Table of Content

30 March 2022, Volume 37 Issue 3

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Evaluation of nasopharyngeal swab bacterial culture in pathogenic diagnosis in children with bacterial respiratory tract infection

XIE Yongping, HUA Chunzhen, WEI Linlin, WANG Hongjiao, WANG Gaoliang, LI Jianping

2022, 37(3):  201-204.  DOI: 10.3969/j.issn.1673-8640.2022.03.001

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Objective To evaluate the value of nasopharyngeal swab bacterial culture in pathogenic diagnosis in children with respiratory tract infection. Methods Nasopharyngeal swab and oral aspirate samples were collected from 403 hospitalized children diagnosed as pertussis-like syndrome or pneumonia. The samples were inoculated in partitions on medium discs with semi-quantitative method. Bacteria were identified,and the isolates growing including and more than "+++" were definited as clinical isolates. The positive rates of culture between nasopharyngeal swab and sputum samples were compared and linked with clinical manifestation and treatment outcomes. Results In the 403 cases,the positive rate of nasopharyngeal swab culture was 31.5%（127/403）,and the top 4 bacteria were Bordetella pertussis（13.4%）,Haemophilus influenzae（7.9%）,Moraxella catarrhalis（4.0%） and Streptococcus pneumoniae（3.5%）. The positive rate of oral aspirate culture was 12.4%（50/403）,and the positive rates of the above 4 bacteria were 0.7%,4.5%,1.0% and 1.2%,respectively. The positive rates of nasopharyngeal swab culture was higher than that of oral aspirate culture（P<0.05）. The same bacteria were isolated from both nasopharyngeal swab and sputum samples in 31 of 50 cases（62.0%）. After treating with sensitive antibiotics,the results of nasopharyngeal swab culture swithed to negative in 98.4%（125/127） of all the patients. Conclusions Nasopharyngeal swab culture improves the determination of pathgens in respiratory tract in children,which plays a role in pathogenic diagnosis in children with respiratory tract infection.

Drug susceptibility of multidrug-resistant ESBL-producing Escherichia coli to antimicrobial agents restored by different concentrations of EDTA

WEN Qinghui, LI Fengying, LI Yanchang, HUANG Jinyin, WANG Fengping

2022, 37(3):  205-208.  DOI: 10.3969/j.issn.1673-8640.2022.03.002

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Objective To investigate the effect of different concentrations of ethylenediaminetetraacetic acid（EDTA） on the recovery of drug susceptibility of multidrug-resistant extended-spectrum beta-lactamase（ESBL）-producing Escherichia coli. Methods Totally,240 isolates of ESBL-producing Escherichia coli were collected. The minimum inhibitory concentrations（MIC） before and after adding EDTA with low（0.5 mg/mL）,standard（1.0 mg/mL） and high（2.0 mg/mL） concentrations were determined. Results Among the isolates of ESBL-producing Escherichia coli,the difference between the MIC of pure antibiotics and the MIC of antibiotics with EDTA was obvious. The changes of MIC of cefoperazone,ofloxacin and cefepime were the most obvious,followed by amikacin,tobramycin and gentamicin. The change of MIC of aztreonam was small,and the MIC of cefotaxime did not change. In the 3 groups of combination of EDTA and antibiotics at different concentrations,the changes in the MIC of antibiotics were more significant when the antibiotics were combined with the standard concentration and the high concentration of EDTA than when the antibiotics were combined with the low concentration of EDTA,but there was no statistical significance between the standard concentration and the high concentration of EDTA. The MIC of cefotaxime did not change in the 3 groups. Conclusions EDTA of different concentrations has synergistic antibacterial effect on most determined isolates,and it can improve the drug susceptibility of antimicrobial agents,which has important clinical significance.

