Objective To study the expression of the fused proteasome activator REGγ(11S regulator complex gamma subunit) using gene recombination technology and to further study the interaction between REGγ and casein kinase-2 interacting protein-1(CKIP-1)in vitro.Methods Firstly, the full length cDNA fragment of REGγ was amplified through PCR using the plasmid pCMV-Myc-REGγ as template and subcloned into the prokaryotic expression vector pGEX-4T-2 before being transformed into E.coli BL21 cells. The protein expression was induced by isopropyl-β-D-thiogalactoside(IPTG) .Secondly, the protein expression was monitored by SDS-PAGE and Western blotting after ultrasonication. Finally, the GST Pull-down assay was performed to investigate the interaction between REGγ and CKIP-1 in vitro.Results The prokaryotic expression construct pGEX-4T-2-REGγ was generated successfully and confirmed by DNA sequencing. Expression analysis showed that the GST-REGγ protein was easily expressed and isolated mainly in the lysate supernatant after sonication and centrifugation. The GST Pull-down assay revealed the strong mutual interaction between REGγ and CKIP-1 in vitro.Conclusion The proteasome activator REGγ could interact with the negative regulator of osteoblastogenesis CKIP-1 in vitro and the current study has shed light on further investigations of their physiological relevance.

Objective To investigate expression of transforming growth factor-?1(TGF-?1)and connective tissue growth factor(CTGF) protein in renal tissues,and detect the levels of urinary TGF-?1 and CTGF in rats with diabetic nephropathy(DN).To observe the influence of losartan on expression of the two protein in renal tissue and excretion in urine.Methods Wistar rats were treated by intravenous injection of streptozotocin after right nephrectomy to induce DN rat model.The DN rats were randomly divided into two groups:DN experimental group and losartan treated group.The expression of TGF-?1 and CTGF in renal tissue were determined with immunohistochemical staining,urinary TGF-?1 and CTGF were assayed by enzyme-linked immunosorbent assay(ELISA) at 6,12 weeks respectively.Results Compared with losartan treated group,urinary protein excretion and the protein expression of TGF-?1 and CTGF significantly increased(P

This study was purposed to analyze the efficiency of platelet transfusion and to explore factors influencing platelet transfusion efficiency. 727 times of platelets transfusion in 254 patients in The Third Xiangya Hospital from September 2010 to May 2011 were analyzed retrospectively. Moreover, according to symptoms, times of platelet transfusion, blood types and splenomegaly, the corrected count of increment (CCI) and percentage of platelet recovery (PPR) were calculated for evaluation of platelet transfusion efficiency. The results showed that there were 456 effective transfusions out of 727 transfusions (62.72). Among them, the therapeutic effect of platelet transfusion for patients with acute blood loss anemia and chronic systemic diseases was relatively obvious, specially for chronic renal disease, the effective efficiency of them was 94.12. The patients with splenomegaly showed a significant impact on platelet transfusion efficiency (41.07). Analysis found that the frequency of platelet transfusion negatively correlated with transfusion efficiency. It is concluded that the transfusion frequency and splenomegaly are factors influencing the transfusion efficiency.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Middle Aged ; Platelet Count ; Platelet Transfusion ; methods ; Retrospective Studies ; Treatment Outcome ; Young Adult

SCID mouse-human leukemia model is an important and useful tool for study on proliferation, differentiation and modulation of leukemic cells. In this article, the establishment of the model, advances in research and application in studies of pathogenesis, cell biology, clinical diagnosis, therapy and assessment of prognosis of leukemia patients are reviewed. The limitations of the model are also commented.
Animals ; Disease Models, Animal ; Humans ; Leukemia ; therapy ; Mice ; Mice, SCID ; Research Design

OBJECTIVETo investigate the relationship between the changes of the copy numbers of mtDNA in peripheral blood mono-nucle- ar cell(PBMC) and the disordered of antioxidant capacity of hepatocellular carcinoma (HCC) patients.

METHODSThe Ficoll Hypaque method was used to isolate the PBMC from blood specimens. The ND1 gene of the mitochondrial was amplified by real-time PCR; meantime β-actin was served as a quantitative standard marker; the difference of mtDNA copy number in PBMC was compared between HCC and healthy control group. The level of reactive oxygen species (ROS) in PBMC was determined by flow cytometry. The change of total antioxidant capacity (T- AOC) of plasma was detected by the biochemistry examination.

RESULTSThe copy numbers of ND1 gene in PBMC of HCC was 73% that of the healthy control group,which suggested a decrease of the copy numbers of mtDNA in HCC. The levels of ROS of PBMC in HCC was (417. 82 ± 110.62) and (301.82 ± 75.54) in control group, which showed that the levels of ROS of PBMC in HCC were significant higher than that in control group (P < 0.01).Plasma T-AOC in HCC was (1.30 ± 0.85), and (3.20 ± 1.62) in control. The T-AOC of plasma of HCC was significantly lower than in control group (P < 0.01).

CONCLUSIONThere was a certain relationship between the decrease of the copy numbers of mtDNA and the disordered antioxidant capacity in hepatocellular carcinoma, which may be associated with the development of hepatocellular carcinoma.

Actins ; Antioxidants ; metabolism ; Carcinoma, Hepatocellular ; blood ; genetics ; Case-Control Studies ; DNA Copy Number Variations ; DNA, Mitochondrial ; genetics ; Humans ; Leukocytes, Mononuclear ; metabolism ; Liver Neoplasms ; blood ; genetics ; Reactive Oxygen Species ; metabolism

so higher in diabetic patients 4 h after the meal (all P<0. 05). Positive correlation existed between serum triglycerides and white blood cell counting, neutrophils, and high-sensitivity C-reactive protein(r were between 0.268 and 0.548, all P<0.05).

