Hepatitis C virus(HCV)is the major etiology of non-A,non-B hepatitis.At present,neither a vaccine nor any effective therapy is available.Multi-epitope DNA vaccine(minigenes/epigenes)is a novel nucleic acid vaccine which can induces high effective cellular and humoral immune responses and has good perspective in clearing Hepatitis C virus through screening and assembling optimal antigen epitope genes(T cell、B cell epitope included).It is advanced in covering more HCV subtypes,inducing comprehensive anti-HCV immune responses and reducing the negative influence caused by irrelevant,disturbing and suppressive sequence as far as possible through selecting the most potential protective epitopes in DNA vaccine design.The recent research progress on HCV compound multi-epitope DNA vaccine and future prospects were reviewed.

Objective To evaluate the value of reducing radiation dose with decreased size of ECG-pulsing windows and influence on image quality in dual-source CT coronary angiography. Methods 120 patients with stable heart rate(HR) were divided into four groups according to HR and the rang of ECG-pulsing windows in dual-source CT coronary angiography: HR ＜ 70 bpm and 61% ～ 77% R-R interval of ECG-pulsing windows, HR ＜ 70 bpm and 25% ～ 80% R-R interval of ECG-pulsing windows,HR ＞ 80 bpm and 31% ～47% R-R interval of ECG-pulsing windows, and HR ＞ 80 bpm and 25% ～ 80%R-R interval of ECG-pulsing windows was employed, respectively. The radiation dose parameters were recorded and image quality scores were performed. The image quality and radiation dose between two slow HR groups and between two fast HR groups were compared respectively. Result The effective doses were (7.06 ± 2. 13 ), ( 11.34 ± 3.65 ), ( 6. 67 ± 1.97 ) and ( 9. 92 ± 3. 15 ) mSy for four groups, respectively.The effective dose was decreased by 37.74% for slow HR and by 32. 76% for fast HR using narrow ECG-pulsing windows. There was no difference on image quality between two slow HR groups and two fast HR grouvs. Concluslons The proper application of narrow ECG-pulsing windows can reduce radiation exposure significantly to stable slow or fast HR patients in dual-source CT coronary angiography withont sacrificing the image quality.

Objective:A novel tuberculosis DNA vaccine integrating siRNA,fusional antigen and hIL-12 triple expression units was constructed in our laboratory.Methods:To evaluate the independent expression of the three expression units,two eukaryotic expression plasmid pVAX1-siRNA-PVAE[EGFP]-hIL12 with TB fusional gene Ag85B-ESAT6(PVAE) and enhanced green fluorescent protein(EGFP),and pVAX1-siEGFP-PVAE[EGFP]-hIL12 with siRNA coding sequence targeting EGFP instead of Mcl-1L were constructed.Then two plasmids were used to transfer human embryonic kidney 293 cells.Based on EGFP report gene,it was demonstrated that siRNA expression unit and fusional antigen gene were independently expressed.Results:The hIL12 expression at 48h and 72h post transfection were also detected by ELISA analysis up to 1571.63pg/ml and 2392.25pg/ml respectively in the cell culture fluid.Conclusion:The results demonstrated that the novel DNA vaccine with siRNA,TB fusional antigen and hIL12 three expression units in the same plasmid frame is successfully constructed and independently expressed in eukaryotic cells.It laid a foundation for further animal model study on anti-tuberculosis effects of this novel DNA vaccine.

The inverted terminal repeat (ITR) is the only cis element of AAV genome essential for rAAV rescue, replication and packaging. It is prone to mutation or loss when it is latent in host cell or in plasmid. Plasmids with different ITR types were cloned to compare the influence of ITR types on the AAV packaging and infectivity. The vector plasmids were transformed the competent SURE cells to get different colonies. The ITR types of plasmids were screened by digestion with SmaⅠ. AAV vector plasmid pScGFPud has two ITRs at both ends of AAV genome and plasmid pScGFPu has only one ITR at upstream end of AAV genome. When the two plasmids were co-transfected 293 cells to prepare rAAVs, 1.08?1013 viral particles (AAV1-GFPud) were produced from 20-dishes of 293 cells cotransfected with plasmid pScGFPud, 4.28?1012 viral particles (AAV1-GFPu) were produced from 20-dishes of 293 cells cotransfected with plasmid pScGFPu. Virus AAV1-GFPud infected 293, HeLa and NCI H446 cells more efficiently than did virus AAV1-GFPu. This suggests that defect ITRs in AAV genome is deleterious to AAV packaging and AAV infectivity and vector with complete ITRs is favorable to the yield and activity of rAAV.

