Table of Content

30 June 2018, Volume 33 Issue 6

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XN-20 WPC for the determinations of blast cells and abnormal lymphocytes in peripheral blood

XIA Yonghui, HAN Qingqing, WANG Shoulei, LI Yong

2018, 33(6):  469-471.  DOI: 10.3969/j.issn.1673-8640.2018.06.001

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Objective To investigate the role of XN-20 hematology analyzer white cell precursor channel（WPC） for the determinations of blast cells and abnormal lymphocytes in peripheral blood. Methods By XN-20 WPC,the determinations of blast cells and abnormal lymphocytes in peripheral blood were performed for specimens with Blasts and AbnLympho alarms. The results were compared with those by manual microscopy. The consistency between XN-20 WPC and manual microscopy was evaluated. The repeatabilities and carry-over rates were evaluated as well. Results Compared with manual microscopy,the consistency rates of XN-20 WPC for specimens with Blasts/AbnLympho,Blasts and AbnLympho alarms were 83.3%,42.2% and 42.9%,respectively. The within-run precisions [coefficients of variation（CV）] of 3 specimens with Blasts and AbnLympho alarms for different levels（low,middle and high levels） were 0.36%,1.47%,0.92% and 1.02%,1.49%,0.73%,respectively. The carry-over rates were 0.02%. Conclusions The repeatability of XN-20 WPC is good,and the carry-over rates are low. XN-20 WPC can be used for the analysis of blast cells and abnormal lymphocytes in peripheral blood,which should be confirmed by manual microscopy further.

Relationship of the ultrastructure of acid phosphatase and the apoptosis of spermatogenic cells in infertile patients

WANG Ping, DAI Yanping, YANG Wei, YUE Yingquan, GAO Xiaoqin

2018, 33(6):  472-475.  DOI: 10.3969/j.issn.1673-8640.2018.06.002

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Objective To study the relationship of the ultrastructure of acid phosphatase（ACP） and the apoptosis of spermatogenic cells in infertile patients,and to provide a reference for the diagnosis and treatment of male infertility. Methods Semen specimens were collected from 10 fertile males and 10 infertile male patients,and the semen specimens were washed,centrifugalized,hatched with Gromori's method and embedded in epoxy resin. The ultra-thin sections were made,and the changes of the ultrastructure of ACP and spermatogenic cell apoptosis were determined by histochemistry using enzyme electron microscope. Results The results of histochemistry using enzyme electron microscope showed that the sperm morphology was normal in fertility group,the structures of acrosome,plasma membrane and mitochondria were integrity,and chromatin was regularly configurated. There were immature spermatogenic cells,macrophage cells and neutrophil cells in infertility group,lead-contacted ACP were showed in lysosome,and various abnormal types of acrosome in apoptotic spermatozoa were observed. The apoptotic bodies formed when apoptotic spermatozoa were phagocytosed,and myelin-like patterns formed as well. The concentration of chromatin and electron density in the nucleus of apoptotic spermatogenic cells were high,the nuclear membrane were expanded,ruptured and disintegrated,gap became wider,mitochondria crest were degraded,falled off or disappeared,and the form of apoptotic bodies and the cytoplasmic residual bodies were found. Compared with fertility group,the ACP positive reaction rate of infertility group was higher with statistical significance（P<0.05）. Conclusions The number of apoptotic spermatogenic cells is related closely to the activity and number of ACP. Apoptotic spermatogenic cells are abnormal for infertile males. Histochemistry using enzyme electron microscope is expected to become one way on diagnosing and treating male infertility.

Polymorphism and risk factors of MTHFR C677T for H type hypertension in Kunming Han population

QIAN Jing, SHI Qian, ZHAI Xiuwei, ZHAO Xiaoli, SHAO Jianchun, HU Dachun

2018, 33(6):  476-480.  DOI: 10.3969/j.issn.1673-8640.2018.06.003

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Objective To investigate the correlations of hypertension,serum homocysteine（Hcy） and methylenetetrahydrofolate reductase（MTHFR） C677T polymorphism with H type hypertension in Kunming Han population,and to provide basic information on the prevention and individualized treatment of H type hypertension. Methods A total of 111 patients with H type hypertension were enrolled,and 112 hypertension patients without H type hypertension were enrolled as non-H type hypertension group. Totally,119 healthy subjects were enrolled as healthy control group. Allele specific-polymerase chain reaction（AS-PCR） was used to determine MTHFR C677T polymorphism. Serum Hcy level was determined as well. The risk factors for H type hypertension were analyzed by Logistic regression analysis. Results MTHFR C677T CC,CT and TT genotype frequencies were 49.6%,36.1% and 14.3% in healthy control group. CC,CT and TT genotype frequencies were 24.3%,50.5% and 25.2% in H type hypertension group,and those were 45.5%,35.7% and 18.8% in non-H type hypertension group. The distribution frequencies of CT and TT genotypes in H type hypertension group were higher than those in non-H type hypertension group and healthy control group（P<0.05）. Logistic regression analysis showed that age,MTHFR C677T mutation,the family history of hypertension and drinking were risk factors for H type hypertension,the risk of H type hypertension caused by MTHFR C677T mutation was 2.701 times than those of control groups,and the risk of MTHFR C677T mutation was higher than those of other factors. Conclusions MTHFR C677T allele and polymorphism distribution frequency have regional characteristics among healthy subjects and hypertension patients in Kunming Han population. MTHFR C677T mutation may play an important role in the mechanism of H type hypertension for Kunming Han population.

