2-Amino-3-hydroxylpyridinE-H2O2-horseradish peroxidase (HRP) voltammetric enzymE-linked immunoassay based on N-heterocyclic substrate has been successfully applied for the detection of carcionembryonic antigen(CEA) in human serum. 2-Amino-3-hydroxylpyridine is oxidized with H2O2 catalyzed by HRP, and the resulting electroactive product produces a sensitive voltammetric peak at the potential of 0.36 V(vs. SCE) in Britton-Robinson (B-R) buffer solution. By this voltammetric peak, free HRP can be measured and immunoassay of HRP label can be developed. The enzymE-catalyzed reaction conditions and voltammetric detection conditions have been optimized, and the electrode procedure of the enzymatic product was investigated. The selected optimum reaction conditions were that the reaction medium was pH 6.0 B-R buffer solution for 10 mL reaction solution containing 1.0 mL of 0.2 mol/L B-R buffer solution, 3.0 mL of 8.0 mmol/L 2-amino-3-hydroxylpyridine solution and 1.5 mL of 0.5 mmol/L H2O2 solution, and the reaction time was 30 min at 37 ℃. The optimum detection conditions were that the supporting electrolyte was pH 7.0 B-R buffer solution for 10 mL of the overall detection solution containing 5 mL of reaction solution and 1.0 mL of 0.2 mol/L B-R buffer solution. The optimum instrumental conditions for the detection were chosen as follows: the initial potential, 0.00 V; the final potential, -0.80 V; the potential scanning rate, 400 mV/s; the mercury drop standing time, 7 s. Under the optimum conditions, the linear range for detection of free HRP was 4.0×10-4-1.0 μg/L with a detection limit of 0.12 μg/L. Based on the new immunoassay system, the linear range of the detection to CEA was 0.50-80 μg/L and the detection limit was 0.50 μg/L, which is 10 times lower than that of traditional spectrophotometric enzymE-linked immunosorbent assay method. The 2-Amino-3-hydroxylpyridinE-H2O2-HRP voltammetric enzymE-linked immunoassay new system exhibits excellent performance having wider linear range and lower detection limit. The new method is inexpensive and simple. It has a great potential in clinical diagnosis.

Objective To investigate the influence of wound treatment methods on residual ovarian reserve function after laparoscopic ovarian cystectomy.Methods A total of 120 patients with ovarian cyst who underwent laparoscopic ovarian cystec‐tomy in the hospital were randomly divided into three groups in terms of different wound treatment methods:electric coagulation group ,ultrasound group and suture group ,with 40 cases in each group.The levels of follicle stimulating hormone(FSH) ,luteini‐zing hormone(LH) ,and estradiol(E2 ) were detected in the blood before and immediately after the operation ,2 days after men‐struation at postoperative 3 and 6 months ,and the number of ovarian reserve follicles determined by transvaginal ultra‐sound.The changes of menstruation and clinical manifestations of the patients were recorded at the same time.Results The lev‐els of LH ,E2 and FSH were dramatically changed in electric coagulation group ,which was followed by the ultrasonic group.Patients in the suture group had the smallest changes of these indices and recovered quickly(P<0.05).In the electric co‐agulation group relative to the other two groups ,the number of ovarian reserve follicles was significantly decreased and the inci‐dence of adverse reactions was profoundly increased.Conclusion Electric coagulation greatly adversely affects the residual ovar‐ian reserve function after laparoscopic ovarian cystectomy ,and ultrasonic and suture hemostasis techniques can help to protect o‐varian function.

Objective To evaluate the effectiveness of a project for treating TB/HIV co-infection(phase 1,2007-2008)supported by Global Fund in Wolong district.Method Data on project implementation in 2007-2008 were collected and analyzed.Results A total of 916 TB patients were diagnosed in 2007-2008,of whom 847 cases were detected for HIV antibody,with a detection rate of 92.47%.A total of 368 people living with HIV/AIDS were identified by screening 1790 persons/times,with a screening rate of 94.36%.Among them 55 cases of active tuberculosis were co-infected with HIV,of whom 3 were smear positive.After aniti-TB chemotherapy marked symptomatic improvement was witnessed in 37 cases;the curative rate for new smear positive cases reached 66.7% and the remission rate was 70.0%.Conclusion Satisfactory results were achieved in preventing and treating TB/HIV co-infection in Wologn district,and the experience gained in implementing the phase 1 project will provide useful information for implementing the project for the next phases.

