Animals ; Beer ; adverse effects ; Blood Glucose ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; Lipid Metabolism ; Male ; Physical Conditioning, Animal ; physiology ; Rats ; Rats, Wistar

SMART-RACE was performed after isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase (GenBank Accession No. AJ620046). Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6 ? His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4. 0-8.0 and 20-40℃,wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme, and recombinant expression provided a chance of highly expressing arabinosidase.

Objective To investigate the effects of transplantation of bone marrow mesenchymal stem cells (BMSCs) transfected with adrenomedullin (ADM) on cardiac function in heart failure rats and the mechanism.Methods BMSCs were isolated from femur and tibia marrow of 10 rats,20 days old,body weight 30-50 g,and in vitro cultured.The third passage of BMSCs were tuansfected with adenovirus containing ADM and labeled with green fluorescent protein(GFP).Before transplantation,BMSCs were labeled with 4',6-diamidino-2-phenylindole (DAPI).Eighty healthy male Wistar rats weighted 180-200 g were randomly divided into 2 groups according to body weight:control group (n =10) was injected with normal saline (NS); diffuse myocardial injury heart failure rat model(n =70) was established by subcutaneous injection of isoproterenol (ISO,170 mg/kg) every day for 4 consecutive days.Four weeks after administration of ISO,heart function was assessed by echocardiography,the 39 rats with left ventricle ejection fraction(LVEF) ＜ 70％ of global heart failure model were randomly divided into three groups in accordance with the level of heart function:untransfected group,transfected group and NS group.DAPI labeled untransfected BMSCs suspension,ADM gene transfected BMSC suspensions (3 × 106/150 μl) and equal volume of NS were injected into the left ventricular anterior wall in 4 places in each goup.Control group received thoracotomy only.Four weeks after transplantation,rats were examined by ultrasound echocardiography,then were sacrificed and left ventricular were dissected.The myocardium was stained with Massons trichrome to analyze myocardial tissue fibrosis.The transplanted cells were observed by fluorescence microscopy and matrix metalloproteinase-2 (MMP-2) expression of myocardial tissue was detected in each group by Western blotting.Results After in vitro culture for three days,the BMSCs began to grow adherently,tended to be fused about 10 days,in the fusiform shape.Four weeks after transplantation,ultrasound echocardiography results showed that rat cardiac left ventricular end systolic diameter (LVDs),LVEF,and left ventricular cardiac fractional shortening (LVFS) were different between groups,and the difference were statistically significant(F =5.838,32.983,51.714,P ＜ 0.05 or P ＜ 0.01).Compared with the control group[(86.50 ± 1.54)％,(50.66 ± 1.87)％],the LVEF and LVFS of NS groups[(56.67 ± 6.86)％,(26.27 ± 4.01)％],the transfected group[(79.40 ± 1.70)％,(43.48 ±2.15)％] and untransfected group[(69.24 ± 7.30)％,(34.59 ± 5.13)％] were significantly lower(all P ＜ 0.05);compared with the NS group,the LVEF and LVFS of the transfected groups and the untransfected group were significantly increased(all P ＜ 0.05) ; compared with the untransfected group,the LVEF and LVFS of the transfected group were increased (all P ＜ 0.05).Compared with the control group [(3.16 ± 0.22)mm],the LVDs of the NS group[(5.35 ± 1.57)mm] was significantly increased (P ＜ 0.01); compared with the NS group,the LVDs of the transfected group and the untransfected group[(3.95 ± 0.55),(4.24 ± 0.92)mm] were significantly decreased (all P ＜ 0.05).There was no statistically significant difference between the control group,the transfected group and the untransfected group in LVDs (P ＞ 0.05).It can clearly be seen that there was GFP and DAPI labeled transplanted cells under a fluorescence microscope in the myocardial tissue transplanted area.There was significant difference in myocardial fibrosis area and the myocardial tissue protein expression of MMP-2 between groups(F =533.75,32.777,all P ＜ 0.01).The area ratio of the NS group[(15.200 ± 0.356)％,0.584 ± 0.013],the transfected group[(8.530 ± 0.573)％,0.386 ± 0.017] and the untransfected group [(10.670 ± 0.369)％,0.438 ± 0.015] and the MMP-2 protein expression were significantly higher than that of the control group[(1.070 ± 0.113)％,0.319 ±0.013,all P ＜ 0.01)]; compared with the NS group,the two index of the transfected group and the untransfected group were decreased (all P ＜ 0.05).Compared with the untransfected group,the two index of the transfected group was decreased (all P ＜ 0.05).Conclusion Transplantation of ADM gene transfected BMSCs can improve heart function of rats with heart failure significantly and reduce myocardial fibrosis.

