Table of Content

30 April 2024, Volume 39 Issue 4

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Clinical value and prospect of peripheral blood immune indexes in cancer patients

ZHANG Han, GUO Lin, LU Renquan, CHEN Fuxiang

2024, 39(4):  311-316.  DOI: 10.3969/j.issn.1673-8640.2024.04.001

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Immune microenvironment formed by local immune reaction and inflammatory responses plays a role in tumorigenesis，development and metastasis. It consists of tumor cells，immune cells（neutrophils，macrophages and lymphocytes） and the cytokines and chemokines they produce. These components interact with each other to form a complex network that affects the body's immune status. Peripheral blood immune indexes can be a valuable reference for the auxiliary diagnosis，prognosis and efficacy prediction of cancer patients，particularly in evaluating immunotherapy，which has a potential for clinical application. However，interpreting test results can be challenging due to the significant differences in individual immune status among cancer patients. This review focuses on the clinical value and potential applications of peripheral blood immune indexes in cancer patients.

Research progress on the regulation of regulatory T cells in ovarian cancer tumor microenvironment

ZHANG Heng, ZHENG Hui, LU Renquan

2024, 39(4):  317-323.  DOI: 10.3969/j.issn.1673-8640.2024.04.002

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Ovarian cancer is one of the main causes of female cancer death. Although immunotherapy has been used clinically for ovarian cancer，the effective rate of immunotherapy in ovarian cancer is low. There are immunosuppressive factors in tumor microenvironment，the progression of ovarian cancer may be associated with imbalance of immune system. As an important suppressive T cell subset，regulatory T cells（Treg） play roles in tumor immunity，autoimmunity and maintaining immune tolerance. Treg plays an important role in tumor microenvironment of ovarian cancer，participating in anti-tumor immune tolerance and associating with the progression and prognosis of ovarian cancer. This review mainly focuses on the research progress of tumor microenvironment on Treg in ovarian cancer.

Predictive role of neutrophil-to-lymphocyte ratio for distant metastasis in solid tumor patients

LI Yue, TENG Xiaoyan, DING Siying, HU Liuping, DU Yuzhen

2024, 39(4):  324-329.  DOI: 10.3969/j.issn.1673-8640.2024.04.003

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Objective To investigate the predictive value of neutrophil-to-lymphocyte ratio（NLR） in distant metastasis of various solid tumors. Methods A total of 259 patients diagnosed with solid tumors at the Sixth Affiliated People's Hospital of Shanghai Jiao Tong University were enrolled，which included 94 patients with digestive system tumors，78 patients with respiratory system tumors，37 patients with breast tumors and 50 patients with other types of solid tumors. There were 104 cases in the group without distant metastasis（TNM stage Ⅰ，Ⅱ，Ⅲ） and 155 cases in the group with distant metastasis（TNM stage Ⅳ）. The blood routine indicators，lymphocyte subsets and natural killer（NK） cells were determined，and the NLR，platelet-to-lymphocyte ratio（PLR） and lymphocyte-to-monocyte ratio（LMR）were calculated. Binary Logistic regression analysis was used to evaluate the risk factors of distant metastasis in patients with solid tumors. Multiple Logistic regression analysis was used for stratified analysis. Receiver operating characteristic（ROC） curve was used to evaluate the efficacy of each indicator in identifying distant metastasis in patients with solid tumors. Totally，47 patients with solid tumors from January to August 2021 were followed up for 36 months，with the occurrence of distant metastasis as the end event. Disease-free survival（DFS） of solid tumor patients was analyzed by Kaplan-Meier survival curve. Results There was statistical significance on male proportion，white blood cell （WBC） count，red blood cell（RBC） count，hemoglobin（Hb），hematocrit（HCT），the absolute value of neutrophils（NEUT#），the absolute value of lymphocytes（LYMPH#），the absolute value of monocytes （MO#），NLR，PLR and LMR between distant metastasis group and non-distant metastasis group（P<0.05）. There was no statistical significance in the other indicators（P>0.05）. Multivariate Logistic regression analysis showed that NLR was an independent risk factor for distant metastasis in solid tumor patients [odds ratio（OR）=1.488，95% confidence interval（CI） 1.250-1.771，P<0.001]. ROC curve analysis results showed that the area under curve of distant metastasis in solid tumor patients determined by NLR was 0.774. According to the optimal cut-off value of NLR（2.34），solid tumor patients were classified into high NLR（NLR>2.34） group and low NLR（NLR≤2.34） group. C+++ompared with low NLR group，T helper cells，cytotoxic T cells，NK cells and double-negative T cells were decreased in high NLR group（P<0.05），while B cells were increased（P=0.001），and there was no statistical significance in double-positive T cells（P=0.676）. NLR was a risk factor for distant metastasis in different types of solid tumors（P<0.05）. The NLR of TNM stage Ⅳ patients was higher than those of TNM stage Ⅰ，Ⅱ and Ⅲ patients（P<0.001），and there was no statistical significance among TNM stage Ⅰ，Ⅱ and Ⅲ patients（P>0.05）. The DFS in high NLR group was shorter than that in low NLR group（P<0.001），and the incidence of distant metastasis in high NLR group was higher than that in low NLR group（P<0.001）. Conclusions NLR can predict the distant metastasis of different types of solid tumors. The increase of NLR may be related to the enhancement of immunosuppression in patients with solid tumors.

