0 05), being ( 187 0?10 7) nmol/L in average. Moreo ver, it was not significantly different from the cells cultured at day 0, ei ther . However, [Ca 2+ ]i increased significantly(P

OBJECTIVE:To evaluate the rationality and validity of chemical medicine tablet production equipment cleaning pro-cedure. METHODS:Among several chemical medicines prepared by similar production technology as Metoprolol succinate sus-tained-release tablets,Captopril tablets,Isosorbide mononitrate tablet and Metformin hydrochloride tablet,Metoprolol succinate sus-tained-release tablets had strongest toxicity and were included in validation test. The production equipment was cleaned and disinfect-ed according to cleaning procedure. The point which was most difficult to clean could be wiped and sampled by using the cotton swab method. The detection limit and the limit of quantitation of the residue limits were verified as well as the recovery rate of wip-ing,in order to evaluate whether the results meet the requirements. RESULTS:The cotton swab method is adopted to wipe sample and detect the point which is most difficult to clean. The visible foreign body has not been found in each sampling point. The amount of residual drug is <29.75 μg/cotton bud,and microbial limits are <50 CFU/cotton bud,indicating test items are in line with the standard. CONCLUSIONS:The cleaning method can effectively clean the production equipment,and can effectively pre-vent product contamination and cross contamination to ensure the quality,efficacy and safety of the next batch of products.

BACKGROUNDTo explore the feasibility of cloning of the hepatocyte receptor interacting with the Pre S1 protein of HBV by two hybrid system.

METHODSYeast expression plasmids encoding fusion proteins of full length or portions of Pre S1 of HBV and DNA binding domain of yeast protein GAL4 were constructed and used to transform yeast reporter strain SFY526. Reporter gene product ?galactosidase activity was assayed as a measure of transcription activation in yeast. Mammalian expression plasmid encoding fusion proteins of full length Pre S1 and DNA binding domain of GAL4 was constructed and used to cotransfect hepatoma cell line Huh?7 together with CAT reporter plasmid. Cell extracts were assayed for CAT activity by thin?layer chromatography.

RESULTSThe fusion proteins of full length Pre S1 protein and GAL4 DNA binding domain present transcriptional activation function in yeast. The transcription activating sequence is localized to the 21 to 47 amino acids of Pre S1 protein Fusion proteins of full length Pre S 1 and GAL 4 DNA binding domain do not show transcriptional activation function in mammalian cells.

CONCLUSIONThe transcriptional activating sequence of HBVPre S1 protein in yeast overlaps the hepatocyte receptor binding site. The transcriptional activation function of HBV Pre S1 protein in yeast may prevent researchers?from using yeast two hybrid system to clone HBV receptor interacting with Pre S1 protein. However, the Pre S1 protein does not show transcriptional activation function in mammalian cells. Mammalian two?hybrid system may be a practical method to clone the HBV hepatocyte receptor interacting with Pre S1 protein.

DNA-Binding Proteins ; genetics ; Fungal Proteins ; genetics ; Hepatitis B Surface Antigens ; genetics ; Humans ; Protein Precursors ; genetics ; Recombinant Fusion Proteins ; genetics ; Transcriptional Activation ; Tumor Cells, Cultured ; Yeasts

