Table of Content

20 April 2017, Volume 32 Issue 4

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Orginal Article

Value and risk of next generation sequencing in the diagnosis and treatment of cancer

FAN Qishi, WU Beiying

2017, 32(4):  245-249.  DOI: 10.3969/j.issn.1673-8640.2017.04.001

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Sequencing technology has been developed from the 1st generation to the 4th one,and the length of reads is from long to short and from short to long again. Its progress has promoted the development of life science and clinical medical research. Next generation sequencing （NGS） technology has high flux,high accuracy and huge quantity of information. Different sequencing platforms have their own advantages in flux,length of reads,accuracy,speed and cost. They play important roles in de novo genome sequencing,resequencing,transcriptome and apparent genetics. In recent years,the datum output of NGS shows the trend of exponential growth,and has been gradually applied to clinical services,such as personalized medicine and genetic diagnosis. NGS in clinical oncology research and determination also shows a huge market value,but at the same time it hides certain degree of risk,which should be paid great attention and vigilance.

Application and prospect of high-throughput sequencing in clinical molecular diagnosis

JIANG Xiaofeng

2017, 32(4):  250-254.  DOI: 10.3969/j.issn.1673-8640.2017.04.002

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：High-throughput sequencing,next generation sequencing（NGS）,is a new type of genetic screening technology. Its innovation accelerates the understanding of genetic markers and molecular mechanism,and also promotes clinical gene diagnosis from single gene to multi-genes,especially for complex genetic diseases. As the new sequencing technology used in clinic,doctors will change the management of genetic diseases,and patients will be placed into different groups eventually based on genetic diagnosis. In addition,with the improvement of bioinformatics and package,the application of NGS in clinic will be more routine. This review discusses the major platform and performance of NGS and its application in clinic and the process of NGS analysis of sequence variations.

Next generation sequencing in quality control and analysis of data

YU Dong, GUO Yingjun

2017, 32(4):  255-261.  DOI: 10.3969/j.issn.1673-8640.2017.04.003

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With the development of precision medicine in China,next generation sequencing （NGS） has been improved in clinical practice. Though NGS is a new technology in the field of clinical laboratory medicine,there is a few systematic laws,regulations and guidelines to regulate the development of NGS. Changhai Hospital is one of the first national trial units,which is applying NGS to the diagnosis and therapy of cancer. Now,it has conducted 2 NGS programs,which are genetic testing program and cancer individualized medication program. This review will focus on the problems from practice and provide a useful reference.

Reliability of m.3700G>A mutation as screening locus for mitochondrial disease

XU Bing, FU Qingzi, YANG Yanling, SHEN Lijun

2017, 32(4):  262-266.  DOI: 10.3969/j.issn.1673-8640.2017.04.004

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Objective To evaluate the role of nicotinamide-adenine dinudeotid-dehydrogenase subunit 1 （ND1）gene m.3700G>A mutation for mitochondrial disease,especially Leber's hereditary optic neuropathy. Methods Mitochondrial adenosine triphosphate（ATP） generation,respiratory chain complex activity and steady state level and the level of reactive oxygen species （ROS） were determined in cybrids with m.3700G and m.3700A. Results Mitochondrial oxidative phosphorylation complex Ⅰ was not affected in cells with mutation of m.3700G>A. Mitochondrial ATP generation and the level of ROS were not affected as well. Conclusions The mutation of m.3700G>A might not be related with mitochondrial disease. Therefore,the screening of m.3700G>A mutation is not advised.

