Table of Content

30 May 2020, Volume 35 Issue 5

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Influence of common clinical interference factors on the detection of ctDNA EGFR T790M mutation

JIANG Wenrong, MIAO Yingxin, WANG Su, WANG Shiwen, ZHAO Hu, ZHANG Yanmei

2020, 35(5):  399-404.  DOI: 10.3969/j.issn.1673-8640.2020.05.001

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Objective To study the influence of common clinical interference factors on the detection of epidermal growth factor receptor（EGFR） T790M mutation of circulating tumor DNA（ctDNA） in plasma. Methods T790M wild-type plasma from 7 healthy donors was collected. Different concentrations of T790M mutant plasmid and certain interference factor（triglyceride,bilirubin and hemoglobin） were added into the T790M wild-type plasma to simulate interfered plasma. T790M mutation was detected by super amplification refractory mutation system（Super-ARMS） after DNA extraction. Each sample was detected for 3 times,the concentration and purity of DNA,Ct values of internal control and ΔCt values of T790M mutation were recorded. SPSS 22.0 software was used for statistical analysis. Results While the concentration of DNA of critically positive mutant samples could be affected by bilirubin（≥85.5 μmol/L） and triglyceride（≥9.25 mmol/L）（P<0.05）,the purity of DNA of them could not be affected. As low concentration of bilirubin（85.5 μmol/L） and high concentration of hemoglobin （2.0 g/L） affected the concentration of DNA of strongly positive mutant samples（P<0.05）,triglyceride （≥9.25 mmol/L） and hemoglobin（0.5-1.5 g/L） also affected the purity of DNA（P<0.05）. Lipemia had influence on the detection of T790M mutation by Super-ARMS,and triglyceride（≥9.25 mmol/L） affected the Ct values of internal control and ΔCt values of T790M mutation of both strongly and critically positive mutant samples（P<0.05）. Otherwise,high concentrations of bilirubin（≥256.5 μmol/L） affected the Ct values of internal control of strongly positive mutant samples,and middle concentrations of hemoglobin（≥1.0 g/L） affected their ΔCt values of T790M mutation（P<0.05）. Conclusions Different concentrations of hemoglobin,triglyceride and bilirubin have different effects on several indicators of detecting ctDNA EGFR T790M mutation by Super-ARMS. Plasma containing interference factors should be avoided in practical clinical detection as far as possible.

Influence of heating inactivation on the nucleic acid testing results of SARS-CoV-2 throat swab samples

LIU Jie, ZHAO Jianhong, GAO Yan, HAN Taoli, LI Li, KONG Yi, ZHAO Huanxing, SUN Lingli, WANG Chengbin

2020, 35(5):  405-408.  DOI: 10.3969/j.issn.1673-8640.2020.05.002

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Objective To investigate the influence of heating inactivation on the nucleic acid testing results of severe acute respiratory syndrome coronavirus 2（SARS-CoV-2） throat swab samples. Methods Totally,10 throat swab samples of positive SARS-CoV-2 were collected. The standard thermometer was used to calibrate the temperature and calculate temperature rising time. Throat swab samples of positive SARS-CoV-2 were classified into 56 ℃ 30 min treatment group and untreatment group. Nucleic acid was extracted and amplified by real-time fluorescence quantitative polymerase chain reaction（PCR）. Results It took 68 min to heat the sample to 56 ℃ in air bath and 11.5 min in water bath. The results of PCR in 56 ℃ 30 min treatment group after heating showed that the Ct value increased. Of the 10 samples,8 samples were determined as positive in 56 ℃ 30 min treatment group. The samples with low nucleic acid concentration cannot be determined after treatment. Conclusions The heating up time depends on heating medium. Heating inactivation will affect SARS-CoV-2 nucleic acid testing results.

