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 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 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 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 effects of ketogenic amino acid ( KAA) replacement diet on insulin resistance in mice fed with high fat diet(HFD) and to analyze the possible mechanism. Methods C57BL mice were fed with a control diet, HFD, and KAA-fortified HFD(HFDKAAR)from the age of 8 weeks, and 8 weeks after HFD initiation, the HFD-fed mice were divided into two groups:one group of mice were fed the same HFD, the other group were fed HFDKAAR ( HFD→HFDKAAR ) . The metabolic evaluations were performed at the end of 16 weeks. Blood glucose levels were measured at 0, 15, 30, 60, 90, and 120 min after the injection of glucose ( 1 g/kg BW intraperitoneal glucose tolerance test, ipGTT) . The insulin,β-hydroxybutyrate, and acetoacetate levels in the plasma were measured via ELISA. The insulin resistance index ( IRI) and area under curve ( AUC) were calculated. The expression of hepatic LKB1 ( liver kinase B1 ) , AMP-activated protein kinase ( AMPK ) , and mTOR ( Mammalian target of rapamycin ) protein, and mcp-1 mRNA were measured by western blot and real-time PCR respectively. Results HFD-fed group of mice displayed significantly heavier body weight,heavier intra-abdominal fat weight, and significant deterioration of glucose tolerance at the end of 16 weeks in addition to higher insulin levels( all P<0. 05), HFDKAAR-fed mice exhibited significantly ameliorated high fat diet-induced obesity and glucose intolerance compared to the HFD-fed mice, which was associated with decreased insulin levels, IRI, AUC, and mcp-1 mRNA expression (all P<0. 05). HFD suppressed hepatic LKB1 and AMPK phosphorylation expression, and increased mTOR phosphorylation levels compared to the control diet-fed mice(all P<0. 05). In contrast, treatment with the HFDKAAR diet increased LKB1and p-AMPK expression, which was associated with suppressed p-mTOR levels compared to the HFD-fed mice(all P<0. 05). Conclusion KAA may ameliorate high fat diet-induced obesity, glucose intolerance, via normalizing the hepatic LKB1-AMPK-mTOR nutritional signal passageway. KAA replacement diet seems to be a potential nutritional intervention for the treatment for patients with metabolic defects, such as obesity, glucose intolerance, as well as metabolic syndrome.

Objective:To investigate the effect of continuous blood purification (CBP) on inflammatory factors and immune function in patients with sepsis,in order to provide reference for clinical treatment of sepsis.Methods:A total of 76 patients with severe sepsis,who were treated in ICU of Daqing Oilfield General Hospital from June 2014 to December 2016,were selected.The patients were divided into conventional group (n=36) and CBP group (n=40) according to the received treatment method.The patients in the conventional group were treated with targeted therapy and life support,while the patients in the CBP group were treated with CBP on the basis of targeted therapy.The levels of interleukin-6 (IL-6),interleukin-10 (IL-10),tumor necrosis factor-α (TNF-α) and C reactive protein (CRP) of the patients in the two groups were detected and compared before treatment and after 3d of treatment.The CD3+,CD4+,CD8+T cell and the CD4+/CD8+ ratio before treatment and after 3 d of treatment were calculated and compared between the two groups.Results:There was no significant difference in the level of serum inflammatory factors between the two groups before treatment (P＞0.05).The levels of serum inflammatory factors in the two groups were significantly decreased after 3 d of treatment,and the levels of IL-6,TNF-α and CRP in the CBP group were significantly lower than those in the conventional group (P＜0.05).There was no significant difference in immune function indexes between the conventional group and the CBP group before treatment (P＞0.05).After 3d of treatment,CD3+,CD4+ and CD4+/CD8+ ratio were significantly increased in the two groups,and the CD4+ and CD4+/CD8+ ratio of the CBP group were sig-nificantly higher than those of the conventional group (P＜0.05).Conclusion:The application of CBP in the treatment of patients with sep-sis can obviously improve the expression of inflammatory factors of patients,and can improve the immune function and promote the re-covery of immune function.

By stratified random sampling method, 10 000 residents were investigated with the questionnaire in Fangzhuang community of Beijing, 9497 valid questionnaires were collected with a return rate of 94. 97%. The results showed that two main purposes for patients to use the community health services were to seek consultation (29. 8% ) and to get prescription (28. 2% ). The three main reasons for residents to choose community health services were the convenience, less medical expenses and more clear explanation from doctors, followed by the personal trust in the doctors.

Objective To investigate the expression of osteopontin(OPN) and monocyte chemoattractant protein-1(MCP-1) and the effect of Irbersartan on them in diet-hypercholesterolemia rats. Methods Wistar rats were divided into three groups: normal control rats(C), cholesterol fed rats(H) and cholesterol fed rats treated with Irbesartan(50 mg ? kg -1 ? d -1 ). Twelve weeks later, we measured the 24 hours total urine protein, creatinine clearance and total serum cholesterol, LDL- cholesterol, HDL- cholesterol and triglycerides. Kidney pathology was observed. Immunohistochemistry was used to analyse the expression of OPN, MCP-1, ED1 +and their relationship. Results (1) Total serum cholesterol, LDL-cholesterol level and 24 h total urine protein in H group rats were higher than that in C group rats, there was no significant difference between two groups in HDL-cholesterol and triglycerides.(2) Compared with C group rats , the expression of OPN and MCP-1 increased in cortical tubular epithelium (2.34?0.25 vs 0.49?0.11; 1.93?0.21 vs 0.49?0.11, P