Polymorphism of ALDH2 gene with allele specific fluorescence polymerase chain reaction

SUN Yujing, XIAN Haipeng, LIU Xiangyi, LIU Chang, LONG Yan, SUN Yuanyuan, ZHAO Xiaotao

2017, 32(4):  267-271.  DOI: 10.3969/j.issn.1673-8640.2017.04.005

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Objective To investigate the distribution of acetaldehyde dehydrogenase 2 （ALDH2） gene polymorphism,and to evaluate the feasibility of allele specific fluorescence polymerase chain reaction （PCR）for ALDH2 gene polymorphism. Methods Allele specific fluorescence PCR and DNA sequencing were used for ALDH2 genotyping in peripheral blood of 375 patients. Gene chip method was adopted to determine the specimens with inconsistent results by the 2 methods. Results Among the 375 specimens,248 cases （66.1%） of ALDH2*1/ 1,107 cases （28.5%） of ALDH2*1/ 2 and 20 cases （5.4%） of ALDH2*2/ 2 were determined by allele specific fluorescence PCR,while 246 cases （65.6%） of ALDH2*1/ 1,108 cases （28.8%） of ALDH2*1/ 2 and 21 cases （5.6%） of ALDH2*2/ 2 were determined by DNA sequencing,respectively. There was no statistical significance for ALDH2 genotyping between allele specific fluorescence PCR and DNA sequencing （χ²=1.33,P=0.392）,with good consistency（κ=0.978,P<0.001）. The coincidence rate was 98.9% （371/375）between the 2 methods. For 4 cases with inconsistent results,the results of gene chip method was in agreement with those by allele specific fluorescence PCR. Conclusions Allele specific fluorescence PCR for ALDH2 genotyping is feasible,and could meet the requirements of clinical ALDH2 gene determination.

Whole exome sequencing for identifying HPS1 as causative gene in a pedigree with albinism

YOU Guoling, ZHANG Lichen, ZHANG Xiaoqing, LI Xiaoliang, FU Qihua, WANG Bo

2017, 32(4):  272-275.  DOI: 10.3969/j.issn.1673-8640.2017.04.006

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Objective In this study,whole exome sequencing was used to identify disease-causing genes for a pedigree with autosomal recessive albinism and pathogenesis mechanism. Methods Genomic DNA was extracted from peripheral blood samples of proband and other family members. Whole exome sequencing was performed for proband. The disease-causing mutations were determined by whole exome sequencing with sequence variation bioinformatics analysis,and the mutations were identified by Sanger sequencing. Results Proband was diagnosed with compound heterozygous mutations in HPS1 gene,including c.1276_1279dupGGAG （p.Asp427fs） mutation and c.398+5G>A mutation. The frameshifting mutation was inherited from father,and the splicing mutation was inherited from mother. Conclusions The compound heterozygous c.1276_1279dupGGAG and c.398+5G>A mutations of HPS1 gene are identified as causative mutations for albinism in this family. The proband is diagnosed as Hermansky-Pudlak syndrome 1 subtype. The whole exome sequencing can be a new and efficient method to determine specific disease gene and subtype in albinism,and it can improve the treatment and patients' live quality.

Levels of serum vitamin A,25-hydroxyvitamin D and vitamin E of children under 6 years old in Jiamusi

WANG Limin, ZHANG Xueling, WANG Wenjuan, WANG Haiyan, HUO Mingxia, YUAN Xiaohui, YUAN Boyang

2017, 32(4):  276-279.  DOI: 10.3969/j.issn.1673-8640.2017.04.007

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Objective To investigate the levels of serum vitamin A（Vit A）,25-hydroxyvitamin D [25（OH）D] and vitamin E（Vit E） of children under 6 years old in Jiamusi,and to provide a reference for the reasonable supplementation of Vit A,vitamin D（Vit D） and Vit E and preventing from related diseases. Methods According to ages,5 169 children were classified into 6 groups,<1-year-old group（732 cases）,1-year-old group（831 cases）,2-year-old group （693 cases）,3-year-old group（1 164 cases）,4-year-old group （1 008 cases） and 5-year-old group（741 cases）. After achieving questionnaires,the venous blood was extracted. The levels of Vit A and Vit E were determined by high-performance liquid chromatography,and the level of 25（OH）D was determined by high-performance liquid chromatography tandem mass spectrometry. Results The level of Vit A was（0.28±0.087）mg/L. The insufficiency rate was 54.09%,and the deficiency rate was 13.46%. There was statistical significance with different ages （F=4.370,P=0.001）. The minimal level of Vit A was （0.25±0.071）mg/L in <1-year-old group,the insufficiency rate was 61.48%,and the deficiency rate was 19.67%. The level of 25（OH）D was （24.20±9.97）ng/mL,the insufficiency rate was 18.57%,and the deficiency rate was 17.99%. With the increasing of ages,the levels of 25（OH）D decreased. The average level of Vit E was（8.68±2.49）mg/L,the insufficiency rate was 21.53%,and the deficiency rate was 0.58%. With the increasing of ages,the level of Vit E had a decreasing tendency. Conclusions The nutritional status of Vit A and 25（OH）D of children under 6 years old is not optimistic in Jiamusi. It is necessary to determine and give reasonable supplementation of vitamin for high-risk subjects in order to prevent from vitamin deficiency disorder. The nutritional status of Vit E is relatively good.

