Table of Content

30 August 2022, Volume 37 Issue 8

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Thoughts on whether adenovirus is the cause of the hepatitis of unknown etiology among children recently

LU Lijuan, XU Jin

2022, 37(8):  705-709.  DOI: 10.3969/j.issn.1673-8640.2022.08.001

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Recently,the hepatitis of unknown etiology among children has shown a concentrated trend in many countries and regions,which has aroused concern from the World Health Organization. Based on the reports of existing cases,numerous hypotheses have been proposed,among which the hypothesis that adenovirus may be the cause of the hepatitis of unknown etiology among children is debated widely. According to the occurrence of the hepatitis of unknown etiology among children,the pathogenic characteristics of adenovirus and literatures on adenovirus hepatitis,this review analyzes the pathogens that may cause the hepatitis of unknown etiology among children and puts forward suggestions on the work to be carried out to clarify the pathogens. All of those details may provide a direction for timely and accurate diagnosis of related cases in China.

Analysis of perinatal clinical features and genetic characteristics of a neonate with otopalatodigital syndrome type 1

ZHAO Xuliang, TIAN Ruixia, LI Xu, JIA Jian'an, YU Min, ZHU Fuxi

2022, 37(8):  710-714.  DOI: 10.3969/j.issn.1673-8640.2022.08.002

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Objective To investigate the perinatal clinical features and genetic characteristics of a neonate with otopalatodigital syndrome type 1（OPD1）,and to improve the clinical understanding of such rare diseases. Methods The clinical data of a neonate with OPD1 was collected. The genes of the neonate and his parents were determined by trio-whole exome sequencing（Trio-WES）. Variation sites were verified by Sanger sequencing. The biohazard of gene mutation was analyzed by relevant prediction software. The clinical features of the neonate with filamin A（FLNA） gene hot spot mutation were analyzed retrospectively. Results Prenatal ultrasound showed that the fetal had clinical manifestations such as micrognathia,nasal collapse,abnormal spinal alignment and the length of the second toes of both feet. After birth,the case had a special appearance of ocular hypertelorism,collapsed nasal bridge and wide nasal root,small mandible and backward retraction,low ear position and accessory ear. The results of gene test showed that the FLNA gene had heterozygous mutation（c.620C>T/p.Pro207Leu）,his mother carried the same variant,and his father has wild type. Literature reviewing showed that FLNA gene c.620C>T/p.Pro207Leu was the hot spot variant causing to OPD1. The high frequency phenotypes of the mutation was ocular hypertelorism,eyelid cleft,micrognathia,cleft palate,different types of finger（toe） deformity and hearing loss. Conclusions OPD1 is caused by FLNA gene hot spot variant c.620C>T/p.Pro207Leu. The combination of family datum analysis,prenatal ultrasound and family gene analysis is an effective method for the diagnosis and further typing of otopalatodigital spectrum disorder（OPDSD）.

Correlation of serum exosomes,placental exosomes and exosomal miR-210 with preeclampsia

DUAN Yuping, XIE Li, CAI Leiming, WU Jingjing, HONG Mao, LI Qian

2022, 37(8):  715-719.  DOI: 10.3969/j.issn.1673-8640.2022.08.003

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Objective To investigate the changes and roles of serum exosomes,placental exosomes [placental alkaline phosphatase（PLAP）] and exosomal microRNA-210（miR-210） in pregnant women during preeclampsia（PE） pregnancy. Methods Totally,30 patients with PE and 30 healthy pregnant women were enrolled. All serum samples were collected in the first trimester,the second trimester and the third trimester of pregnancy. The amount of serum exosomes,the level of PLAP and the expression of exosomal miR-210 were determined. The correlation between PLAP and miR-210 was analyzed using Pearson analysis. Results The amount of serum exosomes,the level of PLAP and miR-210 were increased with the progress of pregnancy. Compared with healthy pregnant women,the amount of serum exosomes,the level of PLAP and miR-210 in PE group were higher in different trimesters of pregnancy（P<0.05）. The correlation between the level of PLAP and exosomal miR-210 was positive（r=0.668,P<0.01）. Conclusions The amount of serum exosomes,the level of PLAP and exosomal miR-210 in PE group change from the first trimester of pregnancy,and they play roles in the early prediction and diagnosis of PE.

