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 association of MK2 gene with low density lipoprotein cholesterol (LDL-C) and tumor necrosis factor-alpha (TNF-Α) between different gender in Xinjiang Uygur population.

METHODSA total of 350 Uygur males and 595 females were recruited randomly from Hetian area. Two single nucleotide polymorphisms (44890c/t, rs 45514798) in MK2 gene were selected and genotyped by Taqman-PCR in these subjects. All subjects underwent questionnaire-based survey, physical examination, measurement of lipid profiles and plasma TNF-Α determination.

RESULTSAmong the male subjects, the concentration of total cholesterol (TC) [TT vs. CT vs. CC: (4.35±1.20) mmol/L vs. (4.69±1.34) mmol/L vs. (4.83±1.44) mmol/L, P=0.033]and TNF-Α [TT vs.CT vs.CC: (106.63±62.39) ng/dL vs. (128.44±86.15) ng/dL vs. (153.06±82.99) ng/dL, P=0.001]were significantly different in 3 genotypes of 44890c/t. However, the LDL-C levels in TT, CT, and CC genotypes of 44890c/t were not different neither in males nor in females [males: (2.64±1.16) mmol/L vs. (2.81±1.28) mmol/L vs. (3.04±1.32) mmol/L, P>0.05; females: (2.42±1.11) mmol/L vs. (2.36±0.99) mmol/L vs. (2.43±1.05) mmol/L, P>0.05]. None of the allele and genotype frequencies of 44890c/tand rs 45514798 were different between high LDL-C group and control group. Linear regression analysis indicated that body mass index (BMI) (beta=0.089) and TNF-Α (beta=0.092) were significantly associated with LDL-C levels in males (P<0.05), while the age, BMI, and waist/hip ratio with LDL-C levels in females (P<0.05).

CONCLUSIONThe nucleotide polymorphisms (44890c/t and rs 45514798) in MK2 gene may not be associated with LDL-C in both males and females in the Uygur population in Hetian, Xinjiang.

Adult ; Aged ; China ; Cholesterol, LDL ; blood ; Female ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Male ; Middle Aged ; Minority Groups ; statistics & numerical data ; Polymorphism, Single Nucleotide ; Protein-Serine-Threonine Kinases ; genetics ; Tumor Necrosis Factor-alpha ; blood

Sterigmatocystin (ST), the secondary metabolite of many kinds of filamentous fungi, is a potent carcinogen structurally related to the aflatoxins (AFT). With similar chemical structure, sterigmatocystion behaves much the homogeneous properties to aflatoxins, both of these mycotoxins exhibit similar biological properties due to their bisfuranoid structure. Since the common, and even heavier pollution, found in foods and feeds-stuff, sterigmatocystion is more harmful than aflatoxins. The reported detection methods of sterigmatocystion included the Thin-layer Chromatography, the High-Performance-Liquid Chromatography, the Enzyme-Linked Immunosorbant Assay and the PCR detection to the toxic gene, however studies about both easy and inexpensive electro-chemical methods have not been found. Our previous studies had discovered that Sterigmatocystin (ST) exist similar sensitivity towards aflatoxin-detoxifizyme (ADTZ), which we had isolated from a fungus, as aflatoxin does. In this work, the preliminary study on electrochemical analysis and determination of ST with triplet electrode enzyme-biosensor system (Ag/AgCl as the reference electrode, Pt and Au as the pair and work electrode, respectively) was carried out. Multiwall-carbon-nanotube (MWNT) had been used to increase the electron transportation on electrode. In the research, the Au electrode was modified by MWNT-immobilized ADTZ, and then the voltammertric behavior of ST was studied by means of cyclic voltammogram analysis and different pulse analysis. Autoprobe CP Research Atomic Force Microscope and TECNAI 10 Transmission Electron Microscope, had been used to detect the MWNT as well as the surface of MWNT-modified ADTZ. The voltammertric behavior of ST was studied by means of cyclic voltammogram analysis and different pulse analysis. The results show that the red-ox peak potential of ST is at the point of -600 mV, the linear detection range is from 8.32 x 10(-5) to 66.56 x 10(-5) mg/mL, the detection limit is at 8.32 x 10(-5) mg/mL, and the response time is 10 seconds. This study provided a good basic work for further research.
Biosensing Techniques ; methods ; Electrochemistry ; Microscopy, Atomic Force ; Microscopy, Electron, Transmission ; Nanotubes, Carbon ; chemistry ; Sterigmatocystin ; analysis

BACKGROUND: The effect of extracellular matrix on stem cells is the focus of tissue engineering. However, there are few reports about the synthesis and secretion of extracellular matrix as well as its effects on cells. OBJECTIVE: To isolate, culture and identify rabbit bone marrow mesenchymal stem cells (BMSCs), and to explore the changes of extracellular matrix and whole structure under the intervention of ascorbic acid. METHODS: Rabbit BMSCs were isolated by differential adherent method of the bone marrow, and the expression of CD44, CD45 and CD31 was identified by flow cytometry. The BMSCs were cultured in the culture medium containing 20 mg/L ascorbic acid. Then the cell morphology, gross structure, ultrastructure, and histological changes of BMSCs were observed. The expression of extracellular matrix related genes was detected by RT-PCR. RESULTS AND CONCLUSION: Over 95% passage 2 BMSCs could express CD44, but the expression levels of CD45 and CD31 were extremely low. Intervention with ascorbic acid enhanced the proliferation of BMMSCs with unclear cell boundaries. A cell-sheet structure formed at 10-14 days after intervention. Hematoxylin-eosin staining results showed a layered cell arrangement, and Masson staining findings showed a large amount of extracellular matrix composition. Abundant endoplasmic reticula and vesicle-like structure were observed under the transmission electron microscope. RT-PCR findings showed that ascorbic acid significantly increased the expression of fibronectin mRNA in the BMSCs (P < 0.05), but slightly increased the mRNA expression of collagen type I. All these findings indicate that ascorbic acid not only increases the proliferation and transformation of rabbit BMSCs, but also promotes the synthesis and secretion of extracellular matrix, which has great potential in tissue engineering applications.