Relationship between plasma small dense low-density lipoprotein and ischemic stroke

CHEN Yabin, CAI Hehui, ZHANG Jianming, SU Yongfa, ZHANG Zhishan

2020, 35(5):  409-413.  DOI: 10.3969/j.issn.1673-8640.2020.05.003

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Objective To investigate the role of plasma small dense low-density lipoprotein（sd-LDL） in ischemic stroke. Methods The levels of plasma lipoproteins,including very low-density lipoprotein（VLDL）,intermediate-density lipoprotein（IDL）,low-density lipoprotein（LDL）,large buoyant low-density lipoprotein（lb-LDL）,sd-LDL,high-density lipoprotein（HDL）,were determined in 181 patients with ischemic stroke（ischemic stroke group） and 40 healthy subjects （healthy control group）. The ratio of sd-LDL/LDL and the ratio of sd-LDL/HDL were calculated. The clinical data were collected,and the risk factors of ischemic stroke were evaluated by Logistic binary regression analysis. The role of each index in the diagnosis of ischemic stroke was evaluated by receiver operating characteristic（ROC） curve. Results Compared with healthy control group,the levels of plasma lb-LDL,sd-LDL,HDL,the ratio of sd-LDL/LDL and the ratio of sd-LDL/HDL in ischemic stroke group had statistical significance（P<0.05）. There was no statistical significance for the other indexes between the 2 groups（P>0.05）. Logistic binary regression analysis showed that systolic blood pressure,diastolic blood pressure,triglyceride,fasting blood glucose,lb-LDL,sd-LDL,HDL,the ratio of sd-LDL/LDL and the ratio of sd-LDL/HDL were the risk factors for ischemic stroke（P<0.05）. Systolic blood pressure was an independent risk factor for ischemic stroke [odds ratio（OR）=1.065,95% confidence interval（CI） 1.011-1.122）. ROC curve analysis showed that the areas under curves of sd-LDL,the ratio of sd-LDL/LDL and the ratio of sd-LDL/HDL were 0.675,0.741 and 0.762,respectively. The optimal cut-off values were 245 mg/L,0.225 and 0.342,respectively. The sensitivities were 58.18%,61.72% and 64.00%,and the specificities was 90.00%,75.00% and 70.00%,respectively. Conclusions Sd-LDL is related to the occurrence of ischemic stroke. The ratio of sd-LDL/LDL and the ratio of sd-LDL/HDL may be used in the diagnosis of ischemic stroke.

Relationship between alpha 2-Heremans-Schmid glycoprotein to adiponectin ratio and type 2 diabetes mellitus complicated with metabolic syndrome

ZHOU Zhongwei, JU Huixiang, JIN Hao, CHEN Hongmei, SUN Mingzhong

2020, 35(5):  414-419.  DOI: 10.3969/j.issn.1673-8640.2020.05.004

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Objective To investigate the relationship between alpha 2-Heremans-Schmid glycoprotein（AHSG） to adiponectin（APN） ratio and type 2 diabetes mellitus（T2DM） complicated with metabolic syndrome（MS）. Methods A total of 130 newly diagnosed patients with T2DM were enrolled,including 61 patients with only T2DM,69 patients with T2DM complicated with MS. A total of 60 healthy subjects were enrolled as control group. The general data of all the subjects were collected. Fasting plasma glucose,total cholesterol（TC）,triglyceride（TG）,high-density lipoprotein cholesterol（HDL-C）,low-density lipoprotein cholesterol（LDL-C）,C-reactive protein（CRP）,fasting insulin（FINS）,AHSG and APN levels were determined,and homeostasis model assessment of insulin resistance（HOMA-IR） index and AHSG/APN ratio was calculated. Spearman rank correlation analysis was used to analyze the relationships among various parameters. Logistic regression analysis was used to evaluate the risk factors of T2DM complicated with MS. Receiver operating characteristic（ROC） curve analysis was performed to evaluate the auxiliary diagnostic values of various parameters for T2DM complicated with MS. Results AHSG and AHSG/APN ratio increased gradually from control group,T2DM group to T2DM complicated with MS group,whereas APN decreased gradually among them（P<0.05）. Spearman rank correlation analysis showed that AHSG was positively correlated with waistline,fasting plasma glucose,TG,LDL-C,HOMA-IR,CRP and AHSG/APN ratio（P<0.01）,and it was negatively correlated with APN（P<0.01）. APN was negatively correlated with waistline,body mass index（BMI）,systolic and diastolic blood pressures,TC,AHSG and AHSG/APN ratio（P<0.05,P<0.01）,and it was positively correlated with HDL-C（P<0.05）. AHSG/APN ratio was correlated with all the parameters（P<0.05,P<0.01） except age（P>0.05）. Logistic regression analysis showed that only AHSG/APN ratio was the risk factor for MS in T2DM patients after adjusting for confounding factors [odds ratio（OR）=1.051,95% confidence interval（CI） 1.015-1.088,P<0.01],and APN was a protective factor of MS（OR=0.758,95%CI 0.566-0.978,P=0.047）. ROC curve analysis indicated the areas under curves（AUC） of AHSG,APN and AHSG/APN ratio in auxiliary diagnosis of MS were 0.692,0.730 and 0.806,respectively,and the diagnostic performance of AHSG/APN ratio was superior to either AHSG or APN. Conclusions AHSG/APN ratio would reflect the metabolic disorder and the risk of MS in T2DM patients,and it could have a auxiliary diagnostic value for T2DM complicated with MS.

