Objectives To understand the epidemiological characteristics of Japanese Encephalitis in Shanghai and to provide evidence for preventing JE.Methods Epidemic characteristics,JEV antibody in healthy population and swine infection rate in Shanghai were analyzed by methods of field survey,serology and molecular biology.Results JE incident rate in Shanghai was 0.077/100,000 in 2006;and 0.129/100,000 in 2007.Antibody positive rate before JE epidemic fastigium was 60.39%;postive rate after epidemic was 85.44%.JE IgG positive rate was 26.92% in 3-month swine and 14.86% in swine for sale;JE Gene in mosquito was analyzed for type 1.Conclusion The JE prevalence rate is relatively low in Shanghai.JE antibody positive rate is high in Shanghai population.Swine as a media is infected by JE virus.The JE virus in mosquitoes belongs to genotype 1.

Expression conditions of induction strategies for the cytoplasmic inclusion bodies (IBs) production of liver targeted interferon IFN-CSP by recombinant Escherichia coli (E. coli) BL21 (DE3) were optimized in shake-flask cultures in this study. The factors of the optimized protocol included in the present study were pH, inducer IPTG (isopropyl beta-D-thiogalactoside) concentration, culture growth temperature, incubation time and induction point. The effects of those factors were investigated by 'single variable at a time' method, aimed to analyze characterization of the recombinant strain. Orthogonal experimental design was further used to optimize the above critical factors for IFN-CSP production. According to the expression optimization result, it was confirmed that the main influence factors were cell density and induction temperature. The IFN-CSP gene expression optimized conditions were: pH value of the culture medium was 6.0, culture temperature 37 degrees C, adding IPTG to final concentration 0.4 mmol/L when the recombinant strain growth density OD600 achieved 0.8 and induction time 4 h. At this point, the IBs represented 74.3% of the total cellular protein. Compared with the non-optimized condition, IFN-CSP production obtained in optimized induction strategies were increased by approx. 1.2-fold. The optimized induction strategy yielded 688.8 mg/L of IFN-CSP, providing experimental data to study the biology activity and productive technology of IFN-CSP.
Biotechnology ; methods ; Cell Culture Techniques ; methods ; Culture Media ; chemistry ; Escherichia coli ; metabolism ; Gene Expression ; Interferons ; biosynthesis ; Liver ; Temperature

The peptidyl-prolyl imide bond cis/trans isomerization of Xaa-Pro motif in the peptide and protein plays an important role to influence their conformation and function. Here, a series of model peptides including phosphorylated and its unphosphorylated counterparts were designed and synthesized. Preliminary 1H NMR experiments and molecular dynamics (MD) simulation were used to analyze the peptidyl-prolyl cis/trans imide bond isomerization. The data indicated that the side-chain O-phosphorylation of the Xaa residues preceding proline affected evidently the isomerization and thereby regulated the peptides conformations. The charges of the phosphate moiety as well as their steric effects might be the driving force for the conformational changes of these phosphopeptides.Moreover, the obtained most stable multiple configurations and their statistic cis/trans concentration distribution in MD simulation were basically consistent with the NMR experiments, which demonstrated that phosphorylation increased the cis conformation of the peptide and the maximum cis ratio is given while the phosphate group has no negative charge.

Objective:To observe the effect of herbal-partitioned moxibustion (HPM) on pain-related behavior and emotion in a rat model of chronic inflammatory visceral pain, and to investigate the mechanism.
Methods:Twenty-four male Sprague-Dawley (SD) rats were randomly divided into three groups:a normal group, a model group and an HPM group. Except for the normal group, rats in the other two groups were clystered with mixed liquor of Trinitrobenzene Sulfonic Acid (TNBS) and 50%ethanol to induce the chronic inflammatory visceral pain model. After the models were established successfully, rats in the HPM group were treated with HPM at bilateral Tianshu (ST 25) and Qihai (CV 6). Rats in the normal group and the model group were only fixed as those in the HPM group without treatment. Abdominal withdrawal reflex (AWR) score, mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were adopted to detect the visceral and somatic pain;meanwhile, open field test (OFT) and elevated plus maze test (EPMT) were employed to evaluate pain emotions such as depression and anxiety.
Results:Compared with the normal group, AWR scores of the model group were significantly increased under different stimulus expansion pressure level (P<0.01), MWT and TWL were significantly decreased (P<0.05);in OFT, the values of horizontal activities and vertical activities were significantly decreased (P<0.01);in EPMT, the proportion of the number of entry into the open arms (OE%) and that of residence time in the open arms (OT%) significantly decreased (P<0.01), indicating that the model was successful. Compared with the model group, the AWR score of the HPM group was decreased significantly (P<0.05), MWT and TWL were significantly increased (P<0.05), the values of horizontal activities and vertical activities in the model group were significantly increased (P<0.01);in OFT and EPMT, OE%and OT%were significantly increased (P<0.01).
Conclusion:HPM has analgesic effect on chronic inflammatory visceral pain. It can reduce the visceral and somatic pain in rats and markedly improve the emotions such as anxiety and depression induced by chronic visceral pain.

