AIM: To study the feasibility, security a nd validity of the intrapericardial injection with a trans-diaphragmatic approach in chronic heart failure rats and its value in the study of gene therapy for hea rt diseases, and further investigate whether adeno-associated virual gene transf er of sarcoplasmic reticulum Ca 2+-ATPase (SERCA2a) gene can improve ven tricular function in chronic heart failure (HF) rats. METHODS: An animal model of heart failure was obtained by creati ng descending aortic constriction in rats. Recombinant adeno-associated virus, c arrying enhanced green fluorescent protein (EGFP) gene and recombinant adeno-ass ociated virus carrying SERCA2a gene, were respectively injected into pericardium of heart failure rats in different groups (group HF+EGFP and group HF+SERCA2a) by intrapericardial injection with a trans-diaphragmatic approach. After 30 days , hemodynamic parameters were measured and analyzed. Cryosection was analyzed by fluorescence microscopy to examine the expression of green fluorescent protein, and Western blotting was performed to detect the expression of SERCA2a. RESULTS: Green fluorescence was detected in cryosection of the h earts in all rats in group HF+EGFP and the expression of green fluorescent prote in was ubiquitously. The expression of SERCA2a in all rats in group HF+SERCA2a w as more than those in group HF and group HF+EGFP. And overexpression of SERCA2a improved the systolic and diastolic function of heart failure rats significantly and the hemodynamic parameters were similar with those of the controls. CONCLUSION: The intrapericardial injection with a trans-diaphrag matic approach suggests a simple, safe, efficient and cheap technique for the ge ne therapy of chronic heart failure. Gene thransfer of SERCA2a may be a new appr oach for the treatment of chronic heart failure.

Objective To investigate the prevalence of overweight and obesity and the nutritional status of dietary calcium intake in school-age children in Shanghai and to analyze the relationship between dietary calcium and bone mineral density (BMD).Methods A cross-sectional study was conducted in the second and third grade students of 10 primary schools in Yangpu District and Baoshan District in Shanghai. Food frequency questionnaire was used tu estimate the relative intake of dietary calcium of students in 6 primary schools in Baoshnn District. Quantitative ultrasound bone density examination was used to estimate the calcaneal BMD.The relation-ship between calcium intake and BMD was analyzed.Results The average calcium intake was 647 mg/d among ested students.BMD was not significantly different among different age groups (F=1.595,P=0.173),while Z-score was significantly different among different age groups (F=16.02,P=0.000).The total calcium intake was not correlated with BMD or Z-score.The percentage of dairy calcium intake was positively correlated with BMD (r=0.097,P=0.015)and Z-score (r=O.117,P=0.003),the percentage of non-dairy calcium intake and the non-dairy calcium intake were negatively correlated with BMD (r=-0.097,P=0.015) and Z-score(r=-0.110,P=0.006).When the total dietary calcium intake was ≥600 mg/d,BMD and Z-score significantly higherin students with high dairy calcium intake than those with low dairy calcium intake (P<0.05).When the total dietary calcium intake was <600mg/d,BMD and Z-score were higher in students with high dairy calcium in-take than in those with low dairy calcium intake,although no statistical significance was noted (P>0.05).Conclusions BMD and Z-score increase along with the increase of dairy calcium intake.Such effect becomes even more obvious when the total dietary calcium intake is ≥600 mg/d.

