Objectives:The present study in vestigated the presence and significance of serum antibody against human myocardial mitochondria in patients with virus myocarditis (VMC) and dilated cardiomyopathy (DCM). 　　Methods:The human myocardial mitochondtia were used as antigen to detect serum autoantibody by immuno-dot blot in 29 VMC patients,24 DCM patients,33 patients with other cardiac (OCD) and 20 healthy blood donor (HBD).The antigen molecular weight was detected by Western blot. 　　Results:①Positive autoantibody against human myocardial mitochordria was found in 41.4% in VMC and 41.7% in DCM patients,which were much higher than 6.1% in OCD or 0% in HBD patients.②Cardiac troponin T was elevated in 43.8% of autoantibody-positive VMC and DCM patients,which was much higher than 12.0% in autoantibody-negative VMC and DCM patients (p＜0.05).③The antigen molecular weight of human myocardial mitochondria was 30KD. 　　Conclusions:①The presence of autoantibody against supports that concept that is responsible for the development of VMC and DCM.Autoantibody is one of the factors that give rise to cardiac injury.The antibody detection may serve as a diagnostic index for VMC and DCM.②The specific antigen is probably human myocardial adenine nucleotide translocator.

Objective:To evaluate the efficacy and safety of selective intra-arterial thrombolysis with urokirmse for acute cerebral infarction.Methods:Thirty-eight patients with acute cerebral infarction were treated by selective intra-arterial thrombolysis with urokinase within 6 hours of onset.Head CT seans were performed immediately and 24 hours after the procedures to find out whether intracerebral hemorrhage(ICH)had occurred.The activities of daily living and functional outcome of the patients were evaluated by the Barthel Index(BI)and the National Institutes of Health Stroke Scale(NmSS).Results:Of the 38 patients,21 achieved complete recanalization,8 achieved partial recanalization,3 occurred symptomatic intracranial hemorrhage,and l died.At day 90,22 patients had a favorable outcome(BI≥90),11(50≤BI＜90)had a better outcome,and 4 had a worse one(BI＜50);at 6 months,24 patients had a favorable outcome,9 had a better outcome.and 4 had a worse one.The NIHSS scores at day 14 after the procedures were significantly superior to the levels betore the procedures(12.68±7.43 versus 15.38±4.32,P＜0.011.Conclusions: Intra-arterial thrombolysis with urokinase can recanalize the occluded vessels and improve the acute clinical symptoms and long-term prognosis of patients.

OBJECTIVE: To investigate the effects of arsenic trioxide (ATO) on the expressions of vascular endothelial growth factor (VEGF) and P-glycoprotein (P-gp) in K562/A02 cells and to explore the correlation between VEGF and P-gp. METHODS: The inhibition rate of K562/A02 cell proliferation was detected by using methyl thiazolyl tetrazolium assay (MTT); the level of VEGF was detected by enzyme-linked immunosorbent assay (ELISA) and the expression rate of P-gp was determined by flow cytometry (FCM). RESULTS: 0.05 micromol/L ATO had no influences on the cell proliferation and the expression of VEGF in K562/A02 cells; 0.4 and 3.2 micromol/L ATO could significantly inhibit the K562/A02 cell proliferation and down-regulate the expression of VEGF in K562/A02 cells (P<0.05). The expression of P-gp did not changed after being exposed to 0.05 and 0.4 micromol/L ATO for 24, 48 and 72 hours (P>0.05). 3.2 micromol/L ATO could remarkably reduce the expression of P-gp in K562/A02 cells after 48- and 72-hour incubation with ATO (P<0.05). CONCLUSIONS: The down-regulation of P-gp and VEGF after being exposed to ATO probably contributes to the reversion of multidrug resistance in K562/A02 cells. The level of VEGF may be related to the expression rate of P-gp in K562/A02 cells.

