Table of Content

30 May 2019, Volume 34 Issue 5

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Serum metabolins predicting acute myocardial infarction patients' outcome

LIU Jie, SHI Xinrong, ZHOU Shufen, XU Hongmin, ZHANG Lei, LIU Shuye

2019, 34(5):  383-389.  DOI: 10.3969/j.issn.1673-8640.2019.05.001

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Objective To find serum metabolins predicting acute myocardial infarction （AMI）patients' outcome,using the metabolomics platform of ultra-high performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS） when patients are admitted to hospital. Methods UPLC-MS/MS platform was used to analyze 48 healthy subjects（healthy control group）,48 angina patients,24 AMI patients with improvement and 24 AMI dead patients. The disease distinguishing model was established. Receiver operating characteristic（ROC） curve was used to evaluate the clinical diagnostic performance of characteristic metabolins. Results A "Normal-Angina-AMI improved-AMI dead" of the principal component analysis（PCA）（R2X=40.2%, Q2=13.9%）and orthogonal partial least-square discrimination analysis （OPLS-DA） model （R2X=79.0%, R2Y=83.2%, Q2=65.0%） were established. Through verifying,the model prediction accuracy reached 100%. Totally,10 kinds of ions that can estimate the prognosis of AMI were screened and verified. 4-Hydroxy-6-docosanone,N-acetyl-leukotriene,lysophosphatldy choline（LPC）[18： 2（9Z,12Z）],LPC（18：0）,LPC（P-16：0）,LPC[P-18：1（9Z）],LPC[20：1（11Z）],LPC[20：2（11Z,14Z）] decreased,and L-threonine and 3-methyl-2-butene-1-thiol increased. There was statistical significance between AMI patients with improvement and AMI dead patients（P<0.05）. Conclusions Specific metabolins can effectively predict the prognosis of patients with AMI. These ions may be potential prognosis biomarkers and have benefit in further clinical study.

Role of NGAL for the diagnosis of kidney injury in patients with essential hypertension

LI Bo, WANG Haiyu, LIU Yajie

2019, 34(5):  390-393.  DOI: 10.3969/j.issn.1673-8640.2019.05.002

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Objective To investigate the role of neutrophil gelatinase-associated lipocalin （NGAL） for the diagnosis of kidney injury in patients with essential hypertension. Methods A total of 154 patients with essential hypertension [stage 1 group （50 cases）,stage 2 group （47 cases） and stage 3 group （57 cases）] and 50 healthy subjects （healthy control group） were determined for the levels of serum creatinine （Cr）,cystatin C（Cys C）and NGAL. Receiver operating characteristic （ROC） curve was used to evaluate the roles of Cys C and NGAL for the diagnosis of kidney injury in patients with essential hypertension. Pearson correlation analysis was used to evaluate the correlations between variables. Results The levels of serum Cr,Cys C and NGAL in essential hypertension group were higher than those in healthy control group （P<0.05）. The estimated glomerular filtration rate（eGFR） in healthy control group was higher than those in stage 3 and 2 groups （P<0.05）. Serum NGAL,Cr and Cys C levels in stage 3 group were higher than those in stage 2 and 1 groups （P<0.05,P<0.001）. Serum NGAL level in stage 2 group was higher than that in stage 1 group （P<0.001）. The eGFR in stage 3 group was lower than those in stage 1 and 2 groups （P<0.05）. There was a strong positive correlation between serum Cr and Cys C（r=0.789,P<0.001）,but serum NGAL had a weak positive correlation with Cr（r=0.314,P<0.001）and Cys C（r=0.333,P<0.001）. The combined determination of Cys C and NGAL for the diagnosis of kidney injury in patients with essential hypertension had the area under curve（AUC） of 0.935,which was bigger than those of single determinations （that for NGAL was 0.874 and that for Cys C was 0.910）. The positive rate of combined determination （78.6%） was higher than those of Cys C and NGAL single determinations （51.2% and 69.4%,respectively,P<0.001）. Conclusions NGAL plays a role in the diagnosis of kidney injury for patients with essential hypertension.

