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An ultra-high performance liquid chromatography-tandem mass spectrometry ( UHPLC-MS/MS ) method was developed and validated for the simultaneous determination of 9 kinds of trace endocrine disrupting chemicals in biological samples using ultrasonic-assisted extraction followed by purification with gel permeation chromatography ( GPC) and silica gel columns. The sample extracts were purified by Bio beads S-X3 GPC columns with cyclohexane/ethyl acetate (1:1, V/V) as mobile phase, and the target compounds were eluted in the fraction of 12-28 mL retention volume. Electrospray ionization source operated in positive mode and atmospheric pressure chemical ionization source operated in negative mode were used for mass spectrometric detection. Data acquisition was carried out in multiple reaction monitoring mode. Recoveries were predominately within 65 . 2%-118 . 0%. Method quantification limits were 0 . 1-9 . 7 ng/g dw ( dry weight ) . This method was successfully applied to the analysis of the target endocrine disrupting chemicals in carps collected from the Pearl River. with the exception of carbanilide and triclocarban, the rest analytes were detected in fish tissue samples, with the concentrations varied within the range of 0. 1-22. 6 ng/g dw.
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Objective To observe changes of cell cycle after stable silencing of SNCG gene in human endometrial carcinoma HEC-1A cells.Methods Flow cytometry was used to observe the changes of SNCG stable silencing HEC-1A cell growth cycle.The percentage of G2/M phase of HCE-1A cells after acridine orange staining was observed by laser scanning confocal microscopy.Results Flow cytometry results showed that the group of SNCG stable silencing in G1 and G2/M phase of the cells increased,but the percentage of cells in the S phase were decreased,the difference were both statistically significant (P ＜ 0.05).The results of cells after confocal microscopy observation of acridine orange staining,the group of SNCG stable silencing were observed in G2/M phase high percentage were statistically significant (P ＜ 0.05).Conclusion Two experimental methods commonly validated that the cell cycle of SNCG stable silencing on HEC-1A cells was arrested in G2/M phase,suggesting that SNCG gene is closely related to the cell growth cycle of endometrial carcinoma.
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Objective To investigate the relationship between renal vascular resistance index( RI) and serum Apelin level in type 2 diabetic mellitus ( T2DM )?Methods Seventeen cases newly diagnosed T2DM patients with RI increased but without microalbuminuraia in Minhang Hospital Affiliated to Fudan University from December 2011 to December 2014 were selected as observation group,17 newly diagnosed T2DM patients with RI normal during the same period were selected as control group?Fasting plasma glucose(FPG),glycosylated hemoglobin A1C(HbAlC),blood lipids and fasting insulin(FINS),hepatic functional and renal function were tested in all the subjects?Serum Apelin level was detected by enzyme?linked immunosorbent assay?Results Compared with control group,serum Apelin level was significantly higher in observation group((179?2±122?4)μg/L vs?(56?7±50?6) μg/L,t=3?814,P<0?05)?Partial correlation analysis showed that the RI was positive correlated with Apelin ( r= 0?364, P= 0?040 )?Conclusion The serum Apelin levels elevated in newly diagnosed T2DM patients with RI increased, and RI is positively correlated with Apelin, which indicate that Apelin play an important role in the pathophysiology of early renal damage in patients with TsDM.
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This paper was to explore the relationship between plasma omentin and obesity and insulin resistance in patients of polycystic ovary syndrome(PCOS).The results showed that there were statistically significant differences in serum omentin levels among control group,PCOS-normal weight group,and PCOS-overweight group [(115.29 ± 33.92),(111.12 ± 34.50),and (42.85 ± 19.39) pg/ml,P＜ 0.05].Omentin were negatively correlated with homeostasis model assessment for insulin resistance(HOMA-IR) and body mass index(BMI).In the multiple stepwise regression analysis,only omentin was negatively correlated with BMI.Omentin may be involved in the development of PCOS by affecting obesity.
