Laboratory Medicine

Application of serum bile acid spectrum in the differential diagnosis of pneumonia and lung cancer

XU Runhao, ZOU Chen, ZHANG Jie, LI Min, ZHANG Shulin

2021, 36(1): 1-7. DOI: 10.3969/j.issn.1673-8640.2021.01.001

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Objective To investigate the changes of serum bile acid spectrum in patients with lung cancer and its value in the differential diagnosis of pneumonia and lung cancer. Methods Serum bile acid spectrum of 80 patients with pneumonia（pneumonia group）,108 patients with lung cancer（lung cancer group） and 106 healthy subjects（healthy control group） were determined by liquid chromatography-tandem mass spectrometry（LC-MS/MS）,including 5 free bile acids [cholic acid（CA）,chenodeoxycholic acid（CDCA）,deoxycholic acid（DCA）,lithocholic acid（LCA）,ursodeoxycholic acid（UDCA）] and 10 conjugated bile acids [glycocholic acid（GCA）,glycochenodeoxycholic acid（GCDCA）,glycodesoxycholic acid（GDCA）,glycolithocholic acid（GLCA）,glycoursodeoxycholic acid（GUDCA）,taurocholic acid（TCA）,taurochenodeoxycholic acid（TCDCA）,taurodeoxycholic acid（TDCA）,taurolithocholic acid（TLCA）and tauroursodeoxycholic acid（TUDCA）]. Serum total bile acid（TBA） and tumor markers,such as carbohydrate antigen（CA） 125,CA19-9,carcinoembryonic antigen（CEA）,cytokeratin 19 fragment（CYFRA 21-1） and neuron specific enolase（NSE）,were also determined. Logistic regression was used to screen items,and the diagnosis model was established. Receiver operating characteristic（ROC） curve was used to analyze the performance of these items and diagnosis model in the differential diagnostic of pneumonia and lung cancer. Results Compared with healthy control group,the levels of DCA,LCA,UDCA,GDCA,GLCA,TDCA,and TLCA decreased in pneumonia group（P<0.01）,while GCDCA and TCDCA increased（P<0.01）; CA,CDCA,GCDCA,and TCDCA increased in lung cancer group（P<0.01）,while TDCA and TLCA decreased（P<0.01）. There was no statistical significance in the level of serum TBA among the 3 groups（P>0.05）. The levels of CA,CDCA,DCA,LCA,UDCA,GDCA,GLCA,CEA,and CYFRA21-1 in lung cancer group were significantly higher than those in pneumonia group（P<0.01）. ROC curve analysis showed that DCA had the best performance in the differential diagnosis of pneumonia and lung cancer among all the single items [the area under curve（AUC） of 0.781,the sensitivity of 73.2%,the specificity of 77.5% and the Youden index of 0.51]. The diagnosis model had good performance with the AUC of 0.873,the optimal cut-off value of 0.52,the sensitivity of 89.3% and the specificity of 75.4%. Conclusion The changes of bile acid spectrum are closely associated with the occurrence of lung diseases. Bile acid spectrum is potential as diagnostic markers for lung cancer.

Establishment of reference interval for the detection of serum protein electrophoresis by V8 Nexus high speed automated capillary electrophoresis apparatus

AN Chongwen, LI Haixia, JIAO Lili

2021, 36(1): 8-12. DOI: 10.3969/j.issn.1673-8640.2021.01.002

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Objective To preliminarily establish a reference interval for serum protein electrophoresis（SPE） by V8 Nexus high speed automated capillary electrophoresis（CE） apparatus. Methods A total of 649 healthy subjects were enrolled in this study and classified into 5 groups according to ages,including 18-29-year-old,30-39-year-old,40-49-year-old,50-59-year-old and ≥60-year-old groups. Serum samples were detected by the CE apparatus. The reference interval of CE for SPE detection was evaluated according to the document method of the Health Industry Standard WS/T 402—2012 of the People's Republic of China. Results All data from males and females obeyed normal distribution or approximately normal distribution（P>0.05）. There were statistical differences in the levels of albumin,α₂ globulin,β₁ globulin and γ globulin between males and females（P<0.05）,while there was no statistical significance in the results of α₁globulin,β₂ globulin or β globulin between males and females（P>0.05）. In males,there were statistical differences in the levels of albumin,α₂ globulin and β₁ globulin between 18-59-year-old groups and ≥60-year-old group（P<0.05）,but there was no statistical significance in these parameters between 18-59-year-old groups（P>0.05）;neither were α₁ globulin,β₂ globulin,γ globulin or β globulin between any groups（P>0.05）. In females,there were statistical differences in the levels of albumin,β₁ globulin,β₂ globulin and β globulin between 18-59-year-old groups and ≥60-year-old group（P<0.05）,but there was no statistical significance in these parameters between 18-59-year-old groups（P>0.05）;neither were α₁ globulin,α₂ globulin or γ globulin between any groups（P>0.05）. According to the results of LSD test,SNK test and Z test,the age groups could be combined into one group to establish the reference interval of SPE. Conclusion The reference interval of SPE has been preliminarily established by V8 Nexus high speed automated CE apparatus. Clinical laboratories should establish a reference interval for different sexes or ages according to actual situation.