Role of remnant lipoprotein cholesterol in the management of blood lipid

SONG Yunxiao, GE Wen, ZHANG Haichen, YUAN Wenhua, DAI Jun, YOU Wenshi

2019, 34(5):  394-400.  DOI: 10.3969/j.issn.1673-8640.2019.05.003

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Objective To investigate the role of remnant lipoprotein cholesterol （RLP-C） in the management of blood lipid and the correlation between RLP-C and cardiovascular disease（CVD） associated biomarkers. Methods Serum levels of RLP-C,glucose （Glu）,triglyceride （TG）,total cholesterol （TC）,low-density lipoprotein cholesterol （LDL-C）,high-density lipoprotein cholesterol （HDL-C）,small-density low-density lipoprotein cholesterol （sd-LDL-C）,apolipoprotein （apo） A1, apo B,lipoprotein（a） [Lp（a）],lipoprotein-associated phospholipase A₂ （Lp-PLA₂） and C-reactive protein （CRP） of 437 healthy subjects were determined. The levels of RLP-C determined directly were marked RLP-C-m, and those calculated were marked RLP-C-calc,and non-HDL-C was calculated. Correlation analysis and stratified analysis were used to investigate the correlation and discrepancy between RLP-C and other CVD associated biomarkers and the influence of methodology on the level of RLP-C acquired. Results There was statistical significance in biomarkers between females and males,except ages,Glu,LDL-C,sd-LDL-C and Lp-PLA₂ （P<0.05）. The correlations between RLP-C and ages,Glu,HDL-C,apo A1,Lp（a）,Lp-PLA₂ and CRP were relatively weak with small r （P<0.05）. RLP-C-m and RLP-C-calc were positively correlated with non-HDL-C,TC,TG,LDL-C,apo B and sd-LDL-C （P<0.001）. RLP-C-m was positively correlated with TG （r=0.899,P<0.001）. RLP-C-calc and non-HDL-C were strongly and positively correlated with TC（P<0.001）,and RLP-C-m was positively correlated with RLP-C-calc and non-HDL-C （P<0.001）. The RLP-C-m level was elevated with the increase of RLP-C-calc,but it was constantly lower than RLP-C-calc. The 437 healthy subjects were stratified according to the levels of TG （<1.00,1.00-1.99,2.00-2.99 and ≥3.00 mmol/L）. The levels of RLP-C-m,RLP-C-calc, non-HDL-C, TC,sd-LDL-C and RLP-C-m were elevated with the increase of TG. The analysis of variance（ANOVA） suggested that,among the biomarkers,RLP-C-m and RLP-C-calc showed statistical significance among all the groups （P<0.001）. Conclusions RLP-C is a constant biomarker with strong and positive correlation with TG. The levels of RLP-C determined by different methods differs from each other, but they are correlated. The determination of RLP-C might help blood lipid management and risk control in CVD patients,but further researches are required to regard it as an independent factor for predicting CVD risk.

Role of serum adiponectin determination combined with leptin in the diagnosis of lupus nephritis

XU Liming, SHEN Jun

2019, 34(5):  401-404.  DOI: 10.3969/j.issn.1673-8640.2019.05.004

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Objective To investigate the role of serum adiponectin determination combined with leptin in the diagnosis of lupus nephritis （LN）. Methods A total of 84 female patients with systemic lupus erythematosus （SLE） were enrolled and classified into LN group （38 cases） and non-LN group （46 cases） according to be with or without renal damage. Totally,45 female healthy subjects were enrolled as healthy control group. Creatinine,white blood cell count and 24 h urinary protein were determined,and serum levels of leptin and adiponectin were determined by enzyme-linked immunosorbent assay. Results Serum levels of leptin and adiponectin in LN group were （34.12±21.41） ng/mL and （21.62±9.69） μg/mL,which were higher than those in healthy control group [（17.21±16.43） ng/mL and （11.31±6.45） μg/mL] （P<0.05）. Serum level of adiponectin in LN group was higher than that in non-LN group [（15.28±5.43） μg/mL] （P<0.05）. The sensitivity and specificity of leptin determination in the diagnosis of LN were 63.16% and 54.35%,respectively. The sensitivity and specificity of adiponectin in the diagnosis of LN were 81.58% and 28.26%,respectively. The sensitivity and specificity of the combined determination of leptin and adiponectin in the diagnosis of LN were 97.37% and 19.57%,respectively. Conclusions Serum adiponectin determination combined with leptin has high sensitivity in the diagnosis of LN,and it can provide a reference for the early diagnosis of LN.

