Objective To construct a plasmid DNA encoding G glycoprotein of respiratory syncytial virus(RSV) and investigate the protective immune response against RSV infection. Methods Recombinant plasmid DNA of pcDNA3.1~G was constructed by standard RT-PCR based cloning procedure. The immunogenicity of recombinant G protein transiently expressed in HEK293 cells was detected by Western blot. BABL/c mice were intramuscularly immunized with pcDNA3.1~G. Samples of lung, sera, bronchoalveolar lavage fluid(BALF) were collected before and after RSV challenge; virus titer in lung was detected by viral titration; sections of paraffin embedding lung tissues were stained by haematoxylin and eosin(HE) for histological analyses; sera anti-RSV IgG levels were examined by ELISA; Th1/Th2 cytokine were detected by ELISA kit, the T lymphocyte subsets of BALF was determined by immunefluorescence staining followed by flow cytometry. Results Plasmid DNA of pcDNA3.1~G was successfully constructed. The expressed target protein possesses immunogenicity. After challenge, pcDNA3.1~G immunized mice presented relieved pathological changes in lung as well as reduced lung viral titers. The RSV specific IgG was detected in sera of immunized mice. There was significantly increased number of CD25~+CD4~+ T cells in mice BALF. Conclusion We constructed a pcDNA3.1~G plasmid DNA vaccination which can induce evident protective cellular immunity against RSV infection in mice with the increased number of CD25~+CD4~+ T cell subpopulation.

Objective To investigate the problems existing in hospital transfusion records,analyze unreasonable factors,and improve the quality of blood transfusion.Methods Investigation of our hospital from January 2016 to January 2017 with blood transfusion medical records,according to hospitalization number,900 cases were selected,using random number table.Investigating blood transfusion records on each record integrity,according toGuidelines for Surgical and Traumatic Blood Transfusion and Internal medicine transfusion guide in clinical blood transfusion technical specifications.The common blood components such as red blood cells,plasma and cryoprecipitate infusion and rationality evaluation.Results There were 583 surgical departments and 557 non-surgical departments.There was a significant difference between the surgical department and the non-surgical department.The reasonable rate of non-operation department was higher than that of the operation department (x2 =7.723,P=0.021).The rational rate of the department was 93.8％,while the operation department was only 88.0％;900 blood transfusion records of four kinds of blood components of the rationality of the difference was statistically significant (x2=214.767,P＜0.001).Non-surgical department of erythrocyte,plasma,cold precipitate reasonable rate than the surgical department;900 medical records in 55 records failed,mainly after the assessment of incomplete blood transfusion,no recorded blood transfusion reaction.There were significant differences in the failure rate between the surgical department and the non-surgical department (x2 =4.613,P=0.032).Conclusion Some physicians transfusion indications,for evaluation before and after blood transfusion blood insufficient attention to curative effect evaluation,blood transfusion is not reasonable,and in the operation department,do not take the blood transfusion medical record writing,hospitals should strengthen the blood transfusion blood transfusion continuously improve the scientific and normative management.

Objective To determine the effects of high-frequency jet ventilation (HFJV) on oxidative stress in patients during one-lung ventilation (OLV). Methods Forty-five patients undergoing elective radical esophageal cancer resection were divided into three groups with 15 cases each by random digits table: two lung ventilation group ( group A), OLV group (group B), HFJV- OLV group ( group C, working pressure 1 kg/cm2 and frequency 100 times/min). Venous blood samples were taken before induction (T0),at30min (T1),90min (T2),150min (T3) after OLV and the end of operation (T4) for measuring serum superoxide dismutase(SOD),malondialdehyde (MDA) and nitric oxide (NO). Results SOD was lower at T2-T4 in group B[(47 ± 10), (37 ±9), (41 ±7) kU/L] and group C[(58 ± 12), (51 ± 11), (49 ± 9) kU/L] than those in group A [(78 ±8), (75 ±7), (79 ±6) kU/L](P＜ 0.05),and MDA and NO were lower at T1-T4 in group B and group C than that in group A(P＜ 0.05). SOD was higher at T3 in group C than that in group B (P ＜ 0.05), MDA at T1-T4 and NO at T2-T4 were lower in group C than those in group B (P ＜ 0.05). Conclusion HFJV can effectively decrease oxidative stress in patients during OLV.

Several aquatic species and their enviroments were examined for presence of Vibrio parahaemo-lyticus between 2007 and 2008 in the coastal areas in Zhejiang province, and some virulence-related genes such as tdh, trh, ureC and vscC2 were investigated from the isolates. V. parahaemolyticus was recovered from 70% of the samples tested (395/566). The genes tdh, trh and ureC existed in 10.1%, 20.0% and 11.1% respectively from 395 isolates. Among the 40 tdh-positive isolates, 32.5% harbored the vscC2 gene, one of the type three secretion system 2 (T3SS2) gene family. Thirty-eight of the 40 tdh-positive isolates were positive for the Kanagawa phenomenon. Out of 44 trh-and-ureC-positive isolates, only six exhibited urease phenotype. Overall, this study reveals the significant prevalence of Vibrio parahaemolyticus in seafoods and their habitats with high diversity of virulence genes. Representative V. parahaemolyticus isolates could beused for further investigation into their pathongenecity, functional genomics, and molecular evolution.