Clinical role of combined application of different eGFR equations and ACR in high-risk renal dysfunction patients

XU Wenjun, LI Jiayong, LI Huidan

2021, 36(1): 13-19. DOI: 10.3969/j.issn.1673-8640.2021.01.003

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Objective To investigate the combined application of different estimating glomerular filtration rate（eGFR） equations and urinary albumin to creatinine ratio（ACR） in high-risk renal injury or early renal dysfunction patients. Methods Clinical data of 850 patients with chronic kidney disease（CKD） were collected,including sex,age,height,weight,serum creatinine（SCr）,cystatin C（Cys C）,ACR,serum urea,serum uric acid（UA）,glomerular filtration rate measured by ^99mtechnetium-diethyenetriamine pentaacetic acid（^99mTC-DTPA） renal dynamic imaging measuring glomerular filtration rate（mGFR） and basic medical history. Totally,3 SCr-based equations（CKD-EPI 2009Scr,simplified MDRD and improved MDRD）,6 equations based on Cys C（CKD-EPI 2012Cys C,Grubb,Arnal-Dade,Rule,Macisaac and Tan） and 1 equation based on Cys C combined with SCr（CKD-EPI 2012SCr-Cys C）were applied respectively to calculate eGFR of all the subjects. The correlations between different eGFRs and between different eGFRs and mGFR were analyzed by Spearman correlation analysis. Bland-Altman analysis was used to evaluate the consistency between eGFR and mGFR. Results In male and female groups,the level of eGFR based on SCr（eGFR-SCr） was higher than that of mGFR（P<0.01）,and the level of eGFR based on Cys C（eGFR-Cys C） was lower than that of mGFR（P<0.01）. There were positive correlations among eGFR-SCr,among eGFR-Cys C,between eGFR and mGFR（P<0.01）. The average deviation between eGFR-SCr and mGFR was –4.2-20.8 mL/（min•1.73 m²）,which overestimated patient's GFR. Between eGFR-Cys C and mGFR,the average deviation was 3.7-16.9 mL/（min•1.73 m²）,which underestimated the GFR. Between eGFR_{CKD-EPI 2012SCr-Cys C} and mGFR,the average deviation was 9.6 mL/（min•1.73 m²）,which was also lower than the actual GFR. For patients with G2 stage which were missed by eGFR_{CKD-EPI 2009SCr},79.2% and 55.8% was detected by eGFR_Macisaac and ACR,respectively,and the combination of eGFR_Macisaac and ACR was 87.7%. Furthermore,for patients with G3 to G5 stage missed by eGFR_{CKD-EPI 2009SCr},43.3% and 61.1% were detected with eGFR_Macisaac and ACR,respectively,and 73.3% were detected by the combination of eGFR_Macisaac and ACR. Compared with eGFR_{CKD-EPI 2009SCr} alone,the detection rate of G2 stage was increased to 94.4%,and the detection of G3-G5 stage was increased to 91.7% after combining eGFR_Macisaac and ACR. Conclusion The combined application of eGFR_{CKD-EPI 2009SCr},eGFRMacisaac and ACR can significantly improve the detection rate of high-risk renal impairment and early renal dysfunction.

Roles of SCC-Ag,CA19-9 and VEGF for chemotherapy efficacy in advanced esophageal squamous cell carcinoma

YANG Yong

2021, 36(1): 20-24. DOI: 10.3969/j.issn.1673-8640.2021.01.004

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Objective To investigate the roles of squamous cell carcinoma antigen（SCC-Ag）,carbohydrate antigen 19-9（CA19-9） and vascular endothelial growth factor（VEGF） for chemotherapy efficacy in advanced esophageal squamous cell carcinoma. Methods Totally,112 patients with advanced esophageal squamous cell carcinoma were enrolled. All the patients were treated with docetaxel + nedaplatin. Serum levels of SCC-Ag,CA19-9 and VEGF before and after chemotherapy were determined by enzyme-linked immunosorbent assay,and the chemotherapy efficacy of SCC-Ag,CA19-9 and VEGF was evaluated by receiver operating characteristic（ROC） curve. Results The effective rate of chemotherapy in 112 patients with esophageal squamous cell carcinoma was 53.57%（60/112）. There was no statistical significance in serum levels of SCC-Ag,CA19-9 and VEGF between effective chemotherapy group and ineffective chemotherapy group（P>0.05）. Serum levels of SCC-Ag,CA19-9 and VEGF in effective chemotherapy group after chemotherapy were lower than those before chemotherapy（P<0.001）,and serum levels of SCC-Ag,CA19-9 and VEGF in ineffective chemotherapy group after chemotherapy were higher than those before chemotherapy（P<0.001）. The suppression ratios of SCC-Ag,CA19-9 and VEGF in effective chemotherapy group were higher than those in ineffective chemotherapy group（P<0.001）. ROC curve analysis results showed that the area under curve（AUC） of the suppression ratios of SCC-Ag,CA19-9 and VEGF in evaluating the chemotherapy efficacy of esophageal squamous cell carcinoma were 0.741,0.712 and 0.783,the optimal cut-off values were 20%,26% and 27%,the sensitivities were 76.67%,80.00% and 83.33%,the specificities were 76.92%,73.08% and 80.77%,respectively. The AUC of the combination of the 3 factors（SCC-Ag suppression ratio >20%,CA19-9 suppression ratio >26%,VEGF suppression ratio >27%） in evaluating the chemotherapy efficacy of esophageal squamous cell carcinoma was 0.926,the sensitivity was 96.67%,and the specificity was 96.15%. Conclusions SCC-Ag,CA19-9 and VEGF can be used to evaluate the chemotherapy efficacy of advanced esophageal squamous cell carcinoma.

