Objective To construct the endothelial cell model with overexpressed human protein kinase C ?2(PKC?2) after high glucose inducement in order to study the function of human PKC?2. Methods The PRKCB1 gene was amplified from pMD18-T-PRKCB1 plasmid and then directly cloned into shuttle plasmid pDC315 to construct shuttle plasmid. Then the recombinant shuttle plasmid and adenovirus genomic plasmid pBHGlox△E1,3Cre were cotransfected into 293 cells to construct recombinant adenovirus Ad-PRKCB1. The virus titer was calculated by TCID50. The Ad-PRKCB1 was verified by immunocytochemistry and RT-PCR. Ad-PRKCB1 was transfected into human umbilical vein endothelial cells (HUVECs) followed by the treatment of high glucose (25 mmol/L) for 96 h, and then the cells were observed by laser scanning confocal microscopy (LSCM). Results Restrictive endonuclease digestion and PCR result showed that the target gene was correctly cloned into shuttle plasmid. Cytopathic effect (CPE) was observed under inverted microscope through homologous recombination. The virus titer was 7.9?109 IU/ml. Immunocytochemical staining and RT-PCR indicated the expression of human PKC?2 in the transfected HUVECs. The translocation was observed under LSCM. Conclusion A recombinant adenovirus vector with human PKC?2 and the endothelial cell model with human PKC?2 overexpression by high glucose are constructed successfully.

Objective To construct the eukaryotic expression vector of human PRKCB1 containing enhanced green fluorescence protein gene and transfer into human umbilical vein endothelial cells(HUVECs),then identify activity of expression protein.Methods The pReceiver-M29-PRKCB1 eukaryotic expression plasmid was constructed by frame amplified from pMD18-T-PRKCB1 plasmid.Then the recombinant plasmids were identified by enzyme analysis and DNA sequencing.According to optimized conditions,the eukaryotic expression plasmids were transfered into HUVECs and observed under fluorescence microscope.After that,transfection efficiency was calculated under random vision.The plasma membrane/cytosol ratio of fluorescence was calculated under confocal microscope.The translocation was identified.Results The gene sequence was completely consistent with that reported in GenBank.The enhanced green fluorescence protein was observed in HUVECs after 48 hours.Transfection efficiency was 18.6%?1.6%.The translocation was observed.Conclusion The eukaryotic expression plasmid is successfullyconstructed and transfered into HUVECs,the translocation was identified.It is a potential tool for screening HUVECs stably expressing human protein kinase C ?2 and isolating protein complex.

Animals ; Hindlimb ; blood supply ; Ischemic Preconditioning ; Lung ; metabolism ; Male ; P-Selectin ; metabolism ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism

