Objective:To construct the eukaryotic vector of human DC-SIGN and EGFP fusion protein,and to identify the protein in cell line COS7.Methods:RT-PCR,T-vector and pEGFP-C1 vector were used to construct the recombinant expressing plasmid encoding for the fusion protein of DC-SIGN and EGFP.COS7 cells were transfected with the plasmid.Real-time PCR and laser scanning confocal microscope were used to quantificate expression of the fusion protein and cell function of uptaking BCG.Results:DC-SIGN cDNA was successfully cloned into the eukaryotic vector pEGFP-C1.The recombinant vector was transfected into COS7,real-time PCR test showed the amount of mRNA encoding for the fusion protein was 4.52?1011 copies/ml and laser scanning confocal microscope confirmed that the cells could uptake BCG.Conclusion:We have constructed a recombinant vector expressing DC-SIGN and EGFP fusion protein.

Objective To evaluate the advantage and clinical value of javanica oil emulsion injection combined with radiotherapy for esophageal carcinoma. Method Randomized controlled trials (RCTs) concerning javanica oil emulsion injection combined with radiotherapy for esophageal carcinoma were made on electronic databases of CBM, CNKI, Wangfang, VIP and PubMed from 1990 to February 28, 2015. Quality of the included RCTs was assessed by Jadad scoring , and Meta-analyses were performed by RevMan5.3 software. Results Seven RCTs involving 575 patients were included in Meta-analysis. All studies were in low quality. The results of Meta-analyses showed that Javanica oil emulsion injection combined with radiotherapy for esophageal carcinoma, compared with control groups, could increase the recent curative effect (P < 0.000 01) and improve the quality of live (P = 0.000 8), decrease the hematologic toxicities (P = 0.03) and the incidence rate of radioaction esophagitis (P = 0.02), but not enough evidence was found to prove that it can enhance survival rate (P = 0.25). Conclusions The results of Meta-analysis indicate that javanica oil emulsion injection may have therapeutic effects on the treatment of esophageal carcinoma. Yet the effects of javanica oil emulsion injection still need to be confirmed by large multi-center randomized controlled trials.

Objective: To develop 5-FU multiple emulsion entrapped into thermo-sensitive gel (5-FU-DEG) and detect the ab-sorption and transportation in Caco-2 cell monolayer model. Methods:The 5-FU multiple emulsion was prepared by a two-step emulsif-ying method. Poloxamer 407 (P407) was used as the thermo-sensitive material and sodium alginate (SA) was used as the bioadhesive material for the preparation of 5-FU-DEG. Caco-2 cell monolayer model was used to investigate the transportation and absorption of 5-FU. Results:5-FU-DEG gelled at the ambient temperature and turned into liquid below 10℃ The apparent permeability coefficient (Papp) of 5-FU-DEG was 1.47 ±0.11 ×10 -5(cm·s-1), which was about 6 times higher than that of 5-FU water solution(2.39 ± 0.21 ×10 -6 cm·s-1)(P<0.01). The cellular uptake rate of 5-FU-DEG was (17.1 ±0.24) %, which was 3.9 times greater than that of 5-FU water solution (4. 41 ± 0. 23%)(P<0. 01). Conclusion:5-FU-DEG can efficiently enhance the transportation and ab-sorption of drug in rectal site by using micro-emulsion technology combined with thermo-sensitive technology, which can be an effective rectal delivery system for 5-FU to treat rectal cancer.