Relationship between parathyroid hormone level and renal function in patients with chronic kidney disease

XING Xiaoping, TANG Jibin, WANG Chuanfa, SUN Yun, LI Xuguang, WU Daowei, SONG Yuanming

2019, 34(5):  405-407.  DOI: 10.3969/j.issn.1673-8640.2019.05.05

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Objective To investigate the relationship between parathyroid hormone （PTH） level and renal function in patients with chronic kidney disease （CKD）. Methods A total of 149 patients with CKD were enrolled. According to the staging method of chronic renal failure in China,the CKD patients were classified into 4 groups（chronic renal insufficiency decompensated stage,chronic renal insufficiency compensated stage,chronic renal failure and uremia groups）. Totally,25 healthy subjects were enrolled as healthy control group. PTH levels and biochemical indexes were compared and analyzed,and correlation analysis was conducted. Results Serum PTH,blood urea nitrogen （BUN）,creatinine（Cr）,cystatin C（Cys C） and phosphorus（P） levels in CKD group were higher than those in healthy control group （P<0.05）,and they were gradually increased with CKD stages （P<0.05）. There was no statistical significance for serum calcium（Ca）（P>0.05）. Pearson correlation analysis showed that PTH was positively correlated with BUN,Cr,Cys C and P（r=0.422,0.616,0563 and 0.376,P<0.01）,and it was negatively correlated with Ca（r=-0.332,P<0.01）. Conclusions PTH could reflect the progress of CKD to some extent. Monitoring serum PTH level of patients with chronic renal failure may be helpful to understand the disease progress and evaluate the disease severity.

Role of the avidity of Epstein-Barr virus capsid antigen-specific IgG antibodies and immunological changes among children with infectious mononucleosis