Relationship between complement 3,interleukin 35 and the severity and prognosis of sepsis

HU Senan, AI Honghong, YE Xuelian, CHENG Jiangyan

2021, 36(1): 25-29. DOI: 10.3969/j.issn.1673-8640.2021.01.005

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Objective To analyze the relationship between complement 3（C3）,interleukin 35（IL-35）and the severity and prognosis of sepsis. Methods A total of 80 patients with sepsis were enrolled in this study,including 45 cases of general sepsis（general sepsis group）and 35 cases of severe sepsis（severe sepsis group）. In the severe sepsis group,there were 12 cases of single organ dysfunction（single organ dysfunction subgroup）,16 cases of multiple organ dysfunction syndrome（MODS subgroup）and 7 cases of death（death subgroup）. Clinical data of all patients were collected. Blood routine test [white blood cell（WBC） count,platelet（PLT） count],IgG,IgA,IgM,C3,C4,prothrombin time（PT）,international standardized ratio（INR）,activated partial thromboplastin time（APTT）,fibrinogen（Fib）,alanine aminotransferase（ALT）,total bilirubin（TB）,creatinine（Cr）,creatine kinase isoenzyme MB（CK-MB） and IL-35 were determined within 24 h after patients were transferred to intensive care unit（ICU）. Meanwhie, patients were evaluated by the acute physiology and chronic health evaluation Ⅱ（APACHEⅡ）,and were divided into different groups（APACHEⅡscore≤25 and APACHEⅡ score>25） according to the APACHEⅡ score. Results There were significant differences in the C3 and IL-35 between general sepsis group and severe sepsis group（P<0.05）. There were also significant differences in the C3 and IL-35 between subgroups and general sepsis group（P=0.001,P<0.001）. With the progression of the disease,C3 decreased and IL-35 increased. C3 was positively correlated with PLT and Fib（r=0.32 and 0.52,P<0.001）,and negatively correlated with PT,INR,APTT,ALT,TB,Cr and CK-MB（r=-0.34,-0.38,-0.33,-0.31,-0.27,-0.25 and -0.26,P<0.001）. IL-35 was negatively correlated with PLT and Fib（r=-0.33 and -0.59,P<0.001）,and positively correlated with PT,INR,APTT,ALT,TB,Cr and CK-MB（r=0.41,0.32,0.25,0.42、0.34,0.29 and 0.26,P<0.001）. There were significant differences in the C3 and IL-35 levels between patients with APACHEⅡscore≤25 and those with APACHEⅡ score>25（P<0.05,P<0.001）. C3 was negatively correlated with prognosis（death）（r=-0.63,P<0.05）,and IL-35 was positively correlated with prognosis（death）（r=0.54,P<0.001）. Conclusion C3 and IL-35 are involved in the coagulation disorders and organ damage in sepsis patients. They may be used to evaluate the condition and prognosis of sepsis.

Roles of ET-1,EMP and hs-cTnT in ischemic heart disease

ZHANG Shenwei, GUO Jing, ZHANG Zheng, CHEN Jianghong, ZHAO Kui

2021, 36(1): 30-33. DOI: 10.3969/j.issn.1673-8640.2021.01.006

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Objective To investigate the roles of endothelin-1（ET-1）,endothelial microparticle（EMP） and high-sensitivity cardiac troponin T（hs-cTnT） in patients with ischemic heart disease. Methods Serum levels of ET-1,EMP and hs-cTnT were determined in 98 patients with ischemic heart disease（ischemic heart disease group）and 100 healthy subjects（healthy control group）. According to the New York Heart Association（NYHA） cardiac function classification,there were 27 patients with NYHA class Ⅰ,34 patients with class Ⅱ,25 patients with class Ⅲ and 12 patients with class Ⅳ. According to Gensini score,98 patients were classified into mild stenosis（32 cases）,moderate stenosis（45 cases） and severe stenosis（21 cases）. Results The levels of ET-1,EMP and hs-cTnT in ischemic heart disease group were significantly higher than those in healthy control group（P<0.05）. The levels of ET-1,EMP and hs-cTnT in patients with NYHA class Ⅳ and Ⅲ were significantly higher than those in patients with NYHA class Ⅰ and Ⅱ（P<0.05）. Moreover,patients with NYHA class Ⅳ had higher levels of ET-1,EMP and hs-cTnT than those with NYHA class Ⅲ（P<0.05）. The levels of ET-1,EMP and hs-cTnT in patients with mild,moderate and severe stenosis increased in turn（P<0.05）. Conclusion The levels of ET-1,EMP and hs-cTnT are correlated with the grade of cardiac function and the degree of coronary artery stenosis in patients with ischemic heart disease.