Correlation of ERAP-1 gene polymorphisms with ankylosing spondylitis in Chinese：a Meta analysis

LI Linyun, PENG Changhua, MEI Bing, DONG Li

2018, 33(6):  481-485.  DOI: 10.3969/j.issn.1673-8640.2018.06.004

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Objective To investigate the correlation of endoplasmic reticulum aminopeptidase 1（ERAP-1）gene single nucleotide polymorphisms（SNP） with ankylosing spondylitis（AS） in Chinese by a Meta analysis. Methods Literatures published from 2009 and 2016 were searched. The frequencies of ERAP-1 gene SNP alleles and genotypes were obtained from original data. The heterogeneities were analyzed by q test. The pooled odds ratios（OR） and 95% confidence intervals（CI） were calculated. Results A total of 9 independent studies were included,and 3 802 Chinese AS cases and 4 930 control cases were enrolled. A total of 10 ERAP-1 gene SNP,including rs27980,rs27037,rs7711564,rs27434,rs27582,rs27044,rs30187,rs10050860,rs2287987 and rs7711564,were analyzed. The Meta analysis showed that rs27037T was a risk factor for AS in Chinese [OR（95%CI） of allele G to T was 1.22（1.12-1.33）,P<0.000 01;OR（95%CI） of genotype GG to TT+GT was 1.34（1.18-1.53）,P<0.000 01],and rs30187C was a protective factor for AS in Chinese [OR（95%CI） of allele T to C was 0.83（0.68-1.00）,P=0.05;OR（95%CI）of genotype TT to CC+TC was 0.80（0.67-0.95）,P=0.01]. Other 9 SNP had no correlation with AS in Chinese（P>0.05）. Conclusions ERAP-1 gene polymorphisms are correlated with AS in Chinese.

Interferon gamma polymorphisms and the susceptibility to nasopharyngeal carcinoma

XIE Qingqing, OUYANG Bohui, LI En, BI Zhongping, ZHU Shengbo, TANG Jian, SUN Yifan

2018, 33(6):  486-490.  DOI: 10.3969/j.issn.1673-8640.2018.06.005

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Objective To investigate the association between interferon gamma（IFN-γ）polymorphisms [rs2430561（+874T/A） and rs1861494（+2109A/G）] and the susceptibility to nasopharyngeal carcinoma（NPC）. Methods A total of 150 NPC patients and 150 healthy subjects were enrolled. IFN-γ +874T/A and +2109A/G were determined by polymerase chain reaction（PCR）-sequence-specific primer（SSP） and PCR-restriction fragment length polymorphism（RFLP）,respectively. Results Compared with AA genotype,the risk of rs2430561 +874T/A TT genotype was decreased [odds ratio（OR）=0.132,95% confidence interval（CI） 0.022-0.800,P=0.028]. However,there was no statistical significance for rs1861494 +2109A/G（P>0.05）. Conclusions IFN-γ rs2430561 +874T/A polymorphisms are associated with the susceptibility to NPC.

Additive effect of serum 25-hydroxyvitamin D₃ in the treatment of depression

LUO Qingxin, CHEN Kaiting, LAN Yanling, MO Xin, CHEN Jiaqiang

2018, 33(6):  491-494.  DOI: 10.3969/j.issn.1673-8640.2018.06.006

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Objective To investigate the relationship of serum 25-hydroxyvitamin D₃ [25（OH）D₃] in the treatment of depression and Hamilton Rating Scale for Depression（HAMD） score,and to evaluate the role of serum 25（OH）D₃ in the treatment of depression. Methods A total of 108 patients with depression were randomly classified into observation group（54 cases） and control group（54 cases）. All patients were given antidepressant Pa Rossi Dean regularly,and the observation group was given vitamin D₃ as well. The analysis of HAMD score was performed for all the patients before treatment and after treatment for 1,3,5 and 7 weeks. The levels of serum 25（OH）D₃ in the 2 groups were determined. Results After treatment for 3,5 and 7 weeks,the levels of serum 25（OH）D₃ in observation group were higher than that in control group（P<0.05）. After treatment for 5 and 7 weeks,HAMD scores in observation group were lower than that in control group（P<0.05）. HAMD score and serum 25（OH）D₃ in observation group had a negative correlation（r=-0.97,P<0.01）. Conclusions For patients with depression,taking vitamin D on the basis of conventional antidepressants can improve the effectiveness of depression treatment. Serum 25（OH）D₃ can be used as a potential indicator of depression efficacy monitoring.