ZHENG Lan, CHENG Juan, PAN Qiuhui, YANG Lin

2019, 34(5):  408-414.  DOI: 10.3969/j.issn.1673-8640.2019.05.006

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Objective To evaluate the role of the avidity of Epstein-Barr virus （EBV）capsid antigen（EBV-CA） -specific IgG antibodies and immunological changes among children with infectious mononucleosis （IM）. Methods A total of 5 882 children （500 cases of IM and 5 382 cases of non-IM） were enrolled for determining the avidities of EBV-CA IgG antibodies and EBV-CA IgM antibodies. The avidities of EBV-CA IgG antibodies and EBV-CA IgM antibodies,EBV DNA load,heterotypic lymphocytes and lymphocyte subsets in peripheral blood were determined. Receiver operating characteristic （ROC） curves were used to evaluate the efficiency of the avidities of EBV-CA IgG antibodies and EBV-CA IgM antibodies,heterotypic lymphocyte percentage and EBV DNA individually and combinedly in the diagnosis of IM. The immunological changes among children with IM were observed. Totally,502 clinically assigned IM child samples conducted EBV DNA and lymphocyte subset determinations were reviewed as well,and the immunological changes were compared between EBV DNA positive and negative samples. Results A total of 1 031 of 5 882 （17.53%） children got any one of the 4 indicators with the avidities of EBV-CA IgG antibodies and EBV-CA IgM antibodies,heterotypic lymphocyte percentage and EBV DNA as a positive control. IM ranked top （38.1%） in the disease panel. Receiver operating characteristic （ROC） curve analysis revealed that the areas under curve （AUC） for the avidities of EBV-CA IgG antibodies and EBV-CA IgM antibodies,heterotypic lymphocyte percentage and EBV DNA reached to 0.883,0.729,0.788 and 0.664,respectively,while the 4 indicators' combined analysis got the AUC of 0.847. The 1 031 children were classified into 4 groups as infant （<1-year-old）,baby （1-3-year-old）,preschool age （4-6-year-old） and school age （7-17-year-old） groups. Except infant group,in the other 3 groups,IM patients had lower proportions of CD3^-CD16⁺CD56⁺ natural killer（NK） cells,CD3^-CD19⁺B cells,CD3⁺CD4⁺ T cells and CD4⁺/CD8⁺ ratio than those of non-IM patients （P<0.01）,and they had higher proportions of CD3⁺ T cells and CD3⁺CD8⁺ T cells than those of non-IM patients （P<0.01）. In infant group,IM patients had lower CD3^-CD19⁺ B cell proportion （P<0.01） and higher CD3⁺ T cell,CD3⁺CD8⁺T cell proportions than non-IM patients （P<0.01）. There was no statistical significance of CD3^-CD16⁺CD56⁺ NK cell proportion,CD3⁺CD4⁺ T cell proportion and CD4⁺/CD8⁺ ratio between IM and non-IM patients （P>0.05）. In IM patients,the alteration of lymphocyte subsets got more significance in EBV DNA positive patients than negative patients. Conclusions For the diagnosis of IM,the avidity of EBV-CA IgG antibody is more efficient than heterotypic lymphocyte percentage,EBV-CA IgM antibody and EBV DNA. Cell immune status is disturbed in IM children,and is even worse for those with positive EBV DNA.

Difference of the expressions of ER,VEGF and TGF-β1 between multiple EP and single EP patients and their prognosis analysis

DONG Huan, LIU Xiaoyi, CAI Yanqing, CHEN Xinliang, WANG Xianjing

2019, 34(5):  415-420.  DOI: 10.3969/j.issn.1673-8640.2019.05.007

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Objective To investigate the expression levels of estrogen receptor （ER）,vascular endothelial growth factor （VEGF）and transforming growth factor-beta 1（TGF-β1）in multiple endometrial polyp（EP）and single EP patients,and to analyze the relationship with prognosis. Methods A total of 100 premenopausal women undergoing hysteroscopic endometrial polypectomy were recruited. Among them,there were 50 multiple EP （>5） patients and 50 single EP patients. Totally,35 patients undergoing hysteroscopy due to oviductal problem were enrolled as normal endometrium group. Immunohistochemistry and reverse transcription polymerase chain reaction （RT-PCR） were used to determine the expression levels of ER,VEGF and TGF-β1 in normal endometrium,single EP and multiple EP groups. Kaplan-Meier curve was used to analyze the recurrence rate of EP patients. Cox regression analysis was used to assess the risk of EP recurrence. Results The mRNA and protein expression levels of ER,VEGF and TGF-β1 in the polyp tissues of multiple EP group and single EP group were higher than those in normal endometrium group （P<0.05）,and they were higher in multiple EP group than those in single EP group （P<0.05）. The recurrence rate in multiple EP group （42.0%） was higher than that in single EP group （12.0%） （P=0.001）. Cox regression analysis showed that endometriosis [hazard ratio（HR）=2.718,95% confidence interval（CI） 1.179-6.262] and multiple EP （HR=4.134,95%CI 1.663-10.273） were independent risk factors for EP recurrence,and the high expressions of VEGF （HR=1.073, 95%CI 1.014-1.136） and TGF-β1 （HR=1.056,95%CI 1.005-1.109）were independent risk factors for recurrence in multiple EP patients. Conclusions The high expressions of ER,VEGF and TGF-β1 in multiple EP group are closely related to the high recurrence rate after operation.

