A simple and effective method for the duplex PCR detection was developed by using sequences of exogenous gene(RDV MP-)and endogenous gene(Zein)as templates for PCR amplification.The results of routine PCR amplification for RDV MP-gene in transgenic maize suggested that RDV MP-gene can stably inheritate in transgenic plants and their progenies;The duplex PCR detection of all negative and part positive samples that obtained by routine PCR amplification confirmed that above negative results were exact,also showed that the quality of extracted DNA can meet the need of PCR amplification.The error ratio of negative samples was 1.4%.The method used in this study was simple and credible and can be used to detect transgenic plants and their products.

Adult ; Herpes Zoster ; etiology ; Herpesvirus 3, Human ; genetics ; Humans ; Male ; Middle Aged ; Mutation

Lymphocyte function associated antigen-1 (LFA-1) is a member of integrin family, that plays an important role in the adhesion of lymphocytes with other cells and matrix. To investigate the role of LFA-1 in collagen-induced arthritis (CIA), the incidence of CIA, histological and radiological assessments in the LFA-1 deficient (LFA-1~ -/- ) mice and control mice were examined. LFA-1~ -/- mice and control mice were immunized with 100?g collagen type II(CII) emulsified with an equal volume of Freund’s complete adjuvant (CFA), followed by the booster injection of the same amount of CII in CFA on day 21. Then, clinical, histological and radiological assessments were done. It showed that 57% control mice developed arthritis and apparently changed in the histological and radiological assessment, whereas the all of LFA-1~ -/- mice had the normal histological and radiographic response and none developed arthritis. These results suggeste that LFA-1 is indispensable for the onset of CIA.

Objective To evaluate the feasibility and efficacy of tumor resection combined with lung volume reduction surgery (LVRS) in pulmonary and esophageal patients.Methods 45 patients with lung tumor and 37 patients with esophageal tumor were di- vided into group A(tumor resection plus LVRS) and group B(tumor resection only) randomly.Group A patients underwent tumor re- section combined with LVRS.The target area was determined according to chest computed tomography and ventilation-perfusion scan- ning.The removed pulmonary tissues made up about 20%～30% of the unilateral lung parenchyma.All patients were followed up for 6～12 months.The pulmonary function,blood gas,dyspnea index and 6-min walk distance were compared.Results There were no death during perioperative period.Forced vital capacity in 1 second,PaO_2,PaCO_2,dyspnea index and 6-min walk distance of the group A were all better than preoperative ones,while group B patients had no statistical difference or slight decrease.Conclusion For some tumor patients with emphysema,simultaneous LVRS might provide better opportunity for surgical treatment,but also im- proved the post-operative quality of life.It widens the operative indication for tumor patients.

Objective To investigate the effects of leukotriene in children with respiratory syncytial (RSV)viral bronchiolitis.Methods The blood and sputum levels of Ieukotriene were detected in 33 cases diagnosed RSV bronchiolitis and 12 cases which were diagnosed pneumonia without RSV infection.Thirty-three cases of bronchiolitis were devided into mild-moderate group(n =22)and severe group(n =11)according to the lowell score.Results The blood and sputum levels of leukotriene in mild-moderate group,severe group,and pneumonia group were(76.96 ± 28.19)pg/ml,(103.53 ± 16.85)pg/ml,(18.14.± 7.49)pg/ml;(31.83 ± 19.14)pg/ml,(67.11 ± 15.11)pg/ml,(6.81 ± 2.90)pg/ml in acute period,and(36.04 ±16.38)pg/ml,(52.27 ± 17.03)pg/m l,(18.14 ± 7.49)pg/ml of serum in recovery period.There were significant differences among three groups(F =48.09,P ＜ 0.001 ; F =15.50,P ＜ 0.001 ; F =44.43,P ＜0.001).After treatment,the blood levels of leukotriene were significantly decreased,but were still higher than that of pneumonia group(P ＜ 0.05).Conclusion The blood and sputum levels of leukotriene increase in children with RSV bronchiolitis,which is related with the severity of bronchiolitis.

Objective To investigate whether erythropoietin(Epo) is potentially beneficial in protecting cultured retinal neurocytes.Methods Primary isolated retinal nerve cells were cultured.Expressions of Epo and EpoR protein in cultured retinal neurocytes were decected by immunohistochemical analysis.Survival of cultured neurocytes that were incubated in the presence of Epo or glutamate in the presence or absence of Epo were estimated by determining the activity of their mitochondrial dehydrogenases using the MTT assay.Results Epo and EpoR protein were expressed on the cultured retinal neurocytes.The presence of different concentrations of Epo did not improve the survival of retinal neurocytes,and Epo could prevent glutamateinduced toxicity. Conclusion Epo is beneficial in protecting mixed cultured retinal neurocytes from glutamate-induced cytotoxicity.

