Objective To fabricate a novel polyvinyl alcohol (PVA)-chitosan(Cs)-collagen (Col) composite material and to confirm the feasibility of its application as a scaffold in tissue engineering.Methods PVA was blended with chitosan and collagen.The water content,swelling ratio,and tensile strength of the scaffolds were tested.SEM was used to observe the histological modality of the cross section.Results Scaffolds which al'e composite with different molecular weight and different amount of Cs and Collagen ale made,with water content ranging from 60.15% to 72.50% and swelling ratio being from 185.33% to 317.57%.The tensile strength of the composite material is 5.70MPa.The inner modality and structure of the scaffolds varied as the proportion of the chemical components changed.Conclusion PVA-Cs-Col scaffolds have high water content and proper swelling ratio and ale rich in porous structure.When the blending proportion is Cs:PVA:COI=30:15:0.20,the scaffold performs best,which shows to be a suitable structure for tissue engineering scaffold.

AIM: Based on the water-solubility of polyethylene glycol (PEG) with different relative molecular weight (RMW) and sodium alginate, we investigated the porous structure and property of calcium alginate scaffold with different RMW and dosages of PEG, which was used as porogenic agent. METHODS: The experiment was carried out in the Key Laboratory of Biomedical Engineering and Biomaterials, Jinan University from March 2006 to September 2007.①PEG at different RMWs (Mw=2 000, 4 000, 6 000, 8 000, 10 000, 20 000, 35 000) and different dosages (mass fraction=0, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06) were added into sodium alginate solution at 0.02 mass fraction. Then alginate was crosslinked, solidified and molded into indiscerptible film and graininess through Ca2+. PEG was dissolved by water. Therefore, a mass of porous structures could be formed in alginate.②Water content and swelling ratio of alginate was tested. Scanning electron microscope was used to observe the porous configuration. RESULTS: ①Porous alginate scaffold could be obtained through PEG and it exhibited good intensity and toughness, with water content reaching 92%. The scaffold could be formed into film and graininess.②Scanning electron microscope results revealed that the distribution of porous structure was uniformed. The aperture was 43.75 ?m-2.8 mm, and could be controlled by PEG's RMW and dosage. Inflated and uniform aperture structure was harvested when RMW of PEG was 4 000 and 6 000. CONCLUSION: Porous alginate structure can be obtained through regulating PEG's RMW and dosage. High water content and porosity of alginate scaffold material can be used in cell culture of tissue engineering and used as controlled release matrix of bio-active component.

Objective To establish a methamphetamine-dependent model in zebrabfish.Methods On the basis of conditioned place preference (CPP) in drug-dependent experiment,place preference box for zebrafishes was designed.According to the natural characteristics of zebrafishes,their preference side and non-preferred side were determined.After intraperitoneal injection of methamphetamine,zebrafishes were placed in non-preferred side(drug box).After intraperitoneal injection of normal saline,zebrafishes were placed in preference side (non-drug box).CPP training was five days and then the methamphetamine-induced place preference in zebrafish were observed.Results After five days training,the staying time of zebrafishes of control group in drug box was not significantly lengthened to compared with before training(.( 287.5 ± 80.18 ) s,(276.3 ± 85.04) s),P＞ 0.05 ).The staying time of zebrafishes of model group in drug box was markedly extended after training.In comparison with before training or control group,the significant differences were observed ( (465.5 ± 113.49 ) s,( 247.9 ±95.62)s,(276.3 ±85.04)s,P＜0.01).Conclusion Methamphetamine can induce conditioned place preference in zebrafishes.The CPP model in zebrafish established in this study can be used as a new animal model in drug dependence.

Objective To investigate the regulatory networks of DNA methylation profiles in STEMI by methylation microarrays.Methods A total often male patients with STEMI and ten male healthy controls were recruited.Methyl-DNA immunoprecipitation and Nimblegen HG18 Meth 385K promoter plus CpG island microarrays were used to identify differentially methylated regions.And several bioinformatics analysis tools which included chromosomal assignment, gene ontology analysis and pathway analysis with SignalMap and The Database for Annotation, Visualization and Integrated Discovery were used to high-throughput analysis.Results Compared with healthy controls, DMRs of STEMI is 1 634, There are 1 480 (90.57％), 131 (8.02％) and23 (1.41％) methylated sites were separately located on High CpG-containing promoter, Intermediate CpG-containing promoter and Low CpG-containing promoter;Gene Ontology and Pathway analysis expressed DNA methylated genes of signaling pathway in MI identified glycerophespholipid metabolism, cysteine and methionine metabolism, Dilated cardiomyopathy, Arrhythmogenic right ventricular cardiomyopathy, regulation of actin cyteskeleton, calcium signaling pathway.However, the signal pathway about lipid metabolism is shown no significant difference.Conclusions Bioinformatics tools could provide the quick and high-throughput analysis of data from methylation microarray and enable the function classification of differentially expressed genes of STEMI.