Objective:To study the clinical and molecular characteristics of a family with familial hypercholesterolemia (FH) with LDLRAP1 and ABCG8 gene abnormality.Methods:In September 2020, one case of FH was included in Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine; peripheral venous blood samples of members of the family were collected to detect serum total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) indicators; use high-performance liquid chromatography to detect serum stigmasterol and sitosterol content; perform second-generation gene sequencing to detect gene mutations in probands and family members; use Pymol software to detect gene mutations point for pathogenicity analysis, and use Uniprot Modelling software to perform protein structure modeling.Results:The patient presented with anemia, multiple xanthomas and early-onset acute coronary syndrome. The coronary angiography showed severe coronary artery lesions; abdominal ultrasound showed splenomegaly; blood smear showed shaped erythrocytes and large platelets. The level of serum TC, LDL-C, stigmasterol and sitosterol was 8.54 mmol/L (2.3-5.7 mmol/L), 4.84 mmol/L (range of normal value 1.3-4.3 mmol/L), 44 μmol/L (1.0-10 μmol/L), 28 μmol/L (1.0-15 μmol/L), respectively; LDLRAP1 gene mutation was found: exon4 c.415C>T:p.Q139X; the truncated protein formed by this homozygous mutation lost multiple stable protein structure regions, which can not have a normal function. At the same time, ABCG8 gene mutations were also found: exon13 c.1895T>C (p.V632A) and exon8 c.1199C>A:p.T400K . Two cases of family members had a mild increase in HDL-C (Ⅱ5: 2.33 mmol/L, Ⅱ6∶2.96 mmol/L), 3 cases carrying the ABCG8 gene mutations had a slight increase in stigmasterol (Ⅱ8: 23 μmol/L, Ⅱ7: 24 μmol/L, Ⅰ2: 18 μmol/L) and sitosterol (Ⅱ8: 41 μmol/L, Ⅱ7: 33 μmol/L, Ⅰ2: 45 μmol/L), suggesting that its association with the concentration of plant sterols. Conclusions:FH patients with LDLRAP1 and ABCG8 gene abnormalities may have abnormal plant sterol concentrations, and their clinical manifestations are more complicated. Therefore, family history, LDL-C, plant sterol levels, and genetic test results should be considered comprehensively.

Objective To analyze the death attributable to smoking and impact of life expectancy in China in 2013.Methods According to the characteristics of different diseases,we calculated the population attributable fractions of different diseases,death and impact of life expectancy which caused by smoking,using direct method (current smoking rate as exposure levels)and indirect method (smoking impact ratio as exposure levels),based on data from both programs of death surveillance and Chinese chronic disease risk factor surveillance of 2013.Results In 2013,smoking caused around 1.59 million deaths which accounted for 17.38％ of all deaths in China.Constituent ratio of death caused by smoking in males (23.66％) was much higher than that in females (8.30％).However,in urban areas (17.24％),it was slightly lower than that in rural areas (17.51％).Constituent ratio of death caused by smoking in the eastern regions appeared the lowest (16.81％),with western regions the highest (17.91％).In 2013,lung cancer,COPD and ischemia heart disease were the top three diseases causing deaths that related to smoking,but the top three population attributable fractions were lung cancer,COPD and nasopharyngeal carcinoma in China.In 2013,smoking caused a reduction of 2.04 years of life expectancy loss in China,with males in the western regions the highest (3.05 years).Conclusion Smoking is still an important public health problem in China.Tobacco-control-targeted programs in the heavily involved areas could reduce the number of deaths from related diseases that caused by smoking.