Objective To investigate the effect of berberine on glucose absorption and mRNA expression of 11 β-hydroxysteroid dehydrogenase type 1 ( 11β-HSD1 ) in insulin-resistant HepG2 cell model.Methods HepG2 cells were incubated with high-concentration insulin for 24 hours to build insulin-resistant cell model.In order to evaluate the cells for insulin resistance,the cells were cultured with different concentrations of insulin for 24 hours.The insulin-resistant cells were treated with different concentrations of berberine and insulin for 24 hours and the non insulin-resistant cells were used as a control.The glucose concentration in culture medium was detected by the method of glucose oxidase-peroxidase (GOD-POD).According to the glucose concentrations in blank medium and those in the medium of culturing cells 24 hours later,the rate of absorpting glucose by the cells was calculated.The mRNA expression of 11β-HSD1 in the insulin-resistant cells was detected by the reverse transcription polymerase chain reaction (RT-PCR).Results Incubated with 10-7mol/L insulin for 24 h,the insulin-resistant cell model had been built.The rate of glucose absorption of the model cell treated with high concentration berberine ( 10 μmol/L) was significantly improved [(42.53 ±1.99)％ vs (28.16±1.99)％,t =12.9457,P ＜0.01].High-concentration berberine showed a strong synergy with insulin on glucose absorption of the model cells.As the cells became resistant to insulin,the mRNA expression of 11β-HSD1 increased significantly compared to non insulin-resistant cells( relative expression quantity was (4.60 ±0.96 vs 0.67 ±0.42,t =4.9476,P ＜0.05 ).While the mRNA expression of 11β-HSD1 reduced in the insulin-resistant cells after treated with high-concentration berberine,and the relative expression quantity was not significantly different with non insulin-resistant cells ( 1.12 ±0.35 vs 0.67 ±0.42,P ＞0.05).However,low-concentration ( 1 μmol/L) of berberine had not the same role.Conclusions It is concluded that one of the acting mechanism of berberine improving the insulin sensitivity may be that the mRNA expression of 11β-HSD1 is downregulated in the insulin-resist-ant liver cell model depending on concentration.

Objective To investigate the current status of self-efficacy and care ability among stroke patients′ family caregivers and explore the correlation between them. Method A total of 79 stroke patients and family caregivers were engaged in the investigation by demographical data questionnaires, general self-efficacy scale and questionnaire for assessing the comprehensive care-giving ability of stroke caregivers. Results The score on self-efficacy of the family caregiver was (27.16 ± 5.64) and the total score on care ability was (90.01 ± 16.57). There was a positive correlation between self-efficacy and care ability of family caregivers(P<0.05). Conclusions The self-efficacy and care ability of stroke patient′s family caregivers are at middle level. Their self-efficacy is positively correlated with their care ability. The nurses can improve the ability of daily caring of family caregivers by enhancing their self-efficacy.

Most main oilfields in China have already entered a "double high" development stage (high water cut, high recovery degree). To further enhance oil recovery in reservoirs after polymer flooding (RAPFs), an efficient activator formulation for promoting metabolism of endogenous microorganism was studied by aerogenic experiments, physical simulation experiments, electron microscopy scanning and pyrophosphate sequencing. Results show that the activator could activate the endogenous microorganisms in the injected water and make the pressurized gas reach 2 MPa after 60 d static culture of the activator in a high pressure vessel. The oil recovery efficiency of natural core physical simulation flooding can be improved by more than 3.0% (OOIP) in RAPFs when injected 0.35 PV activator with 1.8% mass concentration, and a lot of growth and reproduction of activated endogenous microorganism in the core was observed by electron microscopy scanning. Field trial with 1 injector and 4 producers was carried out in the east of south II block of Sa Nan in December 2011. By monitoring four effective production wells, changes of carbon isotope δ13C (PDB) content of methane and carbon dioxide were -45 per thousand to -54 per thousand and 7 per thousand to 12 per thousand. Compared with east II of Sa Nan block, the oil amount increased by 35.9%, water cut stabled at 94%. The incremental oil was 5 957 t during the three and a half years, which provides an alternative approach for further improving oil recovery in similar reservoirs.
Carbon Dioxide ; chemistry ; Carbon Isotopes ; analysis ; China ; Diphosphates ; chemistry ; Methane ; chemistry ; Oil and Gas Fields ; microbiology ; Polymers ; Water ; Water Microbiology