Serum angiotensin-converting enzyme in the early diagnosis of pregnancy-induced hypertension

LI Zhifeng, LIU Dabiao, GUO Lanfang

2018, 33(6):  495-498.  DOI: 10.3969/j.issn.1673-8640.2018.06.007

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Objective To evaluate the role of serum angiotensin-converting enzyme（ACE） in the early diagnosis of pregnancy-induced hypertension. Methods A total of 55 pregnant women with pregnancy-induced hypertension and 45 healthy pregnant women（control group） were enrolled. Renal function indices [creatinine（Cr）,blood urea nitrogen（BUN）,uric acid（UA）,cystatin C（Cys C）,retinol-binding protein（RBP） and beta₂-microglobulin（β₂-MG） levels],glucose（Glu）and ACE level were determined at the middle- and later-periods of pregnancy. Receiver operating characteristic（ROC） curve was used to evaluate the roles of renal function indices and ACE level for the diagnosis of pregnancy-induced hypertension. Results The levels of Cr,BUN,UA,Cys C and RBP in pregnancy-induced hypertension group at the middle-period of pregnancy were higher than those in control group（P<0.05）. The levels of Cr,UA,Glu,β₂-MG,RBP and ACE at the later-period of pregnancy were higher than those in control group（P<0.05）. The levels of BUN,β₂-MG,RBP and ACE in pregnancy-induced hypertension group at the later-period of pregnancy were higher than those at the middle-period of pregnancy（P<0.05）,and the other indices had no statistical significance（P>0.05）. ROC curve showed that the area under curve（AUC） of Cr（0.716） was the biggest at the middle-period of pregnancy. At the later-period of pregnancy,the AUC of ACE（0.924） was the biggest,followed by the AUC of β₂-MG（0.897）. From the middle-period of pregnancy to later-period of pregnancy,ACE had the most obvious increasing of AUC. The sensitivities of β₂-MG for the diagnosis of pregnancy-induced hypertension were increased from 63.6% at the middle-period of pregnancy to 67.3% at the later-period of pregnancy,and the specificities were increased from 55.6% to 88.9%. The sensitivities of ACE were increased from 70.9% to 83.6%,and the specificities were increased from 95.6% to 97.8 %. There was no correlation between β₂-MG and ACE（r=0.278,P>0.05）. Conclusions ACE is a sensitive index for the early diagnosis of pregnancy-induced hypertension.

SAA and hs-CRP for the early diagnosis of infectious diseases in children

TANG Jun, LIN Jianbo

2018, 33(6):  499-502.  DOI: 10.3969/j.issn.1673-8640.2018.06.008

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Objective To investigate the roles of serum amyloid-A（SAA） and high-sensitivity C-reactive protein（hs-CRP） for the early diagnosis of infectious diseases in children. Methods A total of 184 inpatient children were enrolled. Totally,80 cases were confirmed as viral infection（viral infection group）,and 104 cases were diagnosed as bacterial infection（bacterial infection group）. A total of 120 children were enrolled as control group. SAA and hs-CRP levels and white blood cell（WBC） count were determined,and the diagnostic efficiency for infectious diseases in children was analyzed. Results Serum hs-CRP level in bacterial infection group [11.50（6.25,15.00）mg/L] was higher than those in viral infection group [7.00（5.25,17.50）mg/L] and control group [2.25（1.25,4.00）mg/L]. The level of SAA in viral infection group [39.50（22.50,58.00）mg/L] was higher than those in bacterial infection group [24.50（14.25,26.50）mg/L] and control group [4.00（1.50,6.50）mg/L]（P<0.05）. The maximum diagnostic efficiency was obtained,when serum hs-CRP cut-off value was 10 mg/L,with a sensitivity of 76.25%,a specificity of 56.73%,a Youden index of 0.330 and an area under receiver operating characteristic（ROC） curve of 0.52. The maximum diagnostic efficiency was obtained,when SAA cut-off value was 22.5 mg/L,with a sensitivity of 83.75%,a specificity of 66.35%,a Youden index of 0.501 and an area under ROC curve of 0.64. Conclusions SAA level has better diagnostic efficiency than serum hs-CRP level for the early diagnosis of infectious diseases in children,which deserves clinical application and promotion.

Serum IgG4 and IgE in screening food intolerance in children with allergic dermatosis

