Table of Content

30 May 2023, Volume 38 Issue 5

Previous Issue    Next Issue

Analysis on the difference of lipoprotein（a） particle concentration and mass concentration in certain disease states

WANG Yang, MO Huifang, SHEN Ruojian, SHEN Yifeng, WANG Beili, PAN Baishen, GUO Wei

2023, 38(5):  405-412.  DOI: 10.3969/j.issn.1673-8640.2023.05.001

Asbtract ( 127 )   HTML ( 15)   PDF (1991KB) ( 135 )  

Figures and Tables | References | Related Articles | Metrics

Objective To analyze the results of lipoprotein（a）[Lp（a）] mass concentration and particle concentration in certain disease states. Methods A total of 581 inpatients from September 2018 to August 2019 in Zhongshan Hospital of Fudan University were enrolled，which included 406 males and 175 females，aged （63.59±14.52） years old. There were 133 cases in coronary heart disease group，76 cases in hepatocarcinoma group，192 cases in diabetes group and 180 cases in chronic kidney disease group. Totally，200 healthy subjects were enrolled as well，which included 125 males and 75 females，aged （61.84±5.50） years old. The particle concentration and mass concentration of Lp（a） were determined，and the correlation and positive rate difference between particle and mass concentration in disease groups and healthy control group were analyzed by Spearman correlation analysis. Passing-Bablok regression analysis and Bland-Altman bias evaluation were used to analyze the consistency between the mass concentration calculated by particle concentration and the mass concentration determined directly. Results The correlation between Lp（a） particle concentration and mass concentration was good（r=0.948 7，P<0.05），the highest in coronary heart disease group（r=0.976 3，P<0.05），and the lowest in hepatocarcinoma group（r=0.863 2，P<0.05）. The positive rates of particle concentration and mass concentration in chronic kidney disease group and diabetes group had statistical significance（P<0.05）. There was no statistical significance in the positive rates of particle concentration and mass concentration among coronary heart disease group，hepatocarcinoma group and healthy control group（P>0.05）. The 95% agreement margin between mass concentration calculated by particle concentration and mass concentration determined directly exceeded 1/2 of the allowable total error. Conclusions There is a certain correlation between the particle concentration of Lp（a） and its mass concentration. In different disease groups（coronary heart disease，hepatocarcinoma，diabetes and chronic kidney disease） and healthy control group，the positive rates between Lp（a） particle and mass concentrations are different. The difference between the mass concentration calculated by particle concentration and mass concentration determined directly exceeds the clinically acceptable range，and it is not recommended to convert the Lp（a） particle concentration and mass concentration in clinical practice.

Expression levels of serum miR-4443 and TRAF4 in patients with hemorrhagic stroke

ZHAO Jing, WANG Pei, LU Dandan, QI Dandan, LI Xuan

2023, 38(5):  413-418.  DOI: 10.3969/j.issn.1673-8640.2023.05.002

Asbtract ( 74 )   HTML ( 13)   PDF (951KB) ( 49 )  

Figures and Tables | References | Related Articles | Metrics

Objective To investigate the changes of serum miR-4443 and tumor necrosis factor receptor-associated factor（TRAF） 4 expression levels in patients with hemorrhagic stroke，and to analyze the clinical value. Methods Totally，77 patients admitted to Baoding First Central Hospital from June to December 2020 who were diagnosed as hemorrhagic stroke were enrolled as stroke group. The National Institute of Health Stroke Scale（NIHSS） score was used to evaluate the degree of neurological impairment，the 77 patients were classified into mild group（<6，29 cases），moderate group（6-13，27 cases） and severe group（>13，21 cases）. The stroke group was followed up for 6 months，and the good prognosis group（32 cases） and poor prognosis group（45 cases） were established according to the prognosis of patients. Another 48 healthy subjects were enrolled as healthy control group. Pearson correlation coefficient was used to analyze the correlation between miR-4443，TRAF4 levels and the NIHSS score of patients with hemorrhagic stroke. Receiver operating characteristic（ROC） curve was used to analyze the predictive values of miR-4443，TRAF4 and the combined determination for the prognosis of patients with hemorrhagic stroke. Results Compared with healthy control group，the expression level of miR-4443 in stroke group was decreased（P<0.01），and the expression level of TRAF4 was increased（P<0.01）. The expression level of miR-4443 in stroke group was decreased with the severity of disease（P<0.001），while the expression level of TRAF4 was increased with the severity of disease（P<0.001）. Pearson correlation coefficient analysis showed that NIHSS score was negatively correlated with miR-4443 expression level（r=-0.857，P<0.001），and it was positively correlated with TRAF4 expression level（r=0.805，P<0.001）. The expression level of miR-4443 in poor prognosis group was decreased compared with that in good prognosis group（P<0.05），and the expression level of TRAF4 was increased compared with that in good prognosis group（P<0.05）. ROC curve analysis showed that the area under curve（AUC） of combined determination（0.964） was increased than those of single determinations （0.686 and 0.790）（P<0.05）. Conclusions The development degree of hemorrhagic stroke is related with the expression levels of miR-4443 and TRAF4，which have a value for the prognosis of patients.

