Table of Content

28 February 2019, Volume 34 Issue 2

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Research improvement of the basis and transformation of tumor markers in clinical laboratories

GAO Feng

2019, 34(2):  95-97.  DOI: 10.3969/j.issn.1673-8640.2019.02.001

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Due to the lack of specific and sensitive tumor markers,especially early tumor markers,the overall 5-year survival rate of tumor patients is low. However,the research progress of tumor markers is slow,and the clinical application is limited. It is urgent for clinical laboratories to strengthen the research on the basis and transformation of tumor markers for developing specific and sensitive tumor markers. This paper summarizes 5 papers published in the issue,hoping that the development and application of tumor markers could be promoted.

Research progress of COPS5 in tumors

CHEN Miaomiao, GUO Lin, LU Renquan

2019, 34(2):  98-102.  DOI: 10.3969/j.issn.1673-8640.2019.02.002

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Constitutively photomorphogenic 9 signalosome subunit 5（COPS5） is an evolutionarily conserved and multifunctional protein that plays a role in signal transduction,cell proliferation,cell apoptosis,cell cycle and the regulation of genomic instability and DNA damage response. COPS5 contributes to oncogenesis by functionally inactivating key negative regulatory proteins and tumor suppressors（P53,P27 and so on）,and it is associated with poor prognosis and chemo-radiotherapy tolerance. The small molecule inhibitors for COPS5 have been developed in the preliminary clinical application. Furthermore,COPS5 may serve as a potential therapeutic target and plays a role in tumor treatment. This review focuses on the function of COPS5 in oncogenesis and the research progress of COPS5 signaling pathways.

Role of TFCP2/YAP transcriptional complex co-regulated protein CPE for the diagnosis of hepatocellular carcinoma

FANG Zixiang, ZHANG Xiao, CHEN Yan, WU Qi, ZHU Guoqing, LIU Ya, WANG Jiayi, SUN Fenyong, YU Lei

2019, 34(2):  103-109.  DOI: 10.3969/j.issn.1673-8640.2019.02.003

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Objective To investigate the role of transcription factor CP2（TFCP2）/Yes-associated protein（YAP） transcriptional complex co-regulated protein carboxypeptidase E（CPE） for the diagnosis of hepatocellular carcinoma（HCC）. Methods The levels of serum CPE in 147 HCC patients,42 hepatitis B patients,29 gastric cancer patients and 100 healthy subjects（healthy control group） were determined by enzyme-linked immunosorbent assay. Real-time fluorescence quantitation polymerase chain reaction（PCR） was used to determine the expression of CPE mRNA. Western blotting was used to determine the expression of CPE,TFCP2 and YAP. The dual-luciferase reporter test was used to determine the activity of CPE promoter. The correlations between CPE and other indicators such as alpha-fetoprotein（AFP）,alanine aminotransferase（ALT） and aspartate aminotransferase（AST） were analyzed using Spearman correlation analysis. Receiver operating characteristic（ROC） curve was used to compare the efficiency of CPE and AFP for the diagnosis of HCC. Results The overexpression of YAP or TFCP2 stimulated the expression of CPE mRNA and protein,and the overexpression of YAP and TFCP2 stimulated the expression ofCPE mRNA and protein. The knockdown of YAP or TFCP2 inhibited the expression of CPE mRNA and protein,and the knockdown of YAP and TFCP2 inhibited the expression of CPE mRNA and protein. A TFCP2-binding-motif was existed in CPE promoter region. After mutation,TFCP2 and YAP lost the regulation function on CPE promoter. The level of serum CPE was higher in HCC group than those in healthy control,hepatitis B and gastric cancer groups（P<0.01）,and there was no statistical significance among healthy control,hepatitis B and gastric cancer groups（P>0.05）. The level of serum CPE was positively correlated with the levels of AFP,ALT and AST（r=0.644,0.454 and 0.351,P=0.000,0.000 and 0.001,respectively）. The area under ROC curve（AUC） of CPE for the diagnosis of HCC was 0.937,which is better than that of AFP（0.679）. Conclusions CPE plays a role in the diagnosis of HCC,and CPE may be a diagnostic biomarker for HCC.