LUO Xingxing, PI Xiaobing, ZHU Changlin, LI Weixuan

2018, 33(6):  503-507.  DOI: 10.3969/j.issn.1673-8640.2018.06.009

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Objective To investigate the role of serum IgG4 and IgE in screening food intolerance in children with allergic dermatosis under 12 years old. Methods Total IgG4,total IgE,food intolerance specific IgG4（sIgG4） and specific IgE（sIgE） in 165 children with allergic dermatosis（patient group） and 88 healthy children（control group） were determined. The difference was compared among different types of allergic dermatosis,different ages and different onset times. Results The levels of serum total IgG4 and total IgE in patient group were higher than those in control group（P<0.05）. Serum total IgG4 and total IgE levels with different types of allergic dermatosis were higher than those in control group（P<0.05）,but there was no statistical significance for different types of allergic dermatosis（P>0.05）. The positive rate of serum sIgG4 in children with digestive symptoms（66.2%） was higher than that in children without digestive symptoms（47.7%,P=0.017）,but there was no statistical significance for sIgE positive rate between the 2 groups（35.1% and 36.4%,P=0.862）. The positive rate of serum sIgG4 in patient group（58.2%） was higher than that of serum sIgE（35.8%,P<0.01）. The positive rate of serum sIgG4 and sIgE were increased with age. The positive rate of sIgG4 in children with different ages was higher than that of sIgE,and there was statistical significance between 1-3 years old and 7-12 years old children（P=0.031,P=0.008）. The positive rates of sIgG4 for egg,mango/pineapple/peach,shrimp/crab/shell and fish were high. The positive rate of serum sIgE was higher than that of serum sIgG4 for onset <2 h（P<0.01）,which was lower than that of serum sIgG4 for onset ≥2 h（P<0.01）. Conclusions The positive rates of serum sIgG4 and sIgE increase with age. The determination of sIgG4 is prefer for children with chronic allergic dermatosis,and the determination of sIgE in children with acute onset is more valuable.

WFDC2 encoding HE4 in the diagnosis and curative effect monitoring of pulmonary carcinoma

LUO Jialing, PENG Xia, LIN Kun, LI Li

2018, 33(6):  508-511.  DOI: 10.3969/j.issn.1673-8640.2018.06.010

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Objective To investigate the role of WFDC2 encoding human epididymis protein 4（HE4） in the early diagnosis and curative effect monitoring of pulmonary carcinoma. Methods A total of 226 pulmonary carcinoma patients（pulmonary carcinoma group）,128 patients with benign pulmonary disease（benign disease control group） and 100 healthy subjects（healthy control group） were enrolled. WFDC2 encoding HE4 was determined by enzyme immunoassay. The levels of carcinoembryonic antigen（CEA）,cytokeratin 19 fragment（CYFRA21-1）,neuron-specific enolase（NSE） and squamous cell carcinoma（SCC） related antigen were determined by chemiluminescence. The areas under receiver operating characteristic（ROC）curves（AUC） of HE4,CEA,CYFRA21-1,NSE and SCC related antigen in the 3 groups were evaluated. Results Serum HE4 level of pulmonary carcinoma group was 93.29（61.74,160.28） pmol/L,which was higher than those of benign disease control group [76.47（55.35,129.85） pmol/L] and healthy control group [45.64（36.60,62.35） pmol/L]（P<0.05）. There was no statistical significance for serum HE4 level with different pathological types of pulmonary carcinoma（P>0.05）. In the diagnosis of pulmonary carcinoma,the AUC of HE4 was 0.860,and HE4 had good diagnostic efficiency. The AUC of HE4 combined with CEA,CYFRA21-1,NSE and SCC related antigen was 0.906,which was higher than those of 5 single determinations. There was statistical significance for serum HE4 levels with different clinical stages of pulmonary carcinoma（P<0.05）. Serum HE4 levels were elevated after chemotherapy（P<0.05）. Conclusions WFDC2 encoding HE4 can be used as a marker for the early diagnosis and curative effect monitoring of pulmonary carcinoma.

Tumor markers for the combined screening of female genital tract cancer and breast cancer

WU Yongchun

2018, 33(6):  512-515.  DOI: 10.3969/j.issn.1673-8640.2018.06.011

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Objective To investigate the diagnostic role of the combined determination of serum alpha fetoprotein（AFP）,carcinoembryonic antigen（CEA）,carbohydrate antigen 199（CA199）,carbohydrate antigen 153（CA153）and carbohydrate antigen 125（CA125） in female genital tract cancer and breast cancer. Methods Totally,55 female genital tract cancer and breast cancer patients,55 benign inflammatory patients and 55 healthy subjects were enrolled. Serum levels of AFP,CEA,CA199,CA153 and CA125 were determined,and the results were analyzed statistically. The sensitivities and specificities of the 5 markers for the diagnosis of female genital tract cancer and breast cancer were compared. Results The levels of AFP,CEA,CA199,CA153 and CA125 in cancer group were higher than those in benign inflammatory and healthy control groups（P<0.05）. The sensitivity and specificity of the combined determination of the 5 markers were higher than those of the single determinations. Conclusions The combined determination of the 5 markers has a diagnostic value in the screening of female genital tract cancer and breast cancer,and it is helpful for early screening and early diagnosis.

