Objective:To set up a eukaryotic system for high expressing human CD137 and to investigate the role of CD137 and CD137L on signals transduction of cells.Methods:The pCDNA3 plasmid containing full length of human CD137 cDNA sequence(CMV ILA SEN,CIS) and pSV2 dhfr plasmid were cotransfected into dhfr CHO cells by lipoid mediating method. The positive clone was selected with G418. Expression of CD137 on dhfr CHO cells were induced by MTX and detected by RT PCR, immunocytochemistry and flow cytometry.It's activity study was done by method of incorporating 3H TdR.Results:CD137 expressed on the surface of dhfr CHO, expression rate was 96.07%, it's activity study indicated that CD137 increase the proliferation of PBMC stimulated by anti CD3 monoclonal antibody.Conclusion:dhfr CHO cells that highly express CD137 were established. CD137 can increase the proliferation of PBMC stimulated by anti CD3 monoclonal antibody.

Objective To investigate the expression difference of microRNA -21 (miRNA-21) in patients with chronic cardiac function and analyze its clinical significance. Methods Patients with chronic heart failure (trial group,150 cases) and the subjects without chronic heart failure (control group,45 cases) were enrolled. Patients with chronic heart failure were divided into three subgroups according to NYHA: group A (heart function classⅡ,49 cases),group B (classⅢ, 51 cases) and group C (class Ⅳ, 50 cases). miRNA-21 levels were detected by qRT-PCR method. The levels of B type natriuretic peptide urea (BNP),left ventricular end diastole diameter (LVEDD) and left ventricular ejection fraction (LVEF) were determined. Results miRNA-21 expression in patients with heart failure were higher than that of control group (P < 0.01),and miRNA-21 expression in group C was higher than that of group A.Correlation analysis showed that there was a positive correlation between expression of BNP (r = 0.763,P < 0.01), LVEDD (r = 0.691,P<0.01), and the level of miRNA-21 in patients with chronic heart failure.And a negative correlation between LVEF value and the level of miRNA-21 (r = -0.918,P < 0.01). Conclusion miRNA-21 might be a potential marker for diagnosis of heart failure and could be a basis for reference about prognosis evaluation.

Background and purpose: Currently, intraoperative radiation therapy (IORT) has become the adjuvant therapy of cancer. The study was to establish the daily quality assurance (QA) program and analyze dosimetric characteristics’ long-term stability for mobile IORT accelerator. Methods:The QA program of this study included two parts:safety and functionality and energy and output. The two years’ QA datasets were acquired and analyzed to investigate the stability of energy and output. Results:All safety and functionality tests passed on a daily basis. The energy index was (0.666±0.015)mm, (0.839±0.009)mm, (0.781±0.010)mm, (0.724±0.009)mm and the output dose error was (0.511 ± 0.671)%, (0.278 ± 0.516)%, (0.368 ± 0.532)%, (0.382 ± 0.912)%for all energy, respectively. There was no signiifcant time trend in the dosimetric characteristics. Conclusion:The daily QA program is suitable for mobile IORT accelerator. The long-term stability is acceptable for IORT in clinical use.

Objective To observe the effect of 1,25(OH)_2D_3 on expression of vitamin D receptor (VDR), tumor necrosis factor α (TNF-α) and transforming growth factor β1 (TGF-β1) in rat peritoneal mesothelial cells (RPMCs) stimulated by lipopolysaccharide (LPS), so as to provide some evidence for clinical use of 1,25 (OH)_2D_3 on peritoneal dialysis-associated peritoneal inflammation. Methods RPMCs were isolated, cultured and passaged by enzymaticdisaggregation, then identified by phase contrast inverted microscope, transmission electron microscope with immunocytochemistry method. RPMCs were incubated with LPS (1,10,100 mg/L) and LPS (10 mg/L) for 2,6,12 hours, or stimulated by 1,25(OH)_2D_3 (10~(-8) mol/L, 10~(-7) mol/L, 10~(-6) mol/L) after incubated with LPS (10 mg/L) for 2 hours. RPMCs in the control group were justincubated with medium. Expression of VDR mRNA and protein was detected by RT-PCR and Western blot. In addition, ELISA was performed to investigate the changes of TNF-α and TGF-β1 in culture medium. Results Compared with control group, the expression of VDR mRNA and protein was decreased in LPS group (P<0.05). LPS could significantly induce the expression of TNF-a and TGF-β1 in RPMC (P <0.01), which could be partially reversed by different concentrations of 1,25 (OH)_2D_3 (P<0.01). Conclusions 1,25 (OH)_2D_3 can reverse the decrease of VDR mRNA and protein induced by LPS as well as the induction of TNF-a and TGF-β1 up-regulated by LPS in RPMC in a dose- and time-dependent manner. Our research provides experimental evidence of VDR ameliorating peritoneal dialysis-associated peritoneal inflammation and peritoneal fibrosis.

