Table of Content

30 May 2022, Volume 37 Issue 5

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Original article

Effect of Qinhuang Formula on immune function of patients with diffuse large B-cell lymphoma after R-CHOP chemotherapy

RUAN Ming, LIN Han, WANG Jianbiao, LÜ Lingling, ZHU Wenhua, XU Jiayue, LI Weili, ZHU Weirong

2022, 37(5):  403-408.  DOI: 10.3969/j.issn.1673-8640.2022.05.001

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Objective To investigate the effect of Qinhuang Formula on immune function of patients with diffuse large B-cell lymphoma（DLBCL） after R-CHOP chemotherapy. Methods A total of 300 cases of DLBCL undergoing R-CHOP chemotherapy were analyzed retrospectively and were classified into 2 groups. The treatment group（150 cases） were treated with Qinhuang Formula after chemotherapy for 6 months at least,and the control group（150 cases） were not treated with traditional Chinese medicine after chemotherapy. T cell subsets（the absolute values of CD3⁺T,CD4⁺T and CD8⁺T cells,the ratio of CD4/CD8）,IgG,IgA and IgM were observed in the 2 groups before chemotherapy,3 months after chemotherapy,completing chemotherapy,3 months after completing chemotherapy,6 months after completing chemotherapy,1 year after completing chemotherapy and 2 years after completing chemotherapy. Results The absolute values of CD3⁺T,CD4⁺T cells,CD4/CD8 ratio,the levels of IgG,IgA and IgM were decreased after chemotherapy（P<0.05）. During follow-up period,the absolute value of CD4⁺T cells increased significantly half a year after completing chemotherapy（P<0.05）. IgG was higher at 2 years after completing chemotherapy（P<0.05）. The absolute values of CD3⁺T cells and the ratio of CD4/CD8 in treatment group at 1-2 years after completing chemotherapy returned to the levels before chemotherapy,while those of control group was still lower than those before chemotherapy（P<0.05）. Compared with 3 months after chemotherapy,the absolute values of CD3⁺T and CD4⁺ T cells and the ratio of CD4/CD8 at 2 years after completing chemotherapy in treatment group increased significantly than those in control group（P<0.05）. There was no statistical significance in the absolute value of CD8⁺T cells between the 2 groups at each time point after completing chemotherapy（P>0.05）. The levels of IgG,IgA and IgM in the 2 groups after completing chemotherapy were lower than those before chemotherapy（P<0.05）. IgA returned to pre-chemotherapy level at 1 year after completing chemotherapy,and IgG returned to pre-chemotherapy level at 2 years after completing chemotherapy. IgM level at 2 years after completing chemotherapy was still lower than that before chemotherapy（P<0.05）. There was no statistical significance in the increase of IgG,IgA and IgM between the 2 groups（P>0.05）. Conclusions Long-term immunosuppression is appeared after R-CHOP chemotherapy in patients with DLBCL. Qinhuang Formula can promote the immune function reconstruction of DLBCL patients after chemotherapy.

Expression level of lncRNA SNHG15 in gastric cancer and clinical significance

JI Ye, YUAN Xiaosun, ZHANG Lei, MA Huili, DONG Wei, LI Changsheng, ZHANG Jingwei, REN Zhonghai, ZHANG Yifei

2022, 37(5):  409-412.  DOI: 10.3969/j.issn.1673-8640.2022.05.002

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Objective To investigate the clinical significance of long non-coding RNA（lncRNA） small nuclear RNA host gene 15（SNHG15） in gastric cancer. Methods A total of 97 patients with gastric cancer who underwent elective radical surgery were enrolled. The clinical data of all the patients were collected,and real-time quantitative polymerase chain reaction（PCR） was used to determine the relative expression levels of SNHG15 in gastric cancer tissue and normal gastric mucosal tissue. All the patients were followed up for 5 years from the 1st day after the operation. Survival analysis was performed by Kaplan-Meier survival curve,and prognostic factors were analyzed by Cox proportional hazard model. Results The relative expression level of SNHG15 in gastric cancer tissue was higher than that in normal gastric mucosal tissue（P<0.001）. The differences of the relative expression levels of SNHG15 in gastric cancer tissues with different degree of differentiation,TNM stage,depth of invasion and lymph node metastasis had statistical significance（P<0.05）. Kaplan-Meier survival curve analysis showed that the average survival time and 5-year survival rate of patients in low expression（SNHG15≤2.18） group were higher than those in high expression（SNHG15>2.18） group（P<0.001）. Cox proportional hazard model analysis showed that poorly differentiation,TNM stage Ⅲ-Ⅳ,lymph node metastasis and the expression of SNHG15 were risk factors that affected the prognosis of gastric cancer patients [hazard ratios（HR） were 0.381,2.568,2.892 and 4.851]. Conclusions The expression of SNHG15 in gastric cancer tissues is increased significantly,and it is related to the prognosis of patients,or can be used as a potential marker for gastric cancer mechanism research and prognosis evaluation.