ObjectiveTo investigate the immunity and seroprevalence of hepatitis A and to identify the risk factors of hepatitis A infection in 0-18 year-old children and adolescents in Shanghai.MethodsSubjects were enrolled by stratifying and clustering random sampling method.Questionnaire interview was applied to investigate the socio-demographic and behavioral factors related to hepatitis A virus (HAV),and information on HAV immunization was abstracted from the immunization registration book of each subject.The enzyme-linked immunosorbent assay (ELISA) was used to qualitatively detect HAV IgM and quantitatively measure total HAV antibody in all subjects.Risk factors associated with HAV among the subjects without HAV vaccination were analyzed.ResultsA total of 2431 subjects were enrolled in the present study with negative HAV IgM antibody and total HAV antibody in 1483 subjects were sero-positive with positivity rate of 61％.Total HAV antibody positivity rates were declined with age increasing and were significantly higher in subjects with HAV vaccination than those without HAV vaccination records.Salad food,eating together without food separation in school and endoscopy inspection were risk factors for HAV infection.ConclusionsHAV vaccination strategies remarkably improve the total HAV antibody seropositive rate in children and adolescents in Shanghai.The risk of HAV infection exists if HAV vaccination is not administrated comprehensively.Therefore,strengthening HAV vaccination and health education are important for children and adolescents to prevent and control of hepatitis A in Shanghai.

Objective: To explore the effect of group cognitive behavioral therapy (GCBT) on cognitive control among college students with high obsessive compulsive traits, to provide basic information for the psychological counseling intervention for college students. Methods: From March to April 2019, 687 students were conveniently selected from 2 universities in Hefei. According to the inclusion and exclusion criteria, 58 students with high obsessive traits were selected and divided into experimental group ( n =29) and control group ( n =29) by random number table method. The experimental group received cognitive behavioral group counseling for 4 weeks (1.5 h each time, twice a week), while the control group receive no intervention. The Obsessive Compulsive Inventory Revised (OCI-R), Stroop Color Word Test (SCWT), Digital Span Test (DST), and Wisconsin Card Sorting Test (WSCT) were used to assess in two groups at baseline and 4 weeks later. Results: After 4 weeks, the scores of OCI-R in the GCBT group (10.28±7.22) was lower than that of in the control group (15.90±10.20) ( t=2.42, P<0.05). Before and after intervention, compared with the control group [(21.89±6.63, 20.52±7.37)s, (8.62±4.43, 8.04±4.84)s] in Stroop C and Stroop interfere effects (SIE), the GCBT group [(22.14±4.92, 16.81±3.43)s, (8.36±3.87, 4.82±1.86)s], the interaction of time group was statistically significant ( F =14.60, 10.54, P <0.05). Compared with the control group (6.21±1.35, 6.55±1.45)times, the scores of DST reverse in the GCBT group (6.31±1.44, 7.24±1.38) times were statistically significant ( F=3.96, P <0.05). Conclusion It suggests that cognitive behavioral group counseling can improve the inhibitory control and working memory of college students with high obsessive compulsive traits, but does not change the cognitive flexibility.

OBJECTIVETo investigate the correlation between tissue factor (TF) expression and hepatic metastasis and prognosis in rectal cancer.

METHODSTF expression was retrospectively studied by immunohistochemical method in specimens of 40 rectal cancer, 3 hepatic metastasis and 6 benign adenoma with relation to their clinicopathologic data.

