Table of Content

30 June 2024, Volume 39 Issue 6

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Determination results of serum ANA and relation with disease in 7 803 children

GAO Limei, GAO Xiupan, ZENG Junxiang, YU Youyou, PAN Xiujun

2024, 39(6):  517-523.  DOI: 10.3969/j.issn.1673-8640.2024.06.001

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Objective To investigate the distribution characteristics and disease correlation of serum antinuclear antibody（ANA） in children. Methods A total of 7 803 hospitalized children in Xinhua Hospital of Shanghai Jiao Tong University School of Medicine from January 2016 to November 2021 were enrolled. Serum ANA was determined by indirect immunofluorescence（IIF） and line immunoassay（LIA）. According to the clinical diagnosis，the patients were classified into antinuclear antibody-associated rheumatic disease（AARD） group（296 cases），other rheumatic disease（ORD） group（2 314 cases），non-rheumatic disease（NRD） group（5 128 cases） and highly suspected AARD but undetermined（UD） group（65 cases）. According to the ANA results，the patients were classified into 4 types of results，IIF-LIA-，IIF+LIA-，IIF-LIA+ and IIF+LIA+，in which IIF-LIA- was ANA negative，and the other 3 types of results were ANA positive. Results Among the 7 803 children，1 550（19.9%） cases were ANA positive. The positive rate of ANA in AARD group was the highest（81.8%），which was higher than those in ORD，NRD and UD groups（P<0.05）. The determination rate of ANA double positive（IIF+LIA+） was the highest in AARD group（48.3%），and the diagnostic likelihood ratio of ANA double positive in AARD group was 27.03. The top 3 ANA in LIA positive samples were anti-SS-A antibody [33.2%（168/506）]，anti-double-stranded DNA（dsDNA） antibody [24.1%（122/506）] and anti-histone antibody [18.0%（91/506）]. The 2 common ANA patterns in AARD group were speckled（41.9%） and homogeneous（30.7%），and high-titer ANA was the most. Receiver operating characteristic（ROC） curve results showed that the cut-off value of ANA titer was 1∶80，and the area under curve（AUC） was 0.870 2. The sensitivity was 71.3%，and the specificity was 93.5%. With the increase of ANA titers，the corresponding likelihood ratio was increased logarithmically. Conclusions Both IIF and LIA have their own advantages in the determination of ANA in children，and they are irreplaceable for each other. Double positive ANA and high titer of ANA have diagnostic value for AARD. The titer-specific likelihood ratios should be recommended as a reliable index to assess the authenticity of ANA assay.

Establishment of reference interval of serum ferritin in adults in Shanghai by indirect method

LIU Xiaowen, SHAO Wenqi, ZHU Jing, YANG Jing, ZHANG Chunyan, PAN Baishen, WANG Beili, GUO Wei

2024, 39(6):  524-529.  DOI: 10.3969/j.issn.1673-8640.2024.06.002

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Objective To establish a reference interval of serum ferritin in adults in Shanghai by indirect method. Methods A total of 27 775 individuals with ferritin determination from January 2021 to October 2022 in Zhongshan Hospital of Fudan University were enrolled. By the Turkey method to eliminate outliers，18 264 individuals were enrolled ultimately，and a reference interval of ferritin was established by the Hoffmann method. Totally，2 486 apparently healthy adults from March 2023 to April 2023 in Zhongshan Hospital of Fudan University were enrolled to validate the ferritin reference interval. The reference interval establishment population and validation population were classified into male group，female ≤50 years old group and female >50 years old group based on sex and ages. Additionally，207 females >50 years old with anemia from March 2022 to April 2023 in Zhongshan Hospital of Fudan University were enrolled to evaluate the application value of the reference interval of ferritin. The correlation between ferritin and sex，ages was evaluated by linear-regression analysis. Results There was statistical significance in ferritin levels among male group，female ≤50 years old group and female >50 years old group（P<0.001）. There was no correlation between male ferritin levels and ages（r²=0.003，P>0.05），while female ferritin levels were positively correlated with ages（r²=0.125，P<0.05）. The reference intervals were established for males，females ≤50 years old and females >50 years old，respectively，by the Hoffmann method. The reference intervals were 33.4-1 000.0 μg·L^-1 for males and 4.8-341.0 μg·L^-1 for females ≤50 years old，27.4-765.0 μg·L^-1 for females >50 years old. The upper and lower limits of the established reference intervals were different from those of the current reference intervals. The validation pass rates of established reference intervals were 98.59% for males and 95.82% for females ≤50 years old，97.38% for females >50 years old，which were higher than those of the current reference intervals（85.89%，78.76% and 60.26%，respectively）（P<0.001）. The established reference intervals can better distinguish individuals with abnormal iron metabolism indicators. Conclusions Serum ferritin exhibits sex and age differences. Serum ferritin reference intervals established by indirect method is suitable for the population in Shanghai than the current reference intervals.