Relationship of mtDNA T16189C mutation and gestational diabetes mellitus

ZHANG Xiaoqin, ZHOU Yulan, FU Zhumei, HAN Xuexue, ZHAN Jie

2022, 37(5):  413-416.  DOI: 10.3969/j.issn.1673-8640.2022.05.003

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Objective To study the relationship between the mutation of mitochondrial DNA（mtDNA） T16189C and the risk of gestational diabetes mellitus（GDM）. Methods Totally,204 patients with GDM were enrolled as GDM group,and 220 healthy pregnant women were enrolled as control group. The general data of all the pregnant women,including age,gestational week,body mass index（BMI）,fasting blood glucose（FBG）,oral glucose tolerance test（OGTT） 1 h blood glucose,OGTT 2 h blood glucose,glycated hemoglobin A_1c（HbA_1c） and fasting insulin（FINS）,were collected. The homeostasis model assessment for insulin resistance（HOMA-IR） was calculated. The genotype of mtDNA 16189 mutation was determined by sequencing. Results The mutation rate of this site of GDM group was higher than that of control group（P<0.05）. The mtDNA T16189C mutation can increase the risk of GDM [odds ratio（OR）=1.36,95% confidence interval（CI） 1.020-1.823]. GDM patients were classified into carrying group and non-carrying group according to the mtDNA T16189C mutation. The levels of OGTT 1 h blood glucose,OGTT 2 h blood glucose,FINS and HOMA-IR in carrying group were higher than those in non-carrying group（P<0.05）. There was no statistical significance for FBG and HbA_1c between the 2 groups（P>0.05）. Conclusions The mtDNA T16189C mutation can increase the risk of GDM,and mtDNA T16189C mutation is risk to GDM.

Serum miR-21 and miR-135a expression levels in predicting the recurrence and metastasis of gastric cancer patients after chemotherapy

LIANG Chunfang, ZHU Kangning, ZHANG Qi

2022, 37(5):  417-422.  DOI: 10.3969/j.issn.1673-8640.2022.05.004

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Objective To investigate the predictive value of serum miR-21 and miR-135a expression in the recurrence and metastasis of gastric cancer patients after chemotherapy. Methods A total of 100 gastric cancer patients（gastric cancer group） who underwent chemotherapy were enrolled,and 50 healthy subjects were enrolled as control group. The clinical data of all the patients were collected. Follow-up started 1 month after chemotherapy,and the follow-up time was 24 months. The relative expression levels of serum miR-21 and miR-135a in control subjects and gastric cancer patients before chemotherapy and at 3,8,13,18,and 24 months after chemotherapy were determined. Receiver operating characteristic（ROC） curve was used to evaluate the efficiency of miR-21 and miR-135a in judging the prognosis of gastric cancer. Results Compared with control group,the relative expression of miR-21 in gastric cancer group before chemotherapy increased（P<0.05）,and the relative expression of miR-135a decreased（P<0.05）. The relative expression levels of serum miR-21 and miR-135a before chemotherapy were related to lymph node metastasis,differentiation degree and the depth of invasion in patients with gastric cancer（P<0.05）,but they were not related to age,sex,TNM stage and the presence or absence of distant metastasis（P>0.05）. The 3 cases of 100 gastric cancer patients were lost to follow-up,and 66 cases（68%） had recurrence or metastasis. The relative expression of serum miR-21 in patients with recurrence or metastasis after chemotherapy increased with the extension of follow-up time（P<0.05）,and the relative expression of serum miR-135a decreased with the extension of follow-up time（P<0.05）. ROC curve analysis results showed that the areas under curves（AUC） of miR-21 and miR-135a single determinations to judge the poor prognosis of gastric cancer were 0.789 and 0.826,respectively. Taking both positive results as combined determination positive,and taking both negative results as combined determination negative,the sensitivity and specificity of the combined determination of miR-21 and miR-135a in judging the prognosis of gastric cancer were 91.2% and 67.8%,respectively. Conclusions The expressions of miR-21 and miR-135a in serum of patients with gastric cancer are abnormal,and the expression level is closely related to lymph node metastasis,the degree of differentiation and the depth of invasion. The combined determination of serum miR-21 and miR-135a has a value in judging the recurrence or metastasis of gastric cancer after chemotherapy.

