Table of Content

30 December 2021, Volume 36 Issue 12

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Expert consensus on establishment of standards for the performance evaluation of systems for whole blood C-reactive protein determination of Chinese children

Clinical Diagnosis and Laboratory Medicine Branch of China Maternal and Child Health Association

2021, 36(12):  1201-1205.  DOI: 10.3969/j.issn.1673-8640.2021.012.001

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C-reactive protein（CRP）,which can be determined by serum as well as whole blood,has been traditionally utilized as a marker of infection. Many brands of whole blood CRP determination equipments have been widely used in medical institutions,especially in women's and children's hospitals. However,the standard for the performance evaluation of whole blood CRP determination system is not established. According to the data of 26 women's and children's hospitals,this consensus aims to provide the performance status of whole blood CRP determination system,and further put forward the performance evaluation requirements of whole blood CRP determination system in combination with clinical needs,biological variation and industry standards,so as to ensure the determination quality.

Analysis of cut-off values of MCV and MCH for screening thalassemia in adults in Dehong,Yunnan

SANG Guoxin, YI Wei, YANG Biqing, GUO Weixia, HUANG Kai, LIN Keqin, CHU Jiayou, YANG Zhaoqing

2021, 36(12):  1206-1209.  DOI: 10.3969/j.issn.1673-8640.2021.012.002

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Objective To investigate the optimal cut-off values of mean corpuscular volume（MCV） and mean corpuscular hemoglobin（MCH） for screening thalassemia in adults in Dehong,Yunnan. Methods Totally,9 562 adults undergoing outpatient examination,physical examination,premarital examination and genetic counseling in the People's Hospital of Dehong Dai and Jingpo Autonomous Prefecture were enrolled,and they were classified into thalassemia group（2 662 cases） and healthy control group（6 900 cases） through the determination of common gene mutations of thalassemia. Receiver operating characteristic（ROC） curve was drawn based on the MCV and MCH results obtained from routine blood test in order to determine the optimal screening cut-off values and efficiencies. Results The optimal cut-off value of MCV was 83.95 fL,the screening sensitivity was 82.7%,and the specificity was 89.7%. The optimal cut-off value of MCH was 28.15 pg,the screening sensitivity was 88.7%,and the specificity was 89.2%. Compared with the cut-off values widely used in China（MCV<80 fL and MCH<27 pg）,the screening cut-off values in this study may reduce the missed diagnosis rate,and the missed diagnosis rate of screening with MCV and MCH combinedly was the lowest. Conclusions Epidemic areas with unique mutation profiles may establish their screening cut-off values,and the combined screening of MCV or MCH may reduce the missed diagnosis rate and improve screening efficiency.

Role of time to positive of blood culturing in preliminary microbial identification

ZHAO Yanan, CAO Qixin, ZHAO Jianping

2021, 36(12):  1210-1214.  DOI: 10.3969/j.issn.1673-8640.2021.012.003

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Objective To study the role of time to positive（TTP） of blood culturing in preliminary identification of microbial species and distinguishing between pathogenic bacteria and contaminated bacteria,and to provide a reference for early clinical warning and rational drug use. Methods Totally,534 positive and non-repetitive isolates from bloodstream infection（BSI） patients in Inner Mongolia Autonomous Region People's Hospital from 2017 to 2020 were analyzed. A comprehensive analysis was conducted combined with clinical data. Results Among the 534 isolates from blood culturing,323 isolates（60.5%） were determined with TTP≤1 d,454 isolates（85.0%） were determined with TTP≤2 d,and 513 isolates（96.0%） were determined with TTP≤3 d. The median alarm times of positive blood culturing were different,from the fastest to the slowest which were Enterobacteriaceae（0.57 d）,Streptococcus（0.68 d）,non-fermentative bacterium（0.71 d）,Enterococcus（0.72 d）,Staphylococcus（1.36 d）,yeast（1.98 d） and Brucella melitensis（2.75 d）. There was no statistical significance between Enterobacteriaceae TTP and non-fermentative bacterium TTP,Streptococcus TTP（P>0.05）,but there was statistical significance between Enterobacteriaceae TTP and Enterococcus,Staphylococcus,yeast and Brucella melitensis（P<0.05）. The determination rate of contaminated bacteria in 534 isolates of blood culturing was 18.5%（99 isolates）,and the determination rates of contaminated bacteria were 15.2% in 1 d,46.5% in 1-2 d,26.2% in 2-3 d and 12.1% in >3 d. The TTP≥1 d of suspicious contaminated bacteria accounted for 84.8%. Coagulase-negative Staphylococcus（59.6%） and Gram-positive bacilli（9.1%） were the predominant bacteria in contaminated bacteria. The TTP between pathogenic bacteria and contaminated bacteria had statistical significance（P<0.05）. Conclusions Pathogenic bacteria and their complex distribution in BSI patients can be preliminarily predicted according to TTP,pathogenic bacteria and contaminated bacteria can be distinguished,and drugs can be used rationally.

