Table of Content

30 September 2023, Volume 38 Issue 9

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Clinical application and challenges of liquid biopsy biomarkers in non-small cell lung cancer

ZHOU Yunlan, SHEN Lisong

2023, 38(9):  807-811.  DOI: 10.3969/j.issn.1673-8640.2023.09.001

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Lung cancer is a malignant tumor with the highest incidence and mortality in China. Liquid biopsy offers advantages such as speed，non-invasiveness and repeatability over traditional tissue biopsy，making it of value in precision medicine and treatment decision for non-small cell lung cancer（NSCLC）. This review provides a brief overview of liquid biopsy in screening，early diagnosis，treatment and prognosis evaluation and resistance surveillance. In the future，new technology of liquid biopsy needs to be further improved for sensitivity and specificity，and standardized processes and reference intervals need to be established. Prospective clinical trials also need to be conducted before its clinical transformation.

MiR-374 promoting proliferation and invasion of breast cancer cells by targeting and down-regulating TRIM35

WANG Rong, XING Lianxiang, HUANG Keliang, LI Xin

2023, 38(9):  812-817.  DOI: 10.3969/j.issn.1673-8640.2023.09.002

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Objective To investigate the role of miR-374 in breast cancer and the relationship between miR-374 and tripartite motif-containing protein 35（TRIM35）. Methods A total of 42 patients with breast cancer were enrolled，and cancer tissues and adjacent tissues（>2 cm away from tumor resection margin） were collected，and the data of patients were collected as well. The expression of miR-374 and TRIM35 were determined in cancer tissues and adjacent tissues. Breast cancer cell line MCF-7 was classified into 3 groups according to different treatment methods，including control group（without any treatment），interference group（adding 100 nmol/L miR-374 inhibitor） and mock-vehicle group（adding 100 nmol/L negative control sequence）. The effects of miR-374 on the proliferative activity and invasiveness of breast cancer cell line MCF-7 were evaluated by cell proliferation assay and Transwell assay. Dual luciferase reporter gene assay was used to determine the interaction between miR-374 and TRIM35. Results The expression of miR-374 in cancer tissues was higher than that in adjacent tissues（P<0.001）. TRIM35 relative expression in cancer tissues was lower than that in adjacent tissues（P<0.001）. The miR-374 relative expression had statistical significance with different breast tumor sizes，distant metastasis and lymphatic metastasis（P<0.05）. There was no statistical significance for miR-374 relative expression with different ages and TNM stages（P>0.05）. The relative expression of miR-374 in interference group was lower than those in control group and mock-vehicle group（P<0.001）. The cell proliferation rate and the number of invasive cells in interference group were lower than those in mock-vehicle group and control group（P<0.001）. TRIM35 relative expressions were higher than those in mock-vehicle group and control group（P<0.001）. Dual luciferase reporter gene assay confirmed that miR-374 directly inhibited the transcriptional expression of target gene TRIM35. Conclusions MiR-374 interacts with TRIM35，and miR-374 promotes the proliferation and invasion of breast cancer cells by targeting and down-regulating the expression of TRIM35.

