2.Chlorpyrifos Determined in Human Blood by UPLC-MS/MS and Its Application in Poisoning Cases.
Zheng QIAO ; Hui YAN ; Xian-yi ZHUO ; Bao-hua SHEN
Journal of Forensic Medicine 2015;31(2):112-116
OBJECTIVE:
To determine the chlorpyrifos in human blood by liquid chromatography-tandem mass spectrometry and to validate its application in poisoning cases.
METHODS:
The samples were extracted by a simple one-step protein precipitation procedure. Chromatography was performed on a Capcell Pack C18 MGII column (250 mm x 2.0 mm, 5 μm) using an isocratic elution of solvent A (0.1% formic acid-water with 2 mmol/L ammonium acetate) and solvent B (methanol with 2 mmol/L ammonium acetate) at 5:95 V:V).
RESULTS:
The linear ranged from 5 to 500 ng/mL (r = 0.998 7). The limit of detection (LOD) and the lower limit of quantification (LLOQ) were 2 ng/mL and 4 ng/mL, respectively. For this method, the precision and accuracy of intra-day and inter-day were < 10% and 97.44%-101.10%, respectively. The results in stability test of long-term frozen were satisfied. The matrix effect, recovery and process efficiency were 64.97%-86.81%, 76.70%-85.52%, and 55.57%-66.58%, respectively.
CONCLUSION
This method can provide a rapid approach to chlorpyrifos extraction and determination in toxicological analysis of forensic and clinical treatment.
Chlorpyrifos/blood*
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Chromatography, High Pressure Liquid/methods*
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Chromatography, Liquid/methods*
;
Humans
;
Limit of Detection
;
Poisoning
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Reproducibility of Results
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Tandem Mass Spectrometry/methods*
3.Study on the Optimum Culture for Growth of Bacillus Subtilis B115 andthe Antibacterial Effect of B115 on Aeromonas
Zhi-Hua SHEN ; Jin-Yu SHEN ; Wen-Lin YIN ; Xiao-Yi PAN ; Ying-Lei WU ;
Microbiology 1992;0(04):-
The essential medium of B115 composed of 1% tryptone, 0.25% yeast extract and 0.5% sodium chloride was determined by using an orthogonal design. The orthogonal design was also employed in testing the optimum additions. It was composed of 0.1%(NH_(4))_(2)SO_(4),1.4%K_(2)HPO_(4), 0.6% KH_(2)PO_(4) and 0.1% (Na_(3)C_(6)H_(5)O_(7)). The yield of B115 cultured in optimum medium was compared with the one in essential medium. Statistic analysis showed that the growth of B115 was most significantly improved by adding K~(+)、NH~+_(4) and (Na_(3)C_(6)H_(5)O_(7)) to essential medium. The antibacterial effect of Bacillus subtilis strain B115 on pathogenic Aeromonas was studied. The results showed different antibacterial effects of B115 on different aeromonads. There were obvious antibacterial effects on BSK-10 and CL990920, while no effect on the growth of TL970424.
4.Screening of benzodizepines and their metabolites in urine.
Bao-hua SHEN ; Min SHEN ; Xian-yi ZHUO
Journal of Forensic Medicine 2002;18(1):22-25
OBJECTIVE:
Screen seventeen benzodizepines and their metabolitesin urine by GC/ECD and GC/MS.
METHODS:
They were GC (GC/ECD, GC/MS) assay of benzodizepines and GC (GC/ECD, GC/MS) assay of benzophenones of acid-hydrolytic products of 1,4-benzodizepines.
RESULTS:
The methods were simple and sensitive. The recoverys were 60% to 90% of most benzodizepines, linear calibration curves were 20 ng/ml-200 ng/ml (r > 0.99), and detection limits were 0.5 ng/ml-10 ng/ml.
CONCLUSION
They methods were evaluated with human urine samples.
