1.Development and Application of Metabonomics in Forensic Toxicology
Journal of Forensic Medicine 2015;(3):219-226
Metabonom ics is an im portant branch of system biology follow ing the developm ent of ge-nom ics, transcriptom ics and proteom ics. It can perform high-throughput detection and data processing with multiple param eters, potentially enabling the identification and quantification of all sm all m etabolites in a biological system . It can be used to provide com prehensive inform ation on the toxicity effects, toxi-cological mechanisms and biom arkers, sensitively finding the unusual m etabolic changes caused by poi-son. This article mainly review s application of m etabonom ics in toxicological studies of abused drugs, pesticides, poisonous plants and poisonous anim als, and also illustrates the new direction of forensic toxi-cology research.
2.Development and Application of Metabonomics in Forensic Toxicology.
Journal of Forensic Medicine 2015;31(3):219-226
Metabonomics is an important branch of system biology following the development of genomics, transcriptomics and proteomics. It can perform high-throughput detection and data processing with multiple parameters, potentially enabling the identification and quantification of all small metabolites in a biological system. It can be used to provide comprehensive information on the toxicity effects, toxicological mechanisms and biomarkers, sensitively finding the unusual metabolic changes caused by poison. This article mainly reviews application of metabonomics in toxicological studies of abused drugs, pesticides, poisonous plants and poisonous animals, and also illustrates the new direction of forensic toxicology research.
Animals
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Biomarkers
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Forensic Toxicology/methods*
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Humans
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Metabolomics/methods*
3.Relationship between temperature of cornea and evaporative dry eye
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(02):-
Objective To investigate the impact factors for temperature of cornea(TOC) and analyse the relationship between TOC and evaporative dry eye. Methods Two hundred and fifty-seven patients(405 eyes) with normal lacrimal secretion received dry eye tests.Patients were divided into positive group and negative group according to the results,and were randomly subdivided into 4 groups with different environment temperature(T) and relative humidity(RH).For all eyes,TOC,body surface temperature(TBS) of forehead and center corneal thickness(CCT) were measured right after blinking.The impact factors for TOC and the differences in TOC between positive group and negative group were analysed.Results TOC was positively correlated with TBS(r=0.89),T(r=0.75) and RH(r=0.60)(P
4.Clinical Observation on General Anxiety Disorder Treated by Acupuncture plus Herbal Medicine
Journal of Acupuncture and Tuina Science 2007;5(4):234-237
To investigate the efficacy of the formula composed of the herbal drugs to soothe the liver, regulate qi, clarify the heart and eliminate vexation plus acupuncture for general anxiety disorder. Methods: Forty cases in the treatment group were treated with acupuncture plus herbal formula and 38 cases in the control group were treated with oral Doxepin. The assessment was conducted by Self-rating Anxiety Scale (SAS) and the Treatment Emergent Symptom Scale (TESS) before, during and after treatment. Results: The total effective rate was respectively 82.50% in the treatment group and 84.21% in the control group, with a significant difference in SAS assessment (P<0.01) between the two groups before and after the treatment.There were no significant differences in the curative and remarkable effective rate, total effective rate and SAS assessment between the two groups (P>0.05), but the side effect was higher in the control group than in the treatment group (P<0.01). Conclusion: Acupuncture plus herbal formula can have a precise effective effect for extensive anxiety neurosis with mild toxic side effects.
5.THE PRELIMINARY ESTABLISHMENT OF DEGRADATIVE REACTION SYSTEMS FOR DYES BY TRAMETES HIRSUTA
Hui-Rong LI ; Lin ZHANG ; Yan SHEN ;
Microbiology 1992;0(05):-
The growth-propagation and decolorization-degradation systems for the model dy e Biebrich Scarlet by Trametes hirsuta were established preliminar ily. It was showed that 30℃ and the static culture were better; the effects of compo nents in culture media on the efficiencies of decolorization and degradation wer e not significant; considering the convenience of observing the changes of dye c o lors and shortenig the culture period, the potato liquid medium had the advantag e of others as a preferable medium for reaction systems of Trametes hirsuta . The decolorization and degradation of all dyes Biebrich Scarlet and Direct Deep Blu e L-3RB, Reactive Blue X-BR, Basic Violet 5BN, and Methylene Blue by Tramete s hirsuta were better.