Preliminary investigation for the reference intervals of serum 25-hydroxyvitamin D among minors in Shanghai

GAO Zhengfeng, XU Menghua, YUAN Meifen, XU Jin

2017, 32(4):  280-283.  DOI: 10.3969/j.issn.1673-8640.2017.04.008

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Objective To establish the reference intervals of serum 25-hydroxyvitamin D [25（OH）D] among minors in Shanghai. Methods A total of 400 minors （≤16 years old） were enrolled,including 200 boys and 200 girls. They were classified into 5 groups according to ages,<1 month（20 cases）,1-<6 months（80 cases）,6 months-<3 years （100 cases）,3-<6 years（100 cases）and 6-16 years（100 cases）. The level of 25（OH）D was determined by enzyme-linked immunosorbent assay （ELISA）. Results In the results of 400 cases,1 outlier was eliminated. The 25（OH）D levels demonstrated skewed distribution,so its overall level was shown by median （M）[quartiles（P₂₅-P₇₅）]. The 25（OH）D level of 399 cases was 55.64（42.93-76.05） nmol/L,56.47（44.59-76.48） nmol/L for boys and 55.44（41.07-75.55） nmol/L for girls. There was no statistical significance between boys and girls（P>0.05）. There was no statistical significance for different ages （P>0.05）. The 25（OH）D level in the groups of 1-<6 months and 6 months-<3 years were higher than those in the other 3 groups（P<0.05）. The reference intervals should be established according to ages. The reference intervals were shown by percentiles [the 2.5th percentile（P_2.5）-the 97.5th percentile （P_97.5）] and were 32.66-79.33 nmol/L for <1 month group,32.45-129.91 nmol/L for 1-<6 months group,38.98-129.90 nmol/L for 6 months-<3 years group,32.22-86.44 nmol/L for 3-<6 years group and 30.74-87.74 nmol/L for 6-16 years group. Conclusions The reference intervals of 25（OH）D have been initially established for minors in Shanghai.

Correlations of cognitive impairment with vitamin B₁₂ and folic acid in elderly dementia patients

HU Yao, GUO Qihao, CEN Yi, LIN Yao

2017, 32(4):  284-289.  DOI: 10.3969/j.issn.1673-8640.2017.04.009

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Objective To investigate the correlations of vitamin B₁₂ and folic acid with mild cognitive impairment （MCI） and Alzheimer's disease（AD）. Methods Serum vitamin B₁₂ and folic acid levels were determined by chemiluminescence microparticle immunoassay in 191 elderly dementia patients ≥ 50 years old and 69 cognitive normal subjects（normal control group）. The low levels of vitamin B₁₂ and folic acid as high-risk factors for AD were analyzed. Cognitive function was evaluated with comprehensive neuropsychological tests,such as Chinese Version of Mini Mental State Examination（CMMSE） and the Memory and Executive Screening（MES）. Pearson's correlation analysis was performed to evaluate the relationship among serum vitamin B₁₂,folic acid,age,education year,body mass index（BMI） and cognitive function in all groups. Logistic regression was used to evaluate the association of vitamin B₁₂ and folic acid with MCI and AD. Results A total of 54 patients with MCI and 137 patients with AD were determined. There were 78.1% patients with normal vitamin B₁₂ and folic acid levels,6.1% patients with low vitamin B₁₂ level,5.8% patients with low folic acid level,and 10.0% with high vitamin B₁₂ or folic acid level. Vitamin B₁₂ or folic acid level had no correlation with CMMSE and MES scores after covariates' adjustments（P>0.05）. Low vitamin B₁₂ level was correlated with the risk of AD [odds ratio（OR）=2.869,95% confidence interval（CI）1.123-7.333]. Conclusions Vitamin B₁₂ and folic acid are not correlated with cognitive impairment. However,low vitamin B₁₂ level may be correlate with an increased risk for AD.

