Background Cardiomyocytes apoptosis was related to oxidative stress which has been shown to play an important role in ventricular remodeling after myocardial infarction(MI).Probucol is a hypolipidemic and antioxidant agents.Several reports demonstrated its cardioprotective effect after MI.However,the exact effect of probucol on the modification of the apoptosis related gene Bcl-2 and Bax is not clear.Objective To investigate the effects of probucol on mRNA and protein expression of Bcl-2,Bax and oxidative stress in myocardial infarcted rats.Methods Forty-one SD rats that survived 24 h after ligating left anterior descending coronary artery were randomly to receive placebo-saline(5 mL/d,n=20)or probucol(probucol 60 mg/kg?d,n=21).Twelve rats underwent sham operation were served as control(n=12).Six weeks after treatment,hemodynamic pararmeters and left ventricular function were measured with catheterization.Cardiomyocytes apoptosis were determined by TUNEL method.Myocardium mRNA of Bcl-2 and Bax expression levels in the non-infarcted myocardium were as- sessed by RT-PCR.The myocardium protein expression of Bcl-2 and Bax in the non-infarcted myocardium were de- termined by Western blot.Colorimetry was used to determine oxidative metabolism index in myocardium.Results 1)Compared with the sham rats,all MI rats showed marked decrease in Bcl-2 mRNA and protein expression in myo- cardium with increase of Bax mRNA and protein expression and apoptosis index(P

AIM: To analyze the effect of human amniotic homogenate extract on corneal neovascularization after corneal alkali burn in the process of pigment epithelium derived factor (PEDF) and vascular endothelial growth factor (VEGF) expression and the effect of corneal neovascularization.METHODS: Totally 32 patients with corneal alkali burn were selected from June 2015 to June 2016 in Foshan,and were randomly divided into Group A and Group B,with a total of 37 eyes.Group A of 17 cases,with a total of 19 eyes,were treated with 40mg/L human amniotic homogenate extract;Group B (n=15),and 18 eyes,treated with 3g/L prednisolone eye drops.In the treatment of 1,4,7,14,21 and 28d at different time points,we observed the growth of corneal neovascularization,and detected the expression of PEDF and VEGF during angiogenesis.RESULTS: Group A of patients in the use of human amniotic homogenate extract after the treatment,the expression level of PEDF was significantly higher than that in Group B(P=0.001),after 28d treatment,the expression level of PEDF reached 0.721±0.314.While patients in Group B the expression level of PEDF was only 0.538±0.253.Two groups had significant difference between the expression level of PEDF (P<0.05).The expression level of VEGF in Group A was lower than in Group B at different time points in the test.After the treatment of 28d patients in the Group A,the expression level of VEGF was 0.152±0.020,in Group B the expression level of VEGF was0.302±0.031.Two groups of patients with VEGF expression level between the differences were statistically significant (P<0.05).The patients number in Group A with corneal neovascularization was significantly lower than that in Group B,the difference was statistically significant (P<0.05).CONCLUSION: Human amniotic homogenate extract can increase the expression of PEDF in corneal neovascularization after corneal alkali burn,inhibit the expression of VEGF and the proliferation of corneal neovascularization.

Objective To investigate the dynamic alterations of parameters of platelet in newborns with pulmonary hemorrhage(PHN).Methods One hundred and forty-eight cases were selected into research group,within 48 hours after admission,every 3 hours at the time of pulmonary bleeding,peripheral arterial blood were collected and tint deal blood samples were examined with full-automatic blood cell analyzer in order to monitor dynamically changes of platelet and its parameters.Results The blood platelet count(BPC),thronbocytocrit and PDW were greatly changed at 6 hours before occuring pulmonary hemorrhage(all P