Transcription factors in prostate cancer progression

SUN Chuanyu, ZHAO Xiaojun, GE Shengyang, ZHANG Yang

2023, 38(9):  818-824.  DOI: 10.3969/j.issn.1673-8640.2023.09.003

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Objective To investigate the 46 differentially expressed proteins of upstream transcriptional regulation mechanism in prostate cancer. Methods GeneGo MetaCore software and IPA software were used to conduct bioinformatics analysis on differential proteins and clarify the transcriptional regulatory network of these differential proteins. GEPIA2 database was used to analyze the expression level of identified transcription factors and their impacts on the survival of prostate cancer patients. Results Totally，20 transcriptional regulatory networks were identified. According to the enrichment of differentially expressed proteins，the top 6 networks were screened out to be regulated by 6 transcription factors，specificity protein 1（SP1），p53，Yin-Yang 1（YY1），androgen receptor（AR），c-Myc and Slug. Kaplan-Meier survival curve analysis showed that prostate cancer patients with high expression of p53 had long disease-free survival and overall survival compared to patients with low expression of p53（P=0.047 and 0.019，respectively）. The disease-free survival of prostate cancer patients with low expression of AR was longer than that of patients with high expression of AR（P=0.023）. The other 4 transcription factors had no impact on the prognosis of prostate cancer patients（P>0.05）. IPA software analysis results showed that SP1，p53，YY1，AR and c-Myc were regulated by epigallocatechin gallate（EGCG），protein kinase B（AKT）1，fibroblast growth factor 2（FGF2） and beta-estradiol regulation. Conclusions By analyzing the transcriptional regulatory network of 46 prostate cancer differentially expressed proteins，6 transcription factors and 4 upstream regulatory molecules related to prostate cancer have been found，which can provide new targets for early screening and prognostic evaluation of prostate cancer.

Screening potential prognostic biomarkers of colorectal cancer based on weighted gene co-expression network analysis

LI Liangshan, ZHAO Hu, WANG Shiwen

2023, 38(9):  825-832.  DOI: 10.3969/j.issn.1673-8640.2023.09.004

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Objective To investigate hub genes related to colorectal cancer（CRC） as potential prognostic biomarkers by bioinformatics analysis. Methods The GSE33113 dataset related to CRC were obtained from Gene Expression Omnibus（GEO） database. Differentially expressed genes from CRC tissues and adjacent tissues were obtained using limma program package in R software，and Gene Ontology（GO） and Kyoto Encyclopedia of Genes and Genomes（KEGG） enrichment analysis of differentially expressed genes were performed with the DAVID online tool. Weighted gene co-expression network analysis（WGCNA） was used to output key module genes，and the intersection of key module genes and differentially expressed genes was taken as candidate hub genes. GEPIA database was used to verify the expression and prognostic value of hub genes in CRC. The diagnostic value of the hub genes for CRC was evaluated by receiver operating characteristic（ROC） curve. Gene set enrichment analysis（GSEA） was performed to explore the biological significance of hub genes in CRC. Human normal intestinal epithelial cell HIEC cell line，CRC cell line，HCT116 and HCT15 were used to deterimine the expression of hub genes by real-time quantitative polymerase chain reaction（RT-qPCR） and western blotting. Results GSE33113 dataset contains 90 CRC tissues and 6 adjacent tissues. A total of 1 211 differentially expressed genes were screened，including 505 up-regulated cases and 706 down-regulated cases，which were mostly distributed in extracellular space and extracellular region，regulating chemokine mediated signaling pathways，inflammatory response，cell division and so on. WGCNA analysis revealed 24 and 96 key module genes were most significantly correlated with clinical features，and 24 and 62 candidate hub genes were obtained，respectively. A total of 6 hub genes（AQP8，PBK，EXO1，CCNB1，DEPDC1B and KPNA2） were acquired after screening in GEPIA database. EXO1 was selected for validation，and its expression was up-regulated in CRC tissues of different datasets. GEPIA database analysis results showed that the expression level of EXO1 mRNA was high in CRC tissues，the expression level of EXO1 in CRC patients was correlated with overall survival（OS），and ROC curve analysis showed that EXO1 had high diagnostic value for CRC. The results of GSEA analysis suggested that EXO1 may regulate CRC by affecting cell cycle and DNA replication. RT-qPCR and western blotting results revealed that the expression of EXO1 was up-regulated in CRC cells. Conclusions Totally，6 hub genes related to CRC have been identified by WGCNA，among which EXO1 may serve as a potential prognostic biomarker for CRC.