BACKGROUND:Exogenous neurotrophic factors or chemical induction can induce rat bone marrow mesenchymal stem cells to differentiate into neuron-like cells. However, exogenous inductors exert a short inducible action, and their chemical substances inevitably have a negative impact on cellviability to limit the application prospects of bone marrow mesenchymal stem cells to a certain extent.
OBJECTIVE:To investigate the effect of glial cellline-derived neurotrophic factor, green fluorescent protein gene transfection by adenovirus vector on biological characteristics of rat bone marrow mesenchymal stem cells, to observe the expression of glial cellline-derived neurotrophic factor and green fluorescent protein and the role of nutrition on bone marrow mesenchymal stem cells, and to explore the ability to differentiate into neuron-like cells induced by glial cellline-derived neurotrophic factor.
METHODS:The bone marrow mesenchymal stem cells at passage 3 were transfected by recombinant adenovirus (Multiplicity of infection=10, 50, 80, 100, 150, 200). The experiment had two groups according to target genes:bone marrow mesenchymal stem cells were transfected by Ad-GDNF-GFP in transfection group, and bone marrow mesenchymal stem cells were not transfected in control group. The expression of green fluorescent protein was detected by inverted fluorescence microscope. Transfection efficiency was calculated by flow cytometry. cells viability and the morphological changes of cells were compared respectively by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and inverted fluorescence microscope between the two groups. On days 5 and 10 after transfection, the expression of glial cel-derived neurotrophic factor mRNA was detected by PCR. On day 5, the expression of neuron-specific enolase was determined by immunofluorescence examination. On day 10, the expression of microtubule-associated protein 2 was identified.
RESULTS AND CONCLUSION:By the end of 12 hours after transfection, the green fluorescent protein expressed in cells, and the fluorescence intensity gradual y increased with time. When the multiplicity of infection was 100, the fluorescence intensity was strong and stable, and the transfection rate was nealy 90%on day 3 after transfection. cellviability in the transfection group was strengthened after transfection. On day 5 after transfection, bone marrow mesenchymal stem cells expressed neuron-specific enolase, and neuron-like protrusions gradual y extended. On day 10 after transfection, bone marrow mesenchymal stem cells expressed microtubule-associated protein 2 and glial cellline-derived neurotrophic factor mRNA, and exhibited neuron-like morphology and interconnected synpases. The recombinant adenovirus, Ad-GDNF-GFP, can highly transfect bone marrow mesenchymal stem cells when the multiplicity of infection is 100, and glial cellline-derived neurotrophic factor can promote the proliferation of bone marrow mesenchymal stem cells and induce bone marrow mesenchymal stem cells to differentiate into neuron-like cells.

This study performed phylogenic analysis on a dengue strain isolated from an outbreak of dengue fever in 2009 at Yiwu City of Zhejiang Province,China,and further to analyze the immunogenicity of E protein of this viral isolate.Firstly,the viral genome was amplified by RT-PCR and phylogenetic trees were constructed by MEGA 4 based on both nucleotide and amino acid sequences of E and NS1 proteins.The phylogenetic analysis showed that the similarity of Yiwu strain with the Guangzhou GZ1D3 strain and the India GWL-25 strain was over 99％.Secondly,the expression plasmid of E protein was constructed and transfected into 293T cells.The secreted E protein were then purified by sucrose density gradient centrifugation and used to inoculate BALB/c mice.The humoral immunity was evaluated by ELISA and neutralizing antibody analysis.Resuits showed that the E protein of Yiwu strain could induce dengue specific IgG antibodies and neutralizing antibodies.Therefore,the study found that the Yiwu strain was classified into the subtype Ⅲ of dengue virus type 3 (DENV-3),and the E protein of this strain had strong immunogenicity

OBJECTIVETo determine the nature of genomic copy number variations (CNVs) in two fetuses with congenital heart defects (CHD) and explore the correlation between 3q microdeletions and CHD.

METHODSGenomic DNA was extracted from fetal umbilical cord tissue, and chromosome copy number variations were detected by low coverage whole genome sequencing.

RESULTSBoth fetuses had microdeletions of the long arm of chromosome 3. Fetus 1 had ventricular septal defect, cleft lip and palate, and a 1.66 Mb deletion on 3q29. The deleted region encompassed all of the critical genes for 3q29 microdeletion syndrome. Fetus 2 had overriding aorta, ventricular septal defect, and a novel 240 kb deletion on 3q28.

CONCLUSION3q29 microdeletion may result in CHD in combination with cleft lip and palate. Genomic CNVs can be detected by low coverage whole genome sequencing.