Objective To explore compliance of anti-platelet therapy of patients after coronary stent implantation and relative factors. Methods 75 patients after coronary stent implantation were investigated by APGAR and drug compliance evaluation scale. Results 44.0 ％ of patients (n =33) failed to take anti-platelet drugs exactly following the prescription, the main form is omitting to take anti-platelet drugs (28.0％). The total score of understanding about antiplatelet medication knowledge in 75 patients was (26.36 ± 2.85 ), they took not much knowledge about antiplatelet druys' side effects and adverse consequences. The median adherence score of compliamce of antiplatelet drugs was 15.61. Fully accounted for 56.0％. The main non-complicance behavior was missed 28.0％. Different time of stent placement, antiplatelet druy type, duration of patient understanding of medication and medication compliance in patients with family functioning were statistically significant differences ( P ＜ 0. 05 ) ; Medication compliance and stent placement time was negatively correlated (r =-0. 304,P ＜ 0. 01 ), and duration of antiplatelet drug use to understand the situation, family functioning was positively correlated (r =0. 259,0. 266 ; P ＜ 0. 05 ). Conclusions The rate of good compliance of antiplatelet therapy of patients after stent implantation is much lower. Nurses should emphasize the education about anti-platelet therapy and start family support system in order to improve the compliance of anti-platelet therapy.

Objective To investigate the effects of different standard cross sections and angles on the measurement accuracy of induced postnatal fetal long bones. Methods Fetal long bones (femori and humeri) in 30 cases with induced abortion were measured utilizing ultrasound from different angles and /or at different directions. The values measured from different sections and angles with vernier calipers were compared prenatally and postnatally. Results There was no apparent difference between the pre-induced abortion and those of the post-induced abortion. The results in the 30 cases showed that: (1) the values measured from anterior 90 degree, the long bone length would best match with the bare long bone length up to 96.7%, the match rate of other angles and/or directions was up to 80%; (2) no apparent statistical difference was between the length of left and right bone and no difference was found using 4 different directions and 3 different angles; (3)there was no difference between the left and right femuri and humeri.Conclusions Though the measured value from anterior 90 degree direction was the most accurate one, the statistical analtical results showed no difference among 12 values measured from 3 different angles and/or 4 different directions.

OBJECTIVETo determine the plasma level of Epstein Barr virus (EBV) DNA in children with EBV associated diseases, and to investigate the dynamic changes of EBV DNA level after initial infection as well as the relationship between EBV DNA level and the diseases severity.

METHODSThe subjects consisted of 73 children with primary EBV infection (infectious mononucleosis, pneumonia,etc.) and 18 children with severe EBV-associated diseases (chronic active EBV infection, hemophagocytic lymphohistiocytosis, etc.). The plasma EBV DNA level was detected by a real-time PCR assay.

RESULTSThe plasma EBV DNA level decreased with the infection time in children with primary EBV infection. Two weeks after infection, plasma EBV DNA was almost undetectable. The positive rate of plasma EBV DNA in children with severe EBV associated diseases increased significantly when compared with that in children with primary EBV infection (89% vs 16%; p<0.05).

CONCLUSIONSThe level of EBV replication may be reduced with the infection time. Dynamic determination of blood EBV DNA is useful for the evaluation of disease severity in children with EBV infection.

DNA, Viral ; blood ; Epstein-Barr Virus Infections ; virology ; Humans ; Infectious Mononucleosis ; virology ; Lymphohistiocytosis, Hemophagocytic ; virology ; Virus Replication

China ; Genes, Bacterial ; Sequence Analysis ; Streptococcus suis ; classification ; genetics ; isolation & purification ; Virulence Factors ; genetics

OBJECTIVETo investigate the epidemiological features, genetic drift in the epitopes of hemagglutinin (HA) of the novel influenza A (H1N1) virus and oseltamivir-resistant variants characterized by H275Y and N295S mutations in children in Shanghai since the outbreak.

METHODBetween June 2009 and May 2012, a prospective surveillance study was carried out in Shanghainese children who attended the outpatient clinic of Children's Hospital of Fudan University for influenza-like illness. One-step real-time fluorescence quantitative RT-PCR was performed to detect seasonal influenza A and influenza B virus and the novel influenza A (H1N1) virus in the respiratory samples. Genetic drift from the vaccine strain in HA epitopes of the novel influenza H1N1 virus and the molecular markers associated with oseltamivir resistance in neuraminidase (NA) were analyzed.