Drug resistance of 218 isolates of Mycobacterium tuberculosis in

Xi'an YANG Jian, ZHANG Tianhua, XIAN Xiaoping, LI Yan, CHEN Meiling, WANG Rui

2019, 34(5):  421-427.  DOI: 10.3969/j.issn.1673-8640.2019.05.008

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Objective To analyze the drug resistance and risk factors of 218 isolates of Mycobacterium tuberculosis （MTB） in Xi'an. Methods A total of 218 isolates of MTB from 227 smear positive tuberculosis patients' sputum samples were collected in a surveillance point of Xi'an. The proportional method was used for the drug susceptibility test of 9 anti-tuberculosis drugs. Multivariate Logistic regression analysis was used to analyze the risk factors. Results The overall drug resistance rate was 32.57% （29.03% for the initial treatment cases and 53.13% for the retreatment cases）. The multi-drug resistance rate was 6.88%. The drug resistance rates to first-line and second-line anti-tuberculosis drugs were 25.69% and 15.14%,respectively. It showed the highest resistance to streptomycin （SM）,followed by isoniazid （INH）,rifampicin （RFP）,ethambutol （EMB）,ofloxacin （OFX）,capreomycin （CM）,p-aminosalicylic acid（PAS）,kanamycin （KM） and protionamide （PTO）. There were 29 types of drug resistance,including 6 cases of mono-drug resistance,3 cases of poly-drug resistance,7 cases of multi-drug resistance,11 cases of second-line drug resistance and 2 cases of other drug resistance. The drug resistance rate and multi-drug resistance rate in retreatment group were higher than those in initial treatment group [odds ratio （OR） =3.39,95% confidence interval（CI）1.49-7.70,χ²=8.52,P=0.004;OR=4.83,95%CI 1.53-15.25,χ²=7.21,P=0.01]. The multi-drug resistance rate in the group of annual family income less than 40 thousand was higher than that of corresponding control group （OR=3.96,95%CI 1.31-11.97,χ²=5.94,P=0.02）. Conclusions The overall drug resistance rate and multi-drug resistance rate of pulmonary tuberculosis patients in Xi'an are lower than those of the whole nation. The drug resistant spectrum has polymorphism and complexity. The drug retreatment patients are more likely to have drug resistance and multi-drug resistance,and low family income patients are more likely to have multi-drug resistance.

Activities of tedizolid in vitro against clinical Gram-positive cocci

LIN Wen, HUANG Kaifeng, YANG Han, HE Chunyan, CHEN Wenjing, SHU Wen, TANG Rong, YI Junwen, LIU Qingzhong

2019, 34(5):  428-432.  DOI: 10.3969/j.issn.1673-8640.2019.05.009

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Objective To investigate the drug susceptibilities of a novel oxazolidinone antibiotic,tedizolid,against clinical Gram-positive cocci,and to investigate the drug resistance mechanism in non-tedizolid susceptible isolates. Methods A total of 1 069 clinical Gram-positive cocci were collected. The isolates included 202 isolates of methicillin-resistant Staphylococcus aureus （MRSA）,294 isolates of methicillin-susceptible Staphylococcus aureus （MSSA）,115 isolates of coagulase-negative Staphylococcus,206 isolates of Enterococcus faecalis,55 isolates of Enterococcus faecium,159 isolates of Streptococcus agalactiae and 38 isolates of Streptococcus anginosus. The minimum inhibitory concentrations （MIC） of tedizolid and linezolid were determined by broth microdilution method and analyzed to clarify the difference of in vitro activities of them. Polymerase chain reaction （PCR） was performed to determine the drug resistance genes in tedizolid/linezolid non-susceptible isolates. Results All isolates of Staphylococcus （MIC≤0.5 μg/mL）,Enterococcus faecium （MIC≤0.5 μg/mL） and Streptococcus （MIC≤0.25 μg/mL） were susceptible to tedizolid. The susceptibility rate of Enterococcus faecalis to tedizolid was 94.7%. Totally,11 tedizolid and linezolid non-susceptible isolates （tedizolid MIC=1 μg/mL,linezolid MIC=8 μg/mL） were found,which were positive for the novel oxazolidinone resistance gene, optrA. Tedizolid revealed 4-8-fold more effective than linezolid. Conclusions Tedizolid is a novel antibiotic for treating Gram-positive cocci infection,however,its drug resistance mediated by optrA in Enterococcus should be concerned.

