Table of Content

20 January 2017, Volume 32 Issue 1

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Orginal Article

Correlation of circulating insulin-like growth factor-1 and clinicopathological parameters in colorectal cancer

XU Dandan, WANG Ruyao, LIU Rui, ZHAO Lijie, HU Liling, SUN Ao, WU Ping

2017, 32(1):  1-4.  DOI: 10.3969/j.issn.1673-8640.2017.01.001

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Objective To investigate circulating insulin-like growth factor-1（IGF-1） in the progression of colorectal cancer （CRC）. Methods The levels of circulating IGF-1,carcinoembryonic antigen（CEA） and carbohydrate antigen 19-9 （CA19-9） were determined by chemiluminescence in 196 patients with CRC and 241 healthy controls. Results The level of circulating IGF-1 in CRC group [（143.86±55.07）ng/mL] was higher than that in healthy control group [（133.60±46.76）ng/mL]（P<0.05）. In healthy control group,the level of circulating IGF-1 in male group was higher than that in female group,which decreased with the increasing of age（P<0.05）. The risk of CRC was high relatively in subjects with high circulating IGF-1 [odds ratio（OR）=1.72,95% confidence interval（CI） 1.049-2.835,P<0.05]. The level of circulating IGF-1 was higher in CRC group with tumor≥5 cm,at advanced stages（stage Ⅲ and stages Ⅳ）,with low differentiation and with lymphoid node metastasis than those with tumor <5 cm,at early stages （stage Ⅰ and stage Ⅱ）,with middle and high differentiation and without lymphoid node metastasis （P<0.05）. The levels of CEA and CA19-9 were correlated to the pathological staging of tumor. The sensitivity of circulating IGF-1 combined determination with CEA and CA19-9 （58.16%）was higher than those of single items （23.98% for circulating IGF-1,32.22% for CEA and 15.31% for CA19-9）（P<0.05）. Conclusions Circulating IGF-1 is an independent risk factor for CRC and plays an important role in the progression of CRC,and it has clinical significance for the prognosis evaluation of CRC. Circulating IGF-1 combined determination with CEA and CA19-9 could improve the determination sensitivity for CRC.

Combined determination of angiopoietin-2,vascular endothelial growth factor and carcinoembryonic antigen in pleural effusion of patients with lung cancer

ZHANG Li, LIU Hong

2017, 32(1):  5-7.  DOI: 10.3969/j.issn.1673-8640.2017.01.002

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Objective To investigate the levels of angiopoietin-2 （Ang-2）,vascular endothelial growth factor （VEGF） and carcinoembryonic antigen（CEA） in pleural effusion for the diagnosis of lung cancer with pleural effusion and different pathological types of lung cancer. Methods A total of 79 pleural effusion samples were collected from 43 lung cancer patients with pleural effusion（12 cases of squamous carcinoma,20 cases of adenocarcinoma and 11 cases of small-cell lung cancer） and 36 patients with benign pleural effusion. The levels of Ang-2 and VEGF were determined by enzyme-linked immunosorbent assay（ELISA）,and the level of CEA was determined by radioimmunoassay. The levels of different pathological types of lung cancer were compared. Results The levels of Ang-2,VEGF and CEA were higher in lung cancer with pleural effusion group than those in benign pleural effusion group （P<0.05）. The positive rate of CEA in adenocarcinoma group was higher than those in squamous carcinoma and small-cell lung cancer groups （P<0.05）,but the positive rates of Ang-2 and VEGF had no statistical significance （P>0.05）. Conclusions The determinations of Ang-2,VEGF and CEA play roles for the diagnosis of lung cancer with pleural effusion,and the combined determination can improve diagnosis efficiency. CEA is potential for the diagnosis of adenocarcinoma,and there is no correlation of Ang-2 and VEGF with different pathological types of lung cancer.