ObjectiveTo evaluate the gastroprotective activity of ascaridole.MethodsThe gastroprotective effect of ascaridole was evaluated on ulcer healing in rats with acetic acid-induced chronic gastric ulcer,pylorus ligation- and Aspirininduced gastric ulcer.Ascaridole was ig administered with the dosages of 10 and 20 mg/kg once daily for 7 d.Results Ascaridole showed the significant anti-ulcer effects.In acetic acid-induced gastric ulcer rats,the ulcer areas after 10 and 20 mg/kg of ascaridole treatment were (65.1 ± 20.0) and (50.6 ± 11.0) mm2,respectively,which were significant lower (P ＜ 0.01) than that of the control group [(116.7 ± 35.8) mm2].For pylorus ligation model,ascaridole showed a gastric ulcer healing effect in a dose-dependent manner.Ascaridole at the dose of 20 mg/kg showed 50％ ulcer protection and had a significant (P ＜ 0.05) gastroprotective activity since it decreased the total acidity and pepsin activity.Compared to the control group,the two dosages of ascaridole showed the significant reduction (P ＜ 0.05) in the ulcer index on Aspirin-induced ulcer.ConclusionThis study provides evidence that ascaridole shows potential efficacy on the healing of gastric ulcers induced by acetic acid,Aspirin,and pylorus ligation.

Objective To estimate the effect of the enviromental pollutant 2,3,7,8-tetrachlorodibenzo-pdioxin (TCDD),a representative of the dioxin family,on the expression of cytochrome P4501A1 (CYP1A1) in cultured human immortalized SZ95 sebocytes in vitro,so as to improve understanding of the pathogenesis of chloracne.Methods SZ95 sebocytes were cultured with or without the presence of 10 nmol/L TCDD for two hours or three days.Real time fluorescence-based PCR was performed to quantify the mRNA expression of CYP1A1,immunohistochemistry and Western blot to determine the expression level of CYP1A1 protein,in the SZ95 cells.Chi-square test was done to compare the protein and mRNA expressions of CYP1A1 between untreated and treated SZ95 cells.Results Real time PCR showed that the mRNA expression of CYP1A1 was low in SZ95 sebocytes,and increased by 5.622 times after 2-hour treatment with TCDD(P ＜ 0.05).Immunohistochemistry revealed a weak expression of CYP1A1 protein in the cytoplasm and nuclei of untreated SZ95 sebocytes,which was also significantly enhanced by the TCDD treatment.Western blot results showed that the relative expression level of CYP1A1 protein was 4.233 ± 0.252 in SZ95 sebocytes treated by TCDD for three days,significantly higher than that in untreated sebocytes(0.123 ± 0.208,P ＜ 0.05).Conclusions There is a low expression of CYP1A1 mRNA and protein in SZ95 sebocytes,which can be upregulated by TCDD,suggesting that the CYP1A1 gene is a downstream target of the aryl hydrocarbon receptor responsible for the abnormal differentiation of human sebocytes.

Objectives To investigate clinical features and the risk factors for 30-day death in elderly chest pain patients.Methods In the prospective study,514 patients with acute chest pain leading to emergency department visit were selected from March 2012-August 2010 and grouped into elderly group (aged≥65 years,n=309) and non-elderly group (aged< 65 years,n=205).The patient's clinical data during 30-day follow-up period were recorded for analysis and comparison.Multivariate regression analysis was used to investigate the risk factors of death.Results Among 514 cases with acute chest pain,30(5.8%)patients with all-cause death included 24 cases in group of 309 (7.8%) elderly patients and 6 (2.9%) cases in group of 205 non-elderly patients during 30 day follow-up period.Univariate regression analysis showed that female,low SBP,Killips' classification ≥ Ⅱ,high level of serum troponin T and creatinine,coronary artery ischemia were more likely to died during 30 day follow-up period.And female and Killips' classification ≥ Ⅱwere the independent factor for 30-day death in the elderly[OR:3.55 (95%CI:1.00-12.59) and 5.90 (95%CI:1.31-26.63)]respectively.Conclusions Elderly patients with acute chest pain for first emergency department visit are at high risk for 30-day death.Female and cardiac function Killips' classification ≥ Ⅱ,high levels of serum troponin T and creatinine and coronary artery ischemia are associated with 30-day death in patients with acute chest pain for first emergency visit.Female and Killips' classification ≥ Ⅱare the independent risk factor for 30-day death.

