1.Identification of metallothionein-associated macrophages and their predictive value for anti-TNF therapy response in inflammatory bowel disease
Gaoshi ZHOU ; Mudan ZHANG ; Minhu CHEN ; Shenghong ZHANG
Chinese Journal of Inflammatory Bowel Diseases 2025;09(3):199-210
Objective:To analyze the expression of zinc homeostasis-related genes and related cells in the intestinal mucosa of inflammatory bowel disease (IBD) patients at the single-cell level and to evaluate their value in predicting the response to anti-tumor necrosis factor (TNF) therapy in IBD patients.Methods:Single-cell RNA sequencing data from 75 ileal or colorectal biopsy samples, including those from patients with Crohn's disease (CD), ulcerative colitis (UC), and normal controls (NC), were collected from four gene expression omnibus (GEO) databases. Unsupervised clustering analysis in R language was employed to classify IBD cells, zinc homeostasis-related gene scores were used to assess the zinc homeostasis status of different cell clusters, and the cell clusters closely related to zinc homeostasis-related genes, namely metallothionein-associated macrophages (MT Mph), were identified. Then the colon tissues from IBD patients and healthy individuals treated at the First Affiliated Hospital of Sun Yat-sen University were collected for immunofluorescence (IF) staining to compare the differences in MT Mph numbers between IBD and NC tissues. To further explore the function and origins of MT Mph, the characteristic genes of MT Mph and non- metallothionein-associated macrophages (non-MT Mph) from database were compared, the Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis was further used to enrich the characteristic genes, cell communication analysis was used to investigate the communication mechanisms between MT Mph and different cells, Quasi-time sequence was used to explore the origin of MT Mph and related signaling pathways, and the differences in transcription factors among monocytes, MT Mph and non-MT Mph were analyzed in R language. Single sample gene set enrichment analysis (ssGSEA) was used to evaluate the expression of MT Mph characteristic genes, and ssGSEA combined with the response to anti-TNF were used to construct the model in order to explore the predictive value of MT Mph characteristic genes for the response to anti-TNF therapy in IBD patients.Results:IBD cells were clustered and annotated into seven major cell clusters, namely T cells, B cells, plasma cells, myeloid cells, fibroblasts, endothelial cells, and epithelial cells. The results of zinc homeostasis-related gene scores showed that the scores of IBD myeloid cells were higher than those of the NC group. Myeloid cells could be divided into monocytes, macrophages, neutrophils, and dendritic cells. Based on the expression of zinc homeostasis genes, especially the high expression of metallothionein genes, the macrophages were divided into MT Mph and non-MT Mph, and the number of MT Mph in the IBD intestine was significantly increased compared to the NC group. IF validation showed that the number of CD68 +MT1G + cells (MT Mph) in both UC and CD were significantly higher than that in the NC group under per high-power field of view (UC vs. NC: 30.80 ± 7.29 vs. 9.80 ± 1.80, P < 0.001; CD vs. NC: 36.00 ± 9.30 vs. 9.80 ± 1.80, P < 0.001). KEGG pathway enrichment analysis revealed that the differential genes of MT Mph were enriched in key genes of inflammation-related pathways, especially the TNF signaling pathway. Cell communication analysis showed that the TNF signaling pathway between MT Mph and other cells in IBD was significantly enhanced compared to NC. Quasi-time sequence analysis results showed that monocytes could differentiate into MT Mph, and the expression of metallothionein genes ( MT2A, MT1X, MT1H and MT1G) was significantly upregulated during the differentiation process. Transcription factor analysis showed that the transcription factors SMARCB1 and ZMYND8 of MT Mph were significantly higher than those of monocytes, and the classical inflammatory transcription factors HIF1A, STAT3, and NFKB1 were significantly higher than those of non-MT Mph. Prediction models for CD and UC were constructed respectively based on ssGSEA and TNF treatment response [CD: area under the curve (AUC) = 0.966, P < 0.01; AUC = 0.793, P < 0.01]. Validation results showed that the model could not predict the response of CD and UC patients to vedolizumab therapy (both P > 0.05). Conclusions:There is a zinc homeostasis imbalance in IBD intestine, and MT Mph are a group of cells with high expression of zinc homeostasis-related genes, which are closely related to the TNF inflammatory pathway. The prediction model constructed based on the characteristic genes of MT Mph may be able to predict the response to anti-TNF therapy in IBD.
