Analysis for polymorphism of MTHFR C677T gene in Han population in Beijing / 临床检验杂志

Objective@#To explore the distribution characteristics of MTHFR gene polymorphism of Chinese Han population in Beijing, and analyze the differences of distribution of MTHFR gene polymorphism between Beijing area and the other parts of northern China. @*Methods@#MTHFR C677T gene was detected by PCR-gold magnetic particles chromatography. The distribution characteristics of MTHFR gene polymorphism of 3 945 healthy subjects from September 2014 to May 2018 detected in Peking Union Medical College Hospital were analyzed retrospectively. The distribution of MTHFR gene polymorphism in Chinese Han population in Beijing area was compared with the Han population of the other area from northern China by reviewing domestic and foreign literature databases. @*Results@#The frequencies of CC, CT and TT genotype of MTHFR C677T gene in the male population undergoing physical examination in Beijing were 23.3%, 50.5% and 26.2%, respectively, and the frequencies of C and T allele frequencies were 48.6% and 51.4%, respectively. The frequencies of CC, CT and TT genotype of MTHFR C677T gene in the female population were 22.7%, 49.4% and 27.9%, respectively, and the C and T allele frequencies were 47.4% and 52.6%, respectively. There was no difference in genotype frequency and allele freaaency between male and femal (P＞0.05). The frequencies of CC, CT and TT genotypes and allele frequencies of MTHFR C677T gene in the population of Beijing area were significantly different from those of Heilongjiang, Jilin, Hebei, Shandong and Henan provinces (P＜0.01). @*Conclusion@#There was no significant difference of polymorphism distribution of MTHFR C677T gene between male and female populations in Beijing. The distinct distribution characteristics of MTHFR gene in Beijing area should be presented.

Infection Modes and Subtypes of Human Papilloma Virus in Patients / 中国医学科学院学报

Objective To investigate the prevalence of human papilloma virus(HPV)subtypes in patients and to provide an evidence for the prevention and treatment of HPV infection and the development of HPV vaccine. Methods Multiplex PCR was used to detect HPV DNA in 6917 patients in Peking Union Medical College Hospital from January 1,2013 to June 30,2015.Totally 5586 patients entered the final analysis after the repeat samples were deleted.The total positive rate of HPV subtypes(including high-risk subtypes including HPV-16,18,31,33,35,39,45,51,52,56,58,59,and 68 and low-risk subtypes including HPV-6 and 11)and the infection status of different age were analyzed. Results The total positive rate of HPV was 36.29%(2027/5586).The positive rate of high-risk subtype was 24.92%(1392/5586)and low-risk subtype was 1.66%(93/5586).The positive rate of multiple was 9.70%(542/5586)and multiple high-risk subtype was 7.75%(433/5586).The positive rate of high-risk subtype and multiple were 25.52%(1366/5353)and 11.16%(26/233)in female and 9.99%(535/5353)and 3.00%(7/233)in male,there were significantly difference(χ=24.61,χ=12.45,all P<0.001).The positive rate of low-risk subtypes(3.86%,9/233)in males was significantly higher than that in females(1.57%,84/5353)(χ=5.84,P=0.007).The high-risk HPV subtype infection mainly was seen in patients aged 31-50 years and the low-risk HPV subtype infection mainly in patients aged 21-40 years.The age of multiple HPV infections from 31-40 years.The lowest turn negative rates of subtype were HPV52 and HPV58.The top three HPV subtypes with the highest positive rates were HPV52,HPV16,and HPV58.Conclusions The positive rates of HPV type are different between male and female patients.The males are mainly infected with low-risk subtypes,whereas the females with high-risk subtypes and the multiple HPV subtypes.The top three high-risk subtypes are HPV-52,16,and 58.HPV subtypes with the lowest secondary negative rates are HPV-52 and 58.HPV infection is mainly seen in young individuals.

Analysis on HLA B27 results detected by real-time fluorescence PCR method and flow cytometry / 国际检验医学杂志

Objective To compare the clinical application value of the real-time fluorescence PCR method and flow cytometry method for detecting human leukocyte antigen B27(HLA-B27).Methods Blood HLA-B27 level in 225 patients with suspected ankylosing spondylitis was detected by using real-time fluorescence PCR method and flow cytometry method.The detection results were compared and analyzed between the two methods.Results The results of 95.1% sample were identical detected by two methods without statistically significant difference(P>0.05).Taken the results of flow cytometry as reference, the sensitivity of real-time fluorescence PCR for detecting HLA-B27 was 94%, the specificity was 96%.Gene sequencing was performed if results of a sample detected by two methods were different, which was identical with the result detected by real-time fluorescence PCR.Conclusion Both methods for detecting HLA-B27 all have high sensitivity and specificity.Real-time fluorescence PCR method is more superior to the flow cytometry method in the results accuracy.