Diagnostic value of miR-124a and miR-449a in plasma of non-small cell lung cancer

PENG Jiaowu, SUN Chengmou

2021, 36(1): 34-38. DOI: 10.3969/j.issn.1673-8640.2021.01.007

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Objective To explore the diagnostic value of microRNA-124a（miR-124a）and microRNA-449a（miR-449a） in non-small cell lung cancer（NSCLC）. Methods Ninety patients with NSCLC who underwent surgical resection and were followed up for 36 months were assigned to NSCLC group and subdivided into relapsed group（56 cases）and non-relapsed group（34 cases）. In addition,60 patients with benign lung nodules（benign nodules group） and 80 healthy persons（normal control group） were enrolled in this study. General information of all the subjects was collected. Serum neuron specific enolase（NSE）,miR-124a and miR-449a levels were detected. Spearman correlation analysis was applied to evaluate the relevance of indicators. The receiver operating characteristic（ROC） curve was used to evaluate the effectiveness of the indicators in the diagnosis of NSCLC. Results Compared with those in the normal control group and benign nodule group, serum NSE was significantly increased in the NSCLC group （P<0.05）,but the serum levels of miR-124a and miR-449a were significantly decreased（P<0.05）. There was no statistically significant difference in serum levels of NSE,miR-124a or miR-449a between the normal control group and benign nodule group（P>0.05）. Compared with those in the non-relapsed group,serum NSE of the relapsed group increased（P<0.05）,while the levels of miR-124a and miR-449a decreased（P<0.05）. There was no significant difference in age,gender,body mass index（BMI）,smoking status,tumor diameter,pathological type and TNM stage between the two groups（P>0.05）. ROC curve analysis results showed that the area under curve（AUC）of NSE,miR-124a and miR-449a for single diagnosis of NSCLC were 0.660,0.703 and 0.759,respectively. The combined detection of NSE+miR-124a+miR-449a had the highest AUC（0.895） and sensitivity（96.25%） for the diagnosis of NSCLC. The combined detection of miR-124a+miR-449a had the highest specificity（90.31%） for the diagnosis of NSCLC. Conclusion miR-124a and miR-449a may be used as auxiliary diagnostic indicators for NSCLC.

Diagnostic value of SARS-CoV-2 antibody to COVID-19：a meta-analysis

LI Shichao, SHEN Liang, WANG Chunhua, TONG Yanfei, ZHANG Ji

2021, 36(1): 39-43. DOI: 10.3969/j.issn.1673-8640.2021.01.008

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Objective To investigate the value of severe acute respiratory syndrome coronavirus 2（SARS-CoV-2） antibody in the diagnosis of corona virus disease 2019（COVID-19） by meta-analysis. Methods Relevant articles were collected from PubMed,Embase,the Cochrane Library,CNKI,Wanfang Data and VIP databases. The pooled sensitivity,specificity,diagnostic odds ratio（DOR） were calculated,and the heterogeneity was investigated. The area under the summary receiver operating characteristic（SROC） curve was used to evaluate the diagnostic efficacy of SARS-CoV-2 antibody for COVID-19. Results Totally,11 studies including 1 117 COVID-19 patients and 713 healthy controls were collected in this meta-analysis. The pooled of sensitivity,specificity,DOR and area under curve（AUC） of SARS-CoV-2 antibody IgM were 0.82,0.97,139.95 and 0.98,and those of SARS-CoV-2 antibody IgG were 0.83,0.98,213.02 and 0.98,respectively. Chemiluminescence had higher sensitivity than colloidal gold method. The heterogeneity was evident in these studies,but there was no threshold effect. Conclusion SARS-CoV-2 IgM and IgG antibodies have high values in the auxiliary diagnosis and differential diagnosis of COVID-19.

Analysis on the absolute value of lymphocyte in COVID-19 patients before and after recovery in Shanghai

FAN Jian, GUO Ying, QI Weiqiang, HAO Menglu, ZHANG Tengfei, ZHU Zhaoqin

2021, 36(1): 44-47. DOI: 10.3969/j.issn.1673-8640.2021.01.009

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Objective To evaluate the role of absolute lymphocyte count（LYMPH#）in predicting prognosis of corona virus disease 2019（COVID-19）patients. Methods A total of 123 COVID-19 patients were enrolled in this retrospective study. There were 112 mild cases and 11 severe cases. The LYMPH# in peripheral blood were collected at the initial hospitalization,before discharge,and at the initial follow-up. Clinical data of all the patients were also collected. Results Severe patients were older than mild patients. At the initial hospitalization,the LYMPH# of 50.89%（57/112）of mild group was lower than normal value,and the LYMPH# of 72.73%（8/11）of severe group was lower than referencevalue. There was no significant difference in the LYMPH# between mild group and severe group at the initial hospitalization（P=0.789）. There was significant difference in the recovery LYMPH# [（initial follow-up results-first admission results）/initial follow-up results] between the two groups（P=0.014）. There was the difference of LYMPH# between before discharge and the first follow-up（P=0.038）. Conclusion The cellular immune function of COVID-19 patients is generally low,and needs continuous monitoring. LYMPH# can be used as an indicator to monitor the immune function and predict the prognosis of COVID-19 patients.