Effect of CYP3A4 *1G polymorphism on fentanyl in patients undergoing laparoscopic radical resection of colorectal cancer

SU Xi, ZHOU Yan, HUANG Jian, GAO Yaoyi, LI Yihao, WANG Beili, PAN Baishen, GUO Wei

2023, 38(5):  419-423.  DOI: 10.3969/j.issn.1673-8640.2023.05.003

Asbtract ( 63 )   HTML ( 7)   PDF (868KB) ( 35 )  

Figures and Tables | References | Related Articles | Metrics

Objective To assess the effect of CYP3A4 *1G polymorphism on the dose of fentanyl in patients undergoing laparoscopic radical resection of colorectal cancer. Methods Totally，101 patients receiving fentanyl anesthesia laparoscopic radical resection of colon tumors at Zhongshan Hospital of Fudan University between July 2018 and April 2020 were enrolled. CYP3A4 *1G genotype was determined，and the relation between CYP3A4 *1G polymorphism and the dose of fentanyl was evaluated by multiple linear regression analysis. Results The results of multiple linear regression analysis showed that age，intraoperative fentanyl dose and the polymorphisms of CYP3A4 *1G，CYP3A5 *3 and OPRM1 A118G were risk factors for the dose of fentanyl in post-anesthesia care unit （PACU）（P<0.05）. After adjusting for age，sex，weight，height，intraoperative fentanyl dose，operation time and the polymorphisms of OPRM1 A118G，CYP3A5 *3 and COMT V158M，CYP3A4*1G wild type（*1*1） was found to be a risk factor for increased fentanyl dose in PACU [β=-13.99，95% confidence interval（CI） -26.78--1.20，P=0.035]. Compared to fentanyl dose in patients with CYP3A4 *1G mutant genotypes [*1*1G（heterozygous） and *1G*1G（homozygous）]，the wild type ones were 13.99 μg higher. There was no correlation between CYP3A4 *1G polymorphism and fentanyl dose in 24 h patient-controlled intravenous analgesia (PCIA) after operation（β=-7.79，95% CI -33.70--18.11，P=0.557），as well as that in 48 h PCIA after operation（β=-10.28，95% CI -22.70-2.15，P=0.108）. Compared with CYP3A4*1G wild type （*1*1） patients，the PACU and fentanyl dose in 24 h PCIA after operation of patients with CYP3A4*1G mutant genotypes [*1*1G （heterozygous） and *1G*1G （homozygous）] were decreased （P<0.05）. Conclusions CYP3A4 *1G polymorphism is an independent factor for fentanyl dose. Compared with mutated type，the anesthetic effect may be similar to wild type subjects by consuming less fentanyl.

Changes of serum HAS2 and CD44 levels in breast cancer patients and their clinical significance

ZHANG Guoliang, LIU Yiwen, HE Yiqing, XU Jing, YANG Cuixia, GAO Feng, LIU Hua

2023, 38(5):  424-429.  DOI: 10.3969/j.issn.1673-8640.2023.05.004

Asbtract ( 84 )   HTML ( 13)   PDF (1041KB) ( 61 )  

Figures and Tables | References | Related Articles | Metrics

Objective To investigate the changes of serum hyaluronan synthase 2（HAS2） and differentiated antigen cluster 44（CD44）levels and the clinical values in patients with breast cancer. Methods From January 2020 to June 2021，52 patients with breast cancer（breast cancer group），40 patients with benign breast diseases（benign disease group）and 34 healthy subjects（healthy control group）from Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine were enrolled. The clinical data were collected，and serum HAS2 and CD44 levels were determined. Pearson correlation analysis was used to evaluate the correlation between serum HAS2 and CD44 levels. Receiver operating characteristic（ROC） curve was used to evaluate the efficacy of single and combined determinations of serum HAS2，CD44，carcinoembryonic antigen（CEA） and carbohydrate antigen（CA）15-3 in the diagnosis of breast cancer. Results Serum HAS2 and CD44 in healthy control group，benign disease group and breast cancer group were increased successively（P<0.000 1）. Pearson correlation analysis showed that serum HAS2 and CD44 levels were positively correlated in healthy control group，benign disease group and breast cancer group（r values were 0.364 4，0.415 2 and 0.290 5，respectively，P<0.05）. ROC curve analysis showed that the areas under curves（AUC） of serum HAS2，CD44，CEA and CA15-3 by single and combined determinations in the diagnosis of breast cancer were 0.800，0.805，0.560，0.621 and 0.883，respectively. The AUC of combined determination in the diagnosis of early stage（TNM stage Ⅰ to Ⅱ） and late stage（TNM stage Ⅲ to Ⅳ）were 0.860 and 0.919，respectively. There was statistical significance in serum HAS2 level among breast cancer patients with different tumor sizes，histological grades，human epidermal growth factor receptor 2（HER2） expressions and lymphatic metastasis（P<0.05）. There was no statistical significance in serum HAS2 level among breast cancer patients with different ages，estrogen receptor（ER） expressions，progesterone receptor（PR） expressions，Ki67 expressions and vascular invasions（P>0.05）. There was no statistical significance in serum CD44 level among breast cancer patients with different clinicopathologic characteristics（P>0.05）. The AUC of serum HAS2 in the diagnosis of lymphatic metastasis of breast cancer was 0.739. Conclusions Serum HAS2 and CD44 are potential as auxiliary diagnostic indicators for breast cancer. Serum HAS2 level is related to breast cancer malignant progression，which may be used as one of the prognostic indicators.