Construction and clinical application of colon cancer prognostic risk model of ferroptosis-related lncRNA

WANG Yuqing, ZHANG Yue, XU Wen, CUI Zhongqi

2022, 37(8):  720-728.  DOI: 10.3969/j.issn.1673-8640.2022.08.004

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Objective Based on the Cancer Genome Atlas（TCGA） database,ferroptosis-related long non-coding RNA（lncRNA） were screened to establish a colon cancer prognostic risk model and to investigate its clinical application role. Methods A total of 48 patients who underwent radical resection of colon cancer were enrolled,and the intraoperatively resected cancer tissues and corresponding paracancerous tissues（2-3 cm from the edge of the cancer tissue）were collected. Transcriptome data of colon cancer（428 cases of colon cancer tissues and 41 cases of normal colon tissues）and clinical data of corresponding patients were collected from the TCGA database. Gene Ontology（GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG）pathway analysis were performed for ferroptosis-related genes differentially expressed in colon cancer tissues and normal colon tissues. Screening for ferroptosis-related lncRNA with colon cancer prognosis,consensus clustering analysis was used to classify colon cancer patients and compare overall survival. A prognostic risk model was established by LASSO-Cox regression analysis. A nomogram for predicting overall survival in patients with colon cancer was developed. The biological function of the key lncRNA ITGB1-DT in colon cancer was analyzed by cytological experiments. Results Totally,72 differentially expressed ferroptosis-related genes were screened from the TCGA database,of which 47 cases were up-regulated,and 25 cases were down-regulated. GO enrichment analysis and KEGG pathway analysis showed that these genes were widely involved in biological processes,such as iron metabolism and fatty acid oxidation. A total of 20 ferroptosis-related lncRNA with difference between colon cancer tissues and normal colon tissues were screened. There was statistical significance in overall survival between the 2 colon cancer subgroups classified according to the results of consensus clustering analysis（P<0.001）. According to the results of LASSO-Cox regression analysis,10 lncRNA were screened,and a prognostic risk model was constructed. A total of 428 patients with colon cancer were randomly classified into training group and validation group according to the ratio of 7∶3. In the training group,validation group and overall colon cancer patients,the overall survival rate of high-risk group was lower than that of low-risk group（P<0.001）. The relative expression of ITGB1-DT in colon cancer tissues was higher than that in adjacent tissues（P<0.001）. The results of cytological experiments showed that ITGB1-DT could promote the proliferation of colon cancer cells and inhibit its ferroptosis. Conclusions A colon cancer prognostic risk model based on ferroptosis-related lncRNA has been constructed successfully,and a nomogram that could be used to judge the overall survival rate of colon cancer has been established. ITGB1-DT may promote the development of colon cancer.

