Table of Content

30 June 2022, Volume 37 Issue 6

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Original article

Drug resistance and biofilm formation abilities of Corynebacterium striatum with different genotypes

SUN Wei, XIA Shuai, FAN Yujian, MA Liyan

2022, 37(6):  505-509.  DOI: 10.3969/j.issn.1673-8640.2022.06.001

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Objective To study the characteristics of drug susceptibility and biofilm formation ability of Corynebacterium striatum with different genotypes,and to provide a reference for clinical diagnosis and therapy. Methods Totally,36 isolates of Corynebacterium striatum were collected from January 2018 to December 2021,which included 21 isolates from sterile body fluid specimens and the other 15 isolates from clean middle urine specimens. Multilocus sequence typing（MLST） genotyping,drug susceptibility test（agar dilution method） to 10 agents and biofilm formation determination（MTT） were carried out. Results Totally,36 isolates of Corynebacterium striatum were isolated mainly from immuno-suppressed elder patients,9 genotypes were classified with MLST method,and ST6（36.1%） and ST2（25.0%） were the most popular genotypes. All the 36 isolates were multi-drug resistance（MDR） bacteria,and they were sensitive to vancomycin and linezolid（100%）. Most ST2 isolates were sensitive to tetracycline,while most ST6 isolates were sensitive to gentamicin and rifampin. The biofilm formation determination showed that all the 36 isolates could form biofilm on the polystyrene surface,but the ST2 ones had stronger biofilm formation ability compared with the other genotypes（P<0.05）. Conclusions Different genotypes of Corynebacterium striatum have different biofilm formation abilities and drug susceptibilities to commonly used antibiotics. In the process of clinical diagnosis and treatment,clinicians should pay more attention to the rational use of antibiotics,which will help to avoid opportunistic infections and nosocomial infections.

Correlation of seminal plasma SIRT1 protein expression level with ages and sperm parameters

ZHOU Fang, GU Benhong, HAN Wenjun, LI Baisheng, LE Wei, Labapingcuo , GUO Jianbing, ZHANG Jin, ZHANG Jinfu

2022, 37(6):  510-513.  DOI: 10.3969/j.issn.1673-8640.2022.06.002

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Objective To analyze the correlation of seminal plasma silent information regulator 1（SIRT1） protein expression level with sperm parameters and ages. Methods A total of 110 male patients of 21-40-year-old from the andrology departments of Tongren Hospital of Shanghai Jiao Tong University School of Medicine and Shanghai Seventh People's Hospital were enrolled,and their semen analysis and seminal plasma SIRT1 protein expression level were performed and determined. The relation was evaluated among age,sperm parameters and seminal plasma SIRT1 protein expression levels. The control group was composed of 60 patients with normal sperm parameters,and the abnormal group was consisted of 50 patients with abnormal sperm parameters. The research subjects were classified into 21-30-year-old and 31-40-year-old groups. Seminal plasma SIRT1 protein expression levels in these groups were compared. Results The expression level of seminal plasma SIRT1 protein was positively correlated with age,sperm concentration and progressive motility sperm rate（PR）（r values were 0.28,0.42 and 0.29,P<0.01）. According to different ages,seminal plasma SIRT1 protein expression level in abnormal group was lower than that in control group（P<0.01）. In 31-40-year-old group,seminal plasma SIRT1 protein expression level in abnormal group was lower than that in control group（P<0.05）. According to different sperm parameters,seminal plasma SIRT1 protein expression level in 31-40-year-old group was higher than that in 21-30-year-old group（P<0.01）,and it of 31-40-year-old patients was also higher than that of 21-30-year-old patients in control group（P<0.01）. Conclusions Seminal plasma SIRT1 protein expression is related to ages and sperm parameters,decreasing with sperm quantity and quality and increasing with age,which could be utilized to assess male reproductive potential.

