Objective To determine the diagnostic significance of serum Cystatin C (Cys C),neutrophil gelatinase-associated lipocalin (sNGAL),urinary kidney injury molecule1 (uKIM-1) and urinary interleukin-18 (uIL-18) in patients with septic acute renal injury.Methods A total of 62 adult patients with sepsis admitted to the Intensive Care Unit over 24 hours in the Tianjin Hospital between August 2012 and March 2013 were divided into acute renal injury group (n =39) and non-acute renal injury group (n =23) according RIFLE diagnostic criteria.Measurements of Cys C,sNGAL,uKIM-1 and uIL-18 were performed.The diagnostic values of the biomarkers were assessed by comparing their levels between the patients of different groups.Results The levels of Cys C (2.27 ± 0.93) μg/L,sNGAL (275.04 ±79.37) μg/L,uKIM-1 (2.52 ± 1.06) μg/L in acute renal injury group were higher than Cys C (1.19 ± 0.77) μg/L,sNGAL (137.51 ± 99.33) μg/L,uKIM-1 (1.27 ± 0.62) μg/L in non-acute renal injury group (P ＜ 0.05).There was no significant difference in uIL-18 between the acute renal injury group (374.87 ± 70.23) ng/L and the non-acute renal injury group (354.09 ± 66.42) ng/L (P ＞ 0.05).The area under ROC curve for diagnostic values of serum Cystatin C,sNGAL and uKIM-1 in acute renal injury were0.84 (0.74-0.95),0.90 (0.79-1.00),0.87 (0.78-0.96).Conclusions CysC,sNGAL and UKIM-1 might be valid as diagnostic biomarkers of septic acute renal injury.

Objective: To explore change of heart rate turbulence (HRT) in patients with coronary artery disease (CAD) and its predictive value. Methods: A total of 58 CAD patients were regard as CAD group. The 50 normal persons with negative CAG were regard as normal control group. All subjects received 24h dynamic ECG, CAG and echocardiography examination. Left ventricular ejection fraction (LVEF) and HRT indexes were compared between two groups. Results: Compared with normal control group, there was significant rise in turbulence onset [TO, (-1.34±1.09)% vs. (2.32±0.60)%] and significant reduction in turbulence slope [TS, (11.14±2.27) ms/RR vs. (3.44±0.60)ms/RR] in CAD group, P<0.01 both; in CAD group, along with coronary artery lesion aggravated (single-, double-, multi-vessel coronary disease), there was significant rise in TO [(1.35±0.52)% vs. (2.56±0.83)% vs. (3.01±1.62)%] and significant reduction in TS [(6.49±1.79) ms/RR vs. (3.33±1.02) ms/RR vs. (1.66±0.30) ms/RR], the difference was significant in two-two comparison among all groups, P<0.05 or <0.01; compared with CAD without cardiac event group, there was significant rise in TO [(1.68±0.61)% vs. (2.24±0.24)%], and significant reduction in TS [(5.87±1.22) ms/RR vs. (1.65±0.23) ms/RR] and LVEF[(66.18±2.64)% vs. (56.36±3.34)%] in CAD with cardiac event group, P<0.01 all. Conclusion: Weakened HRT phenomenon exists in CAD patients. HRT can be treated as an index evaluating severity of CAD patients’ condition and an effective predicting index for recurrent cardiac events in CAD patients.

Objective To investigate the expression of miRNA-16 (miR-16) in breast cancer patients and its effect on the proliferation and migration of breast cancer cells. Methods Polymerase chain reaction (PCR) was used to detect the expression of miR-16 in 30 breast cancer patients. miR-16 mimics was transfected to MDA-MB-231 and MCF-7 cell, and CCK-8 as well as Transwell assay was applied to detect the effect of miR-16 on cell proliferation and migration of MDA-MB-231 and MCF-7 cells. Results Among 30 breast cancer patients, the expression of miR-16 was down-regulated in 23 cases. Cell proliferation and migration of MDA-MB-231 and MCF-7 cells were inhibited significantly after transfection of miR-16 mimics. Conclusion miR-16 is down-regulated in breast cancer patients. miR-16 inhibits significantly the cell proliferation and migration of breast cancer cells in vitro.

Objective To establish a stable model of intraluminal thread approach in making focal cerebral ischemia/reperfusion modelin rabbit.Methods Forty New Zealand white rabbits were randomly divided into three groups:A,B,and C.Angiography of ICA wasperformed in group A(n=5). Control group B(n=5) was sham-operated.In group C(n=30),the focal cerebral ischemia/reperfusionmodel was made by inserting a wire ( d=0.45 mm ) into MCA through ICA and CCA directly and pulling it out 3 hours later . MRI andneurological deficit score were used to evaluate the effectiveness of models.Results Angiography could show the course and bifurcation of ICA clearly.22 models were successful.The pathological changes were shown on MRI.The neurological deficit score (reperfusion 1 h) were 2.8?0.45.Conclusion The focal cerebral ischemia/reperfusion model made by inserting a wire(d=0.45 mm) into MCA through ICA and CCA directly and pulling it out 3 hours later was stable and fit for imaging research.

Yeast surface display is a new technology developed in recent years,which displays protein of interest on the surface of yeast cell.It can be used to display complex eukaryotic proteins requiring post-translational processing including glycosylation and efficient folding for functional activity. The basis of this technology, its development,methods of screening,its applications and perspective are mainly described in this paper.

