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Objective To establish a Real-time quantitative PCR method (qPCR) for the detection of diatom UPA barcoding genes and evaluate its application in the drowning diagnosis. Methods The homologous sequences of diatoms UPA gene was obtained by Blast from GeneBank, based on which the universal primers for diatoms were designed. DNA were extracted from 2 common human symbiotic bacteria (Escherichia coli, Bifidobacterium longum), 3 species of planktonic bacteria, 15 species of planktonic algae, tissue samples (lung, liver and kidney) from human cadavers (28 drowning victims, 1 victims by non-drowning in the water, 3 victims deaths on land) in 32 cases. The specificity, sensitivity and repeatability of the designed primers were tested. The positive rates of diatoms detection in the drowning cases were calculated. The results of the real-time quantitative method were evaluated comparatively by Microwave Digestion-Vacuum Filtration-Automated Scanning Electron Microscopy (MD-VF-Auto SEM) and PCR-Capillary Electrophoresis (PCR-CE). Results The results showed that the primers UPA99 had strong specificity for the diatomaceae (Synedra radians, Navicula sp., Melosira varians, Cyclotella sp. and Nitzschia sp.) DNA. The melting curve of the amplified product was smooth; the peak was narrow; the melting temperature was (87±1)℃. The sensitivity of qPCR method was 1.56×10-5ng/μL with the detection range of 1.56×102ng/mL~1.56×10-5ng/μL, in contrast with the PCR-CE method (1.56×10-3ng/μL). This real-time PCR method showed high repeatability and stability with the coefficient of variation less than 2%. The detection rate of lung, liver and kidney was 89.3%, 71.4% and 64.3% respectively. Conclusion The established qPCR method, based on the universal primers designed for diatom UPA gene, has high specificity, high sensitivity and good repeatability. With a promising prospect for application, qPCR is suitable for drowning diagnosis.
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A method was developed for the rapud screenung and confurmatuon of 18 perfluorunated compounds ( PFCs) and 21 precursors un fush muscle by solud phase extractuon and luquud chromatography coupled wuth quadrupole/exactuve orbutrap mass spectrometry (LC-Q Exactuve Orbutrap MS). In thus method, the sample was furstly extracted wuth acetonutrule-water (90∶10, V/V), cleaned-up by solud-phase extractuon usung Oasus PRuME HLB column. Then, 39 target compounds were separated on a BEH C18(2. 1 mm×100 mm, 1. 7 μm) column, and the background unterference caused by LC system was elumunated by a short C18 HPLC column located between the muxer and the auto sampler. In the process of quantufucatuon and qualufucatuon, a full MS/dd-MS2 experument was adopted un mass spectrometry acquusutuon, accompanued wuth the full scan MS date for quantufucatuon, as well as the date dependent MS2 product uon spectra for qualufucatuon. The mass accuracy error was less than 3×10-6, and the calubratuon curves were lunear well wuth correlatuon coeffucuent over 0. 99. The lumuts of detectuon ranged from 0. 02 to 0. 50 μg/kg. The average spuked recoverues for 39 target compounds were between 61 . 7% and 122 . 0%, wuth relatuve standard deruvatuons ( RSDs ) from 6 . 9% to 18 . 8%. The developed method was successfully applued to the sumultaneous determunatuon of 18 PFCs and 21 precursors un fush muscle wuth satusfactory results.
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Objective To investigate the effect of coenzyme complex on the cardiac and renal functions of patients with type 2 cardiorenal syndrome. Methods Sixty-two patients with type 2 cardiorenal syndrome were enrolled in from June 2013 to December 2014 in Zhujiang Hospital , Southern Medical University. These patients divided were into routine group (n = 31) and coenzyme complex (n = 31). The therapy scheme of coenzyme complex group was on the basis of routine group with coenzyme complex intravenous drip , 400 U/day for 2 weeks. The cardiac function was determined by New York Heart Function Assessment, and the cardiac ultrasound, the levels of BNP. The renal function was determined by serum creatinine and urine volume. Results Compared with routine group, the rates of NYHA Ⅲ and Ⅳ were reduced, the LVEF levels were increased and the levels of BNP were increased (P < 0.05). The serum creatinine was decreased and urine volume were increased in the coenzyme complex group (P<0.05). Conclusion Coenzyme complex could improve the cardiac and renal functions of the patients with type 2 cardiorenal syndrome.