Relationship between peripheral blood T lymphocyte subsets and tumor progression in patients with colorectal cancer

ZHENG Hui, CHEN Yingxiu, YE Lüyin, LU Renquan, GUO Lin

2024, 39(4):  330-335.  DOI: 10.3969/j.issn.1673-8640.2024.04.004

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Objective To investigate the relationship between peripheral blood T lymphocyte subsets and tumor progression in patients with colorectal cancer. Methods Totally，100 newly diagnosed colorectal cancer patients in Shanghai Cancer Center of Fudan University from January to December 2023 were enrolled as colorectal cancer group. All the patients were classified into stage Ⅰ（8 cases），stage Ⅱ（47 cases），stage Ⅲ（34 cases） and stage Ⅳ（11 cases） according to TNM stages. There were 55 cases with lymphatic metastasis and 45 cases without lymphatic metastasis. Totally，80 healthy subjects were enrolled as healthy control group. Flow cytometry was used to determine the percentages of CD3⁺T cells（CD3⁺%），CD3⁺CD4⁺T cells（CD3⁺CD4⁺%），CD3⁺CD8⁺T cells（CD3⁺CD8⁺%），CD3⁺CD4^-CD8^-T cells [double-negative T cells （DNT）]（DNT%），CD4⁺CD25⁺CD127^low regulatory T cells（CD4⁺Treg%），CD3⁺PD-1⁺T cells（CD3⁺PD-1⁺%），and the CD4⁺/CD8⁺ ratio was calculated. The tumor biomarkers，carcinoembryonic antigen（CEA），carbohydrate antigen（CA）19-9，CA72-4，CA50 and CA242，were also determined. The efficacy of various indicators in determining lymphatic metastasis was evaluated by receiver operating characteristic（ROC） curve. Results Compared with healthy control group，the CD3⁺CD8⁺% in colorectal cancer group was increased（P<0.05），while the CD3⁺CD4⁺%，DNT% and CD4⁺/CD8⁺ ratio were decreased（P<0.05）. There was no statistical significance in CD3⁺% between the 2 groups（P>0.05）. There was statistical significance in CD4⁺/CD8⁺ ratio，CD4⁺Treg% and CD3⁺PD-1⁺% among colorectal cancer patients with different TNM stages（P<0.05）. Compared with colorectal cancer patients without lymphatic metastasis，colorectal cancer patients with lymphatic metastasis showed a decrease in CD4⁺/CD8⁺ ratio（P<0.001），while CD4⁺Treg% and CD3⁺PD-1⁺% were increased（P<0.001）. The area under curve（AUC） for single and combined determinations of CD4⁺/CD8⁺ ratio，CD4⁺Treg% and CD3⁺PD-1⁺% to determine lymphatic metastasis in colorectal cancer patients were 0.762，0.755，0.765 and 0.825，respectively. If taking any one of CEA，CA19-9，CA72-4，CA50 or CA242 positive as tumor biomarker positive，the patients for TNM stage Ⅰ were all negative，while the positive rates for stage Ⅱ，Ⅲ and Ⅳ were 48.9%（23/47），52.9%（18/34） and 90.9%（10/11），respectively. In patients of TNM stage Ⅱ and Ⅲ，the CD4⁺/CD8⁺ ratio in tumor biomarker positive patients was lower than that in negative patients（P<0.001），and CD4⁺Treg% was higher than that in negative patients（P<0.001）. There was no statistical significance in CD3⁺PD-1⁺%（P>0.05）. Conclusions The patients with colorectal cancer have the enhancement of immunosuppressive function，and T lymphocyte subsets have relations with tumor progression and tumor biomarker expression.