Influence of delayed entry on fastidious bacterium culture in 2 blood culture systems

CHU Minjun, LI Xiaojuan, HUANG Hailin, LI Jue, MA Mingbiao, DU Tingyi, WANG Haiping

2022, 37(3):  209-212.  DOI: 10.3969/j.issn.1673-8640.2022.03.003

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Objective To compare the effects of delayed entry on the positive detection rate and time of fastidious bacterium culture in 2 blood culture systems. Methods Simulated blood samples of Streptococcus pneumoniae and Haemophilus influenzae standard isolates with different concentrations were inoculated into BactecPeds Plus and BACT//ALERT PF culture flask at room temperature for 0,8,16 and 24 h,and then they were placed into the above 2 blood culture systems（BACTEC FX and BACT/ALERT 3D）. The culture results and positive times were recorded for statistical analysis. Results After being placed for different periods,positive results were determined in the all simulated blood samples,and the positive time of BACTEC FX was shorter than that of BACT/ALERT 3D（P<0.001）. After the delayed entry for 8,16 and 24 h,the positive time of Streptococcus pneumoniae was longer than that of the sample placed immediately（0 h）（P<0.05）. There were interactions between the delayed entry time,and the concentration,the blood culture systems（P<0.05）. There was no statistical significance in the positive time of Haemophilus influenzae between 0 h and 8 h of delayed entry（P>0.05）,and the delayed entry time,and the concentration,the blood culture systems had no interaction（P>0.05）. Conclusions The positive time of fastidous bacterium increases with the extension of the delayed entry time,which was influenced by multiple factors,such as strain,concentration and blood culture system.

Influence of length of supine position on the diagnosis of primary aldosteronism by ARR

JIN Yinxin, LI Zaizhao, YUAN Lingqing, FEI Dongxue, WANG Hui, JIANG Kunfang

2022, 37(3):  213-216.  DOI: 10.3969/j.issn.1673-8640.2022.03.004

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Objective To investigate the roles of plasma aldosterone level,renin activity and aldosterone/renin ratio（ARR） in different-time supine and standing positions on the diagnosis of primary aldosteronism（PA）. Methods A total of 103 patients with suspected PA were enrolled,which included 44 cases of PA group and 59 cases of non-PA group. Blood samples were taken for standing position 2 h,supine position 4 h and supine position 2 h,and plasma aldosterone level and renin activity were determined,and ARR was calculated. Receiver operating characteristic（ROC）curve was used to analyze the diagnostic value of ARR for PA. Results Between PA group and non-PA group,aldosterone levels,renin activities and ARR for standing position 2 h,supine position 4 h and supine position 2 h had statistical significance（P<0.01）. In PA group,aldosterone level,renin activity and ARR for standing position 2 h had statistical significance compared with those for supine position 4 h and supine position 2 h（P<0.05）. Within PA and non-PA groups,aldosterone level,renin activity and ARR for standing position 2 h,supine position 4 h and supine position 2 h had no statistical significance（P>0.05）. Aldosterone level,renin activity and ARR between supine position 4 h and supine position 2 h had no statistical significance（P>0.05）. ROC curve analysis results showed that the areas under curves（AUC） of ARR for standing position 2 h,supine position 4 h and supine position 2 h were 0.821,0.779 and 0.787,respectively. Conclusions ARR for standing position 2 h,supine position 4 h and supine position 2 h has certain accuracy in the diagnosis of PA. The test execution rate can be improved by supine position 2 h if the patients cannot lie for a long time.

