Objective Cystatin C (CysC) is a cysteine protease inhibitor,is widely expressed in human eukaryotic cells,CysC involved in diabetic nephropathy,retinopathy,peripheral vascular disease and type 2 diabetes,insulin resistance occurs Development,as a predictor of type 2 diabetes and complications

The aim of this paper is to report the study on gene silencing efficiency of siRNA targeted against mouse VEGFR2 (siVEGFR2) in vitro mediated by polyethyleneimine (PEI) and its anti-tumor effect in vivo. CY3-labeled siRNA was compounded into PEI and transfected into MS1 cells. Confocal microscopy was used to image the subcellular distribution of siRNA in MS1 cells. Semi-quantitative RT-PCR and Western blotting were used to evaluate VEGFR2 gene silencing induced by siVEGFR2/PEI complexes. A tumor-bearing nude mice model was established to compare the anti-tumor effect after delivered by local and systemic routes. siVEGFR2/PEI complex-transfected cells exhibited much fluorescence in cytoplasm with no evidence of nuclear accumulation. The expression levels of VEGFR2 mRNA and protein in PEI-transfected cells were significantly down-regulated compared with that in blank group, the silencing efficiency were 28.2% and 23.6% respectively. The tumor sizes in mice intratumorally injected with siVEGFR2/PEI complexes (189.429 +/- 17.562 mm3) were reduced definitely compared to that in mice injected with siVEGFR2/PEI complexes via vein route (315.507 +/- 20.491 mm3), or to saline groups (365.844 +/- 20.713 mm3). The study demonstrated that PEI could effectively transfect siRNA into cells and silence the VEGFR2 gene expression. Intratumoral delivery is more suitable for non-targeted modified PEI/siRNA complexes to inhibit the tumor growth in vivo. The present data lay a solid foundation to further study on the gene silencing mechanism for PEI-medicated RNAi and its anti-tumor efficiency in vivo.

Objective To observe the influence of warm ischemia time on acquisition of rat pancreatic islets and islet function.Method Male Wistar rats were used.After heart beats stopped,the pancreases in four groups of rats were harvested,and warm ischemia time was 0,15,30 and 45 min separately.The pancrease was preserved in UW at 4℃C for 8 h,and subjected to injection of collagenase solutions.After islets were acquired,the purity,survival rate and islet activity were tested,and statistical analysis was performed.Result The number of islets obtained in 0 min group,15 min group,30 min group and 45 min group was (433 ± 41),(396 ± 38),(350 ± 31) and (66 ± 17)IEQ/one,islet viability was 94％,88％,77％ and 25％,and purity was 88％,78％,60％ and 32％,and insulin release index was 2.38 ± 0.23,2.25 ± 0.18,2.19-± 0.18 and 1.25 ± 0.12,respectively.There was no significant difference in islet number,purity,survival rate and activity 15 min group and 30 min group between 15 min group or 30 min group and 0 min group (P＞0.05).There was significant difference between 45 min group and 0 min group in islet number,purity,survival rate and activity (P＜0.05).The survival rate and purity in 45 min group were lower than the clinical standards for islet transplantation (survival rate ＞ 75％,and purity ＞ 50％).Conclusion Warm ischemia time of 15 min in non-heart-beating brain death(NHBD) rats had no effect on islet isolation and purification.Warm ischemia time within 30 min showed no significant influence on islets of NHBD rats,which can be used in islet transplantation.Warm ischemia time at 45 min showed significant influence on islets of NHBD rats,which can't be used in islet transplantation.