Food intolerance testing for recurrent aphthous ulcer

WANG Guoqing, WANG Cuihuan, ZHANG Lili, TONG Xin, LIU Chenlu, SHANG Jianwei

2017, 32(4):  290-294.  DOI: 10.3969/j.issn.1673-8640.2017.04.010

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Objective To investigate food intolerance testing for the treatment and prevention of recurrent aphthous ulcer （RAU）. Methods Serum samples of 410 patients with RAU and 120 healthy subjects （healthy control group）were collected. Serum food-specific immunoglobulin （Ig）G antibodies against 14 foods were determined by enzyme-linked immunosorbent assay. Results A total of 350 cases in RAU group（85.37%） were determined food-specific IgG antibodies positive,and the positive rate was higher than that in healthy control group（28.33%,P<0.01）. Furthermore,the positive rate of multiple food intolerance in RAU group was higher than that in healthy control group （P<0.01）. In RAU group,the positive rate and level of egg-specific IgG antibody were the highest. The levels of food-specific IgG antibodies in RAU group were higher than those in healthy control group （P<0.01）. RAU group showed a mild intolerance to beef,shrimp,chicken and pork,and the positive rates of food-specific IgG antibodies against them had no statistical significance between RAU and healthy control groups（P>0.05）,while the levels of food-specific IgG antibodies in RAU group were higher than those in healthy control group （P<0.01）. The food-specific IgG antibody level of IgG positive in RAU group was higher than that of IgG negative in RAU group （P<0.01）,and the food-specific IgG antibody level of IgG negative in RAU group was higher than that in healthy control group （P<0.01）. After a 3-month follow-up of RAU group,a modification of dietary brought an effective relief to ulcer in 258 of 406 cases （63.55%）. Conclusions There is a close relationship between food intolerance and RAU,and the identification of intolerable foods and avoiding consumption of them are of value in clinical treatment and prevention of RAU.

Influence of red blood cell transfusion with radiation on serum inflammatory factors and prognosis of perioperative patients with esophageal carcinoma

LIU Ye, ZHOU Ye, CHEN Bo, TANG Xiaofeng

2017, 32(4):  295-298.  DOI: 10.3969/j.issn.1673-8640.2017.04.011

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Objective To evaluate the influence of red blood cell transfusion with radiation on serum inflammatory factors and prognosis of perioperative patients with esophageal carcinoma. Methods A total of 159 patients with esophageal carcinoma were enrolled. All patients were randomly classified into radiation-free group （78 cases）and radiation group （81 cases）. The radiation-free group was given variant suspended red blood cell. The radiation group was given variant suspended red blood cell with radiation （¹³⁷Cs,25 Gy）. The levels of interleukin （IL）-6,IL-10,tumor necrosis factor alpha（TNF-α）and interferon gamma（IFN-γ）were determined before and after surgery in the 2 groups. The prognosis was evaluated. Results On the 1st and 7th days after surgery,the levels of IL-6 and IL-10 in radiation-free group were higher than those before surgery （P<0.05）,and were higher than those in corresponding radiation group （P<0.05）. On the 1st and 7th days after surgery,the level of IFN-γ in radiation-free group was lower than that before surgery （P<0.01）,and was lower than that in corresponding radiation group （P<0.01）. On the 1st day after surgery,the level of TNF-α in radiation-free group was lower than that before surgery （P<0.05）,and was lower than that in corresponding radiation group （P<0.05）. On the 1st and 7th days after surgery,the levels of indices mentioned above in radiation group had no statistical significance compared with those before surgery （P>0.05）. The days staying in hospital and in intensive care unit （ICU）had no statistical significance between the 2 groups （P>0.05）. For 1,3 and 5 years after surgery,the survival rates in radiation group were higher than those in radiation-free group （P<0.05）. Conclusions Compared with common blood transfusion,perioperative blood transfusion with radiation in patients with esophageal carcinoma has no immune suppression,and has a good prognosis. Giving the complexity for the influence of blood transfusion on prognosis,large clinical trials are still needed to confirm the conclusions based on the control of other factors.