Causes and disposal scheme for grey zone samples of anti-HCV antibody determination

LU Yinhua, JIANG Lingli, ZHU Yuqing, XU Chong, ZHAO Xiaojun, CAO Danru, ZHU Lingfeng, GU Zhidong

2017, 32(1):  8-13.  DOI: 10.3969/j.issn.1673-8640.2017.01.003

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Objective To investigate the causes and disposal scheme for grey zone samples of anti-hepatitis C virus（HCV） antibody determination. Methods Anti-HCV antibody were determined by 5 domestic kits [4 kits using indirect enzyme-linked immunosorbent assay（ELISA） and 1 kit using double antigen sandwich method] and 1 imported kit using chemiluminescence immunoassay. Samples with inconsistent results were further determined by recombinant immunoblot assay（RIBA）. Then,real-time fluorescence quantitation polymerase chain reaction（PCR） was used to determine indeterminate samples for HCV RNA in RIBA. A total of 12 samples in window period were determined by HCV RNA for copies and genotypes. Results Except kit F,the other 5 kits showed review negative coincidence rates ≥ 90%. Review positive coincidence rates were increased with the increasing of initial S/CO ratio. When S/CO ratio was >15.01,6 kits showed review positive coincidence rates > 95%. The false negative rates of single 1 kit were 27.8%-94.4%. The false negative rates of 2 domestic kits randomly were 8.3%-61.1%,the false negative rates of 2 domestic kits randomly with 1 imported kit were 5.6% -22.2%,and the false negative rates of 2 domestic kits randomly using indirect ELISA with 1 domestic kit using double antigen sandwich method were 2.2%-13.9%. The results of 12 samples in window period by 6 kits were negative or positive partly in grey zone,the results by RIBA were negative or indeterminate,and the results for HCV RNA was weakly positive. The 8 of 12 samples in window period were genotype 1b,and the other 4 samples had no genotype. Conclusions When the results are in grey zone,it should use at least 2 different kits for reviewing. Kit using double antigen sandwich method or imported kit can be introduced for supplement. In order to reduce the missing determination of samples in window period,it suggests that HCV RNA determination can be used as supplement.

Percentages of peripheral blood CD4⁺CD25⁺CD127^low T cells before and after the treatment of paternal lymphocyte induced immunization in patients with unexplained recurrent spontaneous abortion

LI Cui, SHI Youhao, WU Jiangnan, YING Chunmei

2017, 32(1):  14-17.  DOI: 10.3969/j.issn.1673-8640.2017.01.004

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Objective To investigate the expression of peripheral blood CD4⁺CD25⁺CD127^1ow T cells in regulatory T cells （Treg） of patients with unexplained recurrent spontaneous abortion（URSA）,to analyze the percentages before and after the treatment of paternal lymphocyte induced immunization,and to evaluate its probability as a diagnosis indicator for abortion. Methods By flow cytometry,the percentages of peripheral blood CD4⁺CD25⁺CD127^1ow T cells were determined in 73 patients with URSA before the treatment,76 healthy women with early pregnancy and 69 healthy women without pregnancy. A total of 51 patients with URSA underwent the treatment of paternal lymphocyte induced immunization for 2-3 treatment courses. Their percentages were determined as well. Results Before the treatment,the percentage of CD4⁺CD25⁺CD127^1ow T cells in URSA group （2.73%±0.56%） was lower than those in healthy pregnancy and non-pregnancy groups（4.36%±1.07% and 2.93%±0.60%）（P<0.05）. After the treatment,the percentage（3.72%±1.15%）was higher than that before the treatment（P<0.05）. The pregnancy outcome of 42 patients after the treatment was good,while the treatment for the other 9 patients failed. Conclusions The percentage of peripheral blood CD4⁺CD25⁺CD127^1ow T cells in URSA patients is lower than those in healthy pregnant and non-pregnant women,which can be as a diagnosis indicator for URSA,and it can be improved by the treatment of paternal lymphocyte induced immunization.

Pathogenic characteristics of hand-foot-mouth disease in Tangxian of Baoding from 2012 to 2015