Roles of PCT,HNL and CD64 in the diagnosis of bacterial infection

MA Caiyan, LAI Wenqiang, CHEN Xiaoping

2019, 34(5):  433-436.  DOI: 10.3969/j.issn.1673-8640.2019.05.010

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Objective To investigate the roles of procalcitonin （PCT）,human neutrophil lipocalin （HNL）and neutrophil CD64 in the diagnosis of bacterial infection. Methods A total of 86 patients with suspected bacterial infection were enrolled. There were 44 cases of bacterial infection group,42 cases of viral infection group and 40 cases of healthy subjects（healthy control group）. The levels of PCT,HNL,CD64 and white blood cell （WBC） count of bacterial infection,viral infection and healthy control groups were determined,and the diagnostic roles of PCT,HNL and CD64 were evaluated by receiver operating characteristic（ROC）curve. Results Compared with viral infection and healthy control groups,PCT,HNL and CD64 levels and WBC counts in bacterial infection group were increased（P<0.05）. CD64 level and WBC count were increased in viral infection group compared with healthy control group（P<0.05）. The sensitivities of PCT,HNL,CD64 and WBC count for the diagnosis of bacterial infection were 79.55%,84.09%,81.82% and 77.27%,respectively. The specificities were 83.33%,88.09%,85.71% and 85.71%,respectively. The areas under ROC curves （AUC） of HNL,CD64,PCT and WBC count were 0.914,0.900,0.837 and 0.748,respectively. Conclusions HNL,CD64,PCT and WBC count can be used in the diagnosis of bacterial infection. HNL plays a role for the diagnosis of bacterial infection.

Reference intervals of venous blood cell parameters of healthy Tibetan adults in Hongyuan county

LIU Guoxiong, LIU Yong, LIU Yunli, ZHANG Liwen, XIE Xinyue

2019, 34(5):  437-440.  DOI: 10.3969/j.issn.1673-8640.2019.05.011

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Objective To establish the reference intervals of venous blood cell parameters of healthy Tibetan adults in Hongyuan county,Aba prefecture,Sichuan province. Methods A total of 1 150 healthy Tibetan adults of 16-60 years old were enrolled from 11 towns and villages of Hongyuan county for generations. There were 648 males and 502 females. Venous blood cell parameters [red blood cell （RBC）count,hemoglobin （Hb）,white blood cell （WBC） count and platelet （PLT） count] were determined by XS-800i automatic blood cell analyzer. The results were compared. The reference intervals were eatablished as x±1.96s. Results WBC counts were （3.47-11.89）×10⁹/ L for males and （3.68-11.02）×10⁹/L for females. RBC counts were （4.81-6.57）×10¹²/L for males and （4.17-5.89）×10¹²/ L for females. Hb levels were 158-209 g/L for males and 129-186 g/L for females. PLT counts were （89-274）×10⁹/L for males and （98-312）×10⁹/L for females. The reference intervals had been established and were compared with the reference intervals of the 4th edition of the current national practice for clinical examination the National Guide to Clinical Laboratory Procedures,showing statistical significance （P<0.05）. Conclusions The regional reference intervals of venous blood cell parameters of healthy Tibetan adults in Hongyuan county have been established.

IFCC Secondary Reference Measurement Procedure for 4 HbA_1c determination systems