Preliminary study on the mechanism of CD4⁺CD25^highCD127^low regulatory T cells promoting the metastasis of epithelial ovarian cancer

KE Xing, ZHANG Liang, SHEN Lisong

2019, 34(2):  110-115.  DOI: 10.3969/j.issn.1673-8640.2019.02.004

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Objective To investigate the role of CD4⁺CD25^highCD127^low regulatory T cells（Treg） in the metastasis of epithelial ovarian cancer（EOC）,and to analyze the signaling mechanism of the related cytokines. Methods Flow cytometry was used to determine the percentages of CD4⁺CD25^highCD127^low Treg in peripheral blood of 34 patients with EOC,26 patients with benign ovarian tumors and 34 healthy subjects（healthy control group）. The clinical data of all patients,including age,pathological type,the International Federation of Gynecology and Obstetrics（FIGO） stage,histological differentiation and lymph node metastasis,were collected. A coculture system between EOC cell line OVCAR3 and healthy subjects' CD4⁺ T cells was established. The levels of interleukin 10（IL-10） and transforming growth factor-beta 1（TGF-β1） were determined by enzyme-linked immunosorbent assay（ELISA）. The expression levels of matrix metalloproteinase（MMP）-2 mRNA and tissue inhibitor of metalloproteinase（TIMP）-2 mRNA in OVCAR3 were determined by real-time fluorescence quantitation polymerase chain reaction（PCR）. The levels of MMP-2 mRNA and TIMP-2 mRNA were also analyzed after blocking IL-10 receptor and TGF-β1 receptor in OVCAR3 after the coculture experiment. Results The level of CD4⁺CD25^highCD127^low Treg in EOC group was higher than those in benign ovarian tumor and healthy control groups（P<0.05）,and there was no statistical significance between benign ovarian tumor and healthy control groups（P>0.05）. The percentage of CD4⁺CD25^highCD127^low Treg was higher in lymphatic invasion group than that in non-lymphatic invasion group（P<0.05）. There was no statistical significance for CD4⁺CD25^highCD127^low Treg percentage in EOC group with different ages,pathological types,FIGO stages and histological differentiation（P>0.05）. The levels of IL-10 and TGF-β1 from CD4⁺ T cells were up-regulated after coculturing with CD4⁺ T cells（P<0.05）. The level of MMP-2 mRNA was up-regulated（P<0.05）,and the level of TIMP-2 mRNA in OVCAR3 was down-regulated in the coculture experiment with CD4⁺ T cells（P<0.05）. The level of MMP-2 mRNA was down-regulated,and the level of TIMP-2 mRNA in OVCAR3 was up-regulated after blocking（P<0.05）. Conclusions CD4⁺CD25^highCD127^low Treg can enhance the expression of IL-10 and TGF-β1 and stimulate the related receptors on EOC cells. It plays a role in the invasion and metastasis of EOC.

Combined determination of plasma hyaluronan and the ratio of M2 monocyte/M1 monocyte ratio in breast cancer

ZHANG Boke, HUANG Yi, LIU Yiwen, HE Yiqing, YANG Cuixia, DU Yan, ZHANG Guoliang, GAO Feng

