1.Protective Effect of Angiotensin Converting Enzyme 2 Transfection on Vascular Endothelial Cells
Qingqing HAO ; Yonghuan ZHANG ; Qingtao YU ; Li ZHU ; Xu CHEN ; Shuying LI ; Yuehui ZHANG ; Bo DONG ; Ruifeng LI
Chinese Circulation Journal 2014;(10):841-845
Objective: To investigate the inlfuence of angiotensin converting enzyme 2 (ACE2) on lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) protein expression and to explore the protective effect of ACE2 on vascular endothelial cells. Methods: Our work includedin vitro andin vivo studies. For in vitro experiment, the human umbilical vein endothelial cell (HUVEC) were cultured and transfected with replication deficient recombinant adenovirus of ACE2 (Ad-ACE2), and LOX-1 protein expression stimulated by angiotensin 2 was examined by Western blot analysis. Forin vivo study, atherosclerosis plaques were induced in 20 apolipoprotein E-deifcient (ApoE-/-) mice, and then randomly divided them into 2 groups: ACE2 group, the mice received Ad-ACE2 (2.5×109 pfu/ml) injection through caudal vein, EGFP (enhanced green lfuorescent protein) group, the mice received equal replication deifcient recombinant adenovirus of EGFP (Ad-EGFP) injection through caudal vein.n=10 in each group. The animals were executed after 1 month treatment to collect abdominal aorta. Lipid content in atherosclerosis plaque was evaluated by Oil red O staining and LOX-1 protein expression was examined by immunohistochemistry and Western blot analysis. Results: Bothin vitro andin vivo experiments conifrmed that endothelial cell LOX-1 protein expression was signiifcantly inhibited by ACE2 transfection. The lipid content in ACE2 group was obviously lower than that in EGFP group byin vivo study. Conclusion: ACE2 may inhibit LOX-1 protein expression and therefore reduce the progress of atherosclerosis.
2.Application of HTB-SiHa Cells Transfected with a Recombinant Plasmid for External Quality Assessment of Chlamydia trachomatis PCR.
Kuo ZHANG ; Hong HUO ; Yu SUN ; Lunan WANG ; Rui ZHANG ; Guigao LIN ; Jiehong XIE ; Qingtao WANG ; Jinming LI
Annals of Laboratory Medicine 2014;34(5):360-366
BACKGROUND: The participation of laboratories in external quality assessment (EQA) programs is required for the quality assurance of nucleic acid amplification of Chlamydia trachomatis. This study aimed to construct a new quality control (QC) material applicated in EQA of C. trachomatis PCR. METHODS: A QC material-HTB-SiHa cells transfected with a recombinant plasmid containing the cryptic plasmid sequence-was constructed for C. trachomatis PCR detection, and four different panels, each consisting of 4 positive samples with serial dilution of the constructed QC material and 1 negative sample, were distributed by the National Center for Clinical Laboratories among four groups of 275, 268, 317, and 304 participants across China from 2011 through 2012. A total of eight commercial kits were used for C. trachomatis PCR detection in participants. RESULTS: Nine laboratories reported false-positive results (0.9%). As the series dilution increased, the correct reporting of the data sets decreased; the lowest correct rate was 96.3% in the weakest positive samples (104 copies/mL). Eight laboratories reported false-positive results, and 42 laboratories reported false-negative results in the EQA detection of C. trachomatis. No significant differences were observed in the detection of the constructed C. trachomatis positive samples (97.9%, 98.5%, 100%, 98.5%; P=0.36) and negative samples (100%, 99.0%, 100%, 99.0%; P=0.764) using four commercial kits commonly used in China. CONCLUSIONS: The results of the EQA study indicated that the constructed material provides a noninfectious, stable control material with sufficient volume for PCR detection of C. trachomatis.