Expressions of MFG-E8 and GNA14 in endometriosis associated ovarian cancer

CHENG Yan, ZHANG Ning, DONG Li, LING Xiaoming, WANG Yue, CHEN Guiqin, JING Shuang, CHEN Yannan

2024, 39(6):  530-535.  DOI: 10.3969/j.issn.1673-8640.2024.06.003

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Objective To investigate the expressions of milk fat globule-epidermal growth factor-8（MFG-E8） and G protein subunit alpha 14（GNA14） in endometriosis associated ovarian cancer（EAOC）. Methods A total of 51 patients with EAOC（EAOC group），63 patients with atypical endometriosis（AEM）（AEM group） and 82 patients with ovarian endometriosis（EMT）（EMT group） were enrolled from Fengfeng General Hospital of North China Medical and Health Group from May 2015 to May 2017. The clinical data of all the patients were collected，and the expressions of MFG-E8 and GNA14 in pathological tissues were determined. The EAOC patients were followed up for 5 years. Kaplan-Meier survival curve was used to analyze the survival of EAOC patients. Multivariate Cox regression analysis was used to evaluate the risk factors of death in EAOC patients. Results The positive rates of MFG-E8 and GNA14 in EAOC group were 82.35% and 70.59%，respectively，which were higher than those in AEM group（41.27%，44.44%） and EMT group（25.61%，23.17%）（P<0.001）. There was statistical significance in tumor diameter，tumor grade，the International Federation of Gynecology and Obstetrics（FIGO） stage and lymphatic metastasis between MFG-E8 high expression group and low expression group（P<0.05）. There was statistical significance in lymphatic metastasis between GNA14 high expression group and low expression group（P=0.009）. The progression-free survival times of MFG-E8 high expression group and GNA14 high expression group were shorter than those of MFG-E8 low expression group and GNA14 low expression group，respectively（P<0.05）. FIGO stage Ⅱ，lymphatic metastasis，MFG-E8 high expression and GNA14 high expression were independent risk factors for death in EAOC patients [hazard ratios（HR） were 2.337，2.519，3.133 and 3.080，95% confidence intervals（CI） were 1.258-4.342，1.332-4.764，1.381-7.108 and 1.318-7.197，respectively，P<0.01]. Conclusions MFG-E8 and GNA14 are highly expressed in cancer tissues of EAOC patients，and they are related to poor prognosis of patients，which may be used as prognostic indicators of EAOC patients.

Relationship between CYP2C19 polymorphism and miR-374b-5p with neurological impairment and short-term prognosis in patients with acute cerebral infarction