Objective To validate and compare the pyramidal tracts traced by functional magnetic resonance mo?tion activated area as the region of interest method (fMRI guided DTI-FT) and by the anatomy of primary motor cortex as region of interest method(traditional DTI-FT)using subcortical electrical stimulation (DsCS). Methods A prospective study was conducted in 12 cases of patients with central lesions involving the motor area. The pyramidal tracts were traced by fMRI guided DTI-FT method and traditional DTI-FT method. The lesions were resected with the assistance of neuronavigation. The distances between same stimulation positive point and pyramidal tracts traced by the fMRI DTI-FT or traditional DTI-FT were recorded. The coincidence rates between pyramidal tract imaging and DsCS were analyzed in order to verify the accuracy and reliability of these two methods. Results Two cases were excluded:one due to the failure of the fMRI activation caused by movement dysfunction and one case due to negative electrical stimulation.,The pyrami?dal tracts were successfully reconstructed in the rest 10 patients using these two methods which were further applied to assist surgery. The coincidence rates between DsCS and pyramidal tracts were 77%in fMRI DTI-FT and 70%in tradition?al DTI-FT. The shortest distances were 4.3mm±2.8mm and 5.5mm±3.4mm in fMRI DTI-FT and in traditional DTI-FT in 16 DsCS positive sites and the difference was statistically significant (P<0.05). Five cases had temporary postoperative pa?ralysis. Among them, four cases had upper limb paralysis and one case had hemiplegia. The motor function was improved in four cases and remained unchanged in two cases two weeks after the operation. The motor function in the rest six cases did not have any change before and after operation. Conclusion The fMRI guided DTI-FT can be helpful to deal with le?sions and effectively protect the brain function area in patients with the central area lesions involved motor area.

Bronchial Neoplasms ; pathology ; surgery ; Humans ; Lung Transplantation ; Male ; Middle Aged ; Transplantation, Autologous

AIMTo detect the changes of inducible nitric oxide synthase (iNOS) expression in liver following ischemia/reperfusion (I/R) of hindlimbs and to elucidate their significance.

METHODSI/R was established using the occlusion of the femoral arteries for 4 h and reopening for 2-24 h in rats. The expression of iNOS mRNA, and iNOS protein and the nitrotyrosine (NT), a marker of peroxynitrite (ONOO-), in liver tissue were detected with reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical technique, respectively. The liver superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents were spectrophotometrically measured. The observation of pathologic changes of liver was made following the inhibition of iNOS by aminoguanidine (AG).

RESULTSCompared with control groups, the relative expression level of iNOS mRNA significantly increased in I/R group. There were more iNOS positive hepatocytes and more NT positive hepatocytes in I/R group than control groups. The contents of MDA markedly increased, while the activity of SOD significantly decreased in I/R group, compared with those in the control groups. The pathologic changes of rat liver became milder in I/R group following the inhibition of iNOS by AG.

CONCLUSIONThe expressions of iNOS mRNA and protein in liver are significantly upregulated, excess induction of iNOS-NO is contributed to the liver injury during the I/R of hindlimbs.

Animals ; Guanidines ; pharmacology ; Hindlimb ; blood supply ; Liver ; blood supply ; metabolism ; Male ; Malondialdehyde ; analysis ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; Superoxide Dismutase ; analysis

The purpose of this study is to explore the feasibility and superiority of using rapid expansion of supercritical solution (RESS) technology in the field of traditional Chinese medicine. The extract of magnolia bark (EMB) was obtained by supercritical carbon dioxide (SCF-CO2) extraction technology. Microparticles of EMB were manufactured by RESS technology. The effects of operating temperature and pressure on the contents of the active ingredient in the particles were evaluated by HPLC. The effect of expansion conditions on the particle size distribution of EMB particles was investigated. The smallest sample (mean size: 4.7 microm) was obtained under the RESS condition: pressure of 25 MPa, temperature of 50 degrees C and a nozzle size of 100 microm. The characteristics of microparticles were also studied by differential scanning calorimetry (DSC), scanning electron microscopy (SEM) and image analysis. The dissolution rate study showed that microparticles had a significantly faster dissolution rate than normal material particles. After oral raw EMB suspension, the mean areas under the plasma concentration-time curves (AUC(0-t)) of honokiol and magnolol were found to be (4.23 +/- 0.36) and (5.46 +/- 0.57) mg x h x L(-1), respectively, which were increased significantly, i.e. (5.41 +/- 0.63) and (7.24 +/- 0.83) mg x h x L(-1) when micronized EMB suspension was administered orally in SD rats (P < 0.05). Similarly, the mean maximum plasma concentrations of honokiol and magnolol increased from (1.55 +/- 0.22) and (2.35 +/- 0.14) mg x L(-1) (raw EMB) to (2.31 +/- 0.17) and (2.84 +/- 0.21) mg x L(-1) (micronized EMB), respectively. The results of t-test demonstrated that AUC(0-t) and Cmax value for honokiol and magnolol was significantly increased with the micronization compared to raw EBM (P < 0.05). This study demonstrated that the RESS was applicable for preparing microparticles of EMB at low operating temperature. The process was simple, free of environment pollution and without residual solvent.
Administration, Oral ; Animals ; Area Under Curve ; Biphenyl Compounds ; blood ; pharmacokinetics ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; pharmacokinetics ; Lignans ; blood ; pharmacokinetics ; Magnolia ; chemistry ; Male ; Microspheres ; Particle Size ; Plant Bark ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Solubility