Establishment of MALDI TOF-MS Technique Platform for Detecting Cytochrome P4 5 0 Gene Polymorphism / 现代检验医学杂志

Objective To establish the MALDI-TOF-MS technique platform for detecting Cytochrome P450 gene polymor-phismfrom of patients.Methods Collected 53 EDTA anticoagulation peripheral blood samples from October 2013 to June 2014 in Peking Union Medical College Hospital.The whole genomic DNA was extracted from patients’peripheral blood. Used MALDI-TOF-MS to identify the SNP polymorphism of CYP2C9*2(rs1799853),CYP2C9*3(rs1057910),CYP2C19*2(rs4244285),CYP2C19*3(rs4986893),CYP2C19*4(rs28399504),CYP2C19*5(rs56337013)and CYP2C19*17 (rs12248560).To verify the above SNP polymorphism by Sanger sequencing method.Results The MALDI-TOF-MS could perform 53 samples on two cytochrome gene and 7 SNP locus simultaneously.In all the 53 patients,25 AG,6 AA and 22 GG genotypes were identified in CYP2C19*2(rs4244285),the allele frequency of A genotype was 34.9%.4 AG and 49 GG genotypes were identified in CYP2C19*3 (rs4986893),the allele frequency of A genotype was 3.8%.5 CA and 48 AA gen-otypes were identified in CYP2C9*3 (rs1057910),the allele frequency of C genotype was 4.7%.No mutation loci were i-dentified in CYP2C9*2 (rs1799853),CYP2C19*4 (rs28399504),CYP2C19*5 (rs56337013)and CYP2C19*17 (rs12248560).All the identification data were confirmed by Sanger sequencing.The coincidence rate was 100%.Conclusion The MALDI-TOF-MS technique platform for the cytochrome enzyme SNP was established.This platform has high throughput and accurate characteristics.It has important clinical application value for the treatment of personalized medicate.

Molecular diagnosis of thalassemia compound heterozygotes in PUMCH from 2012 to 2015 / 中华检验医学杂志

Objective To investigate the genotype distribution of thalassemia intermedia , major and compound thalassemia in Peking Union Medical College Hospital from 2012 to 2015. Methods Retrospectively 1 084 suspected thalassemia cases were analyzed in recent four years .Three common deletions of αglobin chain were detected by GAP-PCR.Three common point mutations of αglobin chain and seventeen common mutations of βglobin chain were identified by PCR reverse dot blot hybridization . Hemoglobin electrophoresis was carried out by Capillary Electrophoresis System .RBC associated parameters and morphology were analyzed by hematology analyzer and blood smear .Results 702 cases were confirmed to be thalassemia, and the positive rate was 64.76% (702 /1084).19 types of gene defects were detected. There were 4 types of gene defects in 23 case with α-thalassemia intermeida, including -α3.7 /--SEA , -α4.2 /--SEA , αCSα/--SEA and αQSα/--SEA , -α3.7 /--SEA to be the most common genotype (18 cases) .3 cases with β-thalassemia intermeida were confirmed and the genotypes were βCD 17(A→T) /β-29(A→G) , β-28(A→G) /β-28(A→G) andβIVS-Ⅱ-654(C→T) /βCD17(A→T) , respectively.There were also 1 βCD 41 -42(-TTCT) /βCD17(A→T) thalassemia major case. The genotypes of 2 HbE/β-thalassemia cases were βCD41 -42(-TTCT) /βE and βCD17(A→T) /βE.5 αβ-thalassemia including 2 βCD 41 -42(-TTCT) /βA compounded with αα/-α3.7 , 1βIVS-Ⅱ-654(C→T) /βA compounded with --SEA /αCSα, 1βCD17(A→T) /βA compounded with -α4.2 /ααand 1βCD 41 -42(-TTCT) /βA compounded with αCS α/αα.Rare and untypical haematological results were found , such as normal level HbA 2 and undetectable HbH, in compound heterozygosity with --SEA /αCS α and βIVS-Ⅱ-654(C→T) /βA. Conclusions The genotypes of thalassemia intermedia, major and compound thalassemia in Peking Union Medical College were highly variable .

Clinical Molecular Biology Analysis of Three Kinds of Sexually Transmitted Diseases in Patients / 现代检验医学杂志