Single and combined determinations of serum β-HCG ratio,progesterone and endometrial thickness for predicting ectopic pregnancy

XIE Jun, ZHENG Xuan, MA Jing, JIN Qin

2018, 33(6):  516-520.  DOI: 10.3969/j.issn.1673-8640.2018.06.012

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Objective To investigate the roles of single and combined determinations of serum human chorionic gonadotropin（β-HCG） ratio,progesterone and endometrial thickness for predicting ectopic pregnancy. Methods A total of 300 suspected patients with ectopic pregnancy were enrolled,they were proved to be ectopic pregnancy or intrauterine pregnancy later,and their clinical data were analyzed retrospectively. Receiver operating characteristic（ROC） curve and Logistic regression equation were used to analyze the roles of serum β-HCG ratio,progesterone and endometrial thickness determinations for predicting ectopic pregnancy. The predictive cut-off value of each index was determined,and the sensitivity and specificity of each index of single and combined determinations were evaluated. Results Serum β-HCG ratio,progesterone and endometrial thickness in ectopic pregnancy group were lower than those in normal pregnancy group（P<0.001）. The single determinations of serum β-HCG ratio,progesterone and endometrial thickness were valuable for the prediction of ectopic pregnancy. When the cut-off value of progesterone was 55.3 nmol/L,the sensitivity was 90.4%,and the specificity was 78.6%. When the cut-off value of serum β-HCG ratio was 2.01,the sensitivity was 86.5%,and the specificity was 83.9%. When the cut-off value of endometrial thickness was 11.5 mm,the sensitivity was 86.5%,and the specificity was 60.7%. The sensitivity of the combined determination was 94.6%,and the specificity was 88.5%. The areas under the ROC curves of single and combined determinations were 0.852 for the determination of serum β-HCG ratio,0.845 for the determination of progesterone,0.736 for the determination of endometrial thickness,0.887 for the combined determination of progesterone and endometrial thickness,0.868 for the combined determination of serum β-HCG ratio and progesterone,0.867 for the combined determination of serum β-HCG ratio and endometrial thickness and 0.950 for the combined determination of 3 indices. Conclusions The combined determination of serum β-HCG ratio, progesterone and endometrial thickness for predicting ectopic pregnancy has good efficiency than that of any single determination for the early diagnosis of ectopic pregnancy.

Pathogen analysis of 63 isolates of anaerobic bacteria for bloodstream infection and investigation of primary infectious diseases

XIONG Yan, ZHANG Hong, CHEN Yantian

2018, 33(6):  521-524.  DOI: 10.3969/j.issn.1673-8640.2018.06.013

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Objective To study the pathogen distribution and drug resistance of anaerobic bacteria for bloodstream infection,to investigate primary infectious diseases,and to provide a reference on controlling the occurrence of anaerobic bacterium bloodstream infection. Methods BacT/ALERT 3D automatic blood culture system was used to perform blood culturing,and ATB expression automated analysis system was used for bacterial identification and drug susceptibility. Results Among 63 isolates of anaerobic bacteria,Clostridium perfringens accounted for 31.7%,Bacteroides fragili accounted for 25.4%,and Fusobacterium fragiformisaccounted for 11.1%. The drug resistance rates to metronidazole,carbapenem,chloramphenicol and enzyme inhibitors were all <10.0%,and the drug resistance rate to metronidazole was <2.0%. The drug resistance rates to penicillin and clindamycin were >30.0%. Gram-positive anaerobic bacteria were susceptible to cefoxitin and cefotetan. The drug resistance rates of Gram-negative anaerobic bacteria to cefoxitin and cefotetan were 15.4% and 20.5%. The primary infectious diseases were postoperative infection after intestinal surgery（22.2%）,empyema （17.5%） and appendix perforation（11.1%）. Conclusions Gram-negative no-spore forming anaerobic bacteria and Gram-positive Clostridium are main anaerobic bacteria for bloodstream infection. When the results of drug susceptibility test were not available,the clinicians could choose metronidazole,carbapenem and enzyme inhibitors as first choice. The primary infectious diseases mainly occur in intestine,chest and female reproductive tract. It should be prevented as soon as possible,which is conducive to reducing the occurrence and development of bloodstream infection.

Clinical characteristics,isolation rate and drug resistance of Stenotrophomonas maltophilia

LU Taohong, ZHANG Qingfang, ZHU Xiaoli, YUAN Dandan, LI Xin

2018, 33(6):  525-529.  DOI: 10.3969/j.issn.1673-8640.2018.06.014

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Objective To investigate the risk factors,clinical characteristics,isolation rate and drug resistance of Stenotrophomonas maltophilia infection,and to provide a reference for its clinical prevention and treatment. Methods A total of 313 isolates of Stenotrophomonas maltophilia were collected from January 2012 to December 2016,and their data and antibiotic susceptibility results were analyzed retrospectively. Results The 313 isolates of Stenotrophomonas maltophilia were collected from 260 cases of respiratory tract infection and 53 cases of non-respiratory tract infection,and the major distribution was in Department of Respiratory. The high risk factors were old age,usage of antibiotics for long time,primary pulmonary and so on. The isolation rates of Stenotrophomonas maltophilia isolated from 2012 to 2016 in Gram-negative bacilli were 5.3%（97/1 831）,3.2%（59/1 818）,3.7%（60/1 616）,2.3%（51/2 212） and 1.5%（46/3 028）（P<0.05）. The drug resistance rates of 313 isolates of Stenotrophomonas maltophilia against sulfamethoxazole-trimethoprim,levofloxacin,minocycline,ceftazidime,ticarcillin-clavulanic acid and chloramphenicol were 29.6%,20.4%,3.8%,68.9%,44.4% and 20.9%. The drug resistance rate to ceftazidime was increased,and the drug resistance rates to the other drugs were decreased year by year（P<0.001）. The drug resistance to ceftazidime,minocycline and chloramphenicol in isolates from respiratory tract samples had statistical significance compared with those from non-respiratory tract samples（P<0.05）. Conclusions The risk factors of Stenotrophomonas maltophilia infection are old age,primary pulmonary,usage of antibiotics,tracheotomy,intensive care unit admission and so on. Rational antibiotic use is important. Stenotrophomonas maltophilia infection should be treated with minocycline combined with other antibiotics.