Roles of total IgE,sIgE and 25（OH）D in the evaluation of infant atopic dermatitis

ZHANG Zhibin, YANG Wanyong, RUAN Fuwang, WEN Qinghui

2022, 37(3):  217-220.  DOI: 10.3969/j.issn.1673-8640.2022.03.005

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Objective To investigate the roles of total immunoglobulin E（IgE）,specific immunoglobulin E（sIgE） and 25-hydroxyvitamin D [25（OH）D] in the evaluation of infant atopic dermatitis（AD）. Methods A total of 120 infants with AD were enrolled and classified into mild group（39 cases）,moderate group（64 cases） and severe group（17 cases） according to the severity of the disease and into non-complication group（51 cases） and complication group（69 cases） according to the complications. Totally,120 healthy infants were enrolled as control group. Serum total IgE,sIgE and 25（OH）D levels were determined. Risk factors for severe AD were assessed by Logistic regression analysis. Results The level of serum 25（OH）D in AD group was lower than that in control group（P=0.000）,and the levels of total IgE and sIgE were higher than those in control group（P=0.000）. The levels of serum 25（OH）D in mild,moderate and severe groups decreased in turn（P=0.000）,and the levels of serum total IgE and sIgE increased in turn（P=0.000）. The level of serum 25（OH）D in complication group was lower than that in non-complication group（P=0.000）,and the levels of serum total IgE and sIgE were higher than those in non-complication group（P=0.000）. Logistic regression analysis showed that total IgE,sIgE and 25（OH）D were risk factors for severe AD in infants [odds ratios（OR） were 35.023,15.502,4.993,95% confidence intervals（CI） were 17.431-70.370,6.900-34.830,2.456-10.150,respectively]. Conclusions Serum 25（OH）D,total IgE and sIgE are correlated with the severity of AD in infants,or they can be used as indicators to judge the severity of AD in infants.

Changes and clinical significance of IP-10 for antiviral therapy in AIDS patients

FAN Weiguang, SU Miaomiao, MENG Juan, SHI Penghui, ZHANG Zhen

2022, 37(3):  221-225.  DOI: 10.3969/j.issn.1673-8640.2022.03.006

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Objective To investigate the changes and clinical significance of interferon-inducible protein-10（IP-10） for antiviral therapy in acquired immunodeficiency syndrome（AIDS） patients. Methods Totally,200 AIDS patients（AIDS group） receiving highly active antiroviral therapy（HAART） were enrolled and classified into HIV-1 inhibition group and HIV-1 non-inhibition group according to curative effect. The percentage of CD4⁺ T cells,the percentage of CD8⁺ T cells,the virus load of human immunodeficiency virus（HIV）-1 and the level of IP-10 were determined before treatment and at 2 weeks,1 month,2 months,3 months,6 months,12 months and 18 months after treatment. A total of 100 healthy subjects were enrolled as healthy control group. Spearman correlation analysis was used to evaluate the correlation between IP-10 and the other indicators. Results The level of HIV-1 RNA in AIDS group before treatment was（4.62±0.73） lg copies/mL. The percentage of CD4⁺ T cells in AIDS group was lower than that in healthy control group（P<0.01）. The level of IP-10 and the percentage of CD8⁺ T cells were higher than those in healthy control group（P<0.01）. Compared with those before treatment,with the extension of treatment time,the percentage of CD4⁺ T cells increased gradually（P<0.01）,the percentage of CD8⁺ T cells and the levels of IP-10 and HIV-1 RNA decreased gradually（P<0.01）,and the proportion of patients with inhibited HIV-1 replication increased gradually（P<0.01）. At 3,6,12 and 18 months of treatment,the percentage of CD4⁺ T cells in HIV-1 inhibition group was higher than that in HIV-1 non-inhibition group（P<0.01）,and the percentage of CD8⁺ T cells and IP-10 level were lower than those in HIV-1 non-inhibition group（P<0.01）. Spearman correlation analysis showed that the level of IP-10 in AIDS patients was negatively correlated with the percentage of CD4⁺ T cells（r=-0.763,P<0.05）,and it was positively correlated with the percentage of CD8⁺ T cells and the level of HIV-1 RNA（r=0.801,P<0.05;r=0.823,P<0.05）. Conclusions The level of IP-10 in AIDS patients is related to the levels of CD4⁺ T cells,CD8⁺ T cells and HIV-1 RNA. It is an indicator of potential AIDS and efficacy evaluation.