Influence of particle radiotherapy on bone marrow，liver and kidney functions in patients with prostate cancer

YU Qi, WANG Yanchun, GUO Lin, LU Renquan

2023, 38(5):  430-434.  DOI: 10.3969/j.issn.1673-8640.2023.05.005

Asbtract ( 85 )   HTML ( 7)   PDF (887KB) ( 52 )  

Figures and Tables | References | Related Articles | Metrics

Objective To investigate the influence of particle radiotherapy on bone marrow，liver and kidney functions in prostate cancer patients. Methods A total of 23 prostate cancer patients undergoing photon radiation（photon radiation group），25 prostate cancer patients undergoing heavy ion radiation（heavy ion radiation group），28 prostate cancer patients undergoing proton heavy ion radiation（proton heavy ion radiation group） were enrolled，and according to treatment plans，each group was classified into simple radiotherapy and synchronous endocrine therapy subgroups. White blood cell（WBC） count，the absolute value of lymphocytes（LYMPH#），hemoglobin（Hb） and platelet（PLT） count were determined before and after treatment for all prostate cancer patients，in order to reflect bone marrow function suppression. The levels of albumin（Alb），total bilirubin（TB），alanine aminotransferase（ALT），aspartate aminotransferase（AST），creatinine（Cr） and urea were determined before treatment. Common Terminology Criteria for Adverse Events（CTCAE） version 5.0 was used. The liver and kidney functions before and after treatment in 15 patients undergoing synchronous endocrine therapy and photon radiation and 13 patients with synchronous endocrine therapy were determined and compared. Results There was no statistical significance in WBC count，Hb，PLT count，TB，ALT and urea in photon radiation group，heavy ion radiation group and proton heavy ion radiation group before treatment（P>0.05）. There was statistical significance in Alb，AST，Cr and LYMPH# among the 3 groups（P<0.05）. The proportion of LYMPH # grade Ⅰ patients after heavy ion radiation was higher（100%，12/12） than those after photon radiation（50%，4/8） and proton heavy ion radiation（50%，4/8）（P=0.015）. The proportion of patients with LYMPH# grade Ⅰ after synchronous endocrine therapy of heavy ion radiation（100%，13/13） was higher than that of photon radiation（33%，5/13） and proton heavy ion radiation（35%，7/20）（P=0.002）. After the synchronous endocrine therapy of heavy ion radiation and proton heavy ion radiation，the proportions of Hb grade Ⅰ were 100%（13/13，20/20），which was higher than that of photon radiation（73%，11/15）（P=0.048）. After photon radiation combined with endocrine therapy，serum ALT level was higher than that before treatment（P=0.005），and serum Cr level was lower than that before treatment（P=0.026）. There was no statistical significance in serum Alb，TB，AST and urea levels before and after treatment（P>0.05）. There was no statistical significance in Alb，TB，ALT，AST，Cr and urea before and after heavy ion radiation combined with endocrine therapy（P>0.05）. Conclusions The degree of bone marrow function suppression of heavy ion radiation is lower than that of photon radiation and proton heavy ion radiation，whether it is synchronous endocrine therapy or not. The suppression effect on liver and kidney functions is lower than that of photon radiation combined with endocrine therapy.

Classification and epidemiology characteristics of Klebsiella pneumoniae isolated from clinic

ZHU Yurong, ZHANG Dan, HE Yaxing, LI Jingjing, HUANG Jing, LI Ting, LIU Peng, LIU Ronghua

2023, 38(5):  435-440.  DOI: 10.3969/j.issn.1673-8640.2023.05.006

Asbtract ( 94 )   HTML ( 7)   PDF (1002KB) ( 57 )  

