Table of Content

30 November 2018, Volume 33 Issue 11

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Orginal Article

Correlations between IgG,IgM and IgA anti-ds-DNA antibodies and systemic lupus erythematosus

SONG Rui, YE Ping, CHEN Xiaoxiang

2018, 33(11):  969-974.  DOI: 10.3969/j.issn.1673-8640.2018.011.001

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Objective To investigate the correlations of 3 subtypes（IgG,IgM and IgA） of anti-double-stranded DNA（ds-DNA） antibodies with the disease activity of systemic lupus erythematosus（SLE） and SLE clinical phenotypes. Methods A total of 202 SLE patients were enrolled. The 3 subtypes（IgG,IgM and IgA） of anti-ds-DNA antibodies were determined by enzyme-linked immunosorbent assay（ELISA）,and the correlations of IgG,IgM and IgA anti-ds-DNA antibodies with 15 clinical phenotypes and systemic lupus erythematosus disease activity index（SLEDAI） were evaluated. Results The levels of IgG,IgM and IgA anti-ds-DNA antibodies in SLE group were higher than those in healthy control group（P<0.01）. The levels of IgM, IgA and IgG anti-ds-DNA antibodies in SLE group were from low to high in turn（P<0.01）,which were higher in SLE active group than those in SLE remission group（P<0.01）. However, the positive rates of IgG,IgM and IgA anti-ds-DNA antibodies between the 2 groups had no statistical significance（P>0.05）. Spearman correlation analysis showed that there were positive correlations of IgG, IgM and IgA anti-ds-DNA antibodies with SLEDAI in SLE group（r_s=0.446,0.249 and 0.507,P<0.01）. Clinical phenotypes,including anti-nucleosome antibody positive,anti-nuclear antibody positive,proteinuria,complement decreasing,peripheral arthritis,malar rash and alopecia,were correlated with IgG anti-ds-DNA antibody（P<0.05,P<0.01）. Anti-nucleosome antibody positive,complement decreasing,leukopenia,pleuritis and fever were correlated with IgM anti-ds-DNA antibody（P<0.05,P<0.01）. Anti-nucleosome antibody positive,anti-nuclear antibody positive,complement decreasing, peripheral arthritis and leukopenia were correlated with IgA anti-ds-DNA antibody（P<0.05,P<0.01）. Conclusions The levels of IgG and IgA anti-ds-DNA antibodies are correlated with the disease activity of SLE,while IgM has less effect in SLE. Clinical phenotypes such as anti-nucleosome antibody positive and complement decreasing show correlations with the 3 subtypes.

Pathogenic distribution and drug susceptibility characteristics of lower respiratory tract infection of children in central and southern Sichuan

HU Rong, NIE Bin, XIANG Xiaojie, DING Guimei, LIU Nengying, YANG Xueqiang

2018, 33(11):  975-978.  DOI: 10.3969/j.issn.1673-8640.2018.011.002

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Objective To review the common pathogens and drug susceptibility of lower respiratory tract infection（LRI） of children,and to provide a reference for the rational use of drugs in clinic. Methods A total of 6 205 patients with LRI were enrolled,the isolated strains through sputum culturing were identified and determined by VITEK 2 Compact automatic microbiology analyzer,and the drug susceptibility was performed. Results From 2014 to 2016,14 species of pathogens （2 147 isolates）were isolated from sputum samples,and the positive rate was 34.6%. The main species were Streptococcus pneumoniae （39.17%,840 isolates）,Haemophilus influenzae （23.43%,503 isolates）,Moraxella catarrhalis（20.63%,443 isolates） and Staphylococcus aureus （7.59%,163 isolates）. Streptococcus pneumoniae had a decreased trend,while Moraxella catarrhalis had an increased trend. The drug resistance rates of penicillin,sulfamethoxazole and other antibiotics were increasing. Conclusions The pathogenic distribution of children with LRI in central and southern Sichuan has its own characteristics. Clinicians should adjust drugs according to the drug resistance changes of pathogens and reduce the risk of drug resistance caused by the improper use of drugs.