Roles of serum sICAM-1 and hs-CRP levels in monitoring acute viral lower respiratory tract infection in children

LI Dongxiu, HUANG Jian

2022, 37(3):  226-229.  DOI: 10.3969/j.issn.1673-8640.2022.03.007

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Objective To investigate the roles of serum soluble intercellular adhesion molecule-1（sICAM-1） and high-sensitivity C-reactive protein（hs-CRP） levels in monitoring acute viral lower respiratory tract infection in children. Methods Totally,136 children with acute viral lower respiratory tract infection were enrolled and classified into mild-moderate group（82 cases） and severe group（54 cases） according to severity. According to specific IgG antibody level,they were classified into acute phase and recovery phase groups. According to acute physiology and chronic health evaluation Ⅱ（APACHEⅡ）score,children were classified into low-risk group（39 cases）,intermediate-risk group（57 cases） and high-risk group（40 cases）. Serum sICAM-1,hs-CRP levels and oxygenation index were determined. Totally,50 healthy children were enrolled as healthy control group. Pearson correlation analysis was used to evaluate the correlation between various indicators. Results In acute phase,serum sICAM-1 and hs-CRP levels in severe group and mild-moderate group were higher than those in healthy control group（P<0.05）,and serum sICAM-1 and hs-CRP levels in severe group were higher than those in mild-moderate group（P<0.05）. The levels of serum sICAM-1 and hs-CRP in low-risk group,intermediate-risk group and high-risk group increased in turn（P<0.05）. The levels of serum sICAM-1 and hs-CRP in acute and recovery phases of severe group were higher than those of mild-moderate group（P<0.05）. The levels of serum sICAM-1 and hs-CRP in recovery phase were lower than those in acute phase（P<0.05）. The results of Pearson correlation analysis showed that serum sICAM-1 and hs-CRP levels in acute phase of children with acute viral lower respiratory tract infection were positively correlated with APACHE Ⅱ score（r values were 0.79 and 0.70,P<0.05）,and the indicators were negatively correlated to oxygenation index before treatment（r values were -0.54 and -0.46,P<0.05）. Conclusions The levels of serum sICAM-1 and hs-CRP in children with acute viral lower respiratory tract infection can reflect the development and severity of children's condition,and they can be used as reference indicators for monitoring children's condition.

Application of CLSI EP06-Ed2 in the linear verification of quantitative measurement procedures

LI Xiaobo, PU Zhifei, WU Xiaoqi, LIU Yangyang, XIE Xuanbo, SHAN Hanming, YU Jiaping

2022, 37(3):  230-234.  DOI: 10.3969/j.issn.1673-8640.2022.03.008

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Objective To investigate the application of the Clinical and Laboratory Standards Institute（CLSI） document EP06-Ed2 in the linear verification of quantitative measurement procedures,taking triiodothyronine（T₃） and free triiodothyronine（FT₃） as examples. Methods The samples close to the upper and lower limits of the linear interval were selected,7 samples were prepared proportionally,and each sample was repeated for determination twice. A weighted linear regression analysis was performed using dilution ratios and means（x）. The predicted value was calculated,and the 95% confidence interval（CI） of linear deviation was compared with the allowable deviation from linearity（ADL）. When the CI of all samples overlapped with ADL,the linearity of the whole interval was acceptable. Results The linear deviation 95%CI of samples No. 1-6 of T₃ was within the ADL,and the linear deviation of sample No. 7 was within the ADL. The linear deviation 95%CI of samples No. 2-4 of FT₃ was within the ADL,the linear deviation of samples No.1 and No.5 was within the ADL,and the linear deviation of sample No. 6 was not within the ADL,but the linear deviation 95%CI overlapped with the ADL,sample No. 7 linear deviation 95%CI was not within the ADL. Conclusions Compared with EP06-A,CLSI EP06-Ed2 has clearer and more reasonable requirements for the selection of linear sample concentration,with more effective and simple datum analysis and processing. It is a practical guide for the linear verification of quantitative measurement procedures.