Establishment of PCR and ASE-based detection for SNP

SHAN Hongbo, JIN Yanan

2018, 33(6):  530-535.  DOI: 10.3969/j.issn.1673-8640.2018.06.015

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Objective To establish polymerase chain reaction（PCR）,allele-specific extension（ASE） and nucleic acid detection strip-based detection for single nucleotide polymorphisms（SNP）. Methods The specific gene sequence containing SNP sites was amplified by PCR. For each SNP site,ASE primers labeled with A were extended. The ASE reaction products can bind to probe labeled with B,and hybridization products containing A and B simultaneously were formed. The products can be detected by disposable amplicon cross-contamination proof device containing a nucleic acid detection strip,and the detection of SNP genotypes can be accomplished. Results Ten-fold serial dilutions of quantified human genomic DNA were used to determine the sensitivity of PCR-ASE（88 ng/reaction）. The ability of duplex PCR-ASE with the products can be detected by a single nucleic acid detection strip. A total of 19 samples representing 5 common SNP were detected by PCR-ASE,and the results had the consistency of 100% with DNA sequencing. Conclusions PCR-ASE is simple and accurate detection for SNP.

Rapid detection of KPC-2 producing Klebsiella pneumoniae ST11 by matrix-assisted laser desorption ionization time-of-flight mass spectrometry

YU Jiajia, LIU Jingxian, LI Yuanrui, YU Jing, LIU Ying

2018, 33(6):  536-542.  DOI: 10.3969/j.issn.1673-8640.2018.06.016

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Objective To screen and verify the specific peak of KPC-2 producing Klebsiella pneumoniae ST11 by matrix-assisted laser desorption ionization time-of-flight mass spectrometry（MALDI-TOF MS）,and to establish a rapid method for the detection of Klebsiella pneumoniae. Methods A total of 118 isolates of Klebsiella pneumoniae which had been confirmed by carbapenemase gene detection and classified by multilocus sequence typing（MLST） were collected as specific peak screening group,67 isolates of them were KPC-2 producing Klebsiella pneumoniae ST11,and 51 isolates of them were non-KPC-2 producing Klebsiella pneumoniae ST11. Clinpro Tools 3.0 software and Flex analysis 3.0 profile analysis software were used to screen the specific peak. Other 89 isolates of Klebsiella pneumoniae which had detected by carbapenemase gene detection and classified by MLST were selected as specific peak verifying group. Mass spectra were acquired strictly according to experimental conditions,and the specificity and repeatability of specific peak were detected. Finally,95 isolates of Klebsiella pneumoniae and their mass spectra were collected. These isolates were distinguished directly by the specific peak. Meanwhile,carbapenemase gene detection and MLST were performed for these isolates in order to evaluate the efficiency of specific peak. Results The specific peak of KPC-2 producing Klebsiella pneumoniae ST11 was m/z 4 521. In the experimental conditions,the specificity and sensitivity were 98.1% and 97.3%,and the repeatability of specific peak was 97.4%. In the clinical conditions,using mass spectra,the specificity and sensitivity were 95.2% and 96.9%,respectively. Conclusions The specific peak m/z 4 521 on MALDI-TOF MS is the specific peak of KPC-2 producing Klebsiella pneumoniae ST11. KPC-2 producing Klebsiella pneumoniae ST11 can be detected directly according to the specific peak with high specificity and sensitivity,which can provide a reference for clinical treatment and nosocomial infection control rapidly and accurately.

