1.Prediction of Pulmonary Hemorrhage in Newborns by Monitoring Alterations of Parameters of Platelet
min, HU ; yu-jie, ZHANG ; shu-xian, DU
Journal of Applied Clinical Pediatrics 1992;0(06):-
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
5.Effects of Probucol on Expression of Bcl-2,Bax and Oxidative Stress in Myocardial Infarcted Rats
Shu-Xian ZHOU ; Yu-Ling ZHANG ; Yan ZHOU ; Juan LEI ;
Chinese Journal of Hypertension 2006;0(10):-
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
7.Overexpression of Alcohol DehydrogenaseⅠ in Saccharomyces cerevisiae
Li-Na QIN ; Xian-Zhang JIANG ; Bao-Yu TIAN ; Zheng-Yu SHU ; Jian-Zhong HUANG ;
Microbiology 1992;0(02):-
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.
8.Cloning and Expression of Acyl Carrier Protein Gene from Schizochytrium
Zhi-Ping LI ; Xian-Zhang JIANG ; Bao-Yu TIAN ; Zheng-Yu SHU ; Jian-Zhong HUANG ;
Microbiology 1992;0(02):-
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.
9.Effect of fibronectin-thrombopoietin gene modification on human bone marrow mesenchymal stem cells.
Lei ZHANG ; Jie YU ; Shu MO ; Guang YANG ; Xin LI ; Ying XIAN ; Xian-qing JIN
Chinese Journal of Hematology 2007;28(12):832-836
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
10.Effects of gonadotropin-releasing hormone analog treatment on glucose and lipid metabolism in girls with idiopathic central precocious puberty
Long-Jiang ZHANG ; Xiong-Hui XIAN ; Zi-Zhen JIA ; Zhong-Xian BAO ; Shu-Yan WANG
Chinese Journal of Applied Clinical Pediatrics 2013;28(20):1552-1554
Objective To assess the effects of gonadotropin-releasing hormone analog on glucose and lipid metabolism in girls with idiopathic central precocious puberty(ICPP).Methods A total of 26 girls (aged 6-8 years,breast stage B2) with ICPP were followed up in Department of Endocrinology,Shenzhen Children's Hospital from Jan.2008 to Jun.2011.Those girls received triptorelin therapy for 12 months.Before and the end of the 6th month,the 12th month of the treatment,body mass index(BMI) was calculated,fasting plasma glucose(FPG),fasting plasma insulin(FPI),total cholesterol,triacylglycerols,high density lipoprotein cholesterol,low density lipoprotein cholesterol,apolipoprotein A,apolipoprotein B and estradiol(E2) were measured.Insulin sensitivity was estimated by homeostasis model assessment of insulin resistance(HOMA-IR).Twenty age-matched prepuberty girls were set as controls.Results 1.Before treatment,BMI,FPG,FPI and HOMA-IR in ICPP girls had no significant difference compared with the controls.2.After 6 months treatment of triptrolin,serum E2 concentration in ICPP girls declined from(30.5 ± 9.8) ng/L at the beginning of treatment to (11.2 ± 4.6) ng/L at the end of 6th month (P < 0.01) ; The end of 12 th month of the treatment,FPG and FPI had no significant difference compared with that before of the treatment,but BMI increased from(16.46 ± 1.10) kg/m2 to(18.35 ± 1.30) kg/m2,the difference was significant(P <0.05),HOMA-IR increased from 1.24 ±0.30 to 2.08 ±0.40,the difference was significant(P <0.05).3.Lipid metabolism parameters remained unchangeable after 12 months of triptrolin treatment.Conclusion Triptorelin may lead to raise of BMI and HOMA-IR in girls with ICPP at 12 months after treatment.