Objective To investigate the amlodipine or hydrochlorothiazide and valsartan combination thera-py in elderly hypertensive patients with blood pressure variability ( BPV ) and nitric oxide ( NO ) , endothelin role. Methods 552 elderly patients with hypertension were divided into the observation group and control group,276 cases in each group,the observation group was given amlodipine valsartan for treatment,while the control group was received hydrochlorothiazide combined with valsartan for treatment,the two group were treated for three months,the content of BPV,NO and plasma endothelin were analyzed.Results After treatment,the 24hSBPV,day SBPV,24hDBPV,day DBPV of the observation group significantly decreased compared with pre-treatment, the difference was statistically significant(t=20.09,15.97,9.31,9.41,all P<0.05),patients in the control group only 24hSBPV and day SBPV [(9.12 ±2.57)%,(8.43 ±1.97)%]were significantly lower than before treatment(t=12.31,9.53,all P<0.05);The day SBPV,24hDBPV,day DBPV[(7.21 ±1.34)%,(12.31 ±2.54)%,(10.59 ±2.73)%]of the ob-servation group after treatment were significantly lower than those of the control group(t=10.52,12.34,8.35,all P<0.05);The NO[ (66.12 ±23.57)%,(68.29 ±21.52)%]and endothelin levels[(34.43 ±7.97)%,(34.21 ± 7.34)%]of the two groups after treatment were significantly lower than those of before treatment(tobservation grou P=31.39,11.79,tcontrol grouP=28.31,9.35,all P<0.05).Conclusion The amlodipine or valsartan hydrochlo-rothiazide used clinically can effectively control blood pressure, significantly increase NO levels, decrease plasma endothelin levels,which is worth to be used in the clinical.

Objective To investigate the effect of sorafenib in ameliorating renal fibrosis and its possible mechanisms.Methods Rats were subjected to unilateral ureteral obstruction (UUO ) and intragastrically administered sorafenib.NRK-52E cells were treated with transforming growth factor-β1 (TGF-β1)and sorafenib. HE staining was used to visualize renal fibrosis.α-SMA and E-cadherin expressions in kidney tissue and NRK-52E cells were performed using immunofluorescence.The cell cycle of NRK-52E cells was determined by flow cytometry analysis.Smad3 and p-Smad3 protein expressions in NRK-52E cells were detected by Western blot analysis. Results HE staining showed that kidney interstitial fibrosis,tubular atrophy,and inflammatory cell infiltration in the sorafenib-treated UUO groups were significantly decreased compared with the vehicle-treated UUO group (P<0.05).Compared with those in UUO and TGF-β-stimulated NRK-52E groups,the expression of a-SMA decreased but E-cadherin expression increased in the UUO kidneys and NRK-52E cells of the sorafenib-treated groups (P<0.05).After 24 h stimulation with TGF-β1 5 ng/mL,the number of cell cycles arrested in G0/G1 phase was significantly increased and the number of cells that entered G2 ,S phase decreased (P<0 .0 5 ).Compared with that in TGF-β-stimulated NRK-52E groups, p-Smad3 decreased in the sorafenib-treated groups (P<0.05). Conclusion Our results suggest that sorafenib may be useful for the treatment of renal fibrosis through suppressing TGF-β/Smad3 signaling.

Objective To evaluate the effect of silver needle injection therapy on rat with sports muscle injury. Methods Twenty-one healthy male Wistar rats were randomly divided into the injury group (n = 3),the silver needle group (n=12) and the control group (n=3). The expressions of bFGF and GDNF in gastrocnemius muscle tendon junction were detected on 7 d ,14 d and 28 d post-injury. Results No significant difference in the appearance of the injured tissue was found in both two groups on 7 d post-injury. The appearance of the injured tissue was better in the silver needle group than that in the control group on 14 d and 28 d post-injury. The tissue was almost normal in the therapy group on 28 d post-injury; The expression of bFGF in the therapy group was higher than that in the injury control group on 7 d and 14 d post-injury (P < 0.01). The expression of bFGF markedly decreased in the therapy group compared with the control group (P < 0.01) on 28 d post-injury. The expression of GDNF in the therapy group was higher than that in the injury control group on 7 d ,14 d and 28 d post-injury (P<0.01). Conclusion The silver needle injection therapy has the therapeutic effect on sports muscle injury reparation, which can increase the expression of bFGF and GDNF efficiently.