Objective To evaluate the association between 8 single nucleotide polymorphisms (SNPs)of vascular endothelial growth factor (VEGF ) gene and the risk of Alzheimer’s disease (AD ) in Shaanxi Han population.Methods We examined the potential association between AD and 8 SNPs of VEGF gene using the MassARRAY system.The participants enrolled in this study included 214 patients with AD and 249 healthy controls from Shaanxi Han population.SPSS16.0 and Haploview 4.2 were employed to analyze differences in genotypes, alleles and haplotypes between the two groups.Results The results showed that rs3025039 (3’UTR)were significantly associated with AD (P<0.05).Greater frequency of rs3025039 T allele (P=0.008,OR=1.527,95%CI=1.116-2.088)was found in AD subjects.Furthermore,strong linkage disequilibrium (LD)was observed in 2 locks (block1:rs699947-rs1570360-rs2010963;block 2:rs3024997-rs3024998-rs3025006)(D’>0.9).There were no significant haplotypes in block 1 and block 2 (P=0.034)found between the patients and controls.Conclusion These findings point to the role for VEGF gene polymorphisms (rs3025039)in AD of a Shaanxi Han population. Individuals with T allele of rs3025039 may be at a higher risk for AD.

Objective To investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on calcium homeostasis and calcium channels of PC12 cells induced by ischemic and hypoxia and its mechanisms. Methods PC12 cells at logarithmic phase were collected, and they were divided into recombined lentiviral infection group [infected by lentivirus containing HSP70 and green fluorescent protein (GFP) fluorescin gene], lentivirus control group (infected by lentivirus containing GFP without HSP70 gene) and non-infection group. PC12 cells were subjected ischemia/hypoxia for 4, 8, 12, 24 hours, and the cell activity was determined by methylthiazolyl tetrazolium (MTT) assay test inorder to determine the best time for ischemia/hypoxia. The mRNA expressions of HSP70, muscle/endoplasmic reticulum Ca2+-ATP isoforms (SERCA2a, SERCA2b), ryanodine receptor 2 (RyR2), and inositol 1,4,5-triphosphate receptor 1 (IP3R1) were determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and the protein expressions of HSP70, SERCA, and IP3R were determined by Western Blot at 8 hours after ischemic/hypoxia. Flow cytometry was used to determine the levels of intracellular reactive oxygen (ROS) and intracellular Ca2+ ([Ca2+]i). Results With the prolongation of time of ischemia/hypoxia, the cell viability in all groups showed an increase followed by a weakening, and peaked at 8 hours. The cell viability at 8 hours in lentiviral infection group was significantly higher than that of the non-infection group and lentivirus control group [A value (×10-2): 20.3±2.2 vs. 14.1±2.1, 15.0±1.6, both P < 0.01], the mRNA and protein expressions of HSP70 and SERCA in lentiviral infection group were significantly increased [HSP70 mRNA (2-ΔΔCt ): 0.785±0.018 vs. 0.428±0.019, 0.423±0.023; HSP70 protein (gray value): 2.72±0.20 vs. 1.56±0.36, 1.63±0.41; SERCA2a mRNA (2-ΔΔCt ): 0.971±0.037 vs. 0.367±0.014, 0.347±0.012; SERCA2b mRNA (2-ΔΔCt ): 8.869±0.162 vs. 3.015±0.091, 2.941±0.091; SERCA protein (gray value): 2.84±0.18 vs. 1.48±0.26, 1.52±0.29], and IP3R2 mRNA and protein expressions were significantly declined [IP3R2 mRNA (2-ΔΔCt ): 0.183±0.020 vs. 0.439±0.020, 0.433±0.040; IP3R2 protein (gray value): 1.15±0.12 vs. 1.91±0.20, 1.83±0.19], with statistically significant differences (all P < 0.01); no significant difference in RyR mRNA was found [2-ΔΔCt (×10-3): 1.97±0.63 vs. 2.02±0.22, 2.01±0.09, both P > 0.05]; the relative fluorescence intensity of ROS and [Ca2+]i in lentiviral infection group was significantly reduced (ROS: 30.54±1.23 vs. 58.03±1.97, 57.72±2.35; [Ca2+]i: 34.50±2.05 vs. 48.20±3.02, 46.80±2.75, all P < 0.01]. Conclusion Exogenous HSP70 can maintain calcium homeostasis in the endoplasmic reticulum of PC12 cells, affect the Ca2+ channel protein regulated by calcium channel IP3R and calcium pump SERCA, which may cause hypoxia/ischemia intracellular injury.