Role of procalcitonin in the diagnosis of different sepsis

PAN Xilong, XU Zhiyuan, XIE Feng, LI Dan

2021, 36(12):  1215-1218.  DOI: 10.3969/j.issn.1673-8640.2021.012.004

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Objective To investigate the specific changes in the diagnostic efficacy of procalcitonin（PCT） when the diagnostic criteria for sepsis changed from Sepsis-1 to Sepsis-3,and to provide a reference for the clinical application of PCT in the diagnosis of sepsis. Methods A total of 189 patients with infection or（and） the manifestations of systemic inflammatory response syndrome（SIRS）（non-sepsis,sepsis and septic shock groups） and 30 healthy subjects（healthy control group）were enrolled. PCT was determined in all the subjects. Receiver operating characteristic（ROC） curves were used to evaluate the roles of Sepsis-1 and Sepsis-3 for the diagnosis of PCT. Results There was an increasing trend in PCT levels from healthy control,non-sepsis,sepsis to septic shock groups in each diagnostic component（P<0.05）. Based on Sepsis-1 and Sepsis-3 diagnostic criteria,the areas under curves were 0.85 and 0.73,the sensitivities were 76.85% and 57.01%,the specificities were 78.38% and 76.79%,respectively. The areas under curves,sensitivities and specificities of PCT in the diagnosis of septic shock were 0.75 and 0.76,73.91% and 64.86%,80.65% and 81.43%,respectively. Conclusions When the diagnostic criteria for sepsis change from Sepsis-1 to Sepsis-3,the diagnostic efficacy of PCT decreases in the diagnosis of sepsis,and the diagnostic efficacy of PCT does not change in the diagnosis of septic shock.

Application of fluorescent staining in fungus detection

ZHAO Lin, QIAO Yun

2021, 36(12):  1219-1221.  DOI: 10.3969/j.issn.1673-8640.2021.012.005

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Objective To evaluate the application of fluorescent staining in fungus detection. Methods A total of 212 clinical samples were collected. Fluorescent staining,direct smear,Gram staining,ink staining,fungal culturing and KOH wet film were used. Results In 175 deep samples,including sputum,pharyngeal swab,leucorrhea,urine and feces,the positive rates of fungi by fluorescent staining,direct smear and Gram staining were 87.43%,78.86% and 74.29%,respectively. The positive rate of fluorescent staining was higher（P<0.05）. In 27 superficial samples of dandruff and nail dandruff,the positive rates of fungi were 85.19%,59.26% and 55.56% by fluorescent staining,KOH wet film and fungal culturing,respectively（P<0.05）. The positive rates of fungi by fluorescent staining and ink staining in 30 cerebrospinal fluid samples had no statistical significance（P>0.05）. Conclusions Fluorescent staining is simple,rapid and specific,and it has certain clinical value in the diagnosis and identification of fungi.

A preliminary study on Lp-PLA2 and its relationship with LDL-C and hs-CRP in southern China

ZHAO Jingjing, ZHANG Peng, ZHOU Yong, CAO Yingping, HUANG Zhihu, FU Shengmiao, WANG Yuming, HONG Guolin, SHEN Jun, ZHENG Lei

