1.A Chemical Inhibition Method in the Measurement of Mitochondria Isoenzyme of GOT
Chinese Journal of Clinical Laboratory Science 1985;0(04):-
The competitive inhibition of s-GOT was made by the amber-acid(the final concen-tration was 4.6?10~(?) mol/l).m-GOT was measured by Reitman's method.The recoveryrate was 102%.The range of 10~80 KU of m-GOT was linear.The mean value of m-GOTfor 155 healthy adults was 2.8?2.7((?)+SD),76.8% of them≤5 KU.There was no diffe-rence in sex.The accuracy,normal value were similar to those in the column chromatog-raphy and immuno-assay method.The result of 66 patients revealed that m-GOT could beused as the parameter of the degree of hepatic cellular damage in virus hepatitis and inmyocardial destruction.
2.Screening and identification of prognosis-related miRNA in hepatocellular carcinoma by the TCGA database
Dan SHI ; Bin XU ; ting Jing JIANG
Chinese Journal of Clinical Laboratory Science 2017;35(11):818-821
Objective MicroRNAs(miRNAs) play a critical role in the prognosis of liver cancer,especially in hepatocellular carcinoma (HCC).The aim of this study was to find the new prognosis-related miRNAs in HCC through data mining of The Cancer Genome Atlas(TC-GA).Methods The miRNA expression data with clinicopathological characteristics and prognosis of liver cancer patients were obtained from the TCGA data portal and then data analysis,integration and standardization were performed by R package.Moreover,univariate Cox model and high-dimensional analytical model BhGLM(Bayesian hierarchical generalized linear model) were used to analyze the miRNA expression profiles data.Results A total of 63 and 77 prognosis-related miRNAs were screened using Cox and BhGLM model respectively,and 18 miRNAs were screened by co-analysis of Cox and BhGLM.Among them 4 prognosis-related miRNAs such as miR-188,miR-576,miR-887 and miR-91 have never been reported in HCC.Conclusion On the basis of these data,co-analysis of Cox and BhGLM could be an effective method which screen and validate miRNAs as the new clinical prognostic and diagnostic biomarkers in HCC.
3.Expression and roles of long non-coding RNA ZFAS1 in non-small cell lung cancer
Xu ZHANG ; yan Xue ZANG ; mei Jian GU ; Peng ZHANG ; Wei LIANG ; yan Jing CHEN ; Lei PAN ; Hui QIAN ; rong Wen XU
Chinese Journal of Clinical Laboratory Science 2017;35(11):813-817,848
Objective To determine the expression levels of zinc finger antisense 1 (ZFAS1) in tumor tissues of non-small cell lung cancer (NSCLC) and investigate the biological roles of ZFAS1 in NSCLC.Methods The relative expression levels of ZFAS1 in tumor tissues of NSCLC patients were determined by using real-time fluorescent qRT-PCR.RNA interference was used to knock down ZFAS1 expression in A549 cells.The proliferations of A549 cells with ZFAS1 knockdown were determined by cell counting and cell colony formation assays.Flow cytometric analysis was used to determine the cellular cycle distribution and apoptosis of the A549 cells with ZFAS1 knockdown.Transwell migration and matrigel invasion assays were used to determine the migration and invasion of the A549 cells with ZFAS1 knockdown.The gene expression levels of cyclin D1,Bcl2,N-cadherin,ZEB1,Slug and Twist were determined by real-time fluorescent qRT-PCR.Results The mean expression levels of ZFAS1 in tumor tissues of NSCLC patients [0.01 (0.002 to 0.054)] were significantly higher than those in the adjacent non-cancerous tissues [0.002 (0.001 to 0.012)] (Z =-2.638;P < 0.01).After ZFAS1 knockdown,the proliferation of A549 cells was remarkably retarded (P < 0.01).The percentage of cells at G1 phase was increased while the cells at S phase was decreased (P < 0.01).The rate of apoptotic cells was significantly increased in A549 cells with ZFAS1 knockdown (P < 0.01).A549 cells showed decreased migration and invasion abilities after ZFAS1 knockdown (P <0.01).The expression levels of cyclin D1,Bcl2,N-cadherin,ZEB1,Slug and Twist genes were decreased in ZFAS1 knockdown A549 cells (P < 0.05).Conclusion ZFASI was highly expressed in the tumor tissues of NSCLC patients.ZFAS1 knockdown induced cell cycle arrest,cell apoptosis and suppression of epithelial-mesenchymal transition (EMT),leading to the inhibition of proliferation,migration,and invasion of NSCLC cells.