Objective:To investigate the efficacy and safety of dexmedetomidine and dexamethasone in inhibiting opioid-induced cough (OIC) during general anesthesia induction in patients with gynecological tumors.Methods:A total of 180 patients who were scheduled for elective gynecological tumor surgery under general anesthesia in Shanxi Provincial Cancer Hospital from March to November 2019 were selected. They were randomly divided into blank control group, dexmedetomidine group and dexamethasone group according to the random number table method, each group had 60 cases. Firstly, all patients had a 10-minute rest (T 0) after they entered the operate room. Treatment before general anesthesia induction:dexmedetomidine group was pumped dexmedetomidine 0.5 μg/kg (diluted to 10 ml with 0.9% NaCl injection) using an electronic infusion pump; dexamethasone group was injected intravenously dexamethasone 10 mg; blank control group was pumped with 10 ml 0.9% NaCl injection. The pumping was finished within 5 minutes, and the end time of pumping was denoted as T 1. Induction of general anesthesia was performed 5 minutes after the end of pumping: firstly, sufentanil was given intravenously at 0.3 μg/kg, and the injection was finished within 5 seconds (T 2). Two minutes after sufentanil injection (T 3), cis-atracurium 0.3 mg/kg and propofol medium/long-chain injection 2 mg/kg were sequentially injected. Then preoxygenation, endotracheal intubation and mechanical ventilation were implemented in turn. One minute after intubation was recorded as T 4. The incidence and severity of cough in patients within T 2-T 3 of each group were recorded, as well as the incidence of tachycardia, bradycardia, hypertension, hypotension, respiratory depression and myotonia during T 1-T 4. Results:The incidence of OIC in the dexmedetomidine group (10.0%, 6/60) and dexamethasone group (8.3%, 5/60) was lower than that in the blank control group (33.3%, 20/60), and the difference among the three groups was statistically significant ( χ2 = 16.445, P < 0.01), while there was no significant difference in the incidence of OIC between the dexmedetomidine group and the dexamethasone group ( P > 0.05). The incidence of sinus bradycardia in the dexmedetomidine group (16.3%, 10/60) was higher than that in the blank control group (0, 0/60) and dexamethasone group (8.4%, 1/60), and the difference was statistically significant ( P < 0.05). Respiratory depression and myotonia did not occur in the three groups. Conclusions:Pretreatment with dexmedetomidine or intravenous dexamethasone before anesthesia induction can effectively reduce the incidence of OIC in patients with gynecological tumors, and there is no significant difference between the effects of the two drugs. The incidence of sinus bradycardia increases significantly after dexmedetomidine infusion.

Objective To evaluate blood flow structure and quantify the variation of the flow within left ventricle,assess the impact of chronic heart failure(CHF) by vector flow mapping(VFM).Methods Twenty-seven patients with chronic heart failure and thirty controls were involved.The flow vector images on the section plane of the flow within the left ventricle were acquired by VFM.Time-flow(T-F) curve and all other peak systolic and diastolic flow curve include normal velocity profile,parallel velocity profile,vector profile,flow profile were analyzed by DSA-RS1 program.Results Ventricular ejection peak S,rapid ventricular filling peak E and atrial systole peak A were relatively lower at basal and middle segments in CHF group than normal control group.Normal velocity profile,velocity profile,flow profile at peak S and E were lower at basal and middle segments in CHF group than normal control group.Conclusions VFM technology could provide quantitative and intuitive information to demonstrate the flow structure of the ventricle and evaluate the cardiac function in patients with CHF.

Objective To investigate the prevalence status and risk factors of mild cognitive impairment (MCI) in old people in pension facilities in Chongqing, China. Methods From October to November, 2015, the people above 60 years old in four pension facilities in Chongqing were cluster sampled. They were interviewed with Mini-Mental State Examination, Instrumental Activity of Daily Living Scale, self-made general situation questionnaire for the elderly and the Geriatric Depression Scale. Results There were 25 MCI patients identified in 84 old people selected, with the prevalence of 29.76%. MCI prevalence was different with the factors such as genders, educational levels, economical status, smoking, eating freshwater fish frequently, participating in outdoor activities frequently, playing Mahjong/chess/cards fre-quently, watching TV/listening to broadcast/reading newspaper frequently, participating in social activities frequently, having religious be-liefs, suffering high blood pressure (χ2>4.271, P<0.05). Based on multivariate Logistic regression analysis, male was the protective factor (OR=0.055, 95%CI:0.005~0.618), while high blood pressure was the risk factor (OR=8.281, 95%CI:1.278~53.646) to the MCI. Conclu-sion MCI is prevalent in old people in pension facilities in Chongqing, which needs intervention targeted to the risk foctors.