Objective To explore the role of defoamer enema combined with bellym assage on the colonic preparation in elderly patients with constipation. Methods One hundred patients were divided into two groups by random number table method, the experimental group and the control group with 50 cases each. Patients in the control group were told to drink polyethyleneglycolelectrolytesolution 3000 ml. Patients in the experimental group were told to drink polyethyleneglycolelectrolytesolution 2000 ml. After medicinepre paration, the patients of experimental group were given defoamer enema. After that, they were undertaken counterclockwise massage for10 mins, then massageing clockwise until defecation. Results 14 patients with oral catharsis drugs failed to give up check, 46 cases of intervention group and 40 cases of control group finally complete intestinal preparation and colonoscopy. Intervention group patients after bowel preparation before the incidence of abdominal distension, abdominal pain were 6.52% (3/46), 8.70%(4/46), lower than the control group 65.00% (26/40), 25.00% (10/40), the difference was statistically significant (χ2= 32.74, 4.17, P< 0.05). Percent of pass was 65.22%(30/46) for intestinal preparation intervention group, significantly higher than the control group 35.00% (14/40), the difference was statistically significant (χ2= 7.82, P< 0.05). Intervention group intestinal cleanliness ratings of Ottawa total score was 4.00 (4.00), which was lower than the control group 7.00 (4.50), the difference was statistically significant (Z= 3.80, P< 0.05). Endoscopic check process, the intervention group arrived at the terminal ileum and mirror back time were 7.00 (3.00) and 9.00 (1.00) min, were less than 9.00(6.50) and 10.50 (3.00) min in the control group, the difference was statistically significant (Z= 2.09, 4.53, P< 0.05). Intervention group of colons polyps detection rate was 67.39% (31/46), higher than that of control group 30.00% (12/40), the difference was statistically significant (χ2 = 11.97, P< 0.05). Conclusions The bowel preparation with defoamer enema will enhance the intestinal tract cleaness and the detection rate of polyps in elderly patients with constipation.

Objective To evaluate the effect of dexmedetomidine on the expression of antisense hypoxia-inducible factor-1α (aHIF-1α) during brain injury after asphyxial cardiac arrest and resuscitation in rats.Methods Twenty-four pathogen-free healthy male Sprague-Dawley rats,weighing 250-280 g,were divided into 3 groups (n =8 each) using a random number table:sham operation group (group S),cardiac arrest and resuscitation group (group R) and dexmedetomidine group (group D).Asphyxial cardiac arrest and resuscitation were performed in R and D groups.The rats were tracheally intubated without clipping the trachea in group S.Dexmedetomidine 4 μg/kg was injected via the tail vein at 5 min before clipping the trachea in group D,while the equal volume of normal saline was given instead in S and R groups.Neurological deficit was assessed and scored (NDS) at 12,24,48 and 72 h after recovery of spontaneous circulation (T1 4).The rats were sacrificed after assessing neurological deficit at T4,brains were removed and hippocampi were isolated for determination of cell apoptosis (by TUNEL),HIF-1α expression (by Western blot) and expression of HIF-1α and aHIF-1α mRNA in hippocampal tissues (using polymerase chain reaction).Apoptosis rate was calculated.Results Compared with group S,the NDS at each time point and apoptosis rate of hippocampal neurons at T4 were significantly increased,and the expression of HIF-1α protein and mRNA and 5'aHIF-1α mRNA was up-regulated in R and D groups (P＜O.05).Compared with group R,the NDS at T2.4 and apoptosis rate of hippocampal neurons at T4 were significantly decreased,the expression of 5'aHIF-1α mRNA was down-regulated,and the expression of HIF-1α protein and mRNA was up-regulated in group D (P ＜ 0.05).Conclusion The mechanism by which dexmedetomidine reduces brain injury after asphyxial cardiac arrest and resuscitation is related to down-regulation of 5'aHIF-1α expression in rats.

Objective To study the impact of establishment of lipopolysaccharide ( LPS )-induced rat model of chronic obstructive pulmonary disease (COPD)on the function of multidrug resistance-associated protein 1(MRP1)in the rat bronchial epithelium .Methods Using intratracheal instillation of LPS to establish COPD rat model .8-week old healthy male Wistar rats were divided into 3 groups (10 rats in each group ):(1) Normal control;(2) Modeling for 14 days after LPS instillation;(3) Modeling for 28 days after LPS instillation.Pulmonary function and the concentration of phenol red in bronchoalveolar lavage fluid ( BALF) and plasma were measured .The ratio of phenol red concentration in BALF/plasma was used as an index of the MRP 1 function in the rat bronchial epithelium and the expression of MRP 1 in the bronchial epi-thelium was also observed by immunohistochemistry .Results Compared with the normal group , the pulmonary functions of the rats in the model groups were significantly reduced along with the modeling progress .After intravenous administration of phenol red, the ratio of phenol red concentration in BALF/plasma was gradually reduced , and the expression of MRP1 in the bronchial epithelium was significantly decreased .Conclusions COPD rat model can be established by intratracheal LPS instillation, and the function of MRP1 in bronchial epithelium was gradually reduced along with the modeling progress .