2021, 36(12):  1222-1228.  DOI: 10.3969/j.issn.1673-8640.2021.012.006

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Objective To investigate the distribution of lipoprotein-associated phospholipase A2（Lp-PLA2） levels in southern China,and to evaluate the relationship between Lp-PLA2 and low-density lipoprotein cholesterol（LDL-C）,high-sensitivity C-reactive protein（hs-CRP）. Methods A total of 2 318 serum samples were collected from 8 Grade 3 Class A hospitals in southern China. Lp-PLA2,blood lipids [total cholesterol（TC）,triglyceride（TG） and high-density lipoprotein cholesterol（HDL-C）] and hs-CRP levels were determined. All the subjects were classified according to sex and ages（<50 years old,50-59 years old,60-69 years old,70-79 years old and ≥80 years old）. Pearson or Spearman correlation analysis was performed to evaluate the correlations between the items. Results The Lp-PLA2 level of 2 318 healthy subjects in southern China was（145.84±43.96） ng/mL,and the upper limit of reference interval was 217.93 ng/mL. The level of serum Lp-PLA2 in females was lower than that in maless（P<0.05）. The level of serum Lp-PLA2 in <50 years old group was lower than those in 50-59,60-69 and 70-79 years old groups（P<0.05）,and there was no statistical significance compared with that in ≥80 years old group（P>0.05）. There was no statistical significance in serum Lp-PLA2 levels among 50-59,60-69 and 70-79 years old groups（P>0.05）. Among the 8 hospitals,the level of Lp-PLA2 in the Union Hospital Affiliated to Fujian Medical University was the lowest,while the level of Lp-PLA2 in Hainan Provincial People's Hospital was the highest,and there was statistical significance compared with the other hospitals（P<0.05）. Lp-PLA2 was positively correlated with LDL-C（r=0.163,P<0.05）,and there was no correlation with TC,TG,HDL-C and hs-CRP（r values were -0.028,0.000,0.001 and 0.088,P>0.05,respectively）. Conclusions The reference interval of Lp-PLA2 in healthy subjects in southern China has been established preliminarily.

TLR4 mRNA and HO-1 level in evaluating the condition and prognosis of ICVD

FAN Minghe, LIN Guangmin, HUANG Ruijie

2021, 36(12):  1229-1233.  DOI: 10.3969/j.issn.1673-8640.2021.012.007

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Objective To investigate the roles of heme oxygenase-1（HO-1） and Toll-like receptor（TLR） 4 mRNA in evaluating the condition and prognosis of ischemic cerebrovascular disease（ICVD）. Methods Totally,324 patients with ICVD（ICVD group） and 324 healthy subjects（control group） were enrolled. HO-1 level and TLR4 mRNA expression in peripheral blood were determined. According to the National Institutes of Health Stroke Scale（NIHSS）score,ICVD patients were classified into mild injury group,moderate injury group and severe injury group. According to the results of head Doppler ultrasound,ICVD patients were classified into mild stenosis group,moderate stenosis group and severe stenosis group. These patients with ICVD were followed up for 6 months. According to the follow-up results,they were classified into good prognosis group and poor prognosis group. Pearson correlation analysis was used to evaluate the correlation between HO-1,TLR4 mRNA and NIHSS score. Spearman correlation analysis was used to evaluate the correlation between HO-1,TLR4 mRNA and the degree of intracranial artery stenosis. Receiver operating characteristic（ROC） curve was used to evaluate the prognostic value of HO-1 and TLR4 mRNA in ICVD. Results HO-1 level and TLR4 mRNA expression in ICVD group were higher than those in control group（P=0.000）. HO-1 level and TLR4 mRNA expression in mild stenosis group,moderate stenosis group,severe stenosis group and mild injury group,moderate injury group and severe injury group were increased in turn（P<0.05）. The HO-1 level and TLR4 mRNA expression in good prognosis group were lower than those in poor prognosis group（P<0.05）. In ICVD group,HO-1 level and TLR4 mRNA expression were positively correlated with stenosis degree and NIHSS score（P<0.05）. ROC curve analysis showed that the optimal cut-off values of HO-1 and TLR4 mRNA for predicting ICVD prognosis were 4.364 ng/mL and 0.989,the areas under curves were 0.800 and 0.815,the sensitivities were 81.4% and 85.1%,and the specificities were 72.4% and 73.5%,respectively. Conclusions HO-1 and TLR4 may be used as effective indicators for evaluating the condition and prognosis of ICVD patients.