To improve ethanol production in Saccharomyces cerevisiae,an integration plasmid pUPGKAT with PGK promoter(phosphoglycerate kinase promoter),adh1 gene(the coding sequences of alcohol dehydrogenaseⅠ) and CYC1 terminator(Cytochrome c transcription terminator) was constructed.Firstly,a fusion fragment composed of PGK promoter and adh1 gene was generated by over lap extension PCR and ligated into pUG6 resulting in plasmid pUPGKA.Subsequently,CYC1 termi nator was amplified from pSH65 by PCR and ligated to the SpeⅠand SacⅡrestriction site of pUPGKA.To integrate PGK-adh1-CYC1 into S.cerevisiae genome,pUPGKAT was digested by TthⅢⅠand the lin-earized plasmid was used to transform S.cerevisiae YS2-△adh2(adh2 disrupted strain) by lithium acetate method.The yeast mutant YS2-△adh2-adh1 which had the adh1 gene placed under the PGK promoter and harbored the adh2 deletion was constructed.Anaerobic fermentation showed overexpression of adh1 by PGK promoter resulted in a 8.84% higher ethanol production compared to YS2-△adh2.

Acyl carrier protein is an essential component involved in the biosynthesis of DHA(Docosahexaenoic Acid) via PKS(Polyketide synthase) pathway,which takes the growing acyl chain from one enzyme to another.One cDNA clone,with high homology of ACP,was isolated from Schizochytrium sp.FJU-512 cDNA library.The deduced amino acid sequence contained 142 residues with isoelectric point of 5.04 and had the 4'-phosphopantetheine prosthetic(4'-PP) binding site.The target fragment was digested with BamHⅠ/HindⅢand inserted into the expression vector pET-30a resulting in the plasmid pET-30a/acp.The recombinant vector was transformed into E.coli BL21(DE3) and induced by IPTG.SDS-PAGE analysis demonstrated that ACP was effectively expressed.

Objective To evaluate the pattern of energy metabolism and nutrients intake in patients with chronic viral hepatitis and posthepatitic cirrhosis to effectively direct their nutrition therapy.Methods Resting energy expenditure (REE) was measured with open-circuit indirect Jorimetry in 60 patients with chronic viral hepatitis and 60 patients with posthepatitic cirrhosis.Their normal basal energy expenditure (BEE) was predicted by Harris-Benedict equation and energy intake (EI) was determined by diet recall. Correlation between REE and indicators for nutrition assessment was analyzed.Results REE was (77? 21) kJ?kg~(-1)?d~(-1) in 60 patients with pusthepatitic cirrhosis,significantly lower than BEE[(95?16) kJ? kg~(-1)?d~(-1)(P0.05,and their EI was (127?34) kJ?kg~(-1)?d~(-1),1.41?0.43 times as REE,in which PROI was (1.02?0.29) g?kg~(-1)?d~(-1),1.31?0.61 times as PROE (0.87?0.34) g?kg~(-1)?d~(-1),also indicating a negative nitrogen balance (-2.02?4.07).REE,EI and intake of three nutrients,serum level of albumin and prealbumin (PA) and body weight significantly decreased in patients with posthepatitic cirrhosis,as compared to those in patients with chronic viral hepatitis (P

OBJECTIVETo observe the effect of Fn-TPO gene modification on human bone marrow mesenchymal stem cells (MSCs).

METHODSRetroviral vector containing Fn-TPO gene was constructed and bone marrow MSCs was modified by this vector. The transcription of Fn-TPO gene in MSCs was observed. The proliferation capacities, hematopoietic cells adhering capacities and TPO secretion capacities of gene modified MSCs were assayed respectively. Cord blood CD34 cells were seeded on the gene modified MSCs layers and several essential growth factors were added. After co-culturing in vitro for 7 days, the number of CD34 cells and their colony forming capacities were assayed by flow cytometry and semisolid culture assay.

RESULTSRetroviral vector containing Fn-TPO gene was successfully constructed and bone marrow MSCs were modified by this vector. Fn-TPO gene was expressed by bone marrow MSCs after gene modification. The viability of MSCs had no significant difference between pre- and post-gene-modification [(7.18 +/- 0.89) 10(4)/ml vs. (6.92 +/- 0.77) 10(4)/ml, P > 0.05]. The hematopoietic cells adhering ability of gene modified bone marrow MSCs was reinforced(0. 188 +/- 0.018 vs. 0.167 +/- 0.017, P < 0.01). The concentration of TPO in the MSCs culture supernatant raised from (5.58 +/- 0.37) ng/ml to (7.46 +/- 0.59) ng/ml (P < 0.01) and did not significantly decline in a short-time period, but influenced by the growth status of MSCs. After co-culturing with gene modified MSCs for 7 days, the absolute number of nucleated cells, the percentage of CD34+ cells and the colony numbers of BFU-E, CFU-GM, CFU-GEMM were (29.9 +/- 2.7) x 10(4), (33.3 +/- 2.8)% , 109.3 +/- 4.1, 163.7 +/- 7.1, 13.3 +/- 1.5, respectively, being significantly higher than that co-cultured with non-modified MSCs.