LIU Wenbin, JU Yi, TANG Liping, WANG Meijuan, OU Yuanzhu, YU Xiaoxuan, LI Qing

2019, 34(5):  443-446.  DOI: 10.3969/j.issn.1673-8640.2019.05.013

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Objective To improve the traceability of glycated hemoglobin A_1c（HbA_1c） in Shanghai Center for Clinical Laboratory with establishing the International Federation of Clinical Chemistry and Laboratory Medicine （IFCC） Secondary Reference Measurement Procedure,through tracing from the IFCC Primary Reference System. Methods Using 4 HbA_1c determination systems,including HA-8180,Hb-9210,Capillarys 2 Flex Piercing （Cap2） and MQ2000-PT with their original reagents and calibrators,the results of 24 samples sent by IFCC were compared with the target values assigned by the IFCC Primary Reference System. The performance of the 4 determination systems were evaluated by the IFCC rules. Results The coefficients of variation （CV） of the 4 determination systems were 1.1% for HA-8180,2.0% for Hb-9210,2.2% for Cap2 and 2.0% for MQ2000-PT. The imprecisions were 0.6,1.0,1.1 and 1.0 mmol/mol,respectively. The |biases| at the level of 50 mmol/mol were 1.0,1.1,0.4 and 1.0 mmol/mol,respectively. The total errors （TE） of whole year were 2.2,3.1,2.6 and 3.2 mmol/mol,respectively. According to the rules,HA-8180 got a "Sliver" score,and Hb-9210,Cap2 and MQ2000-PT got "Bronze" score. Conclusions The 4 determination systems demonstrate traceability to the IFCC Secondary Reference Measurement Procedure.

Performance evaluation for the direct detection of pathogenic bacteria causing urinary tract infection by

MALDI-TOF MS SHEN Zhenhua, CHEN Rong

2019, 34(5):  447-450.  DOI: 10.3969/j.issn.1673-8640.2019.05.014

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Objective To investigate the role of matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF MS） in the detection of pathogenic bacteria causing urinary tract infection. Methods A total of 1 000 midstream urine samples were collected and inoculated on solid media. Single colonies were identified by MALDI-TOF MS and Vitek 2 Compact. Midstream urine samples were directly detected by MALDI-TOF MS after pretreatment. Results Among the 1 000 midstream urine samples,215 cases with colony count ≥10⁵ CFU/mL were detected by culture method,and the positive detection rate was 21.5%. Among them,there were 162 cases isolated with single bacterium and 53 cases with mixed bacteria. The consistency rate for the identification of 215 cases by MALDI-TOF MS and Vitek 2 Compact was 100 %. Gram-negative bacteria accounted for 76.3%. Gram-positive bacteria accounted for 23.7%. Totally,185 cases with colony count ≥10⁵ CFU/mL were detected by MALDI-TOF MS directly,and the positive detection rate was 18.5%. Among them,145 cases of single bacterium were isolated,and 40 cases of mixed bacteria were found. The direct detection method had no false positive case. The positive detection rate was lower than that of culture method,and the positive leakage rate reached 14.0 %（30/215）. Conclusions MALDI-TOF MS for the direct detection of pathogenic bacteria from infectious midstream urine samples has the advantages of short identification and diagnosis time, low cost and good applicability.

ATRA for promoting brown adipogenesis in C3H10T1/2 mesenchymal stem cells

LIANG Qiuli, CHENG Jinke, ZUO Yong

2019, 34(5):  451-456.  DOI: 10.3969/j.issn.1673-8640.2019.05.015

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Objective To investigate the effect and mechanism of all-trans retinoic acid （ATRA） or bone morphogenetic protein 7 （BMP7） on the development of brown adipocytes in C3H10T1/2 mesenchymal stem cells. Methods C3H10T1/2 mesenchymal stem cells were pretreated with ATRA or BMP7,followed by an adipogenic cocktail. To evaluate the adipogenesis,oil red O staining was used to identify brown adipogenic capacity. Mitochondrial track staining was used to analyze mitochondrial biogenesis in mature brown adipocytes. The expressions of BMP7,brown adipogenic inhibitors （Necdin,Wnt 10a and Pref 1） and brown adipocyte-related gene were determined by real-time fluorescence quantitation polymerase chain reaction （PCR）and western blotting. Results The optimal concentration of ATRA in promoting C3H10T1/2 brown adipogenesis was 1-2 μmol/ L. After pretreatment for 3 d with 1 μmol/L ATRA （ATRA group）,the mitochondrial biogenesis gene mRNA expressions of CyctoC,Tfam and Nrf1 were increased （P<0.05）,the mRNA expressions of brown adipogenic inhibitor Necdin and its downstream target genes （Pref1 and Wnt10a） were decreased （P<0.05）,and the mRNA expression of BMP7 gene was increased （P<0.05）,compared with control group treated by dimethyl sulfoxide （DMSO）. Different concentrations of ATRA can induce BMP7 protein expression in a dose-response manner. After the pretreatment with 1 μmol/L ATRA for 0,24 and 48 h,BMP7 protein expression was increased,and Necdin protein expression was decreased. The differentiation effect of ATRA and BMP7 was comparable. Conclusions ATRA promotes brown adipogenesis of C3H10T1/2 mesenchymal stem cells via the activation of BMP7 and reduces negative modulators of brown preadipocyte differentiation.