Objective To analyze the changes of lymphocyte subsets in peripheral blood of patients with drug eruption . Methods 18 newly diagnosed patients were served as the drug eruption group ,and were subdivided into cephalosporin group (n=9) ,penicillin group(n=5) and Chinese medicine group(n=4) according to different sensitizing drugs .20 healthy people were taken as the control group .Flow cytometry were utilized to detect the percentages and absolute counts of T lymphocytes (CD3+ ,CD3+CD4+ and CD3+CD8+ ) ,B lymphocytes ,natural killer cell(NK) and natural killer T lymphocytes(NKT) in their peripheral blood . Results Differences of percentages of T lymphocytes (CD3+ ,CD3+ CD4+ ) ,B lymphocytes ,NKT cells between the drug eruption group and the control group showed statistical significant (P<0 .05) .Difference of percentages of CD3+ CD8+ lymphocytes of pa-tients between the drug eruption group and the control group demonstrated no statistical significant (P>0 .05) ,while that of abso-lute counts of T and B lymphocytes of patients was statistical significant between the drug eruption group and the control group (P<0 .05) .Conclusion The percentages of CD3+ ,CD3+CD4+ lymphocytes of patients with drug eruption decrease ,while those of NKT cells increase ,which may be related to the patients′immune regulation .

Objective:To investigate the effect of lipopolysaccharide (LPS) on primary neonatal rat cardiomyocytes (CMs) and human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs).Methods:The hiPS-CMs and primary neonatal rat CMs were treated with different concentrations of LPS for 24 to 48 h. Then the cellular viability was analyzed by the xCELLigence RTCA Cardio system. The measurement of NPPB gene was studied by qRT-PCR and the gene expression analysis was performed by the qPCR array, in order to evaluate the cardiac inflammation effect induced by LPS.Results:The LPS exposure led to dysfunction in the primary neonatal rat CMs, which shown as an increase in beating rate and a decrease in contraction amplitude ( P<0.01), accompanied by an increased NPPB mRNA level ( P<0.01). There was no significant alteration in beating rate and the contraction amplitude in the corresponding concentration of the primary neonatal rat CMs ( P>0.05), as well as the NPPB mRNA level ( P>0.05). However, the expression of NPPB mRNA in hiPS-CMs was significantly different at a higher concentration of LPS (5 μg/mL~40 μg/mL) ( P<0.01), but the beating rate and the contraction amplitude showed no significant change, even the concentration of LPS up to 40 μg/mL ( P>0.05). Finally, the genes of C3, Gpnmb, Atf3, Il6r and Ly96 upregulated to 1.5 folds in the primary neonatal rat CMs. In comparison with primary neonatal rat CMs, the AK4, TOLLIP, SPP1, FABP1, IL6R, LY96 and C3 were over expression to 1.5 folds in the hiPS-CMs. Conclusions:In comparison with primary neonatal rat CMs, hiPS-CMs are markedly less injured by LPS and show a different pattern of inflammation gene expression.

Objective To investigate the characteristics of liver enzyme between acute graft-versushost disease (aGVHD) and non-aGVHD groups after allogeneic hematopoietie stem cell transplantation (allo-HSCT). Methods The liver enzyme data of patients with or without aGVHD was analyzed. Results Among the 371 patients, 158 developed aGVHD(41.6%) with a median time of 29. 5 d. In non-aGVHD group, the median elevating times of ALT, AST, GGT, LDH were all ≤ 20 d after transplantation, which were significantly earlier than those of aGVHD group. The peak value of AST was much higher in aGVHD group, but the remaining liver enzyme showed no significant difference. In aGVHD group, the duration of AST, GGT and LDH was significantly longer than the non-occurrence of aGVHD group, but ALT and ALP showed no difference in duration. In ALT, AST, ALP and LDH elevating group, the occurrence rate of aGVHD was significantly higher, but only ALT and LDH elevation could enter logistic regression model. The sensitivity and veracity of ALT or LDH elevating only were not very good for the diagnosis of aGVHD, but if ALT and LDH beth elevated after day 24 and persisted more than 22 d, the sensitivity and veracity were better. Conclusions The change of liver enzyme is common in allo-HSCT and associates with the occurrence of aGVHD, and the liver enzyme elevating only may not be a good diagnostic index of aGVHD. If the dynamic characteristics of liver enzyme is taken into account and the effect of drug-induced liver injury could be excluded, the elevation of liver enzyme still have the diagnostic value of aGVHD.