Anti-Anxiety Agents/urine*
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Benzodiazepines/urine*
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Benzophenones/urine*
;
Chromatography, Gas
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Gas Chromatography-Mass Spectrometry
;
Humans
5.Dynamic changes of regulatory T cells and their inhibitory molecules during antiviral treatment in patients with chronic hepatitis C
Zhiqin LI ; Xinyu GU ; Yu PING ; Jinxing HU ; Hua LI ; Jingya YAN ; Shen SHEN ; Zujiang YU ; Yi ZHANG
Chinese Journal of Infectious Diseases 2016;34(6):332-336
Objective To investigate the dynamic changes of regulatory T cells (Treg ) and the surface expression of programmed death (PD)‐1 and the level of transforming growth factor (TGF )‐βduring antiviral treatment in patients with chronic hepatitis C (CHC) .Methods Eighty‐six CHC patients referred to the First Affiliated Hospital of Zhengzhou University from October 2012 to October 2013 were included ,and all of them were administered with pegylated interferon α‐2a and ribavirin .Thirty healthy controls were enrolled .The percentage of Treg cells ,PD‐1 expression and TGF‐β level were analyzed by flow cytometry at baseline and at time of achieving rapid virological response (RVR ) , early viral virological (EVR ) , end‐of‐treatment virological response (ETVR ) and sustained virological response (SVR) ,or not achieving SVR .Comparison between two groups was analyzed by t test .Results Among 86 CHC patients ,the proportions of RVR ,EVR ,ETVR ,and SVR at week 24 of follow‐up were 29 cases ,67 cases ,79 cases and 67 cases ,respectively .Percentage of Treg cells in CHC patients was much higher than that in healthy controls (10 .31 ± 5 .61 vs 2 .18 ± 0 .65 ,t = 2 .28 , P< 0 .05) .During antiviral therapy ,percentages of Treg cells declined ,not only in CHC patients with HCV genotype 1b (at baseline , RVR ,EVR ,and ETVR :14 .44 ± 3 .78 ,11 .01 ± 1 .79 ,8 .24 ± 2 .98 ,and 5 .36 ± 1 .47 ,respectively ) ,but also in those infected with HCV genotype 2a (at baseline ,RVR ,EVR ,and ETVR :12 .34 ± 2 .82 ,8 .99 ± 1 .68 ,7 .53 ± 2 .96 ,and 4 .79 ± 1 .23 ,respectively ) .Expressions of PD‐1 and TGF‐β also decreased .At baseline ,the expressions of PD‐1 in patients with SVR and without SVR were 29 .11 ± 14 .65 and 37 .73 ± 11 .65 ,respectively (t = 2 .15 , P = 0 .04) ,and the levels of TGF‐β were 41 .20 ± 18 .96 and 56 .75 ± 14 .42 ,respectively (t= 2 .66 ,P< 0 .01) .At week 24 ,the expressions of PD‐1 in patients with SVR and without SVR were 10 .36 ± 4 .81 and 36 .46 ± 10 .52 ,respectively (t= 13 .95 ,P< 0 .01) ,and the levels of TGF‐β were 10 .06 ± 4 .64 and 45 .23 ± 17 .85 , respectively ( t = 11 .85 , P < 0 .01 ) . Conclusions Percentages of Treg cells and expressions of PD‐1 and TGF‐β decrease during antiviral treatment in CHC patients .Thus ,it could be of assist to predict the treatment response by monitoring these parameters .
7.THE THERAPEUTIC EFFECTS OF A HIGH FIBRE DIET ON THE EXPERIMENTAL DIABETES AND CLINICAL DIABETES MELLITUS
Mingzhi XIE ; Guoxian SHAO ; Zhufang SHEN ; Yi HUA ; Qiming CHEN ; Haifan LIU
Acta Nutrimenta Sinica 1956;0(02):-
The therapeutic effects of a high fibre diet composed of naturally high fibre foodstuffs containing protein, various essential amino acids and trace elements (Zn, Cr etc.) on diabetics were tested both experimentally and clinically. The high fibre diet or steamed bread (for control) with identical carbohydrate content was taken by normal mice, alloxan diabetic mice and nine healthy volunteers. The high fibre diet caused significant reduction in the blood glucose area (carbohydrate tolerance) as compared with the steamed bread. In another study, eighteen non-insulin dependent diabetic patients (10 females, 8 males) were administered the high fibre diet for 34 days, the average fasting and 2 h postprandial blood glucose level were significantly lower than those before the test, but no difference was observed in blood electrolytes. This implied that no malabsorption occurred during the testing period. At the same time, 16 of the 17 overweight patients were found to have a reduction of weights. In addition, improvement of symptoms (poly-dipsia polyuria, constipation etc) both in diabetic animals and in patients was observed. This study indicates that the high fibre diet is benificial for diabetic patients.
8.Expression and significance of hypoxia-inducible factor 1alpha protein and microvessel density marked by CD105 in cervical carcinoma.