4.Prognostic value of SCC-Ag for cervical cancer
Yao XIAO ; jie Ying SHAO ; hua Jian JIN ; ting Jing JIANG ; ping Chang WU
Chinese Journal of Clinical Laboratory Science 2017;35(11):808-812
Objective To evaluate the prognostic significance of squamous cell carcinoma-associated antigen (SCC-Ag) in cervical cancer.Methods The databases of Pubmed,Embase,Science Direct and Cochrane Library were searched to collect all the literatures of the prognostic correlation of SCC-Ag for cervical cancer and evaluate the quality of the included literatures according to the evaluation criteria.Hazard ratio (HR) and 95% confidence interval (CI) were used to evaluate the correlation between SCC-Ag and prognosis of cervical cancer,and the subgroup analysis was also performed.the forest map and funnel map were plotted out by StataSE12.0 software.Results A total of 14 articles met to inclusion criteria were included.The high expression of SCC-Ag correlated significantly with the poor overall survival,(OS) in prognosis of cervical cancer (HR =2.73,95% CI =1.48-5.05,P =0.001).The disease free survival (DFS) of low expression of SCC-Ag was longer than that of high expression of SCC-Ag (HR =2.17,95% CI =1.84-2.57,P <0.01).No correlation of high expression of SCC-Ag with progression-free survival (PFS) of the patients was found (HR =2.68,95% CI =0.99-7.24,P =0.051).There was a significant heterogeneity of prospective OS (I2 =93.1%,P <0.01) in the subgroup analysis,but there was no statistical heterogeneity of prognostic DFS in the subgroup analysis (12 =23%,P =0.239).Conclusion SCC-Ag should be an important prognostic factor for cervical cancer.High expression of SCC-Ag may associate with poor prognosis of cervical cancer with statistical significance.
5.MicroRNA expression based tumor origin classification
Fesler ANDREW ; Hua LIU ; xiang Shi GUO ; fang Jing JU
Chinese Journal of Clinical Laboratory Science 2017;35(11):801-803
Approximately 3 to 5% of newly diagnosed metastatic cancers are of unknown primary tissue origin due to difficulties identifying a primary tumor using standard diagnostic approaches.MicroRNAs (miRNAs) have recently been demonstrated to be able to assist pathologist with improved accuracy in diagnosing cancers of unknown primary origin (CUP).In this short commentary,we will highlight some of the recent advancements in miRNA based cancer diagnosis as well as some future directions for the field.
6.Clinical manifestation and laboratory examinations of invasive Scedosporium infection
hua Li SHUAI ; hua Long HU ; ping He XU ; hong Chang RAO ; xun Xiao HU ; Jian HUANG
Chinese Journal of Clinical Laboratory Science 2017;35(11):872-876
Objective To investigate the clinical manifestation and the characteristics of laboratory examinations of invasive Scedocporium infection.Methods The clinical data of 8 patients infected with Scedosporium from January 2011 to April 2017 were collected and retrospective analysis combined with related literatures was performed.Results Among the 8 patients,6 strains of S.apiospermum,1 strains of Peudallescheria boydii and 1 strains of S.prolificans were detectable.The predisposing factors of Scedosporium infection were trauma,environmental exposure and hypoimmunity.The septahypha in specimens could be direcdy observed under microscopic examination with positive rate 100%.The growth speed of cultured colony was relatively fast and the invasiveness was strong.The colony of Scedosporium displayed various forms from white cashmere to black yeast sample.The color was gradually become dark from the center of colony with lengthening time of cultivation.Scedosporium could be identified by microscopic morphology combining culture technique.Conclusion The course of invasive Scedosporium infection may progress rapidly with serious and dangerous illness state.The most common infection of Scedosporium should be induced by S.apiospernum.The knowledge and understanding for Scedosporium infection should be strengthened to improve the level of diagnosis and treatment.