6.Evaluation of Brodifacoum-induced Toxicity by Metabonomics Approach Based on HPLC-TOF-MS
Hui YAN ; Xianyi ZHUO ; Baohua SHEN ; Ping XIANG ; Min SHEN
Journal of Forensic Medicine 2017;33(3):247-251,257
Objective T o analyse the m etabolic changes in urine of rats w ith brodifacoum intoxication, and to reveal the m olecular m echanism of brodifacoum-induced toxicity on rats. Methods B y establish-ing a brodifacoum poisoning rats m odel, the urine m etabolic profiling data of rats w ere acquired using high performance liquid chromatography-timeofflightmassspectrometry (HPLC-TOF-M S).The orthogo-nal partial least squares analysis-discrim ination analysis (O PLS-D A ) w as applied for the m ultivariate statistics and the discovery of differential m etabolites closely related to toxicity of brodifacoum . Results O PLS-D A score plot show ed that the urinary m etabolic at different tim e points before and after drug adm inistration had good sim ilarity w ithin tim e period and presented clustering phenom enon. C om paring the urine sam ples of rats before drug adm inistration w ith w hich after drug adm inistration, tw enty-tw o m etabolites related to brodifacoum-induced toxicity w ere selected. Conclusion T he toxic effect of brodi-facoum w orked by disturbing the m etabolic pathw ays in rats such as tricarboxylic cycle, glycolysis, sphin-golipid m etabolism and tryptophan m etabolism , and the toxicity of brodifacoum is characterized of accu-m ulation effect. The m etabonom ic m ethod based on urine H PLC-TO F-M S can provide a novel insight into the study on m olecular m echanism of brodifacoum-induced toxicity.
7.Effects of transfected gut-enriched Krüppel-like factor on growth of human gastric carcinoma cell line SGC-7901 and its xenograft in nude mice
Hui YAN ; Weichang CHEN ; Jiannong CEN ; Hongjie SHEN ; Xiaofei QI
Chinese Journal of Digestion 2011;31(1):30-35
Objective To investigate the antitumour effects of transfected gut-enriched Krüppellike factor(GKLF) on human gastric carcinoma cell line SGC-7901 in vitro and in vivo. Methods The expression of GKLF mRNA and protein in human gastric carcinoma cell line SGC-7901 were detected before and after transfection by real-time fluorescence quantitative PCR and Western blot,respectively. Proliferation and invasion in SGC-7901 were measured respectively by MTT assay, flow cytometry, colony formation assay and cell invasion assay after transfected with GKLF. The growth of xenograft was observed, the microvessel density(MVD) of xenograft tissue was determined by immunohistochemistry. Results The GKLF mRNA and protein in SGC-7901 were overexpressed after transfected with GKLF(P<0.05). The proliferative speed of SGC7901-pcDNA3.1-GKLF group was markedly lower than that of SGC-7901 and SGC7901-pcDNA3.1 groups (P<0.05). Transfected with GKLF caused part of the G0/G1 arrest, decreased clone formation rate and the invasion ability (P<0.05). The growth speed of xenograft in SGC7901-pcDNA3.1-GKLF group was lower, the weight and MVD of xenograft tissue in SGC7901-pcDNA3. 1-GKLF group were less (P< 0. 05).Conclusion Transfected with GKLF maysuppress proliferation and invasion in human gastric carcinoma cell line SGC-7901, inhibit the growth and the angiogenesis of xenograft in nude mice.