2019, 34(2):  116-121.  DOI: 10.3969/j.issn.1673-8640.2019.02.005

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Objective To investigate the role of the combined determination of plasma hyaluronan（HA） and the ratio of M2 monocyte/M1 monocyte（M2/M1 ratio） in breast cancer. Methods A total of 98 patients with breast cancer（breast cancer group）,37 patients with breast benign diseases（breast benign disease group） and 41 healthy subjects（healthy control group） were enrolled. Plasma HA and carbohydrate antigen（CA）15-3 levels were determined by chemiluminescence. Flow cytometry was used to assess the expression of M1（CD14⁺CD163^-CD86⁺） and M2（CD14⁺CD163⁺CD204⁺） monocytes,and M2/M1 ratio was calculated. Correlation analysis was performed using Spearman correlation analysis. Receiver operating characteristic（ROC） curve was used to analyze the diagnostic efficiency of each marker. Results Plasma HA and CA15-3 levels,the proportion of M2 monocyte and M2/M1 ratio were higher in breast cancer group than those in breast benign disease and healthy control groups（P<0.05）,and there was no statistical significance between breast benign disease and healthy control groups（P>0.05）. There were correlations of HA with M2/M1 ratio and the proportions of M2 monocyte and M1 monocyte（r_s=0.401,0.393 and-0.189,P<0.05）. The area under ROC curve（AUC）,sensitivity and specificity of M2/M1 ratio for the diagnosis of breast cancer were higher than those of the other markers,and the combined determination of HA and M2/M1 ratio could improve the diagnostic efficiency of breast cancer. Plasma HA was correlated with the histological differentiation of breast cancer（P<0.05）,and M2/M1 ratio was correlated with tumor-node-metastasis（TNM） stage,histological differentiation,lymph node metastasis and estrogen receptor（ER）（P<0.05）. However,there was no relationship between CA15-3 and clinicopathological characteristics（P>0.05）. Conclusions The combined determination of HA and M2/M1 ratio plays a role in the auxiliary diagnosis of breast cancer.

Role of serum CgA determination for neuroendocrine neoplasma

WU Jun, LU Renquan, GUO Lin

2019, 34(2):  122-125.  DOI: 10.3969/j.issn.1673-8640.2019.02.006

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Objective To investigate the role of serum chromogranin A（CgA） determination for neuroendocrine neoplasma（NEN）. Methods In 63 patients with NEN（NEN group）,16 patients with gastrointestinal and pancreatic tumors（digestive system tumor group） and 30 healthy subjects（healthy control group） Serum CgA levels and carbohydrate antigen（CA）19-9 levels were determined. The clinicopathological characteristics（sex,age,the primary site of tumors,the grade of tumors and the presence or absence of distant metastasis） of NEN patients were recorded. The efficiency of serum CgA for the diagnosis of NEN was evaluated by receiver operating characteristic（ROC）curve. Results Serum CgA level in NEN group was higher than those in digestive system tumor group and healthy control group（P<0.05）,and there was no statistical significance between digestive system tumor group and healthy control group（P>0.05）. Serum CgA level in NEN patients with distant metastasis was higher than that without distant metastasis（P<0.05）. There was no statistical significance for serum CgA levels in patients with different sex,ages,the primary sites of tumors and the grades of tumors（P>0.05）. ROC curve analysis showed that the area under curve（AUC）of serum CgA in the diagnosis of NEN was 0.989,the optimal cut-off value was 47.31 ng/mL,the sensitivity was 95.5%,and the specificity was 92.3%. Using digestive system tumor group as control,the AUC of serum CgA for the differential diagnosis of NEN was 0.948,the optimal cut-off value was 50.97 ng/mL,the sensitivity was 84.8%,and the specificity was 90.9%. The AUC of serum CgA combined with CA19-9 was 0.956,the sensitivity was 90.5%,and the specificity was 81.8%. Compared with the results of preoperation,serum CgA level decreased in 25 of 32 NEN patients after operation（P<0.05）. There was no statistical significance for serum CgA level before and after operation in the 7 patients（P<0.05）.Conclusions CgA can be used for the assistant diagnosis and therapeutic monitoring of NEN,and it plays a role in predicting tumor metastasis.

Influence of CYP2C9 and VKORC1 gene polymorphisms on warfarin anticoagulation strategy after cardiac valve replacement

HAO Yexia, ZHENG Xuan, HU Yuanping, ZHANG Litao, YAN Xinsheng, CAO Shuzheng, WANG Zhang, ZHANG Zhenlu