Construction and validation of a model for pyroptosis-related features in lung adenocarcinoma

WANG Yangshuyi, YE WEI, LIN Zhihao, HUANG Yuenuo, FANG Tao, DONG Xiaoting

2023, 38(9):  833-841.  DOI: 10.3969/j.issn.1673-8640.2023.09.005

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Objective To construct a prognostic model of lung adenocarcinoma（LUAD）-pyroptosis genes based on bioinformatics，and to investigate the relationship between LUAD and cell pyroptosis. Methods The transcriptomic data of LUAD patients（TCGA cohort） and clinical data were downloaded from the Cancer Genome Atlas（TCGA），and the limma program package was used to identify differentially expressed pyroptosis genes. Cluster typing was obtained from TCGA cohort based on the expression characteristics of differentially expressed pyroptosis genes，and differentially expressed pyroptosis genes between clustering were identified. Using least absolute shrinkage and selection operator（LASSO）-Cox regression method，a prognostic model based on inter-clustering differentially expressed pyroptosis genes was developed，and the risk scores were calculated. TCGA cohort LUAD patients were classified into high and low risk groups using the median risk score. The clinical prognostic role of the prognostic model was analyzed using Kaplan-Meier survival curve and Cox regression analysis in conjunction with clinical characteristics. The LUAD patients from the Gene Expression Omnibus（GEO） were classified into 2 risk groups（high and low） using the median risk score of the TCGA cohort，and the clinical prognostic validation was performed. The signaling pathways and biological functions of differentially expressed pyroptosis genes in TCGA cohort were analyzed using Gene Ontology（GO） and Kyoto Encyclopedia of Genes and Genomes（KEGG） analysis. Single sample gene set enrichment analysis（ssGSEA） and CIBERSORT were used to compare immune cells and immune-related pathways. Results Totally，41 differentially expressed pyroptosis genes were identified in LUAD tissues and paracancer tissues. The results of cluster analysis showed that TCGA cohort samples could be classified into 2 cluster types，and the survival time of type 2 was longer than that of type 1（P<0.05）. Totally，11 differentially expressed pyroptosis genes（ANO1，PKHD1L1，FMO2，TDRD1，TBX4，ABCC12，AQP4，CPXM2，WFIKKN2，ATP1A4 and IGSF10） associated with survival in LUAD patients were identified. Kaplan-Meier survival curve analysis showed that the survival time of TCGA cohort high risk group was shorter than that of low risk group（P<0.05）. The same results were obtained using GEO cohort. Cox regression analysis showed that the risk score derived from the prognostic risk model was an independent risk factor for the prognosis of LUAD patients. Heat map analysis of 11 differentially expressed pyroptosis genes in the TCGA cohort showed that there was statistical significance in sex，clinical stage，T stage and N stage between high risk and low risk group（P<0.05）. The differentially expressed pyroptosis genes between high risk and low risk groups in TCGA cohort were related to chemokines mediating tumor-related signaling pathways，immune response and cell membrane function. Conclusions Pyroptosis genes play roles in tumor immunity，and the constructed 11 pyroptosis genes-related prognostic model can be used to predict the prognosis of LUAD.

Expression of RRBP1 in osteosarcoma tissues and its effect on cell biological characteristics

