Objective: To investigate the regulatory effect of HIV-1 Tat on mitotic centromere-associated kinesin (MCAK) expression and the related mechanism. Methods: Tat-expression TE671 cell model and purified prokaryotically expressed Tat protein were used to verify the down-regulated expression of MCAK using Northern blotting and Western blotting analysis. Various DNA fragments in the promoter region of the MCAK gene were amplified with PCR, linked to the luciferase reporter plasmid, and then transferred into TE671 cells. Luciferase activity analysis was performed to measure the promoter activity of various DNA fragments, so as to determine the active promoter region of MCAK gene. Moreover, HIV-1 Tat protein was co-incubated with TE671 cells, and the promoter activity was detected to analyze the modulating effect of Tat on promoter activity in vitro. Results: The results of Northern blotting and Western blotting analysis indicated that the mRNA and protein levels of MCAK were strongly decreased by Tat protein. The core region of MCAK promoter was located in -399∼+1 bp region, and the promoter activity was strikingly inhibited by Tat protein. Conclusion HIV-1 Tat has a marked inhibitory effect on MCAK expression, which might be related to the decreased promoter activity caused by Tat protein.

Objective: To observe the expression of DNA dependent protein kinase catalytic subunit (DNA-PKcs) in the hepatoma tissues and mouse liver tissues of different developmental stages, so as to understand the role of DNA-PKcs in cell proliferation and tumorigenesis. Methods: Immunohistochemistry and Western blotting assay were used to observe the protein expression of DNA-PKcs in liver tissues and hepatoma tissues. The siRNA technique was used to silence the expression of DNA-PKcs in the HepG2 cells; cell proliferation assay and tumor transplantation test in nude mice were performed to evaluate the changes of the proliferation ability and tumorigenesis. Western blotting assay was also conducted to examine the expression of proliferation related proteins p-GSK3β and c-myc. Results: DNA-PKcs expression decreased in the liver tissues with the decrease of cell proliferation ability during the development of mice, and the expression level of DNA-PKcs was weak in liver tissues of adult mouse; but the DNA-PKcs protein level in the hepatoma tissues was significantly elevated (P<0.01). Cell growth curve showed that the proliferation of HepG2 cells was significantly decreased after suppression of DNA-PKcs with siRNA(P< 0.01), accompanied by a suppressed tumorigenesis ability. The expression of signal pathway related protein p-GSK3β and c-myc was inhibited after DNA-PKcs silencing in HepG2 cells(P<0.01). Conclusion: DNA-PKcs expression level is closely related to the proliferation ability of liver cells. Overexpression of DNA-PKcs may participate in the development and progression of hepatoma through mediating cell proliferation via the Wnt/GSK/c-myc related signal pathway.

Objective To study the effect of siRNA targeting survivin gene on the apoptosis of malignant melanoma cell line A375. Methods The eucaryotic expression vector of pU-survivin-siRNA was constructed and transfected into the A375 cells by electroporation. The protein expression of survivin was examined by Western blotting, and cell apoptosis by flow cytometry. Results The transfection of pU-sur-vivin-siRNA significantly down-regulated survivin expression ( 0.24 ?0.02 in the transfected group versus 0.98 ?0.21 in the control group ) in A375 cells, and promoted cell apoptosis ( 83% in the transfected group versus 28% in the control group, P

Adenosine Triphosphate ; pharmacology ; Animals ; Burns ; metabolism ; Calcium ; metabolism ; Female ; Male ; Mitochondria, Heart ; metabolism ; Rats

Objective To understand the current cognition status of stroke rehabilitation nursing knowledge in young nurses of the department of neurology.Methods Self-designed questionnaires were used for investigating 273 young nurses between 20 and 30 years old of the department of neurology of 4 tertiary hospitals in Zhengzhou.Results The current status of stroke rehabilitation nursing knowledge of the nurses surveyed was generally at a low level.There were no significant differences among nurses with different education degrees,professional titles and whether their relatives suffered from stroke.However,there were significant differences among nurses with different professional lives and whether the department carried out rehabilitation care.Conclusions Nursing educators and managers need to find a new way to improve the level of stroke rehabilitation nursing knowledge of the young nurses.