Roles of serum cytokine determinations in progression of epithelial ovarian cancer

YAN Tianqing, YANG Xiaoxiang, GUO Lin, LU Renquan

2024, 39(4):  336-342.  DOI: 10.3969/j.issn.1673-8640.2024.04.005

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Objective To investigate the clinical roles of serum 12 cytokines [interleukin（IL）-1β，IL-2，IL-4，IL-5，IL-6，IL-8，IL-10，IL-12，IL-17，interferon-gamma（INF-γ），interferon-alpha（IFN-α），tumor necrosis factor-alpha（TNF-α）] determinations in patients with epithelial ovarian cancer. Methods From May 2021 to May 2023，67 patients with epithelial ovarian cancer（ovarian cancer group），22 patients with benign ovarian tumors（ovarian cysts，cystadenomas and so on）（benign group） and 21 healthy subjects（healthy control group） were enrolled from Shanghai Cancer Center of Fudan University. The clinical data of all the subjects were collected，and serum levels of 12 cytokines were determined. All the patients were followed up until September 1st，2023. The survival of all the patients was analyzed by Kaplan-Meier curve. Results Compared with healthy control group，serum levels of IL-5，IL-8，IL-10，IL-17，TNF-α and IFN-γ were decreased in ovarian cancer group（P<0.05），and the level of TNF-α in ovarian cancer group was lower than that in benign group（P<0.05）. There was no statistical significance in the other cytokines among the 3 groups（P>0.05）. The positive rate of IL-6 was the highest in ovarian cancer group（85.1%），followed by IL-10（49.3%） and IL-8（40.3%），and the positive rates of the other cytokines were <40%. The negative rates of IL-4，TNF-α and IFN-γ were higher than positive rates in patients with carbohydrate antigen（CA）125 and human epididymis protein 4（HE4）-positive epithelial ovarian cancer（P<0.05）. The positive rates of serum T helper cell（Th）2 cytokines（IL-4，IL-6，IL-10） were higher than those of Th1 cytokines（IL-2，TNF-α，IFN-γ） in patients with advanced the International Federation of Gynecology and Obstetrics（FIGO） stage，serous histological type，ascites and lymphatic metastasis（P<0.05）. There was statistical significance in the positive rate of Th1 cytokines among different FIGO stages（P=0.001）. However，no statistical significance was found in the positive rates of Th1 and Th2 cytokines for epithelial ovarian cancer patients with different clinical characteristics（P>0.05）. The progression-free survival（PFS） of the positive group of IL-2，IL-6 and IL-10 was shorter than that of the corresponding negative group（Log-rank χ² values were 3.567，4.463 and 3.997，P=0.046，0.013 and 0.035，respectively）. The PFS in the positive group of IL-17 was longer than that in the corresponding negative group（Log-rank χ² value was 3.659，P=0.043），but there was no statistical significance in PFS between the positive group and the negative group of the other cytokines（P>0.05）. Conclusions Cytokine determination is helpful to monitor inflammatory status and immune function in patients with epithelial ovarian cancer，and has a certain value in judging disease progression and evaluating prognosis.

Effect of highly expressed IL-35 on Th1 and Th17 plasticity in colorectal cancer

XU Yunchuan, HUANGFU Yuchan, MA Yanhui

2024, 39(4):  343-350.  DOI: 10.3969/j.issn.1673-8640.2024.04.006

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Objective To investigate the effect of highly expressed inhibitory cytokine interleukin（IL）-35 on the plasticity of T helper cell 1（Th1） and T helper cell 17（Th17）in colorectal cancer tissues. Methods A total of 90 patients with colorectal cancer（colorectal cancer group） and 28 healthy subjects（healthy control group） were enrolled from Xinhua Hospital of Shanghai Jiao Tong University School of Medicine from March 2019 to February 2020. Flow cytometry was used to determine Th1 percentage（Th1%） and Th17 percentage（Th17%） in peripheral blood of colorectal cancer group and healthy control group. The effects of highly expressed interferon-gamma（IFN-γ） and IL-17A on overall survival and poor prognosis of colorectal cancer were analyzed based on Kaplan-Meier Plotter database. Immunohistochemistry was used to determine the expression of IL-35 in cancer tissues and adjacent tissues（2 cm from tumor margin） of colorectal cancer patients. Serum IL-35 levels in colorectal cancer and healthy control groups were determined by enzyme-linked immunosorbent assay. The effect of IL-35 on the secretion of IL-17A and IFN-γ by Th1 was observed based on the induced Th1 differentiation induction experiment in vitro. Results Compared with healthy control group，Th1% in colorectal cancer group was decreased（P=0.019），and Th17% was increased（P<0.001）. The r² values of Th1% and Th17% in colorectal cancer group（0.393 4） were different from those in healthy control group（0.041 0）（P=0.007）. Low expression of IFN-γ mRNA was a risk factor for shorter survival in colorectal cancer patients [hazard ratio（HR）=0.74，95% confidence interval（CI）0.58-0.94，P=0.013]. High expression of IL-17A mRNA was a risk factor for shorter survival in colorectal cancer patients（HR=1.26，95%CI 1.02-1.59，P=0.020）. The expression of IL-35 in colorectal cancer tissues was higher than that in adjacent tissues（P<0.001）. Serum IL-35 levels in patients with stage Ⅰ to Ⅱ（early stage） and stage Ⅲ to Ⅳ（advanced stage） colorectal cancer were higher than those in healthy control group（P<0.05）. According to the results of Th1 differentiation induction experiment in vitro，IL-35 can decrease the expression of IFN-γ and increase the expression of IL-17A in Th1. Conclusions The proportions of Th1 and Th17 in peripheral blood of patients with colorectal cancer show abnormal changes，and the expression of IL-35 increases. High expression of IL-35 can decrease the expression of Th1 IFN-γ and increase the expression of IL-17A.