Figures and Tables | References | Related Articles | Metrics

Objective To analyze the classification and epidemiology characteristics（capsular serotype，drug resistance gene and virulence gene） of Klebsiella pneumoniae isolated from clinic，and to provide a reference for diagnosis and treatment. Methods Non-repetitive 143 isolates of Klebsiella pneumoniae were collected from Linfen Central Hospital from January 2021 to January 2022. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was used for isolate identification，and the drug resistance was determined by VITEK-2 system and K-B disk diffusion. Molecular classification and capsular serotype were performed by polymerase chain reaction（PCR） and gene sequencing. Virulence genes and drug resistance genes were determined by polymerase chain reaction（PCR）. Results A total of 4 isolates of Klebsiella variicola（KpⅢ），4 isolates of Klebsiella quasipneumoniae（KpⅡ） and 135 isolates of Klebsiella pneumoniae（KpⅠ） were identified. The main capsular serotypes of KpⅠ were K1，K2 and K57. K1 was the main capsular serotype of KpⅡ. The capsular serotypes of KpⅢ dispersed distribution. KpⅢ and KpⅡonly carried with SHV，OKP and LEN genes，while KpⅠcarried multiple drug resistance genes. The drug resistance of KpⅠwas higher than others. Virulence genes were determined in all the isolates. The rate of virulence genes for fimH，mrkD and ENT was >95% in the 3 isolate groups. The carrying rates of KFU in KpⅡ and KpⅢ（100.00%） were higher than that in KpⅠ（73.33%），and the carrying rate of YBT in KpⅠ，KpⅡ and KpⅢ were 54.81%，25.00% and 0%，respectively. Conclusions The clinical isolates of Klebsiella pneumoniae are mainly KpⅠ. There were differences in carrying rate of virulence gene，drug resistance gene and capsular serotype distribution.

Antibacterial activity of 4 Chinese herbal medicines alone and combined with western medicines for extensively drug resistant Acinetobacter baumannii

MA Dongmei, JIANG Dongkui, XIANG Xiaobo, LI Shiliang

2023, 38(5):  441-445.  DOI: 10.3969/j.issn.1673-8640.2023.05.007

Asbtract ( 132 )   HTML ( 4)   PDF (915KB) ( 14 )  

Figures and Tables | References | Related Articles | Metrics

Objective To analyze the antibacterial effects of Radix isatidis，Forsythia，Scutellaria baicalensis and Dandelion and combined with cefoperazone-sulbactam（SCF） for extensively drug-resistant Acinetobacter baumannii（XDR-AB）. Methods Totally，30 isolates of XDR-AB were collected from clinical specimens from September 2018 to October 2019 in the Third Affiliated Hospital of Beijing University of Chinese Medicine. The minimum inhibitory concentrations（MIC） of Radix isatidis，Forsythia，Scutellaria baicalensis and Dandelion alone and combined with SCF to 30 isolates of XDR-AB were determined by broth microdilution method. Fractional inhibitory concentrations（FIC） were calculated. The changes of K⁺ and alkaline phosphatase（ALP）levels of 4 Chinese herbal medicines were compared. Results The 4 Chinese herbal medicines have certain antibacterial effects. The MIC of Forsythia and Scutellaria baicalensis were low. The effect of Radix isatidis combined with SCF was good，and synergistic and additive effects were predominant. The effects of Forsythia and Scutellaria baicalensis combined with SCF were mainly antagonism（FIC index >2）. The K⁺ levels of the 4 groups were not statistically significant（P>0.05）. The ALP levels in Radix isatidis and Dandelion groups were not statistically significant（P<0.05），and those in Forsythia and Scutellaria baicalensis groups were not statistically significant（P>0.05）. Conclusions The antibacterial effects of Forsythia and Scutellaria baicalensis are good，the combined antibacterial effect of Radix isatidis and SCF is opitmal.

Establishment and performance evaluation of UPLC-MS/MS for 6 markers in dried blood spots

LI Xijiu, ZHOU Jianwei, ZHANG Qin, LONG Yunlin, WU Yingsong

2023, 38(5):  446-453.  DOI: 10.3969/j.issn.1673-8640.2023.05.008

Asbtract ( 69 )   HTML ( 9)   PDF (968KB) ( 82 )  

Figures and Tables | References | Related Articles | Metrics

Objective To establish a method for the determination of 6 markers in neonatal phenylketonuria（PKU），congenital adrenal hyperplasia（CAH） and congenital hypothyroidism（CH） by ultra-performance liquid chromatography tandem mass spectrometry（UPLC-MS/MS）. Methods The processing conditions of dried blood spots were optimized to establish a UPLC-MS/MS that can simultaneously determine 17-hydroxyprogesterone（17-OHP），androstenedione（ASD），hydrocortisone（COR），phenylalanine（PHE），tyrosine（TYR） and thyroxine（T₄） and carry out methodological validation（linear range，accuracy，sensitivity，precision，reference material determination）. Totally，1 000 normal neonatal dried blood samples，30 CH dried blood samples，30 PKU dried blood samples and 30 CAH dried blood samples for clinical validation were collected，and a reference interval for 6 analytes was preliminarily established. Results The linear ranges for determining PHE，TYR，17-OHP，ASD，COR and T₄ by UPLC-MS/MS were 12.1-980.7 μmol/L，3.7-298.0 μmol/L，6.1-490.2 nmol/L，0.7-56.6 nmol/L，13.8-1 117.3 nmol/L and 32.2-2 606.6 nmol/L，respectively. The minimum determination limits were 0.5 μmol/L，0.7 μmol/L，1.6 nmol/L，0.2 nmol/L，2.6 nmol/L and 3.2 nmol/L，respectively. The intraday precision [coefficient of variation（CV）] of the quality control samples at 3 concentration levels was 2.2%-7.5%，the interday precision（CV） was 2.3%-7.3%，and the spiked recovery rate was 92.5%-115.4%. The relative deviations between the determined values and standard values of the 5 concentrations of the International Society for Neonatal Screening（ISNS） PHE and 17-OHP reference materials were 1.78%-7.01% and 0.70%-10.78%，respectively. Compared to normal newborns，the 17-OHP and（17-OHP+ASD）/COR ratio in CAH newborns were increased（P<0.05）. The PHE level and PHE/TYR ratio of PKU newborns were increased（P<0.05）. The T₄ level in CH newborns was decreased（P<0.05）. The positive rates of 17-OHP and 17-OHP combined with（17-OHP+ASD）/COR screening for CAH were 100% and 100%，respectively. The positive rates of PHE and PHE combined with PHE/TYR screening for PKU were 100% and 100%，respectively. The positive rate of T₄ screening for CH was 40%. Conclusions The UPLC-MS/MS has been established that can simultaneously determine 6 analytes in PKU，CAH and CH. This method has high specificity，speed and sensitivity，and it shortens sample analysis time. The method is expected to provide a new possible option for screening neonatal diseases.