Objective To discuss the pathogenesis,diagnosis and treatment of delayed traumatic intracerebral hematoma.Methods The clinic data of traumatic delayed intracranial hematoma patients in this hospital were retro- spectively analyzed.According to clinic observation and CT re-examination,47 cases were diagnosed as delayed trau- matic intracranial hematoma(45 cases by operative treatment,and the other 2 by conservative treatment).Results There were 21 cases of recovery,10 cases of slight disability,8 cases of severe disability,8 cases of death.The total mortality rate was 17 %.Conclusion Brain contusion,subarachnoid hemorrhage and skull base fracture were impor- tant factors of DTICH.Fine-observation and prompt CT re-examination offered excellent results for DTICH.

Objective To investigate the effect of escharectomy on rats' pulmonary NF-?B activation and the expression of pulmonary proinflammatory cytokines in early stage of burn injury.Method Wistar rats were randomly divided into three groups:group A(control group),group B(postburn without escharectomy),group C(escharectomy at early stage of burn injury).Thermal-injuried rats underwent 35% TBSA full-thickness burns. Activation of pulmonary NF-?B at 12 hours and 24 hours postburn was tested by electrophoretic mobility shift assay (EMSA),and at the same time expressions of pulmonary TNF-?mRNA were measured by reverse transcription polymerase chain reaction(RT-PCR)and release of pulmonary TNF-?were assayed by enzyme-linked immunosorbent assay(ELISA).Results Compared with control group,activity of pulmonary NF-?B in group B was markedly increased,reached(19.56?1.36)?10~4 A at 12 hours and(15.23?1.94)?10~4 A at 24 hours,which was higher than that in group A[(4.36?0.38)?10~4 A,P

The potential of Rhizobium sp. N613 to produce the exopolysaccharide (REPS) was studied in this paper. Using an orthogonal design in a flask-shaker culture system, the fermentation medium and conditions of synthesizing REPS were optimized. Based on these results, the fermentation kinetic parameters were obtained in the batch fermentation with a 10L fermentor. The REPS yield of 11.31g/L was achieved by metabolic regulation during 40 h fed-batch fermentation. Transplanted tumor models of sarcoma 180 in mice were used to evaluate the anti-tumor effect. The result of anti-tumor activities showed that inhibition rate was 53.40%, when dose of REPS was 5mg/kg. These results indicate that REPS has the following properties: the short duration of fermentation, the high yield, the low cost, the effective immunocompetence and thickening. Thus, REPS has the value of development and application.

The most toxic DNA damage is DNA double strand breaks (DSBs), which are mainly repaired by non-homologous end joining (NHEJ). DNA-dependent protein kinase (DNA-PK) belongs to phosphatidylinositol-3-kinase-related protein kinase family (PIKK) and plays a key role in NHEJ. DNA-PK is overexpressed in a variety of cancer cells and is related to the occurrence, development and drug resistance of malignant tumors. In this article, the representative DNA-PK inhibitors with anticancer effects are reviewed, in order to provide a reference to discovery novel DNA-PK inhibitors.

BACKGROUND: Dialysis-related amyloid may occur during long-term dialysis for patients with uraemia, of which the main evocator is β_2-microglobulin (β_22M); therefore, how to eliminate 132M from blood is always the focus of research. OBJECTIVE: To observe ability of removal of β_2-microglobulin (β_2M) from serum using two kinds of polyethersulfone (PES) membrane materials with various degrees of sulfonation.METHODS: These materials were incubated in radio-labeled β_2M (~(125)Ⅰ-β_2M) solution and human serum respectively at appointed time at 37 ℃, and then the amounts of ~(125)Ⅰ-β_2M and serumβ_2M adsorbed by materials were measured by radio immunoassay. RESULTS AND CONCLUSION: In the ~(125)Ⅰ-β_2M system, amounts of ~(125)Ⅰ-β_2M adsorbed by the materials decreased in the following sequence PES with high degree of sulfonation > PES with low degree of sulfonation > PES, whatever the source of PES was. In the serum system, amounts of β_2M adsorbed reached maximums at 30 minutes and the final adsorptions decreased in sequence of PES with high degree of sulfonation > PES with low degree of sulfonaUon > PES. Sulfonated PES removed β_2M more than PES did and the adsorption of β_2M increases with the increase in the degree of sulfonation. Its ability to remove significant amount of β_2M may result in less β_2M available for incorporation into amyloid. The use of sulfonated PES membranes may lessen the likelihood of development of dialysis-related amyloidosis, which remains a major source of morbidity for patients treated with long-term hemodialysis.