BACKGROUND: Rhodiola rosco L acts on anti-aging,anti-fatigue and improving immunity in human body and improves angina pectoris.OBJECTIVE: To study the effect of rhodiola root on heart hemodynamics and myocardial contractility and probe into its mechanism.DESIGN: Observational and controlled based on the experimental animals.SETTING: Department of physiology in a medical university.MATERIALS: The experiment was performed in Laboratory of Cardiac Vessels of Department of Physiology of Guangxi Medical University,from January to April 2001.Thirty New Zealand white rabbits of either sex,weighted varied from 1.9 to 2.3 kg,were randomized into two groups,named as the control(physiological saline 4 mL/kg) and rhodiola sacra SH Fu (nuodikang) group(0.5 g/4 mL · kg-1),15 rabbits in each group. A hundred common wild frogs were employed,weighted varied from 120 to 160 g,randomized into 4 groups, named as the control group of physiological saline,nuodikang of small dosage group(blood concentration 209 mg/L),nuodikang of moderate dosage group(blood concentration 418 mg/L) and nuodikang of large dosage group(blood concentration 627 mg/L),25 frogs in each group.METHODS: ms2000 multiple medium biological signal analysis system was applied,by Straub method,frog heart perfusion specimen in vitro was prepared and the cardiac activity was assayed. The relevant indexes of ventricle activity in rabbits were determined with hemodynamics.MAIN OUTCOME MEASURES: Comparison of hemodynamics changes in rabbits of two groups and the comparison of effects of nuodikang at various dosages on frog heart work in vitro.RESULTS: Myocardial contractility in every nuodikang group was strengthened remarkably compared with the control(F = 8.939, P<0.01 ), the myocardial contractility in nuodikang groups at the dosages of 209 mg/L,418 mg/L and 627 mg/L was in the tendency of increasing,strengthened by 52. 3%,70. 1% and 86.4% respectively. But the significant difference was not apparent in the comparison among experimental groups(P>0.05). The significant difference in frog heart rates among groups was not apparent(P>0.05). Concerning to hemodynamics in rabbits,in nuodikang groups,the maximum increasing rate of pressure in the left ventricle was increased by 22.1% (F = 6.259,P<0.05),the mean pressure of artery was decreased by 8.2% (F = 5.688, P<0.05) . The significant difference in other indexes was not apparent in the comparison between the control and nuodikang groups.CONCLUSION: Nuodikang strengthens myocardial contractility and speeds up myocardial contraction, but does not present remarkable influence on myocardial dilation; simultaneously it reduces the mean pressure of artery.

In order to evaluate the changes in T cell subsets after acute irradiation injury, and investigate the mechanism of deficiency of cellular immunity in acute irradiation syndrome, Balb/c mice were exposed to a single total body irradiation of 5Gy gamma rays from a 60 Co source. On days 10, 28 and 120, the amounts of CD4 + and CD8 + subsets in spleen cells were evaluated by flow cytometry (FCM), and expression of IL 4 gene in ConA stimulated spleen cells was evaluated by reverse transcription linked polymerase chain reaction, in which ?-actin gene was used as an internal control. It was found that after irradiation, the ratio of CD4 + /CD8 + became significantly higher than that of the controls, and then the ratio lowered gradually until day 120. However, it still did not return to the level of the controls. On day 10, expression of IL 4 gene at mRNA level increased significantly. The results indicated that CD8 + subset was more radiosensitive than CD4 + , and the pattern of cytokine seeretion shifted to that of Th2 soon after irradiation.

Abstract: Objective （ ） To investigate the current status of medical radiation protection in medical and health institutions MHI Methods - （ ） in Tibet Autonomous Region. Sixty one MHIs in seven prefectures cities of Tibet Autonomous Region were selected as the study subjects by stratified random sampling. The radiological protection equipment and personal protective ， equipment were investigated and the quality control of radiological equipment and radiation protection monitoring in Results ， radiological workplace were monitored. There were 368 radiation workers in 61 MHI institutions accounting for 4.8% （ ） ∶ ， 368/7 701 of the total number of radiation workers. The ratio of male to female was 2 1 and the average was six people/ institution. The quantity of radiation monitoring equipment and personal protective equipment at all levels of MHI was less than ， 1.00 sets/person. Among them tertiary MHI had the lowest number of personal protective equipment configurations. The （ monitoring qualified rates of radiation equipment quality control and radiation protection in radiation workplace were 73.3% 88/ ） （ ）， 120 and 95.8% 115/120 respectively. The lowes tmonitoring qualified rate of radiation equipment quality control was 55.2% （ ） ， （ ， ）Conclusion 32/58 with digital radiography and the second was computed tomography 84.8% 28/33 . The monitoring ， qualified rate of radiation protection in MHI workplaces at all levels in Tibet Autonomous Region is high. However radiation monitoring equipment and personal protective equipment should be increased.