2.Identification of metallothionein-associated macrophages and their predictive value for anti-TNF therapy response in inflammatory bowel disease
Gaoshi ZHOU ; Mudan ZHANG ; Minhu CHEN ; Shenghong ZHANG
Chinese Journal of Inflammatory Bowel Diseases 2025;09(3):199-210
Objective:To analyze the expression of zinc homeostasis-related genes and related cells in the intestinal mucosa of inflammatory bowel disease (IBD) patients at the single-cell level and to evaluate their value in predicting the response to anti-tumor necrosis factor (TNF) therapy in IBD patients.Methods:Single-cell RNA sequencing data from 75 ileal or colorectal biopsy samples, including those from patients with Crohn's disease (CD), ulcerative colitis (UC), and normal controls (NC), were collected from four gene expression omnibus (GEO) databases. Unsupervised clustering analysis in R language was employed to classify IBD cells, zinc homeostasis-related gene scores were used to assess the zinc homeostasis status of different cell clusters, and the cell clusters closely related to zinc homeostasis-related genes, namely metallothionein-associated macrophages (MT Mph), were identified. Then the colon tissues from IBD patients and healthy individuals treated at the First Affiliated Hospital of Sun Yat-sen University were collected for immunofluorescence (IF) staining to compare the differences in MT Mph numbers between IBD and NC tissues. To further explore the function and origins of MT Mph, the characteristic genes of MT Mph and non- metallothionein-associated macrophages (non-MT Mph) from database were compared, the Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis was further used to enrich the characteristic genes, cell communication analysis was used to investigate the communication mechanisms between MT Mph and different cells, Quasi-time sequence was used to explore the origin of MT Mph and related signaling pathways, and the differences in transcription factors among monocytes, MT Mph and non-MT Mph were analyzed in R language. Single sample gene set enrichment analysis (ssGSEA) was used to evaluate the expression of MT Mph characteristic genes, and ssGSEA combined with the response to anti-TNF were used to construct the model in order to explore the predictive value of MT Mph characteristic genes for the response to anti-TNF therapy in IBD patients.Results:IBD cells were clustered and annotated into seven major cell clusters, namely T cells, B cells, plasma cells, myeloid cells, fibroblasts, endothelial cells, and epithelial cells. The results of zinc homeostasis-related gene scores showed that the scores of IBD myeloid cells were higher than those of the NC group. Myeloid cells could be divided into monocytes, macrophages, neutrophils, and dendritic cells. Based on the expression of zinc homeostasis genes, especially the high expression of metallothionein genes, the macrophages were divided into MT Mph and non-MT Mph, and the number of MT Mph in the IBD intestine was significantly increased compared to the NC group. IF validation showed that the number of CD68 +MT1G + cells (MT Mph) in both UC and CD were significantly higher than that in the NC group under per high-power field of view (UC vs. NC: 30.80 ± 7.29 vs. 9.80 ± 1.80, P < 0.001; CD vs. NC: 36.00 ± 9.30 vs. 9.80 ± 1.80, P < 0.001). KEGG pathway enrichment analysis revealed that the differential genes of MT Mph were enriched in key genes of inflammation-related pathways, especially the TNF signaling pathway. Cell communication analysis showed that the TNF signaling pathway between MT Mph and other cells in IBD was significantly enhanced compared to NC. Quasi-time sequence analysis results showed that monocytes could differentiate into MT Mph, and the expression of metallothionein genes ( MT2A, MT1X, MT1H and MT1G) was significantly upregulated during the differentiation process. Transcription factor analysis showed that the transcription factors SMARCB1 and ZMYND8 of MT Mph were significantly higher than those of monocytes, and the classical inflammatory transcription factors HIF1A, STAT3, and NFKB1 were significantly higher than those of non-MT Mph. Prediction models for CD and UC were constructed respectively based on ssGSEA and TNF treatment response [CD: area under the curve (AUC) = 0.966, P < 0.01; AUC = 0.793, P < 0.01]. Validation results showed that the model could not predict the response of CD and UC patients to vedolizumab therapy (both P > 0.05). Conclusions:There is a zinc homeostasis imbalance in IBD intestine, and MT Mph are a group of cells with high expression of zinc homeostasis-related genes, which are closely related to the TNF inflammatory pathway. The prediction model constructed based on the characteristic genes of MT Mph may be able to predict the response to anti-TNF therapy in IBD.