Determination of reverse triiodothyronine by chemiluminescence assay

SONG Yunxiao, WU Jianmin, ZHANG Haichen

2018, 33(6):  543-548.  DOI: 10.3969/j.issn.1673-8640.2018.06.017

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Objective To evaluate the performance of reverse triiodothyronine（rT₃） determination by chemiluminescence assay. Methods The precision,accuracy,analytical sensitivity,analytical specificity,linearity,reference range and methodological comparison of rT₃ determination by chemiluminescence assay were evaluated according to EP documents of the Clinical and Laboratory Standards Institute（CLSI）. The levels of 3,5,3'-triiodothyronine（T₃）,thyroxine（T₄）,thyroid-stimulating hormone（TSH） and rT₃ were determined in 50 healthy subjects（healthy control group） and 131 patients,including 30 preliminarily diagnosed hyperthyroidism patients（hyperthyroidism group）,28 hyperthyroidism patients with treatment for 1 month（hyperthyroidism treatment group）,25 preliminarily diagnosed hypothyroidism patients（hypothyroidism group）,23 hypothyroidism patients with treatment for 1 month（hypothyroidism treatment group） and 25 primary non-thyroid diseases [euthyroid sick syndrome （ESS）]. Results For high-level and low-level quality control materials of rT₃,the within-run coefficients of variation（CV）were 4.82% and 4.95%,and the between-run CV were 7.51% and 5.90%,respectively. The biases were -1% and 1%. The linearity was 0.05-10.07 ng/mL,and the reference range was 0.20-0.95 ng/mL. There was a good correlation between chemiluminescence assay with radioimmunoassay（r=0.952）. Compared with healthy control group,the levels of rT₃,T₃ and T₄ in hyperthyroidism group were increased（P<0.05）,and TSH was decreased（P<0.05）. In hypothyroidism group,the levels of rT₃ and T₄ were decreased（P<0.05）,and TSH was increased（P<0.05）. Compared with hyperthyroidism group,hyperthyroidism treatment group had decreased rT₃,T₃ and T₄ levels（P<0.05）,and TSH was increased（P<0.05）. Compared with hypothyroidism group,hypothyroidism treatment group had increased rT₃ and T₄ levels（P<0.05）,and TSH decreased（P<0.05）. The level of rT₃ in ESS group was higher than those in hypothyroidism and healthy control groups（P<0.05）,the levels of T₃ and T₄ were lower than those in healthy control group（P<0.05）,there was no statistical significance compared with hypothyroidism group（P>0.05）,TSH level was lower than that in hypothyroidism group（P<0.05）,and there was no statistical significance compared with healthy control group（P>0.05）. Conclusions The determination of rT₃ by chemiluminescence assay has good precision,accuracy,specificity and linearity,which is suitable for large scale sample determination. It can be used for the differential diagnosis of hypothyroidism and ESS.

Correlation between the methylation status of C-erbB-2 gene promoter region CpG island and androgen receptor expression in breast cancer

YAN Peiyi, YUAN Ya, ZHANG Ji, ZHU Qi, ZOU Yuhan, WANG Gaoying, JIN Shu

2018, 33(6):  549-555.  DOI: 10.3969/j.issn.1673-8640.2018.06.018

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Objective To investigate the correlation between androgen receptor（AR） and the methylation status of human epidermal growth factor receptor（C-erbB-2） gene promoter region CpG island in breast cancer,and to study the relationship between AR and clinical prognosis. Methods A total of 76 patients with breast cancer and 55 patients with benign breast tumor were enrolled,and the methylation status of C-erbB-2 gene promoter region CpG island and the expression level of AR were determined by methylation-specific polymerase chain reaction（MSP） and immunohistochemistry（IHC）,respectively. Based on clinical and pathological data,the relationship between AR and the methylation status of C-erbB-2 gene promoter region CpG island was evaluated,and the prognosis analysis was performed. Results The methylation proportion of C-erbB-2 gene promoter region CpG island in AR positive breast cancer tissue [74.58%（44/59）] was higher than that in AR negative breast cancer tissue [47.06%（8/17）]（P=0.032）. The methylation of C-erbB-2 gene promoter region CpG island was positively correlated with AR expression（r=0.247,P=0.032）,and the tumor free survival rate of AR positive patients was higher than that of AR negative patients（P<0.05）. Conclusions The expression status of AR in breast cancer tissue can be used as one of prognosis predictors. The methylation of C-erbB-2 gene promoter region CpG island is positively related to the expression status of AR,which provides a new reference for hormone therapy and breast cancer prognosis.

Influence of baicalein on in vitro anti-tumor activity of docetaxel for prostate cancer