Cell Line
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Chlamydia Infections/diagnosis
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Chlamydia trachomatis/*genetics
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DNA, Bacterial/*analysis
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False Negative Reactions
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Humans
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Laboratories/*standards
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Plasmids/genetics/*metabolism
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Polymerase Chain Reaction/*standards
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Quality Control
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Reagent Kits, Diagnostic
3.Association of insulin resistance with common carotid arterial stiffness in hemodialysis patients
Yilun ZHOU ; Zexing YU ; Huimin JIA ; Qingtao WANG ; Juan MENG ; Fang SUN ; Lijie MA ; Jing LIU ; Bin HAN ; Qianmei SUN ; Liren PENG
Chinese Journal of Nephrology 2008;24(4):249-252
Objective To investigate the association between arterial stiffness of the common carotid artery(CCA)and insulin resistance in hemodialysis patients. Methods Arterial stiffness index β of CCA was evaluated by an ultrasonic phase-lock Echo-tracking system in 80stable non-diabetic hemodialysis patients.Insulin resistance was detected by the homeostasis model assessment method(HOMA-IR).Plasma hemoglobin,serum albumin,total cholesterol,high density lipoprotein,low density lipoprotein,triglyceride,lipoprotein(a),ApoA1,ApoB,CRP,calcium,phosphorus and creatinine were determined by standard methods. Results The stiffness index β was 11.41±4.13 in patients with previous cardiovascular disease(CVD)and 9.75±3.63 in those without CVD(P<0.05).The stiffness index β was positively correlated with HOMA-IR(r=0.321,P<0.01),as well as with age(r=0.376,P<0.01),pulse pressure(r=0.267,P<0.05),and duration of hemodialysis(r=0.219,P<0.05).In stepwise multiple regression analysis,HOMA-IR(β=0.228,P<0.05)and age(β=0.308,P<0.01)were identified as significant independent variables for stiffness index β of CCA. Conclusions Insulin resistance is associated with aaefial stiffness in nondiabetic hemodialysis patients.The increased arterial stiffness may be the link between insulin resistance and cardiovascular morbidity as well as mortality in hemodialysis patients.
4.Pterostilbene alleviates renal ischemia-reperfusion injury in mice by activating mitochondrial autophagy
Hao MING ; Yunyan ZHANG ; Shenglan MEI ; Yanli YU ; Qingtao MENG ; Zhongyuan XIA
Chongqing Medicine 2023;52(23):3533-3537
Objective To study the protective effect and related mechanism of pterostilbene(PTE)on renal ischemia-reperfusion injury(IRI)in mice.Methods Twenty-four C57 mice were randomly divided into 4 groups(n=6),including the sham operation group(S group),the renal ischemia reperfusion group(IR group),the renal ischemia reperfusion+5 mg/kg PTE group(IR+PTE1 group)and the renal ischemia reper-fusion+10 mg/kg PTE group(IR+PTE2 group).Hematoxylin-eosin(HE)and TUNEL staining were used to evaluate renal tissue injury.Creatinine(Cr),blood urea nitrogen(BUN),malondialdehyde(MDA)in renal tissue,inflammatory cytokines tumor necrosis factor(TNF-α),interleukin(IL)-1β and IL-6 levels in serum were detected with relevant kits.The expression of Bcl-2 interacting protein 3(BNIP3)and microtubule-asso-ciated protein light chain(LC)-3 Ⅱ in kidney tissue was detected by Western blot.Results Compared with the S group,serum levels of Cr,BUN,MDA,TNF-α,IL-1β and IL-6 in the IR group were increased,renal tu-bule injury score and apoptosis index were increased,and expressions of BNIP3 and LC-3 Ⅱ were down-regula-ted,with statistical significance(P<0.05).Compared with the IR group,serum levels of Cr,BUN,MDA,TNF-α,IL-1β and IL-6 in the IR+PTE1 group were decreased,renal tubule injury score and apoptosis index were decreased,and expressions of BNIP3 and LC-3 Ⅱ were up-regulated,with statistical significance(P<0.05).Compared with the IR+PTE1 group,serum levels of Cr,BUN,MDA,TNF-α,IL-1β and IL-6 in the IR+PTE2 group were decreased,renal tubule injury score and apoptosis index were decreased,and expres-sions of BNIP3 and LC-3 Ⅱ were up-regulated,with statistical significance(P<0.05).Conclusion PTE has protective effect on renal IRI in mice.