2019, 34(2):  126-129.  DOI: 10.3969/j.issn.1673-8640.2019.02.007

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Objective To evaluate the influence of CYP2C9 and VKORC1 gene polymorphisms on warfarin anticoagulation strategy after cardiac valve replacement. Methods A total of 380 patients undergoing cardiac valve replacement were enrolled randomly,CYP2C9 and VKORC1 gene polymorphisms in 190 cases were detected（gene polymorphism detection group）,and the other 190 cases were used as control group. The warfarin stable dose,the time to reach therapeutic international normalized ratio（INR） and hospitalization time were recorded. Results There were 170 cases of *1*1（89.5%）,19 cases of *1*3（10.0%） and 1 case of *3*3（0.5%） in CYP2C9,and 155 cases（81.6%） had AA,and 35 cases（18.4%） had AG in VKORC1. There was no case with GG. The time to reach therapeutic INR of gene polymorphism detection group [（4.0±1.7）d] was shorter than that of control group [（4.6±2.1）d]（P<0.01）. There was no statistical significance for hospitalization time between the 2 groups（P>0.05）. Conclusions CYP2C9 and VKORC1 gene polymorphism detection plays a role for early clinical anticoagulation strategy,which can shorten the time to reach therapeutic INR and reduce hemorrhage and embolism risk.

Role of plasma aldosterone to direct renin ratio in primary aldosteronism

WU Bing, YU Fang, ZHANG Yong, ZHOU Yuntao, SHI Lu, TIAN Wei

2019, 34(2):  130-135.  DOI: 10.3969/j.issn.1673-8640.2019.02.008

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Objective To investigate the role of plasma aldosterone to direct renin ratio（ADRR） in the diagnosis of primary aldosteronism（PA） and its optimal cut-off value. Methods A total of 222 hypertension patients were enrolled,and there were 33 cases of PA. Plasma aldosterone and renin were determined by chemiluminescence immunoassay. ADRR was calculated. Using receiver operating characteristic（ROC） curve,the role and cut-off value of ADRR for the diagnosis of PA were evaluated. Results The proportion of PA in hypertension patients was 14.86%（33/222）. Plasma renin levels at both supine and upright positions in PA group were lower than those in hypertension group（P<0.05）. ADRR in PA group was higher than that in hypertension group（P<0.05）. ROC curve analysis showed that the areas under curves（AUC） of ADRR were 0.906（supine position）and 0.908（upright position）. The optimal cut-off value of ADRR at supine position was 31.32. The sensitivity was 96.97%,and the specificity was 73.54%. However,the optimal cut-off value of ADRR at upright position was 27.57,the sensitivity was 81.82%,and the specificity was 86.77%. Conclusions Plasma ADRR is an effective index for PA screening,and the cut-off value 25 is optimal for screening ADRR at upright position.

Virulence genes and drug resistance of Vibrio parahaemolyticus isolated from patients with diarrhea in Ningbo

XIE Hongyi, YE Hongyan, ZHOU Fangman, SHI Lumei, GE Tingting, LE Xubo

2019, 34(2):  136-139.  DOI: 10.3969/j.issn.1673-8640.2019.02.009

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Objective To investigate the virulence genes,type Ⅲ secretion system（T3SS） and drug resistance of Vibrio parahaemolyticus isolated from patients with diarrhea in Ningbo. Methods A total of 49 isolates of Vibrio parahaemolyticus were isolated from feces specimens of patients with diarrhea. The virulence genes,tdh,trh and T3SS1,T3SS2,were determined by polymerase chain reaction（PCR）. The drug susceptibility of 17 kinds of antibiotics were determined by disk diffusion assay. Results Among the 49 isolates of Vibrio parahaemolyticus,48 isolates carried tdh. There was 1 isolate carrying trh,47 isolates showed tdh（+）trh（-）,1 isolate showed tdh（+）trh（+）,and 1 isolate showed tdh（-）trh（-）. The 49 isolates carried T3SS1 gene,47 isolates carried T3SS2α gene,and 1 isolate carried T3SS2β gene. The drug resistance rate to ampicillin was 75.5%. The drug susceptibility rates were high to the 3rd generation cephalosporins and aminoglycoside tetracyclines. All the isolates were sensitive to cefepimes,levofloxacins and carbapenems（imipenem and meropenem）,and the drug susceptibility rates were 100%. Conclusions The carrying rates of virulence genes of Vibrio parahaemolyticus isolated from patients with diarrhea in Ningbo are high. The drug susceptibility rates are high to common antibiotics,and it should also pay attention to monitoring drug resistance rate and guiding rational antibiotic use in clinic.