CYP2C19 gene polymorphism in the drug therapy of coronary heart disease

ZHANG Shaohong, SHEN Xuebin

2018, 33(11):  979-982.  DOI: 10.3969/j.issn.1673-8640.2018.011.003

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Objective To study the distribution characteristic of CYP2C19 gene polymorphism in coronary heart disease （CHD）patients in northern Fujian,to evaluate the clinical prognosis of different anti-platelet （PLT） drug therapies after percutaneous coronary intervention（PCI）,and to provide a reference for anti-PLT aggregation treatment. Methods A total of 520 patients diagnosed as CHD by coronary angiography undergoing PCI were enrolled and followed up. CYP2C19 genotype was determined. All patients should be in drug-eluting stents（DES）. According to genotypes combined with coronary artery disease severity,the patients were classified into 3 groups（fast,intermediate and slow metabolic types）. The distribution of genotypes was assessed. After PCI for 1,6 and 12 months,the occurrence of major adverse cardiovascular events （MACE） was recorded. Results There were 224 （43.07%） cases of CYP2C19 homozygous fast metabolic type. There were 227 （43.65%） cases of intermediate metabolic type,including 186 （82%） cases of 681 base function locus mutation. There were 69 （13.26%） cases of slow metabolic type. After 1-year follow-up,there was no statistical significance for MACE occurrence among the 3 groups （P>0.05）. Conclusions CYP2C19 loss-of-function （LOF） increases in patients with CHD in northern Fujian. Through genotype determination,selecting different anti-PLT aggregation drugs or adjusting anti-PLT aggregation drug doses,the occurrence of MACE can decrease,which can improve long-term prognosis in CHD patients.

Evaluation on the real-time correction of whole blood CRP determination using HCT value

MA Jirong, ZHOU Jingyi, GU Yi, SHEN Wenyan, SHEN Wei

2018, 33(11):  983-986.  DOI: 10.3969/j.issn.1673-8640.2018.011.004

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Objective To evaluate the real-time correction of whole blood C-reactive protein （CRP） determination using hematocrit （HCT） value in order to ensure the accuracy of determination results. Methods A total of 217 blood samples were collected. HCT<35% was found in 37 cases,HCT 35%-45% in 128 cases,and HCT>45% in 52 cases. The comparability of CRP levels before and after correcting in XN 2800 was evaluated. Results The mean deviation between uncorrected level and target CRP level by BN Ⅱ specific protein analyzer was 11.77 mg/L in 37 cases with HCT<35%,and the deviation was negatively correlated with the decrease of HCT,while the mean deviation between corrected and target CRP levels was 2.75 mg/L. In 128 cases with HCT 35%-45%,the mean deviation between uncorrected and target CRP levels was 2.79 mg/L,while the mean deviation between corrected and target CRP levels was 2.77 mg/L. In 52 cases with HCT>45%,the mean deviation between uncorrected and target CRP levels was -11.84 mg/L,and the deviation was positively correlated with the increase of HCT,while the mean deviation between corrected and target CRP levels was -1.35 mg/L. Conclusions As to blood samples with HCT<35% and HCT>45%,real-time correction of whole CRP value determination using HCT value can make the determination results more accurate.

Relationship between apolipoprotein E gene polymorphism and blood lipid metabolism in hyperlipidemia patients