Distribution and drug resistance of 17 361 isolates from blood culturing in a certain region

WANG Xin, BAI Yuanyuan, WANG Cuicui, SHEN Cuihua, JIN Yan

2017, 32(4):  299-303.  DOI: 10.3969/j.issn.1673-8640.2017.04.012

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Objective To analyze the distribution and drug resistance of isolates from blood culturing in Shandong,and to provide a reference for the treatment of bloodstream infection. Methods The isolates from blood culturing were collected and analyzed retrospectively from January 2015 to December 2015 in Shandong. The results were assessed according to the Clinical and Laboratory Standards Institute （CLSI）2014 standard,and WHONET 5.6 software was used for drug resistance analysis. Results In the 17 361 isolates,Gram-negative bacilli accounted for 54.9% （9 524 isolates）,and its isolation rate was higher than that of Gram-positive cocci（accounting for 45.1%,7 837 isolates）. The top 5 isolates were Escherichia coli,coagulase-negative Staphylococcus,Klebsiella pneumoniae,Staphylococcus aureus and Pseudomonas aeruginosa. The isolation rates of extended-spectrum beta-lactamases （ESBLs）-producing Escherichia coli and Klebsiella pneumoniae were 53.5% and 25.8%,respectively. The drug resistance rates of ESBLs-producing isolates to carbapenems,oxygenase inhibitors and cephalosporin were lower. The drug resistance rates to carbapenems in Pseudomonas aeruginosa and Acinetobacter baumannii were high,and carbapenems should be chosen to use carefully. The determination rate of methicillin-resistant Staphylococcus aureus （MRSA） was 27.1%,which was less than that of methicillin-resistant coagulase-negative Staphylococcus （MRSCNS,80.2%）. No Staphylococcus resistant to vancomycin and linezolid was found. The drug resistance rates of Enterococcus faecium and Enterococcus faecalis to vancomycin were 1.7% and 1.2%,and those to linezolid were 0.7% and 4.7%. Vancomycin can be still as the first choice for Gram-positive cocci in bloodstream infection. Conclusions The analysis on the distribution and drug resistance of isolates from blood culturing could guide the rational use of medicines in clinic and reduce the mortality in severe infection.

Correlations of red blood cell distribution width and related inflammation factors with disease activity in patients with rheumatoid arthritis

LI Yunchun, ZHONG Li, WANG Yue, FANG Zhongjun, DING Liumei, SU Qizhu

2017, 32(4):  304-307.  DOI: 10.3969/j.issn.1673-8640.2017.04.013

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Objective To investigate the correlations of red blood cell distribution width （RDW） and related inflammation factors with disease activity in patients with rheumatoid arthritis （RA）. Methods A total of 110 patients with RA,50 patients with gout and 50 healthy subjects were enrolled. RDW,erythrocyte sedimentation rate （ESR）,C-reactive protein（CRP）,rheumatoid factor（RF） and anti-cyclic citrullinated peptide（CCP） antibody were determined. According to disease activity score（DAS）28 levels,RA patients were classified into 2 groups,high-active group（DAS 28>5.1,60 cases） and low-active group （DAS 28 2.6-3.2,50 cases）. Results RDW,CRP,ESR,anti-CCP antibody and RF were increased gradually in healthy control,gout,RA high-active and low-active groups （P<0.01）. The rate of bone erosion in patients with RA was 60.9%. RDW,ESR and CRP in patients with bone erosion was higher than those in patients without bone erosion （P<0.01）. High titers of anti-CCP antibody （>300 RU/mL） and RF （>200 IU/mL） in RA group were also related with high bone erosion rates. Conclusions RDW and high titers of anti-CCP antibody and RF can be used as parameters for evaluating disease activity of RA.