Objective To investigate the effects of sevoflurane postconditioning on cardiomyocyte apoptosis during myocardial ischemia-repeffusion (I/R) in rats.Methods Forty-five healthy male Wistar rats weighing 250-280 g were randomly divided into 3 groups ( n =15 each):group sham operation ( group S) ; group I/R and group sevoflurane postconditioning (group Spo).The animals were anesthetized with intraperitoneal 3％ pentobarbital 45 mg/kg,tracheally intubated and mechanically ventilated.Myocardial I/R was produced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion in groups I/R and Spo.In group Spo the animals inhaled 2.5％ sevoflurane for 5 min starting from 1 min before reperfusion was started.The animals were sacrificed at the end of 120 min reperfusion.Their hearts were removed for measurement of infarct size and the area at risk and determination of apoptotic index (the number of apoptotic cells/the total number of cells) and Bcl-2 and Bax protein and mRNA expression.Results Sevoflurane postconditioning significantly reduced infarct size in group Spo as compared with group I/R.There was no significant difference in area at risk between groups I/R and Spo.Myocardial I/R significantly increased the apoptotic index,Bcl-2 and Bax protein and mRNA expression in group I/R as compared with group S.Sevoflurane postconditioning significantly decreased apoptotic index and Bax protein and mRNA expression but increased Bcl-2 protein and mRNA expression in group Spo as compared with group I/R.Conclusion Sevoflurane postconditioning attenuates myocardial I/R injury by redncing myocardial apoptosis,up-regulating Bcl-2 expression and down-regulating Bax expression.

Objective To investigate the effects of angiotensin receptor blocker (ARB)losartan on the glomerular protein expression profile of spontaneous type 2 diabetic KKAy mice by two-dimensional differential gel eleetrophoresis and MALDI-TOF mass spectrometry.Methods 8-week-old spontaneous type 2 diabetic KKAy mice were randomly divided into losartan (10 mg·kg-1·d-1 given in drinking water) treatment group and non-treatment group.Eight-week-old C57BL/6 mice were used as normal control.The glomeruli were separated by magnetic bead perfusion through thoracic aorta at age of 20 weeks,then glomerular protein was extracted.The glomerular protein expression profile was investigated by CyDyes minimal fluorescence labelling,two-dimensional differential gel electrophoresis and MALDI-TOF mass spectrometry.Results KKAy mice developed higher body weight and blood glucose,higher urinary microalbumin creatinine ratio at age of 20 weeks than C57BL/6 mice at the same age (all P＜0.05).Losartan treatment markedly reduced urinary microalbumin creatinine ratio [(539.71 ±100.23)mg/g vs (728±177.19) mg/g],attenuated mesangial expansion and the thickening of glomerular basement membrane,but had no effect on the blood glucose.By DeCyder 2-D differential analysis software,62 protein spots of differential expression were found in glomeruli between losartan treatment and non-treatment KKAy mice at age of 20 weeks.Among them,41 proteins were identified by peptide mass fingerprinting.The expressions of 28 proteins were up-regulated by losartan treatment,including glycerokinase,sulfite oxidase,glycine amidinotransferase,adenosylhomocysteinase,etc.The expressions of 13proteins were down-regulaled by losartan treatment,including 3-mercaptopyruvate sulfurtransferase,ATP synthase subunit d,60 000 heat shock protein,stress-70 protein (alternative name 75 000glucose-regulated protein,GRP75),etc.Six differcntially expressed proteins were found in glomeruli between non-treatment KKAy mice and C57BL/6 mice,and the differential expressions were suppressed by losartan treatment,including dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex,succinyl-CoA ligase (GDP-forming) subunit beta,mitochondrial,ATP synthase subunit d,GRP75,nucleoside diphosphate-linked moiety X motif 19 and seleniumbinding protein 1.Conclusions Losartan significantly reduces the urinary protein excretion rate and renal pathological lesion of spontaneous type 2 diabetic KKAy mice,and suppresses the differential expression of mitochondrial ATP synthase subunit d,GRP75,selenium-binding protein 1,etc in glomeruli.Losartan may play a renoproteetive role by reducing glomerular mitochondrial reactive oxygen species genesis and inhibiting oxidative stress.

Objective To investigate the expressions and the relationship of FEZ1 and p16 in cervical intraepithelial neoplasia (CIN).Methods The expressions of FEZ1 and p16 in 93 cases of CIN and 10 cases of normal cervical specimens were detected by immunohistochemistry.Results The positive expression rates of FEZ 1 were 90.0％ (9/10) of normal cervical specimens,67.9％ (19/28) of CIN I,36.0％ (9/25) of CIN Ⅱ,22.5％(9/40) of CIN Ⅲ.The positive expression rates of p16 were 10.0％(1/10) of normal cervical specimens,32.1％(9/28) of CIN I,76.0％(19/25) of CIN Ⅱ,92.5％(37/40) of CIN Ⅲ.There were significant differences in the positive expression rates of FEZ1 and p16 between CIN Ⅰ and CIN Ⅲ (P ＜ 0.01),there was no significant difference in the positive expression rates of FEZ1 and p16 between CIN Ⅱ and CIN Ⅲ.The expression of FEZ1 and p16 was negatively correlation in CIN (r =-0.712,P＜ 0.01).Conclusion The abnormal high expression of p16 and abnormal low expression of FEZ1 in CIN may be involved in the occurrence and development of CIN,detecting the expressions of the two indexes may be helpful for clinical diagnosis.