CONCLUSIONSFn-TPO gene modification can improve the capacity of human bone marrow MSCs for hematopoietic cells adhering, TPO secretion and cord blood CD34 cells amplification.

Bone Marrow Cells ; cytology ; metabolism ; Cell Adhesion ; Cell Proliferation ; Cells, Cultured ; Fibronectins ; genetics ; Gene Fusion ; Genetic Vectors ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Retroviridae ; genetics ; Thrombopoietin ; genetics ; metabolism ; Transfection

OBJECTIVETo explore the effect of Se-riched soybean peptide (SSP) on antioxidant function in rats of fatty liver caused by high-fat diet.

METHODSForty Wistar rats were divided into 4 groups randomly and fed with standard diet and water (NC), high-fat diet and water (HC), high-fat diet and SSP (0.1 g/d) (SeH), standard diet and SSP (0.1 g/d) (SeN) respectively. After 10 weeks, the rats were killed to investigate the pimelosis level in liver tissues by Sudan III staining and the expression of hepatic GRP78 by immunohistochemical analysis. We also analyzed the changes of liver function, blood lipid, the glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in livers and serum.

RESULTSThe pimelosis level, total cholesterol (TC), triglyceride (TG), MDA contents and the expression of GRP78 in HC group were significantly higher than those in NC, SeN, SeH groups. The activities of GSH-Px and SOD in liver and serum were markedly up-regulated in SeH (P < 0.01). There was no significant difference between NC and SeN groups.

CONCLUSIONSSP can improve liver cell injury and the antioxidant functions in rats with fatty liver effectively and decrease the expression of GRP78 in liver.

Animals ; Antioxidants ; metabolism ; Diet, High-Fat ; adverse effects ; Disease Models, Animal ; Fatty Liver ; chemically induced ; metabolism ; Heat-Shock Proteins ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Rats ; Rats, Wistar ; Selenium ; pharmacology ; Soybean Proteins ; pharmacology ; Soybeans ; chemistry

Neuritin is a new neurotrophic factor found recently. In order to identify the function of Neuritin clearly, the coding sequence of human neuritin was amplified by PCR from neuritin cDNA , this fragment digested by NocI and NotI was inserted into pET32a by T4 ligase and transformed into E. coli BL21 then the recombinant plasmid named pET32a-neuritin was constructed successfully . Neuritin was expressed distinctly after inducing by EPTG. The product was identified as neuritin by SDS-PAGE and Western blot analysis . The expression production was purified on Ni2+-NTA column.

Objective To investigate the proportion and function of CD_4~+ CD_(25)~+ regulatory T cells (CD_4~+ CD_(25)~+ Tr)in unexplained recurrent spontaneous abortion(URSA).Methods(1)Proportion measurement:the proportion of CD_4~+ CD_(25)~+ Tr cells in peripheral blood was measured by double-label flow cytometric analysis.The samples were taken from 15 URSA women,15 normal non-pregnancy women and 13 normal pregnancy women.(2)Function measurement:CD_4~+ CD_(25)~+ Tr ceils and CD_4~+ CD_(25)~+ T ce]ls were extracted from peripheral blood lymphocytes by the microbeads separation.The purity of CD_4~+ CD_(25)~+ Tr cells and CD_4~+ CD_(25)~+ T cells was measured by flow cytometry.The growth inhibitory effect of CD_4~+ CD_(25)~+ Tr cells on CD_4~+ CD_(25)~+ T cells was assessed in vitro.Results The proportion of CD_4~+ CD_(25)~+ Tr cells was decreased significantly in URSA women(6.9?1.8)% than that in normal non-pregnancy women[(10.8?1.1)%] (P0.05).Conclusion The results suggest that decrease in proportion and function of CD_4~+ CD_(25)~+ Tr cells may be associated with URSA.