BACKGROUND:The use of donor rat of cardiac death inevitably experiences warm ischemia injury, so the length of warm ischemia time plays a significant role on the number and function of pancreatic islet obtained. OBJECTIVE:To investigate the effect of Exendin-4 on pancreatic islet function of donor rats with cardiac death at different heat ischemia time. METHODS:Islet cells from Wistar rats were cultured in vitro and randomly divided into three groups according to the experimental conditions:0, 30, 45 min heat ischemia groups. Each group was further assigned into two subgroups, control group was cultured for 24 hours while experimental group wad cultured with 10 nmol/L Exendin-4 for 24 hours. The number of isolated pancreatic islets was calculated with diphenylthiocarbazone staining, and the purity of the extracted islets was adjusted. The viability of the islets was examined by AO/EB staining, and insulin secretion index assay was used to detect the function of the islets. RESULTS AND CONCLUSION:With the time of heat ischemia increasing, the number, purity, viability and function of islet cells obtained were decreased. After the cells in heat ischemia 0, 30, 45 min groups were cultured with 10 nmol/L Exendin-4 for 24 hours, the number, purity and viability of isolated and purified islets were increased compared to the group without added Exendin-4. There was significant difference between experimental group and control group in 30-minute and 45-minute ischemia groups (P<0.05). Exendin-4 can protect pancreatic islet cells in donor rats with cardiac death at different heat ischemia times, reduce the apoptosis, and improve islet survival and functions. The use of Exendin-4 can be an effective pretreatment method at early ischemia phase of islet transplantation.

Diabetic mellitus induced erectile dysfunction (DIED) is a common complication of type 2 diabetes mellitus (T2DM), which seriously affects the physical and mental health of male T2DM patients. The occurrence of DIED involves a variety of pathophysiological changes such as vascular tissue, nerve, endocrine and so on. But the traditional treatment of erectile dysfunction is not ideal for DIED. Glucagon like peptide (GLP)- 1 receptor agonist has been widely used as a new drug for the treatment of diabetes, and the representative drugs are exenatide and liraglutide. Existing research shows that it can improve endothelial cell function, neuropathy and sex hormone secretion, reduce body weight and insulin resistance. Therefore, these drugs may be a new choice for patients with DIED.

Objective To investigate the electrophysiological characteristics of carpal tunnel syndrome (CTS) and CTS-associated cervical spondylotic radiculopathy (CSR),and to examine the relationship between CTS and CSR. Methods The clinical characteristics and electrodiagnostic features of 81 patients with CTS and 20 patients with both CTS and CSR (the double crush,DC) were analyzed and compared.The data were analyzed according to the severity of the deficit in median nerve conduction using electromyography. Results The 81 patients with CTS had 123 median nerves with abnormal conduction (39 cases with unilateral abnormalities and 42 cases with bilateral abnormalities).The 20 patients with DC had 31 median nerves with abnormal conduction (9 cases with unilateral abnormalities and 11 cases with bilateral abnormalities).The rate of abnormal sensory nerve conduction velocity (SCV) was 100％ in C6 and C7 level radiculopathies.The rate of abnormal distal motor latency (DML) was 92.31％ in C5 level radiculopathies.There was a statistically significant difference between CTS and DC in the rate of abnormal SCV from the middle finger to the wrist.The other electrodiagnostic data were not significantly different between the CTS and DC patients.Neurophysiological tests were used to grade CTS into categories according to the American Association of Electrodiagnostic Medicine's criteria,but there was no statistically significant difference between CTS and DC. Conclusions CSR lesions on a proximal nerve root may cause the nerve to be more susceptible to distal injury and increase the risk of CTS.The findings support the DC hypothesis,but DC on a median nerve did not result in more severe injury than a single crush.

BACKGROUND:The patients undergoing lumbar discectomy have a higher risk of recurrence. There are many different ways of reoperation, but there are few studies on spine-pelvis sagittal morphology of patients with recurrent lumbar disc herniation. OBJECTIVE:To compare the effect of discectomy and posterior lumbar interbody fusion on spine-pelvis sagittal morphology of patients with recurrent lumbar disc herniation. METHODS:Sixty-one patients of recurrent lumbar disc herniation after discectomy were divided into discectomy group (n=30) and posterior lumbar interbody fusion group (n=31) according to the re-repair method. The height of intervertebral disc, lumbar lordosis and pelvic projection angle in the two groups before and after treatment were measured and compared based on standing spine lateral X-ray images. RESULTS AND CONCLUSION: After treatment, the height of intervertebral disc, lumbar lordosis and pelvic projection angle of patients in discectomy group were not significantly changed compared with before treatment (P> 0.05). After treatment, the height of intervertebral disc, lumbar lordosis and pelvic projection angle of patients in posterior lumbar interbody fusion group were significantly increased compared with those before treatment (P< 0.05). Before treatment, there were no significant differences in the height of intervertebral disc, lumbar lordosis and pelvic projection angle between discectomy and posterior lumbar interbody fusion groups (P > 0.05).After treatment, the height of intervertebral disc, lumbar lordosis and pelvic pelvic projection angle were significantly increased in the posterior lumbar interbody fusion group compared with the discectomy group (P < 0.05). These results demonstrate that discectomy cannot significantly change the spine-pelvis sagittal morphology of patients subjected to re-operation, but compared with the discectomy treatment, posterior lumbar interbody fusion has a greater impact on spine-pelvis sagittal morphology of patients subjected to re-operation.