Objective To analysis the related factors and the morbidity of patients with Chlamydia trachomatis(CT),Neisse-ria gonorrhea(NG),Ureaplasma urealyticum (UU)in Beijing Union Medical College Hospital.Methods Simultaneous Am-plification and Testing (SAT)was used to detect CT-RNA,NG-RNA,UU-RNA of patients from January of 2013 to Decem-ber of 2014.Results There were 6 262 cases in all and 2 412 cases in 2013,and 3 850 cases in 2014.The positive rate of CT-RNA,NG-RNA and UU-RNA were 7.29%,3.44% and 38.21% in 2013.The positive rate of CT-RNA,NG-RNA and UU-RNA were 8.17%,4.15% and 45.71% in 2014 respectively,which were higher than those of in 2013.There was a signifi-cant difference in UU-RNA positive rate between the two years (χ2 =12.66,P <0.01).The positive rate of CT-RNA,NG-RNA and UU-RNA were 8.15%,3.89% and 42.77% respectively in two years.The positive rate of UU-RNA in women’s (61.05%)was significantly higher than the men’s (24.50%)(χ2 =316.13,P <0.01).Most of the people with CT-RNA, NG-RNA and UU-RNA-positive were between 21 to 30 years old.The highest detection positive rate was in cervical swabs samples of UU-RNA.Conclusion There was a significantly increasing trend of the infection with UU.Male were more sus-ceptible to CT and NG.However,female were more susceptible to UU.There was the highest infection rate in youth (21~30 years)with CT,NG and UU.

Assessment of the consistency of five standardized cystatin C measurement systems / 中华检验医学杂志

Objective To assess the consistency of four standardized cystatin C particle-enhanced turbidimetric assay (PETIA) and one particle-enhanced nephelometric immunoassay (PENIA) measurement systems Methods Performance verification test was conducted according to CLSI EP 15-A2 and EP9-A2. Fourty serum samples in comparative test were obtained from the remaining serum samples of outpatients in Peking Union Medical College Hospital in March 2013.Fourty serum samples were tested on Olympus AU5400 automatic biochemical analyzer ( four PETIA Cys C reagents:Shanghai Jingyuan Co ., Ltd, Beijing Leadman Biochemistry Co ., Ltd, Beijing Strong Biotechnologies , Maker Biotechnology in Sichuan , and labelled as A, B, C, D respectively) and PENIA N Latex Cys C measurement system on Siemens BNⅡ(labelled as E).Correlation analysis were performed among four PETIA methods one PENIA method Differences of each detection system were compared in the medical decision level 1,2,3,4 mg/L.The reference material ERM-DA471/IFCC was measured by five systems and bias ( percentage bias ) was calculate for each system.Results Results of systems A, B, C, D, E were 1.29(0.89-2.43), 1.30 (0.96-2.59), 1.22(0.90-2.44), 1.27(0.96-2.47), 1.14(0.82-2.05)mg/L.Chart shows bias among these five systems was small when Cys C concentration was less than 4mg/L.PETIA method A, B, C, D correlated with their mean value well , with the average deviation from their mean value ( percent deviation) at -0.017 -0.031 mg/L ( -3.1%-2.1%), and all were less than allowed bias from the biological variation (3.4%).The deviation of PETIA method A, B, C, D with their mean value in medical decision level at -0.176 -0.178 mg/L.Systems A, B, C, D correlated well with the result of PENIA method system E , and the mean deviation ( percent deviation ) was at 0.278 -0.326 mg/L ( 12.6%-18.5%) , and the deviation ( percent deviation ) in the medical decision level 0.055 -1.079 mg/L (5.51%-26.98%).Bias of PETIA method A, B, C, D Cys C system measuringERM -DA471/IFCC ranged from 0.22 to 0.39 mg/L ( 3.9%-7.0%) , which exceeded the allowable range of the reference material target value, and were larger than the allowable bias from biological variation (3.4%).Bias ( percent ) of PENIA method system E was -0.1 mg/L ( -1.7%) , within the allowable range of ERM-DA471/IFCC target value .Conclusions The consistency of four assesed PETIA Cys C reagents was relatively ideal, and improved markably after being traced to ERM-DA471/IFCC.Besides, the results of PETIA were higher than those of PENIA .Bias among these five systems was small when Cys C concentration was less than 4 mg/L, and the bias became larger in higher Cys C concentration.

Effects of Microiontophoretically Applied Some Different Transmitter Receptor Antagonists on the Evoked Response of “Pain” Units of Hypothalamic Paraventricular Nucleus / 第二军医大学学报

Effects of microiontophoretically applied transmitter receptor antagonists on the evoked response of “pain” units of PVN in rats by sciatic stimulation were observed. The results showed: (l)The evoked response of 7 out of 38 PVN “pain” units could be blocked by atropine (7/38); 5 out of 27 by hexamethonium; 6 out of 31 by phentolamine (6/31); and 4 out of 25 by propranolol (4/25). Seventeen out of 52 were blocked by cyproheptadine (17/52), but 3 out of 52 were augmented by cyproheptadine (3/52). (2) The evoked response of the same “pain” unit could be blocked by two antagonists: the evoked response of 2 out of 27 PVN “pain” units could be blocked by atropine and cyprohepadine; 3 out of 20 by hexamethonium and cyproheptadine; 2 out of 25 by prepamolol and cyproheptadine. These results suggest that the noxious somatic input to PVN involves 5-HT, cholinergic and adrenergic transmitter receptor mechanisms and that the convergence of various transmitter systems on PVN “pain” unit is indicated.