Objective To investigate the pathological manifestations of acute?stage radioactive myocardial damage and related mechanisms using an experimental animal model. Methods A total of 12 adult male Sprague?Dawley rats were randomly divided into control group and radiation group. In the radiation group, the model of radiation?induced myocardial damage was established by the irradiation of the anterior myocardial territory with 6?MV X?ray at a single dose of 20 Gy, and at 14 days after irradiation, HE staining was used to observe the morphological changes of cardiomyocytes and intercellular matrix, and Masson staining was used to observe the distribution of collagen fibers. The collagen volume fraction ( CVF) was used for semi?quantitative analysis. ELISA was used to measure the activity of total superoxide dismutase ( T?SOD) and the concentration of malondialdehyde ( MDA ) , and Western blot was used to measure the expression of the fibrosis marker protein collagen type I ( COL?1 ) and the endoplasmic reticulum stress?related proteins GRP78 and CHOP. The t?test, t ’?test, or nonparametric rank sum test was used for comparison between groups. Results At 14 days after local irradiation of the heart, the radiation group had disordered arrangement of cardiomyocytes, marked edema, rupture of some cardiomyocytes, mild karyopyknosis in cardiomyocytes, and infiltration of inflammatory cells in the myocardial interstitium, compared with the control group. Collagen fibers in the myocardial tissue were mainly distributed in the perivascular area and cardiomyocyte interstitium. The radiation group had a significantly higher CVF than the control group ( 11?35% vs. 5?23%, P=0?000 ) and a significant increase in the expression of COL?1 compared with the control group ( P=0?000) . The radiation group had significant increases in the activity of T?SOD and the concentration of MDA in the myocardial tissue compared with the control group ( T?SOD:156?61 U/mgprot vs. 137?06 U/mgprot, P= 0?042;MDA:2?36 nmol/mgprot vs. 1?31 U/mgprot, P=0?007) . Compared with the control group, the radiation group showed significant increases in the expression of endoplasmic reticulum stress?related proteins GRP 7 8 and CHOP ( P= 0?037 and 0?009 ) . Conclusions The pathological manifestations of acute?stage myocardial damage include degeneration of cardiomyocytes, inflammatory exudation in the interstitium, and deposition of collagen in the perivascular area and myocardial interstitium. Myocardial fibrosis can be observed in the acute stage of radiation?induced myocardial damage, and the mechanism may be related to oxidative stress and endoplasmic reticulum stress induced by radioactive rays.

Objective:To construct a DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the anti-HBV siRNA genes, and to express this DNA vaccine in HepG2 cells. Methods:The HBV multi-epitope antigen gene was designed and synthesized before it was fused with enhanced green fluorescent protein(EGFP) gene, and cloned into the multi-clone site(MCS) of the eukaryotic expression vector pVAX1. The expressinig units of hIL-12 and siRNA were cloned into the BspH I and Mlu I site of pVAX1 respectively. Then the recombinant plasmid pVAX1-siHBV-HB-EGFP-hIL12 was transiently transfected HepG2 cells. The expression of HBV compound multi-epitope gene was observed through EGFP report gene. The expression of hIL-12 was analyzed by ELISA and the effects of anti-HBV siRNA was confirmed with rtPCR . Results: The analysis of enzyme digestion and sequencing both demonstrated that the trible-expressing HBV DNA vaccine has been constructed successfully. The green fluorescent image was detected in the transfected cells which could confirm the expression of the multi-epitope antigen gene. The amount of hIL-12 secretion was 1289pg/ml in supernatant at 48h after transfection and 1712pg/ml at 72h after transfection. The mRNA amount of HBV S gene, which was the siRNA target, had been obviously knockdown. Conclusion: The DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the siRNA genes was constructed and transiently expressed in HepG2 cells, and siRNA had shown us a good anti-HBV effect. It laid a foundation of further study on anti-HBV effect of the new DNA vaccine.

Objective To investigate the prevalence and risk factors of mild cognitive impairment (MCI) among the old people in Chongqing, China. Methods From October to November, 2015, 1850 persons more than 60-year-old in Chongqing were cluster sampled. They were investigated with self-made general situation questionnaire, Mini-Mental State Examination (MMSE), Instrumental Activity of Daily Living Scale (IADL) and Geriatric Depression Scale (GDS) through face-to-face interview. Enumeration data were tested withχ2 test and multiple Logistic regression. Results MCI prevalence was 11.73%in the old people in Chongqing, and various with the residential re-gions, ages, marital status, literacy, career, smoking, drinking, seafood-eating, egg-eating, exercising, intensive labor, low intensity of labor, housework, outdoor activities, Mahjong/chess/cards playing, TV-watching/radio-listening/newspaper-reading, social activities, obesity, hy-pertension, diabetes, stroke, hyperglycemia, cerebrovascular insufficiency and depression (χ2>4.092, P<0.05). According to multiple Logis-tic regression, living in rural areas, aging, celibacy, obesity, hypertension and depression were the risk factors of MCI;while middle school and above years of schooling, mental workers, smoking cessation, frequent seafood-eating and egg-eating, exercising, daily intensive labor and low intensity of labor, daily and sometimes housework, daily and weekly outdoor activities at least once, daily Mahjong/chess/cards playing and daily TV-watching/radio-listening/newspaper-reading were the preventing factors. Conclusion The morbidity of MCI is high in the old people in Chongqing. It is necessary to take effective intervention in view of the related factors of MCI as early as possible, to delay or prevent the development of MCI to dementia.