OBJECTIVE To understand disinfection quality in newborn Spa room,to give prevention messures against nosocomial infection.METHODS The air,UV intensity,the object surface,staff′s hands,and the using of disinfectant were detected.RESULTS The total qualified rate was 98.61% and that of the object surface was 97.22%,of staff′s hands was 98.61%,of UV intensity and indoor air was 100% and of the using disinfectant was 97.22%.CONCLUSIONS Strengthening the disinfection,finding the weak timely and taking the effective measures can prevent the occurrence of hospital infection.

Objective To detect the expression of GHR in colorectal cancer cell lines and determine whether recombinant human growth hormone can promote the proliferation of colorectal cancer cells in vitro.Methods GHR distribution was assessed by flow cytometry and immunofluorescence method in 9 colorectal cancer cell lines.The effect of recombinant human growth factor on colorectal cancer cell line proliferation was assessed by MTT method.Results Different GHR expression was determinated in 9 colorectal caner cell lines.GHR was highly expressed in HCT-8 while GHR expression could hardly be detected in LoVo.r-hGH could promote GHR(+) HCT-8 cell proliferation at 50 ng/ml and 100 ng/ml (P ＜0.05).But this effect was not dose dependent.When the neutralizing antibody was used to block GHR activity,this r-hGH dependent proliferation effect was eliminated.r-hGH could not promote GHR (-) LoVo cell proliferation (P ＞0.05).Conclusion The results demonstrates that r-hGH could promote GHR (+) tumor cell proliferation and this effect is mediated by GHR.The use of r-hGH on the colorectal cancer patients should be cautious.

OBJECTIVE In this study we explored the role of Epac1-Rap1 pathway in the acute myocardial ischemia/reperfusion injury (MIRI) in vitro and in vivo. METHODS An acute myocardial ischemia/reperfusion injury model was established by the ligation of left anterior descending coronary. Myocardial architecture, fibers and apoptosis was evaluated by the Masson trichrome staining, Sirius red staining and TUNEL assay.H9c2 cells were subjected to hypoxia for 5 h followed by 1-h reoxygen-ation in vitro.Cell viability was measured by MTT assay and cellular injury was evaluated by measuring the release of lactate dehydrogenase (LDH). Western blot, real-time PCR and immunofluorescence were used to detect the expressions of Epac1 and relative downstream molecules.RESULTS Myocardial IR-induced cardiac apoptosis and accumulation of Epac1 and Rap1 in rat IR injury model.Direct Epac activation by 8-CPT(8-(4-chlorophenylthio)-2′-O-methyl-cAMP)exacerbated cardiomyocyte death and dysfunction following hypoxia-reoxygenation(H/R),selective activation of Epac in response to H/R was evident which enriched for cytosolic/membrane proteins and mRNA. Harmacological inhibitor of Epac (ESI-09)significantly ameliorated myocardial injury with the decline of Epac expression.Epac inhibitor and agonist studies also implicated the effect of Rap1, which is downstream of Epac in this pathway. The expression of Rap1 elevated when activated by Epac agonist and was blocked by Epac inhibitor. The same result was true for myocyte CaMK-II and intracellular calcium ions activation.Moreover,ESI-09 also increased ERK1/2 phosphorylation. CONCLUSION Our study reveal that Epac1/Rap1 signaling pathway is involved in the pathogenesis of myocardial I/R injury in rats,which provides evidence on the development of therapeutic strategies target this pathway for myocardial I/R injury.