8.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*
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Humans
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Limit of Detection
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Poisoning
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Reproducibility of Results
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Tandem Mass Spectrometry/methods*
9.Determination of cocaine and benzoylecgonine in guinea pig's hair after a single dose administration by LC-MS/MS
Qiran SUN ; Ping XIANG ; Hui YAN ; Min SHEN
Acta Pharmaceutica Sinica 2008;43(12):1217-1223
A sensitive LC-MS/MS method to determine cocaine and its major metabolite benzoylecgonine in guinea pig's hair has been established. About 20 mg of decontaminated hair sample was hydrolyzed with 0.1 mol·L-1 HCl at 50 ℃ overnight, in the presence of cocaine-d3 and benzoylecgonine-d8 used as internal standards, and then extracted with dichlormethane. The analysis was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Positive electrospray ionization (ESI+) and multiple reactions monitoring (MRM) mode were used. The limit of detection (LOD) for cocaine and benzoylecgonine was 1 pg·mg-1. The calibration curves of extracted standards were linear over the range from 5 pg·mg-1 to 250 pg·mg-1 (r2≥0.999 7). The method was validated and applied to the analysis of guinea pig's hair after a single dose administration of cocaine hydrochloride. Cocaine and benzoylecgonine were not only detected, but also quantified in guinea pigs hair.
10.Hypoxic preconditioning inhibits apoptosis of bone marrow mesenchymal stem cells through overexpressing Pim-1
You ZHANG ; Weiya YAN ; Zhengya SHEN ; Junjie YANG ; Jie HUI
Chinese Journal of Tissue Engineering Research 2016;20(14):1989-1998
BACKGROUND:Bone marrow mesenchymal stem cel s have a low survival rate after implanted into the ischemic myocardium. However, hypoxia preconditioning (HPC) may enhance bone marrow mesenchymal stem cel proliferation and promote its survival rate. OBJECTIVE:To explore whether Pim-1 is involved in HPC protecting against apoptosis of bone marrow mesenchymal stem cel s and the relevant mechanism. METHODS:Bone marrow mesenchymal stem cel s were respectively subjected to HPC for 0, 6, 12, and 24 hours. The expression of Pim-1 and apoptosis-related genes were detected by RT-qPCR and western blot. Then, the best hypoxic preconditioning time was determined as 12 hours. Then, bone marrow mesenchymal stem cel s were assigned to one of the fol owing groups:control (without HPC), 12-hour HPC, 12-hour HPC+Pim-1 inhibitor groups. Flow cytometry analysis was used to detect the cel apoptosis, Transwel assay to analyze the cel migration ability in each group, and JC-1 kit to detect mitochondrial membrane potential. Animal models of myocardial infarction were established. One week after modeling, bone marrow mesenchymal stem cel s were given via multi-point injection around the infarct zone of rats. Two weeks after modeling, heart tissues of rats were taken and sliced fol owed by DiI staining to calculate the survival rate of bone marrow mesenchymal stem cel s. Additional y, rat cardiac function was assessed by echocardiography prior to and after modeling as wel as at 4 weeks after cel transplantation. RESULTS AND CONCLUSION:At 12 hours after HPC, the expression of Pim-1, p-Akt and Bcl-2 gene in the infarct region was significantly increased, but the expression of caspase-3 and Bax was significantly decreased. Compared with the control group, cel viability in the 12-hour HPC group was increased very significantly at 1 week after cel transplantation (P<0.001), the migration and anti-apoptosis ability were enhanced significantly (P<0.01) and the cardiac function of rats was significantly improved in the 12-hour HPC group (P<0.05). Al of these protective effects were blocked by the Pim-1 inhibitor. These findings indicate that HPC can protect bone marrow mesenchymal stem cel s from apoptosis through activating Akt and up-regulating Pim-1, and thereby improve the therapeutic effect of bone marrow mesenchymal stem cel transplantation on ischemic heart diseases.