Correlation between admission blood calcium level and the prognosis of patients with hypertensive intracerebral hemorrhage

HUANG Liying, SAI Fangfang, ZHANG Kebei, MENG Chao

2022, 37(5):  423-428.  DOI: 10.3969/j.issn.1673-8640.2022.05.005

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Objective To investigate the correlation between admission blood calcium level and 30 d mortality rate and 6-month functional outcome in patients with hypertensive intracerebral hemorrhage. Methods The data of patient age,sex,complications,the National Institutes of Health Stroke Scale （NIHSS）,intracranial hematoma volume and location and clinical laboratory results from 107 patients with hypertensive intracerebral hemorrhage were analyzed retrospectively. The patients were classified into >2.75 mmol/L group（15 cases）,2.25-2.75 mmol/L group（63 cases）and <2.25 mmol/L group（29 cases）. A modified Rankin Scale（mRS）>2 denoted poor functional outcome at 6 months. The correlations between calcium level,30 d mortality rate and 6-month functional outcome in hypertensive intracerebral hemorrhage was evaluated by Logistic regression analysis. Receiver operating characteristic （ROC） curve was used to distinguish hypertensive intracerebral hemorrhage patients with favorable and unfavorable functional outcome. Results Compared with >2.75 mmol/L group and 2.25-2.75 mmol/L group,the patients in <2.25 mmol/L group were presented with significantly larger intracranial hematoma（P<0.05）,higher NIHSS score and mRS score（P<0.05）,and more early hematoma enlargement 30 d mortality cases（P<0.05）and there was no statistical significance among the 3 groups（P>0.05）. Calcium level at admission predicted 30 d mortality and 6-month functional outcome with areas under curves（AUC） of 0.74 and 0.71,respectively. Calcium levels at admission were correlated with hematoma volume and early hematoma enlargement [odds ratio（OR）=0.24,95% confidence interval（CI）0.11-0.69;OR=0.15,95%CI 0.08-0.87,respectively]. The OR of 30 d mortality and poor functional outcome at 6 months of <2.25 mmol/L group were 1.12 and 1.39 as compared with >2.75 mmol/L group. Conclusions A low calcium level at admission is correlated with 30 d mortality and poor 6-month functional outcome for patients with hypertensive intracerebral hemorrhage. Blood calcium level determination may conduce to patients' prognosis identification.

Role of fecal calprotectin in children's intestinal diseases

ZHAO Ying, XU Jin

2022, 37(5):  429-432.  DOI: 10.3969/j.issn.1673-8640.2022.05.006

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Objective To investigate the role of fecal calprotectin in children's inflammatory bowel disease（IBD）. Methods Totally,125 children with IBD（IBD group） were enrolled,there were 33 cases in the attack stage of Crohn's disease,31 cases in acute enteritis,41 cases in ulcerative colitis,20 cases in abdominal allergic purpura and 19 cases in the remission stage of Crohn's disease,and there were 33 healthy children who underwent physical examination as healthy control group. Fecal calprotectin levels of all the subjects were determined. Receiver operating characteristic（ROC） curve was used to evaluate the efficiency of fecal calprotectin in identifying the onset and remission stages of Crohn's disease. Results The fecal calprotectin levels in IBD group was higher than that in healthy control group（P<0.000 1）. The fecal calprotectin levels of children with different types of IBD were higher than those of healthy control group（P<0.000 1）,but there was no statistical significance in the fecal calprotectin levels of children with different types of IBD（P>0.05）. The fecal calprotectin level in Crohn's disease attack group was higher than that in remission group（P<0.000 1）. ROC curve analysis showed that the area under curve（AUC） of fecal calprotectin in identifying the onset and remission stages of Crohn's disease was 0.933 8,the optimal cut-off value was 59.91 μg/g,the sensitivity was 89.47%,and the specificity was 82.86%. Conclusions Fecal calprotectin can be used as an auxiliary diagnostic indicator of IBD and an identification indicator between attack and remission stages of Crohn's disease.

