Table of Content

30 July 2022, Volume 37 Issue 7

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Expression of lncRNA HULC in papillary thyroid carcinoma tissues

JI Ye, YUAN Xiaosun, ZHANG Lei, MA Huili, XUE Yongfei, LI Changsheng, ZHANG Jingwei, REN Zhonghai, ZHANG Tengfei

2022, 37(7):  605-609.  DOI: 10.3969/j.issn.1673-8640.2022.07.001

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Objective To investigate the expression of long non-coding RNA（lncRNA）highly up-regulated in liver cancer（HULC） in papillary thyroid carcinoma（PTC） tissues,and to study the effect of interfering with this gene on cell proliferation activity and invasiveness. Methods A total of 105 patients with PTC who underwent surgical treatment were enrolled. The PTC tissues removed during the operation and the adjacent tissues more than 2 cm from the tumor edge were collected. The relative expression level of HULC was determined by real-time fluorescence quantitative polymerase chain reaction（RT-qPCR）. The TPC-1 cells were transfected with different sequences. According to the different transfection sequences of TPC-1 cells,they were classified into si-HULC group（transfected with small molecule interference sequence of HULC）,si-Con group（transfected with negative control sequence） and blank group（not transfected）. The expression of HULC,cell proliferation activity and invasiveness in each group were determined. Results The relative expression level of HULC in PTC tissues was higher than that in adjacent tissues（P<0.001）. The differences of the expression levels of HULC in PTC tissues with different tumor diameters,TNM stages and lymph node metastasis had statistical significance（P<0.05）. There was no statistical significance for the expression levels of HULC in PTC tissues with different ages,sex,the numbers of lesions and the types of local invasion（P>0.05）. The relative expression level of HULC,cell proliferation activity and the number of invasive cells in si-HULC group were lower than those in si-Con group and blank group（P<0.05）,and there was no statistical significance between si-Con group and blank group（P>0.05）. Conclusions The expression of HULC in PTC tissues is increased,and it is related to tumor size,TNM stage and lymph node metastasis. Interfering with HULC expression in TPC-1 cells can inhibit cell proliferation activity and invasiveness.

Clinical characteristic and genetic variation analysis of compound heterozygous mutation of BCHE gene leading to neonatal butyrylcholinesterase deficiency

WANG Juan, YU Jing, CHEN Jun, ZHENG Hong, DAI Liying

2022, 37(7):  610-614.  DOI: 10.3969/j.issn.1673-8640.2022.07.002

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Objective To investigate the clinical characteristic and genetic variation analysis of 2 cases of neonatal butyrylcholinesterase deficiency（BCHED） caused by compound heterozygous mutation of BCHE gene. Methods Totally,the clinical characteristics of 2 cases of BCHED were analyzed,and trio-based whole-exome sequencing was performed. The harm of variants was evaluated by bioinformatics analysis,which were verified by Sanger sequencing. Results The 2 cases had decreased serum cholinesterase and increased bilirubin,without liver function injure. Case 1 had neonatal diarrhea symptom,and Case 2 had neonatal jaundice symptom. The results of genetic analysis showed that there were compound heterozygous mutations（c.401_c.402insA and c.221delC; c.401_c.402insA and c.127G>A） in BCHE gene in the 2 cases. Sanger sequencing confirmed the existence of mutation. c.221delC and c.127G>A were not reported in ClinVar and HGMD databases,which were new mutations. Conclusions The gene variation and phenotype spectra have further expanded through the discovery of different phenotypes and new variations of BCHE gene in the 2 neonatal cases of BCHED.

Influence of specimen state on the accuracy of plasma Epstein-Barr virus DNA determination results