Plasma metabolic markers in patients with insomnia disorder

XU Weixin, CHEN Qing, DU Wenyong, GAO Yan

2023, 38(5):  454-459.  DOI: 10.3969/j.issn.1673-8640.2023.05.009

Asbtract ( 103 )   HTML ( 11)   PDF (1312KB) ( 75 )  

Figures and Tables | References | Related Articles | Metrics

Objective To screen metabolic markers in the blood of patients with insomnia disorder， and to analyze the abnormal metabolic pathways. Methods Totally， 30 patients with insomnia disorder（insomnia disorder group） and 30 healthy subjects（healthy control group） from January to December 2021 at Jiading District Central Hospital of Shanghai University of Medicine and Health Sciences were enrolled. The general data were collected. Ultra performance liquid chromatography tandem mass spectrometry（UPLC-MS/MS） was used to determine plasma metabolites. Based on VIP value， fold change（FC） and P value， significantly differential metabolites were screened between insomnia disorder group and healthy control group. Kyoto Encyclopedia of Genes and Genomes（KEGG） enrichment pathway analysis was performed on differential metabolites. Results Totally， 1 656 metabolites were determined， and 181 differential metabolites between insomnia disorder group and healthy control group were selected. Among them， 115 metabolites were screened for positive ion mode（61 cases of up-regulation and 54 cases of down-regulation）， and the top 10 metabolites with VIP values were ethylenediaminetetraacetic acid， carnitine， N-（1-amino-3， 3-dimethyl-1-oxobutan-2-yl）-1-pentyl-1h-indole-3-formamide， [1-（cyclohexylmethyl）-1h-indole-3-carbonyl]-l-valine methyl ester， di tetracaine， glycerophosphate choline， hypoxanthine， palmitoyl serotonin， deferoxamine d2 and linolenic carnitine. The 66 metabolites（38 cases of up-regulation and 28 cases of down-regulation） were screened for negative ion mode， and the top 10 metabolites with VIP values were citrate， taurine， rhamnocitrin， uric acid， pyruvate， allantoin， 3'， 5'-cyclic inosine monophosphate， D-lactic acid， （+） - 6-aminopenicillanic acid and oleic acid. The enrichment analysis of KEGG pathway showed that the differential metabolites were involved in the central carbon metabolism of cancer， ATP-binding cassette transporter and protein digestion and absorption. Conclusions Screening out representative metabolites and metabolic pathways in patients with insomnia disorder can provide potential targets for the diagnosis and treatment of insomnia disorder.

Relationship between macrophage migration inhibitory factor and carotid vascular calcification in elderly patients with type 2 diabetes mellitus

LIANG Chaohui, ZHAO Fan, LIU Qianwen

2023, 38(5):  460-465.  DOI: 10.3969/j.issn.1673-8640.2023.05.010

Asbtract ( 48 )   HTML ( 3)   PDF (948KB) ( 13 )  

