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 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.

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 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.

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

OBJECTIVETo establish a miniature pig model of mandibular osteoradionecrosis (ORN).

METHODSA single dose of irradiation (IR25 Gy, 28 Gy) was delivered via an electronic linear accelerator to the right mandible of 24 miniature pigs by utilizing three dimensional conformal radiotherapy (3D-CRT). The first molar of the mandible was extracted 2 months after radiation. Mandibular ORN was diagnosed through gross observation, X-ray film, CT, and histopathologic examination.

RESULTSAt 3-4 months after radiation, all animals suffered from mandibular ORN after dental extraction. The symptoms of ORN were more severe in the 28 Gy group than in the 25 Gy group.

CONCLUSIONA miniature pig model of mandibular ORN can be established through 25 Gy irradiation + dental extraction.

Animals ; Mandible ; Mandibular Diseases ; pathology ; Molar ; Osteoradionecrosis ; pathology ; Particle Accelerators ; Radiation Injuries, Experimental ; pathology ; Radiotherapy, Conformal ; adverse effects ; Swine ; Swine, Miniature ; Tooth Extraction

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.

[Objects]To isolate and identify the pathogen of the large outbreak of dengue in Guangdong province in 2014. To understand the origin and the phylogenetic characteristics of the isolates ,and provide scientific foundation for the surveillance and prevention of dengue fever.[Methods]Collected the patient serum samples over all the Guangdong province during the 2014 outbreakperiod,isolated and identified the virus from these samples. Amplified complete E gene and complete genome with certain primers and sequenced all the products. Then the Phylogenetic ,Bayesian phylogeography and mutations analysis were carried.[Results]40 DENV-1 strains were isolated and identified. 40 complete E gene sequences and 6 complete genome sequences of DENV-1 were obtained. Phylogenetic analysis with E gene sequences revealed that the 40 isolates were classified into two genotypes including 16 genotypeⅠ(Asia)and 24 genotypeⅤ(America/Africa). 14 genotypeⅠisolates were clustered closest with isolates from Guangdong province(2013)and Sigapore(2013)which share the nucletide identities of 99.6% ~ 99.9%,other two genotypeⅠisolates were clustered with strains from Malaysia (2013) and both share the nucletide identities of 99.7%;24 genotypeⅤisolates were all classified in one clade with striains from Bangladesh(2009),China(2009)and Bhutan(2013)which share nucletide identities of 99.0%-99.9%. Further analysis with six complete genome sequences showed that five isolates were clustered closest with strains isolated from Guangdong province(2013)share the nucletide identities of 99.6%-99.8% while the sixth stains closest with strains isolated from Myanmar(2002)share the nucletide identities of 98.8%. The isolates have five amino acid mutations compared with strains epidemic in Guangdong province in 2013,three mutations(S88V,E203G,T275R)are in the EⅡdomain and one mutation (S305P)is in the EⅢdomain which associated with virulence.[Conclusions]During the outbreak in Guangdong province in 2014, DENV-1 is the predominant causative serotype,and there are at least two different kinds of genotypes of DENV-1 largely epidemiced in the whole province. Evolution analysis reveals the multiple origins of the isolates which may origin from Guangdong province , Sigapore,Malaysia,Myanmar so that we should enhance the study and surveillance of autochthonous and vectors in order to understand the epidemic way of dengue in Guangdong province. The isolates have had four mutations in the domain associated with virulence which remain further study to know their biological effects.

Objective To investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on voltage-gated calcium channels on the cell membrane of pheochromocytoma cell 12 (PC 12 cells) induced by ischemic/hypoxia and its mechanisms.Methods PC12 cells at logarithmic phase were collected,which were challenged with hypoxia for 6 hours and reoxygenation for 12 hours to stimulate the nerve cells suffered ischemia/reperfusion pathological process in vitro.PC12 cells were divided into non-infection group,infected by lentivirus containing green fluorescent protein (GFP) without HSP70 gene lentivirus control group (GFP lentivirus control group),and infected by lentivirus containing HSP70 and GFP gene recombined lentiviral infection group (HSP70 recombined lentiviral infection group).Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot were used to determine mRNA and protein expressions of L-type voltage-gated Ca2+ channel subunits cav1.2 and cav1.3,receptor gated channel subunits NR1 and NR2,and Na+/Ca2+ exchange (NCX) in PC12 cells.Results After being challenged with hypoxia/reoxygenation,the mRNA and protein expressions of cav1.2,NR1 and NR2 in the PC12 cells were significantly lower in HSP70 recombined lentiviral infection group than those of GFP lentivirus control group and non-infection group [cav1.2 mRNA (2-△△Ct):3.13 ± 0.46 vs.5.12 ± 0.52,5.13 ± 0.66;NR1 mRNA (2-△△△Ct):1.61 ± 0.44 vs.3.23 ±0.82,3.31 ±0.78;NR2 mRNA (2-△△Ct):2.09±0.41 vs.3.91 ±0.64,3.88±0.62;cav1.2 protein (gray value):2.82±0.39 vs.3.98±0.23,3.96±0.24;NR1 protein (gray value):1.84±0.35 vs.2.79±0.21,2.86±0.23;NR2 protein (gray value):0.87±0.24 vs.1.57±0.31,1.33±0.44;all P ＜ 0.01].But there were no statistical differences in the mRNA and protein expressions of cav1.2,NR1 and NR2 between GFP lentivirus control group and non-infection group (all P ＞ 0.01).There were no statistical differences in the mRNA and protein expressions of cav1.3 and NCX among non-infection group,GFP lentivirus control group and HSP70 recombined lentiviral infection group [cav1.3 mRNA (2-△△Ct):4.82 ± 0.32,4.72 ± 0.36,4.82 ± 0.29;NCX mRNA (2-△△Ct):3.49 ± 0.78,3.47 ± 0.71,3.56 ± 0.65;cav 1.3 protein (gray value):2.63±0.40,2.64±0.39,2.68±0.39;NCX protein (gray value):3.27±0.48,3.34±0.48,3.31 ±0.42;all P ＞ 0.01].Conclusion Exogenous HSP70 affects the expression of L-type voltage-gated Ca2+ channel subunit cav1.2 and receptor gated channel subunits NR1 and NR2,which may protect PC12 cells from the injury caused by ischemic/hypoxia.