YANG Yuqiang, QUAN Xiaoli, WANG Liuyu, XIAN Wenfeng, YANG Hong

2023, 38(9):  842-848.  DOI: 10.3969/j.issn.1673-8640.2023.09.006

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Objective To investigate the expression of ribosome-binding protein 1（RRBP1） in osteosarcoma tissues and the effect on the cell biological characteristics of osteosarcoma U-2OS cells. Methods A total of 91 patients with osteosarcoma undergoing surgical treatment in Nanyang Second People's Hospital and the First Affiliated Hospital of Zhengzhou University from June 2014 to June 2020 were enrolled，and their osteosarcoma tissues were collected. Totally，45 patients who underwent orthopedic surgery and excluded bone tumors were enrolled，and the normal bone tissues were collected. The expressions of RRBP1 proteins in osteosarcoma tissues and normal bone tissues were determined by immunohistochemical method. The patients with osteosarcoma were followed up after surgical treatment，with a follow-up until December 2021，and the survival was recorded. The osteosarcoma cell line U-2OS was cultured and classified into si-RRBP1 group（transfected with RRBP1 interference sequence），si-NC group（transfected with negative control sequence） and blank group（only added with transfection reagent）. Normal cultured U-2OS cells were used as control group. The expressions of RRBP1 mRNA and protein in each group were determined，and the proliferative activity，migration and invasion ability of cells were determined. Results The positive expression rate of RRBP1 protein in osteosarcoma tissues was higher than that in normal bone tissues（P<0.001）. The positive rates of RRBP1 protein in osteosarcoma patients with Enneking stage ⅡB to Ⅲ and distant metastasis were higher than those in patients with Enneking stage ⅡA and without distant metastasis（P<0.05）. There was no statistical significance in the positive rates of RRBP1 protein among osteosarcoma patients with different sex，ages，tumor locations，histological types and tumor sizes（P>0.05）. The survival rate and time of patients in RRBP1 positive group were lower than those in RRBP1 negative group（P=0.003）. The relative expressions of RRBP1 mRNA and protein in si-RRBP1 group is cell were lower than those in si-NC groups，blank groups and U-2OS （P<0.05）. Compared with si-NC group and blank group，the absorbance（A） values in si-RRBP1 group at 24，48，72 and 96 h were decreased（P<0.05）. The number of migrating cells and invasive cells in si-RRBP1 group were lower than those in si-NC group and blank group（P<0.05）. Conclusions The expression of RRBP1 protein in osteosarcoma tissues is highly expressed，and it is related to patient prognosis. Down-regulated RRBP1 gene expression in osteosarcoma U-2OS cells can inhibit cell proliferative activity and inhibit cell migration and invasion.

Serum inflammatory factors combined with PSA and f-PSA in the auxiliary diagnosis of prostate cancer

PENG Wei, LI Yungai, XU Jing, LIU Hua, YANG Cuixia, SHEN Yunyue

2023, 38(9):  849-854.  DOI: 10.3969/j.issn.1673-8640.2023.09.007

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Objective To investigate the clinical value of serum inflammatory factors with prostate-specific antigen（PSA） and free prostate-specific antigen（f-PSA） in the auxiliary diagnosis of prostate cancer. Methods From July 2020 to November 2022，49 prostate cancer patients（prostate cancer group），50 prostate hyperplasia patients（benign prostate hyperplasia group） and 33 healthy subjects（healthy control group） were enrolled from Shanghai Jiao Tong University School of Medicine Affiliated Sixth People's Hospital. The serum levels of tumor necrosis factor-alpha（TNF-α），interleukin（IL）-1β，interleukin-2 rceptor（IL-2R），IL-8，IL-10，PSA and f-PSA were determined，and the clinical data of prostate cancer patients were collected. The efficacy of single and combined determinations of various indicators in the diagnosis of prostate cancer was evaluated by receiver operating characteristic（ROC） curve. Results The serum levels of TNF-α and IL-8 in prostate cancer group were higher than those in healthy control group and benign prostate hyperplasia group（P<0.05）. The serum levels of TNF-α in benign prostate hyperplasia group were higher than those in healthy control group（P<0.05）. There was no statistical significance in serum levels of IL-1β，IL-2R and IL-10 among the 3 groups（P>0.05）. The serum levels of TNF-α and IL-8 in prostate cancer patients with different ages，Gleason scores，TNM stages，the International Society of Urological Pathology（ISUP） grades，hypertension history，diabetes mellitus history and nerve invasion had no statistical significance（P>0.05）. Using the healthy control group and the benign prostate hyperplasia group as control，ROC curve analysis results showed that the areas under curves（AUC） of TNF-α，IL-8，PSA and f-PSA single determinations for diagnosing prostate cancer were 0.704，0.642，0.892 and 0.811，respectively，and the AUC for the combined determination of TNF-α，IL-8，PSA，f-PSA and f-PSA/PSA ratio was 0.946，which was higher than the other AUC（P<0.05）. Conclusions The combined determination of serum TNF- α，IL-8，PSA，f-PSA and f-PSA/PSA ratio can improve the diagnostic efficiency of prostate cancer.