Characteristics of blastic plasmacytoid dendritic cell neoplasm

FANG Yinghui, WANG Weiguo, ZHANG Hua, GU Yan, ZHENG Fang, LI Liping

2022, 37(5):  433-437.  DOI: 10.3969/j.issn.1673-8640.2022.05.007

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Objective To report a case of blastic plasmacytoid dendritic cell neoplasm（BPDCN）,and to detail the characteristics of its clinical manifestation,cell morphology,immunophenotype and karyotype analysis,which may show new light on the diagnosis of the disease. Methods The case was diagnosed by its clinical manifestation,cell morphology and immunophenotype. Literature review was performed to reveal the clinical characteristics of BPDCN. Results The results of bone marrow smear demonstrated the presence of a dramatically active myeloid hyperplasia,and the presence of the blast cell was 37.5%,with the characteristic of median size,circular or elliptic or irregular nucleus,loose chromatin,0 to 3 nucleolus,dispersed gray-blue cytoplasm and vacuolation which was often observed at the edge of cytoplasm. Peroxidase（POX）staining showed the blast cells were negative,while periodic acid-schiff（PAS） staining displayed that they were positive. Flow cytometric analysis of the bone marrow revealed the presence of the abnormal cells was 13.15%,and they were strongly positive for HLA-DR,CD56,CD123,positive for CD33,CD7,CD4,partially positive for CD38,and negative for cCD3,cCD22,cMPO,CD3,CD10,CD34,CD117. The result of karyotype analysis was 46,XY（20）. The patient was diagnosed as BPDCN with the clinical manifestation,cell morphology and immunophenotype. Conclusions The characteristics of clinical manifestation,cell morphology and immunophenotype are of clinical significance to diagnose BPDCN,and the characteristics of immunophenotype is indispensable.

Clinical significance of von Willebrand factor determination in elderly patients with chronic heart failure complicated with lower extremity venous thrombosis

SONG Jinping, MA Jing, HUANG Guohong

2022, 37(5):  438-442.  DOI: 10.3969/j.issn.1673-8640.2022.05.008

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Objective To analyze the predictive value of von Willebrand factor（vWF） for venous thrombosis in elderly patients with chronic heart failure（CHF）. Methods A total of 96 elderly patients with CHF were enrolled. Among them,59 patients with CHF without lower extremity venous thrombosis were enrolled in disease control group,and 37 patients with chronic heart failure complicated with lower extremity venous thrombosis were enrolled in disease study group. Totally,50 healthy subjects over 60 years old during the same period were enrolled in healthy control group. The general clinical data were collected,and vWF activity,D-dimer and coagulation function related indexes were determined. Logistic regression analysis of influencing factors for venous thrombosis in patients with CHF was performed. The efficiency of plasma vWF activity on lower extremity venous thrombosis in elderly patients with CHF was evaluated by receiver operating characteristic（ROC） curve. Results The vWF activity was higher in disease research group than that in disease control group（P<0.05）. Logistic regression analysis showed that vWF activity was an independent risk factor for lower extremity venous thrombosis in patients with CHF. The diagnostic cut-off value of plasma vWF activity for CHF complicated with lower extremity venous thrombosis was 231.5%,the sensitivity was 78.6%,and the specificity was 82.6%. Conclusions Plasma vWF activity level has clinical value in assessing the risk of lower extremity venous thrombosis in elderly patients with CHF.