Figures and Tables | References | Related Articles | Metrics

Objective To investigate the relationship between macrophage migration inhibitory factor（MIF）and carotid vascular calcification in elderly patients with type 2 diabetes mellitus（T2DM）. Methods Totally，164 patients with T2DM from Jiangmen People's Hospital from April 2018 to August 2019 were enrolled. According to the vascular calcification of carotid artery，they were classified into non-calcification group，hypocalcification group and hypercalcification group. According to the expressions of MIF，they were classified into high MIF expression group and low MIF expression group. Enzyme-linked immunosorbent assay was used to determine the level of serum MIF. All the patients were followed up for 12 months to understand the incidence of major adverse cardiovascular events. Multivariate Logistic regression analysis was used to evaluate the risk factors of carotid vascular calcification in patients with T2DM. Pearson correlation analysis was used to evaluate the correlation among the indicators. Receiver operating characteristic（ROC） curve was used to evaluate the efficacy of serum MIF in judging carotid vascular calcification in patients with T2DM. Kaplan-Meier survival curve was used to analyze the incidence of major adverse cardiovascular events in elder T2DM patients. Cox proportional hazard regression analysis were used to evaluate the influencing factors of major adverse cardiovascular events in patients with T2DM. Results The course of T2DM，systolic blood pressure，total cholesterol（TC），triglyceride（TG），low-density lipoprotein cholesterol（LDL-C），fasting blood glucose，high-sensitivity C-reactive protein（hs-CRP） and MIF levels in hypercalcification group and hypocalcification group were higher than those in non-calcification group（P<0.05）. The course of T2DM，TC，LDL-C，hs-CRP and MIF levels in hypercalcification group were higher than those in hypocalcification group（P<0.05）. There was no statistical significance in sex，age，body mass index（BMI），proportion of smoking history，diastolic blood pressure，creatinine，urea，aspartate aminotransferase（AST） and alanine aminotransferase（ALT） among the 3 groups（P>0.05）. The results of multivariate Logistic regression analysis showed that the course of T2DM，TC，LDL-C，hs-CRP and MIF were risk factors of carotid vascular calcification in elderly patients with T2DM [odds ratios（OR） were 3.473，2.077，2.230，2.581 and 3.171，95% confidence intervals（CI） were 1.940-6.216，1.179-3.660，1.074-4.632，1.419-4.692 and 1.425-7.055，respectively，P<0.05]. The results of Pearson correlation analysis showed that MIF in elderly patients with T2DM was positively correlated with the course of T2DM，TC，LDL-C and hs-CRP（r=0.389，0.271，0.282 and 0.486，P<0.05）. ROC curve analysis showed that the area under curve（AUC） of MIF in judging carotid vascular calcification in elderly patients with T2DM was 0.881. The incidence of major adverse cardiovascular events in high MIF expression group was higher than that in low MIF expression group（χ²=4.261，P=0.039）. The results of Cox proportional hazard regression analysis showed that hs-CRP，LDL-C and MIF were independent risk factors for major adverse cardiovascular events in elderly T2DM patients with carotid vascular calcification [hazard ratios（HR） were 3.091，3.597 and 3.311，95% CI were 1.628-3.862，3.174-4.928 and 2.497-3.653，respectively，P<0.001]. Conclusions The level of MIF is related to carotid vascular calcification in elderly patients with T2DM，and the high expression of MIF may affect the prognosis of patients by promoting carotid vascular calcification．

Change and significance of NKG2C and granzyme B labeled NK cell subsets in peripheral blood of children with purulent meningitis

YANG Ze, XIE Lili, TIAN Jing, XU Tongtong, ZHOU Guoqing, LIU Baojun, WU Huihui

2023, 38(5):  466-474.  DOI: 10.3969/j.issn.1673-8640.2023.05.011

Asbtract ( 53 )   HTML ( 3)   PDF (1464KB) ( 13 )  

Figures and Tables | References | Related Articles | Metrics

Objective To investigate the change characteristics and clinical significance of natural killer cell receptor 2C（NKG2C） and granzyme B（GZMB） labeled natural killer（NK） cell subsets in peripheral blood of children with purulent meningitis（PM）. Methods Totally，52 children with PM and 52 control children with non-central infectious diseases were enrolled. The children with PM were classified into good prognosis group（44 cases） and poor prognosis group（8 cases） according to the Glasgow Outcome Scale on discharge. NK cell subsets [NKG2C^- gamma-interferon gamma（INF-γ）⁺，NKG2C⁺INF-γ⁺，NKG2C⁺INF-γ^-，NKG2C^-GZMB⁺，NKG2C⁺GZMB⁺，NKG2C⁺GZMB^-] were determined by flow cytometry. The results of cerebrospinal fluid white blood cell（WBC） count，total protein（TP），glucose（Glu） and chlorine（Cl^-），peripheral blood WBC count，the percentage of neutrophil（NEUT%），serum procalcitonin（PCT） and high-sensitivity C-reactive protein（hs-CRP） were collected as well. Spearman rank correlation test was used to analyze the correlations. Receiver operating characteristic（ROC）curves were established to evaluate the values of the indicators in the diagnosis and prognosis of PM. Results The levels of NKG2C⁺，NKG2C^-GZMB⁺，NKG2C⁺GZMB⁺ and NKG2C⁺GZMB^- in PM group were higher than those in control group（P<0.05），and total NK cell percentage was lower than that in control group（P<0.05）. There was no NKG2C^-INF-γ⁺ and NKG2C⁺INF-γ⁺ NK cell in the 2 groups. The percentage of NK2G^-GZMB⁺ NK cells was positively correlated with cerebrospinal fluid WBC count，TP and serum PCT（r values were 0.286，0.558 and 0.740，P<0.05），and it was negatively correlated with cerebrospinal fluid Glu（r=-0.389，P<0.05）. The percentage of NKG2C⁺GZMB⁺ NK cells was positively correlated with cerebrospinal fluid WBC count，TP，peripheral blood WBC count and serum PCT（r=0.509，0.539，0.549 and 0.577，P<0.05），and it was negatively correlated with cerebrospinal fluid Glu（r=-0.391，P<0.05）. NKG2C⁺ GZMB^- NK cell percentage was positively correlated to cerebrospinal Glu level（r=0.624，P<0.05），and it was negatively correlated to cerebrospinal TP（r=-0.328，P<0.05）. The percentage and absolute value of total NK cells in PM group after treatment were higher than those before treatment（P<0.05）. Compared with before treatment，the percentage of NKG2C^-GZMB⁺ and NKG2C⁺GZMB⁺ NK cells in good prognosis group were decreased after treatment（P<0.05），while the percentage of NKG2C⁺GZMB^- NK cells was increased（P<0.05）. The percentage of NKG2C⁺GZMB⁺NK cells in poor prognosis group after treatment was increased（P<0.05）. ROC curve analysis results showed that NKG2C^-GZMB⁺ NK cell percentage had the highest diagnostic efficiency for PM，with the area under curve（AUC） of 0.980，the optimal cut-off value of 8.93%，the sensitivity of 92.31%，and the specificity of 98.08%. After treatment，NKG2C⁺GZMB^- NK cells had the highest efficiency in predicting poor prognosis in children with PM，with the AUC of 0.989，the optimal cut-off value of 5.51%，the sensitivity of 100.00%，and the specificity of 97.73%. Conclusions NKG2C^-GZMB⁺ NK cell subsets have a good value for the early diagnosis in children with PM. NKG2C⁺GZMB^- NK cells have a certain clinical value for prognostic prediction of PM.