Two enzyme methods for determining drug concentrations in biochemical instruments

LI Jiayong, LIU Chang, WANG Hongming, ZHANG Rulin

2017, 32(4):  311-315.  DOI: 10.3969/j.issn.1673-8640.2017.04.015

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Objective To evaluate the consistency of homogeneous enzyme immunoassay and enzyme-amplified immunoassay in the determinations of phenytoin,valproic acid,theophylline,digoxin,methotrexate,carbamazepine and mycophenolic acid in serum and cyclosporine A in whole blood with 8 drug concentrations. Methods Homogeneous enzyme immunoassay and enzyme-amplified immunoassay were used to determine BIO-RAD with 3 concentrations and original quality control products,and the within-run and between-run precisions were calculated. A total of 120 clinical samples were collected for each drug concentration,which more than 30% of them were abnormal. Except for cyclosporine A,30 homologous plasma samples were collected for each drug concentration. Using SIEMENS ADVIA 2400 automatic biochemical analyzer,homogeneous enzyme immunoassay and enzyme-amplified immunoassay were performed parallelly,and the consistency of them was evaluated. Results The within-run and between-run precisions of homogeneous enzyme immunoassay and enzyme-amplified immunoassay were good,and there was a good stability. The overall correlation of each drug concentration was good,and r were 0.997 for cyclosporine A,0.993 for phenytoin,0.997 for valproic acid,0.996 for theophylline,0.988 for digoxin,0.994 for methotrexate,0.999 for carbamazepine and 0.997 for mycophenolic acid. All the regression equations were significant statistically （P<0.001）. There was no statistical significance for the results of the 2 methods（P>0.05）. The expected bias of each drug concentration at medical decision level was in allowable error range. The consistency of homologous serum and plasma samples was good,except for cyclosporine A,and each regression equations were significant statistically（P<0.01）,and the difference of paired t-test was not significant statistically （P>0.05）. Conclusions The results of phenytoin,valproic acid,theophylline,digoxin,methotrexate,carbamazepine,mycophenolic acid and cyclosporine A with 8 drug concentrations have good consistency between the 2 methods by SIEMENS ADVIA 2400 automatic biochemical analyzer.

Correlation of systemic lupus erythematosus clinical phenotypes with anti-double-stranded DNA antibodies and performance comparison of 4 commercial determination kits

SONG Rui, YE Ping, WEI Chaohui, CHEN Xiaoxiang, WANG Jiucun

2017, 32(4):  316-321.  DOI: 10.3969/j.issn.1673-8640.2017.04.016

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Objective To compare the performance of 4 commercial determination kits for anti-double-stranded DNA （dsDNA）antibodies,to analyze the correlation of 15 typical systemic lupus erythematosus （SLE）clinical phenotypes with anti-dsDNA antibodies,and to provide a reference for the diagnosis and monitoring of SLE. Methods A total of 229 SLE patients were enrolled,and 112 patients with other connective tissue diseases were enrolled. Their serum samples were collected. There were 10 typical SLE clinical phenotypes and 5 parameters. All SLE patients were evaluated according to SLE disease activity index （SLEDAI）. The levels of anti-dsDNA antibodies were determined by 4 commercial determination kits [kit A using indirect immunofluorescence assay（IFA）,kit B and kit C using enzyme-linked immunosorbent assay（ELISA）and kit D using radioimmunoassay]. The performance of the 4 commercial determination kits was evaluated. The correlation of typical SLE clinical phenotypes and parameters with anti-dsDNA antibodies was analyzed. Results The results of 4 commercial determination kits had statistical significance（P<0.01）. Kit C had the highest Youden's index,its Youden's index was higher than those of kit B and kit D,and that of kit A was the lowest. The results of kit B,C and D had correlations with SLEDAI （r= 0.482,0.512 and 0.685,P<0.01）. For 10 typical SLE clinical phenotypes and 5 parameters,anti-dsDNA antibodies were correlated positively with increasing anti-nucleosome antibody and decreasing complement （P<0.01）. The results of positive anti-nuclear antibodies （P<0.05）,nephropathy （P<0.05）,leukopenia （P<0.05） and malar rash（P<0.05） in 2 kits were positively correlated with anti-dsDNA antibodies. The other 8 typical SLE clinical phenotypes,including thrombocytopenia and peripheral arthritis,showed no correlation with anti-dsDNA antibodies. Conclusions The performance of 4 commercial determination kits is different. Kit C is adapted to screen SLE,and kit D is suitable for monitoring the activity of SLE. Nephropathy,malar rash,anti-nucleosome antibody,anti-nuclear antibody,decreasing complement and leukopenia are correlated with anti-dsDNA antibodies.