Objective ERP research has always been the hot spot of the n~ve electrophysiology research.A gTowing body of research suggests that ERP can effectively reflect the cognitive function in patients with depression,evaluate the risk of suicide.This paper reviews the ERP related components with depression and related research.On the basis of the application of ERP in the depression research discusses limitations and prospects on the later development.Methods The articles were retrievaled from databases,such as P UBMED,Foreign medical information resources retrieval platform,CNKI,Wanfang database,ect,from 2013-06 to 2015-01.Index strategy TI:TI:[depression OR affective disorder] AND [suicide OR suicidal] All field:event related potential.Reading and narrowing the scope of the articles,we should understand the experimental design,such as Sample company have suicidal ideation and (or) behavior of patients with depression.Potential induced way,ERP related component detection method,etc.The articles included 43 in English and 15 in Chinese.Read the full text and retain nearly five years about depression and related literature.Results Eventually,the article includes 34 articles and 3 of in Chinese.Conclusion Associated with suicidal ideation and (or) behavior of the existing cognitive dysfunction in patients with depression,ERP related components such as:abnormal P300 and CNV,MMN.N400 have not found abnormal.This article only from the Angle of depression and related research are reviewed in this paper the characteristics of ERP,ERP of other mental illnesses among related research,and to be perfect in the future.

Objective Recent years,epigenetics research mainly focused on DNA methylation.A growing body of research indicated that DNA methylation may play vital role in etiology of affective disorder.With brief introduction of DNA methylation molecular mechanisms and relevance of DNA methylation to affective disorder,this review focuses on some supporting evidence which are recently emerged in major depressive disorder (MDD) and bipolar disorder related gene DNA methylation.Methods We retrievaled related articles from databases,such as PUBMED,Foreign medical information resources retrieval platform,CNKI,Wanfang database,ect,from 2013-06 to 2015-01.Index strategy TI:[depression OR bipolar disorder OR affective disorder] AND All field:DNA methylation.Reading and narrowing the scope of the articles,the experimental design was understanded,such as DNA from either blood cell or brain parenchyma of sample,methods of methylation detection,gene detected and methylation position,etc.The articles included 168 in English and 32 in Chinese.The whole and retained the articles were red about MDD and bipolar disorder related gene DNA methylation in recent five years.Results Eventually,the article includes 29 articles and two of in Chinese.Most of them are receptor gene epigenetics research associated with disease.Conclusion MDD and bipolar disorder are associated with the higher of DNA methylation levels in multiple gene promoter region,such as GR,BDNF,5-HTR,SLC6A4 and the lower of COMT.This article only from the perspective of affective disorder reviewed the relationship between DNA methylation and diseases.None of other diseases epigenetics research,there is to be perfected in future.

Objective To investigate the relationship between peritoneal macrophages (PMAs) and inflammatory reaction in a rat model of severe acute pancreatitis (SAP). Methods Sprague-Dawley rats were randomly divided into control group and SAP group. To induce SAP in rats, 40 g/L sodium taurocholate (0.1 mL/100 g) was injected into the pancreatic duct through retrograde exposure of pancreatic bile duct in hepatic porta. One-third of rats were sacrificed at 3, 6 or 12 h after modeling. PMAs were extracted, and incubated for 24 h in a humidified 5% carbon dioxide incubator. The expressions of tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β) mRNA in PMAs were measured by semi-quantitative RT-PCR. The levels of TNF-α and IL-1β in culture medium and serum were evaluated.The histological changes of pancreas were examined. Rosults The expressions of TNF-α mRNA and IL-1β mRNA in PMAs were significantly higher in SAP group than in control group at each time point (P<0.01). The concentrations of TNF-α and IL-1β in culture medium and serum were significantly elevated in SAP group compared with control group (P<0.01). The histological analysis of pancreas indicated that the damage was more severe in SAP group than in cuntrol group (P<0.01). Conclusion PMAs secrete cytokines into pancreatitis-associated ascitic fluid, and this study demonstrates a correlation between SAP and the activation of PMAs.