Analysis of serum vitamin E level and distribution range in single pregnant women during different gestational weeks

WANG Peng, TAI Shuhong, MENG Xiangying, NIU Chao, LI Songlei, YUAN Enwu

2021, 36(12):  1234-1237.  DOI: 10.3969/j.issn.1673-8640.2021.012.008

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Objective To investigate the distribution range of serum vitamin E（Vit E） levels in single pregnant women during different gestational weeks. Methods A total of 478 non-pregnant and 3 558 single pregnant women aged 20-35 years and took progestational or gestational healthy examination were enrolled. According to gestational weeks,these pregnant women were classified into 9 groups,6-8,9-12,13-16,17-20,21-24,25-28,29-32,33-36 and 37-40 gestational week groups. Based on the method of high-performance liquid chromatography,the levels of Vit E in serum were evaluated. Spearman correlation analysis was used to evaluate the correlation between serum Vit E level and pregnancy. The percentile method [P₅₀（P_2.5-P_97.5）] was used to determine the distribution range of Vit E during different gestational weeks. Results Vit E levels in serum of non-pregnant women had no statistical significance among the age groups（P>0.05）. The difference of Vit E levels had no statistical significance among the non-pregnant,6-8,9-12 gestational week groups,and among the 29-32,33-36,37-40 gestational week groups（P>0.05）. The levels of Vit E gradually increased with the pregnancy in 13-16,17-20,21-24,25-28 gestational week groups（P<0.05,P<0.01）. By Spearman correlation analysis,the Vit E levels had a positive correlation with the gestational weeks（r=0.66,P<0.01）. Merged the gestational weeks whose Vit E levels had no statistical significance,the distribution ranges of serum Vit E level in different gestational weeks were established by percentile method：6-12 gestational weeks 10.00（6.20-17.11） mg/L,13-16 gestational weeks 11.60（7.70-18.29） mg/L,17-20 gestational weeks 12.60（8.70-18.20） mg/L,21-24 gestational weeks 14.00（9.40-21.40） mg/L,25-28 gestational weeks 15.20（9.70-22.50） mg/L and 29-40 gestational weeks 16.80（10.11-25.50） mg/L. Conclusions From 13 to 28 gestational weeks,the level of Vit E increases. The upper limit of Vit E distribution ranges in the groups of 21-40 gestational weeks is higher than that of healthy adults.

Application value of serum sST2 level in monitoring rheumatoid arthritis and therapeutic effect

ZHANG Wei, GUO Huijuan, WU Ximei, LING Xiuting, JIANG Xiaoxin, LIU Yunhong

2021, 36(12):  1238-1242.  DOI: 10.3969/j.issn.1673-8640.2021.012.009

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Objective To investigate the value of serum soluble stromelysin 2（sST2）level in the observation of rheumatoid arthritis（RA） and the monitoring of therapeutic effect. Methods Serum sST2 levels in 75 RA patients（RA group）and 50 healthy subjects（control group） were determined by enzyme-linked immunosorbent assay（ELISA）,and C-reactive protein（CRP）,erythrocyte sedimentation rate（ESR）and rheumatoid factor（RF） in RA patients were determined as well. Disease activity score 28（DAS28） was used to evaluate the disease activity of RA and drug efficiency. Of the 33 RA patients who received drug therapy,28 patients were followed up for 2 treatment cycles,and 5 patients were followed up for 4 treatment cycles. Pearson correlation analysis was used to evaluate the correlation between sST2 and various indicators. Multiple regression analysis was used to evaluate the influencing factors of serum sST2 level. Results The level of serum sST2 in RA group was higher than that in control group（P<0.01）. Pearson correlation analysis showed that serum sST2 level was positively correlated with CRP,ESR and DAS28（r values were 0.534,0.487 and 0.794,respectively,P<0.05）,while no correlation was observed between sST2 and RF（r=0.095,P>0.05）. Multiple regression analysis results showed that CRP and ESR were independent factors influencing serum sST2 level [the regression equation was sST2=28.162+0.249×CRP+0.091×ESR（r²=0.675,P<0.05）]. Serum sST2,CRP,ESR and DAS28 in RA patients were decreased after treatment（P<0.01）,and there was no statistical significance in RF before and after treatment（P>0.05）. There was a positive correlation between the change of serum sST2 level（△sST2）and the change of DAS28（△DAS28） in RA patients before and after treatment（r=0.447,P=0.002）. Conclusions The indicator,sST2,may be used to observe the change of RA and the monitoring of therapeutic effect.

