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 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

The essence of endogenous turbidity in Chinese medicine (CM) is different from cream, fat, phlegm, retention, damp, toxicity, and stasis. Along with the development of modern scientific technologies and biology, researches on the essence of endogenous turbidity should keep pace with the time. Its material bases should be defined and new connotation endowed at the microscopic level. The essence of turbidity lies in abnormal functions of zang-fu organs. Sugar, fat, protein, and other nutrient substances cannot be properly decomposed, but into semi-finished products or intermediate metabolites. They are inactive and cannot participate in normal material syntheses and decomposition. They cannot be transformed to energy metabolism, but also cannot be synthesized as executive functioning of active proteins. If they cannot be degraded by autophagy-lysosome or ubiquitin-prosome into glucose, fatty acids, amino acids, and other basic nutrients to be used again, they will accumulate inside the human body and become endogenous turbidity. Therefore, endogenous turbidity is different from final metabolites such as urea, carbon dioxide, etc., which can transform vital qi. How to improve the function of zang-fu organs, enhance its degradation by autophagy-lysosome or ubiquitin-prosome is of great significance in normal operating of zang-fu organs and preventing the emergence and progress of related diseases.
Autophagy ; Humans ; Medicine, Chinese Traditional ; Proteasome Endopeptidase Complex

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

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

Ductus Arteriosus ; abnormalities ; Fatal Outcome ; Female ; Heart Septal Defects, Atrial ; etiology ; Humans ; Infant, Newborn ; Rubella Syndrome, Congenital ; complications