Chromosome Deletion ; Chromosomes, Human, Pair 3 ; DNA Copy Number Variations ; Female ; Genetic Testing ; Heart Defects, Congenital ; genetics ; Humans ; Pregnancy ; Prenatal Diagnosis

Objective:To explore the phenotype and mechanism of ferroptosis in steatosis liver ischemia reperfusion injury (IRI) and elucidate the mechanism of targeted inhibition of ferroptosis down-regulating steatosis liver IRI.Methods:First, 20 mice are divided into 2 group according to the different feeding mode: Normal chow diet(NCD group, n=10)and high fat diet(HFD group, n=10). The fatty liver constructs are successful as verified by laboratory tests for relevant indicators and oil red staining.NCD and HFD mice were randomized into two groups sham and IRI.The expression of glutathione peroxidase 4(Gpx4)in liver tissues of each group is detected by Western blot.Morphological phenotypes of mitochondria in liver tissue are observed by transmission electron microscope.Accordingly, it was determined whether ferroptosis occurred in mouse liver IRIs.NCD(n=30)and HFD(n=30)are further randomized into 3 groups: sham(n=10), IRI(n=10)and inhibitor(IRI-fer-1 group, n=10). The serum contents of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)are detected by ALT/AST kits.Hematoxylin-eosin(HE)stain is employed for detecting the degree of liver injury; immunohistochemistry(IHC)and enzyme-linked immunosorbent assay(ELISA)for detecting the levels of inflammatory factors and inflammatory cell infiltration.Similarly, primary hepatocytes from NCD/HFD mice are extracted and the occurrence of ferroptosis is verified by detecting C11-BODIPY(581/591)by fluorescent stain after hypoxia-reoxygenation(H/R).Results:As compared with NCD-IRI group, Gpx4 protein expression declined obviously in HFD-IRI group while serum ALT/AST level spiked markedly( P<0.01). IHC staining of 4-HNE is positive, mitochondrial specific damage is more pronounced and inflammatory infiltration became enhanced.As compared with IRI group, serum level of ALT/AST dropped obviously and infiltration of inflammatory cells and secretion of inflammatory factors are blunted markedly in IRI-fer-1 group( P<0.01). Conclusions:A high degree of ferroptosis and severe inflammatory response during fatty liver IR are features of distinguishing steatosis liver IRI from ordinary liver IRI.Targeted inhibition of ferroptosis lowers inflammation and damage during IR in steatosis liver.

Objective To investigate the prevalence of dyslipidemia in subjects with type 2 diabetes mellitus in Beijing urban communities.Methods Total 3316 subjects with type 2 diabetes (age 20-80 years) were recruited from 15 urban community health centers in Beijing using a multi-stage random sampling approach.Dyslipidemia was diagnosed according to Chinese Guidelines on Prevention and Treatment of Dyslipidemia in Adults:2007 version.Results Among 3316 diabetic subjects (1329 malesand 1987 females),75.6％ (2506/3316) had dyslipidemia,the prevalence was 72.5％ (964/1329)in men and 77.6％ (1542/1987) in women.The prevalence of hypertriglyceridemia and hypercholesterolemia was 41.9％ (1388/3316) and 48.1％ (1595/3316),respectively.31.5％ (1043/3316) subjects had high levels of low-density lipoprotein cholesterol (LDL-C) and 21.2％ (703/3316) had low high-density lipoprotein cholesterol (HDL-C).Among all subjects with dyslipidemia only 22.9％ (575/2506) took hypolipid agents.The overall blood lipid control rates of triglyceride (TG),total cholesterol (TC),LDL-C and HDL-C in 1393 subjects with dyslipidemia history were 48.0％ (669/1393),17.4％ (242/1393),30.9％ (430/1393) and 75.8％ (1056/1393),respectively.Diabetics with dyslipidemia had higher body mass index,waist circumference,blood pressure,plasma glucose and hemoglobin A1c.The prevalence of dyslipidemia in the overweight and uncontrolled-glucose group were 79.0％ (1678/2125),78.9％ (1756/2227),respectively.Logistic regression analysis showed that gender,age,body mass index and hemoglobin A1c were associated with dyslipidemia.Conclusions The prevalence of dyslipidemia in diabetic subjects in Beijing urban communities is high and less than one quarter patients take hypolipid agents.Age,body mass index and hemoglobin A1c are the risk factors of dyslipidemia in type 2 diabetic patients.