Correlation between serum calbindin and mortality in hemodialysis patients with hyperphosphatemia

BAO Binan, PAN Xiaoliang, CHEN Li

2021, 36(12):  1243-1247.  DOI: 10.3969/j.issn.1673-8640.2021.012.010

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Objective To investigate the correlation between serum calbindin（CPT） and the risk of all-cause death in hemodialysis patients with hyperphosphatemia. Methods Totally,373 continuous hemodialysis patients with hyperphosphatemia were enrolled. The baseline clinical data of all the patients [age,sex,body mass index（BMI）,ankle brachial index（ABI）,smoking history,combined diseases and so on] and clinical laboratory test data [white blood cell（WBC） count,high-sensitivity C-reactive protein（hs-CRP）,platelet（PLT）,albumin（Alb）,urea nitrogen,low-density lipoprotein cholesterol（LDL-C）,triglyceride（TG）] were collected. Serum CPT was determined. All the patients were classified into low CPT group,medium CPT group and high CPT group according to CPT levels. All the patients were followed up for 4-80 months. The endpoint of follow-up was death,renal transplantation or follow-up completion. Pearson correlation analysis was used to evaluate the correlation between various indexes and CPT levels. Cox regression analysis was used to evaluate the relationship between death risk and CPT level in hemodialysis patients. Results Serum CPT level of 373 continuous hemodialysis patients with hyperphosphatemia was 6.1（4.3-8.7） ng/L. Serum CPT in males was higher than that in females（P<0.01）. There was no statistical significance in serum CPT levels whether with smoking history,diabetes mellitus history,coronary artery disease history,stroke history and peripheral arterial disease history or not（P>0.05）. The results of correlation analysis showed that CPT was negatively correlated with dialysis time（r=-0.174,P<0.001）. It was positively correlated with BMI,WBC count,hs-CRP,PLT count and TG（r values were 0.192,0.553,0.378,0.329 and 0.270,P<0.05）,and there was no correlation with Alb and LDL-C（r values were 0.092 and -0.005,P>0.05）. Kaplan-Meier curve analysis indicated that the median survival time of patients in high CPT group was shorter than those in low and medium CPT groups（P<0.001）. Compared with medium CPT group,the risk of death in low CPT group decreased by 11% [hazard ratio（HR）=0.89,95% confidence interval（CI） 0.78-0.98,P=0.03] after adjusting for age,sex,BMI,hypertension history,diabetes mellitus history,dyslipidemia,dialysis time and smoking history. The risk of death increased by 70% in high CPT group（HR=1.70,95%CI 1.52-2.33,P=0.001）. Conclusions The level of serum CPT is related to the risk of death in hemodialysis patients with hyperphosphatemia.

Relationship between plasma Nesfatin-1 and DKK-1 levels and osteonecrosis of the femoral head

PANG Shilong, WANG Han, ZHANG Zhiqiang, LI Qiang, ZHU Rusen

2021, 36(12):  1248-1252.  DOI: 10.3969/j.issn.1673-8640.2021.012.011

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Objective To investigate the relationship between plasma Nesfatin-1,Dickkopf-1（DKK-1） and osteonecrosis of the femoral head（ONFH）. Methods A total of 85 patients with ONFH（ONFH group） and 50 healthy subjects（control group） were enrolled. The levels of plasma Nesfatin-1 and DKK-1 were determined. Spearman rank correlation analysis was used to evaluate the correlation between Nesfatin-1,DKK-1 and the Association Research Circulation Osseous（ARCO）staging. Receiver operating characteristic（ROC） curve was used to evaluate the efficiency of each index in the diagnosis of femoral head collapse. Results The plasma levels of Nesfatin-1 and DKK-1 in ONFH group were higher than those in control group（P<0.05）. The plasma levels of Nesfatin-1 and DKK-1 in ONFH group increased with the increase of ARCO staging（P<0.05）. The plasma levels of Nesfatin-1 and DKK-1 in patients with femoral head collapse were higher than those without femoral head collapse（P<0.05）. Spearman rank correlation analysis results showed that plasma Nesfatin-1 and DKK-1 levels were positively correlated with ARCO staging（r_s=0.733 and 0.626,P<0.05）. ROC curve analysis showed that the areas under curves（AUC）of Nesfatin-1,DKK-1 single and combined determinations in the diagnosis of femoral head collapse were 0.707,0.643 and 0.889,respectively. Conclusions The plasma Nesfatin-1 and DKK-1 levels are related to ONFH.