3.Effect of induction and maintenance of intravenous or inhalation anesthesia on respiratory system adverse events in children after extubation
Chongqing Medicine 2024;53(20):3108-3112,3119
Objective To observe the effects of intravenous or inhalation anesthesia on perioperative re-spiratory system adverse events(PRAE)in children after extubation.Methods A total of 100 children pa-tients with elective tonsillectomy or adenoidectomy under general anesthesia endotracheal intubation in this hospital from July to December 2023 were selected as the study subjects and divided into the intravenous anes-thesia group and inhalation anesthesia group according to the random number table method,50 cases in each group.The intravenous anesthesia group used propofol for venous induction and maintenance,while the inha-lation anesthesia group used sevoflurane inhalation for induction and maintenance.The incidence rate of PRAE after extubation,anesthetic maintenance stage quality,incidence rates of postoperative nausea,vomiting and agitation were recorded in both groups.Results Compared with the inspiration anesthesia group,the PRAE incidence rate(16.0%vs.42.0%),hypoxia incidence rate(14.0%vs.38.0%)and pediatric anaesthesia e-mergence delirium(PAED)score[5(2,8)points vs.6(4,11)points]in the venous anesthesia group were low-er,the extubation time was longer[(19.6±10.6)min vs.(14.9±8.9)min],postanesthesia care unit(PACU)stay time was shorter[(25.4±5.2)min vs.(27.9±6.4)min],the differences were statistically significant(P<0.05).The proportions of intubation time,intubation times>once and the swallow and limb activity dur-ing anesthetic maintenance had no statistical difference between the two groups(P>0.05).The heart rate had no statistical difference among 4 time points of before and after induction,after intubation and after extu-bation(P>0.05).Compared with before induction,the heart rate after extubation was significantly increased,and the difference was statistically significant(P<0.05).Conclusion The incidence rate of PARE in pediatric ve-nous anesthesia is lower.
4.Labeling of carcinoembryonic antigen-specific CAR-T cells with superparamagnetic iron oxide nanoparticles and in vitro magnetic resonance imaging
Kungao HE ; Bo JIANG ; Mudan GUO ; Guiling WANG ; En ZHANG ; Doudou XU
Journal of Army Medical University 2024;46(17):1951-1958
Objective To use superparamagnetic iron oxide nanoparticles(SPIONs)to label chimeric antigen receptor(CAR)T cells targeting carcinoembryonic antigen(CEA),and perform magnetic resonance imaging(MRI)to real time trace CEA CAR-T cells in vivo.Methods Appropriate amount of ferumoxytol,heparin sodium and protamine sulfate were mixed at high(ferumoxytol 100 μg/mL,heparin sodium 4 IU/mL,protamine sulfate 120 μg/mL),medium(ferumoxytol 50 μg/mL,heparin sodium 2 IU/mL,protamine sulfate 60 μg/mL),and low(ferumoxytol 25 μg/mL,heparin sodium 1 IU/mL,protamine sulfate 30 μg/mL)concentrations to form a SPIONs complex ferumoxytol/heparin/protamine(FHP),and then co-incubated with CEA CAR-T cells for cell labeling.The biocompatibility of FHP was detected by CCK-8 assay,EdU assay and flow cytometry.The uptake of FHP was detected by Prussian blue staining,and SPIONs content in the cells was quantitatively detected by inductively coupled plasma-mass spectrometry(ICP-MS).Flow cytometry was used to detect the lytic effect of FHP-labeled CEA CAR-T cells on tumor cells,and MRI was employed to scan FHP-labeled CEA CAR-T cells.Results FHP at high,medium,and low concentrations had no significant effect on the activity of CEA CAR-T cells,with cell activity above 100%determined by CCK-8 assay.DNA proliferation was above 94.3%in EdU assays.Prussian blue staining showed that CEA CAR-T cells could take FHP up,with the uptake increased with the increment of FHP concentration.ICP-MS showed that the intracellular Fe content was 440.23±189.36 ng/mL.Tumor cell killing experiment showed that FHP-labeled CEA CAR-T cells had excellent killing capability against tumor cells.MRI scans indicated that T2WI signals of FHP-labeled CEA CAR-T cells were significantly reduced with increasing FHP concentration(P<0.01).Conclusion SPIONs complex FHP shows good biocompatibility and can effectively label CEA CAR-T cells.SPIONs complex FHP can be used as a magnetic marker for CEA CAR-T cells and a feasible MRI tracer for clinical application.