LIU Huifang

2018, 33(11):  987-990.  DOI: 10.3969/j.issn.1673-8640.2018.011.005

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Objective To study the relationship between apolipoprotein E （ApoE）gene polymorphism and blood lipid metabolism disorder,and to guide rational drug use. Methods A total of 142 hyperlipidemia patients were enrolled from January 2016 to December 2016. Patients with one of total cholesterol （TC）,triglyceride （TG） and low-density lipoprotein cholesterol （LDL-C）increasing were enrolled. The genotypes of ApoE gene were detected by gene chip technology. The relationship between different genotypes with blood lipid metabolism was evaluated. Results Among the 142 hyperlipidemia patients,there were 51 cases of high TC,70 cases of high TG and 49 cases of high LDL-C. ApoEgene showed polymorphic distribution. The distribution frequency of E3 allele was the highest,accounting for 65.42%,followed by E4 （17.53%） and E2 （17.05%）. The levels of TC and LDL-C in E4 group were higher, and the TG level in E2 group was higher. There was no statistical significance for TG and LDL-C levels among the 3 genotypes （P>0.05）. E4 genotype had statistical significance for TC levels compared with E2 and E3 genotypes （P<0.05）. Conclusions ApoEgene presents polymorphic distribution. Hyperlipidemia is an independent factor for atherosclerosis and cardiovascular diseases. Blood lipid levels are influenced from different genotypes. Targeting different ApoE gene genotypes, individualized treatment programs may be developed for patients, in order to select appropriate lipid-lowering drugs.

Chitinase-3-like protein 1 combined with estimated glomerular filtration rate in evaluating the curative effect of interventional therapy for atherosclerotic renal artery stenosis

QIN Chuntang

2018, 33(11):  991-996.  DOI: 10.3969/j.issn.1673-8640.2018.011.006

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Objective To evaluate the roles of chitinase-3-like protein 1（CHI3L1） and estimated glomerular filtration rate（eGFR） in evaluating the curative effect of interventional therapy for atherosclerotic renal artery stenosis（ARAS）. Methods The clinical data of 78 patients with ARAS treated by percutaneous renal artery stenting （PRAS） were collected. According to the results of CHI3L1 and eGFR at 1 month after treatment,the patients were classified into group A （CHI3L1 and eGFR were abnormal,20 cases）,group B（CHI3L1 or eGFR was normal,28 cases） and group C（CHI3L1 and eGFR were normal,30 cases）. The baseline data （sex,age,the duration of hypertension,the type of antihypertensive drugs,the location of renal artery stenosis,smoking and chronic diseases）,blood pressure in follow-up period,1 month before treatment and 1 month after treatment,urinary albumin/creatinine ratio（ACR）,serum creatinine（SCr） level and the degree of renal artery stenosis were compared among the 3 groups. The incidence rates of adverse events and survival rates among the 3 groups in follow-up period were recorded. Results There was no statistical significance among the 3 groups for the baseline data and follow-up period（P>0.05）,and the CHI3L1 levels of group A,B and C decreased in turn（P<0.05）. There was no statistical significance in blood pressure,urinary ACR,SCr level and the degree of renal artery stenosis among the 3 groups before treatment （P>0.05）. After treatment,diastolic blood pressure,systolic blood pressure,urinary ACR,SCr level,CHI3L1 level and the degree of renal artery stenosis decreased gradually （P<0.05）. The incidence rates of adverse events in group A,B and C were 23.33%,17.86% and 6.67%,respectively,and the 1-year survival rates after treatment were 75.00%,92.86% and 100.00%,respectively（P<0.05）. The sensitivity and specificity of CHI3L1 combined determination with eGFR in the evaluation of interventional therapy were 95.65% and 55.56%,which were higher than those of eGFR determination （94.20% and 11.11%）. Conclusions CHI3L1 combined determination with eGFR is of value in evaluating the curative effect of interventional therapy and the prognosis of ARAS.

Role of Semaphorin 3A in the early diagnosis of contrast-induced acute kidney injury

