Objective : To evaluate the excess mortality risk related to heat wave in Ningbo, Zhejiang from 2013 to 2018, so as to provide a basis for formulating coping strategies for heat wave. Methods : The data of daily mortality, meteorological and air quality from May to October in Ningbo from 2013 to 2018 were obtained from Ningbo Center for Disease Control and Prevention, Ningbo Meteorological Bureau and Environmental Monitoring Center of Ningbo, respectively. The generalized linear model ( GLM ) and distributed lag non-linear model ( DLNM ) were used to estimate the associations between heat wave and cause-specific mortality. Results : Among 1 104 days of the study period, 18 heat waves occured and lasted for 132 days, accounting for 11.96%. A total of 102 954 deaths were reported in the same period. The risks of mortality in circulatory system diseases ( RR=1.09, 95%CI: 1.03-1.16 ), respiratory system diseases ( RR=1.14, 95%CI: 1.04-1.25 ), digestive system diseases ( RR=1.38, 95%CI: 1.15-1.65 ), nervous system diseases ( RR=1.32, 95%CI: 1.08-1.61 ), mental disorders ( RR=1.51, 95%CI: 1.12-2.03 ) and accidental injury ( RR=1.18, 95%CI: 1.06-1.32 ) and all causes ( RR=1.10, 95%CI: 1.06-1.14 ) increased at lag 0-1 day of heat wave. The total excess death related to heat wave was 1 218 ( 95%CI: 731-1 705 ) . The excess deaths of circulatory system diseases, respiratory system diseases, accidental injury, digestive system diseases, nervous system diseases, mental disorders, urinary system diseases and endocrine system diseases were 313 ( 95%CI: 104-556 ), 206 ( 95%CI: 59-368 ), 164 ( 95%CI: 55-292 ), 122 ( 95%CI: 48-208 ), 69 ( 95%CI: 17-131 ), 56 ( 95%CI: 13-113 ), 18 ( 95%CI: -15-64 ) and 3 ( 95%CI: -51-72 ). The excess deaths of urinary system and endocrine system diseases was not statistically significant ( P>0.05 ). Conclusion Heat wave can increase the mortality risk on the day and after a day in Ningbo from 2013 to 2018. Circulatory system diseases, respiratory system diseases and accidental injury rank top three in excess deaths.

Objective: To identify the most appropriate meteorological variable for forecasting the health risk of high temperatures. Methods: The surveillance on causes of death, meteorological data and surveillance on air quality among registered residents in Ningbo City, Zhejiang Province during the period between May and October from 2013 to 2019 were collected. The association models of daily minimum temperature, average daily temperature, daily maximum temperature, daily minimum heat index, average daily heat index, daily maximum heat index, average daily apparent temperature and torridity index with deaths and years of life lost (YLL) were created using time series analysis and distributed lag non-linear models, and the model fitting effect was evaluated using the minimum Akaike information criterion (AIC) procedure. The most appropriate meteorological variable for forecasting gender-, age- and mortality-specific health risks of high temperatures was identified. Results: A total of 120 628 deaths were reported during the study period, with daily deaths of 94 cases, and daily YLL rate of 19.74 person-years/105. Except for daily minimum heat index and torridity index, the exposure-response relationships between other six meteorological variables and deaths and overall YLL rate all appeared a “J” shape. The lowest AIC values and the optimal model fitting effects were measured for the association models between average daily temperature and whole populations, females, subjects at ages of 65 years and older, and deaths and YLL rates due to circulatory diseases and respiratory diseases. Conclusion High model fitting effects are observed between average daily temperature and deaths and YLL rates, which are more suitable for forecasting the health risk of high temperature.