Correlation of hematological indexes and expression of HER-2 in patients with breast cancer

YU Fangfang, ZHAO Qi, YANG Liping, WANG Chenyu

2022, 37(6):  514-517.  DOI: 10.3969/j.issn.1673-8640.2022.06.003

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Objective To investigate the correlation of hematological indexes and expression of human epidermal growth factor receptor-2（HER-2） in patients with breast cancer. Methods Totally,133 patients with invasive breast cancer were enrolled,and they were classified into HER-2 positive group（41 cases） and HER-2 negative group（92 cases）. The clinical characteristics and hematological indexes were compared between the 2 groups. Results Compared with HER-2 negative group,HER-2 positive group had a higher red cell volume distribution width（RDW） and a lower platelet distribution width（PDW）（P<0.05）. The optimal cut-off values of RDW and PDW for the differential diagnosis of HER-2 expression were 13.35% and 49.70 fL,respectively. The expression of HER-2 was correlated with lymph node metastasis（P<0.05）. A high level of RDW（≥13.35%） and a low level of PDW（<49.70 fL） were correlated with HER-2 expression in breast cancer tissues（P<0.05）. Conclusions RDW and PDW are correlated with HER-2 expression in breast cancer tissues,and they provide a reference for the differential diagnosis of benign and malignant breast cancer.

Clinical values of IL-18,IL-37,NLRP3 and NK/DC ratio in hepatitis B-related liver cirrhosis

WU Sujun, JI Hengtao, PENG Mengle

2022, 37(6):  518-523.  DOI: 10.3969/j.issn.1673-8640.2022.06.004

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Objective To investigate interleukin-18（IL-18）,interleukin-37（IL-37）,nucleotide-binding oligomerization domain-like receptor 3（NLRP3）and natural killer（NK）cell/dendritic cell（DC） ratio in hepatitis B-related liver cirrhosis. Methods A total of 129 patients with hepatitis B-related liver cirrhosis（hepatitis B-related liver cirrhosis group） were enrolled,and according to the ChildPugh classification,they were classified into 49 cases of grade A,47 cases of grade B and 33 cases of grade C. Totally,129 hepatitis B patients（hepatitis B group） and 129 healthy subjects（healthy control group）were enrolled as well. According to the progress of the disease,129 patients with hepatitis B-related liver cirrhosis were classified into hepatitis B-related liver cirrhosis group （98 cases）and primary liver cancer group（31 cases）. All the subjects were determined for IL-18,IL-37,NLRP3 and NK/DC ratio. Receiver operating characteristic（ROC）curve was used to evaluate the efficacy of IL-18,IL-37,NLRP3 and NK/DC ratio in the differential diagnosis of hepatitis B-related liver cirrhosis and primary liver cancer. Results The levels of IL-18,IL-37 and the relative expression of NLRP3 in healthy control group,hepatitis B group and hepatitis B-related liver cirrhosis group were increased in turn（P<0.05）,and the NK/DC ratios were decreased in turn（P<0.05）. The levels of IL-18,IL-37 and the relative expression of NLRP3 in ChildPugh grade A,B and C were increased in turn（P<0.05）,and the NK/DC ratios were decreased in turn（P<0.05）. The levels of IL-18,IL-37 and the relative expression of NLRP3 in primary liver cancer group were higher than those in hepatitis B-related liver cirrhosis group（P<0.05）,and the NK/DC ratio in primary liver cancer group was lower than that in hepatitis B-related liver cirrhosis group（P<0.05）. ROC curve analysis results showed that the areas under curves（AUC）of NLRP3,IL-18,IL-37,NK/DC ratio single determinations and the combined determination for the differential diagnosis of hepatitis B-related liver cirrhosis and primary liver cancer were 0.893,0.886,0.860,0.838 and 0.982,respectively. Conclusions IL-18,IL-37,NLRP3 and NK/DC ratio are related to the occurrence and development of hepatitis B-related liver cirrhosis,and they have clinical values in the differential diagnosis of hepatitis B-related liver cirrhosis and primary liver cancer.