Objective To estimate the deaths (mortality) and life expectancy that attributable to high blood pressure in people from different regions and gender,in China in 2013.Methods Data was from the ‘China Chronic Disease Risk Factor Surveillance 2013'and the ‘China National Mortality Surveillance 2013'.According to the comparative risk assessment theory,population attributable fraction (PAF) of high blood pressure by gender,urban-rural,east-central-west regions was calculated before the estimations on deaths (mortality) and life expectancy attributable to high blood pressure was made.Results In 2013,among the Chinese people aged 25 years old and above,the mean SBP was (129.48 ± 20.27) mmHg.High blood pressure [SBP＞(115 ± 6) mmHg] caused 20.879 million deaths and accounted for 22.78％ of the total deaths.SBP,deaths,mortality rate and standardized mortality rate that attributable to high blood pressure all appeared higher in men [(131.15 ± 18.73) mmHg,11.517 million,165.56/100 000 and 106.97/100 000,respectively] than in women [(127.79 ± 21.60) mmHg,9.362 million,141.99/100 000 and 68.93/100 000,respectively].SBP,deaths,mortality rate and PAF were all seen higher in rural [(130.25±20.66) mmHg,11.234 million,178.58/100 000 and 23.59％,respectively] than in urban [(128.58± 19.77) mmHg,9.645 million,132.87/100 000 and 21.54％,respectively] areas.However,levels of SBP were similar in the east,central or west regions,with attributable deaths,attributable mortality rate and PAF the highest as 7.658 million 179.93/100 000,and 26.72％ respectively.In 2013,among the Chinese people aged 25 years old and above,deaths caused by cardiovascular disease and chronic kidney disease attributable to high blood pressure were 19.912 million and 0.966 million,accounting for 52.31％ of the total deaths due to cardiovascular diseases and 62.11％ to the total chronic kidney diseases.The top three deaths attributable to high blood pressure were ischemic heart disease (6.656 million),hemorrhagic stroke (5.331 million) and ischemic stroke (3.593 million).When the effect of high blood pressure had been eliminated,the life expectancy per capita would have increased by 2.86 years old,with higher in women than in men (3.07 and 2.64 years old,respectively),higher in central than in east and west (3.48,2.56 and 2.58 years,respectively) areas,in rural than in urban (2.97 and 2.59 years,respectively) areas.Conclusions In 2013,the number of deaths attributable to high blood pressure was around 20.9 million,accounting for 22.78％ of the total deaths,and appeared higher in men than in women,in rural than in urban,in central than in east and west areas.The mortality burden induced by ischemic heart disease,hemorrhagic stroke and ischemic stroke was most serious since the high blood pressure brought about 2.86 years of lost in life expectancy.

Objective To quantitatively estimate the deaths and life expectancy losses attributable to diet high in sodium in China,and examine the gains and shifts under different control scenarios of sodium consumption.Methods Based on data from the cause-of-death through the National Mortality Surveillance System,and 24 hours urinary sodium values from Global Burden of Disease study on Chinese's estimates,population attributable fractions with the framework of comparative risk assessment were used to analyze the deaths and life expectancy losses due to diet high in sodium.The same methods were followed to examine the gains and shifts under different control scenarios of sodium consumption.Results In 2013,1 430 (940 for men and 490 for women)thousand deaths were attributable to diet high in sodium,accounting for 15.6％ (17.4％ for men and 13.0％ for women) of all-cause deaths in China,which causing 2.17 (2.49 for men and 1.71 for women) years of life expectancy loss.Diet with high sodium in 2013 caused 1 200,50 and 180 thousand deaths from cardiovascular disease,chronic kidney disease and stomach cancer respectively,accounting for 31.5％,30.8％ and 64.8％ of those specific causes.Comparing to the baseline in 2013,if the targets of 10％ decrease of sodium consumption by 2020 and 15％ by 2030 for Chinese chronic disease prevention and treatment planning,and 30％ decrease by 2030 for WHO non-communicable disease monitoring framework are achieved,220,340 and 730 thousand deaths will be averted,which may gain 0.30,0.45 and 0.95 years of life expectancy,respectively.Conclusions As one of the leading risk factors,diet high in sodium had caused heavy burden of disease from cardiovascular disease,chronic kidney disease and stomach cancer on Chinese residents.Intervention programs on sodium-reductionare urgently needed in China and related cost-effectiveness is highly expected.

Objective To analyze the attribution of mortality and impact on life expectancy caused by insufficient physical activity in different gender and areas in adults aged ≥25 years among Chinese people.Methods Data from the programs related to Chinese death surveillence,risk factors of chronic survey,health outcomes of physical activity as well as relative risk (RR) on Global Burden of Disease (GBD) study were used.Population attributable fraction (PAF) of different health outcomes attributable to deaths that caused physical activity and the influence of life expectancy in adults aged ≥25 years in Chinese people were calculated.Results The overall PAF for all cause of death due to physical activity in adults aged ≥25 years was 4.24％,with 4.86％ in females and 3.82％ in males.The health outcomes of inadequate physical activity would include breast cancer,colorectal cancer,ischemic heart disease,ischemic stroke and diabetes with relative PAFs as 9.04％,13.96％,14.96％,17.80％ and 16.92％,respectively.The attribution of death on Physical activity was 388 954.The most attributed death was ischemic heart disease,followed by ischemic stroke.With the elimination of physical inactivity,the total life expectancy was expected to lose by 0.43 years,with 0.47 years in women,and 0.39 years in men.Conclusion The increase of physical activity may benefit on health condition so to reduce the burden of chronic diseases and increase the life expectancy.

