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.

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.

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

It is the objective of this study to develop dynamic predictive model for the extraction process of red Ginseng using NIR spectroscopy. NIR spectroscopy was collected online and PLSR models were developed for total quantity of ginsenosides. The performance of NIR prediction model achieved R, RMSEC, RMSEP of 0.996 09, 0.018 9, 0.016 8, respectively. A first order dynamic mass transfer model was combined with NIR prediction of the quality indicator to predict the trajectory of the extraction process based upon the initial 3 or 4 data points. The results showed good agreement with actual measurements indicating reasonable accuracy of the predictive model. It could potentially be used for advanced predictive control of the extraction process.
Chemical Fractionation ; methods ; Ginsenosides ; chemistry ; isolation & purification ; Models, Theoretical ; Panax ; chemistry ; Spectroscopy, Near-Infrared

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.

ObjectiveTo explore the clinic symptom and the characteristics of video,tightly close,intracranial electroencephalogram (EEG) of patients with central region diastematia epilepsy. Methods Retrospective analysis of 9 patients with central region diastematia epilepsy admitted from June,2007 to August,2009.The characteristics of all patients' seizure symptom and EEG manifestation were analyzed using patients' medical history,video and EEG records.ResultsPatients with central region diastematia epilepsy had relatively long sezure history.The duration of seizure was commonly short,with frequent episode and no obvious intelligence impairment.The seizure was often accompanied with the hyperkinesia in the lower limbs.Scalp EEG showed discharges with low amplitude waves in the mean line area.The superhigh amplitude and regular rhythm slow sharp wave could be found in the diastematia cortex EEG.All patients had an Engel Class Ⅰ outcome after surgery.ConclusionThe seizure symptoms are characteristic in the patients with central region diastematia epilepsy,and some special manifestations can be found in different phase,wave amplitude,rhythm,lead array.