Prognostic evaluation of IL-6 combined with YKL-40 in head and neck squamous cell carcinoma

WANG Xiaofei, FU Jiangtao, ZHU Shuangmei

2022, 37(6):  524-528.  DOI: 10.3969/j.issn.1673-8640.2022.06.005

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Objective To investigate the prognostic roles of interleukin（IL）-6 and cartilage glycoprotein 39（YKL-40） for evaluating all-cause mortality in patients with head and neck squamous cell carcinoma（HNSCC）. Methods The data from 130 patients with HNSCC were analyzed. Serum IL-6 and YKL-40 were determined. All the patients were followed up,and the primary endpoint was all-cause mortality. Receiver operating characteristic （ROC） curve was used to determine the optimal cut-off values for the prognosis of HNSCC patients by IL-6 and YKL-40. Kaplan-Meier curve was used to analyze the survival of HNSCC patients. Cox proportional hazard model was used to determine risk factors for overall survival in patients with HNSCC. Results The optimal cut-off values of IL-6 and YKL-40 were 19.8 pg/mL and 119 μg/L. Compared with IL-6≤19.8 pg/mL,patients in IL-6>19.8 pg/mL had increased proportions of stage Ⅲ and Ⅳ in TNM classification（P<0.05）. There was no statistical significance for sex,age,tumor location and treatment method between the 2 groups （P>0.05）. Increased age and proportions of stage Ⅲ and Ⅳ were also noticed in YKL-40>119 μg/L group as compared with those in YKL-40≤119 μg/L group（P<0.05）. There was no statistical significance in sex,tumor location and treatment method between the 2 groups （P>0.05）. Survival analysis indicated that patients in IL-6>19.8 pg/mL group and YKL-40>119 μg/L group had reduced median survival time as compared with those in IL-6≤19.8 pg/mL group（P<0.001） and YKL-40≤119 μg/L group（P=0.002）. Cox regression analysis showed that TNM classification [hazard ratio（HR）=2.65,95% confidence interval（CI） 1.64-5.64],IL-6（HR=1.78,95%CI 1.43-3.07） and YKL-40（HR=1.69,95%CI 1.28-2.10）were risk factors for overall survival in patients with HNSCC. Conclusions IL-6 and YKL-40 are related to the prognosis of HNSCC patients,and the combined determination can be used in evaluating patients' prognosis.

Relationship between ADAMTS-13 activity and non-ST-segment elevation myocardial infarction

WANG Bin

2022, 37(6):  529-534.  DOI: 10.3969/j.issn.1673-8640.2022.06.006

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Objective To investigate the clinical role of a disintegrin and metalloproteinase with thrombospondin motif 13（ADAMTS-13）in identifying high-risk non-ST-segment elevation myocardial infarction（NSTEMI）. Methods ADAMTS-13 activity determination was performed on the samples collected from 142 patients with NSTEMI（NSTEMI group） through a fluorescence resonance energy transfer（FRET）-based assay. Totally,50 non-coronary heart disease patients with normal coronary angiography were enrolled as control group during the same period. The baseline information,clinical data and determination results [creatinine（Cr）,high-sensitivity cardiac troponin I（hs-cTnI）,platelet（PLT） count,D-dimer（DD）] were collected. According to the Global Registry of Acute Coronary Events（GRACE）score at admission,the NSTEMI patients were classified into low-risk group（43 cases）,medium-risk group（54 cases）and high-risk group（45 cases）. Spearman rank correlation analysis was used to analyze the correlation between ADAMTS-13 activity and risk stratification in NSTEMI patients. Binary Logistic regression analysis was used to investigate the influencing factors for high-risk NSTEMI. Receiver operating characteristic（ROC）curve was used to evaluate the clinical value of ADAMTS-13 on high-risk NSTEMI. Results The ADAMTS-13 activity in NSTEMI group was lower than that in control group（P<0.001）,and it was decreased from low-risk,medium-risk to high-risk groups（P<0.01）. Spearman rank correlation analysis results showed that,ADAMTS-13 activity was negatively correlated with risk stratification in NSTEMI patients（r_s=-0.527,P<0.001 ）. Binary Logistic regression analysis showed that ADAMTS-13 was a protective factor [odds ratio（OR）=0.936,95% confidence interval（CI） 0.898-0.975]. Age,heart rate,Cr and Killip grading ≥Ⅱ were risk factors（OR=1.148,1.085,1.023 and 26.207,95%CI 1.047-1.259,1.023-1.150,1.004-1.041 and 5.489-125.136）. ROC curve analysis indicated that the area under curve（AUC） of ADAMTS-13 activity in predicting high-risk NSTEMI was 0.784. When the optimal cut-off value was 61.7%,the sensitivity and specificity were 71.1% and 81.4%,respectively. Conclusions ADAMTS-13 activity is related to the risk stratification of NSTEMI,which has clinical value in the diagnosis of high-risk NSTEMI.

