Objective To construct antibody library in patient with pemphigus vulgaris using phage display antibody technique. MethodsTotal RNA was extracted from B cells of patients with pemphigus vulgaris(PV) using polymerase chain reaction,immunoglobulin VHand VL geneswere amplified by specific primer of HuVHBACK, HuJHFOR, HuVkBACK and HuJkFOR. The VH and VL genes were then cloned into a phage vector and expressed on the phage surface as a fusionprotein with product of gene Ⅲ.ResultsThe antibody phage library of single chain antibodies of patients with PV was constructed. ConclusionWe have obtained EC1-2 and EC3-4 purified protein of Dsg3 from constructed recombinant plasma in our laboratory which will facilitate the study on Dsg3 antibodies.

Objective To investigate the effects of targeted forkhead box E1( FOXE1) gene on epitheli-al-mesenchymal transition(EMT) of papillary thyroid cancer (PTC) cell line TPC-1, and to study its role in in-vasion and migration of TPC-1 cells.Methods The lentiviral expression vector for RNA interference of FOXE 1 gene was constructed to silence the expression of FOXE 1.Real-time polymerase chain reaction ( RT-PCR) and Western blot were used to detect the expression of such EMT markers as E-cadherin , N-cadherin and Vimentin . Transwell assay and Scratch assay were used to analyze TPC-1 cells migration and invasion .Results After RNA interference of FOXE1, the morphology of TPC-1 cells indicated a transformation of EMT .The expression of epi-thelial phenotype marker E-cadherin decreased remarkably while mesenchymal marker Vimentin was significantly up-regulated(P<0.01).Compared with the control group , the expression of Vimentin mRNA in the experimen-tal group was 2.24 times higher .The migration and invasion ability of TPC-1 cells increased significantly , and the number of cells in transwell was 2.11 times of the control's.Conclusion The silence of FOXE1 gene probably increases the invasion potential of human PTC cells through EMT .

AIMTo examine the liver mechanism with which clenbuterol (CL) is explained how to affect growth metabolism.

METHODSThe technique of chronic poly catheter was used to study the effects of CL (0.8 mg/kg b w) on the hepatic flux of nitrogen, VFA and glucose in 4 sheep.

RESULTSThe urea-nitrogen flux in CL-treated period always was lower than that in control during 24 h. The average flux of urea-nitrogen in hepatic and portal vein were decreased by 16.86% (P < 0.01) and 15.51% (P < 0.05), respectively, compared with that of control. The peptide level in hepatic vein was decreased with the treatment of CL, average flux of peptide was decreased by 38.71% (P < 0.01). But the peptide level of portal vein in CL treatment period was similar to control. Moreover, VFA level in the portal vein was enhanced by CL, the average flux of acetate in portal vein was increased by 19.49% (P < 0.01). No difference of VFA level in hepatic vein was noted between CL-treated period and control. In addition, the glucose flux in hepatic vein was obviously increased with CL treatment, the average flux of glucose was increased by 25.96% (P < 0.01). And glucose flux in portal vein was also elevated during CL-treated period.

CONCLUSIONCL can affect growth metabolism of animal with increasing nitrogen deposition, improving absorption and utilization of VFA and enhancing glucose synthesis in sheep liver.

Animals ; Clenbuterol ; pharmacology ; Fatty Acids, Volatile ; metabolism ; Glucose ; metabolism ; Liver ; drug effects ; metabolism ; Sheep