7.Establishment of allele-specific PCR for the detection of CYP3A5 and MDR-1 gene polymorphisms and its clinical application
Jie WU ; zhan Jia HOU ; Qian CHENG ; yao Jia LI ; dan Dan LI ; cong Yi YIN ; Wei SU
Chinese Journal of Clinical Laboratory Science 2017;35(11):844-848
Objective To establish an allele-specific PCR method for the detection of cytochrome P-450 CYP3A5 (A6986G) and multiding resistance gene MDR-1 (C3435T) polymorphisms,and investigate the correlations of their polymorphisms with blood tacrolimus (Tac) concentration/dose (C/D) ratio in renal transplant recipients.Methods The allele-specific PCR primers were designed according to the polymorphism sites of CYP3A5 (A6986G) and MDR-1 (C3435T) genes.Then,their polymorphisms in the genomic DNA of peripheral blood samples from 72 renal transplant recipients were analyzed,and the results were validated by DNA sequencing.The blood Tac concentration was determined by the chemiluminescence microparticle immunoassay and the differences of concentration,dose and C/D ratio of blood Tac in renal transplant recipients with different genotypes were compared at 1 month after transplantation.Results The coincidence rate between the established allele-specific PCR and DNA sequencing was 100%.The frequencies of CYP3A5 * 1/* 1,* 1/* 3 and * 3/* 3 genotypes in 72 renal transplant recipients were 18.1%,31.9% and 50.0%,respectively,and those of MDR-1 C/C,C/T and T/T genotypes were 27.8%,58.3% and 13.9%,respectively.There were significant differences in blood Tac concentration (P =0.014) and Tac C/D ratio (P =0.019) between different CYP3A5 genotypes of renal transplant recipients.Further analysis found that the Tac C/D ratio of CYP3A5 * 3/* 3 genotype was significantly higher than that of CYP3A5 * 1/* 1 and * 1/* 3 genotypes (P < 0.05).Conclusion The allele-specific PCR method for the detection of CYP3A5 and MDR-1 polymorphisms is successfully established and the polymorphism of CYP3A5 * 3 gene in renal transplant recipients is obviously correlated with the pharmacokinetics of Tac.
8.Verifying performance characteristics of the SLCO1B1 & APOE gene polymorphism detection kit
Yuan LV ; Fan QIU ; Xue XUE ; Yang YU ; xin Tong DU ; zheng Zi WANG ; Feng WANG
Chinese Journal of Clinical Laboratory Science 2017;35(11):868-871
Objective To verify the performance characteristics of the human SLCO1B1 & APOE gene detection kit (PCR-fluorescent probe technique).Methods The verification protocol for the human SLCO1B1 & APOE gene detection kit manufactured by Wuhan YZY Medical Science and Technology Co.,Ltd.was designed by referring to the requirements of professional standard and pertinent literatures,and its accuracy,specificity,lowest detection limit and anti-interference ability were compared with those of sequencing technique (gold standard).Results The results coincidence of 20 clinical samples from fluorescence PCR and sequencing technique was 100%,which met the requirement of accuracy.Forty clinical samples with wild type loci determined by fluorescence PCR were verified by sequencing and no any mutation was detected,which met the requirement of specificity.The lowest detection limit of the kit was 10 ng/μL.Three interfering substances,including hemoglobin,bilirubin and triglyceride,were individually added into the blood samples with known genotype,and the results determined by the kit were coincident with that without interfering substances,which met the requirement of anti-interference ability.Conclusion The verification parameters of the human SLCO1B1 & APOE gene detection kit are basically consistent with the manufacturer's statement,indicating that the kit meets the requirements of the relevant quality management and may be applied to the detection of clinical samples.