Cervista HR-HPV DNA testing combined with TCT in cervical cancer screening

ZHAO Peiyu, CHEN Aozheng, QIN Jinlong, SONG Liwen, CHENG Jiajing

2017, 32(4):  322-325.  DOI: 10.3969/j.issn.1673-8640.2017.04.017

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Objective To evaluate Cervista high-risk human papillomavirus （HR-HPV）DNA testing combined with thin cytology test （TCT）in cervical cancer screening. Methods A total of 745 female patients were diagnosed as precancerous lesions or cervical cancer with pathological diagnosis as gold standard. The performance of TCT,Cervista HR-HPV DNA testing and Cervista HR-HPV DNA testing combined with TCT were evaluated. The role of Cervista HR-HPV DNA testing combined with TCT in cervical cancer screening was evaluated. Results The specificity of TCT was the best（50.40%）. The sensitivity and negative predictive value of Cervista HR-HPV DNA testing combined with TCT were 92.51% and 82.31%,which were higher than those of single determinations. Conclusions The determination rate of Cervista HR-HPV DNA testing is higher than that of TCT,the combined determination can improve the sensitivity and negative predictive valve effectively,and can decrease the rate of missed diagnosis and false positive rate.

MALDI-TOF MS combined with short-term culturing for determining pathogenic bacteria rapidly in midstream urine

ZHANG Jinghao, FANG Yi, ZHANG Yanmei, ZHAO Hu

2017, 32(4):  326-330.  DOI: 10.3969/j.issn.1673-8640.2017.04.018

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Objective To evaluate a method for determining pathogenic bacteria rapidly in midstream urine by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF MS）combined with short-term culturing. Methods A total of 1 873 samples of midstream urine were collected. Each sample was divided equally into 2 parts. One was used for routine midstream urine culturing and identified by Vitek 2 Compact,the other was used for short-term culturing for 4 or 6 h after centrifugation,and then moss was collected and identified directly by MALDI-TOF MS. Identification by Vitek 2 Compact and quantitative culturing was considered as gold standard to evaluate the sensitivity and specificity of MALDI-TOF MS combined with short-term culturing for determining pathogenic bacteria in midstream urine. Results A total of 381 samples were determined with pathogenic bacteria in 1 873 samples,and the positive rate was 20.3%. Among them,346 cases （90.8%） had single species,27 cases （7.1%） had 2 species,and 8 cases （2.1%）had≥3 species. In samples with single species,the determination rate in 4 h culturing group was 80.1% （277/346）,and the determination rate in 6 h culturing group was 97.1% （336/346） by MALDI-TOF MS directly. The coincidence rate of MALDI-TOF MS combined with 6 h culturing compared with routine culturing was 100.0%,except Acinetobacter baumannii,Enterococcus faecium and Enterococcus faecalis（their coincidence rates were 81.4%,76.9% and 86.7%）. Conclusions The midstream urine samples are cultured for 4-6 h after centrifugation,then identified by MALDI-TOF MS directly. This method could effectively shorten both culturing time and turn-around time.

