Objective To establish immortalized neural progenitor cell strain and provide stable cell resource for cell-transplantation and gene therapies. Methods Plasmid pCMVSV40T/PUR containing the simian virus 40 large T antigen gene (SV40Tag) were transfected into the primary cultured neural progenitor cells (NPCs) of newborn rat using lipofectin transfection method. Colonies were isolated by puromycin selection and expanded by many passages. Anti-nestin antibodies were used to identify the cultured cells. The specific molecular marker of neurons and astrocytes were detected using immunocytochemistry method to investigate the capability of differentiation of the transfected cells. The expression of SV40Tag in expanded cell lines was identified by RT-PCR, Southern blot and immunocytochemistry method.Results One anti-puromycin cell clone was obtained, which was microtubule-associated protein-2 (MAP-2) positive cells with the capability of proliferation and could differentiate into MAP-2 or glial fibrillary acidic protein positive cells. The existence of SV40Tag cDNA and the expression of mRNA and protein of SV40Tag were confirmed in transfected cells. The transfected cells were expanded to immortalized cell strain maintained for more than 50 passages, named as immortalized neural progenitor cell (INPC) . INPCs were elliptical or triangular cells with two or three short axons. The population doubling time of INPC was (22.9?2.7)h, subculture, freezing and recovering had no effect on cellular shape and proliferation of INPC. Conclusion Immortalized neural progenitor cell strain was established successfully. It may provide stable cell resource for the basic researches and cell-transplantation therapies with NPC.

Objective To establish a method for the cultivation of hain follicle outer root sheath (ORS) cells of murine vibrissa and to study their expression of uPA/uPAR. Methods The ORS cells of murine vibrissa were cultured by combination of enzyme digestion and tissue culture with the feeder layer of dermal sheath cells of the hair follicle. The cells and their uPA/uPAR expression were identified by immunocytochemistry (ICC). Results The feeder layer of DS was suitable for the culture of ORS cells. ORS cells expressed uPA/uPAR protein. Conclusion The DS of hair follicle is a better feeder layer for the growth of ORS cells. ORS cells have the ability to migrate and proliferate.

Environmental physiological method adopted, the ergonomic experiment is to seek the best parameter of the protective hood structural size, on which the structure design of the protective hood will be based, through the analysis of the organic relationship between the microenvironment of power air-purifying medical protective hood and the body's physiological characteristics. The changes of such indexes due to different air currents are analyzed as the oxygen, carbon dioxide, temperature and humidity in the microenvironment. The changes of testees' physiological parameters and their responses to psychological loads and body strengths are paid attention to. The results show the hood can meet the testees' demands with proper air current.

Objective To explore the roles of urokinase type PA (uPA) and uPA receptor (uPA R) in the course of repair after human embryo corneal alkali burn in vitro . Methods After the alkali burn model in vitro was established successfully, some techniques such as the observation of cell culture and morphology, ICC and image analysis were used. Results No significant difference was found between the cells from corneal limbus and central cornea and almost all of them expressed AE1/AE3. The expressions of uPA and uPA R increased to the maximum at about 24 h after alkali burn. uPA expression distributed mainly in the cytoplasm but uPA R expression distributed mainly in the cell membrane. Conclusion The corneal epithelial cells of comparatively high purity can be acquired by tissue culture. There probably exists difference in development and vitality between embryo and adult cornea. There might be commonness of the roles of uPA and uPA R in epithelial tissue.

Two hundred and fifty patients with subclinical hypothyroidism ( SCH) and 120 euthyroid volunteers were recruited for the study, SCH patients were stratified into 2 groups according to TSH levels(group A:TSH<10 mIU/ L; group B: TSH>10 mIU/ L). All subjects were examined for clinical characteristics, thyroid profile, lipid profile, and biomarkers of early atherosclerosis. Patients in group B received L-thyroxine replacement treatment to achieve euthyroidism. After 6 months of stable euthyroidism all measurements were repeated. SCH patients had higher levels of total cholesterol(TC), low-density lipoprotein-cholesterol(LDL-C), asymmetric dimethyl arginine (ADMA), carotid artery intima media thickness(CIMT), thromboxane B2(TXB2), and C-reactive protein(CRP) but with lower nitric oxide(NO) level compared with euthyroid subjects. Levels of TC, LDL-C, CIMT, TXB2, and ADMA decreased, and NO level increased significantly after 6 months of euthyroidism. TSH was positively correlated with TC, LDL-C, CIMT, ADMA, and TXB2; and negatively correlated with NO. Based on multivariate regression analysis, TSH was an independent influential factor of CIMT and NO. We conclude that raised TSH is an important risk factor of atherosclerosis in SCH.

