The decoy receptor-3 ( DcR3 ) and glutamic acid decarboxylase-65 ( GAD65 ) recombinant adenovirus was construced and transduced into denlritic cells (DC). After the transduced DC were utilized to immunize NOD mice,the CD+8 T cells and blood glucose were analyzed. The results showed that recombinant adenovirus inhibited the proliferation and cytokine release of GAD65 specific T cells,and delayed the incidence of diabetes.Both interferon-γ[ (50.5±7.2)vs(95.4±6.9) and(91.2±6.5) pg/ml] and interleukin-2 [ (46.3±5.1 )vs ( 86.1 ±5.2 ) and ( 80.3 ± 7.3 ) pg/ml ] were decreased compared to those in negative and blank controls ( all P＜0.05 ).The results suggest that DcR3 and GAD65 recombinant adenovirus might provide a promising way for gene therapy of type 1 diabetes.

Objective To analyze the detection rate of HBV serological makers in non-hepatic inpatients in the past six years. Methods Serum samples of 70 582 non-hepatic inpatients from three large hospitals were collected during 2003 to 2008. Serological markers of HBV ( HBsAg, anti-HBs, HBeAg, antiHBe and anti-HBc) were detected by the AxSYM MEIA system (Abbott Laboratories,Abbott Park,IL).Combining the test results of serological makers with other clinical data, several analysis models for this retrospective study were set up to evaluate the year-to-year changes in serological makers and the detection rates of each model. Results The order from high to low of detection rate of the 5 HBV serological markers was anti-HBc (55. 17% ), anti-HBs (49. 57% ), anti-HBe (28.42%), HBsAg ( 8. 92% ) and HBeAg (2. 12% ), and all of them had a downward trend in the past six years. The positive rate of HBsAg went down from 9. 30% (2003) to 8.70% (2008). The positive rate of HBsAg among people who were born after 1992 (2. 28% ) were significantly lower than that of the overall population (8. 92% ) and fell from 3.57%(2003) to 1.85% (2008). Each detection rate of all serological makers had male sexual side effect [HBsAg ( 12. 38%/7. 25% ), HBeAg ( 2. 72%/1.58% ), anti-HBc ( 56. 57%/53.43% ), anti-HBe (41.50%/28. 35% ) and anti-HBs (65.48%/50. 00% ), male/female]. The differences were statistically significant (Chi-square values of HBsAg, HBeAg, anti-HBc, anti-HBe and anti-HBs were 509.74,105.78, 69.66, 1 321.61 and 1 726.91, respectively; all P ＜ 0. 01).Twenty-six models of HBV serological makers from 70 582 inpatients were summed up, and 8 models had positive rates geater than or equal to 1%. The "All Negative" model ranked No. 1 and had no significant change from year to year. During the past six years, models representing "A11 Negative" and "anti-HBs Positive alone" were mainly in individuals younger than or equal to 20-year-old, while the models representing "anti-HBc and/or anti-HBe,anti-HBs Positive" were mostly in people older than 20-year-old. The distribution curve of models representing "HBsAg, HbeAg and anti-HBc Positive" and "HBsAg, anti-HBc, anti-HBe Positive"etc. showed a bell-shape, covering the population from 20-year-old to 70-year-old. Conclusions The slowlydescending tendency of the detection rates of HBV serological makers was observed during the past six years.The detection rates of HBV in the younger generation decreased significantly. However, the HBV infection rates of overall population is still high, so it is a high time that we made continuous improvement for the serum HBV screening technique in order to reduce the HBV infection ratess.

