2.Effects of sense and antisense T-STAR gene on colon cancer cell line HCT-116
Ling ZHANG ; Lian GUO ; Yong PENG ; Bin CHEN
Journal of Third Military Medical University 1988;0(06):-
Objective To investigate the effects of the sense and antisense testes-signal transduction and activator of RNA (T-STAR) gene on the colon cancer cell line HCT-116. Methods The sense and antisense T-STAR gene was stably transfected into HCT-116 cells with lipofectamine. The expression level of T-STAR in those cells was detected by Western blotting and the growth velocity and proliferation of those cells by cytokinetics. Results The growth velocity and proliferation decreased after transfection of the sense T-STAR gene, but increased after transfection of the antisense T-STAR gene. Conclusion T-STAR gene can inhibit the growth velocity and proliferation of HCT-116 cells.
3.Transfection of T-STAR antisense gene can depress the telomerase activity in lung adenocarcinoma cell line A549
Ling ZHANG ; Lian GUO ; Yong PENG ; Bin CHEN
Journal of Third Military Medical University 2003;0(11):-
Objective To investigate the changes of telomerase activity after knocking down endogenous expression of T-STAR (testes-signal transduction and activator of RNA) gene in human lung adenocarcinoma cell line A549 by antisense strategy. Methods The mRNA and protein expression of T-STAR gene were determined by RT-PCR and Western blotting, and the telomerase activity was measured by PCR-ELISA, after transfection of T-STAR antisense gene into A549 cells with lipofectamine. Sense pcDNA-STAR and blank pcDNA3.1 transfection served as control. Results The expression of T-STAR gene was significantly inhibited at mRNA and protein level, and the telomerase activity was significantly decreased. Conclusion The down-regulation of telomerase activity may result from inhibition of T-STAR gene expression in lung adenocarcinoma A549 cells.
4.Determination of 10 mycotoxin contaminants in Panax notoginseng by ultra performance liquid chromatography-tandem mass spectrometry.
Yong CHEN ; Chong-jun CHEN ; Jin LI ; Lian-jun LUAN ; Xue-song LIU ; Yong-jiang WU
Acta Pharmaceutica Sinica 2015;50(1):81-85
To ensure the quality and safety of Panax notoginseng, a method for the simultaneous determination of 10 mycotoxins in Panax notoginseng was developed using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The sample was extracted with acetonitrile and purified by HLB multifunction cleanup column. The separation was performed on a Phenomenex Kinetex XB-C18 column by gradient elution using methanol and 5 mmol·L(-1) ammonium acetate as mobile phase. The targeted compounds were detected in MRM mode by mass spectrometry with electrospray ionization (ESI) source operated in both positive and negative ionization modes. The linear relationships of the 10 mycotoxins were good in their respective linear ranges. The correlation coefficients (r) ranged from 0.9981 to 1.0000. The LOQs of the 10 mycotoxins were between 0.15 and 8.6 μg·kg(-1). The average recoveries ranged from 73.8% to 107.0% with relative standard deviations (RSDs) of 0.10%-10.9%. The results demonstrated that the proposed method was sensitive and accurate, and suitable for the mycotoxins quantification in Panax notoginseng.
Chromatography, High Pressure Liquid
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Chromatography, Liquid
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Drug Contamination
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Mycotoxins
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analysis
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Panax notoginseng
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chemistry
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Tandem Mass Spectrometry
5.Association between obesity and acute high-altitude disease
Bo YANG ; Kun LIU ; Xiaoying FENG ; Xiangmin SHI ; Chuyun MA ; Bin CHEN ; Yong XU ; Lian CHEN
Chinese Journal of Tissue Engineering Research 2007;11(34):6905-6907
BACKGROUND: The occurrence and severity of acute high-altitude disease(AHAD) are determined by the speed entering the highland, the altitude of highland and seasons. The association between obesity and AHAD has not been fully investigated.OBJECTIVE: To investigate the association between obesity and acute high-altitude in people exposed rapidly to the highland.DESIGN: Comparative observation.SETTING: Department of Cardiology, General Hospital of Chinese PLA and Department of Cardiology, Tibetan General Hospital of Chinese PLA.PARTICIPANTS: The experiment was conducted at the Department of Cardiology, General Hospital of Tibetan Military Area Command of Chinese PLA in August 2006. Totally 82 Henan subjects aged 28-45 years were selected from the male workers constructing the Qinghai-Tibet Railway and their managers with acute high-altitude exposure. They had never been to the highland before, and all agreed to the detection.METHODS: ①Every subject completed the AHAD self-report questionnaire at sea level and 12 hours and 24 hours after ascending high-altitude. The items in the questionnaire included symptoms of headache, gastrointestinal symptoms,fatigue or weakness, dizziness, and insomnia. Each symptom was graded from 0-3 with 0 as no symptoms, 1 as mild symptoms, 2 as moderate symptoms, and 3 as severe symptoms and a total score of 15. A score of 4 or more could by identified as AHAD. ②The height and body mass were measured to calculate the body mass (BMI). Those with BMI≥28 kg/m2 served as the obesity group [n =39, mean age (35±8) years], and those with BMI of 18.5-23.9 kg/m2 as the normal body mass group [n =43, mean age (35±8) years]. ③Arterial blood was taken to evaluate arterial oxygen saturation (SO2), arterial oxygen pressure (PaO2) and arterial carbon dioxide pressure (PaCO2) at baseline and 24 hours after ascending high-altitude. ④The measurement data was compared by t test.MAIN OUTCOME MEASURES: BMI, vital capacity of lungs, SO2, PaO2 and PaCO2 levels of obese and normal people.RESULTS: Totally 39 obese people and 43 normal people were involved in the result analysis. ①AHAD score: No symptom was reported at sea level in all participants (scored 0), but the AHAD scores in the obesity group were significantly higher than those in normal group 12-hour and 24-hour after ascending high-altitude. ②Blood gas analysis:At sea level, there were no statistical differences in the levels of SO2, PaO2, and PaCO2 between two groups (P > 0.05).But 24 hours after ascending high-altitude, SO2 and PaO2 of the obesity group were much lower than in the normal group (P< 0.01), and PaCO2 was significantly higher than in the normal group (P< 0.01).CONCLUSION: Obese men are more vulnerable to high-altitude hypoxia than people with normal body mass. Obesity is an important risk factor for the development of acute high-altitude disease.