ZHAO Qian, ZENG Limin, ZHOU Liping, QI Lin

2024, 39(6):  536-541.  DOI: 10.3969/j.issn.1673-8640.2024.06.004

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Objective To investigate the relationship between cytochrome P450 2C19（CYP2C19） polymorphism，serum miR-374b-5p level with neurological impairment and short-term prognosis in patients with acute cerebral infarction（ACI）. Methods A total of 164 ACI patients admitted to Zhengzhou Seventh People's Hospital from January 2019 to December 2022 were enrolled. The general data of all the patients were collected，and CYP2C19 genotype and serum miR-374b-5p relative expression levels were determined. The National Institutes of Health Stroke Scale（NIHSS） was used to evaluate the degree of neurological impairment in ACI patients. According to the NIHSS score after 15 d of treatment，the ACI patients were classified into good prognosis group and poor prognosis group. Spearman correlation analysis was used to evaluate the correlation between serum miR-374b-5p and NIHSS score at admission. Multivariate Logistic regression analysis was used to evaluate the risk factors for poor short-term prognosis in ACI patients. The efficacy of CYP2C19 genotype and serum miR-374b-5p in determining the short-term prognosis of patients with ACI was evaluated by receiver operating characteristic（ROC） curve. Results Among the 164 ACI patients with CYP2C19 genotype，there were 26 cases（15.86%） of GG type（wild homozygote），66 cases（40.24%） of GA type（mutant heterozygote） and 72 cases（43.90%） of AA type（mutant homozygote）. The allele frequency of A was 64.02%，and the allele frequency of G was 35.98%. Genotype distribution was consistent with Hardy-Weinberg equilibrium law（χ²=2.620，P=0.106）. The NIHSS scores of GG genotype，GA genotype and AA genotype were increased in turn on admission（P<0.001）. The relative expression level of serum miR-374b-5p was negatively correlated with the NIHSS score on admission（r_s=-0.510，P<0.001）. The incidence of poor prognosis in ACI patients was 19.51%（32/164）. There was statistical significance in age，infarct volume，time from onset to treatment，white blood cell（WBC） count，proportion of CYP2C19 AA genotype and relative expression level of serum miR-374b-5p between good and poor prognosis groups（P<0.05）. Infarct volume，time from onset to treatment，WBC count and CYP2C19 AA genotype were all risk factors for poor prognosis in ACI patients（P<0.05），and the relative expression level of miR-374b-5p was protective factor（P=0.007）. The areas under curves（AUC） of AA genotype and serum miR-374b-5p for determining poor prognosis of ACI patients were 0.631，0.730 and 0.802，respectively. Conclusions Both CYP2C19 polymorphism and serum miR-374b-5p relative expression level are related with neurological impairment in ACI patients，which can be used as the indicators of short-term prognosis in ACI patients.

Expression of mitochondrial autophagy signaling pathway PINK1/Parkin and its relationship with neural function in patients with vascular Parkinsonism

LOU Zhan, PENG Tao, LIU Xingliang, YUE Binghong, LI Ran, ZHI Yongyi

2024, 39(6):  542-547.  DOI: 10.3969/j.issn.1673-8640.2024.06.005

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Objective To investigate the changes in serum levels of PTEN-induced putative kinase 1（PINK1）/Parkin related proteins induced by mitochondrial autophagy signaling pathway in patients with vascular Parkinsonism（VP） and its effect on neural function. Methods Totally，55 VP patients（VP group） and 55 healthy subjects（healthy control group） were enrolled from the First Affiliated Hospital of Hebei North University from March 2020 to May 2022. PINK1 and Parkin protein levels were determined. A total of 30 SD rats were selected and classified into control group（15 rats） and VP group（15 rats）. The VP rat model was constructed by intraperitoneal injection of 1-methyl-4-phenyl-1，2，3，6-tetrahydropyridine（MPTP），and the control group was injected with the same amount of 0.9% NaCl solution. For 24 h after the successful construction of the VP model，the survival rate，early apoptosis rate and late apoptosis rate of rat brain tissue were detected by flow cytometry. The mRNA and protein expressions of autophagy genes（PINK1 and Parkin） in rat brain tissue were detected by fluorescence quantitative polymerase chain reaction（PCR） and western blot. Simultaneously，the level of phosphorylation modification of PINK1/Parkin signaling pathway was detected [phosphorylation of PTEN-induced putative kinase 1（p-PINK1） protein and phosphorylation of Parkin（p-Parkin） protein]. The changes of brain ultrastructure were verified in situ by transmission electron microscopy，and the relative expression levels of autophagy proteins [autophagy-related gene（ATG）4 and ATG7] and apoptotic molecules（Bcl-2 and Bax） were detected by western blot. Results The serum PINK1 and Parkin levels in VP group were higher than those in healthy control group（P<0.01）. Compared with the control group，the survival rate of VP rats was decreased（P<0.01），and the early and late apoptosis rates were increased（P<0.01）. The relative expressions of PINK1 mRNA and Parkin mRNA were increased（P<0.01），and the relative expressions of PINK1 protein，Parkin protein，p-PINK1 protein and p-Parkin protein were increased（P<0.01）. The results of transmission electron microscopy showed that autophagy and mitochondrial autophagy were obviously damaged in VP rats. Compared with the control group，the relative expression of Bcl-2 protein in VP rats was decreased（P<0.001），and the relative expressions of Bax，ATG4 and ATG7 proteins were increased（P<0.01）. Conclusions VP can increase brain autophagy and aggravate mitochondrial autophagy damage，which is related to the overactivation of brain autophagy signaling pathway PINK1/Parkin.

