Objective To observe the poisoning components in plasma and histological changes of rabbits with acute toxicity of aconitum kusnezoffii (草乌).Methods Eight rabbits were garaged with aconitum kusnezoffii liquor,aconitum poisoning model was reproduced,electrocardiogram (ECG) and blood pressure were recorded,the concentrations of aconitine,hypaconitine and mesaconitine in plasma after 0.5,1, 2,3 and 6 hours were measured,and the pathological changes of heart,liver and cerebral cortex were observed.Results After garage with poisoning liquor,arrhythmias and the declination of blood pressure, presenting a tendency of progressive aggravation [before garage:(121.98?16.77)/(110.66?8.78) mm Hg, 1 hour after garage:(102.98?8.34)/(90.22?5.85) mm Hg,2 hours after garage:(66.81?9.13)/ (53.40?6.32) mm Hg,1 mm Hg=0.133 kPa,all P

Objective To investigate the changes of serum cardiac troponin I(cTnI)and brain natriuretic peptide(BNP)in heart failure of children with pneumonia and their relationship with heart function.Methods Thirty healthy children aged from 5 months to 3 years old were randomly selected with 17 male and 13 female(healthy group).Thirty children with severe heart failure aged from 3 months to 2 years old were selected at the same time with 21 male and 9 female(heart failure group).Thirty children with ordinary pneumonia aged from 3 months to 3 years old were also sampled with 16 male and 14 female(ordinary pneumonia group).The peripheral bloods of 2-3 mL of all children were taken.The BNP level were measured by enzyme-linked immunosorbent assay and the cTnI level was determined by micro-particle enzyme immunoluminescent.Left ventricular ejection fraction(LVEF) and left ventricular fractional shor-tening(LVFS)were detected by echocardiography.SPSS 11.0 software was used to analyze the data.Results The levels of cTnI [(0.389?0.030) ?g/L] and BNP [(0.572?0.090) ?g/L] of heart failure group increased significantly compared with healthy and ordinary pneumonia group,while their LVEF and LVFS decreased significantly(Pa

AIM: To observe the efficacy of surgical excision combined with autologous limbus conjunctival flap transplantation in the treatment of pterygium accompanied with conjunctival cyst.METHODS: Totally 126 patients 188 eyes with pterygium were hospitalized in Department of Ophthalmology of Tongji Hospital of Huazhong University of Science and Technology during August 2013 and August 2015.The patients were divided into two groups: observation group (11 eyes of 11 patients) with pterygium accompanied with conjunctival cyst and control group (177 eyes of 115 patients) with primary pterygium.All patients underwent slit lamp microscope examination, anterior segment photography, and anterior segment optical coherence tomography(OCT).The size of pterygium was calculated by multiplying neck width and length of the covered corneal.All patients underwent excision combined with autologous conjunctival flap transplantation, and the resections were performed pathological section with hematoxylin and eosin staining.All patients were followed up postoperatively for 4-28mo.RESULTS: All cases in the observation group were confirmed by postoperative pathological examination.All cyst walls were complete, and containing single layer of epithelial cells.The mean size of pterygium of the observation group was 6.9±1.7mm2, and 6.3±1.8mm2 for the control group.There was no significant difference between the two groups (P>0.05).The mean postoperative healing time of observation group was 2.1±0.9d, and 1.9±0.8d for the control group.There was no significant difference between the two groups (P>0.05).Recurrence was seen in two eyes within the follow-up period in the control group, and no recurrence in the observation group.CONCLUSION: Surgical excision combined with autologous limbus conjunctival flap transplantation is a safe and effective treatment for pterygium accompanied with conjunctival cyst.

OBJECTIVETo explore the biological function of vascular endothelial cells from hypertrophic scar, and to analyze the relationship between them.

METHODSThe samples from human hypertrophic scar and normal skin tissue were harvested for histological examination. Then vascular endothelial cells were purified and isolated from the samples, and the level of transforming growth factor (TGF) beta1, platelet derived growth factor (PDGF), endothelin1 (ET)-1, fibroblast growth factor (FGF)2 and vascular endothelial growth factor (VEGF) were determined in a single cell with ELISA.

RESULTSFew capillary vessels were observed in normal skin under microscope, while an increased number of them were present in hypertrophic scar, with slender, tortuous in morphology and even occluded. The diameter of blood capillary in hypertrophic scar was tiny under electron microscope, and the exfoliation of endothelial cells was observed. The levels of TGF-beta1, PDGF, ET-1, bFGF and VEGF from vascular endothelial cells from hypertrophic scar were 60 +/- 8, 30 +/- 4, 0.12 +/- 0.03, 52 +/- 5, 18.1 +/- 1.2 microg/cell, respectively, which were obviously lower than those in normal skin (P < 0.05).