Clinical role of blood lipid in brittle fracture elderly patients with osteoporosis

SHEN Jianguo, LI Tingting, SONG Yunxiao, BIAN Xiaobo, ZHANG Linlin, JIN Xiaoling

2022, 37(6):  535-538.  DOI: 10.3969/j.issn.1673-8640.2022.06.007

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Objective To investigate the clinical diagnosis and predictive value of blood lipid index in brittle fracture elderly patients with osteoporosis. Methods Totally,196 patients with osteoporosis and 129 patients with brittle fracture were enrolled. Low-density lipoprotein cholesterol（LDL-C）,triglyceride（TG）,small and dense low-density lipoprotein cholesterol（sd-LDL-C）,high-density lipoprotein cholesterol（HDL-C）,apolipoprotein（apo） A,apo B,apo E,total cholesterol（TC） and lipoprotein a [Lp（a）] were determined. Binary Logistic regression analysis was used to adjust for confounding factors to evaluate the risk factors of brittle fracture in elderly patients with osteoporosis. Results The levels of LDL-C,TG,TC and apo A in brittle fracture group were higher than those in osteoporosis group,while the levels of HDL-C and sd-LDL-C were lower than those in osteoporosis group（P<0.05）. There was no statistical significance in apo B,apo E and Lp（a）levels between the 2 groups（P>0.05）. Multivariate binary Logistic regression analysis showed that increased LDL-C,TG,TC,apo A and decreased HDL-C were risk factors for brittle fracture in osteoporosis patients [odds ratios（OR） were 1.473,1.543,1.745,1.535 and 0.177,95% confidence intervals（CI） were 1.094-1.985,1.125-2.114,1.381-2.206,1.098-1.848 and 0.090-0.349,respectively]. Conclusions Dyslipidemia is related with brittle fracture in elderly patients with osteoporosis.

Application of whole exome sequencing in 9p deletion syndrome

YI Wei, PAN Yunhua, LIU Houchang, YU Chongfei, YANG Biqing, GE Shijun, LIN Keqin, CHU Jiayou, YANG Zhaoqing

2022, 37(6):  539-542.  DOI: 10.3969/j.issn.1673-8640.2022.06.008

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Objective To report the clinical and molecular cytogenetic characteristics of a XX female patient with 9p deletion syndrome. Methods Chromosome G-banding karyotyping was performed on the patient and her parents,and whole exome sequencing was performed on the patient. The clinical,molecular cytogenetic characteristics and distal deletions on chromosome 9p were analyzed. Results The patient presented with primary amenorrhea,infantile uterus,abnormal level of sex hormones and epilepsy,but without typical clinical features of 9p deletion syndrome such as trigonocephaly and mental retardation. Karyotyping analysis of the patient showed 46,XX,del（9）（p23）,and her parents' karyotypes were normal. Whole exome sequencing showed the patient had a deletion on 9p24.3-p23（24 846-13 022 661,13.0 Mb）,which included critical genes such as DMRT1,DMRT3,DMRT2,DOCK8,KANK1,FOXD4（totally 61 genes）. No other pathogenic copy number variation（CNV）,base insertion and deletion were detected in other regions of chromosomes. Conclusions Haploinsufficiency of one or more genes on the terminal of chromosome 9p may underlie the disorders of sexual development of the patient. Whole exome sequencing provides genotypic data relevant to the clinical phenotypes.