WANG Yuxiao, ZHAO Can, LI Juanjuan, DI Keping, TIAN Hongbiao, WANG Caihong

2017, 32(1):  18-21.  DOI: 10.3969/j.issn.1673-8640.2017.01.005

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Objective To study the pathogenic characteristics of hand-foot-mouth disease in Tangxian of Baoding from 2012 to 2015,and to provide a reference for the prevention and controlling of hand-foot-mouth disease. Methods Fluorescence reverse transcription polymerase chain reaction（PCR） was used to determine throat specimens of 1 652 hand-foot-mouth disease suspected cases for enterovirus,enterovirus 71（EV71） and coxsackievirus A16（CA16）. Results Of the 1 652 hand-foot-mouth disease suspected cases,there were 511 positive cases for enterovirus. The positive cases for EV71,CA16 and other enteroviruses accounted for 31.31%,21.72% and 46.97%,respectively. In 2012,the primary pathogen causing hand-foot-mouth disease was EV71,but other enteroviruses became dominant pathogens for hand-foot-mouth disease from 2013 to 2015. The positive cases were determined all year round and distributed seasonally,with a peak appearing from May to August. Hand-foot-mouth disease changed its pathogenic constitution in different seasons. The main pathogens were other enteroviruses from January to June. The peak of EV71 appeared from July to September. CA16 became dominant pathogen from October to December. A majority of positive cases were children < 5 years old. EV71 was a superior epidemiological enterovirus for severe hand-foot-mouth disease. Conclusions The pathogenic monitoring of hand-foot-mouth disease needs be enhanced. The comprehensive controlling measures should be taken in key population from May to August,in order to prevent form hand-foot-mouth disease outbreak and spreading.

Establishment and verification of microscopy review rules in AVE automatic urine analysis

LIU Wanchao, CHEN Longmei, YANG Zhenhua, LI Dong

2017, 32(1):  22-25.  DOI: 10.3969/j.issn.1673-8640.2017.01.006

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Objective To establish microscopy review rules of AVE automatic urine analysis. Methods Urine dry chemical and urinary sediment analysis was performed for 2 278 urine samples by AVE-752 urine dry chemical analyzer and AVE-766 urinary sediment analyzer. All samples were screened by microscopy with double blind method. AVE-752 and AVE-766 results,modified results and microscopy results were all recorded. Taking the average of microscopy results as standard,the review rules and review rates were established by false negative rates,false positive rates and error rates of 4 schemes（scheme 1：the results of AVE-752 and AVE-766 were analyzed directly;scheme 2：the modified results of AVE-752 and AVE-766 were analyzed;scheme 3：the positive results of scheme 1 were reviewed by microscopy and analyzed; scheme 4：the positive results of scheme 2 were reviewed by microscopy and analyzed）,which were based on leucocyte esterase （LEU）,occult blood （BLD） and protein （Pro）by AVE-752 and white blood cell（WBC）, red blood cell（RBC） and cast（Cast） by AVE-766 and/or modified. A total of 344 urine samples were determined to verify review rules. Results The false positive rates were 20.4%,13.5%,0.0% and 0.0%,the false negative rates were 8.9%,4.2%,8.9% and 4.2%,and the error rates were 16.8%,10.6%,2.8% and 1.3% . The review rates of scheme 3 and scheme 4 were 42.4% and 39.1%,108 samples with false positive results were modified （76 cases of WBC,23 cases of RBC and 9 cases of Cast）,and 33 samples with false negative results were modified（8 cases of WBC and 25 cases of RBC）. There still existed 30 samples with false negative results （12 cases of WBC and 18 cases of RBC）after modification. The false positive rate of 344 verified urine samples was 0.0%,the false negative rate was 4.5%,the error rate was 1.5%,and the review rate was 35.2%. Conclusions After the combined usage of AVE-752 and AVE-766,all samples should be screened by microscopy except LEU/WBC,BLD/RBC and Pro/Cast.

