1.Purification of large-scale plasmid DNAs by selective precipitation with cetyltrimethylammonium bromide.
Quan ZHANG ; Kexiang YU ; Weifeng YUAN ; Fangming XUE ; Huaichang SUN ; Hongfei ZHU
Chinese Journal of Biotechnology 2008;24(12):2117-2121
Following Escherichia coli lysis with alkali, cetyltrimethylammonium bromide (CTAB) was directly titrated into the supernatant. An easy and feasible technology for plasmid purification was established with the optimized proportion between the quantity of CTAB and plasmid, combined with the specific solution for DNA release and TritonX-114 for endotoxin removal. Quality detection showed that the purified plasmid was free of contamination of host RNA. The host bacterial genomic DNA, endotoxin and bacterial protein were less than 10 microg, 50 EU and 10 microg per mg plasmids, respectively. The ratio of OD260/OD280 was between 1.75-1.85. Eighty percent of the prepared plasmids were presented in the supercoiled form. The plasmid purified with this technology can satisfy all criteria stipulated by FDA. The main advantages of the technology include the avoidance of animal-derived enzymes such as ribonucleases A, Proteinase K and toxic reagents like chloroform and phenol. In addition, the technology has low cost and no pollution.
Cetrimonium Compounds
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chemistry
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Chemical Precipitation
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DNA
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genetics
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isolation & purification
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DNA, Bacterial
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genetics
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isolation & purification
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Escherichia coli
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chemistry
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genetics
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Plasmids
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genetics
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isolation & purification
2.The relationship between chronic obstructive pulmonary disease and bone mineral density in Chinese adults
Kexiang SHI ; Yunqing ZHU ; Canqing YU ; Jun LYU ; Yu GUO ; Pei PEI ; Ling YANG ; Huaidong DU ; Yiping CHEN ; Junshi CHEN ; Zhengming CHEN ; Liming LI
Chinese Journal of Epidemiology 2023;44(2):221-228
Objective:To investigate the relationship between chronic obstructive pulmonary disease (COPD) and bone mineral density (BMD) in Chinese male and female adults.Methods:Linear regression models were used to analyze the correlations between COPD and the Global Initiative for Chronic Obstructive Lung Disease (GOLD) grade concerning bone mineral density indices, including broadband ultrasound attenuation, speed of sound, and stiffness index, based on data from the second resurvey of China Kadoorie Biobank.Results:Among 23 876 participants, the prevalence of COPD was 34.3% (3 130/9 125) in males and 23.2% (3 416/14 751) in females. Compared with healthy males and healthy females, broadband ultrasound attenuation, speed of sound, and stiffness index decreased in males and females with COPD, with β values (95% CI) of -0.87 (-1.37- -0.36), -3.42 (-5.42- -1.43) and -1.53 (-2.34- -0.71) in males and -0.66 (-1.09- -0.23), -2.24 (-3.92- -0.55), -1.06 (-1.71- -0.40) in females, respectively. The decrease was greater in males than females (all P for interaction <0.05). The GOLD grade of COPD was inversely correlated with all the three indices in a dose-response manner (all P for trend <0.001). Subgroup analysis showed that patients with COPD had a greater decrease in BMD in males whose ages were ≥ 60 years, males who were less physically active, and participants who were not overweight or obese. Conclusions:COPD was negatively correlated with bone mineral density. More attention should be paid to the bone mineral density of patients with COPD, especially those with older age, less physical activity, or lower BMI.
3.Discussion on the influencing factors of beating heart coronary artery bypass grafting
LIU Yun ; PIAO Hulin ; LI Bo ; WANG Yong ; XIE Chulong ; WEI Shibo ; XU Jian ; GAO Xinghao ; DU Yu ; LIU Kexiang
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2019;26(1):73-77
Objective To explore the factors affecting the operation of coronary artery bypass grafting with heart beating and improve the effect of the operation. Methods From January 2012 to June 2016, 898 patients with coronary heart disease who received cardiovascular surgery in the Second Affiliated Hospital of Jilin University were analyzed retrospectively. All patients only underwent coronary artery bypass grafting with beating heart. Among them, 797 patients underwent the off-pump coronary artery bypass grafting (an OPCABG group, 592 males and 205 females, with an average age of 60.5±8.4 years); another 101 patients received on-pump beating heart coronary artery bypass grafting (an OPBH group, 77 males and 24 females, with an average age of 61.5±8.2 years). Results The average number of grafts in the OPCABG group was 3.36±0.74, and in the OPBH group was 3.71±0.69 (P<0.05). The postoperative ventilation time (10.8±9.5 hvs. 20.6±12.3 h), ICU stay (28.8±15.5 h vs. 37.4±30.8 h), hospital stay (10.9±4.8 d vs. 14.8±8.6 d), mortality (1.1% vs. 3.0%), the utilization rate of intra-aortic balloon pump (2.4% vs. 8.9%) and extracorporeal membrane oxygenation (0.5% vs. 5.0%) were significantly different between the OPCABG group and OPBH group (all P<0.05). Twelve patients died after surgery, and the total bloodless operation ratio was 91.3%. Conclusion The results show that most patients can achieve good results with the help of apical fixation and myocardial fixator, improved surgical techniques and methods, good anesthesia management as well as flexible and accurate use of vasoactive drugs. But extracorporeal circulation is necessary in the patients with large left ventricle, low ejection fraction and hemodynamic instability after intraoperatively moving the heart.
4.Prevalence of psoriatic arthritis in Chinese population with psoriasis: A multicenter study conducted by experienced rheumatologists.
Hua ZHANG ; Miao CHEN ; Ran CUI ; Xia LI ; Kexiang YAN ; Lihong CHEN ; Zhenghua ZHANG ; Ning YU ; Xinling BI ; Hui DENG ; Yangfeng DING ; Qiong HUANG ; Sheng-Ming DAI
Chinese Medical Journal 2023;136(12):1439-1447
BACKGROUND:
Reports on the prevalence of psoriatic arthritis (PsA) among Chinese patients with psoriasis are very limited. This study, conducted by rheumatologists, estimated the prevalence of PsA in a large number of Chinese patients with psoriasis.
METHODS:
Consecutive patients with a confirmed diagnosis of psoriasis attending nine dermatology clinics in five hospitals were recruited. All psoriasis patients were asked to complete a questionnaire comprising 16 questions to identify possible cases of PsA. All patients with one or more positive answers to the questionnaire were evaluated by two experienced rheumatologists.
RESULTS:
A total of 2434 psoriasis patients, including 1561 males and 873 females, were enrolled. Both the questionnaire and rheumatologists' examinations were completed in the dermatology clinics. The results identified 252 patients with PsA, comprising 168 males and 84 females. The overall prevalence of PsA among psoriasis patients was 10.4% (95% confidence interval [95% CI], 9.1%-11.7%). By sex, the prevalence was 10.8% (95% CI, 9.2%-12.5%) for males and 9.6% (95% CI, 7.7%-11.9%) for females and there was no significant sex difference in the prevalence of PsA (P = 0.38). Of the 252 PsA patients, 125 (49.6%, 95% CI, 41.3%-59.1%) were newly diagnosed by rheumatologists. Consequently, the prevalence of undiagnosed PsA among psoriasis patients was 5.2% (95% CI, 4.4%-6.2%).
CONCLUSION
The prevalence of PsA in the Chinese population with psoriasis is about 10.4%, which is almost double that of previous reports in the Chinese population, but lower than that in Caucasians.
Humans
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Female
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Male
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Arthritis, Psoriatic/epidemiology*
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Rheumatologists
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Prevalence
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East Asian People
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Psoriasis/epidemiology*