Role of coagulation-related monitoring indicators in the course of rivaroxaban anticoagulation for acute venous thromboembolism in elderly patients

XIA Yanyan, XIA Yongquan, SONG Guanghao, XIA Mao

2023, 38(5):  475-478.  DOI: 10.3969/j.issn.1673-8640.2023.05.012

Asbtract ( 68 )   HTML ( 3)   PDF (880KB) ( 82 )  

Figures and Tables | References | Related Articles | Metrics

Objective To study the changes of coagulation-related monitoring indicators in elderly patients with acute venous thromboembolism（VTE） after taking rivaroxaban，and to investigate the appropriate indicators for monitoring the anticoagulation strength and safety of rivaroxaban. Methods Totally，50 elderly patients with acute venous thromboembolism，including 21 males and 29 females，aged（84.25±3.28） years，from Drum Tower Hospital of Nanjing University Medical School from April 2019 to December 2021，were enrolled. Rivaroxaban（10 mg） was administered orally to patients，and the changes on anti-factor Ⅹa activity，coagulation six indicators and coagulation factor activity for VTE patients were observed before and after dosing. Bleeding risk was predicted with related indicators using receiver operating characteristic（ROC） curve. Results Compared with those before taking rivaroxaban，anti-factor Ⅹa activity was higher（P<0.01），prothrombin time（PT），international normalized ratio（INR） and activated partial thromboplastin time（APTT） were higher（P<0.01），and the levels of coagulation factors（Ⅱ，Ⅴ，Ⅶ，Ⅷ，Ⅸ，Ⅹ，Ⅺ and Ⅻ） showed a decreasing trend（P<0.01）. ROC curve analysis showed that PT was a moderate predictor of bleeding risk，and the area under curve was 0.81 [95% confidence interval（CI） 0.66-0.95]. Conclusions PT can be used not only as a preliminary indicator for anticoagulation monitoring with rivaroxaban，but also it has high predictive role for bleeding risk.

Effects of different anticoagulants on phagocytosis，activation and apoptosis of in vitro neutrophil function

CHEN Yi, WANG Jia, XU Zhiwei, ZHAI Yaping, XUAN Weixia

2023, 38(5):  479-483.  DOI: 10.3969/j.issn.1673-8640.2023.05.013

Asbtract ( 65 )   HTML ( 5)   PDF (1248KB) ( 33 )  

Figures and Tables | References | Related Articles | Metrics

Objective To evaluate the effects of different anticoagulants on phagocytosis，activation and apoptosis of in vitro neutrophil function. Methods Totally，30 healthy subjects who underwent physical examination in Henan Provincial People's Hospital from January 2020 to June 2020 were enrolled，and they were classified into 3 groups randomly（10 cases for each group）. The 3 mL peripheral blood was collected with heparin sodium anticoagulant tube（heparin sodium group），2 mL peripheral blood was collected with ethylenediaminetetraacetic acid（EDTA）-K₂ anticoagulant tube（EDTA-K₂ group），and 2 mL peripheral blood was collected with sodium citrate anticoagulant tube（sodium citrate group）. The average fluorescence intensity of OVA-FITC was determined by flow cytometry to determine the phagocytosis ability of neutrophils. The expressions of neutrophil surface activation markers，CD62L and CD11B，were determined to assess the activation of neutrophils. 7AAD and Annexin-V were added to determine the apoptosis of neutrophils. Results The average fluorescence intensity of OVA-FITC phagocytosis neutrophils in EDTA-K₂ group was lower than those in heparin sodium and sodium citrate groups（P<0.05）. CD11B neutrophils in EDTA-K₂ group were lower than those in heparin sodium and sodium citrate groups（P<0.01，P<0.05）. Compared with heparin sodium group，neutrophil apoptosis was increased in EDTA-K₂ and sodium citrate groups（P<0.01，P<0.05）. Conclusions Different anticoagulants can affect the phagocytosis，activation and apoptosis of neutrophils in vitro，but each anticoagulant has different effects. Therefore，appropriate anticoagulant must be selected to accurately determine the phagocytosis，activation and apoptosis of neutrophils.