WANG Xiaoyu, SEMIRE Ainiwaer, LI Shijun

2022, 37(7):  615-617.  DOI: 10.3969/j.issn.1673-8640.2022.07.003

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Objective To investigate the expression of Epstein-Barr virus（EBV）DNA in hemolysis and lipoidemia of plasma samples. Methods A total of 40 plasma positive samples with confirmed EBV DNA were collected and divided into 2 groups randomly（20 cases for each group）. The EBV DNA positive samples were mixed with sterilized distilled water in a 1∶1 ratio as control samples. The levels of EBV DNA in simulated hemolysis（hemoglobin concentration 60.50,30.25 and 15.13 g/L） and simulated lipoidemia（triglyceride concentration 6.36,4.24 and 2.12 mmol/L） were determined by fluorescence quantitative polymerase chain reaction（PCR）,and statistical analysis was performed. Results When the hemoglobin concentrations were 60.50 and 30.25 g/L,the PCR results of EBV DNA positive plasma hemolysis samples and control samples showed statistical significance（P<0.05）. When the hemoglobin concentration was 15.13 g/L,there was no statistical significance in the PCR results between EBV DNA positive plasma hemolysis samples and control samples（P>0.05）. There was no statistical significance in the PCR results between lipoidemia samples and control samples（P>0.05）. Conclusions Hemolysis samples with hemoglobin concentration ≤15.13 g/L and lipoidemia samples with triglyceride concentration ≤6.36 mmol/L had no effect on the determination of EBV DNA plasma samples.

Relationship of fasting C-peptide with nonalcoholic fatty liver disease in children with obesity

AO Jiajia, ZHANG Junfeng, NIE Wenjian, YE Qing

2022, 37(7):  618-622.  DOI: 10.3969/j.issn.1673-8640.2022.07.004

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Objective To investigate the relationship of fasting C-peptide with nonalcoholic fatty liver disease（NAFLD） in children with obesity. Methods A total of 211 obese children diagnosed with NAFLD by abdominal ultrasound（NAFLD group） were enrolled,and 119 obese children were enrolled as control group. Among them,serum levels of total protein（TP）,albumin（Alb）,gamma glutamyltransferase（GGT）,alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,total cholesterol（TC）,triglyceride（TG）,creatinine（Cr）,fasting blood glucose（FBG）,high-density lipoprotein cholesterol（HDL-C）,low-density lipoprotein cholesterol（LDL-C）,white blood cell（WBC） count,platelet（PLT） count,hemoglobin（Hb）,fasting C-peptide,glycated hemoglobin A_1c（HbA_1c）,fasting insulin（FINS） were determined,and homeostasis model assessment for insulin resistance（HOMA-IR） was calculated. Using propensity score matching,82 patients were matched in a ratio of 1∶1 with sex,age,blood pressure,body mass index（BMI） as control variables and the relation of fasting C-peptide and the risk of NAFLD were analyzed. Multivariate stepwise Logistic regression analysis was used to evaluate the risk factors of NAFLD. Results After propensity score matching,the levels of ALT,AST,GGT,TP,fasting C-peptide,FINS and HOMA-IR in NAFLD group were higher than those in control group（P<0.05）,and there was no statistical significance in the other indicators between the 2 groups（P>0.05）. Multivariate stepwise Logistic regression analysis showed that ALT and fasting C-peptide were risk factors for NAFLD [odds ratios（OR） were 1.057 and 3.307,95% confidence intervals（CI） 1.036-1.078 and 1.439-7.602,respectively]. According to the tertiles of fasting C-peptide,the children were classified into C-peptide≤1.06 pmol/mL group,C-peptide 1.07-1.41 pmol/mL group and C-peptide ≥1.42 pmol/mL group. After the adjustment for confounding factors（age,sex,BMI,waist circumference,systolic blood pressure,ALT,Cr,TG,HDL-C,HOMA-IR and WBC count）,C-peptide ≥1.42 pmol/mL was a risk factor for NAFLD（OR= 4.635,95%CI 1.610-13.346）. Conclusions Fasting C-peptide plays a role in the early diagnosis of NAFLD in children with obesity.

Relationship between Vasostatin-2 and the occurrence of chronic heart failure and its prognosis in patients with myocardial infarction