Objective:To investigate the influence of clinical outcomes and the risk factors of poor prognosis for preoperative nutritional risk in gastric cancer patients.Methods:A prospective study was performed in 140 patients with gastric cancer and the nutritional risk screening 2002 (NRS 2002) was done.The influence of preoperative nutritional risk on postoperative complications,hospital stay,ICU stay,hospital expenses,60 days readmission and mortality was analyzed,and the risk factors of perioperative complication were identified by univariate and multivariate analysis.Results:The significant difference (NRS 2002 ≥ 3 group vs.NRS 2002 ＜ 3 group) was observed in the rates of overall postoperative complications,pulmonary infection,overall hospital stay and postoperative hospital stay (P ＜ 0.05).The rates of anastomotic fistula,pleural effusion,60 days readmission,60 days mortality and hospitalization expenses in NRS 2002 ≥ 3 group were higher than that of NRS 2002 ＜ 3 group,but there were no differences between the two groups (P ＞ 0.05).By univariate and multivariate analysis,preoperative high cholesterol levels and preoperative nutritional risk are the risk factors of poor prognosis and postoperative complications.Conclusion:Preoperative high cholesterol levels and preoperative nutritional risk are independent risk factors of postoperative complications.

Congenital Hyperinsulinism ; diagnosis ; Humans ; Infant, Newborn ; Male

OBJECTIVETo investigate the progressive motility, (PR), total motility (progressive + non-progressive motility, PR + NP), and acrosin activity of sperm from normal and infertile men at different time points after sperm activation.

METHODSBased on the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen and the results of modified Papanicolaou staining, we divided the semen samples into groups A (normal, n = 28), B (oligoasthenoteratospermia, n = 30), and C (asthenoteratospermia, n = 32). At 1, 24, and 48 hours after sperm activation, we detected sperm PR and PR + NP by CASA and chemical colorimetry, and determined sperm acrosin activity using the modified Kennedy method.

RESULTSSperm PR and PR + NP were significantly decreased in all the three groups at 1-24 hours and even more significantly at 24-48 hours after sperm activation as compared with the baseline (P < 0.05). Sperm acrosin activity showed remarkable reduction in group A (P = 0. 013) , even more significant at 1-24 hours than at 24-48 hours after sperm activation, but not in groups B and C (P = 0.519 and 0.979).

CONCLUSIONSperm PR, PR + NP, and acrosin activity are all decreased with the extension of time after sperm activation, each in a specific manner. Examination of sperm acrosin activity should be applied as a routine tool in the assessment of male fertility.

Acrosin ; metabolism ; Asthenozoospermia ; metabolism ; physiopathology ; Biomarkers ; metabolism ; Humans ; Infertility, Male ; metabolism ; physiopathology ; Male ; Semen ; Sperm Motility ; physiology ; Spermatozoa ; metabolism ; physiology ; Time Factors

Flavonoid naringin is widely distributed in various types of plants and is an important component of herbal Drynaria. In previous studies, Drynaria has been demonstrated to have inhibitory effects on inflammatory responses and bone destruction and exert anabolic effects on bone, has been widely used in the clinical treatment. Naringin, was in the stage of experimental yet. The experimental results have confirmed that naringin suppressed inflammation including arthritis by lowering the expression of inflammatory cytokines, and the mechanism can be explained as reducing the expression of NF-κB. Naringin has been shown to increase osteoblast proliferation by increasing the expression of BMP-2, inhibit osteoclast activity by reducing the expression of RANKL. In animal experimental, naringin was useful for osteoporosis, and the mechanisms are in-depth studies. Research in the field of traditional Chinese medicine and orthopedics, naringin as a explicit material structure in the components of Drynaria, has been concerned about the experimental studies, it is not only prosperity the development of traditional Chinese medicine research,but also ready for clinical studies anti-inflammatory and bone effects of naringin in the future.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Bone and Bones ; drug effects ; Flavanones ; pharmacology ; Humans ; Medicine, Chinese Traditional ; Osteoblasts ; drug effects ; Osteoclasts ; drug effects ; Osteoporosis ; drug therapy ; Polypodiaceae ; chemistry