CONCLUSIONThe biological function of vascular endothelial cells was attenuated in the hypertrophic scar, which mightbe the result of the production of large amounts of collagen in the scar tissue, as well as hypoxia.

Adolescent ; Adult ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Collagen ; metabolism ; Endothelial Cells ; metabolism ; Endothelin-1 ; metabolism ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Humans ; Male ; Platelet-Derived Growth Factor ; metabolism ; Skin ; blood supply ; Transforming Growth Factor beta1 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Wound Healing ; Young Adult

Objective To study the expression patterns of Th1 and Th2 cytokine genes in childhood acute idiopathic thrombocytopenic purpura(ITP) and its clinical value.Methods Peripheral blood samples from 30 patients with acute ITP before and after treatment (ITP group) and those from 20 normal healthy subjects (healthy control group) were collected (healthy control group),and reverse transcriptases polymerase chain reaction was performed to determine the mRNA expressions of Th1 and Th2 cytokine genes before and after treatment,which were compared with those of the health controls.Results The positive rate of expression levels of Th1 cytokine genes in samples from ITP patients (3.3％,3.3％) were significantly lower than those from healthy control group (all P ＜ 0.05) and increased to normal level after treatment(16.7％,23.3％).In contrast,Th2 cytokine genes (IL-4,IL-6,IL-10)in the samples from the ITP patients(33.3％,43.3％,40.0％) were significantly higher than those from the healthy control group (all P ＜0.05) and decreased after treatment(10.0％,23.3％,20.0％).Conclusions Such data indicate that ITP is a Th2 cell predominant autoimmune disease,and the abnormal immunity due to Th1/Th2 shift is significant in the pathogenesis of ITP.

CD55 Antigens ; Complement Activation ; Humans ; Membrane Cofactor Protein ; Pemphigoid, Bullous ; Recombinant Fusion Proteins

OBJECTIVE: To investigate the biological characteristics of endothelial progenitor cells (EPCs) from the umbilical cord blood (UCB), and to evaluate their oncogenicity after long-term culture in vitro. METHODS: The mononuclear cells (MNCs) were isolated from the UCB and cultured in MCDB131 medium supplemented with 20% FBS, VEGF and other growth factors. Morphology of the EPCs was observed, and the growth curve of the EPCs was investigated. Surface antigens of the EPCs were analyzed by the flow-cytometer. The capability of intaking the acetylated low-density lipoprotein (acLDL) of the EPCs was detected using fluoresencent chemical method. The vasoformative capability and genetic stability of EPCs were cultured in matrigel, and examined by karyotype analysis. The oncogenicity of EPCs was verified by the tumorigenesis test in athymic mouse and soft agar. RESULTS: EPCs were successfully derived from the UCB, and could be passaged to at least 42(nd) generation and had strong abilities of proliferation, acLDL intake and vasoformation, but there was not oncogenicity. They expressed endothelial cell-surface antigens and maintained normal karyotype. CONCLUSION The EPCs with proliferative potential can be isolated from the UCB. They can be passaged in long-term cultures without oncogenicity, and can maintain normal karyotype. The EPCs can be served as a new type of cells in cell and gene therapy.
Animals ; Antigens, Surface ; analysis ; Cell Line ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Fetal Blood ; cytology ; Flow Cytometry ; HeLa Cells ; Humans ; Infant, Newborn ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Karyotyping ; Mice ; Mice, Nude ; Neoplasms, Experimental ; pathology ; Stem Cells ; cytology ; metabolism ; Vascular Endothelial Growth Factor A ; pharmacology

BACKGROUNDThe interleukin-1 (IL-1) receptor-associated kinase 1 (IRAK1) is believed to play an important role in the pathogenesis of sepsis. Recent studies have suggested that the IRAK1 functional genetic variant could affect the severity of sepsis in Caucasians. In this report, we have investigated whether polymorphisms at the IRAK1 gene are associated with the susceptibility to and severity of sepsis among the Chinese population.

METHODSHaplotype-tagging single nucleotide polymorphisms (htSNPs) were selected from the HapMap database. They were genotyped in 255 patients with sepsis and 260 control subjects by PCR/restriction fragment length polymorphism (RFLP) analysis. The association between the selected htSNPs and the susceptibility to and severity of sepsis were estimated by Logistic regression with adjustments for age, sex, smoking, drinking, chronic disease status, Acute Physiology and Chronic Health Evaluation (APACHE) II score and primary diseases.