PAN Wenqi, LU Lin

2022, 37(7):  623-627.  DOI: 10.3969/j.issn.1673-8640.2022.07.005

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Objective To study the relationship between the serum levels of Vasostatin-2 and chronic heart failure（CHF） in patients with myocardial infarction and its prognosis. Methods A total of 450 patients with CHF after myocardial infarction were enrolled as CHF group,and the general clinical data,including age,sex,hypertension,diabetes mellitus,renal function,blood lipid,fasting blood glucose,glycated hemoglobin A_1c（HbA_1c） and N-terminal pro-B-type natriuretic peptide（NT-proBNP）,were collected. Serum Vasostatin-2,tumor necrosis factor-alpha（TNF-α） and high-sensitivity C-reactive protein（hs-CRP） levels were determined. A total of 149 healthy subjects with negative results of coronary angiography were enrolled as control group. All the CHF patients were followed up for 3 years to record major adverse cardiovascular events（MACE）,and they were classified into MACE and MACE-free groups according to whether MACE occurred. The correlations between the indicators were assessed by Pearson correlation analysis or Spearman correlation analysis. Logistic stepwise regression analysis was used to assess the risk factors for developing MACE in CHF patients. Results Serum Vasostatin-2 levels were lower in CHF group than those in control group（P<0.001）,and serum TNF-α,hs-CRP and NT-proBNP levels were higher than those in control group（P<0.001）. There was statistical significance for serum Vasostatin-2 levels between different stages in CHF groups（P<0.001）. Correlation analysis results showed that Vasostatin-2 levels were positively correlated with left ventricular ejection fraction（LVEF）（r=0.377,P<0.05）,and they were negatively correlated with NT-proBNP,TNF-α,hs-CRP and heart failure stages（r values were -0.294,-0.267,-0.312 and -0.288,P<0.05,respectively）. Compared with MACE-free group,NT-proBNP,hs-CRP and TNF-α were increased in MACE group（P<0.001）,and Vasostatin-2 was decreased（P<0.001）. Vasostatin-2,NT-proBNP,hs-CRP,TNF-α,advanced age,hypertension,diabetes mellitus,renal function,severe heart failure stags（C and D） and LVEF were all risk factors for MACE in CHF patients（P<0.05）. Conclusions Serum Vasostatin-2 level is correlated with the occurrence and prognosis of CHF after myocardial infarction and can be used as one of the indicators of adjuvant diagnosis and prognostic judgment in CHF patients.

Relationship of 25（OH）D₃ and liver inflammatory stage with the efficacy of antiviral therapy

LIU Yang, GENG Kunjing, LI Hongjiang, SHI Haoxi, CHEN Sisi

2022, 37(7):  628-631.  DOI: 10.3969/j.issn.1673-8640.2022.07.006

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Objective To investigate the relationship between serum 25-hydroxyvitamin D₃ [25（OH）D₃] levels and the efficacy of antiviral therapy and liver inflammatory stage in patients with chronic hepatitis B（CHB）. Methods Totally,120 patients with CHB were enrolled and classified into auxiliary group（65 cases） and conventional group（55 cases）. They were classified into stage G1 group（12 cases）,stage G2 group（54 cases）,stage G3 group（39 cases） and stage G4 group（15 cases） according to the Scheuer scores of liver inflammatory stages. Pearson or Spearman correlation analysis was used to assess the correlation between the indicators. Results The differences in the rates of hepatitis B virus（HBV） DNA and hepatitis B e antigen（HBeAg） conversion between conventional and auxiliary groups at 12 weeks of treatment had no statistical significance（P>0.05）,and the differences in the rates of HBV DNA and HBeAg conversion at 24 weeks of treatment were statistically significant（P<0.05）. The difference in serum 25（OH）D₃ levels between auxiliary and conventional groups before treatment was not statistically significant（P>0.05）. After 24 weeks of treatment,serum 25（OH）D₃ levels in the 2 groups were higher than those before treatment（P<0.01）,and those of auxiliary group were higher than those in conventional group（P<0.001）. Serum interferon-gamma（INF-γ）,interleukin-6（IL-6） and tumor necrosis factor-alpha（TNF-α） levels were increased with the increasing liver inflammatory stages in CHB patients（P<0.05）,and serum 25（OH）D₃ levels were decreased with the increasing liver inflammatory stages（P<0.05）. Spearman correlation analysis showed that 25（OH）D₃ was negatively correlated with liver inflammatory stage（r=-0.715,P<0.001）. Pearson correlation analysis showed that 25（OH）D₃ was negatively correlated with IL-6 and TNF-α（r=-0.262 and -0.216,P<0.05） and had no correlation with INF-γ（r=-0.053,P>0.05）. Conclusions Serum 25（OH）D₃ levels in CHB patients are related with liver inflammatory stage and antiviral therapy efficacy.