Number and activity of peripheral blood eosinophils in obese people

CHENG Xu, YANG Cunqing, PANG Bo, GU Chun, HOU Xueyun, FEI Jiaxin, WU Min, LI Jun, LIU Guijian

2023, 38(9):  855-859.  DOI: 10.3969/j.issn.1673-8640.2023.09.008

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Objective To analyze the number and activity of peripheral blood eosinophils in obese people. Methods A total of 41 subjects >40 years old at Guang' anmen Hospital from June to September 2021 were enrolled. According to body mass index（BMI），the subjects were classified into obese group（20 cases，BMI≥30 kg/m²） and control group（21 cases，18.5 kg/m²<BMI<28 kg/m²）. The general information were collected. The expressions of CD62L and CD63 in peripheral blood eosinophils were determined by flow cytometry. The level of serum eosinophil cationic protein（ECP） was determined by enzyme-linked immunosorbent assay（ELISA）. The number and percentage of peripheral blood eosinophils and the expressions of CD62L，CD63 and ECP were compared between the 2 groups. Results The number and percentage of peripheral blood eosinophils and neutrophils and serum ECP levels in obese group were higher than those in control group（P>0.05）. CD62L was lower than that in control group（P<0.05），and CD63 was higher than that in control group（P<0.01）. CD62L was negatively correlated with BMI（r=-0.332，P=0.034）. Conclusions Obesity might be related to the increase of peripheral blood eosinophil activity.

Elevated red blood cell distribution width increasing the risk of heart failure：a cohort study based on the UK Biobank

LEI Jing, MAN Qiuhong, ZHAO Renjia, ZHANG Tiejun, JIANG Yanfeng, XU Kelin, SUO Chen, CHEN Xingdong

2023, 38(9):  860-864.  DOI: 10.3969/j.issn.1673-8640.2023.09.009

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Objective To understand the correlation between red blood cell distribution width（RDW） and heart failure（HF） in British population，and to provide new clues for the prevention and early diagnosis of HF. Methods The data from 466 420 UK Biobank participants，excluding those with the following prevalent diseases at baseline：HF，atrial fibrillation，myocardial infarction and angina pectoris，were collected. Patients with HF were defined using the International Classification of Diseases，10th edition，and diagnosis codes were I11.0，I130，I132 and I50. According to the quarters of RDW，they were classified into Q1（RDW<12.90%），Q2（RDW 12.90%-13.32%），Q3（RDW 13.33%-13.84%） and Q4（RDW≥13.85%） groups. Kaplan-Meier and Cox proportion hazard regression analysis were used to analyze the influence of RDW on the incidence of HF. Results During the follow-up period，a total of 5 362 HF patients were enrolled，with an incidence rate of 1.43‰，a male incidence rate of 2.05‰ and a female incidence rate of 0.93‰. Multivariate stepwise regression showed that sex，age，body mass index，blood pressure，Townsend deprivation index，smoking，healthy diet score and diabetes mellitus history were independent risk factors for HF. After adjusting the above variables，RDW was independently associated with the risk of HF [Q1 was used as control；Q2：hazard ratio（HR）=1.10，P=0.19；Q3：HR=1.24，P<0.05；Q4：HR=1.94，P<0.01]. Conclusions Elevated RDW is associated with an increased risk of developing HF and may be a potential early predictor of HF.