Apolipoprotein A1 antibodies reduce sperm motility by inhibiting ionophore Ca²⁺ A23187-induced Ca²⁺ influx

LIN Jing, CHI Xiuping, LIANG Zhenlong, XIANG Daijun, WANG Chengbin

2024, 39(4):  351-357.  DOI: 10.3969/j.issn.1673-8640.2024.04.007

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Objective To analyze the influence of apolipoprotein A1（apo A1） antibodies on ionophore Ca²⁺ A23187-induced Ca²⁺ influx，and to investigate the relation of apo A1 antibodies with sperm motility. Methods Totally，38 healthy male semen samples were collected. Immunofluorescence was used to observe the localization of apo A1 in human spermatozoa and the effect of apo A1 antibody on sperm acrosome reaction. Western blot was used to determine the tyrosine phosphorylation of proteins related to sperm capacitation. Samples were treated with normal rabbit IgG（normal rabbit IgG group） and 40 μg·mL^-1 apo A1 antibody（apo A1 antibody group）. Normal semen samples were used as blank control group，and the difference of total sperm motility and the percentage of progressively motile sperm were compared. Fluo-4AM-loaded semen samples induced by 10 μmol·L^-1 A23187 for 30 min were treated with normal rabbit IgG and 10，20 and 40 μg·mL^-1 apo A1 antibodies，respectively. Fluo-4AM-loaded sperm in blank control group were incubated，and Ca²⁺ levels in sperm were determined by flow cytometry. Results The apo A1 protein is localized in the head of human sperm. The total sperm motility and the percentage of progressively motile sperm in apo A1 antibody group were lower than those in blank control group and normal rabbit IgG group（P<0.001）. There was no statistical significance between blank control group and normal rabbit IgG group（P>0.05）. Flow cytometry showed that the fluorescence intensity of intracellular Ca²⁺ increased after induction with A23187 and decreased after treatment with apo A1 antibody in a dose-dependent manner（P<0.01）. The acrosome reaction rate of 40 μg·mL^-1 apo A1 antibody+A23187 group was lower than those of A23187 group and normal rabbit IgG+A23187 group（P<0.001），and apo A1 antibody had no effect on the spontaneous acrosome reaction of sperm. The results of western blot showed that the protein tyrosine phosphorylation of sperm before and after apo A1 antibody treatment did not change significantly. Conclusions The apo A1 could decrease sperm motility by decreasing Ca²⁺ influx and inhibit the acrosome reaction induced by ionophore Ca²⁺ A23187.

Correlation of serum ANGPTL4，HSP70 and IL-34 levels and insulin resistance in overweight and obese patients with type 2 diabetes mellitus