Visually impaired people face many inconveniences because of the loss of vision. Therefore, scientists are trying to design various guidance systems for improving the lives of the blind. Based on sensory substitution, auditory guidance has become an interesting topic in the field of biomedical engineering. In this paper, we made a state-of-technique review of the auditory guidance system. Although there have been many technical challenges, the auditory guidance system would be a useful alternative for the visually impaired people.
Acoustics ; Auditory Perception ; Biomedical Engineering ; Equipment Design ; Humans ; Sensory Aids ; Software ; Sound Localization ; Visual Perception ; Visually Impaired Persons ; rehabilitation

OBJECTIVETo explore the transmission routes, genotypes/subtypes distribution and genetic character of HCV in HIV/HCV co-infected and HCV mono-infected individuals in Guangdong Province.

METHODSReverse transcription (RT) nested PCR was performed to amplify the HCV NS5B gene region from 95 HIV/HCV co-infected and 99 HCV mono-infected individuals lived in Guangdong province. The PCR products were then sequenced for HCV subtyping. Genetic analysis was done by MEGA4 software.

RESULTS(1) HIV/HCV co-infected individuals infected HCV mostly through injection drug use (IDU, 78.9%), the HCV subtypes were identified as 6a (53.7%), 3a (17.9%), 1b (15.8%), 3b (11.6%) and 1a (1.0%) respectively, the genetic distance within subtype 1b was longer than those within other subtypes, the predominant HCV subtype in HIV/HCV co-infected individuals infected through IDU was 6a (60.0%). (2) HCV mono-infected individuals infected HCV mostly through blood or blood products transfusions (80.8%), the HCV subtypes were identified as 1b (67.7%), 6a (17.2%), 3a (6.1%), 2a (5.0%), 3b (2.0%), 4a (1.0%) and 5a (1.0%) respectively, the genetic distance within subtype 1b was also longer than those within other subtypes, the predominant HCV subtype in HCV mono-infected individuals infected through blood or blood products transfusions was 1b (76.2%).

CONCLUSIONThe diversity of HCV subtypes in HIV/HCV co-infected and HCV mono-infected individuals in Guangdong Province was high, both the major transmission route and HCV subtype between HIV/HCV co-infected individuals and HCV mono-infected individuals were different.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; China ; epidemiology ; Coinfection ; virology ; Female ; Genotype ; HIV ; HIV Infections ; epidemiology ; virology ; Hepacivirus ; classification ; genetics ; Hepatitis C ; epidemiology ; virology ; Humans ; Male ; Middle Aged ; Phylogeny ; Young Adult

The aim of this study was to investigate the apoptosis-inducing effect of artemisinin derivative SM1044 on Kasumi-1 cells and its possible mechanism. Kasumi-1 cells were treated with different concentrations of SM1044, the cell viability was evaluated by MTT assay. Cell apoptosis and cell cycle progression were assessed by using flow cytometry with Annexin-V/PI double staining and flow cytometry with PI staining respectively. The expression of apoptosis-related proteins caspase 3, PARP and the fusion protein AML1-ETO were detected by Western blot. The results indicated that SM1044 inhibited cell growth of Kasumi-1 cells in time- and dose-dependent manners. After exposure of Kasumi-1 cells to 1 µmol/L SM1044 for 24 hours, the cell viability was decreased to 50%. IC(50) of SM1044 to Kasumi-1 cells at 48 hours was 0.17 ± 0.067 µmol/L. SM1044 induced cell apoptosis in a caspase-dependent manner, and the apoptotic rate of Kasumi-1 cells increased as SM1044 concentration increased. Flow cytometry with PI staining revealed that SM1044 induced cell cycle arrest, and the proportion of cells in G(0)/G(1) phase increased from 58.33 ± 4.46% to 71.75 ± 2.24% after exposure to 5 µmol/L SM1044 for 24 hours. Western blot showed that SM1044 increased the expression of apoptosis-related proteins cPARP and cleaved caspase 3 and also degraded the AML1-ETO fusion protein. It is concluded that SM1044 can inhibit the proliferation of Kasumi-1 cells, induce cell apoptosis which may be related to the increased level of cleaved PARP and cleaved caspase 3. SM1044 can also induce cell arrest in G(0)/G(1) phase. As the fusion protein AML1-ETO degrades obviously, it can be the potential target of SM1044 in Kasumi-1 cells.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Artemisinins ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Leukemia, Myeloid, Acute ; pathology

OBJECTIVETo evaluate the feasibility of using iron oxide nanoparticles as gene vector and the effect of magnetic field on efficiency of transfection.

METHODSIron oxide nanoparticles were prepared by alkaline precipitation of divalent and trivalent iron chloride. The surface of iron oxide nanoparticles was modified by self-assembled poly-L-lysine to form particle complexes (IONP-PLL). Transfection was determined by delivering reporter gene, PGL2-control encoding luciferase, to different cell lines using IONP-PLL as vector. The effect of magnetic field on efficiency of transfection was determined using Nd-Fe-B permanent magnet.

RESULTSForeign gene could be delivered to various cell lines by IONP-PLL and expressed with high efficiency, but the transfection efficiency and time course varied in the different cell lines studied. Magnetic field could enhance the efficiency of transfection by 5 - 10 fold.

CONCLUSIONIONP-PLL can be used as a novel non-viral gene vector in vitro, which offers a basis for gene delivery in vivo.

Animals ; COS Cells ; Ferric Compounds ; administration & dosage ; Genetic Vectors ; Magnetics ; Polylysine ; administration & dosage ; Transfection ; methods