AIM:To investigate whether mitochondrial membrane potential (ΔΨm ) and the mitochondrial ap-optotic pathway are involved in the protective mechanism of Panax quinquefolium saponin ( PQS) against cardiomyocyte ap-optosis after ischemia/reperfusion ( I/R) injury in rat myocardium .METHODS: Ninety healthy male SD rats were ran-domly divided into sham group , I/R group, PQS (200 mg· kg -1 · d-1 ) +I/R group, cyclosporine A ( CsA) group, CsA (10 mg· kg-1 ) +I/R group and PQS +CsA +I/R group.The model of myocardial I/R injury in vivo was established by ligating the left anterior descending artery ( LAD) for 30 min followed by 120 min of reperfusion in the rats .The serum ac-tivity of lactate dehydrogenase ( LDH ) was measured by automatic chemistry analyzer .The myocardial infarct size was measured by Evans blue and 2,3,5-triphenyltetrazolium chloride (TTC) staining.Cardiomyocyte apoptosis was detected by in situ TDT-mediated dUTP nick end labeling (TUNEL).The protein levels of Bcl-2, Bax, cleaved caspase-3 and cytosol-ic cytochrome C were determined by Western blotting .ΔΨm was measured by laser scanning confocal microscopy and fluo-rescence microplate reader .RESULTS:Compared with I/R group, the serum content of LDH ,the infarction size in PQS+I/R group, CsA+I/R group and PQS +CsA+I/R group and the myocardial apoptotic index were decreased .Compared with I/R group, the fluorescence intensity of mitochondria after JC-1 staining was enhanced in PQS +I/R group, CsA+I/R group and PQS+CsA+I/R group, and the relative fluorescence units (RFU) ofΔΨm were improved in those 3 groups. In PQS+I/R group, CsA+I/R group and PQS+CsA+I/R group, the protein expression of Bcl-2 was increased, and that of Bax was decreased compared with I/R group.Moreover, in those 3 groups, the protein levels of cleaved-caspase-3 and cytosolic cytochrome C were decreased compared to I /R group, respectively.CONCLUSION:PQS attenuates myocardial injury and cardiomyocyte apoptosis during I /R, and the protective mechanisms of PQS were associated with the modulation ofΔΨm and the inhibition of mitochondrial apoptosis pathway .

AIM:To study the effect of Panax quinquefolium saponin (PQS) on cardiomyocyte apoptosis in-duced by thapsigargin ( TG) .METHODS:Primary cultured cardiomyocytes from neonatal SD rats were divided into con-trol group, TG group, PQS (40 mg/L, 80 mg/L and 160 mg/L) +TG group, si-PERK+TG group, and mock+TG group.The cells were treated with 1 μmol/L TG for 24 h to induce apoptosis.The PERK gene in the cardiomyocytes was knocked down by RNAi.The cell viability was detected by CCK-8 assay.Apoptosis was analyzed by flow cytometry.Wes-tern blotting was used to determine the expression of ERS molecules GRP78, CRT, ATF4 and CHOP, anti-apoptosis pro-tein Bcl-2 and pro-apoptosis protein Bax.RESULTS: Compared with control group, TG significantly and the apoptosis, reduced the cell viability (P<0.05), increased the phosphorylation of PERK and eIF2α, increased the expression of GRP78, CRT, ATF4, CHOP and pro-apoptosis protein Bax, and decreased the expression of anti-apoptosis protein Bcl-2 (P<0.05).Compared with TG group, PQS treatment (160 mg/L) significantly reduced the apoptosis and increased the cell viability (P<0.05).All the 3 different concentrations of PQS significantly increased the expression of anti-apoptosis protein Bcl-2 and reduced the expression of pro-apoptosis protein Bax (P<0.05) in a dose-dependent manner.PQS pre-treatment and knockdown of PERK both reduced the protein levels of GRP78, CRT, PERK, p-PERK, eIF2α, p-eIF2α, ATF4, CHOP and pro-apoptosis protein Bax, and increased the expression of anti-apoptosis protein Bcl-2 (P<0.05). CONCLUSION:PQS at concentration of 160 mg/L attenuated cardiomyocyte apoptosis induced by TG.PQS had the simi-lar effect as PERK knockdown on cardiomyocyte apoptosis.The mechanism may be associated with inhibiting PERK-eIF2α-ATF4-CHOP pathway of ERS-related apoptosis.