9.Molecular epidemiological characteristics and genotype analysis of norovirus in Zhenjiang region
feng Jin XU ; Hong XU ; sheng Yin SONG ; Yuan XUE ; he Shi SHAO ; xing Hong SHEN
Chinese Journal of Clinical Laboratory Science 2017;35(11):857-861
Objective To analyze the molecular epidemiological characteristics and genotype distribution of norovirus in Zhenjiang region during 2015 and 2016.Methods The fecal samples from diarrhea patients in Zhenjiang region were collected,and the expression level of norovirus capsid protein gene was detected by reverse transcription PCR (RT-PCR).The PCR products were validated by gene sequencing.Then,phylogenetic tree was constructed with Mega software,and sequence evolution and genotype were analyzed.Results The positive rate of norovirus infection was 5.30% (85/1 605).Among them,the detection rates of G Ⅰ genogroup and G Ⅱ genogroup were 0.87% (14/1 605) and 4.55% (73/1 605),respectively,and 2 samples were detected both of G Ⅰ and G Ⅱ genogroups.The highest positive rate of G Ⅱ genogroup occurred in adolescent patients aged from 10 to 20 years (17.80%).During 2015,the detection rate of norovirus infection was the highest in January,and then in November and February.During 2016,the detection rate of norovirus infection was the highest in December,and then in March and April.The main genotypes of norovirus in 2015 and 2016 were new G Ⅱ.17 variant (63.89%) and G Ⅱ.4 Sydney_2012 strain (35 %),respectively.Conclusion The infection of norovirus in Zhenjiang region appears G Ⅱ as main genogroup,adolescent as main patients,November to April of the next year as main epidemic seasons,and G Ⅱ.17 variant and G Ⅱ.4 Sydney_2012 strain as main epidemic genotypes.
10.Association of interleukin-6 gene-174G > C polymorphisman with coronary heart disease of Chinese Han population in Nanjing area
bang Yue WANG ; xia Hong WEI ; ye Zhi XU ; Yan WANG ; Kui ZHANG
Chinese Journal of Clinical Laboratory Science 2017;35(11):853-856
Objective To investigate the relationship of-174G > C polymorphism of interleukin 6 (IL-6) gene promoter region upstream-with coronary heart disease (CHD) of Chinese Han population in Nanjing area.Methods The polymorphism of IL-6-174G >C gene was confirmed in 235 patients with CHD and 175 healthy individuals by PCR-RFLP,and Hardy-Weinberg equilibrium was tested.DNA samples were selected for sequencing to verify their genotype.The concentration of blood sugar,lipid,C reaction protein (CRP) and glycosylated hemoglobin (HbAlc) in the samples were measured simultaneously.Results The genotype distributions of GG,GC and CC were 98.7%,1.3% and 0 in CHD group and 97.7%,2.3% and 0 in control group,respectively.The frequencies of G and C alleles were 99.4%,0.6% and 98.9%,1.1% in the two groups.There were no statistical significance for frequencies of genotype and alleles between the two groups (all P > 0.05).Compared with the control group,the differences of smoking,systolic blood pressure (SBP),diastohc blood pressure (DBP),triglyceride (TG),cholesterol (CHOL),low-density lipoproteins-cholesterol (LDL-C),high-density lipoproteins-cholesterol (HDL-C),apolipoprotein A-I (ApoA-I),ApoB,CRP and HbA1 c were statistically significant (all P < 0.05),while age,sex and blood glucose were not statistically significant (all P > 0.05).Conclusion IL-6-174 G > C gene polymorphism should not remarkably correlated with CHD in Chinese Han population in Nanjing area.