Correlation between some laboratory indexes and severity of COVID-19 in 144 patients

LIAO Shengjun, XU Xianqun, ZHAO Jin, LI Siwei, QIU Xueping, CHEN Liangjun, LI Yirong

2021, 36(1): 48-52. DOI: 10.3969/j.issn.1673-8640.2021.01.010

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Objective To study the correlation between some laboratory indexes and severity of corona virus disease 2019（COVID-19）. Methods The results of laboratory examination of 144 patients with COVID-19 confirmed in Zhongnan Hospital of Wuhan University from January 20,2020 to March 10,2020 were collected and analyzed retrospectively. The blood routine examination,D-dimer（DD）,C-reactive protein（CRP）,procalcitonin（PCT）,serum amyloid A protein（SAA）,cytokines and lymphocyte subgroups were compared between common and severe COVID-19 patients. Logistic regression was used to analyze the relationship between laboratory indexes and COVID-19 severity. Results Severe patients had higher levels of white blood cell（WBC） count,neutrophil percentage （NEUT%）,neutrophil/lymphocyte ratio（NLR）,SAA,CRP,PCT,DD,interleukin（IL）-6 and IL-10 than commonpatients（P<0.05）. Lymphocyte percentage（LYMPH%） was significantly lower in severe group than that in common group（P<0.05）. There was no significant difference in mean corpuscular volume（MCV）,red blood cell volume distribution width（RDW）,hemoglobin（Hb）,platelet（PLT） count,tumor necrosis factor alpha（TNF-α）,IL-2 or IL-4 between the two groups（P>0.05）. The total lymphocyte number was significantly lower in severe group than that in common group（P<0.05）,but no significant difference was found in the lymphocyte constituent ratio between the two groups（P>0.05）. Logistic regression analysis showed that WBC count,NEUT%,NLR,SAA,DD,CRP,IL-6 and IL-10 could be used as laboratory indexes to reflect the severity of COVID-19（P<0.05）. Conclusion WBC count,NEUT%,NLR,SAA,DD,CRP,IL-6 and IL-10 in severe COVID-19 patients have obvious changes as compared with those in common patients. The changes of these laboratory indexes are helpful for the early diagnosis of COVID-19.

Early diagnosis value of NLR,PCT and CRP combined determination for COVID-19

ZHU Chuanxin, ZHOU Yuping, ZHOU Qin, ZHOU Guifeng

2021, 36(1): 53-56. DOI: 10.3969/j.issn.1673-8640.2021.01.011

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Objective To investigate the value of neutrophil/lymphocyte ratio（NLR）,procalcitonin（PCT） and C-reactive protein（CRP） determinations in the early diagnosis of corona virus disease 2019（COVID-19）. Methods A total of 230 suspected COVID-19 patients were enrolled and classified into COVID-19 group（80 cases）,cold group（100 cases） and bacterial group（50 cases）according to thorax computed tomography（CT）,severe acute respiratory syndrome coronavirus 2（SARS-CoV-2） nucleic acid,viral serological antibody and blood culture results. The levels of white blood cell（WBC） count,the absolute value of neutrophil（NEUT#）,the absolute value of lymphocyte（LYMPH#）,the percentage of lymphocyte（LYMPH%）,the absolute value of easinosphils（EO#）,NLR,PCT and CRP among the 3 groups were compared. Results Compared with cold group,COVID-19 group had higher levels of NLR and CRP,and the levels of LYMPH%,LYMPH# and EO# were lower（P<0.05）. Compared with bacterial group,COVID-19 group had lower levels of LYMPH#,EO#,NEUT#,PCT and WBC count（P<0.05）. Compared with cold group,COVID-19 group had higher proportions of WBC count<4.00,LYMPH#<0.80,LYMPH%<20%,EO#<0.05,NLR>3.0 and CRP≥10.00（P<0.05）. Compared with bacterial group,COVID-19 group had higher proportions of WBC count<4.00,NEUT#<2.00,LYMPH#<0.80 and EO#<0.05（P<0.05）and lower proportions of WBC count>10.00,NEUT#>7.00,EO#>0.50,PCT≥0.50（P<0.05）. Conclusions Decreased LYMPH#,normal PCT and increased NLR and CRP are early markers of COVID-19 patients. The combined determination can be used as a supplement to SARS-CoV-2 nucleic acid determination.

Prediction of severe corona virus disease 2019 by coagulation indicators

WANG Xiaowu, GAO Yong, WU Dong

2021, 36(1): 57-59. DOI: 10.3969/j.issn.1673-8640.2021.01.012

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Objective To investigate the roles of prothrombin time（PT）,activated partial thromboplastin time（APTT）,fibrinogen（Fib）,thrombin time（TT）and D-dimer（DD） in the prediction and diagnosis of severe corona virus disease 2019（COVID-19）. Methods A total of 120 patients with COVID-19 were recruited from Fuyang Second People's Hospital from January 2020 to February 2020. There were 17 cases of severe group and 103 cases of mild group. The differences of PT,APTT,Fib,TT and DD were compared between the 2 groups,and a predictive model was established by Logistic regression analysis and evaluated by receiver operating characteristic（ROC） curve analysis. Results Compared with mild group,severe group had higher TT,DD and Fib（P<0.01,P<0.05）. DD and TT were independent risk factors for severe COVID-19. The predictive model Logit（P）=-12.460+1.760DD+0.601TT was established,and the goodness of fit test results of the predictive model showed P>0.05. The areas under curves（AUC）of DD,TT and combined determinations were 0.854,0.817 and 0.875,respectively. The sensitivities were 76.47%,70.59% and 70.59%,and the specificities were 83.50%,86.41% and 98.06%,respectively. DD predicted that the cut-off value of severe COVID-19 was 0.48 μg/L,and the cut-off value of TT was 16 s. Conclusions DD>0.48 μg/L and TT>16 s can effectively predict and diagnose the occurrence of severe COVID-19. The performance of combined determination is better.