Level changes of GM and BG in patients with IPA before and after treatment in different immune states

HUANG Wanhong, MO Lida, LUO Xiaolu, LIN Yinghui, LIU Zeduan, CHEN Songlin

2022, 37(5):  443-446.  DOI: 10.3969/j.issn.1673-8640.2022.05.009

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Objective To analyze dynamically the changes of blood galactomannan（GM） and （1,3）-D-glucan（BG） levels in patients with invasive pulmonary aspergillosis（IPA） before and after treatment. Methods A total of 63 IPA patients with confirmed or clinical diagnosis were enrolled,and their peripheral blood GM and BG determinations and lymphocyte subset determinations were performed before treatment,1 month after treatment and 3 months after treatment. According to whether infected human immunodeficiency virus（HIV） （40 cases of HIV group and 23 cases of non-HIV group）or not and different CD4⁺ T cell levels（CD4⁺ T cell count<200 cells/μL in Group A,CD4⁺ T cell count 200-500 cells/μL in Group B,CD4⁺ T cell count >500 cells/μL in Group C）. The difference of BG,GM,CD3⁺,CD4⁺ and CD8⁺ T cells,CD4/CD8 was evaluated. Results GM and BG levels of 63 patients with IPA in 3 months after treatment were lower than those before treatment and in 1 month after trearment（P<0.05）. In HIV group,GM and BG levels were lower in 3 months after treatment than before treatment（P<0.05）,whereas in non-HIV group,GM levels were lower in 3 months after treatment than before treatment（P<0.05）. According to the classification of CD4⁺ T cells,the GM levels in Group A and Group B were lower than those before treatment and in 1 month after treatment and in 1 month after treatment（P<0.05）,and BG in Group A was lower in 3 months after treatment than before treatment（P<0.05）. The level of GM in Group C was lower than that in Group B before treatment and in 1 month after treatment（P<0.05）. Conclusions The combined determination of GM and BG plays a role for the diagnosis and antifungal treatment of IPA patients.

A multi-center evaluation of a domestic glutathione reductase determination kit

ZHU Yuqing, WANG Haibin, WANG Lianming, ZHAO Fang, FENG Pinning, ZHANG Peng, GUO Shuren, PENG Yibing, CHENG Liming, CHUAN Liangmin, WANG Hualiang

2022, 37(5):  449-455.  DOI: 10.3969/j.issn.1673-8640.2022.05.011

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Objective To investigate the performance of a domestic glutathione reductase（GR） determination kit through a multi-center evaluation. Methods Totally,9 clinical laboratories of tertiary hospitals located in different geographic regions of China formed a multi-center evaluation group. According to the same evaluation scheme,the repeatability,within-laboratory precision,between-run variations,linearities,the maximum dilution ratio and estimation of bias of a domestic GR determination kit were investigated,which were classified and compared. Results The repeatability and within-laboratory precision of the 9 laboratories met the requirements [less than 1/4 standard of external quality assessment（EQA）（5%） and 1/3 standard of EQA（6.67%）,respectively]. Except a low-level sample of one laboratory and a middle-level sample of another laboratory（>8%）,the between-run variations of all the other laboratories met the manufacturer's stated requirements. All the laboratories demonstrated the linear range claimed by the manufacturer. The recovery rates at maximum dilution ratio（5 times） of the 9 laboratories were 92.6%-106.3%. In the estimation of bias by using 40 patient samples compared with imported reagent,the correlation coefficients（r） were 0.997-1.000,the biases were less than 1/2 standard of EQA（10%）. There was a significant correlation among the results of the 9 laboratories in the imported group and the domestic group respectively（P<0.01）. The consistency of imported reagent and domestic reagent was good,and the difference of paired t-test was not significant statistically（P>0.05）,and the correlation coefficients（r） were 1.000（P<0.01）. The average coefficient of variation（CV） of imported reagent and domestic reagent was 6.48% and 6.68%,respectively. Pairwise paired t-test was performed on the results of instrument group A,B and C. There was statistical significance between group B and C for domestic reagent（P<0.01）. There was statistical significance between group A and C,B and C for imported reagent（P<0.01）. Conclusions The results of the multi-center evaluation show that the performance of the domestic GR reagent is good,and it can meet the clinical determination needs.