Drug resistance and molecular epidemiology of Staphylococcus aureus with suppurative infection

ZHOU Shan, LIU Jiayun, MA Yueyun, ZHOU Ke, ZHOU Lei, HAO Xiaoke, XU Xiuli

2019, 34(2):  140-143.  DOI: 10.3969/j.issn.1673-8640.2019.02.010

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Objective To analyze the drug resistance and molecular epidemiology of Staphylococcus aureus with suppurative infection in the first half of 2017,and to provide a reference for the rational use of antibiotics. Methods The clinical isolates of Staphylococcus aureus with suppurative infection were collected from January to June,2017. Identification and drug susceptibility test were performed by Vitek 2 Compact automatic microbial identification system. The molecular epidemiology of Staphylococcus aureus was analyzed by spa typing and multilocus sequence typing（MLST）. Results A total of 171 isolates ofStaphylococcus aureus were isolated. There were 113 isolates from community infection and 58 isolates from nosocomial infection. The methicillin-resistant Staphylococcus aureus（MRSA） isolation rate in nosocomial infection group was 62.07%,which was higher than that in community infection group（30.97%）. The results of molecular typing showed that the main types in nosocomial and community infection groups were ST239-t030 and ST239-t037,respectively. Conclusions At Xijing Hospital in the first half of 2017,there are differences for Staphylococcus aureus between nosocomial and community infection groups. MRSA clones are relatively concentrated,and nosocomial infection group has spread phenomenon. The control of nosocomial infection should be strengthened,in order to reduce the incidence of nosocomial infection.

Analysis of HPV infection and gene subtypes in physical examination women in Fuzhou

YANG Yang, CHEN Liangyuan, CAO Pengju, CHEN Shaoting, ZHANG Qiuqin, HUANG Chunli, WU Qiumei, CHEN Falin

2019, 34(2):  144-147.  DOI: 10.3969/j.issn.1673-8640.2019.02.0011

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Objective To understand the infection and gene subtype distribution of human papillomavirus（HPV） women in Fuzhou,and to provide a reference for the prevention,diagnosis and treatment of cervical cancer in this area. Methods From February 2014 to December 2017,18 542 women from Fujian Provincial Hospital were enrolled. Universal primer multiplex polymerase chain reaction（PCR） and flow fluorescence hybridization were used to determine 27 HPV gene subtypes in female cervical exfoliated cell samples. Results There were 2 902 cases of HPV positive in 18 542 physical examination women,the positive rate was 15.7%,and 27 gene subtypes were determined. The top 4 high-risk gene subtypes were 52,58,53 and 16,and the top 4 low-risk gene subtypes were 61,81,44 and 55. Totally,2 267 cases（78.1%） were infected with a single type,and 493 cases（17.0%） were double-infected. HPV positive rates of different age groups had statistical significance（P<0.05）. Conclusions The high-risk gene subtypes of HPV infection in physical examination women in Fuzhou are 52,58,53 and 16. The single infection and double infection are the main types of infection. HPV screening over 50-year-old women should be strengthened,which would be of significance for the early detection,prevention and treatment of cervical cancer.

Surveillance and drug resistance of Gram-positive bacteria in Cangnan County

YANG Shaoyou, HUANG Mengqi, ZENG Songfang, LIN Yuanshuang, FANG Guanghui

2019, 34(2):  148-151.  DOI: 10.3969/j.issn.1673-8640.2019.02.012

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Objective To investigate the distribution and drug resistance of Gram-positive bacteria in Cangnan County,and to provide a reference for the clinical prevention and control of nosocomial infection. Methods A total of 517 isolates of Gram-positive bacteria from Cangnan County Second People's Hospital,463 isolates of Gram-positive bacteria from Cangnan County Third People's Hospital and 869 isolates of Gram-positive bacteria from Cangnan County People's Hospital were collected from January 2016 to December 2017. Bacterial identification and drug susceptibility test in vitro were performed. Results Among the 1 849 isolates of Gram-positive bacteria,there were 718 isolates of Staphylococcus aureus（38.83%）,431 isolates of Staphylococcus epidermidis（23.31%）,240 isolates of Staphylococcus haemolyticus（12.98%）,60 isolates of Streptococcus pneumoniae（3.24%）,100 isolates of Streptococcus agalactiae（5.41%）,200 isolates of Enterococcus faecalis（10.82% and 100 isolates of Enterococcus faecium（5.41%）. Among the 718 isolates of Staphylococcus aureus,481 isolates（66.99%） were methicillin-resistant Staphylococcus aureus（MRSA）. All isolates of Staphylococcus were sensitive to tigecycline and linezolid. The isolates ofEnterococcus faecalis were sensitive to vancomycin and tigecycline（100.0%）,and they had different degrees of resistance to linezolid（2.0%）. The isolates of Enterococcus faecium had different degrees of resistance to linezolid（1.0%） and tigecycline（1.0%）. The isolates of Streptococcus pneumoniae were sensitive to tigecycline and vancomycin（100.0%）. Conclusions Among the isolates of Gram-positive bacteria in Cangnan County,Staphylococcus aureus is the main one,followed by Staphylococcus epidermidis,and MRSA is as high as 66.99%. Enterococcus has different degrees of drug resistance.