Objective To study the effect of propofol intravenous anesthesia on T helper cells of patients with primary liver cancer during perioperative period.Methods A total of 86 patients with primary liver cancer in our hospital from November 2014 to October 2015 were selected,who were divided into observation group and control group according to the method of random numbers,43 cases in each group.The observation group were taken propofol intravenous anesthesia,and the control group were treated with sevoflurane inhalation anesthesia.The Th1 cells percentage,Th2 cells percentage and the ratio changes of the Th1/Th2 cells of two groups before anesthesia and postoperative 1 day were compared.The plasma cortisol levels of two groups before anesthesia,after anesthesia,intraoperative and 1 day after operation were observed.Results The percentage of Th2 cells in observation group and control group 1 day after surgery had no significant difference (P ＞ 0.05).The percentage of Th1 cells and Th1/Th2 cells ratio of the observation group were higher than those of the control group [(16.32 ± 1.76) ％ vs.(14.16 ± 1.03),(8.48 ± 0.92) vs.(7.11 ± 0.72)],the differences were significant (P ＜ 0.05).The plasma cortisol levels of observation group during operation and 1 day after operation were lower than those of the control group[(12.34 ± 1.02) μg/dL vs.(16.13 ± 1.26) μg/dL,(12.01 ± 0.94) μg/dL vs.(15.25 ± 1.08) μg/dL],the differences were significant(P ＜ 0.05).Conclusion The propofol intravenous anesthesia can encourage more Th to differentiate into Th1 cells,which plays a protective role in the patient's immune function.

Objective To investigate the analgesic effect of salmon calcitonin(sCT)and its effect on expression of calcitonin receptor(CT-R)in periaqueductal gray(PAG). Methods Rat models of neuropathic pain were prepared by chronic constriction injury(CCI). Thermal withdrawal latency(TWL)and mechanical nociceptive threshold(MNT)were measured using hot plate test and yon Frey monofilaments test. The distribution of CT-R in PAG was detected by immunohistochemical method. CT-R protein was quantitatively determined by western blotting. Fourty male SD rats were randomized into 5 groups: normal group, sham-CCI group, CCI group, CCI plussubcutaneous sCT group, and CCI plus microinjection of sCT into PAG group. Results TWL, MNT, andexpression of CT-R in PAG showed no difference between normal group and sham-CCI group(P＞0. 05). TWL and MNT in CCI group were significantly lower than those in normal group(P＜0.05), and expression of CT-R in CCI group was significantly higher than that in normal group(P＜0.05). TWL, MNT and expression of CT-R in CCI rats increased significantly after sCT therapy(P＜0. 05), and the effect was more marked in PAG injection group than subcutaneous injection group(P＜0.05). Conclusions sCT raises the pain threshold and increase the expression of CT-R in PAG of CCI rats, while PAG injection showed more marked effect than subcutaneous injection.

Objective To evaluate the effect of Critical Incident Reporting System on the quality of clinical anesthesia.Methods Anesthesia-related critical incidents happened in the perioperative period were reported in voluntary,anonymous,no punishment and confidential manners.The data was collected,classified and documented by assigned professionals on a regular basis from September 2012 to August 2016.The critical incidents were retrospectively analyzed after the risk was assessed.The 4-year reporting rate was collected.The risk of critical incidents was assessed using severity and probability analysis,and the critical incidents-inducing risk factors were analyzed.Results The 4-year reporting rate of critical incidents was 0.551％.From 1st to 4th year,the reporting rates were 0.729％,0.598％,0.819％ and 0.368％,respectively,and the incidence of injury incidents was 0.112％,0.106％,0.133％ and 0.031％,respectively.The reporting rate of critical incidents and incidence and reporting rate of the injury incidents showed a decreasing trend for 1st and 2nd year,significantly increased for 3rd year and decreased for 4th year (P＜0.05).The first three critical incident categories were equipment use and respiratory system-and workflowrelated incidents.Patient injury during surgery was considered an extremely high risk incident;the factor of the medical staff in the department of anesthesiology is the first critical incidents-inducing risk factor.Conclusion Critical Incident Reporting System can discover and correct the system-related risk and the inducing factors in the department of anesthesiology and is an effective method of improving the service quality of clinical anesthesia.