LI Fang, LI Zhihong, YAO Mingyan, YIN Fei

2024, 39(4):  358-362.  DOI: 10.3969/j.issn.1673-8640.2024.04.008

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Objective To investigate the correlation between serum angiopoietin-like protein 4（ANGPTL4），heat shock protein（HSP）70，interleukin-34（IL-34） levels and insulin resistance in overweight and obese patients with type 2 diabetes mellitus（T2DM）. Methods From May 2020 to May 2022，182 patients with T2DM who were treated in Baoding First Central Hospital were enrolled as research group. According to the World Health Organization obesity diagnostic criterion，they were classified into overweight and obese group（90 cases） and normal weight group（92 cases）. Totally，90 healthy subjects were enrolled as healthy control group，including 40 overweight and obese subjects（overweight and obese control group） and 50 normal weight subjects（normal weight control group）. Serum ANGPTL4，HSP70，IL-34，insulin and blood glucose levels were determined，and homeostasis model assessment for insulin resistance（HOMA-IR） was calculated. Serum levels of ANGPTL4，HSP70，IL-34 and HOMA-IR in T2DM group were determined again after 3 months of treatment. Pearson correlation analysis was used to evaluate the correlation between serum ANGPTL4，HSP70，IL-34 and HOMA-IR. Results Compared with healthy control group and normal weight T2DM group，serum ANGPTL4 and HSP70 levels in overweight and obese T2DM group were decreased（P<0.05），and serum IL-34 level and HOMA-IR were increased（P<0.05）. Compared with healthy control group，the levels of serum ANGPTL4 and HSP70 in normal weight T2DM group were decreased（P<0.05），and the levels of serum IL-34 and HOMA-IR were increased（P<0.05）. Serum ANGPTL4 and HSP70 were negatively correlated with HOMA-IR（r values were -0.733 and -0.758，P<0.001），and IL-34 was positively correlated with HOMA-IR（r=0.705，P<0.001）. After treatment，serum ANGPTL4 and HSP70 levels were increased in overweight and obese T2DM group and normal weight T2DM group，while serum IL-34 level and HOMA-IR were decreased. Compared with overweight and obese T2DM group，serum ANGPTL4 and HSP70 in normal weight T2DM group were increased（P<0.05），and serum IL-34 level and HOMA-IR were decreased（P<0.05）. Conclusions ANGPTL4，HSP70 and IL-34 are correlated with insulin resistance in overweight and obese patients with T2DM，which may be used as the indicators for monitoring the efficacy of overweight and obese patients with T2DM.

Relation of C-reactive protein-to-albumin ratio and systemic immune-inflammation index with pathological characteristics and prognosis of serous ovarian cancer patients

ZHOU Yufei, LI Yunhui, LI Bingyi, HOU Qingxia

2024, 39(4):  363-368.  DOI: 10.3969/j.issn.1673-8640.2024.04.009

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Objective To investigate the relation of C-reactive protein（CRP）-to-albumin（Alb） ratio（CAR） and systemic immune-inflammation index（SII） with pathological characteristics and prognosis of serous ovarian cancer patients. Methods A total of 132 patients with serous ovarian cancer were enrolled from Luoyang Central Hospital from January 2017 to January 2021. The general clinical data and preoperative platelet（PLT） count，neutrophil（NEUT） count，lymphocyte（LY） count，CRP，Alb determination results were collected，and CAR and SII were calculated. According to the medians of CAR and SII，the patients were classified into low CAR group，high CAR group and low SII group，high SII group，respectively. The differences of pathological characteristics，recurrence free survival（RFS） and overall survival（OS） among different CAR and SII groups were analyzed. Both univariate and multivariate Cox regression models were used to analyze the influencing factors of RFS and OS in patients with serous ovarian cancer. Results The International Federation of Gynecology and Obstetrics（FIGO） stage，histological grade and lymphatic metastasis in high CAR group and high SII group had statistical significance with those in low CAR group and low SII group，respectively（P<0.05）. The proportions of residual tumor diameter <1 cm in high CAR group and high SII group were lower than those in low CAR group and low SII group，respectively（P<0.05）. The RFS and OS of high CAR group and high SII group were lower than those of low CAR group and low SII group，respectively（P<0.05）. FIGO stage（Ⅲ+Ⅳ），high CAR and high SII were independent risk factors for postoperative RFS in patients with serous ovarian cancer [hazard ratios（HR） were 2.258，2.665 and 4.432，95% confidence intervals（CI） were 1.125-4.534，1.401-5.069 and 2.227-8.821，respectively]. FIGO stage（Ⅲ+Ⅳ），carbohydrate antigen 125（CA125）（≥35 U/L），high CAR and high SII were independent risk factors for postoperative OS in patients with serous ovarian cancer（HR were 4.574，4.417，3.167 and 5.500，95%CI 1.660-12.607，1.426-13.686，1.392-7.206 and 2.254-13.424，respectively）. Conclusions CAR and SII in patients with serous ovarian cancer are related to FIGO stage，histological grade and lymphatic metastasis，and preoperative high CAR and SII are independent risk factors for poor prognosis of patients with serous ovarian cancer，which can be used as reference indexes for the prognosis evaluation of patients with serous ovarian cancer.