5.Comparison of three oral regimens with compound polyethylene glycol electrolyte for bowel prepara-tion
Jinfang ZHAO ; Liangru ZHU ; Hongyu REN ; Jun LIU ; Xiaohua HOU ; Jie WU ; Shengbin SUN ; Yijuan DING ; Shiyun TAN ; Xiaohong LU ; Meifang HUANG ; Jin LI ; Min CHEN ; Zili DAN ; Peiyuan LI ; Wei YAN ; Qingtao MEI ; Weizhong YU
Chinese Journal of Digestive Endoscopy 2015;(9):613-616
were no significant differences in the detection rate of recto-sigmoid colon,mid colon,right colon and total detection of polyps among the 3 groups (P >0.05).Conclusion 4-L split-dose PEG is better than the oth-er 2 regimens in the colon cleansing quality,so it can better reach the intestinal cleaning standards before enteroscopy,which is a more suitable regimen for bowel preparation.
6.Detection of a BRCA1 c.2013_2014ins GT variant an ethnic Han Chinese pedigree affected with breast cancer.
Pan QI ; Linlin GAO ; Xiaoying HE ; Yuehan NI ; Sheng XU ; Xueying MAI ; Guiling ZHANG ; Yuxia LIU ; Yu GUO ; Yong ZHOU ; Qingtao HU
Chinese Journal of Medical Genetics 2020;37(4):415-418
OBJECTIVE:
To detect potential variant in an ethical Han Chinese pedigree affected with breast cancer.
METHODS:
The proband and her relatives were subjected to next-generation sequencing using a target capture sequencing kit containing 121 cancer-related genes. Candidate variants were selected by analysis of their type, frequency in population, and segregation with the phenotype. Candidate variant was verified by Sanger sequencing and TA cloning.
RESULTS:
A c.2013_2014ins GT variant was detected in the BRCA1 gene among all breast cancer patients from this pedigree but not among healthy females. The variant was not recorded in the 1000 Genome Project database or the Exome Aggregation Consortium (ExAC) database. The frameshifting insertion was predicted to form an premature stop codon in gene transcript and can give rise to a truncated protein.
CONCLUSION
The BRCA1 c.2013_2014ins GT variant probably underlies the pathogenesis of breast cancer in this Chinese pedigree.
Asian Continental Ancestry Group
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BRCA1 Protein
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genetics
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Breast Neoplasms
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genetics
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Exome
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Female
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High-Throughput Nucleotide Sequencing
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Humans
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Pedigree
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Phenotype
7.Reflection on the standardized training of geriatric specialists under the background of "the aging of population"
Qingtao HOU ; Weihua YU ; Caishuang PANG ; Li XIA ; Qian XIAO ; Lü YANG
Chinese Journal of Medical Education Research 2023;22(7):1069-1073
With the intensive aging of the population, it's imperative and important to train a number of geriatric specialists. Essential clinical knowledge and skills as well as accomplishment of medical humanistic spirit are core competences of an eligible geriatrician. The standardized training of geriatric specialists is facing a few challenges such as incomplete comprehension of the training program and trainees, a lack of enough trainees, and a lack of standardized management for the program. An efficient social support system, a normative educational training system, an effective supervision and evaluation system, first-class teaching staff, and qualified trainees are important guarantees for the standardized training program.
8.Establishment of reference intervals for thyroid stimulating hormone measurement by big data and indirect method in adults
Shunli ZHANG ; Yu MO ; Fei CHENG ; Tingting JIA ; Yue ZHAO ; Mo WANG ; Yuhong YUE ; Rui ZHANG ; Jing XU ; Yang ZHAO ; Qingtao WANG
Chinese Journal of Laboratory Medicine 2021;44(7):627-632
Objective:To establish the sex-, age-and season-specific (month) reference intervals (RI) for thyroid stimulating hormone (TSH) measurement by big data and indirect method in adults.Methods:TSH data of anonymous patients were collected from Beijing Chaoyang Hospital Affiliated to Capital Medical University in 2016, the data were selected and outliers were removed. Indirect methods (Hoffmann method and Bhattacharya method) were used to calculate TSH reference intervals of whole population, different genders, ages and seasons (months). TSH RI from two indirect methods of total population, selected population, physical examination population was compared with RI from reagent instruction according to reference change value ( RCV) based on biological variability. Results:A total of 61 599 records were obtained from 90 699 records including 18 776 males and 42 823 females. The TSH RI were obtained by Hoffmann method: the whole population, 0.59-5.59 μIU/ml (1 μIU/ml=1 mIU/L), male, 0.53-5.16 μIU/ml, female, 0.59-6.11 μIU/ml. The upper limits of TSH RI were higher with age and in winter (January): 18-30 years old, 0.62-5.57 μIU/ml, 71-80 years old, 0.49-6.45 μIU/ml; January, 0.59-6.40 μIU/ml, August, 0.60-5.56 μIU/ml; The RI of TSH by Bhattacharya method: the whole population, 0.58-5.80 μIU/ml, male, 0.55-5.02 μIU/ml, female, 0.62-6.21 μIU/ml. The upper limits of TSH RI were also higher with age and in winter (January): 18-30 years old, 0.65-5.67 μIU/ml, 71-80 years old, 0.46-5.99 μIU/ml, January: 0.61-6.52 μIU/ml, August: 0.61-5.69 μIU/ml. Compared to RI from reagent instruction, the differences of TSH RI from two indirect methods of total population, selected population, physical examination population were acceptable.Conclusions:TSH RI was established by indirect method. With the increase of age and winter, the upper limit of TSH reference interval tends to increase.