Correlation of adenosine A2A receptor gene polymorphism and aggressive behavior in patients with schizophrenia

LI Sha, TAO Weiqun, HUANG Chan, CHEN Yuanhua, LAN Wenjing, LIN Shuxing

2024, 39(6):  548-556.  DOI: 10.3969/j.issn.1673-8640.2024.06.006

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Objective To investigate the correlation of adenosine A2A receptor（ADORA2A） gene single nucleotide polymorphism（SNP） and aggressive behavior in patients with schizophrenia. Methods Totally，282 patients with schizophrenia in Shenzhen Kangning Hospital from June 2015 to June 2019 were enrolled. According to the modified overt aggression scale（MOAS） scores，the subjects were classified into aggressive group（109 cases，MOAS score≥4） and non-aggressive group（173 cases，MOAS score<4）. The general data and clinical laboratory determination results were collected. ADORA2A rs5996696 and rs2298383 were determined. Logistic regression analysis was used to analyze the risk factors and the interaction with ADORA2A gene SNP. The nomogram prediction model was established and verified. Results In aggressive group，the proportion of males，history of alcohol consumption，positive and negative syndrome scale（PANSS） score，involuntary hospitalization，previous aggressive behavior，unemployment，recent severe stress events，elevated rate of total triiodothyronine（TT₃），interleukin-17（IL-17），interleukin-23（IL-23） and ADORA2A rs2298383 CC genotype frequency were higher than those in non-aggressive group（P<0.05）. PANSS score≥20，IL-17≥16.13 pg·mL^-1，elevated TT₃，previous aggressive behavior and carrying ADORA2A rs2298383 CC genotype were independent risk factors for aggressive behavior in schizophrenia patients（P<0.05）. CC genotype of ADORA2A rs2298383 had positive interaction with PANSS score，elevated TT₃，IL-17 level and previous aggressive behavior. The total risk value of aggressive behavior predicted by the nomogram prediction model was 0.93，the area under receiver operating characteristic （ROC）curve（AUC） was 0.874，and there was no statistical significance in the goodness of fit test of the calibration curve（P>0.05）. Conclusions ADORA2A rs2298383 CC genotype is correlated with aggressive behavior in schizophrenia patients，and which is positively interacted with risk factors associated with aggressive behavior in schizophrenia patients.