Performance evaluation of 17 fecal immunochemical tests

LI Pei, ZHU Peichao, SONG Rongwei, TAO Sha

2019, 34(2):  152-158.  DOI: 10.3969/j.issn.1673-8640.2019.02.013

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Objective To investigate the reasons for high false positive rate of fecal immunochemical tests （FIT） through evaluating the performance of 17 common FIT. Methods Simulated feces samples with 14 different levels were prepared. Totally,7 laboratory assistants tested these samples by 15 qualitative FIT（No.1-No.15）and 2 quantitative FIT（No.16 and No.17）. The results were recorded. A total of 2 automatic FIT analyzers,Luminex200 system and OC-Sensor,were used as well. Results A total of 1 666 samples were tested. The positive rate of 11 qualitative FIT was higher than theoretical positive rate（P<0.005）. There was no statistical significance between the positive rate of 2 quantitative FIT and theoretical positive rate（P₁₆=0.43,P₁₇=0.21）. There was a correlation between theoretical concentration and the hemoglobin levels tested by the 2 quantitative FIT（r₁₆=0.88,P<0.000 1;r₁₇=0.92,P<0.000 1）. The sensitivities of qualitative FIT reached 100.00%,and the Kappa values of 11 qualitative FIT were <0.40. The sensitivities of 2 quantitative FIT（No.16 and No.17） were 86.21%,and the specificities were 88.24% and 97.06%,respectively,and the Kappa values were 0.73 and 0.80,respectively.Conclusions The qualitative FIT are cheap and convenient,but the quantity of feces samples and diluents are not standardized well,and the actual cut-off values are lower than given cut-off values,which can lead to the low specificity of qualitative FIT. The cost of quantitative FIT was high,they are not affected by subjective judgment,and the specificity is relatively high.

Immature platelet fraction determination by Sysmex XN automated hematology analyzer for thrombocytopenia

LIN Xiaoyi, CHEN Liting, SHEN Yun

2019, 34(2):  159-161.  DOI: 10.3969/j.issn.1673-8640.2019.02.014

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Objective To evaluate the role of immature platelet fraction（IPF%） determination by Sysmex XN automated hematology analyzer for thrombocytopenia. Methods The platelet（PLT） counts and IPF% of 200 patients with thrombocytopenia [45 patients with primary immune thrombocytopenia（ITP）,40 patients with aplastic anemia（AA）,60 patients undergoing chemotherapy（CT） and 55 patients with chronic liver diseases（CLD）] and 50 healthy subjects（healthy control group） were determined by Sysmex XN automated hematology analyzer. The difference of IPF% was analyzed by GraphPad Prism 6 software,and receiver operating characteristic（ROC） curve analysis was utilized to demonstrate the diagnostic efficiency of IPF% in the assessment of thrombocytopenia. Results The PLT counts of ITP,AA,CT and CLD groups were lower than that of healthy control group（P<0.05）. However,there was no statistical significance between ITP and AA groups and between CT and CLD groups（P>0.05）. IPF% of ITP group was higher than those of healthy control,AA,CT and CLD groups（P<0.05）,while there was no statistical significance between AA,CT and CLD groups and healthy control group（P>0.05）. When the cut-off value of IPF% was 5.8%,the sensitivity was 82.2%,the specificity was 86.8%,and the area under curve（AUC） was 0.92. Conclusions IPF% determination by Sysmex XN automated hematology analyzer plays a role for discriminating hypo- from hyper-proliferative thrombocytopenias.