Correlation of systemic inflammatory markers and fracture occurrence in elderly patients with osteoporosis

SONG Yunxiao, TONG Wei, ZHANG Haichen, ZHANG Yinwang, BIAN Xiaobo, ZHANG Linlin, YUAN Wenhua, ZHAO Zhiyun, GE Wen, YAO Tianyue

2022, 37(3):  235-239.  DOI: 10.3969/j.issn.1673-8640.2022.03.009

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Objective To investigate the correlations between systemic immune-inflammation index（SII）,platelet/lymphocyte ratio（PLR）,neutrophil/lymphocyte ratio（NLR）,lymphocyte/monocyte ratio（LMR） and C-reactive protein（CRP） and fracture in elderly patients with osteoporosis. Methods Totally,249 elderly patients with fracture（fracture group,including 108 cases of traumatic fracture and 141 cases of brittle fracture） and 177 elderly patients with osteoporosis（osteoporosis group） were enrolled. Platelet,lymphocyte,monocyte,neutrophil and CRP levels were determined,and SII,PLR,NLR and LMR were calculated. The risk factors for fracture in elderly patients with osteoporosis were assessed by binary Logistic regression analysis. Receiver operating characteristic（ROC） curves were used to analyze the value of each indicator for the diagnosis of brittle fracture and traumatic fracture alone and in combination. Results The levels of SII,PLR,NLR and CRP in fracture group were higher than those in osteoporosis group（P<0.001）,LMR levels were lower than those in osteoporosis group（P<0.001）. ROC curve analysis showed that CRP,SII,PLR,NLR,LMR,CRP determinations and combined determination were 0.589,0.899,0.835,0.866,0.788 and 0.903,respectively,for distinguishing brittle fracture from traumatic fracture. The results of binary Logistic regression analysis showed that increased SII,PLR,NLR and CRP and decreased LMR were risk factors for brittle fracture in elderly patients with osteoporosis [odds ratios（OR） were 1.001,1.008,1.178,1.021 and 0.813,95% confidence intervals（CI） were 1.000-1.001,1.005-1.011,1.084-1.279,1.013-1.028,0.744-0.888,respectively]. Conclusions SII,PLR,NLR,LMR and CRP are related to fracture occurrence in elderly patients with osteoporosis.

Establishment of the reference intervals of leukocyte surface CD64 and HLA-DR indexes for healthy adults and related factors

MIAO Linzi, LU Yao, QU Chenxue, YOU Ran, GONG Yan

2022, 37(3):  240-245.  DOI: 10.3969/j.issn.1673-8640.2022.03.010

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Objective To establish the reference intervals of leukocyte surface CD64 and HLA-DR indexes for healthy adults,and to investigate the factors on the reference intervals. Methods Totally,126 healthy subjects aged 22 to 60 years,including 69 males and 57 females,were enrolled from Peking University First Hospital. The expressions of CD64 and HLA-DR on the surface of leukocytes were determined by flow cytometry. Some related indicators were calculated through formula. The reference intervals of CD64 and HLA-DR were established. The influence of age and sex on the reference intervals was investigated. Results There was no statistical significance between different ages and CD64（P>0.05）. The mean fluorescence intensity of monocyte HLA-DR in 18-29-year-old group was lower than those in 40-49-year-old and 50-59-year-old groups（P>0.05,P=0.023,P=0.007）. The mean fluorescence intensity of monocyte HLA-DR in 18-29-year-old group was lower than that in 50-59-year-old group（P=0.034）.There was no statistical significance in leukocyte surface CD64 and HLA-DR indexes between males and females（P>0.05）. Conclusions The reference intervals of leukocyte surface CD64 and HLA-DR indexes for healthy adults have been established. Age may influence the results of leukocyte surface HLA-DR in healthy adults.