6.The diagnosis value of homocysteine combined with carcinoembryonic antigen and squamous cell carcinoma antigen for esophageal cancer
Shuangping ZHANG ; Yun CHEN ; Cunzhi HAN ; Chunli WANG ; Yong MA ; Jianhong LIAN
Cancer Research and Clinic 2015;27(12):801-804
Objective To investigate the diagnosis value of homocysteine (Hcy) combined with carcinoembryonic antigen (CEA) and squamous cell carcinoma (SCC) antigen for esophageal cancer.Methods The serum levels of Hcy,CEA,CA199,CA724,CA50 and SCC antigen in 163 patients with esophageal cancer and 49 healthy people were measured.The diagnosis efficacy between the combination of Hcy,CEA and SCC antigen and the combination of CEA,CA199,CA724,CA50 and SCC antigen was compared.Results The levels of Hcy,CEA and SCC antigen were significantly raised in patients group as compared with the levels in control group (all P < 0.05).The area under the ROC curve of Hcy was 0.722 (95 % CI 0.633-0.811),CEA was 0.619 (95 % CI 0.533-0.704) and SCC antigen was 0.685 (95 % CI 0.608-0.762),respectively.There were no significant differences among the three area under the ROC curve.The sensitivity,specificity and accuracy of the combination of CEA,CA199,CA724,CA50,SCC antigen were 69.9 %,59.2 % and 67.5 %,respectively,but the sensitivity,specificity and accuracy of the combination of Hcy,CEA,SCC antigen were 96.3 %,69.4 % and 90.1%,respectively.There was significant difference between the combination of Hcy,CEA,SCC antigen and the combination of CEA,CA199,CA724,CA50,SCC antigen (P < 0.05).Conclusion The combination of Hcy,CEA,SCC antigen is helpful to determine the diagnosis and efficacy of esophageal cancer.
7.Experiences and the special medical conditions for a Chinese secondary hospital on the early stage of arterial switch operation
Kaiming CHEN ; Lang LIU ; Yong CAO ; Fenghua LAI ; Lian HU ; Bowei ZHU ; Maosheng WANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2014;30(5):269-272
Objective This retrospective study is to analysis the special medical conditions that most Chinese secondary hospitals are facing with,and to review the safeguards and pitfalls for arterial switch operations,in order to probe intothe feasibility of this procedure for Chinese secondary hospitals and provide our experiences to help other surgeons to avoid pitfalls on complex procedures.Methods Between January 2006 and December 2011,totally 21 newborns and infants with TGA/VSD and TGA/IVS underwent arterial switch operation.There were 15 males and 6 females.In the TGA/VSD group,there were 16 cases,ranging from 30 days to 1 year at surgeries,and weight from 3.4-8.5 kg with average of (5.33 ± 1.42) kg.In the TGA/IVS group,there were 5 cases,ranging from 13 days to 1 month at surgeries,and weight from 3.1-5.5 kg with average of (3.75 ± 1.17)kg.All patients underwent one stage of arterial switch operation.Routine follow-up checking points are set at discharging,3 months,half year and every year after operation.Results The early death rate is 9.5% (2/21),and the reexploration rate is 9.5% (2/21).In the TGA/VSD group,average cardiopulmonary bypass time is (151 ± 33) minuntes with the aortic crossclamp time is (1 19 ± 26) minutes.Ventilator support time is 24-159 hours,and the length of ICU stay is 3-17 days.1 case has residual VSD with the diameter less than 2 mm.The pulmonary flow velocity in 2 cases increase mildly to 2.0 m/s and 2.2 m/s,and another 2 cases increase severely to 3.1 m/s and 3.7 m/s.The aortic flow velocity in 3 cases increase to 2.0m/s.ECG instructs no case has myoinfarction signs.In the TGA/IVS group,average cardiopulmonary bypass time is (170 ± 52) minuntes with the aortic crossclamp time is (137 ± 48) minutes.Ventilator support time is 51-144 hours,and the length of ICU stay is 4-14 days;The pulmonary flow velocity in 2 cases increase mildly to 2.0 m/s.The aortic flow velocity in 1 cases increase to 2.0 m/s.ECG instructs no case has myoinfarction signs.Conclusion Unbalanced medical resources distribution causes significant differences between heart centers and the secondary hospitals in China,especially on the complex congenital heart diseases procedures.However,with relatively solid background on correction of the simple congenital heart diseases,the Chinese secondary hospitals can still perform arterial switch operation with satisfactory mortality and morbidity,and provide more prompt medical services for more population.
