Objective To study the prokaryotic expression, puriifcation and properties of recombinant human Chymotrypsin. Methods The protein was highly expressed in E.coliBL 21 (DE 3) as inclusion body. After refolding and activated with trypsin, the activated protein was obtained and purified with ion-exchange chromatography(CM-FF), some properties of the recombinant human chymotrypsin was investigated. Results The molecular weight of chymotrypsinogen was about 30 KD in SDS-PAGE, total activity recovery rate of CM-FF puriifcation was 93.7%. The recombinant chymotrypsin kept stable from pH 3 to pH 5, and owned good temperature stability. Km was 0.067 mmol/L with BTEE as a substrate. The UV maximum absorption wavelength was 281 nm.Conculsion The recombinant human enzyme was expressed successfully, and a feasible production method to get a high activity of the recombinant human chymotrypsin was established.

Objective To investigate the distribution and environmental pollution of dioxin-like polychlorinated biphenyls(PCBs) in a place of disused transformer and other electronic waste disassembled. Methods The ambient air, aerosol, water, soil and vegetable samples were collected to analyze the dioxin-like PCBs level with isotope internal standards and HRGC-HRMS techniques. Results The concentration of dioxin-like PCBs in ambient air was 4 031.77-4 942.34 pg/m3(namely 1.832-2.666 pg TEQ/m3), while in aerosol it was 325.45-3 499.77 pg/m3(0.370 9-1.536 3 pg TEQ/m3). In the soil sample, the value was 2 316.59-19 100.30 pg/g(1.358-15.264 pg TEQ/g) dry weight. In water it was 5.731-237.640 ng/L(2.585-73.046 pg TEQ/L). In vegetable it was 1 078.45-1 620.04 pg/g(0.778 3-1.126 2 pg TEQ/g) wet weight. In all air, aerosol, soil, water and vegetable samples, the content of PCB-118 was the most. PCB-126 had the highest TEQ value. In non-ortho PCBs, PCB-77 was of maximum content. In mono-ortho PCBs, PCB-118 was the most, and the followings were PCB-105, PCB-156 and PCB-167. All the 12 kinds of PCBs were investigated in all samples except PCB-169 absent in ambient air. Conclusions The investigated area around the disassembly of obsolete solid waste has been badly contaminated by dioxin-like polychlorinated biphenyls(PCBs), and PCBs already exist in the vegetables. According to the results of the present paper, some further studies should be carried out to evaluate the toxic effects on food and people health.

Objective:To evaluate the reliability and validity of Emotional Problems Factor of Chinese Personality Assessment Inventory II(CPAI2-E).Methods:Item analysis of the test was carried out;internal consistency reliability, retest reliability and structure validity, differential validity were examined.Results:①Through factor analysis, 20 subordinate factors were got.②The Cronbach ? coefficient of the total was 0.948 and each factor ranged from 0.755 to 0.896;re-test reliability of the total test was 0.718.Conclusion:CPAI2-E can be used as an effective and reliable tool for assessing anxiety disorders.

Objective: To explore the relationships of personality characteristics, Anxiety trait, and general self-efficacy with anxiety disorders. Methods: By case-matched study, 144 patients with anxiety disorders and 144 normal controls were administered the Emotional Problems Factor of Chinese Personality Assessment Inventory II (CPAI2-E), State-Trait Anxiety Inventory and General Self-efficacy Scale. Results: ①The scores of patients with anxiety disorders were higher than those of the normal controls in CPAI2-E (P

Objective To construct a fusion minigene expression vector of seven HCV NS3 epitopes and express the fusion gene in the eukaryotic cell line, in order to form a foundation for the investigation of the DNA immunization for HCV prevention. Methods Three pairs of complementary oligonucleotides primers covering all seven HCV NS3 epitopes were synthesized. Overlapping extension PCR were performed to construct the fusion minigene and cloned in pEGFP-N3 and pBuDCE vector respectively. The fusion protein was detected by Western blotting and flow cytometric analysis. The transfection efficiency was assessed with by flow cytometric analysis. Results The HCV NS3 epitope fusion minigene was obtained and inserted into pEGFP-N3 and pBuDCE vector respectively. The recombinant plasmid pEGFP-DR4 and pBuDCE-DR4 were expressed in 293T and 046W cell lines respectively. The Western blot result indicated that the expected 36 kDa minigene/EGFP fusion product was expressed from pEGFP-DR4, while 13kDa product from pBuDCE-DR4 respectively. Conclusion The HCV NS3 epitope fusion minigene was expressed in the eukaryocyte expression system.