Objective To identify the pathogen distribution and antibiotics resistance of blood stream infection(BSI) in the pediatric surgery intensive care unit(PSICU).Methods The clinical data of 138 pediatric patients diagnosed with BSI from January 2011 to December 2015 were collected in PSICU,and the distribution of pathogens and drug resistance were retrospectively analyzed.Results The incidence of BSI was 3.88‰(138/35.524)in the five years,the majority of the BSI cases occurred under one year old,and the mortality was 13.77%(19/138).A total of 179 strains were isolated from blood samples of 138 patients,of which gram-positive bacteria,gram-negative bacteria and fungi accounted for 60.89%(109/179),22.91%(41/179)and 16.20%(29/179)respectively.The most common gram-positive bacteria was coagulase-negative staphylococcus (84/179,46.93%).The predominant gram-negative bacteria were Acinetobacter baumannii(15/179,8.38%),Klebsiella pneumonia(12/179,6.70%) and Escherichia coli(6/179,3.35%).The rate of carbapenems-resistant Acinetobacter baumannii increased continuously in the study period.Non-albicans Candida was the most common fungi (14/179,7.82%).The resistance rate of multi-drug resistant strains to carbapenems significantly increased.Conclusion The incidence of BSI in PSICU increases,and the mortality in children younger than one year is high.Better understanding of distribution of BSI pathogen could provide more effective antibiotic prescription.

Objective To explore the prognostic value of biomarkers in type 2 diabetic patients with acute myocardial infarction (AMI), this study was to investigate the associations between the neutrophil to lymphocyte ratio (NLR), the Global Registry of Acute Coronary Events (GRACE) score and in-hospital mortality. MethodsSeven hundred and seven consecutive AMI patients were divided into diabetic group (DM-AMI group), impaired glucose tolerance group (IGT-AMI group), and normal glycemic group (NGT-AMI group). The laboratory and clinical characteristics were assessed retrospectively from the medical records. The NLR and GRACE score were calculated. Results In AMI patients, the DM-AMI group had significantly higher NLR and GRACE scores compared with those from the IGT-AMI group and NGT-AMI group (P<0.01 or P<0.05). In DM-AMI group, the NLR and GRACE score were considerably elevated in the elderly DM-AMI group compared with their younger counterparts (both P<0.01). Furthermore, the NLR was considerably higher in the high-risk group than those in both the low- and medium-risk groups based on the GRACE score (both P<0.01). The NLR was positively correlated with the GRACE score in DM-AMI group(r=0.425, P<0.01). The NLR level and GRACE score were higher in the death group than those in surviving patients (both P<0.01). The optimal cut-off levels of 9.36 for NLR and 166 for GRACE score seem to predicte death in-hospital. Based on the receiver operating characteristic curve, when to predict death in-hospital, the best cutoff value of NLR was 9.36 (sensitivity 80.8%, specificity 69.6%; area under curve 0.787), and the best cutoff value of GRACE score was 166 (sensitivity 76.9%, specificity 76.4%; area under curve 0.778). Conclusion An elevated NLR is a potential predictor of in-hospital mortality in type 2 diabetic patients with AMI, which could help clinicians indentify high-risk patients and determine appropriate treatment strategies. <英文关鍵词>>=Neutrophil to lymphocyte ratio; In-hospital mortality; Acute myocardial infarction; Diabetes mellitus, type 2