Clinical application of neutrophil gelatinase-associated lipocalin in acute pancreatitis

GE Ling, WU Liying

2020, 35(5):  420-423.  DOI: 10.3969/j.issn.1673-8640.2020.05.005

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Objective To investigate the clinical application of neutrophil gelatinase-associated lipocalin（NGAL） in acute pancreatitis（AP）. Methods A total of 74 AP patients were enrolled,and there were 34 cases of mild acute pancreatitis（MAP） and 40 cases of severe acute pancreatitis（SAP）. A total of 20 healthy subjects were enrolled as healthy control group. Serum levels of NGAL,blood urea nitrogen（BUN）,creatinine（Cr）,C-reactive protein（CRP） and amylase（AMY） were determined. Using receiver operating characteristic（ROC） curve,the diagnostic performance of serum NGAL and AMY for AP was evaluated. Results Compared with SAP and MAP groups,serum NGAL,AMY and CRP levels were lower in healthy control group（P<0.05）,and NGAL and CRP levels in SAP group were higher than those in MAP group（P<0.05）. There was no statistical significance for AMY between the 2 groups（P>0.05）. BUN and Cr had no statistical significance among the 3 groups（P>0.05）. ROC curve analysis showed that the areas under curves（AUC） of NGAL and AMY for the diagnosis of AP were 0.722 and 0.784,the optimal cut-off values were 240 ng/mL and 250 U/L,the sensitivities were 90.0% and 83.3%,and the specificities were 55.4% and 63.9%,respectively. Conclusions Serum NGAL determination plays a role in the diagnosis of AP,which can reflect the degree of AP infection.

Role of PTX3 in the diagnosis of community-acquired pneumonia

ZHU Hong, ZHU Yuqing, GU Guobao

2020, 35(5):  424-427.  DOI: 10.3969/j.issn.1673-8640.2020.05.006

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Objective To evaluate the role of pentraxin-3（PTX3） in the diagnosis of community-acquired pneumonia（CAP）. Methods A total of 70 inpatients with CAP were enrolled,which were classified into severe CAP group and mild CAP group according to their condition. Totally,70 healthy subjects were enrolled as healthy control group. The results of C-reactive protein（CRP）,procalcitonin（PCT）,white blood cell（WBC） count and neutrophil/lymphocyte ratio（NLR） determinations were collected. Peripheral blood was collected within 24 h,on the 3rd day and the 7th day after admission,and serum PTX3 level was dynamically monitored. The correlation between different groups was compared,and the expression differences and dynamic changes of PTX3 were analyzed. Results PTX3,PCT and CRP in CAP group were higher than those in healthy control group（P<0.05）. Compared with mild CAP group,the increase of PTX3,PCT and CRP had statistical significance in severe CAP group（P<0.05）. The area under receiver operating characteristic（ROC） curve of PTX3 was 0.714,the sensitivity was 75.82%,and the specificity was 77.18%,when the cut-off value was 31.28 ng/mL. Dynamic monitoring of PTX3 showed that PTX3 level in severe CAP group was higher than that in mild CAP group（P<0.05）,and PTX3 levels in both groups gradually decreased with treatment time,but the PTX3 level in severe CAP patients remained at a high level on the 7th day. The main pathogen for CAP was bacteria,and there was no statistical significance in PTX3 level among the patients infected with different pathogens（P=0.296）. Conclusions Serum PTX3 level,especially the dynamic monitoring result,can be used as a biomarker for reflecting CAP,which can provide effective auxiliary diagnosis and efficacy monitoring for clinical practice.