Correlation of MTHFR gene C677T polymorphism and blood lipid levels with GDM

WANG Lingling, REN Chunli

2022, 37(6):  543-546.  DOI: 10.3969/j.issn.1673-8640.2022.06.009

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Objective To investigate the relationship between the polymorphism of methylene tetrahydrofolate reductase（MTHFR） gene C677T polymorphism,blood lipid levels and gestational diabetes mellitus（GDM）. Methods Totally,144 patients with GDM and 142 healthy pregnant women were enrolled as case group and control group,respectively. The polymorphism of MTHFR gene C677T and blood lipid levels [total cholesterol（TC）,triglyceride（TG）,low-density lipoprotein cholesterol（LDL-C）,high-density lipoprotein cholesterol（HDL-C）] were determined,and non- high-density lipoprotein cholesterol（non-HDL-C） levels were calculated. Results There was statistical significance in genotype distribution and allele frequency of MTHFR gene in GDM group compared with control group（P<0.01）. Compared with control group,the levels of TG,TC,LDL-C,non-HLD-C in GDM group were increased（P<0.05）,and the HDL-C level was decreased（P<0.01）. Compared with CC genotype GDM patients,TT genotype GDM patients had higher TC and non-HDL-C levels（P<0.05） and lower HDL-C levels（P<0.01）. The levels of TG,TC,LDL-C and non-HDL-C in GDM patients with CT genotype were increased（P<0.05）,while HDL-C was decreased（P<0.01）. Compared with CC genotype,TG,TC and non-HDL-C levels were increased in healthy pregnant women with TT and CT genotype. T allele frequency,LDL-C levels and non-HDL-C levels were positively correlated with GDM [odds ratio（OR）=5.25,95% confidence interval 3.66-7.53;OR=2.44,95% confidence interval 1.69-3.53;OR=1.65,95%confidence interval 1.41-1.92]. HDL-C was negatively correlated with GDM（OR=0.05,95% confidence interval 0.02-0.13）. Conclusions The polymorphism of MTHFR gene C677T may be related tso the abnormality of blood lipid in GDM,and increased MTHFR T allele frequency may be a risk factor for GDM.

Feasibility study of automated hematology analyzer on routine cell counts of serous cavity effusion

WANG Chong, ZHU Jie, YANG Shuo, GU Meixiu, PAN Baishen, WANG Beili, GUO Wei

2022, 37(6):  551-556.  DOI: 10.3969/j.issn.1673-8640.2022.06.011

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Objective To investigate the feasibility of replacing manual counting method with microscope in cell counts and white blood cell classification by automated hematology analyzer in body fluid mode. Methods A total of 287 serous cavity effusion specimens from inpatients were collected,and the manual counting method with microscope（manual method） and the instrumental determination method with automated hematology analyzer（instrumental method） were used for red blood cell and white blood cell counts and white blood cell classification. The manual method was used as the gold standard. The accuracy of instrumental method was evaluated. Results For the specimens whose white blood cell count 201-15 000 cells/μL or the red blood cell count ≥200 cells/μL,the instrumental method and manual method had good correlation in cell counts,and the 2 methods had a good correlation in white blood cell classification（P<0.01）. Conclusions The automated hematology analyzer has the advantages of rapidity,accuracy and reproducibility in cell counts and white blood cell classification of serous cavity effusion specimens. When the red blood cell and white blood cell counts are within certain linear ranges,instrumental method can replace the manual counting method with microscope.

sRapid determination of sex chromosome abnormality using double relative quantitative SD-qPCR