Validation of HIV-1 integrase genotyping sequence assay

WANG Xuqin, LIN Qianru, FENG Wanqing, DONG Yuan, YU Xiaolei, LIU Changhe, NING Zhen, SHEN Xin, PAN Qichao, LIN Yi

2024, 39(4):  369-375.  DOI: 10.3969/j.issn.1673-8640.2024.04.010

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Objective To validate laboratory-developed human immunodeficiency virus-1（HIV-1）integrase genotyping sequence assay which can be applied to assess the level of genotyping drug resistance to HIV-1 integrase inhibitors. Methods According to the World Health Organization self-developed gene sequence analysis method validation recommendation，RNA was extracted from 20 HIV-1 positive samples，and the gene fragments of HIV-1 integrase region were amplified and sequenced. The accuracy of laboratory-developed HIV-1 integrase genotyping sequence assay was evaluated by comparison with virus quality assurance（VQA） consensus. Amplification sensitivity was evaluated by amplification success rate. Precision and reproducibility were evaluated by repeated tests of the same sample. Results All the 20 samples had a nucleotide agreement rate of >98% with VQA consensus. The multi-well amplification rate of 10 high viral load samples（>10 000 copies·mL^-1） and 5 low viral load samples （1 000-5 000 copies·mL^-1） was 100%. The results of 5 complex wells of 4 samples of the same batch，as well as 5 assays of 5 samples，met the requirement of nucleotide agreement rate of >98% in 90% sample pairwise comparison. Conclusions The accuracy，amplification sensitivity，precision and reproducibility of this assay meet all the requirements and are suitable for HIV-1 integrase genotyping.

Prognosis of RDW-SD and RDW-CV in breast cancer patients with lymphatic metastasis

HUANG Lei, TANG Wenjia, ZHOU Yan, ZHOU Jiaye, ZHANG Chunyan, YANG Jing, WANG Beili, PAN Baishen, GUO wei

2024, 39(4):  376-381.  DOI: 10.3969/j.issn.1673-8640.2024.04.011

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Objective To investigate the relationship between preoperative red blood cell distribution width-standard deviation（RDW-SD） and red blood cell distribution width-coefficient of variation（RDW-CV） and prognosis in breast cancer patients with lymphatic metastasis. Methods From January 2018 to February 2022，156 female breast cancer patients with lymphatic metastasis confirmed by postoperative pathology in Xiamen Hospital of Zhongshan Hospital of Fudan University were enrolled. The RDW-SD，RDW-CV，clinicopathological indicators and prognostic data of patients were collected. Cox regression model was used to look for risk factors for poor prognosis. The predictive values of RDW-SD and RDW-CV in poor prognosis were analyzed by receiver operating characteristic（ROC） curve. According the optimal values of RDW-CV and RDW-SD，the 156 breast cancer patients with lymphatic metastasis were classified into high and low expression groups. Disease-free survival was calculated using Kaplan-Meier survival analysis. Results There was statistical significance in RDW-SD and RDW-CV in patients with different TNM stages（P<0.001）. RDW-CV in diagnosing Ⅳ stage and evaluating poor prognosis [areas under curves（AUC）=0.838 and 0.773，respectively，P<0.001] were more effective than RDW-SD（AUC=0.780 and 0.729，respectively，P<0.05）. Preoperative RDW-CV was an independent risk factor for poor prognosis [hazard ratio（HR）=1.46，95% confidence interval（CI） 1.13-1.89，P=0.004]. Postoperative disease-free survival rate of RDW-CV high expression group and RDW-SD high expression group were lower than those of low expression groups（P<0.001）. Conclusions RDW-CV is superior to RDW-SD in preoperative prognosis assessment of breast cancer patients with lymphatic metastasis，and high RDW-CV is an independent risk factor for poor prognosis.

Relation of drug resistance，serotypes and genotypes of Group B Streptococcus in perinatal pregnant females and pregnancy outcomes