Objective To explore the changes of neutrophil elastase (NE) and surfactant protein A (SP-A) in acute lung injury(ALI) rats,and the effect of antioxidant. Methods Sixty healthy mature Wister rats were divided into 2 groups, the control group and treatment group. The rats in two groups all received peritoneal injection of E. coli to establish the ALI animal model. 30 minutes after injection of E. coli,the rats in treatment group were injected reduced glutathione from vena caudalis. The levels of NE in blood and expressions of SP-A in lung tissue were detected at 3,6 and 12 hours after injection of E. coli. Results ALI symptom appeared 3 hours after injection of E. coli in the control group, obvious after 6 hours, the rats vomi-ted pink secretion after 12 hours. Lung edema and bleeding were found by pathologic examination. No obvious symptom was found in treatment group after 3 hours, slight tachypnea after 6 hours, slight edema in pulmonary tissue after 12 hours. After administration of reduced glutathione,levels of NE at 3,6 and 12 hours in the treatment group were lower than those in the control group,and indicated statistical significance in 6 and 12 hours(P <0. 05) ;Levels of SP-A in 3,6 and 12 hours in the treatment group were higher than those in the control group, and indicated statistical significance in 3,6 and 12 hours (P < 0. 05). Conclusion Dysfunction of pulmonary surfactant is secondary in ALI, degradation of SP-A is the one of reasons, the application of reduced glutathione as antioxidant, could effectively suppress NE to decompose basosexine elastin of cells and destroy surface active protein, has protective effect on ALI.

A set of central composite design experiments were designed by using four factors which were ethanol amount, ethanol concentration, refrigeration temperature and refrigeration time. The relation between these factors with the target variables of the retention rate of schizandrol A, the soluble solids content, the removal rate of fructose and the removal rate of glucose were analyzed with Bayesian networks, and ethanol amount and ethanol concentration were found as the critical process parameters. Then a network model was built with 2 inputs and 4 outputs using back propagation artificial neural networks which was optimized by genetic algorithms. The R2 and MSE from the training set were 0.983 8 and 0.001 1. The R2 and MSE from the test set were 0.975 9 and 0.001 8. The results showed that network analysis method could be used for modeling of Schisandrae Chinensis Fructus ethanol precipitation process and identify critical operating parameters.
Bayes Theorem ; Chemical Precipitation ; Cold Temperature ; Cyclooctanes ; chemistry ; Ethanol ; chemistry ; Fructose ; analysis ; Fruit ; chemistry ; Glucose ; analysis ; Lignans ; chemistry ; Neural Networks (Computer) ; Polycyclic Compounds ; chemistry ; Reproducibility of Results ; Schisandra ; chemistry ; Time Factors

Objective To investigate the heat effects of the high-frequency electric knife on the recurrent laryngeal nerve ( RLN) in pigs and the safety margin in which electric knife can be used .Methods Totally 12 pigs for experiment were randomly divided into 3 groups by the distance between the head of the electric knife and the nerve(2 mm, 1 mm, 0 mm).The application time of the electric knife touching RLN was set to be 3 s and the application energy of the electric knife was 90 W.The data of electromyogram were measured by means of nerve detector before and after operation .Statistical analysis was made based on those data .The nerve tissues were taken to make paraffin sections so that the histological change can be observed and compared before and after damage.Results The data from electromyogram by nerve detector indicated that the difference of the amplitude of group 2(1 mm)and group 3(0 mm)had statistical significance(P<0.05)within group.There were significant difference of the amplitude between the 3 groups.Histological study showed that the tissues of group 3(0 mm)had obvious injury .Conclusions In thyroid surgery , the safety range of high frequency electrical knife used around RLN is:the distance from electrical knife head to nerve should be no less than 2 mm.