SUN Lei, LONG Ju, FAN Zuqian, WU Suping, LIN Guixian, LING Xiuming

2022, 37(6):  557-560.  DOI: 10.3969/j.issn.1673-8640.2022.06.012

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Objective To establish a single tube fluorescent double relative segmental duplication-quantitative polymerase chain reaction（SD-qPCR） for determining the number of chromosome X and Y simultaneously. Methods The established SD-qPCR was used for the determination of 197 amniotic fluid samples. The numbers of chromosome X and Y were determined by the relative quantitative 2^-ΔΔCt value between segmental duplications. The results are compared with chromosome karyotype analysis. Results Two pairs of segmental duplication molecular markers were screened and located on chromosome 16 and chromosome X,chromosome 1 and chromosome Y,respectively. Among the 197 samples,there were 185 samples with normal sex chromosome,5 cases of 45,X,3 cases of 47,XXX,2 cases of 47,XXY and 2 cases of 47,XYY. The results of SD-qPCR were consistent with those of chromosome karyotype analysis. Conclusions The established SD-qPCR can determine the numbers of chromosome X and chromosome Y in a single tube simultaneously. It has the advantages of multiple loci and stability,and it can be widely used in clinic.

Establishment and clinical application evaluation of plasma cfDNA determination in colorectal cancer patients

ZHU Yunjie, MA Zhengyao, SHEN Minna, ZHOU Yan, HUANG Fei, CHEN Xinning, ZHANG Chunyan, WANG Beili, GUO Wei

2022, 37(6):  561-567.  DOI: 10.3969/j.issn.1673-8640.2022.06.013

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Objective To establish and evaluate the performance and clinical application of plasma cfDNA determination in colorectal cancer patients based on next generation sequencing（NGS） platform. Methods Plasma circulating free DNA（cfDNA） determination system（cfDNA panel） for colorectal cancer patients had been established. The accuracy,precision,minimum sample amount and input of extraction,sensitivity and specificity of cfDNA panel were evaluated. The results were compared with both the results of Miseq Dx NGS platform and amplification refractory mutation system（ARMS）. Results The accuracy of cfDNA panel for colorectal cancer was 99.97%,and the determination rate of expected mutation was 100%. The minimum amount of plasma extraction was 1 mL,and the minimum amount of cfDNA input was 10 ng.The sensitivity was 0.2%. The 10 000 mg/L hemoglobin,500 mg/L bilirubin or 2% fat emulsion had no effet on the extraction of colorectal cancer cfDNA panel. Compared with the results of Miseq Dx NGS platform and ARMS,the consistency rates were 94.12% and 90.91%,respectively,and the positive and negative predictive rates of KRAS,NRAS,BRAF and PIK3CA were 100% and 99.60%,respectively. Conclusions The performance of cfDNA panel in colorectal cancer meets the needs of clinical application. The results of plasma cfDNA determination are highly consistent with the determination results of matched tumor tissue,which is a useful supplement for tissue biopsy.

Rapid determination of minimum inhibitory concentration and cut-off concentration of Mycobacterium tuberculosis by nitrate reductase assay using liquid medium

DU Jinghui, HUANG Zikun, LIU Min, GONG Jingjing, WANG Dan, LIU Jibo, LIU Xu

2022, 37(6):  568-576.  DOI: 10.3969/j.issn.1673-8640.2022.06.014

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Objective Through performing a multicenter study of nitrate reductase assay（NRA） using liquid medium for the determination of multidrug-resistant Mycobacterium tuberculosis（MDR-MTB） and extensively drug-resistant Mycobacterium tuberculosis（XDR-MTB）,to evaluate the performance of this method,and to identify the minimum inhibitory concentration（MIC） and cut-off concentrations of rifampicin,isoniazid,ofloxacin,amikacin,kanamycin and capreomycin. Methods The study was divided into 3 phases. In the Phase Ⅰ,the MIC of 6 drugs was determined. Phase Ⅱ established the cut-off concentrations of the 6 drugs. In the Phase Ⅲ,the cut-off concentrations for the 6 drugs used in NRA were verified by large numbers of clinical samples [730 isolates of Mycobacterium tuberculosis（MTB）] with agar proportion method（APM） or MGIT 960 system as reference method. The consistency and accuracy of the validation results from 4 participating laboratories were compared. Results The MIC for the 6 drugs were as follows：rifampicin 1 mg/L,isoniazid 0.2 mg/L,ofloxacin 2 mg/L,amikacin 4 mg/L,kanamycin 5 mg/L,capreomycin 5 mg/L. The cut-off concentrations for the 6 drugs were as follows：rifampicin 1 mg/L,isoniazid 0.2 mg/L,ofloxacin 2 mg/L,amikacin 2 mg/L,kanamycin 5 mg/L,capreomycin 2.5 mg/L. The accuracies for rifampicin,isoniazid,ofloxacin,amikacin,kanamycin and capreomycin were 96.4%-100.0%. The median determination time of NRA was 7 d. Conclusions NRA performed in liquid medium can provide a quick,economic and convenient method for the first-line and second-line drug susceptibility testing of MTB resource-limited settings.