Clinical value of inflammatory derived indicators and cystatin C in preeclampsia

CHEN Haiying, GE Yafang, PAN Shun, YANG Haiou

2024, 39(6):  557-561.  DOI: 10.3969/j.issn.1673-8640.2024.06.007

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Objective To investigate the clinical value of inflammatory derived indicators [derived neutrophil-to-lymphocyte ratio（dNLR） and systemic inflammatory response index（SIRI）] and cystatin C（Cys C） in the prediction of preeclampsia. Methods From January 2019 to November 2022，194 patients with preeclampsia in the International Peace Maternity and Child Health Hospital affiliated to Shanghai Jiao Tong University School of Medicine were enrolled as observation group. A total of 152 healthy pregnant females were enrolled as control group. The relationship between dNLR，SIRI，Cys C and preeclampsia was analyzed. Results The level of Cys C in observation group was higher than that in control group（P<0.01）. The white blood cell count，neutrophil count，monocyte count，dNLR and SIRI in observation group were higher than those in control group（P<0.01）. There was no statistical significance in lymphocyte count，platelet count and mean platelet volume between observation group and control group（P>0.05）. Cys C and dNLR were positively correlated with systolic blood pressure and diastolic blood pressure，respectively（P<0.01）. SIRI was positively correlated with diastolic blood pressure（P<0.05）. The combined determination of Cys C，dNLR and SIRI predicted preeclampsia with an area under curve of 0.852 [95% confidence interval（CI） 0.812-0.892，P<0.01]，and the sensitivity and specificity were 61.8% and 92.7%，respectively. Cys C≥0.905 mg·L^-1，dNLR≥2.950，SIRI≥2.315 were independent risk factors for preeclampsia（P<0.05）. The observation group was sub-classified according to the optimal cut-off values of Cys C，dNLR and SIRI. There was no statistical significance in pregnancy outcome between high level groups and low level groups（P>0.05）. Conclusions Cys C，dNLR and SIRI are independent risk factors for preeclampsia，and they have good sensitivity and specificity，which have certain value for prediction and evaluation of preeclampsia.

Correlation between MTHFR C677T polymorphism and insulin resistance in patients with polycystic ovary syndrome

ZHAO Ying, LU Loukaiyi, DAI Sijia, CHEN Yisheng

2024, 39(6):  562-567.  DOI: 10.3969/j.issn.1673-8640.2024.06.008

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Objective To investigate the correlation between methylene tetrahydrofolate reductase（MTHFR） C677T polymorphism and insulin resistance（IR） in patients with polycystic ovary syndrome（PCOS）. Methods A total of 145 PCOS patients and 87 healthy females from the Obstetrics and Gynecology Hospital of Fudan University were enrolled from January 2022 to November 2023. The C677T polymorphism of MTHFR gene，fasting glucose（FPG） and fasting insulin（FINS） levels were determined. Homocysteine（Hcy），total cholesterol（TC），triglyceride（TG），folic acid，vitamin B₁₂（Vit B₁₂） and vitamin D（Vit D） levels were determined. Insulin release test（IRT） was also performed in patients with PCOS. Insulin sensitivity index（ISI），homeostasis model assessment for insulin resistance（HOMA-IR） and the area under insulin curve（AUCINS） were calculated. The research subjects were classified into IR and non-IR groups according to whether IR occurred or not. Results No statistical significance was found between MTHFR C677T and allele frequency distribution and between IR and non-IR groups in healthy control group（P>0.05）. The frequencies of CT，TT genotypes and T allele were higher in IR group than those in non-IR group（P<0.05）. Differences in FINS，FPG，Hcy，TG，ISI and AUCINS were found between IR and non-IR groups（P<0.05），and the other index differences were not found between the 2 groups（P>0.05）. The TC levels of CT and TT genotypes in PCOS group were higher than CC genotype（P<0.05），and the Hcy level of TT genotype were higher than CC and CT genotypes（P<0.05）. There was no statistical significance for all the genotypes in IR group（P>0.05）. Conclusions MTHFR gene C677T polymorphism is correlated with IR in females with PCOS. In PCOS patients without IR，Hcy and TC levels are increased in patients with TT genotype.

Prognosis evaluation model of children with Mycoplasma pneumoniae pneumonia based on MP DNA and peripheral blood inflammation indicators