Analysis of hepatitis B virus S protein mutation in chronic hepatitis B and cirrhosis patients

YUAN Yongming, ZHANG Xiaoying, GU Chao, ZHANG Jue, SUN Xuehua

2020, 35(5):  428-433.  DOI: 10.3969/j.issn.1673-8640.2020.05.007

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Objective To investigate the difference of hepatitis B virus（HBV） S protein mutations in patients with chronic hepatitis B （CHB） and cirrhosis,and to assess its role in clinical application. Methods A total of 114 patients with CHB were enrolled,and they were classified into 3 groups,asymptomatic HBV carriers（41 cases）,inactive hepatitis B surface antigen（HBsAg） carriers（38 cases） and cirrhosis patients（35 cases）. DNA sequencing was used to determine HBV S protein gene mutation. Results There was statistical significance in age and HBV DNA loads of asymptomatic HBV carrier,inactive HBsAg carrier and cirrhosis groups（P<0.01）,and there was no statistical significance in sex and HBV genotypes（P>0.05）. The gene mutation frequency of HBV S protein was higher in cirrhosis group than those in asymptomatic HBV carrier group（P<0.05） and inactive HBsAg carrier group（P<0.01）. The mutation rates in outside major hydrophilic region（MHR） and cytotoxic T lymphocytes（CTL） +helper T cell（Th） epitopes were increased in the 3 groups（asymptomatic HBV carrier<inactive HBsAg carrier<cirrhosis patient）（P<0.01）. There was no statistical significance among the 3 groups in MHR and a determinant,LOOP1,LOOP2 located in MHR and non-immune epitopes located outside MHR（P>0.05）. Single amino acid substitution was found in the majority of asymptomatic HBV carriers except 2 patients. Of these,one had 3 amino acid substitutions in CTL+Th epitopes and non-immune epitopes located outside MHR,and another had 2 amino acid substitutions in MHR and CTL+Th epitopes outside MHR. The mutation rates in CTL+Th epitopes and non-immune epitopes outside MHR were comparable and random for asymptomatic HBV carriers,whereas more than 2 mutation sites were determined in 5 cases of inactive HBsAg carriers（13.15%） and 9 patients with cirrhosis（25.71%）. These mutation sites preferentially concentrated at the CTL+Th epitopes outside MHR. Conclusions High prevalence of outside MHR variants especially CTL+Th epitopes is associated not only with hepatitis B e antigen（HBeAg）-negative serostatus but also with severe liver diseases,particularly cirrhosis.

Correlations of serum IL-1,IL-2,IL-6,IL-8,IL-10 and hs-CRP levels with neovascularization glaucoma

SONG Yunxiao, HUANG Xiaofeng, ZHANG Haichen, YAO Tianyue, YUAN Wenhua, ZHAO Zhiyun

2020, 35(5):  434-439.  DOI: 10.3969/j.issn.1673-8640.2020.05.008

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Objective To investigate the correlations of serum interleukin（IL）-1,IL-2,IL-6,IL-8,IL-10 and high-sensitivity C-reactive protein（hs-CRP） levels with neovascularization glaucoma（NVG）. Methods A total of 69 patients with NVG and 39 healthy subjects were enrolled. The levels of IL-1,IL-2,IL-6,IL-8,IL-10 and hs-CRP were determined. Ophthalmic examination [intraocular pressure（IOP）,central corneal thickness（CCT）,vertical cup-disc ratio（VCDR）,axial length（AL）,anterior chamber depth（ACD）,visual field mean defect（MD）,visual field mean sensitivity（MS） and so on] and physical examination were performed. There were 32 cases of retinal vein occlusion（RVO） secondary NVG and 37 cases of diabetic retinopathy（DR） secondary NVG. Spearman correlation analysis was used to evaluate the correlation between factors and ophthalmic parameters of NVG patients. Binary Logistic regression analysis was performed to identify the risk factors for NVG. Receiver operating characteristic（ROC） curve was used to analyze the value of each parameter in diagnosing NVG. Results Serum levels of IL-6,IL-8 and hs-CRP in NVG group were higher than those in control group（P<0.01）. There was no statistical significance in IL-1,IL-2 and IL-10 between the 2 groups（P>0.05）. There was no statistical significance in IL-1,IL-2,IL-6,IL-8,IL-10 and hs-CRP levels between DR secondary NVG group and RVO secondary NVG group（P>0.05）. Spearman correlation analysis showed that serum IL-6 level was positively correlated with MD in NVG patients（r=0.257,P<0.001）,and it was negatively correlated with MS（r=-0.196,P<0.001）. Serum IL-1,IL-2,IL-8,IL-10 and hs-CRP levels were not correlated with ophthalmic parameters（P>0.05）. Binary Logistic regression analysis showed that IL-6 [odds ratio（OR）=4.443,95% confidence interval（CI） 1.207-16.40],IL-8（OR=1.067,95%CI 1.001-1.237） and hs-CRP（OR=3.886,95%CI 1.191-12.683）were independent risk factors of NVG. ROC curve showed that the areas under curves of IL-6,IL-8,hs-CRP and the combined determination（IL-6+IL-8+hs-CRP） were 0.773,0.873,0.760 and 0.902,respectively. The efficiency of the combined determination was better than those of single determinations. Conclusions IL-6,IL-8 and hs-CRP are potential for the diagnosis of NVG,which play roles in the development of NVG.