Objective To investigate current status of use of oral hypoglycemic drugs and insulin among adult patients with type 2 diabetes mellitus (T2DM) in urban community of Beijing.Methods In total,3297 T2DM patients aged more than 20 years from 15 urban communities of Beijing were studied.Their body weight,height,fasting plasma glucose level and glycosylated hemoglobin Alc (HbAlc) were measured.A door-to-door questionnaire survey on use of oral hypoglycemic drugs and insulin was conducted for them.All the T2DM patients surveyed were divided into four groups based on their received intervention.Results ①Of 3279 T2DM patients,454 (13.8％) received lifestyle intervention,971 (29.5％) used only one oral hypoglycemic drug,1179 (35.7％) with combined oral hypoglycemic drugs,and 693(21.0％) with insulin.②There was significant difference in average HbAlc among the four groups of T2DM patients with lifestyle intervention,only one oral hypoglycemic drug,combined oral hypoglycemic drugs,and insulin,with HbAI c of (7.0 ± 1.9) ％,(7.1 ± 1.5) ％,(7.4 ± 1.5 ) ％,and (7.5 ± 1.5 ) ％for them,respectively ( F =15.1,P ＜ 0.01 ).Proportions of the T2DM patients with HbAlc equal to or higher than 7.0％ were 32.2％,39.4％,52.1％ and 59.5％ for the four groups,respectively ( x2 =117.7,P ＜ 0.01 ).③In the T2DM patients with lifestyle intervention,32.2％ (146/454) of them with HbA1 c equal to or higher than 7.0％ were untreated with any oral hypoglycemic drug.In those with only one oral hypoglycemic drug,39.4％ (383/971) of them with HbAlc equal to or higher than 7.0％ were not treated with combined oral hypoglycemic drugs and/or insulin.In those with combined oral hypoglycemic drugs,52.1％ (614/1079) of them with HbAlc equal to or higher than 7.0％ were not received combined insulin treatment.④ Fasting plasma glucose level,treatment strategies,postprandial 2-h blood glucose level and length of the illness were independent risk factors for HbAlc level equal to or higher than 7.0％,with odds ratio (OR) of 1.757,1.256,1.175 and 1.031,respectively.⑤ In 2843 T2DM patients with oral hypoglycemie drugs and/or insulin treatment,1494 (52.6％ ) received biguanides and 693 received (24.4％ )insulin,respectively.Conclusions More than half of adult patients with T2DM do not meet the target of glycemic control of HbAlc less than 7.0％ in urban communities of Beijing,due to not active use of oral hypoglycemic drugs,and not timely adoption of combined use of oral hypoglycemic drugs and insulin therapy.

Objective To study clinical characteristics of type 2 diabetic(T2D)patients with metabolic syndrome(MS)and its components in Beijing urban communities.Methods Totally,3295 T2D patients involved in a combined prospective diabetic management study from 15 urban communities in Beijing were classified as four groups, according to 2004 Chinese Diabetes Society's definition of MS, i. e, isolated T2D, T2D with one component of MS, T2D with two components of MS and T2D with three components of MS. Their clinical characteristics were analyzed. Results ( 1 ) Among 3295 T2D patients, 155 (4. 7% )were isolated T2D, 107 (32.6%) T2D with one component of MS, 1386 (42.1%) T2D with two components of MS and 679 (20.6%) T2D with three components of MS, with an overall 62.7% (2065/3295) of T2D patients complicated with MS. (2) In these T2D patients, the more components of MS they had, the higher body mass index (BMI), waist circumference, waist to hip circumference ratio (WHR),systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting serum levels of insulin and triglyceride (TG) and the lower level of high-density lipoprotein-cholesterol (HDL) were presented (P <0. 01 ). (3) Percentage of isolated T2D in women increased from 49. 0% (76/155) to 61.9% (420/679)of those with three components of MS ( P < 0 01 ), with increasing of components of MS. (4) Multiple logistic regression analysis showed that BMI, history of hypertension, decreased HDL, increased TG,increased blood pressure, all were risk factors for T2D patients complicated with MS. Conclusions Among T2D patients in urban communities of Beijing, 95.3% (3140/3295) of them complicated with one or more components of MS, and 61.9% (420/679) of them complicated with MS. So, community diabetic management must be implemented in an all-round way, including control of blood pressure, blood lipids,body weight and so on, in addition to control of blood sugar.