Expression and clinical significance of SIRT-1 and CD44 in ovarian cancer tissues

LIU Rong, WANG Degang

2022, 37(7):  632-6358.  DOI: 10.3969/j.issn.1673-8640.2022.07.007

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Objective To investigate the expressions and clinical significance of silence information regulator-1（SIRT-1） and CD44 in ovarian cancer tissues. Methods The cancer tissues and adjacent tissues with positive resection margin >1 cm from 116 patients with ovarian cancer were collected,and the expressions of SIRT-1 and CD44 were determined by immunohistochemistry. The clinicopathological data of all the patients were collected. The correlation between SIRT-1 and CD44 was assessed by Spearman rank correlation analysis. Results The expression positive rates of SIRT-1 and CD44 in ovarian cancer tissues were higher than those in adjacent tissues（P<0.05）. In the expression positive rates of SIRT-1 and CD44 in ovarian cancer tissues,there was no statistical significance in the different ages,disease types and tumor diameters（P>0.05）,those of histological grade G1 were higher than those of G2-G3,those of the International Federation of Gynecology and Obstetrics（FIGO） stage Ⅲ - Ⅳ were higher than those of Ⅰ - Ⅱ,those of the infiltration degree T3-T4 were higher than those of T1-T2,and those with lymph node metastasis were higher than those without lymph node metastasis（P<0.05）. Spearman grade correlation analysis showed that SIRT-1 was positively correlated with CD44 expression in ovarian cancer tissues（r=0.721,P<0.001）. Conclusions SIRT-1 and CD44 are related to the occurrence and development of ovarian cancer,or they can be the markers for the disease severity asseessment of ovarian cancer patients.

Relationship of sCD147,sCD40L and miR-21 with carotid plaque type and prognosis in ACI patients

U Yuping, CHEN Yang

2022, 37(7):  636-640.  DOI: 10.3969/j.issn.1673-8640.2022.07.008

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Objective To investigate the relationship of soluble CD147（sCD147）,soluble CD40 ligand（sCD40L） and microRNA-21（miR-21） with carotid plaque type and prognosis in patients with acute cerebral infarction（ACI）. Methods Totally,159 patients with ACI were enrolled. According to the modified Rankin Scale（mRS） score after 3 months,they were classified into poor prognosis group（47 cases） and good prognosis group（112 cases）. The baseline data,carotid plaque types,the National Institute of Health Stroke Scale（NIHSS） scores and serum sCD147,sCD40L and miR-21 levels were compared between the 2 groups,and the correlations of the above indicators with carotid plaque types and NIHSS scores were analyzed by Spearman or Pearson correlation analysis. The adverse factors affecting the prognosis of ACI and the predictive values of the above indicators on the prognosis of ACI were evaluated by multiple linear regression analysis. Results Serum sCD147,sCD40L and miR-21 levels were higher in poor prognosis group than those in good prognosis group（P<0.05）. Serum sCD147,sCD40L and miR-21 were positively correlated with stable plaque,unstable plaque and NIHSS score（P<0.05）. Multiple linear regression analysis showed that serum sCD147,sCD40L and miR-21 were correlated with poor prognosis（mRS score） in ACI after adjusting age,carotid plaque type,NIHSS score,abnormal lipid metabolism and time from onset to thrombolysis（P<0.05）. Conclusions There is a correlation between serum sCD147,sCD40L,miR-21 and carotid plaque types in ACI patients,which can be used for prognosis evaluation in ACI patients.

Establishment of the reference intervals for related parameters of adult reticulocytes in Shanghai

