A PCR method was used to amplify the sequence encoding the mature peptide of?-mannanase of Bacillus subtilis. The gene was inserted into the Pichia pastoris vector pPIC9K, downstream of?-factor signal peptide sequence. The resultant recombinant plasmid pPIC9K-MAN was lineared by BglII digestion and introduced into the host Pichia pastoris GS115 by PEG method. After screen, the recombinant P. pastoris strain MAN22 was obtained and fermented in large scale 5L fermenter. The recombinant mannanase activity could reach to 1102IU/ml. The properties of the recombinant mannanase were characterized.

It is still an important and hard work to diagnose anaphylactic shock in forensic practice. However, no breakthrough progresses in the diagnosis of anaphylactic shock and relevant research have been made so far due to the problems we used to meet in actual postmortem examination ,which are short of specific pathological changes in autopsy, the condition progress of patients who occur anaphylactic shock and history of allergy. Furthermore, patients suffer from diseases such as coronary heart disease, pneumonia, asthma, skin irritation, etc, and blood serum allergy biomarkers degrade after hemolysis on account of a long time from death to autopsy ,which are also the difficulties we have to cope with. The aim of this review is to focus on present situation and diagnostic index of anaphylactic shock including the pathological changes and some experimental methods such as special stain, immunohistochemical and serological test to provide reference for diagnosis and study of anaphylactic shock.

In order to improve the accuracy for quantitating the bovine foamy virus(BFV)in vitro,we developed a baby hamster kidney cell(BHK)-21-derived indicator cell line containing a plasmid that encodes the firefly luciferase driven by the BFV long terminal repeat promoter(LTR,from -7 to 1012). The BFV titer could be determined by detecting the luciferase expression since the viral trans-activator BTas protein activates the promoter activity of the LTR. One clone,designated BFVL,was selected from ten neomycin-resistant clones.BFVL showed a specific and inducible dose-and time-dependent luciferase activity in response to BFV infection.Although the changes in luciferase activity of BFVL peaked at 84 h post infection,it was possible to differentiate infected and uninfected cells at 48 h post infection. A linear relationship was established between the multiplicity of infection(MOI)of BFV and the activated ratio of luciferase expression in BFVL. Moreover,the sensitivity of the BFVL-based assay for detecting infectious BFV was 10,000 times higher than the conventional CPE-based assay at 48 h post infection. These findings suggest that the BFVL-based assay is rapid,easy,sensitive,quantitative and specific for detection of BFV infection.

BACKGROUND: The real mechanism of systemic sclerosis is still not clear, it is found clinically that there are psychological stress factors of different forms before the attack.OBJECTIVE: To deeply investigate the psychosocial factors in patients with systemic sclerosis, and primarily analyze the role of psychological stress factors by comparing with national norms and controls as well as combining with clinical immunological test.DESIGN: A controlled observation.SETTING: The experiments were carried out in the Department of Geriatrics, the First Hospital Affiliated to Medical College, Zhejiang University and the Department of Surgery, Hangzhou Tongji Hospital.PARTICIPANTS: Between December 2002 and September 2005. 26 patients with systemic sclerosis (systemic sclerosis group), who received thoracic duct lymph drainage therapy, and 30 inpatients with chronic gastritis(control group) were selected from the same disease area of the Department of Internal Medicine, the First Hospital of Medical College, Zhejiang University.METHODS: All the patients filled the general information inventory, the contents included were age, gender, main symptoms, disease course, special examination, diagnosis and drug therapy. The psychosocial scale was used to evaluate the psychosocial factors. Life event scale (LES) was used to assess the life events stress, including profession, learning, marriage and love, family and child, economics, justice, interpersonal relationship and other common life events. The simplified coping style questionnaire (SCSQ)was applied to assess the coping styles, including 8 main components: confrontation, indifference, self-control, seeking help, self-blame, escape, planning and reassessment, and then the habitual coping styles were divided into negative ones and positive ones. The Eysenck personality questionnaire (EPQ) was used to evaluate the personality characters, it consisted of4 subscales: extraversion-introversion scale, neuroticism scale, psychoticism scale, lie and cover up scale. The fasting blood samples (3 mL) were drawn from the patients to detect serum levels of immunoglobulin G (IgG), immunoglobulin A (IgA) immunoglobulin M (IgM) and complement C3 with immunoturbidimetry, and the correlations between IgG and other factors were analyzed.MAIN OUTCOME MEASURES: The evaluative results of psychosocial scale and results of clinical immunological detection were mainly observed.RESULTS: All the 56 patients finished the scale survey and immunological detection, and all were involved in the analysis of results. The total number of life events, number of negative events and LEU value of negative events in LES were all significantly greater in the systemic sclerosis group than in the control group. For the coping style, the dimension of positive coping was less but that of negative coping was more in the systemic sclerosis group than in the control group. For EPQ, the scores of extraversion-introversion were lower but the scores of neuroticism in both males and females in the systemic sclerosis group were higher than in the control group and norms. For the immunological detection, the levels of lgG, lgA and lgM were all higher in the systemic sclerosis group than in the control group, but C3 level had insignificant difference between the two groups.IgG had negative correlations with the number of negative events, dimension of negative coping and the score of extraversion-introversion in EPQ.CONCLUSION: Patients with systemic sclerosis have obvious psychological stress, negative coping style, unstable mood and abnormal humoral immune function. Psychosocial stress has influence on immunology, it is indicated that psychological stress is closely correlated with the attack of systemic sclerosis.