Eperythrozoon infection in patients with blood diseases

LU Lingyi, ZHAI Yuanqi, WANG Jianbiao

2017, 32(1):  26-29.  DOI: 10.3969/j.issn.1673-8640.2017.01.007

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Objective To investigate the infection rate of eperythrozoon in patients with blood diseases. Methods A total of 50 healthy subjects and 223 patients with blood diseases were enrolled. Eperythrozoon was determined by acridine staining. The patients with blood diseases were classified according to disease types and characteristics. The relationship between infection rate and blood diseases was analyzed. Results The infection rates in healthy subject group and blood disease group by acridine staining were 12.00% and 28.25%,respectively. The infection rate in blood disease group was higher than that in healthy subject group. No difference of infection rate was found between males and females（P>0.05）. People from 45 to 65 years old had the highest infection rate,and the infection rates showed no statistical significance for ages（P>0.05）. The infection rates between outpatients from hematology department and inpatients had no statistical significance（P>0.05）. The infection rate of patients with lymphoma was the highest in blood disease group. There was statistical significance for the infection rate between patients with lymphoma and patients with other blood diseases（P<0.05）. There was no statistical significance between patients with acute myeloid leukemia and patients with acute lymphocyte leukemia（P>0.05）. Conclusions Eperythrozoon mostly infects people from 45 to 65 years old,and there is no difference between males and females. Nosocomial infection is less likely to occur. Patients with blood diseases especially with lymphoma are more susceptible than healthy subjects to eperythrozoon. Different types of blood diseases have different infection rates. Thus,attention should be paid during treatment.

Correlation between EGFR gene mutations and clinical characteristics among lung cancer patients in Shanghai

JIANG Wenrong, CAI Yongjun, MIAO Yingxin, JIN Jingjing, XIAO Li, YANG Feng, YIN Yulei, ZHANG Yanmei, ZHAO Hu

2017, 32(1):  30-34.  DOI: 10.3969/j.issn.1673-8640.2017.01.008

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Objective To study the mutations of epidermal growth factor receptor （EGFR）gene and the correlation with the clinical characteristics of lung cancer patients in Shanghai. Methods A total of 207 paraffin-embedded tissue samples from lung cancer patients were collected. The mutations of EGFR gene were determined by real-time fluorescence quantitation polymerase chain reaction（PCR） after DNA extraction. Results The EGFR gene mutation rate was 47.34%,and there were correlations with sex and pathological types （P<0.01）,while there was no correlation with age （P>0.05）. The mutation types of EGFR gene were not correlated with sex and age （P>0.05）. EGFR gene mutation status of lung adenocarcinoma patients was not correlated with its pathological classification（P>0.05）. Conclusions EGFR gene mutations in Shanghai are correlated with the clinical characteristics of lung cancer patients. EGFR gene mutation occurs more frequently in females and in patients with lung adenocarcinoma.

Correlations of TNF-α gene polymorphism and its expression with primary hepatocellular carcinoma in Guangxi Zhuang population

LI Yanqiu, ZHU Bo, OU Chao, ZHAO Huiliu, SHU Hong, RONG Minhua

2017, 32(1):  35-40.  DOI: 10.3969/j.issn.1673-8640.2017.01.009

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Objective To investigate the correlations of tumor necrosis factor-alpha（TNF-α） -238/-308 single nucleotide polymorphism（SNP） and serum TNF-α levels with primary hepatocellular carcinoma （HCC） in Guangxi Zhuang population. Methods A total of 175 subjects were classified into primary HCC group（102 cases） and healthy control group（73 cases）. Serum TNF-α levels and the genotypes of TNF-α-238/-308 SNP were determined by double antibody sandwich enzyme-linked immunosorbent assay（ELISA） and polymerase chain reaction（PCR）-restriction fragment length polymorphism（RFLP）. Results Serum TNF-α levels in primary HCC group were higher than those in healthy control group（P<0.01）. For the genotype of TNF-α-308,serum TNF-α level of GA+AA genotype was higher than that of GG genotype（P<0.01）. The genotype and allele frequencies of TNF-α-308 in primary HCC group were higher than those in healthy control group（P<0.05）. The genotype and allele frequencies of TNF-α-238 in primary HCC group and healthy control group had no statistical significance（P>0.05）. The risk of primary HCC was higher in TNF-α-308 GA or AA genotype than that in TNF-α-308 GG genotype [odds ratio（OR）=3.556,95% confidence interval（CI）1.581-7.994]. The risk in TNF-α-308 A allele was 3.153 times higher than that in TNF-α-308 G allele （OR=3.153,95%CI 1.465-6.784）. Conclusions Serum TNF-α levels in primary HCC group are higher than those in healthy control group. TNF-α-308 SNP are correlated to serum TNF-α levels. TNF-α-308 SNP may be correlated to the genetic susceptibility of primary HCC in Guangxi Zhuang population.