Objective To construct the eukaryotic expression vector coding HCV gene E2 fused with His-Tag, and to express fused protein in CHO cells for investigating the function of HCV envelope protein E2. Methods The gene encoding HCV envelope protein E2 was amplified from pBRTM/HCV1-3011, a plasmid containing the cDNA of HCV's ORF, by polymerase chain reaction (PCR) method and cloned into the vector pET28(a) containing His-Tag to obtain the fused HCV envelope protein E2 gene fused with His-Tag. The fused gene was cloned into pcDNA3.1 to construct the recombinant plasmid pcDNA3.1-His-E2, which will express the E2 protein, fused with His tag. This recombinant plasmid was transfected into CHO cells by Lipofactamine 2000 reagent. The fused protein was identified by indirect immunofluorescence (IIF) and Western-blot (WB) methods. Result The positive results were obtained when the fused protein of HCV E2 with His-Tag were identified by IIF and WB methods. Conclusion The eukaryotic expression vector pcDNA3.1-His-E2 was constructed successfully and the fused proteins were expressed in cells.

Objective To investigate the state of oxidative stress in the subjects with different levels of serum uric acid and to explore the cause of endothelial dysfunction induced by hyperuricemia. Methods Male subjects with normal serum uric acid or hyperuricemia were enrolled in this study. According to the levels of serum uric acid,all the partieipante were divided into five groups. Every group consisted of about fifty subjects. Plasma malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase(GSH-Px) were determined. In the mean time plasma nitric oxide(NO) ,plasminogen activator inhihitor-1(PAI-1), endothelin-1 (ET-1) and other biochemical variables were also determined. Results When the serum uric acid level was more than 380 μmol/L, the levels of plasma MDA,SOD,GSH-Px, PAI-1 and ET-1 became higher, while the level NO became lower(P<0.05). However SOD and GSH-Px became much lower when the serum uric acid level was more than 420 μmol/L. Multivariate stepwise regressive analysis showed that PAI-1 was positively related to MDA,UA,HOMA-IR and TG, but negatively related to SOD and NO(t =-3.64 - 6. 08,P < 0.05). ET-1 was positively related to MDA, UA, HOMA-IR and negatively related to NO, GSH-Px and SOD (t = - 4.75 - 6.35,P < 0.05 ). Conclusions It is indicated that oxidative stress became much obvious when the serum uric acid level was more than 380 μmol/L . Oxidative stress,high serum uric acid level and insulin resistance may result in endothelial dysfunction.

Objecflve To evaluate the performance of two rapid and low-cost metheds(MTT test,and rosazurin mierotitre assay)for the detection of resistance to first-line drugs in Mycobacterium tuberculosis.Methods sixty-four Myeobaeterium tuberculosis clinical isohtes were tested by the MTT test and the rosazuxin microtitre assay(REMA)respectively,and the results were compared with those obtained with the absolute concentration method on L(o)wenstein Jensen medium.Results The MTT test and the resazurin microtitre assay showed a good agreement compared with the absolute concentration method for all first-line drugs tested.The sensitibity,specificity and accuracy of the MTT test were 94.8%,96.0%,95.3%,for RFP;93.8%,93.8%,93.8% for INH;92.9%,96.O%,95.3% for EMB,90.6%,87.5%,89.1% for SM,respectively.The sensitivity,specificity and accuracy of the resazurin microtitre assay were 92.3%,96.0%,93.8%,for RFP;90.6%,90.6%,90.6% for INH;92.9%,94.0%,93.8% for EMB,87.5%,87.5%,87.5% for SM,respectively.The Kappa value of the MTT test and the absolute concentration method for the detection of resistance to RFP,INH,EMB,SM were 0.857,0.831,0.714,0.792.respeedvely;The Kappa value of the regazurin mierotitre assay and the absolute concentration method for the detection of resistance to RFP,INH,EMB,SM were 0.871,0.826,0.826,0.750,respectively.The Kappa value of the MTT test and the resazurin microtitre assay for the detection of resistance to RFP,INH,EMB,SM wefe 0.889,0.875.0.787,0.844,respectively.Conclusions Both MTT test and the resazurin microtitre assays are simple,rapid,low-cost and sensitive for rapid detection of resistance to first-line drugs.They could be promising methods for susceptibility assay of the first-line antituberculosis drugs in low-resource countries.

Objective To investigate the diagnosis value of a new tumor marker CK18-3A9 in gastric cancer patients. Methods The serum level of CK18-3A9 in 350 gastric cancer patients, 150 gastritis patients and 500 healthy controls was detected with chemoluminescence, the diagnosis efficacy between the serum CK18-3A9 and CA72-4 was compared. Results The sensitivity, specificity of CK18-3A9 were 46.29 % and 96.92 %, but the sensitivity, specificity of CA72-4 were 26.00 % and 93.23 %, respectively. The differences were significant (P ＜0.001). Conclusion The serum level of CK18-3A9 maybe a new auxiliary diagnosis marker for gastric cancer.

Objective To analyze the clinical characteristics of five Sj(o)gren's syndrome (SS) patients with severe ostalgia. Methods Five SS patients from the endocrinology department of our hospital were retrospectively analyzed. Results All the patients were female, with 9.5 year average course of disease, obvious ostalgia, muscle weakness and complaint, limitation of activity, and different degree of exocrine gland impairment. All the patients had different degree of acidosis, disorders of calcium and phosphorus metabolism, and dyspoiesis of calcitriol. Among them, three patients were found to have hyperinsulinemia and muscle pathological changes detected by the biopsy. Conclusions System factors must be investigated when patients present the bone symptom. At the same time, muscle problem should not be neglected and the patients should be treated as an entirety.