RESULTSrs1059702 was selected to represent the six linked htSNPs for IRAK1. Genotype frequencies of the htSNPs were in Hardy-Weinberg equilibrium for females, as were allele frequencies for both sex groups. Associations were observed in females between the htSNPs C/C genotype and increased susceptibility to sepsis (odds ratio (OR), 5.46; 95% confidence interval (CI), 1.12 - 26.67; P = 0.018), and such associations were also observed between the IRAK1 variant haplotype (CC/C-allele) and increased susceptibility to sepsis (OR, 1.68; 95% CI, 1.05 - 2.70; P = 0.031) when compared with the T/T + T/C genotype and the wild-type haplotype (TC + TT/T-allele). In the multiple organ dysfunction syndrome (MODS) subgroup, the variant haplotype was also associated with increased severity of sepsis (OR, 2.37; 95% CI, 1.13 - 4.94; P = 0.02) when compared with the wild haplotype. This association was not significant in male patients.

CONCLUSIONSThe functional polymorphism in exon 5 and the variant haplotype of IRAK1 gene mediate susceptibility to and severity of sepsis. IRAK1 might be a genetic risk factor for the occurrence and development of sepsis in the Chinese population.

Adult ; Asian Continental Ancestry Group ; Exons ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Haplotypes ; genetics ; Humans ; Interleukin-1 Receptor-Associated Kinases ; genetics ; Male ; Middle Aged ; Polymorphism, Restriction Fragment Length ; genetics ; Polymorphism, Single Nucleotide ; genetics ; Sepsis ; genetics

OBJECTIVETo construct a human source vector containing minidystrophin-EGFP fusion gene and investigate its expression in Cos-7 cells.

METHODSThe recombinant human source vector named pHrnDysG was constructed with PCR-clone methods. Three fragments of dystrophin gene were PCR amplified from normal human dystrophin gene cDNA (GenBank NM04006). These three fragments were ligated to generate a minidystrophin gene. The enhanced green fluorescent protein (EGFP) gene was fused to the C terminal of the minidystrophin gene, and then the pHrnDysG was finally obtained by cloning the fusion gene to pHrneo. Fluorescence microscope and RT-PCR were used to detect the expression of minidystrophin-EGFP fusion gene after the recombinant construct was transfected into Cos-7 cells by lipofectamine.

RESULTSRestrictive enzyme digestion analysis and sequencing confirmed that pHrnDysG vector was constructed successfully. After the recombinant pHrnDysG was transfected to Cos-7 cells, RT-PCR demonstrated that the fusion gene was successfully transcribed, and the green fluorescence was observed at the cell membrane.

CONCLUSIONThe minidystrophin-EGFP fusion gene mediated by pHrneo vector could express in Cos-7 cells and its products' localization in the cell membrane was the same as that of full length dystrophin. These results suggested that the recombinant human source vector pHrnDysG might be potentially used in studies on the gene therapy of Duchenne muscular dystrophy.

Animals ; COS Cells ; Cercopithecus aethiops ; Dystrophin ; genetics ; metabolism ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Models, Genetic ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

BACKGROUND: The tumor necrosis factor recepter associated factor (TRAF) 6 is an important intracellular adapter protein that plays a pivotal role in activating multiple inflammatory and immune related processes induced by cytokines. TRAF6 represents a strong candidate susceptibility factor for sepsis. We investigated whether polymorphisms at the TRAF6 gene are associated with the susceptibility to and severity of sepsis. METHODS: A hospital-based case-control study was conducted with 255 patients with sepsis and 260 controls who were recruited from Zhengzhou, China. Haplotype tagging single nucleotide polymorphisms (htSNPs) were selected from the HapMap database and genotyped using the SNPstream genotyping platform. The associations with the susceptibility and disease severity of sepsis were estimated by logistic regression, and adjusted for age, sex, smoking, drinking, chronic diseases status, APACHEII score and critical illness status. RESULTS: A total of 13 TRAF6 SNPs were tagged by 7 htSNPs. Five htSNPs (rs5030490, rs5030411, rs5030416, rs5030445 and rs3740961) were genotyped in the case control study. Genotype frequencies of the htSNPs were conformed to the Hardy-Weinberg equilibrium in both patients and controls. No significant association was found between the 5 htSNPs and the susceptibility to and severity of sepsis. Compared with the main haplotype -11120A/-10688T/-9423A/805G/12967G, no certain haplotype was associated with the significantly susceptibility to or severity of sepsis. CONCLUSION: TRAF6 gene polymorphisms might not play a major role in mediating the susceptibility to and severity of sepsis in the Chinese population. A larger population-based case-control study is warranted.