Study on the main causes for false increasing of white blood cells and small round epithelial cells determined by UF-1000i urine sediment analyzer

ZHOU Xin, ZHANG Jue, CHEN Liying

2020, 35(5):  444-446.  DOI: 10.3969/j.issn.1673-8640.2020.05.010

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Objective To investigate the main causes for false increasing of white blood cells and small round epithelial cells determined by UF-1000i urine sediment analyzer. Methods Urine samples of outpatients were collected. UF-1000i urine sediment analyzer and microscopy were used,and the results of these 2 methods were compared. The main causes for false increasing of white blood cells and small round epithelial cells were evaluated. Results The false positive rate of white blood cells determined by UF-1000i urine sediment analyzer was 12.58%,mainly due to small round epithelial cells（69.6%）,bacteria（12.8%）,flat epithelial cells（8.0%） and fat droplets（6.4%）. The false positive rate of small round epithelial cells was 4.6%,mainly due to leukocytes（67.9%）,fat droplets（15.5%） and trichomoniasis（11.9%）. Conclusions When the urine pathological components increase,a variety of factors can affect UF-1000i urine sediment analyzer's determination results on white blood cells and small round epithelial cells,and false positivity appears. The application of UF-1000i urine sediment analyzer can only be a determination as a preliminary screening method,and microscopy is still be gold standard for urine sediment analysis.

Study on the main causes for false increasing of white blood cells and small round epithelial cells determined by UF-1000i urine sediment analyzer

ZHOU Xin, ZHANG Jue, CHEN Liying

2020, 35(5):  444-446.  DOI: 10.3969/j.issn.1673-8640.2020.05.010

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Objective To investigate the main causes for false increasing of white blood cells and small round epithelial cells determined by UF-1000i urine sediment analyzer. Methods Urine samples of outpatients were collected. UF-1000i urine sediment analyzer and microscopy were used,and the results of these 2 methods were compared. The main causes for false increasing of white blood cells and small round epithelial cells were evaluated. Results The false positive rate of white blood cells determined by UF-1000i urine sediment analyzer was 12.58%,mainly due to small round epithelial cells（69.6%）,bacteria（12.8%）,flat epithelial cells（8.0%） and fat droplets（6.4%）. The false positive rate of small round epithelial cells was 4.6%,mainly due to leukocytes（67.9%）,fat droplets（15.5%） and trichomoniasis（11.9%）. Conclusions When the urine pathological components increase,a variety of factors can affect UF-1000i urine sediment analyzer's determination results on white blood cells and small round epithelial cells,and false positivity appears. The application of UF-1000i urine sediment analyzer can only be a determination as a preliminary screening method,and microscopy is still be gold standard for urine sediment analysis.