YING Xiao, WANG Yuying, WANG Zaihong, WANG Zhenhua

2018, 33(6):  556-562.  DOI: 10.3969/j.issn.1673-8640.2018.06.019

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Objective To investigate the influence and mechanism of baicalein on in vitro anti-tumor activity of docetaxel for prostate cancer. Methods According to different processing modes,human prostate cancer LNCaP cells were classified into control group（empty plasmid transfection）,baicalein group（10 mmol/L baicalein）,docetaxel group（1 nmol/L docetaxel）,docetaxel+baicalein group（1 nmol/L docetaxel and 10 mmol/L baicalein） and docetaxel+baicalein+murine double minute 2（MDM2） plasmid group （1 nmol/L docetaxel and 10 mmol/L baicalein after MDM2 plasmid transfection）. 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay was performed to determine the activity of LNCaP cells. Flow cytometry was used to determine the apoptosis of LNCaP cells. Western blotting analysis was performed to determine the expression levels of MDM2,protein 53（p53）,phorbol-12-myriistate-13-acetate-induced protein 1（PMAIP1,also known as Noxa） and p53 upregulated modulator of apoptosis（Puma） and the activation levels of cysteine-containing aspartate-specific protease（Caspase）-9 and Caspase-3 in LNCaP cells. Results The activity inhibitory rate and apoptotic rate of LNCaP cells in docetaxel+baicalein group were higher than those in docetaxel+baicalein+MDM2 plasmid group,docetaxel group,baicalein group and control group（P<0.05）. The levels of MDM2 mRNA and MDM2 protein in LNCaP cells of baicalein group and docetaxel+baicalein group were lower than those in docetaxel+baicalein+MDM2 plasmid group,docetaxel group and control group（P<0.05）. The p53,Puma and Noxa expression levels,the releasing levels of cytochrome c and the activation levels of Caspase-9 and Caspase-3 in docetaxel+baicalein group were higher than those in docetaxel+baicalein+MDM2 plasmid group,docetaxel group,baicalein group and control group（P<0.05）,while relatively mitochondrial membrane potential was lower than those in the other groups（P<0.05）. Conclusions Baicalein increases the in vitro anti-tumor activity of docetaxel for prostate cancer through MDM2/p53.

Six sigma for evaluating the performance of blood cell analysis internal quality control inter-laboratory comparison among 17 laboratories from Grade 3 Class A hospitals in Hebei

WANG Huiru, LI Guixia, HUANG Jing, WU Hao, DI Yang

2018, 33(6):  563-566.  DOI: 10.3969/j.issn.1673-8640.2018.06.020

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Objective To use six sigma（6σ） for evaluating the performance of blood cell analysis internal quality control（IQC）inter-laboratory comparison among 17 laboratories from Grade 3 Class A hospitals in Hebei,and to improve the quality of blood cell analysis. Methods The data of IQC inter-laboratory comparison among 17 laboratories from Grade 3 Class A hospitals in Hebei were collected,and the determination items included white blood cell（WBC） count,red blood cell（RBC） count,hemoglobin（Hb） and platelet （PLT） count,and coefficients of variation（CV）,biases（Bias）,sigma（σ） values and quality goal indices（QGI） were calculated. Results Using WS/T 406—2012 as standard for allowable total error（TEa）,the laboratories with the performance of WBC count,RBC count,Hb and PLT count with 2 levels （middle level and high level）≥6σ accounted for 52.94% and 47.06%,17.65% and 17.65%,35.30% and 23.53%,52.94% and 58.82%,and those with 4≤σ<6 accounted for 35.30% and 47.06%,29.41% and 29.41%,47.05% and 47.06%,35.30% and 35.30%,respectively. Conclusions The performance of blood cell analysis IQC inter-laboratory comparison among 17 laboratories is good,and 6σ is a unified and simple method for IQC.

Investigation and analysis on quality indicators for clinical laboratories in Shanghai

ZHU Jun, YANG Xue, LOU Jiao, XU Chong

2018, 33(6):  567-571.  DOI: 10.3969/j.issn.1673-8640.2018.06.021

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Objective To study the status of clinical laboratories in Shanghai,and to provide a reference for establishing management regulation for improving laboratory quality in Shanghai through the analysis on Shanghai clinical laboratories and 15 quality indicators（QI）. Methods The data of Shanghai clinical laboratories and 15 QI were collected by questionnaire survey. The data were processed using Microsoft Excel 2016 software and analyzed using SPSS 20.0 software. A total of 13 QI expressed in rate were evaluated by converting them into sigma（σ）metric,and the results were compared with the corresponding data reflecting the overall level of national laboratories. Results The σ level of inter-laboratory comparison was 3,and the others were all >3. The σ levels of the 13 QI expressed in rate were all higher than those from nationwide at same period. Pre-analytical turn-around time（TAT）in emergency examination were all between 15 and 50 min. TAT in routine examination were all between 70 and 80 min. Intra-laboratory TAT of clinical immunology were the longest,which were 492 min for routine examination and 223 min for emergency examination. Conclusions The status reflected by the implementation rate of internal quality control and the rate of inter-laboratory comparison need to be improved. Laboratories should strengthen the establishment of laboratory information system（LIS） to ensure the convenience and reliability of datum collection.

Research progress in markers for the diagnosis of colorectal cancer

YANG Jing, ZHU Linmin, CHEN Xingguo

2018, 33(6):  572-577.  DOI: 10.3969/j.issn.1673-8640.2018.06.022

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The incidence of colorectal cancer is increasing in China. Colonoscopy is the gold standard for the diagnosis of colorectal cancer. However,it is not easy to be accepted by patients due to pain,discomfort,bleeding,perforation and other complications,since it is an invasive examination. Non-invasive screening of blood and stool markers is more likely to be accepted. The determinations of fecal occult blood and serum carbohydrate antigen（CA） are the most commonly used screening methods. In recent years,tumor type M2 pyruvate kinase（tM2-PK） is valuable in screening. With the development of molecular biology,promoter hypermethylation in epigenetics and aberrant expression of microRNA（miRNA） in the determinations of blood and feces are expected to improve the early determination rate of colorectal cancer. However,a large number of clinical studies are still needed to confirm these methods.