Objective: To evaluate the service competence of 99 smoking cessation clinics in Zhejiang Province, so as to provide reference for the improvement. Methods: The questionnaire, prepared by Tobacco Control Office of Chinese Center for Disease Control and Prevention, was used to investigate all the smoking cessation clinics in Zhejiang Province, to score the basic and developmental indicators, and to assess the qualified rates ( basic indicators >50 points and total scores >60 points ). Results: There were 99 smoking cessation clinics, with 52 in secondary hospitals and 47 in tertiary hospitals. The overall assessment scored from 11 to 96 points, with an average of ( 53.99±16.56 ) points. The smoking cessation clinics in secondary and tertiary hospitals scored ( 53.92±15.88 ) points and ( 54.06±17.45 ) points. The scores of basic and developmental indicators were ( 45.66±12.16 ) points and ( 8.33±5.39 ) points, with the scoring rates of 65.23% and 27.77%. The overall assessment of 43 smoking cessation clinics were qualified and the rate was 43.43%, which was 44.23% in secondary hospitals and 42.55% in tertiary hospitals. Conclusion The qualified rate of smoking cessation clinics in Zhejiang Province was 43.43%, which was similar between second hospitals and tertiary hospitals.

OBJECTIVETo investigate the value of Qingyi Decoction (QYD) and Glauber's salt (GS) for treatment of severe acute pancreatitis (SAP).

METHODSSixty-five patients with SAP were randomly assigned to two groups, the treated group (33 patients) and the control group (32 patients). They received the same therapy except that to the patients in the treated group, QYD was given through naso-gastric tube and GS was applied externally.

RESULTSCompared with those in the control group, the alleviating time of clinical symptoms, such as abdominal pain and distention, time of hospitalization and time of blood amylase recovery in the treated group were shorter, with lesser complications and lower rate of transferring to operation (all with P < 0.05), showing a better efficacy in all aspects, but the mortality in the two groups was not different (P > 0.05).

CONCLUSIONCombined use of QYD and GS has significant therapeutic effect for treatment of SAP.

Adult ; Aged ; Double-Blind Method ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Pancreatitis, Acute Necrotizing ; drug therapy ; Phytotherapy ; Prospective Studies ; Sulfates ; therapeutic use ; Treatment Outcome