Establishment and evaluation for the determination of serum digitoxin as a candidate reference method

TANG Ningbo, LIU Guancai, LIU Xiaoyu, YANG Fengfan, GONG Zhimei, SUN Keqi

2022, 37(5):  456-462.  DOI: 10.3969/j.issn.1673-8640.2022.05.012

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Objective To establish a candidate reference method for the determination of serum digitoxin based on isotope dilution ultra high performance liquid chromatography tandem mass spectrometry（ID-UPLC-MS/MS）,and to evaluate the performance of this method. Methods The human serum was added with digitoxin-21,23,23-d₃,the protein was precipitated in 0.5 mol/L zinc sulfate solution. Then the samples were extracted with tert-butyl methyl ether,the upper layer was dried with nitrogen flow. The residual was reconstituted with mobile phase and analyzed by ID-UPLC-MS/MS. Serum digitoxin concentration was calculated by 5-point bracketing method. The ID-UPLC-MS/MS（matrix effect,specificity,carrying contamination,precision,accuracy）was validated according to the Clinical and Laboratory Standards Institute（CLSI）C62-A and CLSI EP15-A3. The uncertainty of the measurement of digitoxin by ID-UPLC-MS/MS. Results The ID-UPLC-MS/MS for the determination of digitoxin was validated over a concentration range of 2.8-94.9 nmol/L. The intra-batch coefficient of variation（CV） was 0.44%-1.98%,and the total CV was 0.41%-1.06%. The recoveries were 100.03%-100.48%. The limits of detection [signal-to-noise ratio（R_SN）=3] and quantitation（R_SN=10）were 0.032 and 0.095 nmol/L,respectively. According to RELA 2020,RELA-A and RELA-B 2020 relative biases of 0.62% and 0.29% were determined to be the uncertainty of 0.35 nmol/L. Conclusions The ID-UPLC-MS/MS established successfully for serum digitoxin has good precision,repeatability and accuracy,which can be used as a candidate reference method.

Role of miR-19b in human aortic valve interstitial cell calcification

YANG Meng, DING Hui

2022, 37(5):  466-471.  DOI: 10.3969/j.issn.1673-8640.2022.05.014

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Objective To investigate the role of miR-19b in human aortic valve interstitial cell（hVIC） calcification and its mechanism. Methods A calcification medium containing bone morphogenetic protein 2（BMP-2）,dexamethasone,anti-vitamin C and β-glycerophosphate was used to induce the calcification of hVIC,and Von Kossa staining was used to determine whether the model was constructed successfully. Untreated hVIC cells were used as control group. According to different transfection plasmids,hVIC cells were classified into miR-19b blank control（miR-NC） group,miR-19b overexpression group,SMAD4 overexpression group and miR-19b and SMAD4 overexpression（miR-19b+SMAD4 overexpression） group. The mRNA expression levels of miR-19b,Runt-related gene 2（RUNX 2）,osteocalcin（OC）,SMAD1,osteopontin（OPN） and SMAD4 in hVIC of each group were determined by fluorescent quantitative polymerase chain reaction（PCR）. The expression level of SMAD4 protein was determined by western blotting. The downstream target genes of miR-19b were predicted by StarBase database and verified by dual-luciferase reporter gene experiments. Results The hVIC after calcification induced overlap each other and showed an obvious aggregation-like growth trend,and the number of intracellular calcified nodules was increased（P<0.05）. The expression levels of RUNX2,OC,SMAD1 and OPN mRNA were increased（P<0.05）. The expression of miR-19b was down-regulated in the calcified hVIC model（P<0.05）,and the expression levels of osteogenic factors,hVIC intracell RUNX2 mRNA,OC mRNA,SMAD1 mRNA and OPN mRNA,in the model was decreased after the overexpression of miR-19b（P<0.05）. The results of dual luciferase reporter gene experiments confirmed that SMAD4 was the downstream target gene of miR-19b. The overexpression of SMAD4 up-regulated the expression of osteogenesis-related factors in calcified hVIC cells（P<0.05）,while the overexpression of miR-19b reversed the up-regulation effect of overexpression of SMAD4 on the expression of osteogenesis-related factors（P>0.05）. Conclusions The expression of miR-19b is down-regulated in the calcified hVIC model. By targeting the downstream target gene SMAD4 to regulate the calcification of hVIC,miR-19b and SMAD4 are potential therapeutic targets for treating aortic valve calcification diseases.