Immune reconstitution after allogeneic hematopoietic stem cell transplantation and its correlation with fungal infection

PENG Xinguo, SUN Changhua, AN Jiajia, CUI Yanfang, DONG Yan, WANG Jian, GAO Meilan

2020, 35(5):  447-450.  DOI: 10.3969/j.issn.1673-8640.2020.05.011

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Objective To investigate the immune reconstitution of CD4⁺ T cells after allogeneic hematopoietic stem cell transplantation（allo-HSCT） and its correlation with invasive fungal infection（IFI） in patients with hematological malignancy. Methods A total of 47 patients with hematological malignancy and undergoing allo-HSCT in Binzhou Medical University Hospital from February 2010 to October 2014 were enrolled. A total of 40 healthy subjects were enrolled as control group. After transplantation for 1,2 and 3 months,the count of immune subpopulation and concentration of cytokines were determined by flow cytometry and enzyme-linked immunosorbent assay（ELISA）,respectively. The incidence of IFI after transplantation and its correlation with immune reconstitution of CD4⁺ T cells were observed. Results The counts of CD4⁺ T cells and immune subpopulation increased progressively after transplantation,and the immune subpopulation count after transplantation for 3 months was lower than that in control group. Compared with control group,the levels of interleukin（IL）-6 and IL-10 after transplantation increased,while that of transforming growth factor（TGF）-β decreased. The incidence rate of IFI was 19.15%（9/47）. Multivariate Logistic regression analysis showed that IFI might be related to helper T cell（Th）17 cell count instead of Th1,Th2 and regulatory T cell counts as well as IL-6,IL-10,TGF-β and interferon（IFN）-γ levels. Conclusions After allo-HSCT,the immune reconstitution of CD4⁺ T cells delays,and Th17 cell count decreases obviously,which may be related to the occurrence of IFI.

Analysis of co-infection of GBS and Mycoplasma in perinatal women

GUO Shuli, LUO Xiandao, JIA Haiying, YANG Liwei, HUANG Guohong, WANG Changmin

2020, 35(5):  451-454.  DOI: 10.3969/j.issn.1673-8640.2020.05.012

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Objective To analyze the characteristics of co-infection and drug susceptibility of group B Streptococcus（GBS） and Mycoplasma in perinatal women from 35 to 37 gestational weeks. Methods A total of 894 perinatal women from 35 to 37 gestational weeks in the People's Hospital of Xinjiang Uygur Autonomous Region from December 2017 to August 2018 were enrolled. Bacteria and fungi were determined by culture method. GBS was determined by broth enrichment culture automatic determination method,and Mycoplasma was determined by artificial culture method. Results In the 894 perinatal women,there were 65 cases of GBS infection and 41 cases of Ureaplasma urealyticum（Uu） infection,and there were 33 cases of co-infection. In bacterial infection（118 cases）,GBS was mainly,and in mixed infection（56 cases）,GBS+Uu was mainly. Totally,65 isolates of GBS were sensitive to linezolid,penicillin,vancomycin,meropenem,ampicillin and ceftriaxone,and the sensitive rates were 100%. The mediated isolates of levofloxacin,tetracycline and clindamycin were determined,and the drug resistance rate to clindamycin was the highest（86.15%）. Totally,41 isolates of Uu were sensitive to minocycline,doxycycline,roxithromycin,josamycin,and the sensitive rates were 100%. The mediated isolates of ciprofloxacin,sparfloxacin,ofloxacin,azithromycin,spectinomycin,levofloxacin and norfloxacin were determined,and the drug resistance rate to ciprofloxacin was the highest（92.68%）. Conclusions There are many infections of GBS and Uu in perinatal women in the People's Hospital of Xinjiang Uygur Autonomous Region,and the 2 infections are common. The drug resistance rate of GBS to clindamycin is high,and the drug resistance of Uu to ciprofloxacin is serious. Those should be taken seriously.

Establishment and preliminary clinical application of homogeneous method for the determination of remnant lipoprotein-cholesterol