Effects of SARS-CoV-2 vaccination on antibody production and immune function

TAN Meiyu, SHU Jie, XUAN Binbin, ZHOU Lida, LI Hong, HOU Shangwei, SHENG Huiming

2022, 37(8):  729-734.  DOI: 10.3969/j.issn.1673-8640.2022.08.005

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Objective To investigate the changes of antibody production and immune function in healthy subjects after receiving novel coronavirus-inactivated vaccine（Vero cell）. Methods Totally,121 volunteers were classified into two dose and three dose groups. Severe acute respiratory syndrome coronavirus 2（SARS-CoV-2） total antibodies（IgG+IgM+IgA）,SARS-CoV-2 IgG,SARS-CoV-2 receptor binding domain（RBD） specific neutralizing antibody,lymphocyte subsets and 12 cytokine levels were determined before the first dose,at 7 d after the second dose,before the third dose（half a year after the second dose） and at 7 d after the third dose. Results After the volunteers received the vaccine in the two dose group,the positive rates of SARS-CoV-2 total antibody and SARS-CoV-2 IgG were all 93.44%,and the positive rate of neutralizing antibody of SARS-CoV-2 was 81.96%. For the three dose group,the positive rate of SARS-CoV-2 total antibody was 100.00%,and the positive rate of SARS-CoV-2 IgG was 93.33%. The total antibody level of SARS-CoV-2 in the three dose group at 7 d after the third dose was higher than that in the two dose group at 7 d after the second dose（P<0.001）,and the neutralizing antibody level was also statistically significant（P<0.01）. The percentage of T follicular helper cell（Tfh） in the two dose group at 7 d after the second dose（11.40%） was higher than that before the first dose（3.92%）（P<0.01）. There was no statistical significance in lymphocyte subsets and 12 cytokine levels before the first dose and at 7 d after the second dose in the two dose group,and there was no statistical significance in the three dose group（P>0.05）. Conclusions Neutralizing and reactive antibodies can be produced after vaccination with novel coronavirus-inactivated vaccine（Vero cell）. Vaccination does not affect the immune function of the humans.

Correlation of lung cancer clinicopathological characteristics with FR⁺-CTC,ANXA2 and ProGRP

LIU Xingqiang, NING Lifen, LI Lin, CHEN Zhongcheng

2022, 37(8):  735-740.  DOI: 10.3969/j.issn.1673-8640.2022.08.006

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Objective To investigate the clinical roles of folate receptor positive circulating tumor cells（FR⁺-CTC）,annexin A2（ANXA2） and gastrin-releasing peptide precursor（ProGRP） in lung cancer. Methods A total of 116 lung cancer patients（lung cancer group） and 100 benign lung disease patients（control group） were enrolled. The levels of FR⁺-CTC,ANXA2,ProGRP,cytokeratin 19 fragment（CYFRA21-1）,neuron specific enolase（NSE） and squamous cell carcinoma antigen（SCC-Ag） were determined. Receiver operating characteristic（ROC） curve was used to evaluate the efficacy of each index in the diagnosis of lung cancer. Spearman correlation analysis was used to evaluate the correlation among the indicators. Multivariate Logistic regression analysis was used to evaluate the risk factors of lung cancer. Results The levels of FR⁺-CTC,ANXA2 and ProGRP in lung cancer group were higher than those in control group（P<0.05）. ROC curve results showed that the areas under curves（AUC） of FR⁺-CTC,ANXA2 and ProGRP single and combined determinations were 0.885,0.810,0.696 and 0.935,respectively. Spearman correlation analysis showed that FR⁺-CTC,ANXA2 and ProGRP were positively correlated with CYFRA21-1,NSE and SCC-Ag（P<0.05）. FR⁺-CTC,ANXA2 and ProGRP were increased in patients with smoking history,TNM stage Ⅲ-Ⅳ,tumor diameter ≥5 cm,low differentiation,lymph node metastasis and distant metastasis（P<0.05）. The positivities of FR⁺-CTC,ANXA2 and ProGRP were risk factors for lung cancer [odds ratios（OR） were 1.385,1.774 and 1.823,95% confidence intervals（CI） were 1.187-4.974,1.601-5.075 and 1.677-6.005,respectively]. Conclusions The levels of FR⁺-CTC,ANXA2 and ProGRP are related to the clinicopathological characteristics of lung cancer,and they may be used for the diagnosis and evaluation of lung cancer.

