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 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.

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 develop an immunomagnetic bead ( IMB) assay for quantitative detection enolase ( Eno) of Candi-da albicans, and to improve the diagonosis of invasive candidiasis . Methods The immunomagnetic bead was prepared by conjuga-ting with Anti Eno of Candida albicans monoclonal antibody .HRP-conjugated goat polyclonal antibody against Candida albicans Eno was employed as detecting antibody .The performance parameters of the IMB assay including precision , specificity, linear range and limit of detection were verified by using recombinant Candida albicans Eno.Then the developed assay was applied to determine Eno levels in supernatant of pathogenic fungi cultures . Results The intra and inter-coefficient of variation was 4.54%, 5.87% and 5.26%, 8.82%at the concentration of 25 ng/mL and 5 ng/mL, respectively.The limit of detection was 0.5 ng/mL.The linear range was(0.5-50) ng/mL.The level of Eno in Candida albicans culture after incubated in 37℃for 24 h was 3.89 ng/mL and gradually in-creased to 37.89 ng/mL at 120 h.There was a positive correlation between the level of Eno and growth hyphae of Candiad albicans. There was weak cross reaction with Candida parapsilosis and no cross reaction with Candida tropicalis, Candida guilliermondii, Candida glabrata, Cryptococcus neoformans and Saccharomyces cerevisiae. Conclusion An IMB assay for quantitative detection Eno of Candida albicans was developed , which was more sensitive , rapid and reliable than previous qualitative ELISA .The IMB assay has the potential to be applied to the research in invasive candidasis .

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.

Objective： Our aim was to purity the modifier protein of glyceraldehyde-3-phosphate dehydrogenase　(G3PD)　from African green monkey Vero-E6 line. Methods：Exposure of Vero-E6 cells to medium with a reduced K concentration (3.2 mmol/L) stimulated the growth and activation of G3PD. The increase of enzyme activity was mediated by a cytosolic modifier protein that was purified using affinity and anion-exchange high-performance liquid chromatograph. Results：The apparent molecular mass of the protein was 62 kDa. Western blotting and quantiative enzyme-linked immunosorbent assay showed that the amount of modifier protein increased progressively for 2 hours in cells exposed to low-K+ medium, and then returned to the control value, a kinetic profile similar to that the modifier protein is a constituent of renal epithelial cells and accummulated transiently in the low-K+ mitogenic signal. Conclusion: We obtained a modifer protein from monkey kidney epithelial cells (Vero-E6). It could activate G3PD and cell growth.