WANG Rong, HUANG Keliang, ZHANG Jianhua, ZHANG Xin

2020, 35(5):  464-469.  DOI: 10.3969/j.issn.1673-8640.2020.05.016

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Objective To establish homogeneous method for determining remnant lipoprotein-cholesterol（RLP-C）,and to evaluate it primarily. Methods A homogeneous method for the determination of RLP-C was established based on the screening of non-lipoproteins with polyoxyethylene-polyethylene oxide butene（POE-POB） as screening agent. The self-established method was compared with the reference method [RLP-C reagent（homogeneous method） of Japan Concorde company]. Serum RLP-C and triglyceride（TG）,total cholesterol（TC）,apolipoprotein C2（apo C2） were determined in 116 patients with mixed hyperlipidemia and 130 healthy controls. Pearson correlation analysis was used to evaluate their correlations. Results The optimal buffer system was 3-morpholinopropanesulfonic acid（MOPS） buffer system. The optimal pH value was 6.8,the optimal concentration of POE-POB was 8 g/L,and the determination delay time was 5 min. Intra-self established method coefficient of value（CV） was 3.37%,and inter-CV was 4.91%. The reagent was stable for 1 month at 4 ℃. The results of reference method and self-established method were slightly different,but the correlation between the 2 methods was good（r=0.980）. Bland-Altman scatter plot showed that 4（4.08%） samples were not within ±1.96s. The concentrations of RLP-C,TG,apo C2 and TC in mixed hyperlipidemia group were higher than those in healthy control group（P<0.001）. RLP-C was positively correlated with TG（r=0.496,P<0.001）,but it was not correlated with apo C2 and TC（r=0.157 and -0.090,P>0.05）. Conclusions The self-established method is simple and rapid,which can be used in automatic biochemical analyzer and be used as clinical routine test.

Identification of vancomycin-resistant Enterococcus faecium by MALDI-TOF MS and epidemiological typing performance

WU Siying, WANG Xiaoqing, LIU Shanquan, XIAO Yuling, HE Chao, MA Ying, XIE Yi, KANG Mei

2020, 35(5):  470-475.  DOI: 10.3969/j.issn.1673-8640.2020.05.017

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Objective To identify vanA-positive vancomycin-resistant Enterococcus faecium（VREfm） and vancomycin-susceptible Enterococcus faecium（VSEfm） and determine the epidemiological correlation of VREfm by matrix-assisted laser desorption ionization-time of flight mass spectrometry（MALDI-TOF MS）. Methods A total of 139 Enterococcus faecium isolates were collected,and there were 66 VREfm and 73 VSEfm. Vancomycin resistance genes of 66 VREfm were confirmed by polymerase chain reaction（PCR）. Enterococcus faecium isolates were identified by MALDI-TOF MS. Models with ClinProTools 3.0 software were generated using the spectra of 45 VREfm and 50 VSEfm randomly selected. The remaining 21 VREfm and 23 VSEfm were validated with optimal model. Totally,21 VREfm randomly selected were analyzed by multilocus sequence typing（MLST） and subjected to hierarchical cluster analysis by MALDI-TOF MS. Results A total of 139 Enterococcus faeciumisolates were identified by MALDI-TOF MS,and the score range was 2.321-2.602. All the 66 VREfm carried vanA gene. External validation of the optimal model generated produced a sensitivity of 95.2% and a specificity of 95.7%. There were 7 sequence types（ST） of 21 VREfm,with most isolates identified as ST78. Hierarchical cluster analysis showed that 21 VREfm were classified into 4 clusters. Most of the ST78 isolates were classified into the same cluster,and highly-ST78-related ST were also classified into the ST78 cluster. Conclusions MALDI-TOF MS can rapidly differentiate vanA-positive VREfm from VSEfm with strict control of experimental conditions. Highly-related ST of VREfm are distinguishable.

Methylation level and mRNA expression of CpG island of HER2 gene in breast cancer and its adjacent tissues

JIN Shu, ZHU Qi, ZHU Yuan, YUAN Ya, CAI Xiaoyao, ZHANG Ji, YAN Peiyi

2020, 35(5):  481-486.  DOI: 10.3969/j.issn.1673-8640.2020.05.019

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Objective To investigate the methylation level and mRNA expression of CpG island of human epidermal growth factor receptor 2（HER2） gene in breast cancer and its adjacent tissues. Methods Totally,50 samples of breast cancer tissues and adjacent tissues confirmed by pathology were collected. The methylation rate of CpG island of HER2 promoter,the expression of HER2 mRNA and the expression of HER2 protein were determined by pyrosequencing,real-time polymerase chain reaction（PCR） and immunohistochemistry,respectively. Results The relative expression of HER2 mRNA in HER2 positive breast cancer tissues was higher than that in HER2 negative breast cancer tissues（P<0.05）,and there was no statistical significance in the methylation rate of CpG island of HER2 promoter（P>0.05）. The methylation rate of HER2 promoter CpG island in HER2 negative breast cancer tissues was higher than that in adjacent tissues（P<0.05）,while the expression of HER2 mRNA was lower than that in adjacent tissues（P<0.05）. There was no statistical significance in the methylation rate of CpG island of HER2 promoter and the expression of HER2 mRNA between HER2 positive breast cancer and adjacent tissues and between the 2 groups of adjacent tissues（P>0.05）. Conclusions Hypomethylation of CpG island of HER2 promoter may be one of the mechanisms of high expression of HER2 mRNA in breast cancer adjacent tissues of HER2 negative group.