Preimplantation genetic testing of infantile polycystic kidney disease by next generation sequencing

HE Tianwen, LU Jian, CHEN Chuangqi, LIU Dun, DING Hongke, LIU Ling, DU Li, ZHENG Yichun, YIN Aihua

2022, 37(3):  257-263.  DOI: 10.3969/j.issn.1673-8640.2022.03.014

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Objective To investigate the application value of next generation sequencing technology in preimplantation genetic testing（PGT） of autosomal recessive polycystic kidney disease（ARPKD） family. Methods A case of ARPKD family was selected,and the mutation of polycystic kidney and hepatic disease 1（PKHD1） gene in family members was investigated by Sanger sequencing. The coding region of PKHD1 gene was selected as target region,120 high-density closely linked single nucleotide polymorphisms（SNP） were selected as the genetic linkage markers in the upstream and downstream 2 M regions of the gene. The SNP haplotypes of family members were constructed by selecting effective SNP sites by multiplex polymerase chain reaction（PCR） and next generation sequencing,and the risk chromosomes of gene mutation were determined. After the whole genome amplification of the trophoblast cells obtained by blastocyst biopsy,the mutation sites of embryonic PKHD1 gene were sequenced directly,and the embryonic SNP haplotypes were constructed for linkage analysis by next generation sequencing. Sanger sequencing was used to verify the next generation sequencing results of embryonic PKHD1 gene mutation sites. Low depth chromosome aneuploidy screening was carried out both in the normal and the heterozygous embryos. Results The father of the family members carried PKHD1 gene c.5935G>A,which was heterozygous. The mother carries PKHD1 gene c.10058T>G,which was heterozygous. The proband carried double heterozygous mutations of PKHD1 gene c.5935G>A and c.10058T> G. Among the 5 embryos used for biopsy,2 cases were undetected mutations,and 3 cases carried heterozygous mutations. Low depth chromosome aneuploidy screening showed that the 3 of 5 embryos were euploidy,and 2 embryos were aneuploidy. A healthy baby was delivered at full term after a well-developed euploid embryo with no mutation,which was implanted into the mother's uterus. Conclusions The application of next generation sequencing in PGT of ARPKD can block the risk of recurrence of this single gene disease in the family,and avoid the abortion caused by the selection of aneuploid embryos.

Application of morphological analysis of circulating tumor cells in clinical examination

YU Qi, SUN Yi, WANG Qiongli, CAI Yiting, LI Li

2022, 37(3):  264-269.  DOI: 10.3969/j.issn.1673-8640.2022.03.015

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Objective To investigate the application of morphological analysis of circulating tumor cells（CTC） in tumor clinical examination. Methods Peripheral blood samples of 212 patients with tumors were collected and analyzed by CTCBIOPSY detection system. The morphological analysis of CTC was performed by microscopy. The clinical applicability of morphological criteria was further verified by immunohistochemistry with CD45 and CD31 antibodies. The relationship between CTC count and detection rate and clinical factors,such as tumor type and clinical stage,was analyzed. Results The detection rate of CTC was 41.51%,and the specificity was 100%. Comparing the CTC-positive group with the CTC-negative group,there was statistical significance in age（P=0.036） and TNM stage（P<0.001）. The detection rate of CTC and CTC count were different in different types of tumors and different stages of the same type of tumor. The detection rate of CTC was the highest in patients with advanced lung cancer and prostate cancer,and the difference of CTC count in patients with early and advanced tumors had statistical significance（P<0.001）. Conclusions CTC morphological analysis can preserve the integrity of cells,and it can judge rapidly the CTC of various tumors,with good specificity and general sensitivity.