ZHANG Yixin, WEI Dianjun, NING Li

2018, 33(11):  997-1003.  DOI: 10.3969/j.issn.1673-8640.2018.011.007

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Objective To investigate the role of Semaphorin（Sema） 3A in the early diagnosis of contrast-induced acute kidney injury（CIAKI）. Methods A total of 350 patients undergoing coronary angiography and stent implantation were enrolled and classified into 2 groups,CIAKI group（46 cases） and non-CIAKI group（304 cases）. Serum and urinary Sema 3A and serum beta₂-microglobulin（β₂-MG）,cystatin C（Cys C） and serum creatinine（SCr）were determined before surgery and at 6,12,24,48 and 72 h after surgery. Receiver operating characteristic（ROC） curve was performed to evaluate the roles of these parameters for the diagnosis of CIAKI. Spearman correlation analysis was used to identify the relationship between the levels of urinary Sema 3A in different times and clinical manifestation [the percentage changes of SCr,hospitalization time after surgery and the duration of acute kidney injury（AKI）]. Logistic regression analysis was utilized to assess the predictive factors of AKI. Results Compared with preoperative baseline values,SCr,β₂-MG and Cys C in CIAKI group increased to different degrees at 24,48 and 72 h after surgery（P<0.05）,and there was statistical significance compared with those in non-CIAKI group（P<0.05）. Cys C had the best diagnostic performance at 48 h after surgery. The levels of serum Sema 3A in CIAKI group increased at 12,24 and 48 h after surgery,and urinary Sema 3A increased at 6 h after surgery. Compared with β₂-MG,Cys C and SCr,urinary Sema 3A in CIAKI group was increased earlier. Correlation analysis showed that the level of urinary Sema 3A before surgery was negatively correlated with age,the change of SCr after surgery and the duration of AKI（P<0.05）. The high levels of urinary Sema 3A at 6 and 12 h after surgery were correlated with the increasing of SCr,hospitalization time and the duration of AKI（P<0.01）. Logistic regression analysis showed that sex and the level of urinary Sema 3A at 6 h after surgery were independent predictive factors of CIAKI. ROC curve analysis showed that urinary Sema 3A at 6 h after surgery had optimal diagnostic performance in the diagnosis of CIAKI,and the optimal cut-off value was 245 pg/μmol Cr. The sensitivity,specificity,positive predictive value and negative predictive value was 84%,92%,91% and 87%,respectively. Conclusions Sema 3A is closely associated with the occurrence of early kidney injury,which is an early biomarker of CIAKI.

Performance verification of CS-5100 automatic blood coagulation analyzer for the determinations of antithrombin Ⅲ,protein C and protein S activities

LI Wen, HE Aili, ZHANG Wen, ZHOU Ting

2018, 33(11):  1009-1012.  DOI: 10.3969/j.issn.1673-8640.2018.011.009

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Objective To evaluate the performance of CS-5100 automatic blood coagulation analyzer for the determinations of antithrombin Ⅲ（AT Ⅲ）,protein C（Pc） and protein S（Ps） activities. Methods According to the Clinical and Laboratory Standards Institute （CLSI） guidelines,the imprecision [coefficient of variation （CV）],accuracy,linearity,carry-over rate and reference interval of CS-5100 for the determinations of AT Ⅲ,Pc and Ps activities were validated. Results The within-run CV of CS-5100 for the determinations of AT Ⅲ,Pc and Ps activities were <1.9%,and the inter-day CV were <4.1%,meeting manufacturers' requirements. The bias of accuracy verification results was within biology CV requirements. The linear regression equation between AT Ⅲ linear validation theoretical value and measured values met the slope of 1±0.05,the determination coefficient （r²）≥0.95 and carry-over CV<10%. Reference interval verification results showed that the determination index R was ≥0.9. Conclusions The precision,accuracy,linearity,carry-over rate and reference interval of CS-5100 meet the requirements of quality control,which can ensure determination quality.