ObjectiveTo study the expression of microRNA-21 ( miR-21 )in serum of patient with diffuse large B cell lymphoma (DLBCL) and DLBCL cell lines and validate the significance of miR-21 in early diagnosis,genotyping and prognosis estimates of DLBCL.MethodsmiR-21 expression were detected by fluorescent quantity polymerase chain reaction (FQ-PCR)in 9 lymphoma cell lines (OCI-Ly1,OCI-Ly3,OCI-Ly4,OCI-Ly7,OCI-Ly8,OCI-Ly10,OCI-Ly18,OCI-Ly19 and HBL),the serum from DLBCL patients (n =62) and health controls (n =50 ).Kaplan-Meier survival analysis was carried out during the relapsefree survival period of DLBCL patients to explore the relationship between the prognosis and microRNA expression level.ResultsReal time FQ-PCR result indicated that miR-21 expression was higher in DLBCL cell lines than that in normal B cells (BC).miR-21 expression in normal B cell and 9 DLBCL cell lines separately were 1.04 ± 0.02,2.30 ± 0.35,237.97 ± 56.19,5.27 ± 0.83,3.40 ± 0.30,11.22 ± 2.70,133.55 ± 16.78,6.63 ±0.24,4.91 ±0.37 and 81.59 ±6.64.Compared with BC,the expression of miR-21 were higher in all 9 DLBCL cell lines ( t =7.3,13.7,21.0,6.2,8.8,13.6,6.5,39.5,18.1 ;P ＜ 0.01 ).miR-21 expression segregates with specific molecular subgroups of DLBCL The expression was higher in the ABC type cell lines (OCI-Ly3,OCI-Ly10,HBL) than GCB type cell lines (OCI-Ly1,OCI-Ly4,OCI-Ly7,OCI-Ly8,OCI-Ly18,OCI-Ly19;t =11.18,P ＜ 0.01 ).Consistent with the cell line models,miR-21 expression levels were higher in serum from DLBCL patients [21.38 (10.26-45.21 )] than from controls [1.87 ( 1.05-3.97 ),U =168,P =0.000],and the levels were higher in DLBCL cases with an ABC-type [28.68 ( 14.92-98.44 )] than those in GCB-type [18.30 ( 7.32-33.46 ),U =336,P =0.043].MiR-21 expression levels were different in sera from different clinical stage DLBCL patients.The miR-21 level in serum of patients with subgroup ABC and subgroup GCB in stage Ⅰ and Ⅱ were 47.49( 25.65-295.41 ) and 24.74( 16.08-50.38) respectively and in stage Ⅲ and Ⅳ were 16.66 ( 5.35-44.30 ) and 11.96 ( 4.10-21.05) respectively.The levels were higher in DLBCL cases withⅠ -Ⅱ stage than those with Ⅲ-Ⅳ stage (U =62,P =0.013 in GCB type; U =53,P =0.014 in ABC type).Moreover,compare with relapse-free survival in DLBCL patients,high miR-21 expression was associated with well prognosis ( U =259,P =0.035).ConclusionsMiR-21 is high expression in DLBCL cell lines and DLBCL patients serum.miR-21 level in sera from DLBCL patients is associated with clinical stage,molecular subgroup and prognosis estimates.MiR-21 may serve as a new biomarker to early detection,genotyping and prognosis estimates of DLBCL.

Objective To study the infection rate of fusobacterium nucleatum cancer re appeared in patients with colorectal cancer before and after radiotherapy,and the changes after cancer recarrence.Methods A total of 20 persons receiving physical examination were recruited in the control group and collected the stool specimens,and 40 colorectal cancer patients were selected in the study group.All of the subjects in the study group were collected stool specimens before operation 3 days and after operation 5 day,after radiation therapy 7 days and 30 days.The patients were followed-up 1 year.The bacterial fluid was collected by filtration,and real-time fluorescence quantitative PCR was used to detect the expression of fusobacterium nucleatum gene in feces.Results The positive rate of fecal fusobacterium fusiformis was 30% in the study group and 5% in the control group.The gene relative expression of 12 colorectal cancer patients before operation 3 days and after operation 5 days,after radiation therapy 7 days and 30 days were 5.20±0.34,8.50±0.45,1.20±0.22,0.20±0.15.The fusobacterium nucleatum gene expression of 12 patients with positive fusobacterium after operation 5 days was significantly increased compared with that before operation 3 days(t=10.419,P=0.001),which after radiation therapy 7 days and 30 days was significant lower than that before operation 3 days(t=12.728,P=0.001;t=25.889,P=0.001).Six patients recurred among 1 year,the fusobacterium nucleatum gene expression was 7.2±0.56,which was significant higher than that after radiation therapy 7 days.Conclusion The infection of fusobacterium nucleatum might be a risk factor for colorectal cancer,and the gene relative expression might be an early warning indicator of recurrence.