Analysis on outcome of patients with indeterminate western-blot results of HIV-1 antibody

SUN Qi, QU Xinyi, XU Li

2022, 37(8):  741-744.  DOI: 10.3969/j.issn.1673-8640.2022.08.007

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Objective To investigate the characteristics and outcomes of patients with indeterminate western-blot results of human immunodeficiency virus（HIV）-1 antibody. Methods Totally,75 patients with indeterminate western-blot results of HIV-1 antibody were enrolled. The general data,screening test results [enzyme-linked immunosorbent assay（ELISA）and colloid se method],western-blot results and the outcomes of follow-up were collected. Results In the 75 cases,53 cases were followed up,and the follow-up time was 10-721 d. The difference of positive conversion rate between males and females had statistical significance（P<0.05）. Among the 53 cases,the positive consistency rate of the 2 screening tests was 100%（45 cases）. Using receiver operating characteristic（ROC） curve analysis,the optimal cut-off value of ELISA S/CO value was 4.995,the area under curve（AUC） was 0.969,the sensitivity was 84.8%,and the specificity was 100.0%. With the increase of ELISA S/CO values,the western-blot bands turned into positive correspondingly,and the occurrence rates of env gp41,gag p66 and p51 were also increased correspondingly. Conclusions The collaborative analysis of colloid se method,ELISA S/CO value and western-blot band will be helpful for the determination of indeterminate HIV-1 antibody.

Screening and bioinformatics analysis of differential genes in children with acute lymphoblastic leukemia based on GEO database

YU Lisha, ZHANG Yingying

2022, 37(8):  745-750.  DOI: 10.3969/j.issn.1673-8640.2022.08.008

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Objective To analyze the differential expression genes between children with acute lymphoblastic leukemia（ALL） and healthy children by bioinformatics,and to find new molecular markers for the auxiliary diagnosis of ALL in children. Methods The gene chips of newly diagnosed children with ALL and healthy children was downloaded from the Gene Expression Omnibus（GEO）,and the differential expression genes were screened by R language. The Database for Annotation,Visualization and Integrated Discovery（DIVID） was employed for Gene Ontology（GO） and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis. The protein interaction network of differential expression genes was constructed on STRING database,the results were visualized by Cytoscape software,and the hub genes were screened by CytoHubba plug-in. Results A total of 2 platforms' gene chips of newly diagnosed ALL children and healthy children were included,and 245 common differential expression genes were selected,which included 88 up-regulated genes and 157 down-regulated genes. Enrichment analysis showed that the common differential expression genes were mainly concentrated in the immune response（biological process）,integrating in the extracellular space（cell composition）,participating protein binding（molecular function）,and enriching hematopoietic cell lineage and cell cycle signal pathways（KEGG pathway analysis）. Totally,10 hub genes were screened out,namely CDK1,TOP2A,TYMS,MCM2,MCM4,TTK,CCNB2,BUB1B,KIF4A and MAD2L1. Conclusions The bioinformatics analysis of children with ALL reveals that 10 hub genes may be new markers for assisting the diagnosis of children with ALL.

Methodology evaluation of screening techniques for CRE colonized in intestinal tract

XU Qi, PAN Fen, SUN Yan, SHI Yingying, YU Fangyuan, ZHANG Hong

2022, 37(8):  761-765.  DOI: 10.3969/j.issn.1673-8640.2022.08.012

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Objective To evaluate the performance of screening methods by home-made plate containing meropenem 1 μg/mL（drug screening method） and MacConkey agar with 10 μg meropenem（disk screening method） in the screening of carbapenem-resistant Enterobacteriaceae（CRE） colonized in intestinal tract. Methods A total of 306 stool samples for CRE screening were collected from hospitalized children between July 2019 and March 2020,and drug and disk screening methods were used to determine CRE isolates colonized in stool samples. The results of polymerase chain reaction（PCR） amplified carbapenemase genes（PCR screening method） were taken as a standard to evaluate the performance of the 2 screening methods mentioned above. Results Among the 306 stool samples,the positive rate of PCR,drug and disk screening methods were 12.1%（37/306）,17.3%（53/306）and 15.4%（47/306）,respectively. The PCR screening method was used as a standard,the sensitivity,specificity,positive predictive value,negative predictive value and consistency of drug screening method were 97.3%,93.7%,67.9%,99.6% and 94.1%,respectively,and the Kappa value was 0.77（P<0.05） with good consistency. The sensitivity,specificity,positive predictive value,negative predictive value and consistency of disk screening method were 91.9%,95.2%,72.3%,98.8% and 94.8%,and the Kappa value was 0.78（P<0.05） with good consistency. There were 6 samples as positive both by drug and disk screening methods,but the carbapenemase genes were negative by PCR screening method. Considering CRE isolates might have other drug resistance mechanisms,it should consider those samples as positive for CRE screening. Conclusions The drug and disk screening methods can be applied to screen and determine the CRE colonized in intestinal tract. Clinical laboratories can select the corresponding methods according to the specific situation.

