(t =10.39,P ＜ 0.05),the apoptosis rate increased at 24 h post-irradiation (t =5.64,P ＜ 0.01),and the NF-κB activity decreased (t =21.42,P ＜ 0.05).Compared to the control group,the NF-κB activity increased in the radiation group(t=6.23,P＜0.05),but decreased in the drug group(t =12.48,P＜0.05).Conclusions The combination of CQ and zinc could increase the radiosensitivity of HeLa cells by decreasing the ratio of G2-phase cells,increasing apoptosis and the inhibiting of NF-κB activity.

Objective To label Flk-1+CD31-CD34-human mesenchymal stem cells (hMSCs) with superparamagnetic iron oxide (SPIO) and to evaluate the effect of SPIO on proliferation and neural differentiation of labeled cells. Methods hMSCs were incubated with SPIO (50 mg/L) and PLL (1.5 mg/L) overnight(12～18 hours). Both labeled and unlabeled cells went through growth curve test,Trypan blue staining and flow cytometer to evaluate the effects of SPIO on cell proliferation,cell viability and surface markers. Immunofluorescence assay was conducted for neuron and neuroglia specific cell surface markers after neural induction protocols were used. Results Cell viability of the two groups were both more than 90% for 7 days. There was no significant difference in cell viability and growth curve test between two groups. The results of flow cytometer showed that both labeled and unlabeled cells expressed CD44, CD105 and Flk-1 markers, while CD31 and CD34 were negative. After neural induction, the statistical analysis of A value for all the markers showed no significant difference between the two groups.Conclusion SPIO, as MRI cellular contrast, is safe and efficient.