DAI Yao, LI Jiahui, XU Xiuhong

2024, 39(6):  568-572.  DOI: 10.3969/j.issn.1673-8640.2024.06.009

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Objective To evaluate the clinical value of a model for the prognosis of children with Mycoplasma pneumoniae（MP） pneumonia（MPP） based on MP DNA and peripheral blood inflammation indicators. Methods Totally，139 children with MMP were enrolled from Jinshan Hospital of Fudan University from November to December 2023. The general clinical data were collected，and MP DNA，C-reactive protein（CRP），procalcitonin（PCT） and blood routine test indicators were determined before treatment. Neutrophil-to-lymphocyte ratio（NLR），platelet-to-lymphocyte ratio（PLR） and monocyte-to-lymphocyte ratio（MLR） were calculated. According to the prognosis after 1 month of treatment，they were classified into poor prognosis group（33 cases） and good prognosis group（106 cases）. Spearman correlation analysis was used to evaluate the correlation between various indicators and poor prognosis of MPP. Multivariate Logistic regression analysis was used to evaluate the influence factors of poor prognosis in children with MPP. Receiver operating characteristic（ROC） curve was used to evaluate the efficacy of single and combined determinations of each indicator in determining poor prognosis of children with MPP. Results The levels of MP DNA，CRP，NLR and PLR in poor prognosis group were higher than those in good prognosis group（P<0.05），but there was no statistical significance in PCT and MLR between the 2 groups（P>0.05）. MP DNA，CRP，NLR and PLR were positively correlated with poor prognosis in MPP children（r values were 0.313，0.225，0.277 and 0.262，P<0.05）. MP DNA，CRP，NLR and PLR were all risk factors for poor prognosis in MPP children（P<0.05）. The areas under curves（AUC） of single determinations of MP DNA，CRP，NLR and PLR and combined determination to determine the poor prognosis of MPP children were 0.799，0.724，0.775，0.732 and 0.935，respectively. Conclusions Both MP DNA and inflammation indicators of MPP children show abnormal changes before treatment，and the prognosis evaluation model has good clinical application value.

Changes and clinical significance of procalcitonin and coagulation function indicators in children with sepsis

JIANG Jingjing, GU Ping, WANG Chengyun, ZHANG Fan, SHEN Huiying, PAN Qiuhui, WANG Jing

2024, 39(6):  573-577.  DOI: 10.3969/j.issn.1673-8640.2024.06.010

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Objective To investigate the changes of procalcitonin（PCT） and coagulation function indicators in children with sepsis and their correlation. Methods A total of 157 children with sepsis and 132 children without sepsis admitted to Shanghai Children's Medical Center from June 2021 to June 2022 were enrolled. Relevant basic information，primary disease，the incidence of disseminated or diffuse intravascular coagulation（DIC），the incidence of shock，sequential organ failure assessment（SOFA） score，the 28-day treatment outcomes of patients were recorded. PCT and coagulation function indicators [prothrombin time（PT），activated partial thromboplastin time（APTT），fibrinogen（Fib），D-dimer（DD）] were determined. According to the levels of PCT，the sepsis group was classified into 3 groups，including Group A（PCT<2.00 ng·mL^-1），Group B（2.00 ng·mL^-1≤ PCT <10.00 ng·mL^-1） and Group C（PCT≥10.00 ng·mL^-1）. The differences of each indicator between the 3 groups of sepsis group and non-sepsis group were compared，and the correlation between different PCT levels and coagulation function indicators in sepsis group was analyzed. Receiver operating characteristic（ROC） curve was used to evaluate the efficiency of each indicator for the poor prognosis of children with sepsis. Results PT and APTT were prolonged in sepsis group，which had statistical significance with those in non-sepsis group（P<0.05）. In sepsis group，the levels of PCT，Fib and DD were higher than those in non-sepsis group（P<0.05）. The levels of PT，APTT，Fib and DD in Group A，Group B and Group C were gradually increased. The higher the PCT and the more severe sepsis were in Group A，B and C，the more patients with poor prognosis. In sepsis group，PT，APTT, Fib and DD were positively correlated with PCT level（r=0.220、0.502、0.186 and 0.166，P<0.05）. The areas under curves of PCT，PT，APTT，Fib and DD were 0.822，0.819，0.524，0.541 and 0.879，respectively. The area under curve of PCT combined with PT，APTT，Fib and DD was 0.909，which had good sensitivity（89.5%） and specificity（78.9%）. Conclusions There is a correlation between PT，APTT，Fib，DD and PCT in children with sepsis. The efficiency of dynamic combined monitoring of PCT and coagulation function indicators for the diagnosis and prognosis of sepsis is better than that of single indicator，which can provide a reference for the determination and prognosis of clinical diagnosis and treatment plan for children with sepsis.