Yan ZHU ; Qi-chang YANG ; Hong-bin LIU ; Man-hua LIU ; Yi SHEN ; Xiao-juan ZHANG
Chinese Journal of Pathology 2008;37(4):264-265
Adult
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Aged
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Aged, 80 and over
;
Antigens, CD
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immunology
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Endoglin
;
Female
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Gene Expression Regulation, Neoplastic
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Humans
;
Hypoxia-Inducible Factor 1, alpha Subunit
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genetics
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metabolism
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Microvessels
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immunology
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Middle Aged
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Nitric Oxide Synthase Type II
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metabolism
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Receptors, Cell Surface
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immunology
;
Uterine Cervical Neoplasms
;
genetics
;
metabolism
9.OPTIMIZATION OF THREE-CUBE FRET ASSAY AND APPLICATION IN THE ANALYSIS OF SUBUNIT ASSEMBLING OF iGluR RECEPTORS IN LIVING CELLS
Minzhen WANG ; Shuang QIU ; Yulin HUA ; Fan YANG ; Yi SHEN ; Jianhong LUO
Acta Anatomica Sinica 1957;0(04):-
Objective To establish and optimize three-cube FRET assay in living cells and analyze subunit assembling of iGluR receptors. Methods Taking HEK293 cells cotransfed with pECFP and pEYFP as negative control, and those transfected with pECFP-YFP as positive control,different calculation methods using fluorescence microscopy were compared. Results These calculation methods were all suitable for FRET measurement in the system. but the measurement results were affected by the ratio of Donor/Acceptor (D/A) in some degree,and different calculation methods have different optimized conditions. FRET measurement using FR value showed subunit specific assembly of iGluR subtypes.Conclusion There are different optimized conditions for these different calculation methods in the three-cube FRET measurement system,and a further evidence is provided for subunit specific assembling of iGluR subtypes from the FRET assay.
10.Expression and roles of long non-coding RNA Linc00467 in lung adenocarcinoma
Zhuangzhuang CONG ; Zhong GUO ; Tao QIN ; Yong QIANG ; Hua JING ; Yi SHEN
Journal of Medical Postgraduates 2017;30(8):834-838
Objective The role of long non-coding RNA Linc00467 in human lung adenocarcinoma is not yet clear.This study was to investigate the expression of long non-coding RNA Linc00467 in human lung adenocarcinoma, its clinical significance, and the effects of Linc00467 on the functions of the tumor and endothelial cells in vitro.Methods Lung adenocarcinoma tissue and normal tissue surrounding the malignance were obtained from 60 patients with pathologically proved stage I-Ⅲa lung adenocarcinoma.Human umbilical vein endothelial cells (HUVECs) were transfected with the over-expressed plasmid pccl-Linc00467 (HUVEC experimental group) or the empty vector pccl (HUVEC control group), A549 cells with Linc00467-siRNA (A549 experimental group) or negative siRNA (A549 control group), and H1299 cells, too, with Linc00467-siRNA (H1299 experimental group) or negative siRNA (H1299 control group).The expression level of Linc00467 in the lung adenocarcinoma tissue was detected by qRT-PCR with an analysis of its correlation with the clinicopathological characteristics of the patients;the influence of Linc00467 on the proliferation of the A549, H1299 and HUVEC cells was assayed with CCK-8;and the role of Linc00467 in the angiogenesis of the HUVECs was assessed by fibrin bead sprouting assay.Results The expression of Linc00467 in the lung adenocarcinoma tissue was 2.72±1.31 times as high as that in the normal lung tissue (P<0.01), and those in the A549 and H1299 cells were 3.45±0.25 and 3.22±0.33 times as high as those in the human bronchial epithelial (HBE) cells (P<0.01).The expression level of Linc00467 was significantly correlated with the tumor size and vascular invasion (P<0.05).After transfection of Linc00467-siRNA, the expressions of Linc00467 in the A549 and H1299 experimental groups were down-regulated by 72% and 68% as compared with those in the A549 and H1299 control groups (P<0.01).The number of living cells was remarkably decreased in the A549 experimental group in comparison with the A549 control at 48 h (1.29±0.07 vs 1.51±0.09), 72 h (1.53±0.15 vs 2.13±0.11), and 96 h after culturing (1.98±0.18 vs 3.02±0.12), and so was it in the H1299 experimental versus the H1299 control group, but markedly increased in the HUVEC experimental versus the HUVEC control group (P<0.05).At 5 days, HUVEC experimental group, as compared with the HUVEC control, showed a significantly increased number of newly formed vascular branches (7.36 vs 4.25/superbead, P<0.01) and relative length of the blood vessels (3.12 vs 1, P<0.01).Conclusion Linc00467 promotes tumor cell proliferation and angiogenesis and is highly expressed in the lung adenocarcinoma tissue, which is correlated with the tumor size and vascular invasion and suggests that Linc00467 could be a potential biomarker and therapeutic target.