XU Qianqian, JIANG Wangqing, SHEN Linjie, LU Yingjie, CHEN Jian, JIANG Haoqin

2022, 37(7):  641-645.  DOI: 10.3969/j.issn.1673-8640.2022.07.009

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Objective To establish the reference intervals for adult reticulocyte（RET）-related parameters in Shanghai. Methods The RET-related parameters [the absolute value of reticulocyte（RET#）,the percentage of reticulocyte（RET%）,high fluorescence intensity reticulocyte percentage（HFR%）,medium fluorescence intensity reticulocyte percentage（MFR%）,low fluorescence intensity reticulocyte percentage（LFR%）,immature reticulocyte fraction percentage（IRF%）,reticulocyte hemoglobin（RET-He） and reticulocyte production index（RPI）] of 2 080 healthy adults were determined. According to sex,the research subjects were classified into male and female groups,and according to ages,they were classified into 18-40-year-old,41-60-year-old and 61-80-year-old groups. Another 40 samples for blood routine test were collected for the verification of reference intervals. Results The results of normality test showed that the RET-related parameters were skewed distribution（P<0.05）. There was no statistical significance in RET%,RET# and RPI among different age groups（P>0.05）. There was statistical significance in IRF%,LFR%,MFR%,HFR% and RET-He between 41-60-year-old group,61-80-year-old group and 18-40-year-old group（P<0.05）,while there was no statistical significance between 41-60-year-old and 61-80-year-old groups（P>0.05）. The reference intervals were established by percentile method after combining age and sex groups. The reference intervals of RET%,RET# and RPI in males were 0.92%-2.15%,（46.30-107.30）×10⁹/L,0.9-2.4,and those in females were 0.87%-2.27%,（37.96-101.90）×10⁹/L,0.6-1.8,respectively. The reference intervals of IRF%,LFR%,MFR% and HFR% in 18-40-year-old males were 4.1%-16.1%,83.9%-96.0%,3.6%-12.1%,0.2%-5.0%,those in 18-40-year-old females were 3.9%-17.9%,82.1%-96.1%,3.6%-12.5%,0.1%-5.5%,and those in 41-80-year-old males and females were 4.4%-18.0%,82.0%-95.6%,4.0%-12.8%,0.2%-5.1%,respectively. The reference interval of RET-He in 18-40-year-old males was 30.3-35.5 pg,that in 18-40-year-old females was 27.6-34.6 pg,that in 41-80-year-old males was 30.4-35.7 pg,and that in 41-80-year-old females was 27.8-34.6 pg. The results of 40 verification samples were all in the reference intervals. Conclusions The reference intervals of RET-related parameters for adults in Shanghai have been established preliminarily,which can improve the clinical application roles of RET-related parameters.

Application of polymerase chain reaction melting curve method in determining hypertensive drug-related gene mutations

WAN Changchun, YANG Hui, ZHUANG Xuewei

2022, 37(7):  646-651.  DOI: 10.3969/j.issn.1673-8640.2022.07.010

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Objective To assess the clinical application of polymerase chain reaction（PCR） melting curve method in determining hypertensive drug-related gene mutations. Methods Totally,100 hypertension patients were enrolled,and the single nucleotide polymorphism（SNP） sites of hypertensive drug-related gene mutations,including cytochrome P450 2D6（CYP2D6）rs1065852,cytochrome P450 2C9（CYP2C9）rs1057910,betal-adrenergic receptor-1（ADRB1）rs1801253,angiotensin Ⅱ receptor 1（AGTR1）rs5186 and angiotensin-converting enzyme（ACE）rs1799752,were determined. DNA sequencing was performed. The accuracy,repeatability,anti-interference and the genotype frequency of PCR melting curve method were evaluated. Results The consistency of PCR melting curve method and DNA sequencing was 100%. The extremum values（R） of the Tm about 5 gene sites were all <0.5 ℃. Hemoglobin（Hb）≤2 g/L,bilirubin（Bil）≤342 μmol/L,triglyceride（TG）≤37 mmol/L had no effect on PCR melting curve method. The coefficient of variation（CV）of Tm was ≤5%. In 100 hypertension patients,there was 1 case of AGTR1（c.1166A>C） and 1 case of CYP2C9*3（c.1075A>C）homozygous mutations. The genotype frequencies of CYP2D6*10 and CYP2C9*3 were 39.5% and 5.5%,respectively. The genotype frequencies of C allele of mutant ADRB1,C allele of mutant AGTR1 and D allele of ACE deletion were 62.0%,5.0% and 35.0%,respectively. Conclusions PCR melting curve method is simple and has low requirements for equipments,and it can be used as a routine method for determining anti-hypertensive drug-related gene mutations.