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Objective To study the occurrence of BRCA1 MSI in endometrial cancer and its relationship with clinic pathologic features; to explore the correlation between MSI and protein expression in BRCA1 gene. Methods Application of PCR-SSCP and DNA sequence analysis method was used to study D17S579 and D17S1349 in 49 sporadic endometrial cancer tissues, 20 cases with endometrial atypical hyperplasia and 28 cases with normal endometrial tissues. Results In the total samples of D17S579 and D17S1349, the three groups were significantly different: 34.69%(17/49) in the endometrial cancer group, 10%(2/20) in the endometrial atypical hyperplasia group and 7.14%(2/28) in the normal endometrium group (χ2= 11.208, P = 0.004). BRCA1 MSI positive rate related to the pathology grade and clinical stage, but no relationship was found in muscular infiltration depth, lymph node metastasis and histopathology types. In the endometrial cancer group, BRCA1 MSI positive rate and BRCA1 protein expression were in moderate correlated negatively (r = -0.779, P = 0.000). Conclusion BRCA1 MSI might play a role in the development of endometrial cancer, and low expression of BRCA1 protein. BRCA1 MSI might be associated with pathology grade and clinical stages in EC.
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Objective To investigate the situation of subjective well-being feeling of type 2 diabetes patients and its influential factors.Methods One hundred and twenty-seven patients with type 2 diabetes were recruited as our subjects and all of them were performed the questionnaire investigation about well-being feeling.Influential factors were analyzed through several statistic methods.Results The index of subject wellbeing ranged from 3.70 to 13.70 and the average was 10.67 ± 2.10 of all type 2 diabetes patients.Of which The index of total affectation was 5.11 ± 0.92 and the range was from 2.63 to 6.63.The score of life satisfaction was 5.58 ± 1.22,and the range was from 1.10 to 7.70.Index of subjective well-being significantly associated with education,family income,therapy approach,HbA1 c level,complication,medicine burden.However there was no correlation with gender,age,diabetes duration.Conclusion The main types of subjective well-being feeling of patients with type 2 diabetes were positive effect and positive experience.Many factors could be the influence factor to well-being.Measures should be taken to improve Subjective well-being of type 2 diabetes patients.
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In order to study the functions of the HSPs (Heat shock proteins) of Eimeria tenella, we cloned a novel gene (which designated EtHSP) coding HSP of Eimeria tenella by RT-PCR and RACE (Rapid-amplification of cDNA ends). The full-length cDNA sequence of EtHSP was 1802 bp, containing a 1455 bp ORF (Open reading frame) (GenBank Accession No. FJ911605) encoding a deduced protein of 484 amino acids. Real-time PCR revealed that the mRNA level of EtHSP was much higher in sporozoites of E. tenella than other developmental stages (unsporulated oocysts, sporulated oocysts and merozoites). We constructed the recombinant plasmids pET28a(+)-EtHSP, then transformed it into E. coli BL21(DE3) for expression. SDS-PAGE indicated that the fusion protein was expressed in included bodies, with peak expression 6 h after induction by IPTG Western blotting revealed that the protein was specifically recognized by polyclonal antibodies against E. tenella, showing that the fusion protein was native antigen.
Subject(s)
Animals , Male , Rabbits , Chickens , Eimeria tenella , Genetics , Metabolism , Escherichia coli , Genetics , Metabolism , Heat-Shock Proteins , Genetics , Allergy and Immunology , Metabolism , Inclusion Bodies , Metabolism , Molecular Sequence Data , Open Reading Frames , Genetics , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Metabolism , Sequence Analysis, ProteinABSTRACT
Objective:To probe the expression of Fragile histidine triad(FHIT)and p21 protein in Vulvar Condyloma acuminatum(VCA) and vulvar intraepithelial neoplasia(VIN).Methods:Immunohistochemistry was used to detect FHIT and p21 expression in 51 cases of VCA,39 cases of VIN and 20 cases of normal vulvar epithelium.Results:The positive rate of FHIT in VCA and VINⅠ～Ⅱ、VINⅢwas obviously lower than which in normal tissue(P=0.000 1);there was no statistic significance in the positive rate of FHIT expression between VINⅢand VCA(P=0.592);the positive rate of p21 in normal vulva,VIN and VCA has statistic significance;there was no statistic significance in the positive rate of p21 expression between VIN and VCA(P=0.793).Conclusion:(1)FHIT and p21 expression are associated with the development and progression of VCA and VIN(.2)The abnormality of FHIT gene is a early event in the development and progression of vulvar cancer.