ObjectivesTo explore the roles of mean corpuscular volume(MCV),mean corpuscular hemoglobin(MCH) and hemoglobin A2 (HbA2) in the laboratory screening of thalassemia,and to find optimal screening modality for different conditions.Methods From September 2008 to May 2011,1384 subjects underwent thalassemia screening at Department of Obstertrics andGynecology of Nanfang Hospital.Of them,1036 cases were diagnosed with thalassemia (408 α-thalassemia,608 β-thalassemia,and 20 αβ compound thalassemia,thalassemia group) and 348 without thalassemia,non-thalassemia group.All subjects were screened respectively for MCV,MCH and HbA2.Analyses were performed in all subjects to assess the sensitivity,specificity,positive predictive value,negative predictive value and diagnostic accuracy respectively associated with MCV,MCH and HbA2 alone,combination of MCV and MCH,and combination of MCV,MCH and HbA2.Results( 1 ) In the thalassemia group,the sensitivity of MCV alone was 92.9％ (379/408) for α thalassemia,99.3％ (604/608) for β thalassemia and 100.0％(20/20) for αβ compound thalassemia.In the non-thalassemia group,the specificity of MCV alone was 75.0％ (261/348).(2) In the thalassemia group,the sensitivity of MCH alone was 92.9％ (379/408) in α thalassemia,99.0％ (602/608) in β thalassemia and 100.0％ (20/20) in αβ compound thalassemia.In the non-thalassemia group,the specificity of MCH alone was 72.7 ％ (253/348).(3) The sensitivity of Hb A2 alone was 67.4％ (275/408) for α thalassemia,97.5％ (593/608) for 3 thalassemia,and 100％ (20/20) for α3 compound thalassemia while it's specificity was 72.4％ (252/348) in the non-thalassemia group.(4)With positive indexes of MCV,MCH and MCV + MCH,when HbA2 ＞ 3.5％ it had a high value in [β-thalassemia screening,but when HbA2 ＜ 2.5％ it had little value in α-thalassemia screening.(5) As a single marker,MCV and MCH had better sensitivity,specificity,positive predictive value,negative predictive value and diagnosis accuracy than HbA2.MCV + MCH was the best for overall screening,but for [β thalassemia screening,MCV + MCH + HbA2 was the best.ConclusionsMCV and MCH are suitable for epidemic screening in a large population,physical examination and premarital check-up.Hb electrophoresis andthalassemiagenediagnosisarerecommendedforsubjectswithpositiveMCVandMCH indexes.Diagnoses of α and β-thalassemia gene are recommended for pregnant women with positive MCV and MCH indexes.

Objective To construct an eukaryotic expression vector of human B-cell translocation gene 2 (BTG2), to express the FLAG-tagged BTG2 protein in HeLa cells,and to supply an experimental tool for investigating the function of BTG2 gene.Methods The full-length BTG2 fragment was obtained by PCR and inserted into the multiple cloning site of pcDNA3.1 (+)vector. Oligo DNA encoding FLAG tag was designed and inserted into pcDNA3.1(+)-BTG2 to construct another vector pcDNA3.1(+)-FLAG-BTG2.The HeLa cells were divided into pcDNA3.1(+)empty vector group,pcDNA3.1(+)-BTG2 group and pcDNA3.1(+)-FLAG-BTG2 group.The HeLa cells were transfected with recombinant plasmids.Western blotting using anti-FLAG antibody was performed to detect the expression of FLAG-BTG2 protein in HeLa cells.Results The sequence of the vector was verified by both BamH Ⅰ endonuclese digestion and DNA sequencing. The Western blotting analysis confirmed that FLAG-fused BTG2 was detected in pcDNA3.1(+)-FLAG-BTG2 group but not in empty vector or pcDNA3.1(+)-BTG2 groups. Conclusion The eukaryotic expression vector pcDNA3.1(+)-FLAG-BTG2 is successfully constructed and FLAG-tagged BTG2 protein is expressed in HeLa cells.

Employment related with the quality of life and the development of occupation of the people with mental retardation(MR).This paper compared the employment of MR native and abroad,suggested the strategy referencing to the supportive employment theory.

OBJECTIVETo detect potential mutation of F8 gene in a family affected with hemophilia type A.

METHODSInverse-shifting PCR (IS-PCR), next-generation sequencing (NGS), multiplex ligation-dependent probe amplification (MLPA), and short tandem repeat (STR) assays were used.

RESULTSIS-PCR showed that no inversion of F8 gene has occurred in the family. NGS detected no point mutation or small InDel in the proband, but suggested that the exon 2 of the F8 gene may be deleted. MLPA also showed that exon 2 of the F8 gene was absent in the proband, while the carriers were heterozygous for the deletion, though STR analysis yielded a paradoxical result.

CONCLUSIONNGS analysis has identified a large deletion of exon 2 of the F8 gene in a family affected with hemophilia A. Discretion is required when STR analysis was used for carrier screening and antenatal diagnosis. Combination of multiple methods can improve the accuracy for the detection of F8 gene mutations.

Child ; Exons ; genetics ; Factor VIII ; genetics ; Genetic Testing ; methods ; Hemophilia A ; Humans ; Male ; Pedigree ; Sequence Deletion ; genetics