Construction of colon cancer prognostic model based on TCGA SpliceSeq alternative splicing events

LEI Ming, GUO Mengyue, WANG Ruoying, NI Xiaomei, SHI Qiong

2021, 36(1): 60-68. DOI: 10.3969/j.issn.1673-8640.2021.01.013

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Objective To establish a genome-wide alternative splicing event risk model to predict the prognosis of colon adenocarcinoma（COAD）. Methods COAD RNA-Seq data and clinical information were downloaded from the Cancer Genome Atlas（TCGA）,and alternative splicing events were obtained from TCGA SpliceSeq database. Splice factors were downloaded from the SpliceAid 2 database. Univariate Cox regression analysis was employed to determine prognostic related alternative splicing events（PASE）. Lasso regression was used to screenvariables. Multivariate Cox regression was performed to calculate risk score and constructmodels. Cytoscape Reactome FI plug-in was used to build an interactive network and find core nodes. GO and KEGG were implemented for gene function annotation and pathway analysis. Kaplan-Meier and receiver operating characteristic （ROC）curve were used for the evaluation of PASE risk model. PASE of SF and other genes were constructed to form prognosis-related interactive network. Results A total of 35 391 mRNA alternative splicing events occurred in 9 085 genes from 398 COAD patients,and 2 015 surviving PASEs occurred in 1 811 genes. In the risk model made by 8 PASE,patients were divided into high-risk group and low-risk group according to the optimal cut-off value of 0.919,and there were statistical difference between groups （P<0.001）. The area under the ROC curve of the risk model was 0.860 (one year survival rate). In univariate Cox regression analysis,tumor invasion,lymph node metastasis,distant metastasis,clinical stages and risk model were significantly negatively correlated with overall survival time（P<0.001）. After adjusting for multiple factors,the risk model still showed a significant negative correlation with the overall survival time of patients（P<0.001）. In the risk model,there was no correlation between 8 PASE and mRNA expressions of their corresponding genes（P>0.05）. Conclusion The effect of PASE on COAD prognosis is investigated by TCGA-COAD genome-wide analysis and a risk model has been constructed that can be used to predict clinical prognosis.

Clinical significance of inflammatory cell types and inflammatory cytokines in sputum induced by bronchial asthma in children

GAO Weixia, QIN Xiaowan, LIU Yang, ZHANG Jing, DING Xianchun

2021, 36(1): 69-74. DOI: 10.3969/j.issn.1673-8640.2021.01.014

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Objective To explore the clinical value of inflammatory cell types and inflammatory cytokines in induced sputum in children with bronchial asthma. Methods Fifty-four children with bronchial asthma were selected as the research objects（study group）,and subdivided into mild asthma group,moderate asthma group,and severe asthma group. Forty-two sex- and age-matched children with bronchopneumonia（without asthma）were selected into control group. The inflammatory cell types,T cell immunoglobulin and mucin-3 （Tim-3） mRNA,and inflammation-related cytokines [interleukin（IL）-2,IL-4,IL-10,IL-13,interferon gamma（IFN-γ）] in induced sputum were detected before and after treatment. Results Before treatment,in the induced sputum,eosinophilratio,lymphocyte ratio,Tim-3 mRNA,IL-4,IL-13,IFN-γ and IFN-γ/IL-10 in the study group were significantly higher than those in the control group（P<0.05）,while the levels of IL-2,IL-10 and Th1/Th2 in the study group were significantly lower than those in the control group（P<0.05）. Children with moderate and severe asthma had higher levels of the percentage of eosinophil,the percentage of neutrophil,the percentage of lymphocyte,Tim-3 mRNA,IL-4,IL-13,IFN-γ and IFN-γ/IL-10 than children with mild asthma and children with bronchopneumonia（P<0.05）,while IL-2,IL-10 and Th1/Th2 in the children with moderate and severe asthma were significantly lower than those in the children with mild asthma and children with bronchopneumonia（P<0.05）. Logistic regression analysis showed that the proportion of eosinophils,Tim-3 mRNA,IFN-γ,IL-10,IFN-γ/IL-10 and Th1/Th2 were independent influencing factors of bronchial asthma in children. Receiver operating characteristic （ROC） curve analysis showed that the percentage of eosinophils,Tim-3 mRNA,IFN-γ,IFN-γ/IL-10 and Th1/Th2 had a certain differential effect on bronchial asthma and bronchial pneumonia in children. Conclusion Abnormal expression of eosinophils,Tim-3 mRNA,IFN-γ,IFN-γ/IL-10 and Th1/Th2 in induced sputum of children with bronchial asthma can help to monitor the disease condition,evaluate the efficacy and discriminatebronchopneumonia in children without asthma.