Objective To predict and identify the linear B-cell epitopes in the major group 3 allergen derived from Derma-tophagoides farina(Der f 3). Methods The linear B-cell epitopes of Der f 3 allergen were analyzed based on the physicochemi-cal properties of amino acids including antigenicity,surface accessibility,flexibility,hydrophilicity,beta-turn by online bioinfor-matics softwares. The eight predicted peptides of linear B-cell epitopes were artificially synthesized and incubated with three aller-gic serum pools(4 serum samples in each),which were consisted of total 12 serum samples from the allergic individuals,and the strong positive epitopes were selected. Results Eight B-cell epitopes from Der f 3 were predicted successfully. Five of eight B-cell epitopes were identified with strong IgE-binding abilities followed by specific IgE assay. The amino acid sequences of them were following:33KAKAGDCP40, 86HASGGEKIQVAEIYQHENYDSMTID110, 118LKTPMTLDQTNAKPVPLPPQGSDVKVG144, 156QEGSYSLP163 and 199DVANGGVDSCQGDSGGPVVD218. Conclusions Five linear B-cell epitopes of Der f 3 allergen have been identified successfully. This result might provide a basis of the diagnosis and treatment for asthma.

Objective:To explore therapeutic effect of Fasudil combined Salmeterol Xinafoate and fluticasone propio‐nate powder for inhalation (Seretide) on patients with chronic obstructive pulmonary disease (COPD) combined pul‐monary arterial hypertension (PAH ) .Methods :A total of 120 patients ,who hospitalized in our department from Jan 2013 to Oct 2014 and conformed to diagnostic standards of COPD and PAH ,were selected .According to ran‐dom number table ,they were equally divided into routine treatment group (received routine therapeutic measures ) , Fasudil group (received Fasudil based on routine treatment group ) and combined treatment group (received Fasudil combined Seretide based on routine treatment ) . Pulmonary function indexes , mean pulmonary arterial pressure (mPAP) ,pulmonary arterial systolic pressure (PASP) ,6min walking distance (6MWD) and blood gas indexes were observed and compared among three groups before and after treatment .Results:Compared with routine treatment group after treatment ,there were significant reductions in mPAP [(54.1 ± 10.3) mmHg vs .(51.3 ± 9.5) mmHg vs . (48.5 ± 10.5) mmHg] and PASP [ (72.4 ± 9.7) mmHg vs .(63.4 ± 9.3) mmHg vs .(61.6 ± 9.1) mmHg] ,and sig‐nificant rise in 6MWD [ (259.4 ± 37.0) m vs .(274.2 ± 36.5) m vs .(288.3 ± 47.5) m] ,forced expiratory volume in one second [FEV1 ,(1.44 ± 0.32) L vs .(1.59 ± 0.38) L vs .(1.87 ± 0.34) L] and FEV1/forced vital capacity (FVC) [ (47.2 ± 11.9)% vs .(50.3 ± 12.1)% vs .(54.6 ± 11.7)% ];significant rise in partial pressure of oxygen in artery [PO2 ,(64.3 ± 9.8) mmHg vs .(68.9 ± 8.2) mmHg vs .(76.9 ± 9.5) mmHg] and saturation of arterial blood oxygen [SaO2 ,(65.0 ± 8.2)% vs .(71.0 ± 9.8)% vs .(76.8 ± 9.4)% ] ,and significant reduction in partial pressure of carbon dioxide in artery [PCO2 ,(63.6 ± 9.5) mmHg vs .(58.5 ± 9.6) mmHg vs .(51.3 ± 7.9) mmHg] in Fasud‐il group and combined treatment group ,and those of combined treatment group were significantly improved com‐pared to those of Fasudil group , P<0.05 or <0.01. Actual base excess of combined treatment group was signifi‐cantly higher than the other two groups , P<0. 01 both . Conclusion:Fasudil combined Seretide can significantly im‐prove pulmonary function reduction ,improve PAH ,quality of life and prognosis in COPD + PAH patients .