Objective To investigate the ligand-binging characteristics of Veli3 PDZ.Methods Random peptide library was screened using yeast two-hybrid method with Veli3 PDZ as bait.In combination with bioinformatics method all the potential ligands in human proteome were predicted by searching human databases with the consensus-binding sequences.Fourteen native human proteins predicted as ligands were chosen by their cellular locations and biological functions for validating protein interaction in yeast two-hybrid system.Results The C-terminal consensus sequences for the Veil3 PDZ binding is X-COOH(X denotes any amino acid),which indicates that Veli3 PDZ belongs to classⅠ.Six of fourteen native human proteins predicted as ligands were confirmed to be positive in the yeast two-hybrid system.There were a lot of interactions between PSD-95,another PDZ protein,and the ligands of Veli3 PDZ reported previously and discovered in this study.Conclusion The six novel potential ligands of Veli3 found in this study provide significant clues for discovering biological functions of Veil3 proteins.Moreover,Veli3 PDZ and PSD-95 PDZ may compete in binding the same ligands.

Objective To construct the eukaryotic expression plasmid of pEGFPC1uPAR gene and explore the effect on the proliferation and invasion ability of Pam 212 cells. Methods The human uPAR cDNA was cloned by PCR, and inserted into the eukaryotic expression plasmid pEGFPC1. After identification of sequencing, the reconstructive plasmid was transformed transiently into Pam 212 cells, then the cell growth and the invasion ability were evaluated. Results The reconstructive plasmid of pEGFPC1uPAR was validated by sequencing. The reconstructive plasmid can promote the growth of Pam 212 cells and enhance the invasion ability. Conclusion The pEGFPC1uPAR plasmid was constructed successfully and uPAR was confirmed to promote the growth and the invasion ability of Pam 212 cells, which lay the foundation for further studies of uPAR in vivo.

Objective To investigate the risks and benefits of different surgical treatments for cystic neoplasms of the pancreas (CNP).Methods The clinical data of 243 CNP patients were reviewed retrospectively.Different surgical treatments were adopted according to the site,size and invasiveness of the tumors.A long term follow-up was carried out for patients with small benign CNP,and a surgical excision is proposed if tumors progressed during the observation.Results 58 outpatients with no evidence of malignancy was followed up and had long-term survival,in which 4 patients received a surgical resection in case of tumor progression,and all of them were confirmed benign tumors.185 cases received surgical treatments,with a resection rate of 97.3％ (180/185),including 127 non-invasive tumors,and 58 cases of invasive tumors.Perioperative mortality was 2/185,and morbidity rate was 41/185.Pancreatic fistula was the most frequent complication,which was significantly associated with tumor site and excision extension.All patients with non-invasive CNP acquired a long term survival after surgical treatments.The postoperative 1,3,5 year survival rates for patients with invasive lesions were 89.6％,52.1％ and 29.2％,respectively.Conclusions Long term follow-up and observation is feasible for asymptomatic patients with benign CNP.A radical resection should be performed for malignancy,and a combined multi-organ resection may improve the prognosis for local advanced tumors.

Objective:To explore the psychological and behavioral pattern of state speech anxiety of participants of college speech contest under two conditions.Methods:Narrow band measurements of both Subjective Uncomfortable Degree(SUD)and objective behavioral assessments were used under two conditions.Results:Under lab condition,SUD rat-ings differed among four time bands,while the behavioral ratings failed to show any difference.Under field condition,for the prepared theme speech,SUD ratings differed among four time bands while they failed to show any difference for the improvisational speech.For both speech forms,several behavioral items displayed a pattern of habituation.Conclusion:Under both conditions,the psychological and behavioral pattern of participants' state speech anxiety display a pattern of habituation while the behavioral pattern is affected more by different conditions.

ATP-binding cassette protein E (ABCE1) has been annotated as an Rnase L inhibitor in eukaryotes. Previous study showed that the overexpression of ABCE1 was related with the occurrence and clinical stage of lung adenocarcinoma. As an initial investigation into the novel functions of ABCE1, siRNA-expressing vectors targeting sites of the ABCE1 gene were constructed from RNAi-Ready pSIREN-DNR-DsRed-Express vector. Cultured 95-D and NCI-H446 lung carcinoma cells were transfected with the siRNA-expressing vectors using FuGENE 6 and transfection efficiency was determined by using fluorescence microscopy. The expression level of ABCE1 protein was determined by Western blot and immunofluorescence staining. Cell viability was determined by MTT, cell cycle was analysed by flow cytometry.The apoptotic rate was observed by ELISA. Fluorescence microscopy showed a satisfactory transfection efficiency which was about 42.70%. Cell viability and the growth fraction were markedly suppressed, whereas the apoptosis was significantly increased in SiRNA-95-D and SiRNA-NCI-H446 cells than controls(P< 0.05). It can be concluded that the siRNA targeting ABCE1 gene shows a dramatic inhibitory effect on RNA transcription and protein expression and a promoting effect on the apoptosis in 95-D/NCI-H446 cells, which offers a reliable base for the further in vivo experiment.