Evaluation of 2 blood cultures' performance with different antibiotic adsorption materials in detecting pathogens on BacT/ALERT system

ZHENG Xin, HUANG Qi, XING Shan, TAN Xiaoli, DAI Shuqin, LIU Wanli, LIU Xiaomin

2021, 36(12):  1258-1263.  DOI: 10.3969/j.issn.1673-8640.2021.012.013

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Objective To compare and evaluate the ability to detect pathogens between blood culture bottles of new antibiotic adsorption materials and those of activated carbon adsorption materials. Methods According to the common bacterial distribution of Sun Yat-sen University Cancer Center,18 selected isolates（clinical isolates and ATCC standard isolates） were chosen to simulate patients' samples with non-antibiotic treatment and antibiotic treatment including vancomycin,daptomycin,imipenem and piperacillin-tazobactam. Those samples were added into blood culture bottles of 2 different types of antibiotic adsorption materials to evaluate their positive rates and detection times in BacT/ALERT system. Results Blood cultures of new antibiotic adsorption material and activated carbon adsorption material showed 100% recovery on 18 pathogens,like Gram-positive and Gram-negative bacteria and yeast,except for Staphylococcus haemolyticus. The time to detection of 13 species of pathogens was faster in new antibiotic adsorption material when compared with activitied carbon adsorption material（P<0.05）. The average time to detection in new antibiotic adsorption aerobic material was 0.72-68.24 h,which was faster than that in activated carbon adsorption aerobic material,while the time to detection of new antibiotic adsorption anaerobic material showed 0.48-14.48 h shorter than that of activated carbon adsorption anaerobic material in detecting pathogens. When specific antibiotic added,both new antibiotic adsorption materials showed 100% recovery to 3 pathogens,while the recovery in activated carbon adsorption materials was 33%. The average time to detection was ranged from 12.72 h to 18.08 h in new antibiotic adsorption aerobic material and from 11.76 h to 28.16 h in new antibiotic adsorption anaerobic material. Conclusions In the absence of antibiotic interference,blood cultures with different antibiotic adsorption materials show the same positive rate of target pathogen,and those of new antibiotic adsorption material have a shorter time to detection. In the presence of antibiotic interference,compared with the activated carbon adsorption material,the blood culture bottles of the new antibiotic adsorption material have a higher detection ability of pathogens.

Inhibit miR-4429 proliferation,migration and invasion of prostate cancer cells by targeting metadherin

YU Ying, LI Jianjie, LI Hanhua, HUANG Juan, WENG Wenhao, CHEN Xuefei

2021, 36(12):  1267-1273.  DOI: 10.3969/j.issn.1673-8640.2021.012.015

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Objective To study the effects of microRNA（miR）-4429 on the proliferation,migration and invasion of prostate cancer cells,and to investigate the potential related regulatory mechanism. Methods Real-time fluorescence quantitative polymerase chain reaction（RT-qPCR）was used to determine the relative expression of miR-4429 in prostate cancer cell lines（DU145,22rv1,PC3,LNCaP,VCaP） and human prostate epithelial cells（RWPE-1）. The subjects were classified into miR-4429 mimic group,miR-4429 inhibitor group,mimic negative control（NC） group and inhibitor NC group. The effects of miR-4429 on proliferation,migration and invasion of PC3 cells were determined by CCK-8,colony formation test,Transwell chamber test and matrix gel invasion test. The possible target genes of miR-4429 were predicted by bioinformatics analysis,and the binding of miR-4429 and metadherin（MTDH）was determined by luciferase reporter assay. The effects of miR-4429 on MTDH mRNA and protein levels were determined by RT-qPCR and western blotting. Results The relative expression of miR-4429 in 5 prostate cancer cell lines was lower than that in normal human prostate epithelial cells（RWPE-1）（P<0.05）,especially in PC3 cells（P<0.01）. After transfection with miR-4429 mimic,the relative expression of miR-4429 was higher than that in mimic NC group（P<0.01）,and the numbers of clone formation,migration and invasion cells were lower than those in mimic NC group（P<0.01）. After transfection with miR-4429 inhibitor,the relative expression of miR-4429 was lower than that in inhibitor NC group（P<0.01）,and the numbers of clone formation,migration and invasion cells were higher than that in inhibitor NC group（P<0.01）. The binding sites of miR-4429 and MTDH were confirmed by luciferase reporter assay. Compared with RWPE-1,the relative expressions of MTDH mRNA and protein in PC3 cells were increased（P<0.01）. The relative expressions of MTDH mRNA and protein in miR-4429 mimic group were lower than those in mimic NC group（P<0.01）. The relative expressions of MTDH mRNA and protein in PC3 cells transfected with miR-4429 inhibitor were higher than those in inhibitor NC group（P<0.01）. The proliferation,migration and invasion abilities of PC3 cells were promoted by MTDH overexpression. MTDH overexpression reversed the inhibitory effect of miR-4429 on proliferation,migration and invasion of PC3 cells. Conclusions Inhibit miR-4429 could inhibit the proliferation,migration and invasion of PC3 cells,and its mechanism is the down-regulation of MTDH.