Role of Mycoplasma pneumoniae RNA determination in diagnosis and efficacy monitoring of pneumonia in children

WANG Huiying, TANG Yu, DONG Lili, WANG Jing

2023, 38(9):  865-869.  DOI: 10.3969/j.issn.1673-8640.2023.09.010

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Objective To analyze the role of Mycoplasma pneumoniae（MP） RNA determination in the diagnosis and efficacy monitoring of pneumonia in children. Methods Totally，167 children with pneumonia in Zhengzhou Children's Hospital from May 2019 to January 2021 were enrolled. Sputum samples were collected for MP-RNA，MP-DNA，MP-IgG antibody and MP-IgM antibody determinations. The specificities and sensitivities of MP-DNA，MP-RNA，MP-IgM antibody and MP-IgG antibody for diagnosing MP were evaluated. The positive rate of MP-RNA and MP-DNA and the incidence of clinical symptoms after 1，2，3 and 4 weeks of treatment were analyzed. The duration of clinical symptoms and serum C-reactive protein（CRP），interleukin-8（IL-8） and white blood cell（WBC） count of children with different MP-RNA turn-negative time were compared. Results MP culture proved that 95 of 167 children were infected with MP. The specificities for diagnosing MP pneumonia in children were MP-RNA>MP-DNA>MP-IgM antibody>MP-IgG antibody. The sensitivities were MP-RNA>MP-IgM antibody>MP-DNA>MP-IgG antibody. The positive rates of MP-RNA and MP-DNA were negatively correlated with the duration of treatment（r=-0.901，P=0.017；r=-0.601，P=0.031）. For 2，3 and 4 weeks of treatment，the positive rate of MP-RNA was lower than that of MP-DNA（P<0.000 1）. Compared with MP-DNA，the incidence of cough after MP-RNA turn-negative was higher（P<0.05）. After 1，2，3 and 4 weeks of treatment，the duration of fever，cough，wheezing and pulmonary rales in MP-RNA turn-negative children were successively prolonged（P<0.000 1），and serum CRP，IL-8 and WBC count were successively increased on admission（P<0.000 1）. Conclusions MP-RNA and MP-DNA have certain diagnostic value in children with MP pneumonia. In contrast，MP-DNA is more suitable for the diagnosis of children with MP，and MP-RNA is suitable for monitoring the efficacy of children with MP.

Bronchoscopy combined with sputum Mycobacterium tuberculosis ribonucleic acid and serum Mycobacterium tuberculosis specific antibody determinations for diagnosis of negative pulmonary tuberculosis

SUN Xinlin, YE Xingming, LI Qian, ZHAO Lijun

2023, 38(9):  870-873.  DOI: 10.3969/j.issn.1673-8640.2023.09.011

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Objective To investigate the efficiency of bronchoscopy combined with sputum Mycobacterium tuberculosis RNA（TB-RNA） and serum Mycobacterium tuberculosis specific antigen（TB-SA） determinations for the diagnosis of negative pulmonary tuberculosis. Methods A total of 104 patients with bacterial-negative pulmonary tuberculosis diagnosed by sputum Mycobacterium tuberculosis（MTB） culture in Huai'an Fourth People's Hospital from October 2018 to October 2019 were enrolled as observation group，and 80 patients with non-pulmonary tuberculosis during the same period were enrolled as control group. Bronchoscopy was performed，sputum samples were collected to determine sputum TB-RNA and serum TB-SA in sputum samples，and the diagnostic efficiency of each determination method for bacterial-negative pulmonary tuberculosis was evaluated. Results The positive rate of bronchoscopy in observation group（62.50%） was higher than that in control group（32.0%）（P<0.05）. The positive rate of sputum TB-RNA in observation group（78.85%） was higher than that in control group（22.50 %）（P<0.05）. The positive rate of serum TB-SA in observation group （68.27%）was higher than that in control group（20.00%）（P<0.05）. The sensitivity of the combined determination of the 3 methods was higher than those of bronchoscopy（P<0.001），TB-RNA determination（P=0.037） and TB-SA determination（P<0.001）. The accuracy of the combined determination of the 3 methods was higher than those of bronchoscopy（P<0.001），TB-RNA determination（P=0.028） and TB-SA determination（P<0.001）. There was no statistical significance in the specificity of the combined determination of the 3 methods（P>0.05）. Conclusions Bronchoscopy combined with sputum TB-RNA and serum TB-SA can effectively improve the sensitivity for the diagnosis of bacterial-negative pulmonary tuberculosis，which provides a determination method for the clinical diagnosis of bacterial-negative pulmonary tuberculosis，with high clinical application value.