Objective To detect anti-clinically amyopathic dermatomyositis (CADM)-140 antibody in patients with dermatomyositis (DM) or CADM,and to estimate its clinical correlation.Methods Serum samples were collected from 22 patients with DM,16 patients with CADM,46 patients with other connective tissue diseases complicated by interstitial lung disease(including 8 cases of polymyositis,15 cases of systemic lupus erythematosus,5 cases of systemic sclerosis,6 cases of Sj(o)gren syndrome,6 cases of mixed connective tissue disease,6 cases of idiopathic pulmonary fibrosis),and 5 normal human controls.Enzyme-linked immunosorbent assay (ELISA) was performed with the recombinant melanoma differentiation-associated gene 5(rMDA)as a substrate to measure the anti-CADM-140 antibody in these serum samples.Clinical manifestations were compared between patients with anti-CADM-140 antibody and those without.Results The anti-CADM-140antibody was found in 43.8% (7/16) of patients with CADM and 9.1%(2/22) of patients with DM(P<0.05),but absent in the patients with other connective tissue diseases and in the normal human controls.A significant incroase was observed in anti-CADM-140 antibody-positive patients with DM/CADM in the incidence of cutaneous ulceration and necrosis,interstitial lung disease and rapidly progressive interstitial lung disease (8/9 vs.6.9%,P<0.01;9/9 vs.48.3%,P<0.01;5/9 vs.0,P<0.05),serum lactate dehydrogenase level(328.3±104.2 vs 241.1±100.3 IU/L P<0.05),erythrocyte sedimentation rate(40.8±23.1 vs.22.5±16.8 mm/1 h,P<0.05),high resolution computed tomography score(122.9±54.8 vs.70.0±59.8,P<0.05)compared with anti-CADM-140 antibody-negative patients with DM/CADM.The ereatine kinase level was significantly lower(156.3±260.8 vs.1806.2±3737.1 IU/L P<0.05)in anti-CADM-140 antibody-positive patients with DM/CADM than in anti-CADM-140 antibody-negative patients with DM/CADM,while no significant difference was noted in the positivity rate of antinuclear antibodies or incidence of malignancies between the antibody-positive and-negative patients with DM/CADM.Conclusions Anti-CADM.140 antibody not only is useful for the diagnosis of interstitial lung disease in patients with DM/CADM,but also may serve as a serum marker for rapidly progressive interstitial lung disease.Monitoring of serum anti-CADM-140 antibody might help to predict the progression of interstitial lung disease in patients with DM/CADM.

Objective To investigate the combination application of the intracranial buried electrode and electrical stimulation techniques in excising the epileptogenic zone in the central zone.Methods Seven patients with epileptogenic zone located close to or in the central zone of brain were recruited in the present study.The lone term ECoG monitoring and electrical stimulation of the codex were performed to identify the epileptogenic zone and the central zone of the brain after patients received intracranial electrode implants.The epileptogenic zone was excised with maximum preservation of the cen-tral zone.The patients were follow-up for 6 to 12 months,the outcomes were evaluated based on the Engel's scale and the Karnofsky(KPS)score.Results Seven patients did not experience any seizures and their Engei's and KPS scores were markedly improve after operation.Conclusions Intracranial buried electrodes and cortical electrical stimulation can guide the resection of epileptogenic zone in the central zone.Patients have no seizure and no serious dysfunction after operation and their quality of life was improved markedly.

Objective To investigate the curative effect of endoscopic thyroidectomy via breast areola approach.Methods The clinical data of 28 cases of endoscopic thyroidectomy via breast areola approach were retrospectively summarized.Results The mean diameter ofthe tumor Was 2.9(1.7～4.2)cm.The mean operative time Was 128(95～165) min.Pathologic data:adenoma in 15 cases,nodular goitar in 12 csses,papillary carcinoma indicated intraoperatively by frozen section in 1 case.One case occurred temporary hoarseness.No intraoperative and postoperative hemorrhea and convulsion after operation.Minor pain and discomfort in anterior chest continued for 7～19d in 18 cases,all without taking painkillers,satisfaction rate of beauty was 90 percent.Condusions Endoscopic thyroidectomy via breast areola approach is safe and feasible for patients with thyroid diseases,and has excellent cosmetic results.

Objective To explore the clinical significance of intraoperative neuromonitoring (IONM)of recurrent laryngeal nerve (RLN) during thyroid cancer surgery.Methods 200 patients undergoing thyroid cancer surgery from Oct.2011 to May 2012 were retrospectively reviewed.The 200 patients were divided into 2 groups:100 patients in group A with IONM and 100 patients in group B without IONM.Results Group A had less RLN exposure time than group B(10.5 vs 15.3 mins,P ＜0.01).Group A had shorter operation time than group B (78.5 vs 82.3 mins).The difference had no statistical significance (P ＞ 0.05).Transient RLN palsy occurred to 5 cases in group A and 11 cases in group B(P＜0.05).Permanent RLN palsy occurred to 1 case in group A.No one in groupB had permanent RLN palsy (P ＞ 0.05).Conclusions IONM can reduce the incidence of transient RLN palsy compared with visualization alone.It is an effective procedure for identifying and protecting RLN in thyroid cancer surgery.