UroQuick rapid urine culture technique for urinary tract infection in children

PAN Fen, PAN Yuxuan, SHI Yingying, YU Fangyuan, ZHANG Hong

2022, 37(7):  652-656.  DOI: 10.3969/j.issn.1673-8640.2022.07.011

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Objective To investigate the role of UroQuick rapid urine culture technique for urinary tract infection（UTI） in children. Methods A total of 235 clean midstream urine samples were collected from children with suspected UTI aged 0-14 years from December 2019 to January 2020. Conventional urine culture and UroQuick rapid urine culture were performed simultaneously. Taking the results of conventional urine culture as gold standard,the diagnostic efficacy of UroQuick rapid urine culture for UTI was further evaluated. Results Among the 235 samples,58（24.7%） samples,including 43 samples of bacteria,4 samples of fungi and 11 samples of bacteria and fungi,were positive by conventional urine culture,while 69 samples,including 55 samples of bacteria,3 samples of fungi and 11 samples of bacteria and fungi,were positive by UroQuick rapid urine culture. The consistency rate of the 2 methods was 86.8%（204/235）. Compared to conventional urine culture,the sensitivity,specificity,positive predictive value and negative predictive value of turbidity value of UroQuick rapid urine culture were 84.48%,88.70%,71.01% and 94.58%,respectively. Furthermore,the sensitivity,specificity,positive predictive value and negative predictive value of bacterial concentration of UroQuick rapid urine culture were 82.76%,92.66%,78.69% and 94.25%,respectively. Conclusions UroQuick rapid urine culture is a new technique for urine culture and has high specificity and negative predictive value whether in turbidity value or in bacterial concentration,which will provide a reference for the early screening of UTI in children.

Expression and bioinformatics analysis of hsa-miR-34a in tumors

WEN Shuzhan, FU Zile, CHEN Shuying

2022, 37(7):  657-663.  DOI: 10.3969/j.issn.1673-8640.2022.07.012

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Objective To analyze the expression of homo sapiens microRNA（miR）-34a（hsa-miR-34a） in different tumors,to predict its target genes,and to analyze its biological function and mechanism. Methods Sequence was obtained from miRBase database to analyze the position of hsa-miR-34a in human genome and its conservation among species. TargetScan,miRWalk,miRDB and mirDIP were used to predict the target gene set of hsa-miR-34a and take the intersection,the expression of which in different human tissues was analyzed through TiGER database. Gene Ontology（GO） enrichment analysis and REACTOME signaling pathway enrichment analysis of hsa-miR-34a target genes were performed using PANTHER and DAVID online tools. The interaction network of hsa-miR-34a target genes corresponding to target proteins was constructed by STRING database. Results The expressions of hsa-miR-34a in breast cancer,prostate cancer,bladder cancer and other tumors were decreased. The sequence encoding hsa-miR-34a was highly conserved among different species. A total of 225 target genes were obtained,and the top 30 target genes were selected after Target Score functional sequencing. There was no statistical significance in the relative expression of target genes among different human tissues（P>0.05）. GO enrichment analysis showed that hsa-miR-34a target genes were mainly expressed in clathrin vesicles,vesicle membranes and their material transport,multicellular biological processes and development,neurogenesis and development,anatomical structure development and so on. REACTOME signaling pathway enrichment analysis showed that hsa-miR-34a target genes were concentrated in biological signaling pathways such as botulinum toxin B and G toxicities,cell adhesion and migration inhibition mediated by semaphorin 4D（Sema4D） and the synthesis and release of neurotransmitters. The target genes were reordered according to the protein interaction network. The target genes corresponding to the top 5 proteins were SYT1,VAMP2,MET,RRAS and CACNA1E. Conclusions The hsa-miR-34a is related to the occurrence and development of a variety of tumors. The analysis of its target genes and biological functions can provide a reference for subsequent clinical and basic research.