We investigated the roles of the crude antigen(CA) of Clonorchis sinensis and excretory secretory products (ESPs) in the polarization of Th1 and Th2 cells.Bone marrow-derived cells were generated from BALB/c mice and isolated into immature DCs;immature DCs were then treated with either CA (CA stimulated group),ESPs (ESPs stimulated group),LPS (positive control group) or PBS (negative control group) for 24 hours.Then the CD4+T cells were isolated from mouse spleen by using anti mouse-CD4 Microbeads,and further cocultured with stimulated DCs for another 72 hours.The purities of DCs and CD4+ T cells were evaluated by flow cytometry and the expressing levels of T-bet mRNA and GATA-3 mRNA were detected by real-time PCR.ELISA was used to detect the levels of IFN-γ and IL-4 cytokines in the supernatant.mRNA levels of T-bet and GATA-3 in the ESPs group were higher than those in PBS-stimulated group (P＜0.05).The concentrations of IFN-γ and IL-4 cytokines in the culture were increased in the ESPs group,compared with PBS stimulated group(P＜0.05).IFN-γ but not IL-4 was increased in CA group (P＜0.05).The results implied that CA might play a role in Th1 type immune response,and ESPs likely play roles in both Th1 and Th2 immune responses.

BACKGROUND/OBJECTIVES: D. candidum is a traditional Chinese food or medicine widely used in Asia. There has been little research into the anticancer effects of D. candidum, particularly the effects in colon cancer cells. The aim of this study was to investigate the anticancer effects of D. candidum in vitro and in vivo. MATERIALS/METHODS: The in vitro anti-cancer effects on HCT-116 colon cancer cells and in vivo anti-metastatic effects of DCME (Dendrobium canidum methanolic extract) were examined using the experimental methods of MTT assay, DAPI staining, flow cytometry analysis, RT-PCR, and Western blot analysis. RESULTS: At a concentration of 1.0 mg/mL, DCME inhibited the growth of HCT-116 cells by 84%, which was higher than at concentrations of 0.5 and 0.25 mg/mL. Chromatin condensation and formation of apoptotic bodies were observed in cancer cells cultured with DCME as well. In addition, DCME induced significant apoptosis in cancer cells by upregulation of Bax, caspase 9, and caspase 3, and downregulation of Bcl-2. Expression of genes commonly associated with inflammation, NF-kappaB, iNOS, and COX-2, was significantly downregulated by DCME. DCME also exerted an anti-metastasis effect on cancer cells as demonstrated by decreased expression of MMP genes and increased expression of TIMPs, which was confirmed by the inhibition of induced tumor metastasis in colon 26-M3.1 cells in BALB/c mice. CONCLUSIONS: Our results demonstrated that D. candidum had a potent in vitro anti-cancer effect, induced apoptosis, exhibited anti-inflammatory activities, and exerted in vivo anti-metastatic effects.
Animals ; Apoptosis ; Asia ; Asian Continental Ancestry Group ; Blotting, Western ; Caspase 3 ; Caspase 9 ; Chromatin ; Colon ; Colonic Neoplasms ; Down-Regulation ; Flow Cytometry ; HCT116 Cells ; Humans ; Inflammation ; Methanol ; Mice* ; Neoplasm Metastasis ; NF-kappa B ; Up-Regulation