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<p><b>BACKGROUND</b>The insulin-like growth factor signaling pathway plays an important role in the modulation of cell growth and proliferation. The aim of this study was to investigate the role of polymorphisms of the insulin-like growth factor 2 (IGF2) and IGF-binding protein 3 (IGFBP3) genes, which encode key proteins of this pathway, as risk factors for gastric carcinoma (GC).</p><p><b>METHODS</b>A case-control study including 404 histologically confirmed GC patients and 424 healthy controls of the same ethnicity was conducted to retrospectively investigate the genetic polymorphisms of two genes, IGF2+820A>G (rs680) and IGFBP3 A-202C (rs2854744). Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using Logistic regression.</p><p><b>RESULTS</b>The IGF2 genetic variants examined contributed to GC risk individually (OR, 1.26; 95% CI, 1.08-1.46). The genotype frequencies of IGFBP3 A-202C were not significantly different between the cancer cases and controls (P > 0.05). Compared to the IGF2 AA genotype, carriers of one variant combined genotype were more pronounced among young subjects (<60 years), male subjects, never smokers, and those with a family history of cancer (OR = 1.36, 95% CI = 1.09-1.72, P < 0.05; OR = 1.61, 95% CI = 1.28-2.08, P < 0.05; OR = 1.46, 95% CI = 1.11-1.98, P < 0.05; OR = 1.53, 95% CI = 0.91-2.6, P < 0.05; respectively). Moreover, when the combined effects of the risk genotypes were investigated, significant associations were detected between highrisk genotypes in IGF2 and IGFBP3 (OR, 2.47; 95% CI, 1.75-3.49).</p><p><b>CONCLUSIONS</b>Our results suggest that polymorphic variants of the IGF2 genes modulate gastric carcinogenesis. Moreover, when the IGF2 and IGFBP3 variants are evaluated together, a greater effect on GC risk is observed.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Case-Control Studies , China , Genetic Predisposition to Disease , Genetics , Genotype , Insulin-Like Growth Factor Binding Protein 3 , Genetics , Insulin-Like Growth Factor II , Genetics , Logistic Models , Polymorphism, Genetic , Genetics , Stomach Neoplasms , GeneticsABSTRACT
Aim To explore the inhibitory effects and immunoregulatory activity of the extract from Eupolyphaga sinensis Walker (EFP) on the growth of liver cancer H22 in mice.Methods The chemical composition of extract from Eupolyphaga sinensis Walker(EFP)was separated with water extraction methods.The tumor bearing mouse model was constructed by injecting tumor cells subcutaneously.The EFP was given to the mouse by oral and hypodermic injection to observe its antitumor activity and immunoregulatory activity in vivo.The antibody level of anti-H22 in serum was measured with ELISA methods.The weight,tumor weight,spleen index,liver index were noted,and series of enzymes,such as,MDA,SOD,GPT and GOT activity of mouse serum and liver were measured.Results The extracts from Eupolyphaga sinensis Walker markedly inhibited the proliferation of tumor H22.The EFP constituent purified might be the main antitumor active part of Eupolyphaga sinensis Walker as shown by antitumor test in vitro.After treatment with EFP,the general condition of the test mice was much better than that of control mice,with spleen index,liver index increasing more obviously,antibody level of anti-H22,and MDA,SOD,GPT and GOT activity increasing more,and tumor growing more slowly.Conclusion EFP may be the main antitumor active part of Eupolyphaga sinensis Walker. EFP also has antitumor effect in vivo.EFP can exert antitumor effect in vivo by enhancing antibody level of anti-H22 and the series enzymes to be related to immunity activity.