Relationship between GeneXpert MTB/RIF assay determination load and Mycobacterium tuberculosis culture and phenotype of rifampicin resistance

MIAO Xingguo, YE Hui, SU Feifei

2023, 38(9):  874-877.  DOI: 10.3969/j.issn.1673-8640.2023.09.012

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Objective To analyze the relationship between GeneXpert MTB/RIF assay（Xpert）determination load and Mycobacterium tuberculosis（MTB） culture positive rate and rifampicin resistance phenotype. Methods From April 2016 to December 2019，the patients whose sputum Xpert determination results showed rifampicin resistance were enrolled from Wenzhou Central Hospital，and the differences between the positive rate of MTB and fasle positive rate for determining rifampicin resistance among patients with different Xpert determination loads were evaluated. Results Of the 183 patients，48，46，56 and 33 cases showed Xpert determination loads of high，medium，low and very low，respectively. Positive MTB culture was found in 80.87%（148/183） patients. The drug resistance rates of first-line anti-tuberculosis drugs were 79.05%（117/148）for isoniazid，81.08%（120/148） for rifampicin，27.03%（40/148） for ethambutol and 52.70%（78/148） for streptomycin. Totally，11 cases were fully sensitive to first-line anti-tuberculosis drugs. Among the 24 patients who were only resistant to 1 drug，the most common one was rifampicin（17 cases）. Totally，34 patients were resistant to 2 drugs，and rifampicin and isoniazid were the most common（25 cases）；53 patients were resistant to 3 drugs，among which rifampicin，isoniazid and streptomycin were the most common（40 cases）. The positive rates of MTB culture in patients with very low，low，medium and high Xpert determination loads were 48.48%，76.79%，93.48% and 95.83%，respectively. There was statistical significance in MTB culture's positive rates among patients with different Xpert determination loads（P<0.001）. The false positive rates of rifampicin phenotypes were 43.75%，23.26%，13.95% and 10.87% in patients with very low，low，medium and high Xpert determination loads，respectively. There was statistical significance in the false-positive rates of rifampicin phenotypes among patients with different Xpert determination loads（P=0.022）. Conclusions The determination of MTB with Xpert could reduce the missed diagnosis rate of culture-negative pulmonary tuberculosis. However，false-negative culture and false-positive rifampicin resistance are more likely to occur when the loads were low.

Correlation between small and dense low-density lipoprotein cholesterol and acute coronary syndrome with the occurrence of long-term adverse cardiovascular events