Determination of human serum ouabain by UPLC-MS/MS and methodology comparison

ZHAO Yinxia, OU Meixian, QU Yi, LU Youli, ZHANG Meiwei, LI Shuijun

2018, 33(11):  1013-1016.  DOI: 10.3969/j.issn.1673-8640.2018.011.010

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Objective To establish an ultra high performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS） for the determination of human serum ouabain. Methods High-specific UPLC-MS/MS was used. A deuterium-labeled ouabain-d3 was used as internal standard. Serum samples were prepared with solid phase extraction（SPE）,and a reversed phase column was used for chromatography separation. The mass spectrometer was operated in negative-ion mode with electrospray ionization（ESI） source. Methodology validation included the evaluation of matrix effect,recovery,accuracy,within-run precision,between-run precision and stability. The established UPLC-MS/MS was used to determine serum ouabain in 40 hypertension patients and 20 healthy subjects （healthy control group）. The results of UPLC-MS/MS were compared with those of enzyme-linked immunosorbent assay （ELISA）. Results The UPLC-MS/MS had a linear range from 0.02 to 5.00 ng/mL. The lower limit of quantification （LLOQ） was 0.02 ng/mL. The matrix effect of pretreatment with ABN solid phase extraction column was the smallest. The recovery rate was high （85%）. The accuracies of LLOQ,low-level （0.06 ng/mL）,middle-level （0.6 ng/mL） and high-level （4 ng/mL） quality control materials were 108.0%,89.2%,101.0% and 103.0%,respectively. The within-run coefficients of variation （CV） were 2.87%,1.95% and 0.56%,and the between-run CV were 5.98%,1.90% and 0.75%. The samples placed at room temperature for 16 h were stable,and the processed samples placed in autosampler at room temperature for 48 h were stable. Both biases were <15%. The results showed that both healthy control and hypertension groups had undetectable serum ouabain level by the established UPLC-MS/MS. The levels were 0.096 ng/mL in hypertension group and 0.062 ng/mL in healthy control group when they were determined by ELISA. Ouabain standard materials at 5 different levels （0.02,0.05,0.10,0.20 and 0.50 ng/mL） were determined by UPLC-MS/MS. It showed a positive correlation between the results and the corresponding levels with good linearity（r²>0.99）and high accuracy. The results of 5 samples determined by ELISA were very close（0.024 9-0.029 6 ng/mL）. Conclusions The level of serum ouabain is undetectable both in healthy control and hypertension groups by the established UPLC-MS/MS. There is difference for the results of serum ouabain determination between UPLC-MS/MS and ELISA.

Establishment and performance evaluation of candidate reference measurement procedure for serum total homocysteine by liquid chromatography-tandem mass spectrometry

SHEN Min, YANG Xiaodong, WANG Lin, ZOU Jihua, ZHANG Man, ZOU Bingde

2018, 33(11):  1018-1025.  DOI: 10.3969/j.issn.1673-8640.2018.011.011

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Objective To establish a candidate reference measurement procedure for serum total homocysteine （Hcy） by liquid chromatography-tandem mass spectrometry （LC-MS/MS）,and to evaluate its analytical performance. Methods Samples were prepared by a simple protein precipitation method,and LC-MS/MS was used to determine serum total Hcy quantitatively. According to the Clinical and Laboratory Standards Institute（CLSI） C62-A and C50-A documents,the performance,including linearity,limit of detection,limit of quantitation,relative matrix effect,precision and trueness,was evaluated. Results The linear range of total Hcy by LC-MS/MS was 0.5-200.0 μmol/L. The limit of quantitation and limit of detection were 0.31 nmol/g and 0.06 nmol/g,respectively. The relative matrix effects were 1.94%,1.91% and 1.78% for sera and solution mixtures in 3 different ratios（1∶1,80∶20 and 20∶80）. The within-run and between-run coefficients of variation（CV）were <2% and <1%. The average recovery rates of 3 levels （30.58,49.21 and 65.42 nmol/g） were 99.8%,100.2%,100.8%, respectively. The bias of NIST standard reference material （SRM） 1950 was <1%. After the samples were processed,they were placed at room temperature [（23±2）℃] and autosampler（10℃） for 24 h, and the results were stable. Conclusions The candidate reference measurement procedure for serum total Hcy by LC-MS/MS has been established,and it has good accuracy and precision,which can be used for measurement traceability.

Analysis on the results of gamma-glutamyltransferase trueness verification survey in Shanghai

YU Xiaoxuan, OU Yuanzhu, TANG Liping, WANG Meijuan, LIU Wenbin, GE Danhong, WANG Ming