Performance comparison of lymphocyte subset determination by 6-color flow cytometry with 2 different reagents

JU Yinghui, MO Huifang, WU Hui, CHEN Pu, GUO Wei, WANG Beili

2022, 37(3):  270-273.  DOI: 10.3969/j.issn.1673-8640.2022.03.016

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Objective To compare the performance of peripheral blood lymphocyte subset determination by flow cytometry with domestic 6-color flow cytometry reagents（referred to as domestic reagents） and imported 6-color flow cytometry reagents（referred to as imported reagents）. Methods The whole blood samples from 200 patients from 3 clinical laboratories in different areas were determined by domestic reagents and imported reagents on the same flow cytometer to analyze the percentages and the absolute values of lymphocyte subsets（CD3⁺,CD4⁺,CD8⁺,CD19⁺,CD3^-CD16⁺CD56⁺ cell）. The staining indexes（SI） of lymphocyte subsets were calculated,and the deviation of the 2 reagents was evaluated. Results Compared with imported reagents,the percentages of CD3⁺ cells,CD8⁺ cells,CD19⁺ cells and the absolute values of CD3⁺ cells,CD19⁺ cells in domestic reagents had statistical significance（P<0.05）. The average deviations of the percentages and the absolute values of CD3⁺ cells,CD4⁺ cells,CD8⁺ cells,CD19⁺ cells and CD3^-CD16⁺CD56⁺ cells determined by the 2 reagents were <5%. The intraclass correlation coefficient（ICC） of the consistency was >0.95 for all the items（P<0.001）. The results of Spearman correlation analysis showed that the determination results of lymphocyte subsets of domestic reagents and imported reagents had a good correlation（r>0.9,P=0.000）. Conclusions Although the results of lymphocyte subsets with domestic reagents and imported reagents are somewhat different,the correlation and consistency are good. Each clinical laboratory can choose the appropriate antibody reagent according to its own needs.

Regulatory effect of lipid toxicity stress on Bcl-2-induced islet β cell apoptosis

ZHONG Lihong, QIU Chuanghua, PENG Ziyuan, SHE Jijia

2022, 37(3):  274-280.  DOI: 10.3969/j.issn.1673-8640.2022.03.017

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Objective To investigate the regulatory effect of lipid toxicity stress on B-cell lymphoma（Bcl）-2-induced islet β cell apoptosis. Methods C57BL/6 mice were used to establish PUMA-/- mice,and pancreatic islets were isolated after 72 h of feeding a high-fat diet. The mouse pancreatic tumor cells MIN6 were treated with palmitate and proteasome inhibitor MG132 for 0,2,4,8 and 24 h. The mouse pancreatic islet β cells were treated with sulforaphane（SFN）,palmitate and SFN + palmitate for 0,2,4,8 and 24 h. MIN6 cells and β cells treated with dimethyl sulfoxide for the same time were used as controls. The cell proteasome activity was detected,and by real-time quantitative polymerase chain reaction（PCR） the activated transcription factor 4（ATF4） mRNA,heavy-chain binding protein（Bip） mRNA,C/EBP homologous protein（Chop） mRNA,p53 up-regulated modulator of apoptosis（PUMA） mRNA expression were determined. Western blotting was used to determine the expressions of Bcl-2,Bcl-XL,Mcl-1 and Akt protein,and the cell viability at the same time was evaluated. Results Compared with 0 h,the cell proteasome activity of MIN6 cells treated with palmitate for different times did not change（P>0.05）. The proteasome activity of MIN6 cells treated with MG132 for 8 h was reduced（P<0.05）. After MG132 treated MIN6,the level of ubiquitinated protein in the cells at 24 h was increased（P<0.05）. After 24 h of treatment in palmitate group and MG132 group,the number of cell death in MIN6 was higher than that in control group（P<0.05）. The expressions of ATF4 mRNA,Bip mRNA,Chop mRNA and PUMA mRNA in MIN6 cells were higher than those of 0 h after palmitate and MG132 treatment for 24 h（P<0.05）. The protein levels of Bcl-2,Bcl-XL and Akt in MIN6 cells were lower than those of 0 h after treatment with palmitate and MG132 for 24 h（P<0.05）. The level of Mcl-1 protein increased firstly and then decreased. The level of Mcl-1 protein was the highest when treated with palmitate for 2 h,and the level of Mcl-1 was the highest when treated with MG132 for 8 h. The β cell apoptosis rate in palmitate group was higher than those in SFN group,SFN+palmitate group and control group（P<0.05）. There was no statistical significance in the β cell apoptosis rate among SFN group,SFN+palmitate group and control group（P>0.05）. AFT4 protein,Chop protein,Bip protein and PUMA mRNA in palmitate group were higher than those in control group（P<0.05）. There was no statistical significance between SFN+palmitate group and control group（P>0.05）. Conclusions Targeting ubiquitin-proteasome system （UPS） and Bcl-2 may prevent β-cell apoptosis in islet β cell effectively.