Analysis of 4 blood culture bottles' adsorption capacity to 4 commonly used antibiotics

YANG Huilin, CHEN Juan, YAN Jinjin, OU Jiawen, WEN Mingming, ZHOU Lina

2024, 39(6):  578-582.  DOI: 10.3969/j.issn.1673-8640.2024.06.011

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Objective To analyze the adsorption capacity of 4 kinds of blood culture bottles to imipenem，piperacillin-tazobactam，vancomycin and caspofungin，and to provide a reference for improving the determination rate of pathogens in blood culture. Methods Four antibiotics（imipenem，piperacillin-tazobactam，vancomycin and caspofungin） paired with sensitive and standard isolates were used，and blood culture was performed in 4 types of blood culture bottles（FA plus，FN plus，Aerobic and Anaerobic）. The adsorption capacity of different blood culture bottles to 4 antibiotics was evaluated according to the positive situation and positive alarm time of blood culture bottles. Results Control bottles（without antibiotic adsorption measures） showed no positivity after 5 d culture. The positive alarm rates were 75.0%（15/20） on FA plus，100.0%（10/10） on FN plus，70.0%（14/20） on Aerobic and 0.0%（0/10） on Anaerobic. Only FN plus showed positivity in imipenem/Escherichia coli group，and the positive alarm time was （12.4±0.3）h. FA plus，Aerobic and Anaerobic showed no positivity for 5 d. In piperacillin-tazobactam/Pseudomonas aeruginosa group，FA plus and Aerobic showed positivity（5/5），and the positive alarm times were 16.2±0.3 and （18.1±0.4）h，respectively（P<0.05）. FA plus，FN plus and Aerobic reported positivity in vancomycin/Staphylococcus aureus group，and the positive alarm times were 16.7±0.4，24.1±0.6 and （31.3±3.9）h，respectively（P<0.05），and Anaerobic showed no positivity for 5 d. FA plus and Aerobic reported positivity in caspofungin/Candida albicans group，and the positive alarm times were 47.1±3.3 and （42.9±2.0）h（P<0.05）. Conclusions FN plus has the capacity to adsorb imipenem and vancomycin，while FA plus and Aerobic have the capacity to adsorb vancomycin，piperacillin-tazobactam and caspofungin. For patients who have used antibiotics before blood sample collection，it is not recommended to use blood culture bottles without antibiotic adsorption measures，and appropriate blood culture bottles should be selected according to clinical antibiotic use habits.

Clinical characteristics of acute lymphoblastic leukemia children infected with Epstein-Barr virus

YAN Jianghong, WANG Le, MA Lin, YANG Shuo, GUO Weiwei, ZHAO Mengchuan, LIU Zehao, ZHAI Xiaoying

2024, 39(6):  583-586.  DOI: 10.3969/j.issn.1673-8640.2024.06.012

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Objective To investigate the clinical characteristics of children with acute lymphoblastic leukemia（ALL） infected by Epstein-Barr（EB） virus，and to provide a reference for clinical intervention. Method A total of 90 ALL children aged ≤17 years hospitalized in Hebei Children's Hospital from January 2020 to August 2022 were enrolled and classified into infected group（78 cases） and uninfected group（12 cases） according to whether they were infected with EB virus. The clinical data of 2 groups of children were collected. Serum EB virus antibody profile [anti-EB virus capsid antibody IgM（EBV-CAIgM），anti-EB virus early antibody IgG（EBV-EAIgG），anti-EB virus capsid antibody IgG（EBV-CAIgG），anti-EB virus nuclear antibody IgG（EBV-NA₁IgG）] was determined by enzyme-linked immunosorbent assay. The clinical characteristics and various indexes of the 2 groups were compared. Results There was no statistical significance in gender and age between infected and uninfected groups （P>0.05）. Among children with T-acute lymphoblastic leukemia （T-ALL），70.59% （12/17） cases were infected with EB virus，while among children with B-acute lymphoblastic leukemia （B-ALL），90.41%（66/73） cases were infected with EB virus. The difference in the proportion of EB virus infection between children with different immunophenotypes was not statistically significant （P>0.05）. The proportions of previous infections among children with B-ALL and T-ALL were high （>45%）. Conclusions The proportion of EB virus infection in children with ALL is high. Clinical screening and monitoring of EB virus infection in ALL patients should be strengthened.