Development and performance evaluation of the national standard materials of lithium,sodium,potassium,magnesium,calcium and chlorine in compound frozen human serum

YU Ting, SHEN Min, QU Shoufang, HUANG Jie

2019, 34(5):  457-462.  DOI: 10.3969/j.issn.1673-8640.2019.05.016

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Objective To develop the national standard materials of lithium,sodium,potassium,magnesium,calcium and chlorine in compound frozen human serum and evaluate their performance,and to promote the standardization of lithium,sodium,potassium,magnesium,calcium and chlorine assays. Methods Serum samples without hemolysis,lipemia and choleplania were collected. After multiple filtration sterilization,serum pools were packed in ampoules in 3 levels' candidates and stored in -70 ℃. Potassium ion is used as a characteristic parameter for homogeneity evaluation by single factor analysis of variance. Potassium,sodium and chlorine ions were used as the characteristic parameters for stability evaluation by linear regression analysis. The value was assigned by reference methods or validated methods,and the uncertainty was calculated. The commutability of fresh serum samples between reference methods/validated methods and 4 conventional analysis systems was evaluated. Results The F values of homogeneity for potassium ion in 3 levels' candidates were 1.630 1,1.498 6 and 1.020 8,respectively. All of them were <F_0.05. The stabilities of potassium, sodium and chlorine ions can be last at least 30 d at -20 ℃,2-8 ℃ and 20-25 ℃. The assigned values for lithium,sodium,potassium,magnesium,calcium and chlorine ions were：lithium 0.649±0.022,1.195±0.032 and （1.334±0.008） mmol/L;sodium 120.057±2.206,142.030±1.521 and （158.228±1.293） mmol/L;potassium 5.042±0.143,5.739±0.106 and （6.158±0.094） mmol/L;magnesium 0.732±0.021,0.857±0.026 and （1.144±0.026） mmol/L;calcium 2.014±0.078,2.348±0.056 and （3.070±0.024） mmol/L;chlorine 90.814±2.005,108.875±0.908 and （128.156±1.453） mmol/L（k=2）. The linearities for sodium,potassium,magnesium,calcium and chlorine ions were all within 95% confidence intervals,and the commutability was good. Conclusions The homogeneity and stability of candidates all meet the requirement,and their values are assigned accurately and reliably. The commutabilities of sodium,potassium,magnesium,calcium and chlorine ions in candidates are good. They can be used as national standard materials.

Progress in the clinical application of monoclonal antibody drugs

XIAO Xiao, FENG Huijuan, GAO Chunfang

2019, 34(5):  466-471.  DOI: 10.3969/j.issn.1673-8640.2019.05.018

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Antibody is a large and Y-shaped protein that is secreted by effect B cells and binding to antigen specificity. Monoclonal antibody is a highly homogeneous antibody against a specific antigen epitope produced by single B cell clone. Because of its high specificity,monoclonal antibody is widely used in the diagnosis and treatment of diseases. In 1990s,polymerase chain reaction technology,antibody library technology and transgenic technology promoted the development of monoclonal antibody preparation technology to all humans. Monoclonal antibody has a broad application prospects in disease diagnosis and treatment of tumors and autoimmune infectious diseases. This review focuses on the application of monoclonal antibody drugs in clinical medicine.