Objective To investigate the value of the application of the fresh first morning midstream urine in cytological study of bladder cancer patients. Methods The results of the fresh first and second morning midstream urine cytological studies for 52 bladder cancer patients were analyzed.Continual three urine samples and single urine sample were treated as study objects respectively. The positive rates in different tumor stages and grades were evaluated. Results The positive rate of overall 52 patients was 78. 8 % (41/52) in fresh first morning midstream urine and 80. 8% ( 42 / 52) in the fresh second morning midstream urine. While in 156 single urine samples, the positive percentages were 56.4%(88/156) and 60. 9% (95/156). The positive rates of the fresh first and second morning midstream urine were 69.7% (23/33) and 72.7% (24/33) respectively in grade 1- 2 patients, and 44.4 % (44/99) and 48. 5 % (48/99) in 99 single urine samples. The positive rates of 42 non-muscle invasive bladder cancer patients were 73. 8% (31/42) and 76.2% (32/42) in the fresh first and second morning midstream urine, while in 126 single urine samples, the positive rates were 54.8% (69/126)and 57.1% (72/126). There were no significant differences between positive rate of the fresh first and second morning midstream urine in diagnosis of bladder cancer, low grade bladder cancer and nonmuscle invasive bladder cancer. Conclusion The fresh first morning midstream urine can be used for urine cytological study in the diagnosis of bladder cancer, even in the diagnosis of low stage and low grade bladder cancer.

The quality markers (Q-markers) of traditional Chinese medicine (TCM) have become a topic of interest in TCM research in recent years. Nonetheless, there is still no consensus on how to scientifically characterize TCM Q-markers. Our study establishes an identification method for TCM Q-markers based on the analytical hierarchy process (AHP) and the entropy weight comprehensive method. By constructing an evaluation system encompassing the target layer, the factor layer and the control layer, AHP can be used to analyze the weight of three core TCM quality attributes, including effectiveness, testability and specificity. Following that, the entropy weight method is employed to analyze the specific indicators for each attribute based on the literature and experimental data. Finally, the comprehensive weight of each index is obtained by combining the two weights, and the comprehensive weight and the specific value of each component is multiplied and summed to obtain the integrated score ranking, and thereby identify the TCM Q-markers. Taking Shaoyao Gancao decoction as an example, the analysis revealed that the top 8 components are as follows: paeoniflorin > quercetin > albiflorin > glycyrrhizic acid > naringenin > liquiritin > oxypaeoniflorin > benzoylpaeoniflorin, and can be identified as Q-markers of Shaoyao Gancao decoction. This study not only provides support for the establishment of quality standards and process quality control of TCM formulae, but also provides innovative ideas and methods for quantitative evaluation and accurate identification of TCM Q-markers.

Objective To evaluate the use of gated SPECT in patients with hypertrophic obstructive cardiomyopathy (HOCM) and the effects of percutaneous transluminal septal myocardial ablation (PTSMA) on myocardial perfusion.Methods 99 Tcm-methoxyisobutylisonitrile (MIBI) and 18F-fluorodeoxyglucose (FDG) images were performed in 31 HOCM patients before PFSMA and in 15 patients 3-7 d after PTSMA.The images in different left ventricular segments were analysed by using scores.Results In 99Tcm-MIBI images, uptake decreased at the septal regions in 12 HOCM patients (80.0%, 12/15) after PTSMA, 18F-FDG images also showed decreased uptake at the septal regions in 5 HOCM patients (33.3%, 5/15) after PTSMA.Conclusion 99Tcm-MIBI images might be an important method to evaluate PTSMA results, and 18 F-FDG images showed important value as reference.

OBJECTIVETo investigate the psychological impact of mastectomy on women with breast cancer.

METHODSQuestionnaires were answered by 90 patients after mastectomy for breast cancer. The data were collected and analyzed regarding the age, occupation, education, income and living place of the patients. Their psychological changes were compared and evaluated statistically using chi-square analysis.

RESULTSMastectomy caused psychological impact on half of the patients, which included fear of cancer, feeling of body incompleteness, inconvenience in working and social communication, less sexual act and low spirit. 79 percent of the patients considered breast reconstruction unnecessary. 54 percent did not know that the breast could be reconstructed. The patient feelings differed with the age, occupation, income and living place.

CONCLUSIONMastectomy hurts patient psychological health.

Age Factors ; Body Image ; Breast Neoplasms ; psychology ; surgery ; Chi-Square Distribution ; Fear ; Female ; Humans ; Mammaplasty ; psychology ; Mastectomy ; psychology ; Socioeconomic Factors ; Surveys and Questionnaires