Key genes and signal pathways of Alzheimer's disease based on GEO database

HOU Yuli, WANG Yifei, FU Jingxuan, WANG Peichang

2022, 37(7):  664-668.  DOI: 10.3969/j.issn.1673-8640.2022.07.013

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Objective To screen the differentially expressed genes（DEG） and signal pathways related to Alzheimer's disease（AD） using bioinformatics analysis methods based on GEO database. Methods GSE118553 was selected from GEO database as the analysis dataset,and GSE106241 was used as the validation dataset of key genes. The DEG were selected from the GSE118553 dataset matrix. Using cluster profile package,Gene Ontology（GO） analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway analysis on DEG were preformed. Protein-protein interaction（PPI） network was constructed to screen out the top 10 key genes involved in AD. GSE106241 dataset was used to verify the expression differences of 10 key genes in temporal cortex tissues of AD patients with different braak grades to healthy controls and the correlation with amyloid bata-protein（Aβ）42 expression. Results There were 157 DEG obtained in GSE118553 dataset,including 88 up-regulated genes and 69 down-regulated genes. GO analysis and KEGG pathway analysis showed that DEG were enriched in gamma-aminobutyric acid（GABA） signaling pathway,neurotransmitter transmission and synaptic transmission and so on. A PPI network of DEG was constructed,and 10 key genes（SNAP25,SYT1,GRIN2A,SLC12A5,GAD1,GABRG2,GABRD,PVALB,GABRB2 and FN1） related to Alzheimer's disease were verified. The GSE106241 dataset was used for verification,and the results showed that there was statistical significance in the expression of SNAP25,SYT1,GRIN2A,SLC12A5,GAD1,GABRG2,GABRD,PVALB and GABRB2 in temporal cortex tissues of AD patients with different braak grades（P<0.05）. Pearson correlation analysis showed that the expression of SNAP25,SYT1 and SLC12A5 were negatively correlated with the expression of Aβ42（r values were -0.354,-0.283 and -0.310,respectively,P<0.05）. Conclusions The obtained key genes,SNAP25,SYT1 and SLC12A5,may be potential biomarkers of Alzheimer's disease and provide new targets for the diagnosis and treatment of AD.

Status of quality management of new laboratories for nucleic acid determination of SARS-CoV-2 in Zhejiang

CHEN Qian, SHAN Zhiming, SONG Chao, KANG Fengfeng, JIN Jing, LI Weixing

2022, 37(7):  669-673.  DOI: 10.3969/j.issn.1673-8640.2022.07.014

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Objective To investigate the status of quality management of new severe acute respiratory syndrome coronavirus 2（SARS-CoV-2） nucleic acid determination laboratories constructed after January 2020 in Zhejiang. Methods A total of 179 SARS-CoV-2 nucleic acid determination laboratories,including 107 new laboratories and 72 laboratories constructed before January 2020（original laboratories）,were enrolled randomly to on-site inspection and sample assessment. The statistical analysis of non-conforming terms and assessment results was performed. Results The non-conforming terms of new laboratories were more than original laboratories,and the problems from new laboratories and original laboratories were mainly internal quality control and performance verification. The determination system matching rate of new laboratories was 25.23%,and that of original laboratories was 75.00%. The false negative results of routine testing and rapid testing were all from new laboratories. Conclusions Internal quality control and performance verification are common problems to all the laboratories. It should strengthen the monitoring of weak positive internal quality control cycle threshold（Ct） values and implement proper performance verification protocols. There is a quality gap between new laboratories and original laboratories. It should strengthen the quality management of rapid testing and strengthen personnel skill training.

Evaluation of uncertainty of serum total bilirubin in calibration curve method by Monte Carlo method

MENG Shuting, ZHOU Weiping, DAI Huifen, DONG Deping, DONG Yefeng, GE Renmei, WANG Huimin

2022, 37(7):  674-679.  DOI: 10.3969/j.issn.1673-8640.2022.07.015

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Objective To evaluate the uncertainty process for total bilirubin reference measurement procedure which was recommended by the Joint Committee for Traceability in Laboratory Medicine（JCTLM） used by the Monte Carlo method（MCM） through optimizing. To realize quantity traceability about the uncertainty evaluation of serum total bilirubin under reference laboratory conditions. Methods Applying Doumas method which was recommended by JCTLM,sample A and sample B from external quality assessment scheme for reference laboratories of the International Federation of Clinical Chemistry and Laboratory Medicine（IFCC） in 2014 were determined. Based on software MATLAB,the uncertainty of total bilirubin determination was evaluated by MCM. Results The uncertainty model which was evaluated by the Guide to the Expression of Uncertainty in Measurement（GUM）was established. Parameters related to the calibration curve were as follows：slope（b） was 0.007 531 6,the standard deviation of slope was 0.000 025 51,mean concentration was 87.284 4 mg/L,the standard deviation of mean concentration was 0.196 5,mean absorbance（$\bar{y}$） was 0.657 917,the standard deviation of mean absorbance was 0.000 365 4,intercept（a） was 0.000 525 16,and the standard deviation of intercept was 0.002 699. The results of total bilirubin determination under laboratory conditions were as follows：the concentration of sample A x was 39.368 8 mg/L,the standard uncertainty of sample A synthesis（u） was 0.603 2,95% confidence interval（CI） was 38.188 0-40.553 5,the concentration of sample B x was 18.703 1 mg/L,the standard uncertainty of sample B synthesis（u）was 0.296 4,and 95%CI was 18.122 5-19.284 4. Output quantity of MCM presented skewed normal distribution,showing peak fat-tail. The GUM uncertainty framework（GUF） inclusion interval was verified with adaptive MCM. The absolute deviation d_low and d_high values of the 2 groups were greater than the numerical tolerance 0.05. The results of GUF did not pass the MCM verification. Conclusions With the help of visual software MATLAB,MCM can optimize calculation process,improve the accuracy of inspection results highly compared with the classical GUF.