Comparison of testing efficiency of simulated emergency biochemical samples among different instruments GE

Danhong , FANG Huiling, LIN Feiran, ZHU Yuqing

2022, 37(8):  766-771.  DOI: 10.3969/j.issn.1673-8640.2022.08.013

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Objective To compare the testing efficiency of simulated emergency biochemical samples among different instruments. Methods Single analyte and full-set analytes for single sample and a batch of samples（40 samples） were tested with 3 different instruments（A,B,C）,respectively. The testing times of different instruments were recorded. Totally,20 samples were tested in different sequences（sequence 1 and sequence 2）,and the testing time were recorded. Totally,5 instrument combinations（Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅴ） were used to test biochemical analytes and cardiac markers for 10 samples. The combinations were determined according to various processes,and the testing times were recorded. Both split sample and unsplit sample were determined on the combinations. The samples were determined in the process of biochemical analytes first or cardiac markers first. A total of 40 samples,containing 5 hemolytic,5 icteric and 5 lipemic samples,were collected,and the 40 samples were also inspected by 4 technicians by visual. The testing time and results were recorded,and the coincidence rates of visual results were calculated. Daily maintenance for the 3 instruments was performed,and the testing time was recorded. Results It took 6,16 and 12 min to test glucose for single sample for instrument A,B and C,respectively,and 8,18 and 14 min to test full-set analytes for the above instruments,respectively. It took 12,19 and 15 min to test glucose for a batch of samples for instrument A,B and C,respectively,and 103,112 and 72 min to test full-set analytes. It took the same time to test the same analytes for 10 samples in different sequences for instrument A and C,while it took 1 more min in sequence 2 than in sequence 1 on instrument B. When 10 samples were tested for biochemical analytes and cardiac markers,it took combination Ⅰ 34 min. It took combination Ⅱ,Ⅲ,Ⅳ and Ⅴ 67,60,92 and 80 min,respectively,when testing biochemical analytes firstly. It took 69,53,72 and 71 min respectively when testing cardiac markers firstly. It took 58,48,69 and 69 min when the samples were split and start testing simultaneously on both instruments. It took the same time for the 3 instruments to test all the biochemical analytes for 40 samples. The average time for visual inspection of 40 serum samples was 122 s. The mean coincidence rates of visual inspection for hemolytic,icteric and lipemic samples were 100%,48% and 76%,respectively. It took 13,21 and 35 min to complete daily maintenance for instrument A,B and C. Conclusions The testing time is influenced by the number of samples,testing process and so on. Serum index testing needs less time and labor than visual inspection. It reflects the status of samples more objectively and helps controlling the error of pre-analysis. Laboratories can improve the testing efficiency of instrument by means of rationally configuring instruments and optimizing testing process,which may help to shorten turn-around time（TAT）.