Osteoprotegerin expression on aortic atherosclerotic plaques and vessel calcification of diabetes mellitus rats

SUN Yefu, CHEN Guiming, XIA Aiping, ZHANG Zhifeng

2017, 32(4):  333-337.  DOI: 10.3969/j.issn.1673-8640.2017.04.020

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Objective To investigate the expression of osteoprotegerin（OPG） on aortic atherosclerotic plaques and vascular calcification of diabetes mellitus rats and the interference of pioglitazone. Methods A total of healthy male SD rats were enrolled and classified into 6 groups randomly,and there were 7 rats in each group,including control group （group A）,diabetes mellitus group（group B）,aortic atherosclerotic plaque and vessel calcification group（group C）,diabetes mellitus+aortic atherosclerotic plaque and vessel calcification group（group D）,aortic atherosclerotic plaque and vessel calcification+pioglitazone group（group E） ,diabetes mellitus+aortic atherosclerotic plaque and vessel calcification+pioglitazone group（group F）. Blood was extracted from rats on the 30th day for the determinations of high-sensitivity C-reactive protein（hs-CRP）,fasting plasma glucose（FPG）,glycated hemoglobin A _1c（HbA_1c）and OPG. Calcium deposits in thoracic aorta and alkaline phosphatase（ALP） activity were determined. Results The success rate of diabetes mellitus rat modeling was 100%. Compared with group A,the levels of calcium deposits in thoracic aorta,ALP activities,OPG and hs-CRP in group C,D and E increased（P<0.05）. The levels of calcium deposits in thoracic aorta and ALP activities increased（P<0.05）,and there was no statistical significance for hs-CRP and OPG in group F（P>0.05）. The levels of calcium deposits in thoracic aorta,ALP activities and OPG had statistical significance between group C and E（P<0.05）. The levels of ALP activities,hs-CRP,FPG,HbA_1c and OPG between group D and F were statistically significant（P<0.05）. The levels of calcium deposits in thoracic aorta and OPG in group F were lower than those in group E（P<0.05）. OPG was positively related with the levels of calcium deposits in thoracic aorta,ALP activities and hs-CRP and FPG（r=0.654,0.218,0.539 and 0.711,P<0.05 or P<0.01）. Conclusions Pioglitazone can reduce the formation of aortic atherosclerotic plaques and vessel calcification,and the mechanism might be related to the regulation of OPG,the inhibition of inflammatory reaction and the decreasing of calcium salt deposition.

Identification of fibrinolytic enzyme-producing strain and optimization of producing enzyme conditions

LI Lizhuo, HUANG Huanjing, LI Zhuowei, ZHENG Mei

2017, 32(4):  338-342.  DOI: 10.3969/j.issn.1673-8640.2017.04.021

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Objective To identify fibrinolytic enzyme-producing strain （LZ-01） and optimize producing enzyme conditions. Methods The morphological characteristics of LZ-01 were observed. Physiological and biochemical tests and the sequence similarity analysis of phoR gene were adopted to determine the strain. Through single factor experiment,the influence of carbon source and nitrogen source on enzyme activity was investigated,and the optimal conditions of enzyme production were determined by orthogonal experiment. Results After the identification,the similarity of this strain and Bacillus amyloliquefaciens standard strain FZ42 was 99.78%. So,the strain was identified as Bacillus amyloliquefaciens. After optimizing,the producing enzyme activity increased by 3.63 times. Conclusions This study provides a theoretical basis for the further development of fibrinolytic drugs.

Preparation and performance evaluation of quality control material for homocysteine

SHI Jianxin, ZHU Yan, JIANG Ying, XIANG Yingjun, LIN Huiling, DUAN Guikai

2017, 32(4):  343-346.  DOI: 10.3969/j.issn.1673-8640.2017.04.022

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Objective To prepare a liquid internal quality control material for homocysteine （Hcy） without assigned value,and to investigate its performance. Methods Pooled human serum was used as the medium of the quality control material for Hcy．The homogeneity and stability of quality control material were evaluated according to CNAS-GL03. The self-made quality control material was also used in internal quality control and was compared with RANDOX quality control material. Results The homogeneity and stability of the self-made quality control material met related requirements．Compared with RANDOX quality control material,there was no statistical significance （P>0.05）. Conclusions The self-made quality control material for Hcy has good homogeneity and stability,and it can be used for the internal quality control in clinical laboratories.