Objective:To investigate and compare the effect and the possible mechanisms of 2 hypoxia-inducible factors(HIF) 1? and 2? on renal cell carcinoma(RCC) by RNA interference(RNAi) method.Methods: Chemically synthesized HIF-1? and HIF-2? RNAi were transfected into RCC cell line A498 by liposome transient transfection.Normal proximal tubule cell line HK-2 and A498 were separately treated with medium and liposome as controls.The expressions of VEGF,ET-1,bcl-2 and Ki67 were observed by RT-PCR or Western blot 24 h after RNAi.Cell apoptosis was detected by flow cytometry and fluorescence microscopy.Results: Expressions of HIF-1? and HIF-2? RNAi were higher in A498 than those in HK-2,and the 2 genes were successfully blocked after RNAi.HIF-1? RNAi resulted in a significant decrease of ET-1,bcl-2 and Ki67 expression,with increased apoptosis and necrosis of cells.HIF-2? RNAi only resulted in a decrease of VEGF.Conclusion: Both HIF-1? and HIF-2? play important roles in regulation of angiogenesis; moreover,HIF-1? might also regulate proliferation of tumor cells,possibly through regulating anti-apoptotic genes.It is implied the existance of different transcription regulation functions and gene combining sites between HIF-1? and HIF-2?.

Objective To observe the therapeutic effects of oxygen inhalation combined with relaxation training on patients with generalized anxiety disorder (GAD).Methods A total of 188 GAD patients were divided into the study group and the control group randomly.The two groups were treated with conventional antianxiety drugs (paroxetine),in addition to conventional medical therapy.The study group was aided with oxygen inhalation and relaxation training,the control group was given benzodiazepine.The scores of Hamilton anxiety rating scale (HAMA),global improvement(GI) [ a part of clinical global impression (CGI) ],and treatment emergent symptom scale (TESS) were used to evaluate clinical effects and adverse reactions before and after 4-week treatment.The indexes of breathing,heart rate and blood pressure were evaluated before and after treatment.Results The scores of HAMA scale and GI in the two groups were significantly different after treatment( P ＜ 0.05 ),and the study group changed significantly( P ＜ 0.05 or P ＜ 0.01 ). The adverse reactions of the two groups were slight,there was no significant difference of TESS before and after treatment ( P ＞ 0.05 ). There was no significant difference in terms of breathing,heart rate and blood pressure between the two groups before treatment( P ＞ 0.05 ),but were all lower after treatment the study group changed more apparently and significantly(P ＜ 0.01 ).Conclusions Based on conventional drugs treatment oxygen inhalation combined with relaxation training could improve anxiety and somatization rapidly,there was no obvious adverse reaction and the patients compliance was good,so the treatment was worth to use in clinic.

‘Controlling spirit’is the key point of efficacy of acupuncture treatment, which goes throughout the treatment. Both doctor and patient have to understand each other and closely cooperate to get full control of spirit. Try to be carefully to examine the condition and get psychological relaxation before inserting needle, concentrate the spiri and get mind in the needle on inserting needle, give attention to needling sensation after inserting needle, setting the mind to conserve energies and regulate emotion-thought after witherawing needle.To achieve controlling spirit includes understanding spirit, influencing spirit, focus on spirit, and maintaining spirit, then to improve the clinical curative effect and obtain the the best results.

Objective The stability and other characteristics of the active recombinant human anionic trypsin(hT 2) with site-mutation R 122 L(mhT 2) were investigated. Methods An active human anionic trypsin and its R 122 L mutate were produced with E.coli BL 21(DE 3) and purified with ion-exchange chromatography. The properties of mutant were studied and compared with the wild type. Results The optimal pH for mhT 2 was 7~11. mhT 2 was active over a broad temperature range (4℃~80℃) and owned a little better thermal stability than the wild type. The inhibition of typical metal chelating agent(EDTA), Fe 3+, denaturant, reducer(β-ME) on activity of mhT 2 was the same as the wild type. Michaelis constant Km of mhT 2 was 0.010 mmol/L with BAEE as a substrate, a little lower than wild type. Conclusion Compared with the wild type, the R 122 L site mutate significantly enhanced tolerance to acidic pH、denaturants、reductions and autolysis.

Objective To establish a method for detecting chimerism after allogeneic stem cell transplantation using microsatellite polymorphism. Methods DNA was extracted from bone marrow or peripheral blood of 10 recipients and their donors, DNA fragments including 5 microsatellite loci were amplified by PCR, the polymorphism of PCR products was analyzed by PAGE and sliver staining, and the quantitative analysis of chimerism was performed using image analysis software. Results [WTBZ]The five selected STR loci had high polymorphism, and were suitable for detecting chimerism. The sensitivity of sliver staining was 90%. Conclusion Microsatellite polymorphism analysis based on PCR is a sensitive and accurate method to detecting chimerism after allogeneic stem cell transplantation, but the sensitivity of sliver staining was not so good,so more sensitive methods should be used.