Gene chip technology in the identification and drug resistance of Mycobacterium

LI Dan, DU Debing, WU Wenyan, WANG Decheng

2019, 34(2):  165-168.  DOI: 10.3969/j.issn.1673-8640.2019.02.016

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Objective To study the role of gene chip technology in the identification and drug resistance of Mycobacterium. Methods The sputum specimens of 680 patients diagnosed as tuberculosis（positive acid fast stain） were collected from January 2014 to December 2016 in the Third People's Hospital of Yichang. Mycobacterium was identified by gene chip technology and culturing. The drug resistance of Mycobacterium tuberculosis（MTB） to rifampin and isoniazid was determined by gene chip technology and culturing. Culturing was used as reference method,and the consistency rate was evaluated. The drug susceptibility test was used as a gold standard for calculating the sensitivity,specificity and consistency rate of gene chip technology. Results A total of 31 isolates of nontuberculous Mycobacterium（NTM） were determined by gene chip technology,and 32 isolates of NTM were determined by culturing. The consistency rate was 96.88%. Compared with drug susceptibility test,the sensitivity of gene chip technology for MTB to rifampin was 89.81%,the specificity was 93.55%,the consistency rate was 92.80%,and the Kappa value was 0.82（P>0.05）. The sensitivity of gene chip technology for MTB to isoniazid was 83.06%,the specificity was 91.39%,the consistency rate was 89.48%,and the Kappa value was 0.73（P>0.05）. Conclusions Gene chip technology is an effective method for accurately screening NTM. Gene chip technology and culturing have high sensitivity and specificity. There is a good consistency between gene chip technology and culturing and drug susceptibility test. Gene chip technology is effective,rapid and safe.

Comparison of phenotypic methods for β-lactamase detection in Staphylococcus aureus

LU Hanwen, ZHOU Wanqing, ZHANG Zhifeng, SHEN Han

2019, 34(2):  169-172.  DOI: 10.3969/j.issn.1673-8640.2019.02.017

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Objective To compare the phenotypic methods for β-lactamase detection in Staphylococcus aureus. Methods A total of 38 isolates with penicillin minimum inhibitory concentration（MIC）≤0.12 μg/mL and 15 isolates with penicillin MIC≥0.25 μg/mL were collected. Penicillin zone edge test（P1 and P10）,nitrocefin test and clover test were used for β-lactamase detection. Polymerase reaction chain（PCR） was used for amplifying blaZ gene. The gene detection method was used as standard,and the difference among these methods were evaluated. Results Among the 38 isolates with MIC≤0.12 μg/mL,by penicillin zone edge test（P10）,2 isolates produced 27 mm diameter of inhibition zone. Totally,15 isolates were positive for blaZ gene,including 2 isolates with MIC=0.12 μg/mL and 13 isolates with MIC≥0.25 μg/mL. Among the 15 isolates,the penicillin zone edge test showed all sensitive（100%）. The nitrocefin test and clover test detected 14 isolates,and the sensitivity was 93.3%. There were 38 isolates being negative for blaZ gene,the nitrocefin test and penicillin zone edge test detected 2 isolates,and the specificity was 94.7%,while clover test detected 3 isolates,and the specificity was 92.1%.Conclusions The phenotypic methods mentioned above are specific and sensitive,which can be used according to clinical needs.

Inhibition of antibacterial peptide from embryonic cells of Musca domestica on human myeloid leukemia cell K562