Full-length coxsackie adenovirus receptor (CAR) eukaryotic expression plasmid was transfected into an ovarian cell line, SKOV3, and its effect on the change of malignant metastasis phenotype was explored. CAR mRNA and protein expression levels among 4 ovarian cancer cell lines (A2780, SKOV3, SW626, CAOV3) and the positive control 293 (a transformed human embryo kidney cell line) was detected by using semi-quantitative RT-RCR and Western blot and compared. CAR-negative SKOV3 was transfected with the eukaryotic expression plasmid containing a full-length CAR cDNA and mock-vector respectively. The positive clones were screened by G418. The biological behavior changes of positive transfected cells were gauged by colony formation in soft agar assay and cell adhesion assay. Among the cell lines, there were obviously different CAR expression levels. CAR could not be detected in SKOV3. In transfected cell group, CAR expression was enhanced obviously as compared with non-transfected or mock-transfected groups. Cell adhesion in the transfected group was promoted. The number of colony formation was reduced significantly in transfected groups (25.32 +/- 8.91) as compared with that in non-transfected group (88.75 +/- 13. 98) and mock-transfected group (82.53 +/- 19.37). Among the 4 ovarian cancer cell lines, CAR expression level was variable. Exogenous CAR expression had a potential role in inhibiting the malignant metastasis phenotype of ovary cancer cells.
Cell Line, Tumor ; Lymphatic Metastasis ; Neoplasm Invasiveness ; Ovarian Neoplasms/metabolism ; Ovarian Neoplasms/*pathology ; Phenotype ; Receptors, Virus/*biosynthesis ; Receptors, Virus/genetics ; Transfection

Objective: To explore the relationship between different temperature indicators and hospital admission for childhood pneumonia. Methods: The hospital admissions for pneumonia in children aged 0-14 years and meteorological data in Ningbo from 2015 to 2017 were collected. A distributed lag non-linear model combined with a generalized linear model was employed to analyze the exposure-response relationships between different temperature indicators（daily average,minimum and maximum temperature;the first percentile as low temperature and the 99th percentile as high temperature）and hospital admission for childhood pneumonia. Results: A total of 4 542 cases of childhood pneumonia were recruited. There were obvious seasonal fluctuations found in the inpatient volume of childhood pneumonia,which peaked in winter and bottomed in summer. After adjusting for potential confounding variables such as relative humidity,PM2.5,long term trend and seasonal trend,the results suggested that after exposed to whether low or high temperature,the inpatient volume of childhood pneumonia would increase. When the daily average temperature and daily minimum temperature were employed,the effect of high temperature on the increase of inpatient volume for childhood pneumonia was statistically significant and the cumulative relative risk for a lag of 0-7 days were 1.52（95%CI：1.04-2.23）and 1.59（95%CI：1.08-2.34）,respectively. When the daily maximum temperature was employed,the effect of low temperature on the increase of inpatient volume for childhood pneumonia was statistically significant and the cumulative relative risk for a lag of 0-7 days were 1.30（95%CI：1.02-1.66）. Conclusion Our findings suggested that an increased risk of hospital admission for childhood pneumonia was associated with both low and high temperature.