Feasibility of the determination of heterogeneous drug resistance to rifampicin in Mycobacterium tuberculosis by flow cytometry

LI Hua, WANG Weiliang, XIE Bei, YANG Yu, MENG Fanrong, WANG Nan, LIU Zhihui, ZHANG Yanbin

2022, 37(6):  577-582.  DOI: 10.3969/j.issn.1673-8640.2022.06.015

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Objective To evaluate the feasibility of flow cytometry（FCM） in determining the heterogeneous drug resistance of Mycobacterium tuberculosis（MTB） to rifampicin（RFP）. Methods MTB clinical isolates were isolated by fluorescein diacetate（FDA） after treating with RFP of 12.5,25,50 and 100 μg/mL for 3,7 and 10 d,respectively. The mean fluorescence intensity（MFI） of MTB clinical isolates was determined by FCM,and the sensitivity index（MFI of treated/MFI of control） was calculated. The concentration and time of RFP treatment were chosen according to significance test of difference,which inhibited the growth of MTB clinical isolates sensitive to RFP but not affected the growth of MTB clinical isolates resistant to RFP. A total of 30 isolates of RFP sensitive clinical MTB isolates and 30 isolates of RFP resistant clinical MTB isolates were mixed to obtain hetero-resistant MTB,in which the proportions of resistant bacteria were 0%,25%,50%,75% and 100%,respectively. Hetero-resistant MTB were treated according to the above optimal RFP treatment concentration and time. After stained by FDA,the sensitivity index of samples was calculated. According to receiver operating characteristic（ROC） curve,the sensitivity index of hetero-resistant MTB with different proportions of RFP resistant bacteria and the dsetermination performance of the method were investigated. Results The optimal RFP treatment concentration and time of FDA based FCM in the determination of MTB heterogeneous drug resistance to RFP were 50 μg/mL and 10 d,respectively. To identify hetero-resistant MTB samples which containing 0%,25% and 100% RFP resistant MTB,the samples were treated with 50 μg/mL RFP for 10 d,the cut-off values of this method were 0.48 and 0.83,the sensitivities were 96.2% and 86.7%,and the specificities were 96.7% and 83.3%,respectively. When treated for 14 d,the cut-off values of this method were 0.37 and 0.77,the sensitivities of this method were 96.7% and 96.7%,and the specificities of this method were 93.3% and 86.7%,respectively. Conclusions FCM has a broad application prospect in the determination of MTB heterogeneous resistance.

Regulation of miR-338-3p on proliferation,invasion,migration and epithelial-mesenchymal transformation of thyroid papillary carcinoma cell line TPC-1