RESULTS1. TF expression was detected in 20 (50%) of the 40 primary rectal cancer specimens and all the 3 hepatic metastatic specimens, but not in the 6 benign adenoma or normal mucosa of rectum, 2. Significant correlation was observed between TF expression and synchronic hepatic metastasis (P = 0.002) and heterochronic hepatic metastasis (P = 0.001) and 3. TF was a risk factor for the prognosis of primary rectal cancer (P = 0.024).

CONCLUSIONTissue factor expression may play a role in the process of developing hepatic metastasis. It may be considered as a new clinical indicator for monitor of hepatic metastasis and prognosis of primary rectal cancer.

Adenoma ; metabolism ; pathology ; Humans ; Immunohistochemistry ; methods ; Liver Neoplasms ; metabolism ; secondary ; Logistic Models ; Predictive Value of Tests ; Proportional Hazards Models ; Rectal Neoplasms ; metabolism ; pathology ; Rectum ; metabolism ; Retrospective Studies ; Staining and Labeling ; methods ; Thromboplastin ; biosynthesis

Objective:To evaluate the clinical efficacy and safety of fractional CO 2 laser for the treatment of vaginal relaxation syndrome. Methods:Twenty-one females with vaginal relaxation syndrome receiced between December 2016 and January 2017 in our department were treated for 3 times at 1 month interval with femilift fractional CO 2 laser. Vaginal palpation, vaginal squeeze pressure test, vaginal ultrasound, pelvic floor MRI, and Incontinence Questionnaire Short-Form (ICIQ-SF) assessments were performed at baseline and at post-treatment. Results:Significant improvement in female sexual dysfunction and stress urinary incontinence was especially seen based on patient＇s feelings after treatments. A noted progressive differences were observed in FSFI and vaginal pressure test, which were much higher after treatment than before treatment ( P<0.01). Moreover, in 13 cases suffering from mild to moderate stress urinary incontinence (SUI), fractional CO 2 laser treatment was associated with a significant improvement of ICIQ-SF scores ( P<0.01). All subjects successfully completed the study with no adverse events such as infections, bleeding and pain, etc. Conclusions:Fractional CO 2 laser treatment is an effective treatment for female sexual dysfunction and stress urinary incontinence caused by vaginal relaxation syndrome.

Objective:To study the difference in the extraction efficiency of the novel coronavirus (2019-nCoV) nucleic acid by using magnetic beads method, centrifugal column method and one-step method.Methods:On March 5, 2021, 10 throat swabs were collected from the staff working in the nucleic acid sampling room in Department of Clinical Laboratory, Affiliated Taikang Xianlin Drum Tower Hospital, Medical School of Nanjing University. The positive quality control samples were mixed into the swabs and used as mock positive samples. The RNA was extracted from simulated positive samples and their diluted samples by using magnetic beads method, centrifugation column method and one-step method. The purity ( A260/ A280 ratio) and concentration of the nucleic acid obtained were measured by micro-uv photometry, and fluorescence quantitative PCR was performed to compare the CT value and extraction efficiency. The three methods were used to extract the simulated weak positive specimens and to compare the difference of CT values after amplification. The measurement data that followed normal distribution were expressed by xˉ±s, the t test was used for comparing in the same group, and single factor analysis of variance was used for comparing among multiple groups. A P value smaller than 0.05 indicated a significant difference. Results:2019-nCoV nucleic acid extracted by magnetic bead method, centrifugal column method and one-step method could amplify positive results. There was no significant difference between the CT value of RNA amplification extracted by magnetic bead method and one-step method ( t=? 0.995 , P=0.376). The CT values of orf1ab gene amplified by centrifugal column method, magnetic bead method and one-step method were 29.28±0.06, 30.82±0.14 and 29.79±0.01 respectively ( F=11.196 , P=0.041). The CT values of E gene were 28.52±0.40, 27.33±0.78 and 27.38±0.13 respectively ( F=3.407, P=0.169). The CT values of N gene were 28.61±1.02, 27.24±0.20 and 27.25±0.47, respectively ( F=2.880 , P=0.020). The CT values of human genes extracted by centrifugal column method, magnetic bead method and one-step method were 19.68±0.36, 20.14±0.06 and 20.58±0.49 respectively, which was statistically significant ( F=4.904, P=0.048). The CT value of amplified human gene was affected by the dilution of human samples twice. The CT value of undiluted samples was smaller than that of diluted samples twice, with a difference of 2.95±0.22, which was statistically significant ( t=?3.025, P=0.039). The extraction time of one-step method, magnetic bead method and centrifugal column method were (15.00±1.50), (20.00±1.50) and (40.00±5.5) min respectively, and the difference was statistically significant ( F=688 , P=0.027). Conclusions:Magnetic bead method, centrifugal column method and one-step method can be used to extract 2019-nCoV nucleic acid, for the centrifugal column method has a higher extraction efficiency than the magnetic bead method and the one-step method. The one-step method is the fastest, followed by the magnetic bead method and the centrifugal column method. A large number of clinical samples can be processed using the magnetic bead method and one-step method. One-step rapid nucleic acid test can also be performed on samples from emergency and fever clinics. It is not recommended to dilute specimens for testing. In order to improve the detection rate, extracting RNA from highly suspected samples with negative initial nucleic acid test by centrifugal column method is suggested.