Performance specification for differential blood count project in Shanghai

MIAO Yingbo, SONG Ying, ZHAO Qiang, ZHU Peichao, ZHOU Wei, WANG Qing, XU Chong

2021, 36(12):  1274-1276.  DOI: 10.3969/j.issn.1673-8640.2021.012.016

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Objective To find a performance specification for differential blood count project,and to improve the determination level of differential blood count project in Shanghai continuously and effectively. Methods The data of 60 batches of differential blood count project for 12 times from 2015 to 2020 were collected,and the passing rates of all the batches in each year were evaluated according to the performance specifications based on biological variation and the state-of-the-art. The performance specifications with passing rates ≥80% were selected as the performance specifications for differential blood count project. Results All the batches of neutrophil,eosinophil and basophil passed ≥80% when using the optimal performance specification（11.2%,18.6% and 19.2%） based on biological variation. All the batches had a passing rate of ≥80% for lymphocyte when using the minimum performance specification（24%） based on biological variation. All the batches of monocyte were counted using a performance specification based on the state-of-the-art（<1,±0.279;≥1,±27.9%）,and the passing rates were ≥80%. Conclusions The performance specifications for neutrophil,lymphocyte,eosinophil and basophil in Shanghai can be 11.2%,24.0%,18.6% and 19.2%,respectively,and those for monocyte<1 and monocyte≥1 are ±0.279 and ±27.9%,respectively.

Research progress and application of RNase H-dependent PCR technology

WU Xinxin, HAN Mingyue, YU Daojun

2021, 36(12):  1277-1282.  DOI: 10.3969/j.issn.1673-8640.2021.012.017

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Ribonuclease（RNase） H-dependent polymerase chain reaction（PCR）（rhPCR） adds RNase H2 and blocked cleavable rhPCR primers based on PCR,which eliminates the formation of primer-dimers in the reaction process,greatly reduces the occurrence of error amplification,and significantly improves the specificity,sensitivity and repeatability of the reaction. The rhPCR shows certain advantages in single nucleotide polymorphism（SNP）,genotyping,multi-target simultaneous detection,environmental nucleic acid detection and so on. This review summarizes the background,principles,key technology,technical advantages,application and development prospects of rhPCR.

Research progress of mitochondria in platelets

ZHAO Xiaoxia

2021, 36(12):  1283-1286.  DOI: 10.3969/j.issn.1673-8640.2021.012.018

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Platelet is nuclear-free,and mitochondria play roles in it. This review focuses on the mitochondria in energy metabolism,adenosine triphosphate（ATP） production,activation and apoptosis of platelet and the pathophysiologic role in systemic diseases,which provides a reference for further research and clinical treatment.

Research progress in the methodology and clinical application of low-density lipoprotein receptor gene mutation and function determination

ZHU Yonglin, WANG Lüya, YAN Shengkai

2021, 36(12):  1287-1291.  DOI: 10.3969/j.issn.1673-8640.2021.12.019

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Low-density lipoprotein receptor（LDLR） is one type of cell surface glycoprotein,which plays a role in maintaining cholesterol metabolism balance in vivo. LDLR gene mutation can cause the decreasing,even the absence of LDLR on the cell surface,reduce the body's ability to metabolize cholesterol,lead to elevated plasma low-density lipoprotein cholesterol（LDL-C） level and deposition in different tissues and organs,and cause a variety of lipid metabolism disorders. Accurate and rapid determination of LDLR gene mutations and analysis of changes in LDLR function are essential for the diagnosis and differential diagnosis of familial hypercholesterolemia（FH）. This review focuses on the progress in the methodology and clinical application of LDLR gene mutation and function determination,in order to raise awareness and concern of its importance in early diagnosis and intervention of FH patients.