Establishment and evaluation of fast predictive algorithm for urinary tract infection pathogen in patients with nephrolithiasis

XIAO Nan, ZHOU Jia, DUAN Ning, ZHU Tianyang, WANG Lijun

2021, 36(1): 75-79. DOI: 10.3969/j.issn.1673-8640.2021.01.015

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Objective To establish an algorithm that can predict potential pathogens causing urinary tract infection（UTI）based on routine urinalysis in nephrolithiasis patients. Methods A total of 414 clean-catch urine specimens of nephrolithiasis patients were collected. The comparison between flow cytometry count,nitrite,leukocyte esterase and urine culture was undertaken to assess the optimal cut-off value for predictive algorithm. Results The cut-off value of flow cytometry bacterial count was 231.8/μL（sensitivity 73.97%,specificity 91.14%）. Furthermore,the samples with nitrite positive（OR=7.789,95%CI: 3.304-18.358,P<0.000 1） and/or leukocyte esterase 4+（OR=2.738,95%CI: 1.358-5.519,P=0.004 9） were more likely to contain Gram negative bacteria. Conclusion A fast predictive algorithmusing the combination of urine dipstick and flow cytometry can predict the infection of Gram-positive bacteria and Gram-negative bacteria in nephrolithiasis patientsand has certain clinical value.

Serum high mobility group box-1 expression in patients with sepsis caused by bacteria and its clinical significant

FU Jing, TU Xing

2021, 36(1): 80-83. DOI: 10.3969/j.issn.1673-8640.2021.01.016

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Objective To observe the predictive role of high mobility group box 1 protein（HMGB1）in the survival status of patients with bacterialsepsis. Methods A total of 110 patients with sepsis caused by bacteria confirmed from June 2015 to June 2017were recruited in this study. They were divided into survival group（n=84）and death group（n=26）according to survival status. The plasma level of HMGB1 was measured using ELISA method. The distribution and composition of pathogenic bacteria were analyzed. The relationship between living status and HMGB1 level was investigated. Results HMGB1 leveland APACHEⅡ scorein the survival group were significantly lower than those in the death group were[（36.26±23.57）pg/mL vs（42.67±24.86）pg/mL,16.52±6.35vs26.37±6.74,P<0.05] on the first day after admission.HMGB1 level of the survival group decreased gradually within 7 days after after admission,while HMGB1 level of the death group was unchanged. The higherHMGB1 level,the greaterinfectedrisk of Gram-negative bacteria in sepsis patients. Conclusion HMGB1 can effectively predict the progression of patients with sepsisand evaluate the survival status.

Diagnostic role of Helicobacter pylori detection in patients with peptic ulcer

HUANG Ping, WANG Jian, XU Weiqi

2021, 36(1): 84-86. DOI: 10.3969/j.issn.1673-8640.2021.01.017

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Objective To investigate the diagnostic role of Helicobacter pylori（Hp） detection in patients with peptic ulcer（PU）. Methods A total of 214 patients with PU（PU group）,146 patients with chronic superficial gastritis（CSG group）,and 118 healthy subjects（control group）were enrolled in this study. All the subjects received ¹⁴C urease breath test,and Hp antibody typing of the 3 groups were detected by immunoblotting. Results The positive rateof ¹⁴C urease breath test of PU,CSG and control groups was 85.51%（183/214）,76.03%（111/146） and 42.37%（50/118）,respectively,and there were significant differences between groups（P<0.05）. The Hp typeⅠinfectious rate of PU and CSG groups was higher than that of control group,and the Hp typeⅠinfectious rate of PU group was higher than that of CSG group（P<0.05）. The cytotoxin-associated protein（CagA）+ vacuolar protein toxin A（VacA） antibody positive rate of PU group and CSG group was higher than that of control group,and CagA+VacAantibody positive rateof PU group was higher than that of CSG group（P<0.05）. CagA+VacA antibody positive ratewas higher than single CagA or VacA antibody positive ratein both PU group and CSG group. Conclusion Hp antibody serotyping assay can provide a reliable reference for clinical diagnosis of PU and hasimportant clinical value.

Incidenceand resistance profile of carbapenem-resistant Enterobacteriaceae among inpatients in a clinical center in Shanghai from 2015 to 2019

YANG Hongmei, TAO Jianmin, WEI Tian, WANG Jiangjun, MAO Jiahong, WANG Haiying

2021, 36(1): 87-91. DOI: 10.3969/j.issn.1673-8640.2021.01.018

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Objective To analyze the incidence and resistantspectrum of carbapenem-resistant Enterobacteriaceae（CRE）in Yueyang Hospital of Integrated Traditional Chinese and Western Medicine form 2015 to 2019. Methods A total of 5 980 strains of Enterobacteriaceae were collected from various clinical specimens of inpatients from 2015 to 2019. VITEK 2 Compact 60 automatic bacterial identification system was used to identify the bacteria. Antimicrobial susceptibility testing was carried out using Kirby-Bauer method or automated system. Results A total of 923 strains（15.4%）of CRE were isolated,and the incidence of CRE showed a significant upward trend.The mean age of patients with CRE infection was（76.9±14.5）years old,and patients aged ≥60 years old accounted for 89.8%. CRE was isolated mainly from respiratory tract specimens （558 strains,60.5%）,and distributed in 30 clinical departments. In these departments, the geriatrics department had the highest detection rate （21.5%）,followed by 20.9% in the intensive care unit. Among 923 CRE isolates,carbapenem-resistant Klebsiella pneumonia was the most prevalence species（77.7%）,followed by carbapenem-resistant Escherichia coli（10.9%）. Drug sensitivity test results showed that the resistance rates of CRE to the commonly used antibiotics were above 80.0%,and the resistance rates to tigecycline,polymyxin B,amikacin,trimethoprim/sulfamethoxazole,tobramycin and gentamicin were relatively low（4.1%,5.9%,51.7%,58.4%,66.5% and 75.0%）Conclusion The CRE incidence rate of inpatients in Yueyang Hospital is on the rise from 2015 to 2019. CRE were mainlyisolatedfrom elderly patientsaged ≥ 60 years old. CRE strains involve a wide range of clinical departmentand show high resistance to most commonly used antimicrobials. Hospitals should strengthen the prevention and control of CRE infection and intelligent use of antibiotics.

