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 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.

Recurrent laryngeal nerve injury is one of the serious complications after thyroidectomy.Unilateral injury causes hoarseness,while bilateral injury causes difficulty in breathing or even life-threatening glottis obstruction.Analyzing the root cause of the thyroid injury,firstly it is the anatomical factors of recurrent laryngeal nerve,namely the close and complex relationship between the recurrent laryngeal nerve and the inferior thyroid artery,the existence of branches of recurrent laryngeal nerve and its variation,and the presence of non recurrent laryngeal nerve and Zuckerkandl nodules.Those are all made the recurrent laryngeal nerve easy to be damaged.Secondly it is because of the vulnerability of the recurrent laryngeal nerve itself.Last improper using of energy operation instrument will cause heat injury on nerves.Below counter measures can be implemented to prevent recurrent laryngeal nerve injury.Dissect to show recurrent laryngeal nerve or make it ‘ visualization’ during thyroidectomy.Elaborately anatomize recurrent laryngeal nerve to appropriate degree.Be familiar with the property of energy operation instrument and thus safely use them to reduce the heat injury of recurrent laryngeal nerve.Reasonably use the intraoperative nerve monitoring in the surgery,which assist to reduce the risk of injury of recurrent laryngeal nerve.

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 optimize the extraction process of red ginseng to minimize the unit cost of extracting effective ingredients. The relation between the target variables of total quantity of ginsenosides and first extraction time, first extraction solution amount, second extraction time, second extract solution amount were studied with Box-Behnken experimental design method. At the same we also considered the cost of extraction solution and energy usage. The objective function was set as unit cost of target (total quantity of ginsenosides or its purity) for the multi-objective optimization of extraction process. As a result, the optimal process parameters were found as first extraction time (108.7 min), first extraction solution amount folds (12), second extraction time (30 min), second extraction solution amount folds (8) to minimize the unit cost. It indicated that this approach could potentially be used to optimize industrial extraction process for manufacturing Chinese medicine.
Chemistry, Pharmaceutical ; economics ; methods ; Cost Control ; Drugs, Chinese Herbal ; analysis ; economics ; isolation & purification ; Panax ; chemistry

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 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.