Effect of peripheral blood neutrophils on interferon-gamma release assay of Mycobacterium tuberculosis

YANG Liyuan, DAI Wanqin, WANG Xiaotong, LOU Xiaoli, WANG Yue, HOU Yanqiang

2023, 38(5):  484-488.  DOI: 10.3969/j.issn.1673-8640.2023.05.014

Asbtract ( 72 )   HTML ( 6)   PDF (1280KB) ( 105 )  

Figures and Tables | References | Related Articles | Metrics

Objective To investigate the effect of peripheral neutrophils on the positive degree of interferon-gamma release assay（IGRA）. Methods From October 2020 to March 2022，the whole blood of suspected tuberculosis patients were collected，according to ages they are classified into teenage（<40 years old），middle age（40-<60 years old） and elderly（≥60 years old） groups. Routine blood examination was performed. Peripheral blood mononuclear cells from tuberculosis patients were isolated for IGRA. The related data and determination results were collected. The results with spot number of test holes-blank control holes 11-14（close to cut-off value） were defined as weak positive，the results with more than 14 were defined as positive，and the results with less than 11 spots were defined as negative. The correlation between neutrophil count and positive degree of IGRA in patients of different ages and sex was analyzed. Two clinical cases were enrolled for verification. Results Among the 234 patients，neutrophil count in weakly positive group [（6.15±3.22）×10⁹/L] was higher than that in positive group [（5.05±2.66）×10⁹/L，P=0.030 2]. The neutrophil number in teenage weakly positive group [（7.54±4.46）×10⁹/L] was higher than that in positive group [（4.00±1.49）×10⁹/L，P=0.000 2]. The number of neutrophils in male patients with weakly positive group [（6.49±3.49）×10⁹/L] was higher than that in positive group [（5.28±2.54）×10⁹/L，P=0.040 8]. Clinical case verification results showed that patients with low neutrophil count were more likely to have positive IGRA results. Conclusions Neutrophil count affects the positive degree of IGRA，so the effect of neutrophils should be excluded in the auxiliary diagnosis of tuberculosis.

Establishment of traceable steroid hormone standard curve for dried blood spot samples

WU Jie, LI Zenghe, CHEN Baorong, KONG Xujing

2023, 38(5):  489-493.  DOI: 10.3969/j.issn.1673-8640.2023.05.015

Asbtract ( 94 )   HTML ( 3)   PDF (1167KB) ( 65 )  

Figures and Tables | References | Related Articles | Metrics

Objective To establish a traceable standard curve for 19 steroid hormones in dried blood spot samples by liquid chromatography tandem mass spectrometry（LC-MS/MS）. Methods Ethylenediaminetetraacetic acid fresh anticoagulated whole blood was collected，blood cells were washed with 0.9 NaCl saline，and the washed blood cells and steroid hormone-free plasma were mixed in the ratio of 0.55∶0.45. Simulated whole blood samples was prepared. Different amounts of steroid hormones were added to simulated whole blood samples according to the prepared standard curve concentration in order to prepare different concentrations of steroid hormone standard curve dried blood spot samples. Steroid hormone concentrations in standard curve dried blood spot samples were determined by LC-MS/MS，and its linear range was analyzed. The performance of dried blood spot steroid hormone standard curve was verified. Results The linear ranges of dried blood spot steroid hormone standard curve for pregnenolone，progesterone，cortisone and dehydroepiandrosterone were 0.25-100 ng/mL. 11-Deoxycorticosterone，21-deoxycortisol，18-hydroxycortisol and 11-keto testosterone were in the linear range of 0.05-20 ng/mL. Corticosterone and 11-hydroxytestosterone were in the linear range of 0.025-10 ng/mL. 17-Hydroxyprogesterone，18-hydroxycorticosterone，11-deoxycortisol，androstenedione，11-hydroxyandrostenedione and testosterone were in the linear range of 0.1-40 ng/mL. Cortisol was in the linear range of 1.5-600 ng/mL，17-hydroxypregnenolone was in the linear range of 0.2-80 ng/mL，and dehydroepiandrosterone sulfate was in the linear range of 25-10 000 ng/mL with a correlation coefficient of squared（r²）≥0.997. Conclusions The established steroid hormone standard curves for dried blood spot samples have good linearities and can achieve the traceability of steroid hormone determination results in dried blood spots samples.