Mechanism of glioblastoma multiforme proliferation and invasion analyzed by whole-transcriptome sequencing

WANG Shichao, LIU Bin, ZHAO Xing, REN Xiaomin, PAN Yuanyuan, LI Xia, XU Shuying

2022, 37(8):  772-781.  DOI: 10.3969/j.issn.1673-8640.2022.08.014

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Objective To analyze the transcriptome sequencing and biological information data of glioblastoma multiforme（GBM） tissues and adjacent tissues,to dig out the key genes that cause GBM and identify their functions,and to investigate the molecular regulatory mechanism of malignant proliferation,invasion and migration of GBM. Methods Totally,4 patients with GBM were enrolled,and whole-transcriptome sequencing for tumor tissues and adjacent tissues was performed. The mRNA,microRNA（miRNA）,long non-coding RNA（lncRNA） and circular RNA（circRNA） with different expressions were screened. Bio-informatics [Gene Ontology（GO） enrichment analysis,Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway analysis,transcript enrichment analysis and so on] was performed. The mRNA-miRNA-lncRNA/circRNA associated network was established,and glioma development,migration and invasion related to target molecules were found. Results There were 3 088 different mRNA between tumor tissues and adjacent tissues,of which 1 375 cases were up-regulated,and 1 713 cases were down-regulated. There were 106 different lncRNA,of which 52 cases were up-regulated,and 54 cases were down-regulated. There were 82 different circRNA,of which 32 cases were up-regulated,and 50 cases were down-regulated. There were 15 different miRNA,of which 8 cases were up-regulated,and 7 cases were down-regulated. Differentially expressed mRNA transcripts were enriched to obtain 19 significant pathways. These pathways included Shh（Sonic hedgehog） signaling pathway,glioma signaling pathway,pancreatic cancer signaling pathway,axon guidance pathway,tumor necrosis factor（TNF） signaling pathway and other related pathways. The multivariate association analysis of whole-transcriptome sequencing found that BOC was located at the key node of its regulatory pathway（has-miR-4763-5p,has-miR-6848-5p and has-miR-6860 miRNA）. The mRNA expression level of BOC in tumor tissues was higher than that in adjacent tissues. Conclusions BOC may affect the expression of downstream target genes by the regulation of related miRNA,circRNA,lncRNA and other non-coding RNA,leading to the proliferation and invasion of GBM.

Research progress of body fluid piRNA as a potential biomarker of disease

WU Yanqian, YU Chong, SHEN Lu, ZHENG Haoran, HONG Yeting

2022, 37(8):  782-786.  DOI: 10.3969/j.issn.1673-8640.2022.08.015

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PIWI-interacting RNA（piRNA）is a new class of small non-coding RNA that can silence transposon and regulate mRNA translation. Previous studies have shown that piRNA is mainly involved in germ cell development and spermatogenesis,and it is closely related to tumorigenesis and other diseases. The differential expression levels of piRNA in body fluid under specific disease states suggest that piRNA have the potential to be a new biomarker. The review focuses on the progress of research on body fluid piRNA as a potential biomarker for various diseases.

Research progress of carbon quantum dots and graphene quantum dots in sensing field

LI Kun, ZHANG Guojun

2022, 37(8):  787-792.  DOI: 10.3969/j.issn.1673-8640.2022.08.016

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Carbon quantum dots and graphene quantum dots,which are 0 dimensional nanomaterials,have been widely used in sensing and determination due to their good conductivity,adjustable fluorescence,low toxicity,small size,ease of modification and so on. This review briefly introduces the basic properties of carbon quantum dots and graphene quantum dots,and mainly introduces the application in the field of sensing. The application prospect of carbon quantum dots and graphene quantum dots in laboratory medicine is briefly discussed.

Clinical characteristics and molecular epidemiology of Streptococcus agalactiae infection in non-pregnant adults

LIU Jingxian, CHEN Xingyue, ZHAO Jing, LIU Ying

2022, 37(8):  793-797.  DOI: 10.3969/j.issn.1673-8640.2022.08.017

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Streptococcus agalactiae has known as the leading cause of invasive disease in neonates and pregnant women. However,the morbidity of Streptococcus agalactiae infection in non-pregnant adults is increasing rapidly. The clinical characteristics of Streptococcus agalactiae infection and the molecular epidemiology of Streptococcus agalactiae isolated from non-pregnant adults have rare understood. The review provides a reference for the diagnosis,treatment prevention and control of this disease.