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.

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 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 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 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.

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 investigate the effect of 12-lipoxygenase(12-LO) on the p27Kip1 expression in diabetic glomeruli. Methods Mesangial cells were exposed to 12-LO product 12 (S)-HETE (10-7 mmol/L) with or without p38 MAPK (p38) inhibitor (SB203580, 1 μmol/L) for 24 hours. Rats fed with high fat diet received low dose streptozotoein (ST-Z, 35 mg/kg, IP injection) to develop type 2 diabetes and were divided into 2 groups: low dose STZ, low dose STZ+12-LO inhibitor cinnamyl-3,4-dihydroxy-α-cynanocinnamate (CDC, 8 mg/kg) treatment. Rats fed with regular chow were divided into two groups: controls, CDC treatment. The rats received injection of CDC or vehicle subcutaneously in the hind leg. CDC or vehicle injection was performed three times weekly on alternate days. All the rats were sacrificed after 4 weeks, Wild type and 12-LO knockout C57BL/6 mice were divided into 4 groups: wild type control, 12-LO knockout, STZ-induced wild type type 1 diabetes and STZ-induced 12-LO knockout type 1 diabetes. All the mice were sacrificed after 16 weeks. Urine, blood, kidney cortical tissue and isolated glomeruli by sieving method were collected at the end of study respectively. Western blot and immunohistochemistry for target protein were performed respectively. Results Inhibition of p38 activation could significantly reduce p27Kip1 expression induced by 12 (S)-HETE in mesangial cells (P<0.01). Increased glomerular volume, microalbuminuria, elevated glomeluli p38 activation, p27Kip1 expresssion in type 2 diabetic glomeruli was decreased after CDC treatment (P<0.01). Compared with wild type diabetic mice, glomerular p38 activation, p27Kip1 exprcsssion and extracellular matrix accumulation in the 12-LO knockout diabetic mice were significantly decreased (P <0.01, respectively). Conclusions 12-LO induces p27kipl expression via p38 pathway in diabetic glomeruli.

The results of RT-PCR and immunohistochemistry showed that the expressions of vascular endothelial growth factor (VEGF) and its receptor ( flk-1 ) in the renal cortex of insulin-resistant rats during the phase of normal blood glucose were significantly increased, which were decreased by telmisartan. The result suggests that telmisartan may ease kidney damage via decreasing VEGF and flk-1 expressions.

ObjectiveTo observe the therapeutic effects of Shenxiong glucose injection in the patients with chronic kidney diseases.Methods Seventy-eight patients with chronic kidney disease were given intravenous glucose injection 200 ml,qd,for 2 weeks.The patients who were complicated with diabetes would be given insulin 2 U/100 ml.Before and after Shenxiong injection treatment,24 h urinary protein,blood urea nitrogen ( BUN),creatinine ( Cr),hemoglobin ( Hb),albumin ( ALB),hematocrit ( HCT),fiber fibrinogen (FIB),cholesterol (TC) and triglyceride (TG) were measured and compared.Results After 1 course of Shenxiong treatment,the serum BUN,Cr,FIB,24 h urinary protein,Hb and HCT ( [ 15.70 ± 3.62 ] mmol/L vs.[6.74 ± 1.56 ] mmol/L,[ 564 ± 65 ] μmol/Lvs.[ 189 ± 43 ] μmol/L,[ 0.08 ± 0.01 ] g/L vs.[ 0.04 ± 0.01 ] g/L,[7.96 ±3.45]g vs.[3.60 ± 1.92]g,[83.6 ±10.5]g/L vs.[79.5 ±8.7]g/L,[0.43 ±0.0] vs.[0.39 ±0.06 ] were decreased,and ALB ( 28.7 ± 8.6) vs.( 36.8 ± 6.2) was increased.The difference was statistically significant (t =3.1 1,2.98,3.04,2.82,2.02,2.23,P＜0.01 for all).TC and TG were not changed much after the treatment.ConclusionShenxiong injection can improve the kidney function,lower Fibrinogen and urine protein,which may effectively slow down the progress of chronic kidney disease and improve the life quality.