ZHANG Jinhua, HE Lifang, DENG Renbing, WAN Qihui

2019, 34(2):  173-175.  DOI: 10.3969/j.issn.1673-8640.2019.02.018

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Objective To study the inhibition of antibacterial peptide from embryonic cells of Musca domestica and homoharringtonine（HHT） on human myeloid leukemia cell K562. Methods The embryonic cells of Musca domestica were induced using lipopolysaccharide. Antibacterial peptide was extracted and prepared 80,160,320 and 640 μg/mL level groups by phosphate-buffered saline（PBS）. The medium only adding PBS was used as negative control group. The cell cycle and the cell apoptosis ratio of K562 were determined by flow cytometry. The K562 and human umbilical vein vascular endothelial cells during logarithmic growth phase were prepared as HTT and antibacterial peptide groups. K562 and human umbilical vein vascular endothelial cells without treatment were used as normal control group. The effective inhibition ratios of HHT and antibacterial peptide on K562 and human umbilical vein vascular endothelial cells were determined by flow cytometry. Results Compared with negative control group,the G₀/G₁ of K562 was increased,the S was decreased,and the proliferation index was decreased（P<0.05）. Compared with negative control group,the cell apoptosis ratio of K562 in various level groups had no statistical significance（P>0.05）. The effective inhibition ratios of HHT group and antibacterial peptide group on K562 were increased and had statistical significance compared with normal control group（P<0.05）. The effective inhibition ratio in HHT group on human umbilical vein vascular endothelial cells was higher than those in normal control and antibacterial peptide groups（P<0.05）. Conclusions Antibacterial peptide from embryonic cells of Musca domestica can inhibit tumor cells effectively,but no damage to normal cells.

Development and application of external quality assessment research material for YMDD mutation detection of hepatitis B virus

JIANG Lingli, XIAO Yanqun, WANG Xueliang, BAO Yun, YANG Yixiao, WANG Hualiang

2019, 34(2):  176-179.  DOI: 10.3969/j.issn.1673-8640.2019.02.019

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Objective To prepare hepatitis B virus（HBV） YMDD mutation detection research material,which will be used for evaluating the performance of HBV YMDD mutation detection in Shanghai laboratories by external quality assessment（EQA）. Methods The candidate samples of HBV DNA >5×10⁴ IU/mL with different HBV YMDD mutations were diluted. Real-time fluorescence polymerase chain reaction（PCR）,ligase detection reaction（LDR） and sequencing were used,the homogeneity and stability were evaluated. The sample panel of 2017 2-time EQA program contained 5 samples. The participating laboratories were asked to report their results before deadline. The results from the participants were summarized and analyzed. Results A total of 12 and 10 valid results were received in 2017 2-time EQA program,respectively. There were 66.67%（8/12） laboratories submitted correct results for all samples. The overall consistency rates of HBV YIDD,YVDD,YIDD+YVDD and YMDD samples were 90.63%,86.36%,77.27% and 88.24%,respectively. Conclusions The samples are homogeneous and stable. The results of 2017 EQA program suggest that it is necessary for laboratories to improve the operation for HBV YMDD mutation detection. Quality control in clinical laboratories is essential to assure the accuracy of results.

Retrospective analysis of clinical and laboratory examinations' characteristics of mast cell leukemia

SUN Ye, GUO Ping, XIONG Shumin, WU Rong, KANG Xiangdong

2019, 34(2):  180-184.  DOI: 10.3969/j.issn.1673-8640.2019.02.020

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Objective To analyze the clinical and laboratory examinations' characteristics of a case of mast cell leukemia（MCL） retrospectively,and to understand the clinical and laboratory examinations' characteristics of MCL. Methods Through the clinical and pathologic findings,including complete blood count,bone marrow aspiration,flow cytometry,genetic analysis and DNA sequencing in a case of MCL,the clinical and laboratory examinations' characteristics were analyzed combined with literatures. Results Physical examination revealed recurrent diarrhea,asthenia,severe weight loss,urticaria and marked splenomegaly. In peripheral blood and bone marrow,mast cells（MC）accounted for 42% and 79%,respectively. The immunophenotypes of MC were CD45+,CD117++,CD33++,CD13+,CD11b-,CD2++,CD9++,CD123-,partial CD25+,CD34-,CD38-,HLADR-,CD64- and CD15-. No obvious abnormality was found in genetic analysis. F522C was detected by mutational analysis of KIT gene. Eventually,the case was diagnosed as MCL. Conclusions MCL is a rare hematological malignancy with unique clinical and laboratory examinations' characteristics. It should pay attention to the cell morphology to avoid the misidentification of similar cells,and the molecular biological examination should be emphasized to guide the treatment.