Objective To assess the mortality attributable to low fruit intake among people over 25 years old in China,2013,and its effect on life expectancy.Methods Based on data collected from China Chronic Disease and Risk Factor Surveillance in 2013,the average fruit intake in different genders and areas were calculated.Potential impact fraction (PIF) was used to examine the impact on deaths,mortality and life expectancy.Results The average daily fruit intake was (113.3 ± 168.9)g among people over 25 years old,with (103.6± 160.1) g for men and (122.7± 176.6) g for women,in China in 2013.Fruit intake for urban residents was significantly higher than that in rural residents and higher in eastern regions than that in central or western regions.Scores that attributable to low fruit intake accounted for 15.21％ of the total deaths and the population attributable fraction of inadequate intake of fruits to associated diseases was 35.00％.PIF for all the deaths in rural residents (16.50％)appeared higher than that of the urban residents (13.88％),and higher in the residents living in the eastern region (15.48％) than that in the central (16.27％) or western (13.75％) regions.Number of deaths that attributable to low fruit intake was 1.348 4 million.Deaths caused by related diseases appeared as:ischemic heart disease (472.5 thousands),hemorrhagic stroke (338.8 thousands),ischemic stroke (259.0 thousands),lung cancer (208.4 thousands),esophageal cancer (60.7 thousands),laryngeal cancer (5.4 thousands) and oral cancer (3.6 thousands).Numbers of all deaths and related diseases for urban residents were lower than that of the rural residents,with central regions (452.7 thousands) higher than that in the eastern (531.1 thousands) or western (364.6 thousands) regions.The average life expectancy loss caused by low fruit intake was 1.73 years,1.80 years for men and 1.58 years for women,in this country.Loss of life expectancy in the rural residents was higher than that of the urban residents,and higher in central regions than that in the eastern or western regions.Conclusions The intake of fruit was far lower than the recommended standard set for the Chinese people.Population attributable fraction was related to the associated diseases caused by inadequate intake of fruits which also made serious impact on life expectancy.

Objective:To investigate the effect of psychological support during perithrombotic period on post-stroke depression (PSD) in patients with acute ischemic stroke (AIS).Methods:Patients with AIS received intravenous thrombolysis in the Affiliated Lianyungang Hospital of Xuzhou Medical University from January 1, 2021 to July 31, 2021 were enrolled prospectively. The intervention group received one-to-one individual psychological support therapy in the perithrombolytic period on the basis of receiving standard intravenous thrombolytic therapy. At 30 d after onset, Hamilton Depression Scale was used to assess whether PSD occurred. Multivariate logistic regression analysis was used to evaluate the independent influencing factor of PSD. Results:A total of 126 patients with AIS were enrolled, and 86 of them were male (68.25%). Their age was 63.65±10.46 years; 65 were in the intervention group and 61 were in the control group. The incidence of PSD in the intervention group was significantly lower than that in the control group (20.00% vs. 36.07%; χ2=4.049, P=0.044). Multivariate logistic regression analysis showed that psychological intervention (odds ratio [ OR] 0.333, 95% confidence interval [ CI] 0.132-0.838; P=0.020] was an independent protective factor for PSD, while ischemic heart disease ( OR 4.510, 95% CI 1.181-17.217; P=0.028), alcohol consumption ( OR 3.421, 95% CI 1.317-8.888; P=0.012), anticoagulation therapy ( OR 3.145, 95% CI 1.155-8.567; P=0.025) and modified Rankin Scale score before thrombolysis ( OR 1.627, 95% CI 1.142-2.317; P=0.007) were the independent risk factors for PSD. Conclusion:Perithrombolytic psychological support may reduce the incidence of PSD.