Objective:To construct and verify the nomogram prediction model based on inflammatory indicators, underlying diseases, etiology and the British Thoracic Society modified pneumonia score (CURB-65 score) in adults with severe community acquired pneumonia (CAP).Methods:The clinical data of 172 adult inpatients first diagnosed as CAP at Taikang Xianlin Drum Tower Hospital from January 2018 to December 2021 were divided into severe and non-severe diseases groups according to the severity of their conditions. The baseline conditions (including gender, age, past history, comorbidities and family history), clinical data (including chief symptoms, onset time, CURB-65 score), first laboratory results on admission (including whole blood cell count, liver and kidney function, blood biochemistry, coagulation function, microbiological culture results) and whether the antimicrobial therapy was adjusted according to the microbiological culture results were recorded in both groups. Univariate analysis was used to screen for differential indicators between severe and non-severe patients. After covariate analysis, multi-factor Logistic regression analysis was performed based on the Aakaike information criterion (AIC) forward stepwise regression method to rigorously search for risk factors for constructing the model. Based on the results of the multi-factor analysis, a nomogram prediction model was constructed, and the discriminatory degree and calibration degree of the model were assessed using the receiver operator characteristic curve (ROC curve) and calibration curve.Results:A total of 172 adult CAP patients were included, 48 in severe group and 124 in non-severe group. The median age was 74 (57, 83) years old, onset time was 5.0 (3.0, 10.0) days, total number of comorbidities was 3 (2, 5), including 58 cases (33.7%) with hypertension and 17 (9.9%) with heart failure, 113 (65.7%) with CURB-65 score≤1, 34 cases (19.8%) had a CURB-65 score = 2 and 25 cases (14.5%) had a CURB-65 score≥3. Univariate analysis showed that there were statistically significant differences between the two groups in age, smoking history, CURB-65 score, heart rate, onset time, total comorbidity, pathogenic microorganisms, fibrinogen (FIB), D-dimer, C-reactive protein (CRP), procalcitonin (PCT), platelet-to-lymphocyte ratio (PLR), neutrophil-to-lymphocyte ratio (NLR), and alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Multi-factor Logistic regression analysis showed that hypertension [odds ratio ( OR) = 3.749, 95% confidence interval (95% CI) 1.411 to 9.962], heart failure ( OR = 4.616, 95% CI was 1.116 to 19.093), co-infection ( OR = 2.886, 95% CI was 1.073 to 7.760), history of smoking ( OR = 8.268, 95% CI was 2.314 to 29.537), moderate to high CURB-65 score ( OR = 4.833, 95% CI was 1.892 to 12.346), CRP ( OR = 1.012, 95% CI was 1.002 to 1.022), AST ( OR = 1.015, 95% CI was 1.001 to 1.030) were risk factors for severe CAP (all P < 0.05). The filtered indicators were included in the nomogram model, and the results showed that the area under the ROC curve (AUC) for the model to identify patients with severe adult CAP was 0.896, 95% CI was 0.840 to 0.937 ( P < 0.05), and the calibration curve showed that the predicted probability of severe CAP was in good agreement with the observed probability (Hosmer-Lemeshow test: χ2 = 6.088, P = 0.665). Conclusions:The nomogram model has a good ability to identify patients with severe adult CAP and can be used as a comprehensive and reliable clinical diagnostic tool to provide a evidence for timely intervention in the treatment of adults with severe CAP.