Performance evaluation of acridinium ester labeled direct chemiluminescence method for the detection of AMH and establishment of adult female AMH reference interval

YUE Chaoyan, DUAN Chaohui, ZHANG Di, WEI Jie, LUO Xiaohong, ZHANG Qianlan, WU Yong, YING Chunmei

2021, 36(1): 95-100. DOI: 10.3969/j.issn.1673-8640.2021.01.020

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Objective To evaluate the performance of acridinium ester labeled direct chemiluminescence method for the detection of anti-mullerian hormone（AMH）,and to establish adult female AMH reference interval. Methods The blank limit,detection limit,quantitative limit,precision and linear range of acridinium ester labeled direct chemiluminescence method were evaluated according to the clinical and Laboratory Stabdards Institute（CLSI）documents. The consistency of electrochemiluminescence and acridinium ester labeled direct chemiluminescence method was analyzed by regression analysis and bias evaluation. The stability under different storage conditions and the difference of different specimen types were evaluated. Percentile method was used to establish a reference interval for adult female AMH. Results The blank limit was 0.001 ng/mL,the detection limit was 0.02 ng/mL,and the quantitative limit was 0.08 ng/mL. The repeatability at low concentration was 1.3%,the inter-assay precision was 2.2%,and the total imprecision was 3.2%. The repeatability at high concentration was 1.6%,the inter-assay precision was 2.1%,and the total imprecision was 3.1%. The linear interval was 0.02-27.22 ng/mL. Electrochemiluminescence and acridinium ester labeled direct chemiluminescence method had a good correlation（r=0.975 3）. There was a linear correlation between the results of serum and plasma. The serum samples were stable for 3 d at room temperature（20-25 ℃） and for 7 d at 4,-20 and -80 ℃. The reference intervals for AMH for adult females aged 20-24,25-29,30-34,35-39,40-44,45-49 and ≥50 years old were 1.20-10.21,1.14-9.17,0.55-8.18,0.25-7.02,0.07-4.59,0.01-2.11 and <0.39 ng/mL,respectively. Conclusions Acridinium ester labeled direct chemiluminescence method has good performance for detecting AMH,and the AMH reference interval of adult females can provide a reference for clinic.

Preparation of five frozen pooled human serum samples as secondary reference materials for uric acid

JIANG Yaping, XU Guobing, LIU Ou

2021, 36(1): 101-106. DOI: 10.3969/j.issn.1673-8640.2021.01.021

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Objective To prepare five frozen pooled human serum samples as secondary reference materials for uric acid（UA）,which were used to validate trueness of routine methods. Methods Five UA concentrations of fasting fresh serum samples without lipemia,hemolysisorcholoplania were collected selectively,mixed and centrifuged. The supernatants were filtrated,allotted,sealed and stored at -70 °C. The homogeneity and stability of the five frozen pooled serum samples were evaluated according to Reference Materials-General and Statistical Principles for Certification (ISO Guide 35 for short). The materials were assigned values by three candidate reference laboratories. The fixed values of the five serum samples presented as target value ± total uncertainty（u_CRM）. Commutability of the five serum samples was assessed according to the National Committee for Clinical Laboratory Standards EP14-A2. Results The five frozen pooled serum samples were homogeneous（P>0.05）,and the variation between bottles was less than 1%. The stability of UA concentration at room temperature and 4 ℃ could meet the needs of daily use. The samples were stable for at least 115 days at -20 ℃ and 15 months at -70 ℃. The fixed valuesof the five frozen pooled serum samples for UA were (70.00±3.12),(281.67±4.69),(383.00±6.46),(431.00±9.95) and (520.00±12.10) μmol/L. The five frozen pooled serum samples had good commutability among 15 routine analytical systems. Conclusion The fivefrozen pooled serum samples are homogeneous,stable,commutable and well assigned,which can be recommended as secondary reference materials for UA.

Preparation and evaluation of the quality control materials of Chlamydia trachomatis

DNA XU Mingyong, ZHU Huaqiang, WU Yonghua, ZHUANG Lidong

2021, 36(1): 107-110. DOI: 10.3969/j.issn.1673-8640.2021.01.022

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Objective To prepare quality control materials of Chlamydia trachomatis（CT）DNA,and to evaluate its stability. Methods The weak positive and positive quality control materials wereprepared with pooled secretion samples. The melting stability of the quality control materials was evaluatedby water bath at 37 ℃ and room temperature（22 ℃）. The stability of the quality control materials in freezing storage was evaluated using univariate linear regression model. Results The melting stability evaluation for the weak positive control material showed acceptable results with significant differences（P<0.05）,but positive control material showed acceptable results without significant differences（P>0.05）. The cryopreserved stability evaluation for the weak positive and positive control materials showed that no significant linear trend was found with extended test time（P>0.05）. Conclusion Positive quality control materials of CT DNA have advantages of simple preparation,good stability and low cost,and can be used in laboratory detection. The cryopreserved secretion samples in the laboratory can achieve the beststability when they are melted in 37 ℃ water bath.

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