LI Tao, LI Jia, PAN Jing, LIANG Xiao, YU Lina

2022, 37(6):  583-589.  DOI: 10.3969/j.issn.1673-8640.2022.06.016

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Objective To investigate the effect of microRNA（miR）-338-3p overexpression of sirtuin （SIRT）6 on proliferation,invasion,migration and epithelial-mesenchymal transformation（EMT） of human thyroid papillary carcinoma cell line TPC-1. Methods Luciferase reporting assay was used to detect the targeting relationship between miR-338-3p and SIRT6. SIRT6 pcDNA vector was constructed to overexpress SIRT6,and miR-338-3p mimic and pcDNA-SIRT6 were transfected into TPC-1 cells either alone or in combination. The transfected plasmids were classified into 6 groups,including control group（no treatment）,mimic-NC group（transfected mimic-NC） and miR-338-3p mimic group（transfected miR-338-3p mimic）,pcDNA-SIRT6 group（transfected pcDNA-SIRT6）,mimic-NC+pcDNA-SIRT6 group（co-transfected mimic-NC and pcDNA-SIRT6） and miR-338-3p mimic+pcDNA-SIRT6 group（co-transfected with miR-338-3p mimic and pcDNA-SIRT6）. The clone genesis rate,invasive cell number,scratch healing rate of TPC-1 cells and the expression levels of E-cadherin（E-cad）,N-cadherin（N-cad） and Vimentin in epithelial cells were determined. Results The target gene of miR-338-3p was SIRT6. Compared with the control group,clone genesis rate,invasive cell number and scratch healing rate in miR-338-3p mimic group were decreased（P<0.05）. The protein expression of E-cad was increased（P<0.05）,while the protein expressions of N-cad and Vimentin were decreased（P<0.05）. The clone genesis rate,invasive cell number and scratch healing rate were increased in pcDNA+SIRT6 group. The protein expression level of E-cad was decreased（P<0.05）,while the protein expressions of N-cad and Vimentin were increased（P<0.05）. Compared with pcDNA-SIRT6 group,clone genesis rate,invasive cell number and scratch healing rate in miR-338-3p mimic+pcDNA-SIRT6 group were decreased（P<0.05）,while E-cad protein expression was increased（P<0.05）. The protein expressions of N-cad and Vimentin were decreased（P<0.05）. Conclusions The miR-338-3p overexpression targets SIRT6 to inhibit TPC-1 cell proliferation,invasion,migration and EMT.

Establishment and application of automatic review rules for clinical biochemical quantitative detection items

CAI Yongmei, WANG Haiying, MEI Yanfang, WANG Zhiwei

2022, 37(6):  590-595.  DOI: 10.3969/j.issn.1673-8640.2022.06.017

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Objective Based on the characteristics of patient population into clinical laboratories,to establish the automatic review rules of clinical biochemical items,and to ensure effective and stable passing rate of automatic review. Methods According to accreditation criteria for quality and competence of medical laboratories and autoverification of medical laboratory results for specific disciplines,personalized automatic review rules for clinical biochemical quantitative detection items were established and verified. Statistical analysis was used for the rate of automatic review without outdated normal range severity（NS） and delta check severity（DS） rules. Results Totally,40 clinical biochemical quantitative detection items had been established,and more than 400 effective rules were set. The failure to pass the automatic examination results of each item was mainly DS and NS rules,accounting for about 95%. After optimizing DS and NS rules,the pass rate of automatic examination were increased from 56.33% to 66.13%. Conclusions Totally,40 clinical biochemical quantitative detection items have been established. The NS and DS rules are the main rules for the stability of the automatic review and the accuracy and reliability of the automatic review results,which should be scientific,feasible and appropriate.

Problems and solutions on autoverification of coagulation test

FAN Qingkun, LI Ling, DU Jia, YANG Jun, ZHANG Zhenlu

2022, 37(6):  596-600.  DOI: 10.3969/j.issn.1673-8640.2022.06.018

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The autoverification of reports is the future development trend of clinical laboratories,which can significantly shorten the laboratory sample turn-around time and improve work efficiency. The autoverification of coagulation tests is less used in clinical laboratories,due to many problems such as the identification of unqualified samples and the complex rules of autoverification. The basis for the implementation of autoverification is strict and standardized quality control of pre-analysis and interconnection of information systems. Based on the premise of accurate results,clinical laboratories can set appropriate autoverification rules according to its own conditions,continuously optimize the autoverification rules for coagulation test,and gradually increase the autoverification pass rate of clinical laboratory test.