FU Yi, YANG Xuesong, JIN Ligang, ZHAI Hongli, JIN Xiaoling

2023, 38(9):  878-883.  DOI: 10.3969/j.issn.1673-8640.2023.09.013

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Objective To investigate the correlation between small and dense low-density lipoprotein cholesterol（sd-LDL-C） and acute coronary syndrome（ACS） with long-term adverse cardiovascular events in ACS patients. Methods Totally，149 ACS patients who received coronary artery stent implantation treatment were enrolled，which included 32 cases of acute myocardial infarction（AMI）（AMI group） and 117 cases of unstable angina pectoris（UAP）（UAP group）. Totally，50 healthy subjects were enrolled as control group. Total cholesterol（TC），triglyceride（TG），low-density lipoprotein cholesterol（LDL-C），high-density lipoprotein cholesterol（HDL-C），apolipoprotein（apo） A1，apo B and lipoprotein（a）[Lp（a）] were determined，and non-high-density lipoprotein cholesterol（non-HDL-C） was calculated. Logistic regression analysis was used to evaluate the risk factors for the occurrence of ACS. All the patients were followed up for 36 months. Kaplan-Meier survival curve was used to evaluate the incidence of long-term adverse cardiovascular events in ACS patients with different levels of sd-LDL-C. Cox regression analysis was used to evaluate the risk of major adverse cardiovascular events（MACE） and recurrent ACS in ACS patients with different levels of sd-LDL-C. Results The levels of serum TC，TG，LDL-C，non-HDL-C，apo B and sd-LDL-C in AMI and UAP groups were higher than those in control group（P<0.05），while the levels of serum HDL-C were lower than those in control group（P<0.05）. There was no statistical significance in apo A and Lp（a） among the 3 groups（P>0.05）. The results of Logistic regression analysis showed that sd-LDL-C was a risk factor for the occurrence of ACS [odds ratio（OR）=67.184，95% confidence interval（CI）5.284-854.213]. Based on the occurrence of adverse prognosis in ACS patients，receiver operating characteristic（ROC） curve was drawn，and all the ACS patients were classified into 2 groups，low sd-LDL-C group and high sd-LDL-C group，with the optimal cut-off value of 1.145 mmol/L. Kaplan-Meier survival curve showed that patients with high sd-LDL-C levels had a higher incidence of long-term adverse cardiovascular events compared to patients with low sd-LDL-C levels（P<0.05）. Cox regression analysis indicated that an increasing in sd-LDL-C had independent prognostic value for adverse cardiovascular events（total long-term adverse cardiovascular events：HR=4.376，95%CI 2.178-8.791；MACE：HR=3.848，95%CI 2.419-6.120；recurrent ACS：HR=4.067，95%CI 2.002-8.262）. Conclusions The sd-LDL-C has clinical value in early prevention and long-term prognosis evaluation of ACS.

Application of EWMA，HIQC and autoverification in primary medical institutions

MENG Xi, WEI Zhen, BAI Jianhua, HAO Xiaoke, ZENG Xianfei

2023, 38(9):  884-889.  DOI: 10.3969/j.issn.1673-8640.2023.09.014

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Objective To investigate the applicability of 3 quality management methods，exponentially weighted moving average（EWMA） of patient data，interlocalization of internal quality control（IQC） and automatic audit，in clinical laboratories of primary medical institutions. Methods The IQC data of 6 items including glucose，total cholesterol，triglyceride，total protein，albumin and urea were collected from 3 primary community health service centers under the supervision of Xi'an Area Medical Laboratory Center from June to November 2021. EWMA was used to analyze，and the application effect of EWMA was evaluated. Based on the standard deviation coefficient（SDI），coefficient of variation ratio（CVR） and other quality evaluation indexes，the consistency of results between clinical laboratories was compared. Automatic audit rules for 24 biochemical determination items were established，the coincidence rate and pass rate of manual audit and automatic audit were compared，and the differences in key indexes such as clinical laboratory sample turn-around time（TAT） before and after using automatic audit rules were compared as well. Results The trend of traditional IQC was consistent with the trend of EWMA quality control，EWMA can be offset earlier than traditional IQC identification and inspection system，but the uneven age distribution caused the false out-of-control alarm of EWMA. Among the 3 primary community health service centers，1 item（urea） SDI exceeded the allowable range and had serious negative deviation，which was consistent with the deviation trend of the results of external quality assessment（EQA） in the same period. The coincidence rate of automatic audit and manual audit during verification period was 100%，and the total pass rate of automatic audit during formal use was 95.33%. The median and 90th percentile of TAT were shortened by 24 and 136 min，respectively. Conclusions EWMA of patient data，interlocalization of IQC and automatic audit perform well in the quality management of primary medical institutions. They can sensitively identify the changes of clinical laboratory determination system，assist in out-of-control analysis，ensure the accuracy of clinical laboratory results and effectively shorten TAT.