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.

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

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

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.

OBJECTIVETo investigate the result of hip arthroplasty for failed internal fixation of femoral neck fractures.

METHODSFrom June 2007 to January 2014, 29 cases who underwent hip arthroplasty for failed of internal fixation of femoral neck fractures were reviewed. There were 12 males and 17 females. The mean age was 60.3 years (ranged 43 to 83 years) at the time of the fracture. Left hip was in 16 cases, right hip was in 13 cases. The average interval from fracture to arthroplasty was 23.3 months (ranged, 3 to 48 months).

RESULTSAll of 29 cases were performed total hip arthroplasty. There were 20 cases of cementless cup,7 cases of cementless cup with bone graft, 2 cases of cemented cup with bone graft; 13 cases of cementless stem, 16 cases of cemented stem. There were no complications occurred such as intraoperative fracture of the greater trochanter. The average operative time was (115 ± 38) minutes,the mean intraoperative blood loss was (420 ± 175) ml, the average postoperative drainage volume (240 ± 119) ml, intraoperative blood transfusion was (200 ± 220) ml, intraoperative fluid volume was (2,200 ± 400) ml, the average postoperative blood transfusion was (300 ± 200) ml. There was 1 case get postoperative dislocation. All patients were followed up for 14.7 months in average (ranged, 5 to 24 months). There was no revision for mechanical failure. Harris Hip Score significantly was improved from 51.1 ± 7.5 before the conversion to 88.5 ± 6.4 points at the final follow-up.

CONCLUSIONThe effect of the hip replacement for patients with failed internal fixation of femoral neck fractures was confirmed. This method can shorten the time on the bed and reduce the complications. It benefits the patients earlier functional recovery, but it must control operation indication. The long term efficacy is necessary to further observation.

Adult ; Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; methods ; Female ; Femoral Neck Fractures ; surgery ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Treatment Failure

BACKGROUND: The dynamic changes of pneumocyte apoptosis and aspartate-specific cysteine proteases-3 (caspase-3) expression in lung tissue of rats during the process of lung ischemia/reperfusion (I/R) injury and the possible action mechanisms remain unclear.OBJECTIVE: This study was to observe the dynamic changes of pneumocyte apoptosis and caspase-3 expression in the rat lung tissue during the process of lung I/R injury, and to analyze the role of pneumocyte apoptosis and the possible action mechanism.DESIGN: A randomized controlled animal experiment.SETTING: Emergency Center, First Hospital, Nanjing Medical University.MATERIALS: This study was carried out in the Animal Laboratory of the First Hospital of Nanjing Medcial University and Nanjing Center for Radioimmunity between April 2006 and September 2006. Twenty-eight male healthy SD rats of clean grade, with body weight of 250 to 350 g, aged 49 to 76 days, were provided by the Experimental Animal Center of Nanjing Medical University. The involved rats were randomized into experimental group and control group, with 14 rats in each.METHODS: ①Experimental intervention: Rats in the experimental group were created into models of lung I/R injury according to the method of Eppinger et al. They were occluded for 45 minutes at the porta of lung (no systolic and diastolic reactions in lung tissue being considered as successful occlusion), and then they were reperfused (recovery of systolic and diastolic function being considered as successful reperfusion); After that, lung tissues were harvested at 3 and 6 hours after lung I/R injury, 7 rats at each time point. Each rat in the control group was subjected to a thoracotony only, but lung tissues were isolated at the same time point by the same method. ②Experimental evaluation: Apoptotic cells in the lung tissue were detected with a flow cytometer by Annexin-V-PI staining, and apoptosis rate was calculated. Caspase-3 expression in the lung tissue was observed by immunohistochemical method and image analysis. Wet to dry weight ratio(W/D) of lung tissue of rats in the two groups was calculated; the number of injured pulmonary alveoli at I/R 3 hours/that at I/R 6 hours was calculated for quantitative evaluation of injured lung tissue; Patho-morphological changes of lung tissue were observed by haematoxylin & eosin staining under an optical microscope.MAIN OUTCOME MEASURES: ①Pneumocyte apoptosis rate and caspase-3 expression in the lung tissue. ②W/D of lung tissue and quantitative evaluation of injured lung tissue. ③Patho-morphological changes of lung tissue.RESULTS: Twenty-eight rats were involved in the final analysis, without deletion. ①Pneumocyte apoptosis rates in the experimental group at I/R 3 and 6 hours were significantly increased as compared with control group (P＜0.01). In the experimental group, pneumocyte apoptosis rate was decreased a little at I/R 6 hours than at I/R 3 hours (P＜0.05). ②Caspase-3 expression in the lung tissue of rats of experimental group reached its top at I/R 3 hours, and was decreased a little at I/R 6 hours. At each time point, caspase-3 expression in the experimental group was increased as compared with control group (P＜0.01). ③In the experimental group, the number of injured pulmonary alveoli at I/R 3 hours/that at I/R 6 hours and W/D ratios of lung tissues were significantly increased as compared with control group (P＜0.01). In the experimental group, two ratios at I/R 6 hours were higher than those at I/R 3 hours (P＜0.05).④In the experimental group, the structure of pulmonary alveoli was destructed, collapsed and disappeared; lots of inflammatory cell infiltration was found; Patho-morphological changes of injured lung tissue at I/R 6 hours were severer than those at I/R 3 hours. No obvious changes were found in the control group.CONCLUSION: At the early stage of lung I/R injury, the alteration of caspase-3 maybe activate pneumocyte apoptosis and induce the apoptosis of lung tissue, and thereby leads to lung injury.

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.