2018, 33(11):  1032-1035.  DOI: 10.3969/j.issn.1673-8640.2018.011.014

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Objective To trace the terminal results of gamma-glutamyltransferase （GGT） determination in clinical laboratories to enzymological international system of units through determining GGT trueness control samples assigned the International Federation of Clinical Chemistry and Laboratory Medicine （IFCC） primary reference method,and to achieve the comparability of GGT determination results among clinical laboratories. Methods Four trueness control samples after centrifugation and filtration were transported to clinical laboratories under cold chain,and the samples were determined for 3 d continuously. The trueness control samples were assigned using IFCC primary reference method by Shanghai Center for Clinical Laboratory Enzymological Reference Laboratory,and the target values were obtained. Each sample was not grouped,and the x,s,coefficients of variation （CV） and biases against target values of the 2 samples were evaluated. After each sample was grouped according to instrument brands,the x,s,CV and biases against target values were evaluated as well. Results The results of 82 clinical laboratories showed that the overall CV of the 2 samples were <10%,and the biases against target values were <7.3%. The 82 clinical laboratories were classified into 8 groups according to instrument brands,the within-group CV and biases against target values in Roche,Beckman and Mindray groups were <5.6%. The within-group CV of Beckman,Hitachi and Toshiba groups were <6.4%. The biases of Hitachi and Toshiba groups were >7.2% for A-level sample. The biases for B-level sample was <5.7%. The bias of Beckman group was >30%. The CV of Siemens and Abbott groups were about 10%,and the biases were about 6%. With the target value ±5.5%,the total qualified coincidence rate did not meet the standard of 61%. Conclusions The bias between the results of GGT determination by clinical laboratories tracing back to IFCC reference method and the results assigned using IFCC reference method is small,but the bias of the results of GGT determination by clinical laboratories in Beckman group with IFCC reference method is big.

Research progress in the determination methods of tumor markers

ZHANG Yingcong, ZHANG Ze, YU Hongwei, SANG Zhuoqi, CHANG Dong

2018, 33(11):  1036-1042.  DOI: 10.3969/j.issn.1673-8640.2018.011.015

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Tumor is a serious hidden threat to human health,and the morbidity and mortality rates of the diseases caused by tumors are increasing year by year. Tumor markers are produced by tumor tissues or their host reacts,which can reflect and monitor the occurrence,development,prognosis and recurrence of tumors. Early determination of tumors is important,and monitoring the change of tumor markers is also an important method to evaluate the therapeutic effect of tumors. The common determination methods of tumor markers include immunology,biosensor,proteomics,molecular biology,liquid biopsy and so on. Different determination methods of tumor markers have different characteristics. This review focuses on the research progress of several common determination methods of tumor markers.

Research progress in laboratory diagnosis of IgG4-related diseases

SHU Jie, SHENG Huiming

2018, 33(11):  1043-1048.  DOI: 10.3969/j.issn.1673-8640.2018.011.016

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IgG4-related disease （IgG4-RD） is a newly recognized autoimmune disease characterized by fibrotic inflammation. The pathological features are IgG4⁺ plasma cell infiltration,tabular fibrosis and occlusive phlebitis,and IgG4-RD was accompanied with elevated serum IgG4 levels. Most human organs can be affected,which can result in anthorisma due to chronic inflammation and fibrosis. Descendiblity,immue,microbial infection and molecular simulation are involved in its pathogenesis. At present,pathological diagnosis is still the gold standard for the diagnosis of IgG4-RD,but laboratory serological determination and immune cell analysis have the advantages of quantitation,convenience and accuracy,and they are irreplaceable. This review focuses on the naming,pathogenesis and laboratory diagnosis of IgG4-RD.

Research progress of exosomal miRNA in pancreatic cancer

CHEN Jia, WANG Jian, JIN Wei, CHEN Xudong, HU Xiaobo

2018, 33(11):  1049-1052.  DOI: 10.3969/j.issn.1673-8640.2018.011.017

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Exosomes are sorts of small extracellular vesicles ranging approximately from 30 nm to 100 nm in diameter,which can serve as carriers transporting microRNA（miRNA）,protein and others to target cells by means of cell membrane fusion and cell swallowing,further mediating cell communication and regulating cell physiological function. In recent years,the related researches have found that exosomal miRNA plays a role in the occurrence,development and clinical diagnosis of pancreatic cancer（PC）,which is expected to provide new ideas for the diagnosis and treatment of PC. In this review,the role of exosomal miRNA in PC will be introduced based on domestic and overseas research status.