8.Role of the macrophages and NO in fibrosis formation in transplanted kidneys with chronic rejection
Hui-Min LIU ; Ting-Jun YE ; Yong-Lian CHEN
Academic Journal of Second Military Medical University 2001;22(4):370-372
Objective:To confirm the effects of the macrophages and NO in the fibrosis formation of transplanted kidneys with chronic rejection. Methods:The quantitive immunohistochemistry was employed to observe the changes of relative quantitiy of macrophages, NO, collagen Ⅲ andⅣ, and to reveal the association between the increases of macrophages, NO, histopathologic damages, deposition of collagen Ⅲ and Ⅳ, finally, to infer the roles of the macrophages and NO in the fibrosis formation of transplanted kidneys with chronic rejection. Results: The macrophages accumulated and increased in the glomeruli, the tubules and the intersiticia tissue of kidneys, which was companied by the increased expression of collagen Ⅲ and Ⅳ during the early and middle stages. However, the macrophages and NO could not be detected, which was companied by the decreased deposition of collagen Ⅲ and Ⅳduring the advanced stage. There were significant differences between the 3 types(glomerulus disease type,blocking vessel type and intersticial sclerostic type) (P<0.05 or P<0.01). Conclusion:The increase of macrophages and the NO may closely interrelated with the fibrosis formation in transplanted kidneys with chronic rejection.
9.Influence of cryoprotectant and cooling rate in vitrification method on the spindles of rabbit oocytes
Xue-Yong CAI ; Gui-An CHEN ; Ying LIAN ; Xiao-Ying ZHENG ; Hong-Mei PENG ;
Journal of Peking University(Health Sciences) 2003;0(06):-
SUMMARY Objective:To investigate the influence of cryoprotectants and cooling rates in vitrificationmethod on the spindles of rabbit M Ⅱ oocytes.Methods:Rabbit oocytes were verified by using cryoloopwith ethylene glycol(EG)singly or EG combined with dimethyl sulphoxide(DMSO)as cryoprotectants,and cooled by taking oocytes directly into liquid nitrogen or by vitrification machine.After frozen rabbit o-ocytes thawed,the microtubulin and chromosome of the spindles were fixation and stained by immunofluo-rescent method.Confocal microscope was used to reveal spindle configuration.Results:In the two proto-cols of single EG used and EG combined with DMSO,the spindles were severely injured.But in protocolof EG combined with DMSO and at ultra-rapid cooling rate,the normal configuration of spindle rate ofthawed rabbit oobytes was similar to that of the control group.Conclusion:The protocol of EG combinedwith DMSO as cryoprotectants and with extremely high cooling rate by vitrification machine can producethe best effect on conservation of spindle configuration in vitrification of rabbit oocytes.
10.Identification of triacylglycerols in coix oil by high performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry.
Zhi-Min XIANG ; Ming ZHU ; Bi-Lian CHEN ; Yong CHEN
China Journal of Chinese Materia Medica 2005;30(18):1436-1438
OBJECTIVETo identify triacylglycerols in coix oil.
METHODHigh performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry was used for identification. The experiment was operated under the conditions: spray voltage at 3 000 V, capillary temperature at 250 degrees C, APCI vaporizer temperature at 400 degrees C, and corona current of 4 microA. Sheath gas pressure (high purity liquid nitrogen) was 35 kPa. Mass spectra were obtained over the m/e range of 300 to 900 amu, scan duration of 1s and Q1 peak width at 0.7. The stationary phase was Zorbax Extend C18 column (4.6 mm x 250 mm, 5 microm). The mobile phase: dichloromethane-acetonitrile (35:65), flow rate: 1 mL x min(-1); column temperature: 25 degrees C.
RESULT12 triacylglycerols were identified by HPLC-MS method.
CONCLUSIONThe result can be used to identify the components in a fingerprint chromatogram of coix oil and its related injection product.
Chromatography, High Pressure Liquid ; methods ; Coix ; chemistry ; Mass Spectrometry ; methods ; Plant Oils ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Seeds ; chemistry ; Triglycerides ; analysis ; chemistry ; isolation & purification