9.Recent developments in L-lactate fermentation by genetically modified microorganisms.
Xu JIANG ; Limin WANG ; Guimin ZHANG ; Bo YU ; Qingtao ZENG
Chinese Journal of Biotechnology 2013;29(10):1398-1410
Lactic acid is an important platform chemical. Especially with rapid development of poly (lactic acid) industry, the demand for L-lactic acid is continuously increasing. To further reduce the fermentation costs and improve the robustness of strains from industrial point of view, many modern biotechnological approaches have been applied to strain development. In this review, we briefly summarize recent advances in L-lactate fermentation by genetically modified microorganisms, including lactic acid bacteria, yeast, E. coli and Rhizopus species.
Bacteria
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genetics
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metabolism
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Escherichia coli
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genetics
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metabolism
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Fermentation
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Genetic Engineering
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methods
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Industrial Microbiology
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methods
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Lactic Acid
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metabolism
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Metabolic Networks and Pathways
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genetics
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Stereoisomerism
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Yeasts
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genetics
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metabolism
10.Content Determination of 8 Components in Astragalus membranaceus before and after Bidirectional Solid Fer- mentation with Cordyceps kyushuensis by HPLC-DAD-ELSD
Yiqing DU ; Qinhui YU ; Ran WEN ; Tongtong JIAN ; Rong RONG ; Qingtao LYU ; Guoying ZHANG
China Pharmacy 2020;31(23):2841-2846
OBJECTIVE:To establish a method for simultaneous determination of calycosin glucoside ,ononin,calycosin, formononetin,astragaloside Ⅳ,isoastragaloside Ⅱ,cycloastragenol and isoastragaloside Ⅰ in Astragalus membranaceus before and after bidirectional solid fermentation with Cordyceps kyushuensis ,and to investigate the effects of fermentation on the contents of above 8 components in A. membranaceus . METHODS :HPLC-DAD-ELSD was adopted. The determination was performed on Agilent 5 TC-C18 column with mobile phase consisted of 0.1% formic acid aqueous solution-acetonitrile (gradient elution )at the flow rate of 1 mL/min. The column temperature was set at 30 ℃. DAD detection wavelength was set at 260 nm,ELSD evaporation tube temperature was 100 ℃,atomizer temperature was 80 ℃,carrier gas flow rate was 1.6 L/min;injection volume was 15 μL. RESULTS:The eight components had a good linear relationship within their respective ranges of concentration (all R2>0.999 0); RSDs of precision ,stability and repeatability tests were all less than 3%(n=3 or n=6);the recoveries was 97.88%-101.32%, and RSDs were 1.22%-2.39%(n=6). Setting the content of components in unfermented A. membranaceus as 100%,after bidirectional solid fermentation with C. kyushuensis ,the change rates of 8 components were -98.51%,-96.41%,-94.74%, -96.40%,289.20%,20.25%,-75.05%,562.46%,respectively. CONCLUSIONS :After fermentation with C. kyushuensis ,the contents of active components as astragaloside Ⅳ,isoastragaloside Ⅰ and isoastragaloside Ⅱ can be increased significantly in A. membranaceus .