Homocysteine,folic acid and vitamin B₁₂ for hypertension and its complications

BAI Ping, ZHU Jiang, YAO Yanlin, MA Jianhong, DUAN Falan, LI Jiufen

2017, 32(1):  41-44.  DOI: 10.3969/j.issn.1673-8640.2017.01.010

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Objective To investigate the clinical significance of homocysteine（Hcy）,folic acid and vitamin B₁₂ （Vit B₁₂） for hypertension and its complications. Methods A total of 4 358 subjects were enrolled and determined for Hcy,folic acid and Vit B₁₂,and other information of healthy examination was collected. According to hypertension and its complications,they were classified into simple hypertension group（653 cases）, hypertension with coronary heart disease（CHD） group（550 cases）, hypertension withs diabetes mellitus（DM） group（386 cases）,hypertension with other disease group（154 cases） and healthy control group（2 615 cases）. Results There were 1 743 cases with hypertension,and its determination rate was 39.99%. The levels of Hcy,Vit B₁₂ and folic acid in healthy control group were normal. The Hcy levels of simple hypertension group,hypertension with CHD group,hypertension with DM group and hypertension with other disease group increased linearly,and the levels of Vit B₁₂ and folic acid decreased linearly. Compared to healthy control group,age,body mass index（BMI）,sex proportion,the rates of tobacco and alcohol abuses,Hcy level and hyperhomocysteinemia rate of the other 4 groups were higher（P<0.05）,and the levels of folic acid and Vit B₁₂ were lower（P<0.05）. Compared to simple hypertension group,the other hypertension groups had higher level of Hcy and hyperhomocysteinemia rate（P<0.05） and lower levels of folic acid and Vit B₁₂（P<0.05）. Compared to hypertension with CHD group,hypertension with DM group and hypertension with other disease group had higher level of Hcy（P<0.05） and lower levels of folic acid and Vit B₁₂（P<0.05）. There was no statistical significance of other comparisons among groups（P>0.05）. Conclusions High level of Hcy,low level of Vit B₁₂ and low level of folic acid are risk factors for hypertension and its complications. Hypertension is associated with age,tobacco and alcohol abuses and BMI.

Serum carbohydrate deficient transferrin level of males with different nationalities in Xinjiang

AERZIGULI· Mutalifu, LI Xiaoqin, WANG Changmin

2017, 32(1):  45-47.  DOI: 10.3969/j.issn.1673-8640.2017.01.011

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Objective To study serum carbohydrate deficient transferrin（CDT）level of males with different nationalities in Xinjiang,and to assess the role of CDT in identifying alcohol abuse among males. Methods A questionnaire survey was conducted,and 131 healthy subjects （healthy control group,including 53 cases of Han,46 cases of Uygur and 32 cases of Kazak） and 96 subjects with alcohol abuse （alcohol abuse group,including 45 cases of Han,31 cases of Uygur and 20 cases of Kazak）were enrolled. Serum CDT level was determined by capillary electrophoresis. The reference interval and cut-off value were determined by P-P normality test and receiver operating characteristic（ROC） curve,respectively. Results The CDT level of alcohol abuse group was 3.530%±0.833%,which was higher than that of healthy control group （0.716%±0.341%,P<0.001）. There was no statistical significance for different nationalities in 2 groups（P>0.05）. The reference interval for serum CDT level in healthy control group was <1.28%. According to ROC curve,the cut-off value for alcohol abuse group was 1.25%,the sensitivity was 81.3%,and the specificity was 92.3%. The area under ROC curve was 0.948. Conclusions The reference interval of serum CDT level in healthy subjects with different nationalities in Xinjiang could use the same level. The determination of CDT has good sensitivity and specificity for identifying alcohol abuse,which meets clinical needs.

Alkali treatment method for improving the specificity of serum 1,3-beta-D-glucan determination

XUE Zhixin, LIU Juanjuan, WANG Baoxue, LI Yanan, SU Yan, ZHOU Zeqi

2017, 32(1):  48-51.  DOI: 10.3969/j.issn.1673-8640.2017.01.012

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Objective To study alkali treatment method for improving the specificity of serum 1,3-beta-D-glucan determination. Methods A total of 73 patients suspected with invasive fungal disease were enrolled from April 2013 to December 2014. Serum 1,3-beta-D-glucan level was determined by alkali treatment method and heating dilution method,respectively. The results of fungal culture were as evaluation standard. The results were analyzed by SPSS 19.0 software. Results The coincidence rates of alkali treatment method and heating dilution method with fungal culture were 87.50% and 63.64%,respectively. There was statistical significance between alkali treatment method and heating dilution method（P<0.05）. The consistency of the 2 treatment methods was not good（Kappa=0.718）. Conclusions Alkali treatment method could make 1,3-beta-D-glucan existing with a low structure form and also eliminating the effect of high structure,which improves the specificity of serum 1,3-beta-D-glucan determination. So, alkali treatment method is better than heating dilution method.