Objective To explore the molecular epidemiological characteristics of norovirus isolated from the patients with acute gastroenteritis in Zhejiang province during 2016.Methods The stool samples and clinical data of 1 308 patients with acute gastroenteritis were collected from January to December in 2016.The type Ⅰ and Ⅱ of norovirus in stool samples were detected by one-step double real-time RT-PCR.Some of the positive specimens were selected by stratified sampling and amplified by conventional RT-PCR,and the PCR products were sequenced for genotype identification and phylogenetic analysis.Results Among the samples studied the positive rate of norovirus was 10.55％ (138/1 308) in which 12 cases were GⅠ genotype,118 cases were G Ⅱ genotype and 8 cases were mixed infection of G Ⅰ/G Ⅱ genotypes.The positive rate of norovirus in different age groups decreased with the increased age of patients,and became the lowest in the patient group of more than or equal to 60 years old.There was no significant difference for the positive rates of norovirus in different genders.Norovirus infection was distributed throughout all the year with the peak value of positive rate (37.50％) in December.The sequence analysis demonstrated that G Ⅱ.4 and G Ⅱ.17 genotypes were the prevalent strains of G Ⅱ genotypes with proportions of 40.91％ (18/44) and 34.09％ (15/44),while GⅠ.6 genotype was the prevalent strain of GⅠ genotypes.Conclusion Norovirus should be the important pathogen causing acute gastroenteritis in Zhejiang province during 2016.G Ⅱ.4 and G Ⅱ.17 of norovirus may be the predominant epidemic genotypes.

Objective: To investigate the etiology composition of enterovirus (EV) in patients with severe hand, foot, and mouth disease (HFMD) in children. To assess the diagnostic value of cerebrospinal fluid (CSF) tests in severe HFMD, and to find the key laboratory tests for severe HFMD. Methods: A total of 288 hospitalized cases of children clinically diagnosed with severe HFMD in Hangzhou Children′s Hospital were included from March to July 2016. Throat swabs were collected and enterovirus nucleic acids were detected by fluorescence quantitative reverse transcription (RT)-PCR. Synchronous CSF and serum samples were collected for EV-A71 and CV-sackievirus A16 (CV-A16)-IgM antibody detection. CSF samples underwent routine and biochemical tests. Normally distributed continuous variables were compared using t test. Non-normal distribution continuous variables were compared using Mann-Whitney U test. Differences between categorical variables were compared with χ² test. Results: The total positive rate of enterovirus nucleic acid EV-A71/CV-A16/EV by fluorescence quantitative RT-PCR in the 288 cases of children clinically diagnosed with HFMD was 83.7% (241/288), including EV-A71 55.2% (159/288), CV-A16 4.9% (14/288) and the other enterovirus 23.6% (68/288). Among the other enterovirus group, there were 29.4% CV-A6 (20/68) , 16.2% CV-A4 (11/68) and CV-A10 10.3% (7/68). The total positive rate of combined serum and CSF detection of EV nucleic acid and EV-A71 and CV-A16 IgM antibody was 98.3% (283/288). EV nucleic acid positive rate was 83.7% (24/288). The positive rates were statistically different (χ² =37.289, P=0.000). The CSF nucleated cells count in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (Z=-4.472 and -9.991, respectively, both P<0.05). The CSF nucleated cells positive rate in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (χ²=43.857 and 133.078, respectively, both P<0.05). The CSF protein level in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (Z=-3.151 and -5.255, respectively, both P<0.05). The CSF protein positive (>400 mg/L) rate in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (χ²=4.956 and 11.795, respectively, both P<0.05). The CSF nucleated cell counts and positive rates in EV-A71 IgM antibody-positive subgroup were both higher than those in antibody-negative subgroup (both P<0.05). The CSF protein level and elevated proportion in antibody-positive subgroup were both higher than those in antibody-negative subgroup (both P<0.05). The lactate dehydrogenase concentration in antibody-positive subgroup was significantly higher than those in antibody-negative subgroup (P<0.05). The EV-A71 IgM antibody in serum was significantly correlated with the antibody in CSF (r=0.600, P<0.05). Conclusions EV-A71 is still the most important pathogen of severe HFMD in Hangzhou in 2016. Other enterovirus such as CV-A6, CV-A4 increases compared to those in 2014 and 2015. The CSF routine and biochemical tests and the IgM antibody levels can serve as an important indicator for the diagnosis of children with severe HFMD.