ZHAO Yanan, XIAO Weili, CAO Qixin, YAN Yan, CUI Xiuge, ZHAO Jianping

2024, 39(4):  382-386.  DOI: 10.3969/j.issn.1673-8640.2024.04.012

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Objective To analyze the correlation of drug resistance，serotypes and genotypes of Group B Streptococcus（GBS） in perinatal pregnant females and pregnancy outcomes，and to provide a reference for clinical drug use and vaccine development. Methods A total of 123 GBS positive perinatal pregnant females（study group） and 123 GBS negative pregnant females（control group） were enrolled from January 2020 to December 2022 in Inner Mongolia People's Hospital. The pregnancy outcome of the 2 groups was compared. The drug susceptibility of GBS was analyzed in vitro. Serotyping and genotyping of GBS were performed by latex agglutination and multi-locus sequence typing（MLST）. The correlation between drug resistance and pathogenicity of GBS in different serotypes and genotypes was analyzed. Results The susceptibility rates of GBS to vancomycin，quinupristin-dafoputin，linezolid，gentamicin，ampicillin，penicillin and ceftriaxone were all >91.00%，and they were resistant to furantoin，levofloxacin，clindamycin and erythromycin in different degrees. The serotypes of GBS were mainly type Ⅲ，Ⅰb and Ⅰa，and the genotypes were mainly ST19，ST17 and ST23. Type Ⅲ GBS genotypes were mainly ST19 and ST17，type Ⅰb was mainly ST10 and ST12，and type Ⅰa was mainly ST23. Type Ⅲ，type Ⅰa and ST19，ST17 GBS had the highest resistance rate to erythromycin，and the difference in resistance rate to furantoin，levofloxacin，clindamycin and erythromycin was statistically significant（P<0.05）. The drug resistance rate of different serotypes of GBS to furantoin，levofloxacin，clindamycin and erythromycin was statistically significant（P<0.05），and the drug resistance rate of type Ⅲ GBS to the 4 antibiotics was the highest. The study group was more likely to have adverse pregnancy outcomes（P<0.05）. The incidence of type Ⅰa and ST23 GBS was higher in pregnant females. Type Ⅲ GBS clones mostly caused invasive infection in neonates，and ST17 was the main type. Conclusions GBS is highly sensitive to penicillins，cephalosporins and vancomycin. The main serotypes of GBS are type Ⅲ，type Ⅰa and type Ⅰb，and the main genotypes are ST19，ST17 and ST23. GBS infection is more likely to cause adverse pregnancy outcomes，type Ⅰa and ST23 GBS are more likely to cause colonization infection in pregnant females，and type Ⅲ and ST17 GBS are more likely to cause invasive infection in neonates.

Clinical role of lymphocyte subsets in graft-versus-host disease patients after hematopoietic stem cell transplantation

LIANG Zhenlong, GUO Yuni, WANG Nan, WANG Jianan, LIU Jiayu, LIU Peipei, XIANG Daijun, WANG Chengbin, LI Mianyang

2024, 39(4):  387-392.  DOI: 10.3969/j.issn.1673-8640.2024.04.013

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Objective By dynamically monitoring the lymphocyte subsets of blood disease patients at 30，60 and 90 d after allogeneic hematopoietic stem cell transplantation（allo-HSCT），to investigate the correlation between lymphocyte subset changes and the occurrence of acute graft-versus-host disease（aGVHD） after allo-HSCT and the clinical significance of lymphocyte subset monitoring for aGVHD. Methods Totally，33 patients who underwent allo-HSCT at the First Medical Center of Chinese People's Liberation Army General Hospital from January 2019 to December 2021 were enrolled. According to whether the patients developed aGVHD，the patients were classified into aGVHD group（19 cases） and non-aGVHD group（14 cases）. Flow cytometry was used to determine lymphocyte subsets（CD3⁺，CD4⁺，CD8⁺，CD19⁺，CD16⁺CD56⁺） and regulatory T cells（CD4⁺CD25⁺FOXP3⁺） at 30，60 and 90 d after transplantation. The changes of lymphocyte subsets were characterized，and the difference in lymphocyte subsets between aGVHD and non-aGVHD groups was compared. Results The relative count of natural killer（NK） cells reached a high level at 30 d after transplantation and gradually decreased to normal levels at 60 d，while B cells remained at a low level for a long time. T cells were at a low level on the 30th day and returned to normal levels after 60 d. The levels of CD8⁺ were higher than those of CD4⁺ at each monitoring time point. For the relative counts of lymphocyte subsets，the levels of NK cells，total T，CD8⁺，CD8⁺CD28⁺ and CD8⁺CD28^- had statistical significance between each time point（P<0.05）. For the absolute counts，there was statistical significance among the 3 time points of CD3⁺，CD4⁺ and CD8⁺ （P<0.05），but there was no statistical significance between NK cells and B cell subsets. There was statistical significance in CD4⁺ on the 60th day between aGVHD and non-aGVHD patients（P<0.05）. There was statistical significance in regulatory T cells between the 2 groups on the 30th day（P<0.05），while the differences in other lymphoid subpopulations had no statistical significance（P>0.05）. Conclusions For patients undergoing hematopoietic stem cell transplantation（HSCT），regular determination of lymphocyte subsets is of importance for monitoring the process of immune reestablishment. Dynamic monitoring of regulatory T cells is helpful for timely diagnosis and prevention of aGVHD，thus further improving the survival rate of patients.