Influence of LOX gene drop expression in human vaginal wall fibroblasts on the mRNA expressions of ELN and MMP-2

ZHOU Qian, WU Huiheng, CHEN Xinliang

2017, 32(1):  52-56.  DOI: 10.3969/j.issn.1673-8640.2017.01.013

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Objective To study the influence of lysyl oxidase（LOX） gene drop expression in human vaginal wall fibroblasts on the mRNA expressions of elastin （ELN） and matrix metalloproteinase-2（MMP-2）. Methods The primary-cultured human vaginal wall fibroblasts were classified into 3 groups,LOX gene drop expression group,negative control group and blank control group. LOX-shRNA lentiviruses were transfected into the fibroblasts of LOX gene drop expression group. Negative control lentiviruses were transfected into the fibroblasts of negative control group. The transfection efficiency was calculated. Flow cytometry sorted fibroblasts with green fluorescence,and some cells were collected and determined. Real-time fluorescence quantitation polymerase chain reaction（PCR） was used to determine the mRNA expressions of LOX,ELN andMMP-2. Results A total of 8 samples were expressed in the fibroblasts in LOX gene drop expression group,including 4 cases of premenopausal women and 4 cases of postmenopausal women. The fibroblasts in LOX gene drop expression group became smaller,the growth was slow,and the adherence decreased. The expression of LOX mRNA in LOX gene drop expression group was lower than those in negative and blank control groups （P<0.05）. The expression of ELN mRNA in LOX gene drop expression group was lower than those in negative and blank control groups,while the expression of MMP-2 mRNA was higher. There was no statistical significance for LOX mRNA,ELN mRNA and MMP-2 mRNA expressions between negative and blank control groups（P>0.05）. Conclusions The expression of ELN mRNA decreases,however,that of MMP-2 mRNA increased. LOXmay participate in the occurrence of pelvic organ prolapse（POP） by regulating the expressions of ELN mRNA and MMP-2 mRNA.

MALDI-TOF MS in the determination of drug resistance

YANG Feng, CHEN Fei, ZHAO Hu

2017, 32(1):  57-63.  DOI: 10.3969/j.issn.1673-8640.2017.01.014

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The application of matrix-assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF MS） has brought a revolutionary change for microbial identification program in clinical microbiological laboratories,for its fast,high-throughput,accuracy and cost-effectiveness. Furthermore,MALDI-TOF MS provides a new way for drug resistance determination. It is time-consuming for commonly-used methods of routine drug resistance determination,such as Kirby-Bauer,dilution and E-test. These methods can not meet the clinical demand for quick selection of sensitive drugs. So,the application of MALDI-TOF MS for the determination of drug resistance has research value and prospect. Some experiments and clinical application of MALDI-TOF MS for determining drug resistance have been reported. This review introduces the types of pathogens,determination methods and the efficiency of MALDI-TOF MS for determining drug resistance in recent years.

Delta checks based on individual patients' data

FEI Yang, WANG Wei, WANG Zhiguo

2017, 32(1):  64-68.  DOI: 10.3969/j.issn.1673-8640.2017.01.015

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Risks are inherent in each step of whole testing process. Delta checks based on individual patients' data are methods for mitigating these risks. Delta checks can be used as a complementary tool for traditional laboratory quality control to determine issues that quality control materials may not address. It can be applied into different phases in whole testing process,including determining misidentified and mishandled specimens,true patients' changes,instrumentation and personal errors,as well as the evaluation of point-of-care testing（POCT）. The review introduces the application of delta checks in POCT and pre-examination,intra-examination and post-examination phases,including selecting candidate analytes,establishing monitor parameters,setting limits and the evaluation of usefulness,hoping to provide a reference for clinical laboratories.