Objective To understand the changes on patterns of sleep duration of the China Health and Nutrition Survey (CHNS) cohort in 9 provinces from 2004 to 2011.Methods Four rounds of CHNS data were used.Urban/rural,age and gender specific insufficient sleeping rates and excessive sleeping rates were analyzed.Results In 2004,2006,2009 and 2011,a total of 274,281,329 and 304 children aged 3-5 years;874,806,768 and 742 children aged 6-12 years;789,529,426 and 367 children aged 13-17 years;9 568,9 530,9 942 and 9 609 adults aged ≥18 years were surveyed respectively.The lowest insufficient sleeping rate was 53.9％ (200/371) in 3-17 years old children in rural area in 2006,the highest insufficient sleeping rate was 77.2％ (44/57) in 3-5 years old children in urban area in 2004.The insufficient sleeping rate increased in rural 3-5 years old children from 2004 to 2011.For the adults aged ≥ 18 years,the insufficient sleeping rate ranged from 4.2％ (82/ 1 954) in females aged 18-44 years in 2004 and 2009 to 20.8％ (211/1 015) in urban residents aged ＞ 60 years in 2011.The insufficient sleeping rate in age-groups 44-59 years and ≥60 years increased in both males and females and in both urban area and rural area from 2004 to 2011.The gender specific excessive sleeping rate in 3-17 years old children was very low in both urban area and rural area and no difference was found in different rounds of survey.The excessive sleeping rate in adults ranged from 18.4％ (569/3 093) in urban population in 2011 to 32.5％ (1 617/4 969) in females in 2004.The excessive sleeping rate of adult decreased from 2004 to 2011.Conclusion We should pay attention to the fact that the insufficient sleeping rate in adolescents is high and in increase in rural 3-5 years old children and adults aged ≥45 years.

Objective: To establish a high performance liquid chromatography-tandem triple quadrupole mass spectrometry method for the detection of human plasma paraquat concentration. Methods: The plasma samples were pretreated with methanol to precipitate plasma protein, and then were separated by a Waters XBridge BEH HILIC column (2.5 μm, 2.1 mm × 100 mm) with acetonitrile-water containing 200 mmol/L of ammonium formate and 0.1% of formic acid as mobile phase and 0.4 mL/min of flow rate. The paraquat was monitored by ESI positive ion mode, multi-reaction ion monitoring (MRM) scanning, and m/z 186.1-171.1 as quantitative transition ion-pair. The plasma paraquat concentrations in patients were determined by the established method, and the clinical values of plasma paraquat concentration and severity index of paraquat poisoning (SIPP) were evaluated by the receiver operating characteristic (ROC) curve. Results: When the plasma paraquat concentration ranged from 50 to 10 000 ng/mL, the linearity was good (R 2 =0.997), and the lower limit of quantification was 50 ng/mL. The recovery rates and imprecisions of three quality control products at low (100 ng/mL), medium (2 000 ng/mL) and high (8 000 ng/mL) concentration levels all met the requirements, and no matrix effect was found. The pretreated samples were stable at room temperature for 6 hours, and the results were not affected by repeated freezing and thawing for 3 times. The SIPP of 31 poisoned patients was 17.76 (0.30-90.91) h·mg/L. The SIPP in dying patients was significantly higher than that in survival patients (P＜0.05). The area under the ROC curve of SIPP was 0.889, and the optimal cut-off value was 11.679 h·mg/L. Conclusion The established method is sensitive, accurate, rapid and specific, and suitable for the detection of plasma paraquat concentration in patients.

Influenza virus , the causative agent of influenza , has characteristics of fast transmission and strong infectivity, posing a major threat to public health .Currently, with widely use of the first-line antiviral neuraminidase inhibitors , drug-resistant strains have appeared and brought difficulties to the precaution and treatment of influenza .This article reviews influenza virus structural characteristics , drug-resistance mechanisms , resistance situation and detection methods of drug-resistant virus strains , aiming to provide a reference for future research on influenza virus drug-resistance and clinical medication .