Relationship between serum homocysteine，cystatin C levels and cognitive impairment in patients with herpes simplex encephalitis

YU Jiahui, ZHOU Lin, GUO Gang, ZHOU Jianmei

2024, 39(4):  393-399.  DOI: 10.3969/j.issn.1673-8640.2024.04.014

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Objective To investigate the relationship between serum homocysteine（Hcy），cystatin C（Cys C） levels and cognitive impairment in patients with herpes simplex encephalitis（HSE）. Methods Totally，106 HSE patients in Zhuji Hospital of Traditional Chinese Medicine from April 2018 to May 2022 were enrolled and classified into cognitive normal group and cognitive impairment group according to Montreal cognitive assessment（MoCA） scores. The general data were collected，and blood lipids，fasting blood glucose，serum creatinine（SCr），uric acid（UA），high-sensitivity C-reactive protein（hs-CRP），Hcy，Cys C levels and cerebrospinal fluid pressure，cerebrospinal fluid protein level，cerebrospinal fluid white blood cell count were determined. LASSO regression analysis and 10-time cross-validation method were used to obtain a subset of predictors of HSE patients with cognitive impairment，and multi-variate Logistic regression analysis was used to evaluate the risk factors of cognitive impairment in HSE patients. Smooth curve fitting was used to analyze the relationship between serum Hcy and CysC levels and cognitive impairment. A nomogram prediction model was constructed for evaluating the occurrence of cognitive impairment in HSE patients. The efficiency was evaluated. Results There was statistical significance in age，years of education，the National Institute of Health Stroke Scale（NIHSS） score，hs-CRP，Hcy，MoCA score，CysC and cerebrospinal fluid pressure between cognitive impairment group and cognitive normal group（P<0.05）. Totally，7 predictors（years of education，NIHSS score at admission，Hcy，hs-CRP，MoCA score，Cys C and cerebrospinal fluid pressure） were selected by LASSO regression analysis. Multi-variate Logistic regression analysis showed that hs-CRP，Hcy，CysC and cerebrospinal fluid pressure were independent risk factors for cognitive impairment in HSE patients（P<0.05），and MoCA score was an independent protective factor for cognitive impairment in HSE patients（P<0.05）. The smooth curve fitting results showed that Hcy and CysC were linearly positively correlated with cognitive impairment. Threshold analysis results showed that when Hcy ≥14.35 μmol·L^-1 was increased by 1 μmol·L^-1，the risk of cognitive impairment in HSE patients increased by 20%[odds ratio（OR）=0.80，95% confidence interval（CI） 0.68-0.94，P<0.05]. When Cys C≥1.72 mg·L^-1，the probability of cognitive impairment in HSE patients increased by 8% for every 0.2 mg·L^-1 increase in Cys C level（OR=0.92，95%CI 0.88-0.98，P<0.05）. The nomogram prediction model based on independent influencing factors had high differentiation，accuracy and clinical applicability. Conclusions Serum Hcy and Cys C levels are related to cognitive impairment in HSE patients. The nomogram prediction model based on cerebrospinal fluid pressure，Hcy，Cys C，MoCA score and hs-CRP was helpful to predict the occurrence of cognitive impairment in HSE patients.

Application progress of exosomal glycosylation in diagnosis and treatment of tumors

WANG Rong, GAO Chunfang

2024, 39(4):  404-409.  DOI: 10.3969/j.issn.1673-8640.2024.04.016

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As an important medium of cellular communication，exosomes transport RNA，proteins and lipids between cells. The glycan structure is an important component of exosomes. Glycosylation，as one of the important ways of protein post-translational modification，enriches the function and information meaning of proteins by transferring glycan structure to proteins under the action of glycosyltransferase and provides new biomarkers and intervention targets for the diagnosis and treatment of diseases. There are abnormal N-glycan structures consistent with tumor cells in the exosomes，and the glycosylation of exosomes may be as a circulating tumor marker. This review focuses on the physiological significance，clinical value and therapeutic prospect of exosomal glycosylation，and it introduces the exosome capture techniques，which provides methodological support for the further study of exosomal glycosylation. The study of exosomal glycosylation will provide a new strategy for the application of liquid biopsy in the accurate diagnosis，high risk warning and accurate management of malignant tumors.

Application of MALDI-TOF MS technology in clinical microbiological examination

DUAN Xuehan, WU Hua

2024, 39(4):  410-414.  DOI: 10.3969/j.issn.1673-8640.2024.04.017

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With the development of science and technology，the application of new technology in medical examination is more and more extensive. Compared with the complicated and time-consuming workflow of traditional microbial identification，matrix-assisted laser desorption ionization time-of-flight mass spectrometry（MALDI-TOF MS） has gradually become an indispensable microbial identification tool in clinical microbiological laboratories because of its rapid，sensitivity，simple，time-saving and high specificity. MALDI-TOF MS plays a role in isolate identification，drug resistance monitoring and mutation identification. Its identification relies on the database，and it is limited to a certain extent by the quality of clinical samples. MALDI-TOF MS will still have utilization space to be developed in clinical microbiological examination.