Influence of arginine metabolism of Mycolicibacterium smegmatis on the virulence of bacteria and mitochondrial function of alveolar epithelial cells

ZHOU Ziwei, LIN Chen, LI Yeyu, WANG Yuchen, ZHANG Lu

2022, 37(5):  472-476.  DOI: 10.3969/j.issn.1673-8640.2022.05.015

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Objective To investigate the influence of arginine metabolism on the virulence,pathogenicity and host physiology of Mycolicibacterium smegmatis（M. smeg）. Methods A M. smeg knockout isolate of argR was constructed. Wild-type M. smeg was used as control,and the growth ability of bacteria in vitro was determined. Real-time fluorescence quantitative polymerase chain reaction （RT-qPCR） was used to determine the expressions of fadD2,fadA5 and kstR. Epithelial cells A549 were infected with M. smeg. Cell activity was determined as well. UCP1 and DNM1L were determined by RT-qPCR. The physiological effect on host cells was observed. Results It was found that argR knockout increased the lipid storage capacity of bacteria and had a growth advantage in the medium with glucose as a single carbon source（P<0.001）. The synthesis of M. smeg mycoacid was up-regulated,and the virulence of epithelial cells A549 was increased. After the infection of epithelial cells A549 2 h,the cell entry rate was increased（P<0.001）,and the infection ability of epithelial cells A549 was improved. The deletion of argR affected cell viability by affecting its mitochondrial function. Conclusions The knockout of argR,a negative transcriptional regulator of arginine metabolism in M. smeg,enhances bacterial virulence and inhibits mitochondrial function in alveolar epithelial cells.

Research progress of electrochemical aptasensors for the determination of biomarkers

FENG Xueqing, JU Yi, LI Qing, JIN Zhonggan

2022, 37(5):  477-482.  DOI: 10.3969/j.issn.1673-8640.2022.05.016

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Aptamers have attracted more and more attention in clinical determination due to their specific recognition of disease biomarkers. At present,there have been many research reports,using aptamers to identify biomarkers to form determination method which has detectable signals. With the development of aptamer biosensors,its application prospects are also revealed. The combination of aptamers and functional nanomaterials can improve determination sensitivity and reduce determination limit. This review introduces the method of aptamer nanotechnology to construct electrochemical biosensors and how to improve the performance of biosensors. The application of biosensors in the determination of disease biomarkers is exemplified. The application prospects and challenges of aptamer biosensors are discussed in this review as well.

Research progress in the application of cerebrospinal fluid marker analysis in central nervous system leukemia

WANG Mian, YANG Jianmin, SONG Zhiqiang, TANG Gusheng

2022, 37(5):  483-488.  DOI: 10.3969/j.issn.1673-8640.2022.05.017

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In the recent years,with the improvement of treatment regimens and the application of new drugs,the rate of complete remission of leukemia has been significantly improved. However,many patients still experience relapse,the combination of central nervous system leukemia（CNSL） is an important cause,and the patients often have a poor prognosis. At present,the diagnosis of CNSL mainly relies on clinical laboratory tests and clinical symptoms and signs. This review focuses on not only the traditional tools used to determine clinical samples,including routine cerebrospinal fluid examination（cerebrospinal fluid pressure,cytological smear and biochemical tests）,cerebrospinal fluid flow cytometric immunophenotyping,but also an overview of other biomarkers,nucleic acid molecular diagnostics,metabolomics and other new technologies. These methodologies are expected to improve the sensitivity and specificity of the diagnosis of CNSL,to assist in the early diagnosis of central nervous system involvement,and to improve the clinical prognosis of patients by taking effective preventive and intervention measures.

Research progress of methicillin-resistant Staphylococcus aureus screening and decolonization to reduce surgical site infection

TANG Jin, PAN Yunqi, YANG Yumei, WU Qiong, WANG Jianqiang, LI Yungai, DI Jianzhong

2022, 37(5):  489-492.  DOI: 10.3969/j.issn.1673-8640.2022.05.018

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Surgical site infection is one of the most common healthcare-associated infections. Staphylococcus aureus remains the most common etiologic agent causing surgical site infection. The colonization of methicillin-resistant Staphylococcus aureus（MRSA） increases the risk of surgical site infection. Screening and decolonization of MRSA can reduce the rates of surgical site infection. The purpose of this review is to summarize the strategies to reduce surgical site infection due to MRSA focusing on decolonization.