Investigation on the cut-off values of tandem mass spectrometry for neonatal inherited metabolic diseases in China

HE Falin, LIN Bo, WANG Wei, ZHONG Kun, YUAN Shuai, DU Yuxuan, WANG Zhiguo

2020, 35(5):  487-491.  DOI: 10.3969/j.issn.1673-8640.2020.05.020

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Objective To investigate the cut-off values of 16 indicators for screening neonatal inherited metabolic diseases by tandem mass spectrometry in China,and to provide a reference on setting cut-off values for screening neonatal inherited metabolic diseases. Methods The data related to the cut-off values of 4 amino acids and 12 acylcarnitines from 141 neonatal screening laboratories,including laboratory coding,cut-off value,source of cut-off value,methodological principle,instrument,reagent,calibrator,modifying or not,were collected. The effects of different methods and sources of cut-off values on the 16 cut-off values were analyzed. Results A total of 114 laboratories submitted their data. Totally,70.18% laboratories' cut-off values were from reagent manufacturer's instructions,about 16% laboratories' cut-off values were determined by own laboratory,and about 14% laboratories' cut-off values were from literature reports. The proportion of laboratories with modified cut-off values was 49.12%-53.51%. The cut-off values had statistical significance between derivative method and non-derivative method（P<0.05）. The coefficient of variation of derivative method was distributed in 25.26%-172.05%,and the dispersion degree of unmodified tangent value was greater than that of modified tangent value. The coefficient of variation of non-derivative method was distributed in 18.15%-200.92%,and the unmodified tangent values were consistent with the dispersion degree of modified tangent values except low citrulline values. Conclusions There are differences in the cut-off value of tandem mass spectrometry in screening neonatal inherited metabolic diseases in China,and the cut-off value should be evaluated and validated regularly.

Primary experience of establishment and management of medical laboratory in cabin hospital under the background of COVID-19

YU Siyuan, WU Wenjuan

2020, 35(5):  492-496.  DOI: 10.3969/j.issn.1673-8640.2020.05.021

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Under the background of corona virus disease 2019（COVID-19） epidemic situation,a large number of patients can not be treated in designated hospitals due to local hospitals' limited beds. The isolation and treatment of diagnosed mild and general COVID-19 patients in cabin hospital and cutting off the source of infection are the keys to controlling the epidemic. The establishment and management of medical laboratory in cabin hospital provides scientific and objective laboratory data for clinic,which is an indispensable part of the prevention and control measures of COVID-19. In this review,the site selection,personnel and training,infection prevention,quality control and process optimization of medical laboratory are summarized,which provides a reference for other cabin hospitals and clearing hospitals.

Evaluation of repeat-sampling quality control strategy based on 1:2s rule

ZHANG Yu, PENG Aihong, YANG Lihua, YU Qihua

2020, 35(5):  497-500.  DOI: 10.3969/j.issn.1673-8640.2020.05.022

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The application of repeat-sampling quality control strategy based on 1:2s rule has been discussed,and its performance has been evaluated. Office software is used to assist laboratories in actual operation,in order to recommend a simple,efficient and manual application of internal quality control strategy.

Research progress on neutrophil to lymphocyte ratio in eye diseases

MA Yingbo, CAO Wenjun

2020, 35(5):  501-505.  DOI: 10.3969/j.issn.1673-8640.2020.05.023

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Peripheral blood neutrophil to lymphocyte ratio（NLR） is an immune inflammation indicator. Abnormal elevation of NLR indicates poor prognosis of malignant tumors,autoimmune diseases and cardiovascular diseases. Recently,it has been suggested that low-level immune inflammation is probably involved in eye diseases,such as glaucoma,diabetic retinopathy（DR）,macular degeneration and dry eye disease（DED）. Many scholars have explored the relationship between NLR and various eye diseases. In this review,the main clinical application of NLR and its research progress in some eye diseases are summarized.