OBJECTIVE:To systematically evaluate the effectiveness and safety of escitalopram and duloxetine in the treatment of depression, and provide evidence-based reference for clinical treatment. METHODS:Retrieved from PubMed, Wanfang database,VIP,CNKI and CBM,randomized controlled trials (RCTs) about escitalopram (trial group) and duloxetine (control group) in the treatment of depression were collected. Meta-analysis was conducted by using Rev Man 5.3 software after data extraction and quality evaluation according to bias risk assessment tool recommended by system evaluator manual 5.3. RESULTS:Finally 25 RCTs were included,involving 2621 patients. The results of Meta-analysis showed that there was no statistical significance in total response rate between 2 groups after 1,2,4,6,8 weeks of treatment or cure rate between 2 groupsafter 4,6,8 weeks of treatment (P>0.05). There was no statistical significance in total response rate [RR=0.96,95％CI(0.88, 1.05),P=0.42] or cure rate [RR=0.91,95％CI(0.78,1.06),P=0.24] of female patients,as well as total response rate [RR=0.96, 95％CI(0.84,1.11),P=0.61] or cure rate [RR=0.90,95％CI(0.54,1.49),P=0.69] of elderly patients between 2 groups. The incidence of constipation [RR=0.59,95％CI (0.42,0.81),P=0.001],dry mouth [RR=0.65,95％CI(0.51,0.82),P=0.0004], nausea [RR=0.68,95％CI(0.56,0.83),P=0.0002] and decreased appetite [RR=0.74,95％CI(0.55,0.99),P=0.04] in trial group were significantly lower than control group,with statistical significance. CONCLUSIONS:The effectiveness of escitalopram is similar to duloxetine in the treatment of depression,but the safety of escitalopram is better than duloxetine.

OBJECTIVETo construct a vector encoding T-cell epitopes of major allergen group 1 of Dermatophagoides pteronyssinus as a vaccine delivered by MHC class II pathway.

METHODSThe nucleotide sequences of the 3 target genes were synthesized, including TAT, IhC and the recombinant fragment of Der p 1 encoding 3 T-cell epitopes. After amplification of the 3 target fragments by PCR and digestion with corresponding restriction endonucleases, the recombinant gene TAT-IhC-Der p 1-3T was ligated using T4 DNA ligase and inserted into the prokaryotic expression vector pET28a(+) to construct the recombinant plasmid pET-28a(+)-TAT-IhC-Der p 1-3T, which was confirmed by digestion with restriction endonucleases and sequencing. The recombinant vector was transformed into E. coli strain BL21 (DE3) and induced with IPTG, and the induced protein TAT-IhC-Der p 1-3T was detected by SDS-PAGE. After purification, the recombinant protein was confirmed by Western blotting and its allergenicity tested using IgE-binding assay.

RESULTSThe recombinant plasmid pET-28a-TAT-IhC-Der p 1-3T was successfully constructed as confirmed by restriction endonuclease digestion and sequencing and the expression of the recombinant protein TAT-IhC-Der p 1-3T was induced in E. coli. Western blotting verified successfull purification of the target protein, which showed a stronger IgE-binding ability than Der p 1.

CONCLUSIONWe successfully constructed a recombinant expression vector pET-28a-TAT-IhC-Der p 1-3T expressing a T-cell epitope vaccine delivered by MHC II pathway with strong IgE-binding ability, which provides a basis for further study on specific immunotherapy via MHC class II pathway.

Allergens ; immunology ; Animals ; Antigens, Dermatophagoides ; immunology ; Arthropod Proteins ; immunology ; Base Sequence ; Cloning, Molecular ; Cysteine Endopeptidases ; immunology ; Dermatophagoides pteronyssinus ; Epitopes, T-Lymphocyte ; Escherichia coli ; Gene Expression ; Genes, MHC Class II ; Genetic Vectors ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; immunology ; Vaccines ; immunology