Clinical application of MALDI-TOF direct droplet culture method in rapid determination of CRKP

WU Xinzhe, MAO Haifeng, YANG Jin, ZUO Chunlei, JIN Danting

2024, 39(6):  587-591.  DOI: 10.3969/j.issn.1673-8640.2024.06.013

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Objective To investigate the accuracy and clinical application role of direct-on-target microdroplet growth assay（DOT-MGA） based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry（MALDI-TOF MS） for rapid determination of carbapenem-resistant Klebsiella pneumoniae（CRKP）. Methods A total of 251 clinical Klebsiella pneumoniae isolates were collected from the First Hospital of Lianyungang from 2020 to 2021，which included 123 meropenem-sensitive isolates and 128 CRKP isolates. The carbapenem resistance of all the isolates was determined by DOT-MGA with 2 sample sizes of 4 and 6 μL，and the minimum inhibitory concentration（MIC） of meropenem of each isolate was determined by broth microdilution method. Using the results of broth microdilution method as reference，the accuracy of DOT-MGA drug resistance results was evaluated. Results With the broth microdilution method as reference，the sensitivity and negative predictive value of DOT-MGA were 100.0% after incubation and growth for 4 h，and the specificity and positive predictive value of 4 μL group were 92.5% and 92.8%，respectively. The specificity and positive predictive value of 6 μL group were 89.8% and 90.1%，respectively. The MIC₅₀ of 123 meropenem-sensitive isolates was 0.125 μg·mL^-1，and the MIC₉₀ was 0.25 μg·mL^-1. The MIC₅₀ of 128 meropenem-resistant isolates was ≥128 μg·mL^-1，and the MIC₉₀ was ≥128 μg·mL^-1 as well. The consistency analysis results of DOT-MGA and broth microdilution method showed that the Kappa value of 4 μL group was 0.88，and that of 6 μL group was 0.83. Conclusions DOT-MGA based on MALDI-TOF is a simple，rapid and accurate method for the determination of CRKP，which can be used in the rapid determination of CRKP in clinical practice.

Research progress and challenges on CRISPR/Cas nucleic acid diagnosis system

ZHOU Jian, REN Xuemei, WANG Xin, LI Zhuo

2024, 39(6):  608-614.  DOI: 10.3969/j.issn.1673-8640.2024.06.017

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Clustered regularly interspaced short palindromic repeat（CRISPR）/clustered regularly interspaced short palindromic repeat-associated protein（Cas）nucleic acid diagnosis system，as a new molecular diagnosis technology has advantages of rapid determination，simple operation and high specificity. CRISPR/Cas has been a research hotspot in nucleic acid molecular diagnosis. Some CRISPR/Cas nucleic acid diagnosis systems have been validated by clinical samples，which are expected to be an ideal nucleic acid molecular diagnosis technology. However，there are still a series of problems to be solved before the clinical large-scale commercial application. In this review，the principle，characteristics and problems are summarized，and it is expected to provide a reference for the early clinical application of CRISPR/Cas nucleic acid diagnosis system.

Research progress on determination and treatment of carbapenem-resistant Klebsiella pneumoniae

SUN Miaoli, WU Qiong, WANG Yingzhi, WANG Jianqiang, GAO Feng, TANG Jin

2024, 39(6):  615-620.  DOI: 10.3969/j.issn.1673-8640.2024.06.018

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With the increasing determination rate of carbapenem-resistant Klebsiella pneumoniae（CRKP），strong CRKP drug resistance，rapid dissemination and high patients' mortality have brought challenges to clinical treatment. At present，the determination methods of CRKP mainly include carbapenem inactivation method（CIM），Carba NP test，mass spectrometry，enzymic immunochromatography and molecular determination technology. Antibiotic therapy，bacteriophage combined with antibiotics，vaccines and monoclonal antibodies are the main methods to treat CRKP.