RESULTOut of 3475 enrolled cases, the novel influenza A (H1N1) virus was confirmed virologically in 222 (6.4%) otherwise healthy children with 133 (59.9%) being boys and 89 (40.1%) girls. The median ages of children with the novel influenza A (H1N1) virus infection during the first wave from August 2009 to February 2010 and the second wave from December 2010 to February 2011 were 53.5 months and 32.0 months, respectively (Z = -4.601, P = 0.000); 119 (46.9%) had the close contact with persons suffering from fever or respiratory infection, of whom, 68 (57.1%) contacts were family members and 47 (39.5%) contacts were classmates. During the outbreak in 2009-2010 season, 66 (40.9%) were exposed to primary index cases, school students were the major exposure subjects, accounting for 50.0%. The nucleotide sequences of HA1 gene were highly homologous between the vaccine strain A/California/07/2009 and Shanghai circulating novel influenza A (H1N1) strains and only S83P mutation in epitope E of HA was detected inclusively in the circulating strains. The H275Y and N295S amino acid mutations associated with oseltamivir resistance were not found in the circulating novel influenza (H1N1) strains.

CONCLUSIONTwo major waves of the novel influenza A (H1N1) outbreaks occurred in Shanghainese children during 2009-2011. Institutional children were the major affected individuals during the 2009 pandemic wave. Households and schools were the main sites of transmission among children during influenza pandemic. Influenza vaccination should be enhanced in children and their close family contacts. The novel influenza A (H1N1) virus in Shanghai has not undergone significant genetic changes. Oseltamivir is effective for the treatment of the novel influenza A (H1N1) virus.

Adolescent ; Amino Acid Sequence ; Antiviral Agents ; pharmacology ; Child ; Child, Preschool ; China ; epidemiology ; Drug Resistance, Viral ; Female ; Hemagglutinins, Viral ; genetics ; Humans ; Infant ; Influenza A Virus, H1N1 Subtype ; classification ; genetics ; isolation & purification ; Influenza, Human ; drug therapy ; epidemiology ; pathology ; virology ; Male ; Molecular Epidemiology ; Molecular Sequence Data ; Neuraminidase ; genetics ; Oseltamivir ; pharmacology ; Pandemics ; Viral Vaccines ; genetics ; immunology

This study is to investigate the modulation of Kudiezi (KDZ) injection on differential protein expression in cerebral cortex of rats with cerebral ischemic stroke and heat toxin syndrome established by intraperitoneal injection of carrageenan and middle cerebral artery occlusion (MCAO) methods. According to random number table rats were divided into three groups: drug group, model group and sham group. The tripheye tetrazolium chloride (TTC) staining and HE staining were used to observe brain tissue injury of rats. After therapeutic intervention with above drug for seventy-two hours, the level of differential protein expression was analyzed by two-dimensional gel electrophoresis (2-DE). The results show that there are differential protein expressions between cerebral ischemic stroke and heat toxin syndrome rats and sham rats. Furthermore, as a Chinese medicine injection with effect of clearing heat, resolving toxin and dredging collaterals, KDZ injection can decrease alleviate morphological changes of cerebral ischemia, regulate the levels of some differential proteins expression.
Animals ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; pathology ; Cerebral Cortex ; drug effects ; metabolism ; pathology ; Drugs, Chinese Herbal ; administration & dosage ; Gene Expression ; drug effects ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Stroke ; drug therapy ; genetics ; metabolism ; pathology

OBJECTIVETo investigate the effect of AT₁ receptor on the changes of tyrosine hydroxylase-immunoreactivity (TH-IR) in rostral ventrolateral medulla (RVLM) induced by brain cholinergic stimuli in rats.

METHODSMale SD rats were randomly divided into 4 groups: NS + CBC group, Los + CBC group, Los + NS group and NS + NS group. AT₁ was blocked by pretreatment of 20 μg losartan in Los + CBC and Los + NS groups; intracerebroventricular injection of 0.5 μg carbachol was used for cholinergic stimuli in NS + CBC and Los + CBC groups; normal saline (NS) was used for control. The output amount of natrium in kidney, glomerular filtration rate (GFR) and renal plasma flow (PRF) were observed. The changes of TH-IR in the RVLM were observed by immunohistochemistry.

RESULTIn NS + CBC group carbachol induced potent natriuresis, after pretreatment of losartan the natriuretic effect was partially inhibited in Los + CBC group. Both the number and optical density of TH-IR positive neurons in NS + CBC group were markedly increased than those in NS + NS group (P < 0.05); while those in Los + CBC group were significantly lower than those in NS+CBC group (P < 0.05). Intracerebroventricular injection of carbachol and losartan had no effect on GFR and RPF(P > 0.05).

CONCLUSIONThe results suggest that cholinergic stimuli can induce potent natriuresis and increase the activity of adrenergic neurons in the RVLM; the above effects can be down regulated by blockade of brain AT₁ receptor.

Animals ; Carbachol ; administration & dosage ; pharmacology ; Drug Antagonism ; Glomerular Filtration Rate ; drug effects ; Losartan ; pharmacology ; Male ; Medulla Oblongata ; drug effects ; metabolism ; Natriuresis ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptor, Angiotensin, Type 1 ; physiology ; Tyrosine 3-Monooxygenase ; metabolism