Research on performance specification of thrombelastogram

MIAO Yingbo, ZHAO Qiang, SONG Ying, ZHU Peichao, ZHOU Wei, XU Chong

2022, 37(7):  680-683.  DOI: 10.3969/j.issn.1673-8640.2022.07.016

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Objective To investigate how to obtain the appropriate evaluation standard based on the change or constant concentration value as performance standard through thrombelastogram proficiency verification data,and to improve continuously and effectively the determination level of thrombelastogram. Methods The data reported by laboratories participating in the 2018-2020 proficiency verification of Shanghai Center for Clinical Laboratory for thrombelastogram were collected. The relation of U-scores between the reported data of coagulation reaction time,coagulation formation time,angle and maximum amplitude of thrombus and the target value was evaluated,and the evaluation standard of each index based on the change or constant concentration value was obtained. Results The constant concentration values of coagulation reaction time,angle and maximum amplitude of thrombus could be used as evaluation criteria. Coagulation formation time can adopt the evaluation standard based on the change of concentration value. Conclusions The performance specifications of each index of thrombelastogram obtained in this study can be applied to the evaluation standards of proficiency verification and the requirements of allowable imprecision of internal quality control,which can evaluate the determination level of thrombelastogram in Shanghai objectively.

Overview of detection methods for Mycoplasma in cell preparations

CHEN Naihan, SA Yalian

2022, 37(7):  684-687.  DOI: 10.3969/j.issn.1673-8640.2022.07.017

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Mycoplasma is a common exogenous contaminant during cell culturing. There are many detection methods for Mycoplasma contamination. However,due to the fact that the final product of living cell preparations can not be sterilized,the detection method of Mycoplasma is culturing and indicator cell culturing（DNA staining method）,and the detection time is long. The rapid release test of the final cell preparation has certain limitation,so it is necessary to improve the Mycoplasma detection methods for the final cell preparation. This review briefly introduces the morphology,culturing and nucleic acid detection method of Mycoplasma,hoping to provide a reference for the construction of the evaluation system of cell preparation quality monitoring technology.

Influence factors of blood culture negative results and progress of pathogen detection in bloodstream infection

CHEN Hanlu, WU Shenghai

2022, 37(7):  688-694.  DOI: 10.3969/j.issn.1673-8640.2022.07.018

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Bloodstream infections refer to disseminated infections caused by pathogenic microorganisms invading bloodstream,including bacteremia and septicemia,which can lead to serious clinical consequence and high fatality rates. As the gold standard for the diagnosis of bloodstream infection,blood culture has high specificity,but an unsatisfactory positive rate and considerable patients with bloodstream infection have negative blood culture results. On one hand,the using of antibiotics before blood collection,insufficient blood volume,extended pre-analysis time and other factors should be accountable. On the other hand,there are some fastidious microorganisms needing prolonged incubation. The diagnostic efficiency of bloodstream infection can be improved by the enrichment of pathogens after reporting positive in culture flasks and rapid identification with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF MS） and fluorescence in situ hybridization（FISH）. Serological test and molecular biotechnology,such as serum antibody determination,bacterium 16S rRNA,real-time fluorescence polymerase chain reaction（PCR）,multiplex PCR and next-generation sequencing（NGS）,can improve the detection rate of pathogens in bloodstream. This review focuses on the influence factors of negative blood culture and the progress of laboratory determination of bloodstream infection.