Objective To investigate the risks of self-rated health in the ≥55-year elderly in Beijing and the occurrence of stroke.Methods The subjects (n=2 101;aged ≥55) from Beijing longitudinal study of aging (BLSA) were collected by Xuanwu Hospital,Capital Medical University from January 1992 to December 2016.One hundred and twenty-one subjects with stroke at baseline and 92 with incomplete information were excluded,and finally,1 888 elderly patients without cerebrovascular disease at baseline were included in the analysis.Based on the actual situation,the self-rated health was to identify an item that matched their current state from good,general to poor.The deadline for the survey was December 31,2012.The competitive risk model was used to assess the health self-rated status and the risk of stroke.Non-stroke deaths,including cancer and car accidents were treated as competitive events.Results Of the 1 888 subjects enrolled,946 (50.1%) self-rated health were good,616 (32.6%) were general,and 326 (17.3%) were poor;438 (23.2%) had stroke,751 (37.8%) had non-stroke death,and 699 (37.0%) were right censored data.Using the competing risk model and adjusting the age,sex,living area,marital status,education level,smoking,alcohol consumption,physical exercise,hypertension,diabetes mellitus,coronary heart disease,and body mass index,the occurrence of stroke in patients with poor self-rated health was 1.44 times (95%CI 1.11-1.87,P<0.01) as good as those who were good.Conclusion In the self-rated health of the elderly ≥55 years old in Beijing,the people with poor self-rated health increased the occurrence of stroke after considering the competitive risks.

Objective To investigate the effects of physical exercises on cardiac function and plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) levels in hypertensive patients combined with diastolic cardiac dysfunction.MethodsA total of 66 essential hypertension patients who had abnormal left ventricular relaxation and normal systolic function were assigned to the intervention group ( n =33 ; doing physical exercises once a day,5 days a week) or control group (n =33 ).All the patients received standard treatment.At 6 months,body weight,blood pressure,heart rate,NT-proBNP,and echocardiography were measured.ResultsAt 6 months,body weight [ (68 ± 7 ) kg vs (72 ± 8 ) kg ],systolic blood pressure [ (135.4 ±5.1) mm Hg (1 mm Hg =0.133 kPa) vs (141.9 ±5.2) mm Hg ],diastolic blood pressure [ (81.1 ±4.0) mm Hg vs (84.7 ±4.6) mm Hg],New York Heart Association class (1.4 ±0.3 vs 1.8 ±0.4),NT-proBNP level [ (526 ± 126 ) ng/L vs (741 ± 189 ) ng/L] were significantly decreased in the intervention group when compared with the control group ( all P ＜ 0.05 ) although left ventricular ejection fraction (LVEF) (62.9 ±6.7 vs 59.0 ±5.6) and E/A ratio ( 1.1 ±0.3 vs 0.9 ±0.3) were significantly increased ( both P ＜ 0.05).ConclusionPhysical exercises could play a role in reduced blood pressure and body weight and improved cardiac function in hypertensive patients with diastolic cardiac dysfunction.

OBJECTIVEThe interrelation of yield and agronomic traits of Gentiana rigescens was studied for the germplasm and breeding variety of this species.

METHODTwelve agronomic traits, root diameter, root length, root number, root biomass, stem diameter, plant height, the first branch number, leaf length, leaf width, leaf length/leaf width ratio, calyx length, and calyx number of G. rigescens from 26 wild populations in Yunnan were determined for correlation analysis, multiple stepwise regression analysis and path analysis.

RESULTCorrelation analysis showed that there were significantly positive correlation between the traits of aboveground part and the length, diameter, number, and biomass of the root. Multiple stepwise regression analysis showed that length, width, and number of root, plant height, the first branch number, and the calyx number were the main factors that affected the root biomass. Path analysis showed that the diameter, length, and number of the root, the stem height, and the first branch number had a direct positive effect on the root biomass.

CONCLUSIONThe traits, such as high and strong stem, high number of first branch number and shrubby shape could be selected for the breeding and high yielding of G. rigescens.