Effect of long non-coding RNA SNHG5 on the transcriptome of human hypopharyngeal carcinoma cells and clinical significance

ZHOU Huan, TANG Ying, DU Cuiping, YANG Yang, FENG Huijie, QIU Miaoxin, YAN Peiyi, CAI Xiaoyao, JIN Shu

2022, 37(3):  281-287.  DOI: 10.3969/j.issn.1673-8640.2022.03.018

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Objective To investigate the effect of long non-coding RNA（lncRNA） small nucleolar RNA host gene 5（SNHG5） on the transcriptome of human hypopharyngeal carcinoma cell line FaDu. Methods The rLV-hSNHG5-ZsGreen-Puro lentivirus and rLV-ZsGreen-Puro control lentivirus were constructed. FaDu cells infected with rLV-hSNHG5-ZsGreen-Puro lentivirus were served as rLV-SNHG5 group,and those infected with rLV-ZsGreen-Puro control lentivirus were served as rLV group. Differential gene sets were screened,and the gene transcriptional ability,Gene Ontology（GO） function,Kyoto Encyclopedia of Genes and Genomes（KEGG） signaling pathway,Reactome pathway and Disease Ontology（DO） function were analyzed. Results The relative expression of SNHG5 in rLV-SNHG5 group was higher than that in rLV group（P<0.01）. A total of 1 070 differentially expressed genes were screened by gene differential analysis,of which 537 were up-regulated,and 533 were down-regulated. The top 10 up-regulated genes with the most significant difference were LCP1,NT5E,CTGF,AXL,AKAP12,PXDN,THBS1,4-Mar,TMEM200A and RIPOR3,and the top 10 down-regulated genes were PKP1,SERPINB13,KLK10,AC011473.4,NOTCH3,CA2,EGLN3,SLC6A8,KRT4 and VAV3. Functional analysis showed that SNHG5 changed the gene transcriptional ability of some genes in FaDu cells and participated in the regulation of GO function,KEGG signaling pathway,Reactome pathway and DO function. Conclusions SNHG5 has a certain effect on the transcriptome of FaDu cells.

Research progress of clinical application of copeptin in different diseases

MAO Jie, LU Qingqing, LI Ping, SHI Shengjie, LI Jing, DENG Changjuan, TAN Chaochao, XIE Xiaobing

2022, 37(3):  291-294.  DOI: 10.3969/j.issn.1673-8640.2022.03.020

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Copeptin is the C-terminal peptide of the precursor molecule of arginine vasopressin（AVP）,which can be released into the blood equimolar with AVP by equal molar,and the half-life of it is about 14 d,which is stable and easy to detect. Copeptin has low specificity and no organ specificity,but it has high positive predictive value and high sensitivity for acute myocardial infarction（AMI）,stroke,chronic kidney disease and other diseases. This review focuses on the clinical application of copeptin in cardiovascular disease,liver disease,kidney disease,respiratory disease and stroke.