Objective:To investigate the effects of exposure to ambient particulate matter (PM) and polycyclic aromatic hydrocarbons (PAHs) on systemic oxidative stress biomarkers in chronic obstructive pulmonary disease (COPD) patients.Methods: A panel of forty-five diagnosed and stable COPD patients, whose residences were within 5 kilometers from Peking University Health Science Center (PKUHSC), were recruited and followed up twice between November 2014 and May 2015.The lung function index percentage of forced expiratory volume in 1 second (FEV1) to predicted value (FEV1%pred), was measured to reflect the severity of COPD patients.The systemic oxidative stress biomarkers malondialdehyde (MDA) and 8-hydroxy-2′-deoxyguanosine (8-OHdG) in their urine samples were measured using high performance liquid chromatography (HPLC) and enzyme-linked immunosorbent assay (ELISA), respectively.Concentrations of ambient air pollutants and levels of meteorological factors were measured continuously through the air pollution-monitoring station located in PKUHSC.PM2.5 samples, which were used for measuring the concentrations of PAHs, were collected by PM2.5 high volume air sampler.We constructed linear mixed-effects models, including single-pollutant model, 2-pollutant models and stratification analysis, to estimate the effects of air pollutants on urinary MDA and 8-OHdG after adjusting for confounding factors.Results: In our COPD-patient panel, the associations between ultrafine particles (UFP) and PAHs and urinary MDA were statistically significant at lag2 (P<0.05).For an interquartile range (IQR) increase in UFP and PAHs, respective increases of 28% (95%CI: 4%-57%) and 36% (95%CI: 4%-77%) in urinary MDA were observed, and the effects became stronger after adjusting for the concentration of black carbon (BC).The COPD patients were divided into 2 groups stratified by FEV1%pred.Most air pollutants had stronger effects of systemic oxidative stress in the COPD patients of FEV1%pred≥50%.In this group, we observed that an IQR increase in UFP was associated with a 98% (95%CI: 38%-186%) increase in urinary MDA, and an IQR increase in BC, UFP and PAHs were associated with respective increases of 87% (95%CI: 32%-166%), 69% (95%CI: 24%-130%) and 156% (95%CI: 66%-294%) in urinary 8-OHdG.We didn't find significant associations between fine particulate matter (PM2.5) and urinary oxidative stress biomarkers.Conclusion: Our results suggested that exposure to air pollutants, especially UFP and PAHs, was responsible for exacerbation of systemic oxidative stress in COPD patients.Most air pollutants had stronger effects of systemic oxidative stress in mild to moderate COPD patients.

Objective To explore the value of DWI and 1 H-MRS in evaluation of malignant degree of primary osteosarcoma.Methods A retrospective analysis was performed in 41 patients with osteosarcoma confirmed pathologically and examined with DWI and 1 H-MRS.The b values were 0,400,800,1 000 and 1 500 s/ mm2 in DWI,respectively.ADC map was obtained,and ROIs were selected to measure ADC values.Eighteen cases which had stable spectral lines in MRS were selected and analyzed.The spectral lines,Choline (Cho),Creatine (Cr) and Cho/Cr ratio were obtained.Analysis of ADC values of high and low grade osteosarcoma were performed with Kruskal-Wallis H test,and Cho/Cr values were analyzed with Mann-Whitney U test.Results When b values were 400,800,1 000 s/mm2,ADC values of high and low grade osteosarcoma were significantly different (x2=6.26,4.37,4.87,P=0.01,0.04,0.03).When b value was 1 500 s/mm2,ADC value of high and low grade osteosarcoma had no statistical difference (x2 =2.39,P=0.12).Moreover,when 400 s/mm2 was taken as the optimal b value,the specificity and sensitivity of ADC value for identifying high grade osteosarcoma was 73.30 ％ and 72.70 ％,respectively.Cho/Cr values between high and low grade osteosarcoma were significantly different (Z=-2.45,P＜0.05).As a diagnostic parameter,the best threshold value of Cho/Cr was 2.21.And the specificity and sensitivity for diagnosing high grade osteosarcoma was 75.00 ％ and 92.90 ％,respectively.Conclusion ADC (b=400 s/mm2) and Cho/Cr values may be helpful to identifying malignant degree in osteosarcoma.

Returning to work is an important evaluation index for the recovery of physical function in patients with myocardial infarction. With the incidence of myocardial infarction in younger population and the prolongation of working age, young and middle-aged patients with myocardial infarction may become a population that needs to be paid attention to returning to work. This article reviews the status quo, influencing factors, evaluation tools and related intervention studies of young and middle-aged patients with myocardial infarction returning to work at home and abroad, aiming to provide a reference for the follow-up research of young and middle-aged patients with myocardial infarction returning to work.

Objective To investigate the mediating role of physical activity and negative psychological state be-tween sleep quality and cognitive function in stroke patients.Methods A total of 119 patients who were admitted and di-agnosed with acute ischemic stroke(AIS)in Zibo Central Hospital from January 2021 to December 2022 were selected,and a self-made scale was used to conduct a questionnaire survey.A Pearson correlation analysis was used to investigate the correlation of physical activity and negative emotions with sleep quality and cognitive function,and the hierarchical re-gression method was used to investigate the mediating role of physical activity and negative emotions.Results The 117 patients had a Pittsburgh Sleep Quality Index of(8.39±1.20).There were significant differences between the two groups in sex,age,physical activity score,Hospital Anxiety and Depression Scale score,and Montreal Cognitive Assessment Scale score(P<0.05).The Pearson correlation analysis showed that sleep quality score was negatively correlated with physical activity score and cognitive function score(r=-0.677 and-0.750,P<0.001)and was positively correlated with negative emotion score(r=0.585,P<0.001).The hierarchical regression analysis showed that physical activity and nega-tive emotions had a mediating role between sleep quality and cognitive function(P<0.05).Conclusion AIS patients have poor sleep quality after 30 days.There is a good correlation between sleep quality and physical activity/negative emo-tions/cognitive function,and physical activity and negative emotions plays a mediating role between sleep quality and cogni-tive function.