5.Application of magnetic compression anastomosis to colorectal anastomosis and reconstruction
Jing LI ; Guifang LU ; Miaomiao ZHANG ; Shiqi LIU ; Xiaopeng YAN ; Feng MA ; Xiaoyang REN ; Xuejun SUN ; Yi LYU ; Shuixiang HE ; Mudan REN
Chinese Journal of Digestive Endoscopy 2023;40(5):397-400
Clinical cases treated by magnetic compression anastomosis (MCA) for different causes and types of intestinal stenosis/ atresia to successfully achieve intestinal recanalization were reviewed, so as to explore the clinical application of MCA. From May 2019 to August 2022, 4 patients underwent colorectal MCA for intestinal recanalization in the First Affiliated Hospital of Xi'an Jiaotong University and Northwest Women and Children's Hospital. All operations went well, and the intestinal anastomosis was recanalized. The magnetic ring was discharged in 7-15 days, and the postoperative colonoscopy or radiography showed that the anastomosis was intact. MCA can be used to treat different types of colorectal stenosis and atresia due to different reasons, and can also be used to assist intestinal anastomosis in colorectal surgery.
6.Risk factors of cardiovascular and cerebrovascular diseases in young patients with hyperhomocysteinemia
Mudan FENG ; Yu XIA ; Ming SUN ; Yuzhen ZHANG ; Xia CHENG ; Yuanbin WU ; Yuhong WANG
Chinese Journal of Primary Medicine and Pharmacy 2023;30(6):881-884
Objective:To investigate the risk factors of cardiovascular and cerebrovascular diseases in young patients with hyperhomocysteinemia.Methods:A total of 260 patients younger than 45 years old who received treatment at the Department of Emergency, Seventh Medical Center, Chinese PLA General Hospital from January 2018 to January 2019 were included in this study. Among these patients, 126 patients with serum homocysteine levels ≥ 15.0 μmol/L were included in the hyperhomocysteinemia group, and 134 patients with serum homocysteine levels < 15.0 μmol/L were included in the control group. Height, body weight, body mass index, blood pressure, homocysteine, fasting blood glucose, blood uric acid, total cholesterol, triacylglycerol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and N-terminal B-type natriuretic peptide were determined in each group. Changes in risk factors of cardiovascular and cerebrovascular diseases were compared between the hyperhomocysteinemia and control groups.Results:There were significant differences in body mass index [(26.42 ± 3.54) kg/m 2vs. (22.14 ± 3.22) kg/m 2, t = 10.21, P = 0.016], blood uric acid [(308.71 ± 78.44) μmol/L vs. (285.05 ± 92.09) μmol/L, t = 2.22, P = 0.027], the incidence of coronary heart disease (73/126 vs. 61/134, χ2 = 4.00, P = 0.045) and the incidence of stroke (19/126 vs. 6/134, χ2 = 8.39, P = 0.004) between the hyperhomocysteinemia and control groups. There were no significant differences in fasting blood glucose, blood lipid level, N-terminal B-type natriuretic peptide, and the incidences of diabetes mellitus and hypertension between the two groups (all P > 0.05). Conclusion:The related risk factors of cardiovascular and cerebrovascular diseases increase in young patients with hyperhomocysteinemia. The incidences of coronary heart disease and stroke are very high, and therefore timely intervention should be carried out.
7.Effects of hsa_circ_0045943 targeting miR-106a on the biological characteristics of gastric cancer cells
Jingwei MA ; Ning ZHANG ; Meng ZHU ; Xinlan LU ; Zhiyong ZHANG ; Mudan REN
Journal of Xi'an Jiaotong University(Medical Sciences) 2022;43(4):509-515
【Objective】 To investigate the effects of hsa_circ_0045943 targeting miR-106a on the biological characteristics of gastric cancer cells and its mechanism. 【Methods】 Human gastric cancer cells MKN-45, AGS and gastric mucosal epithelial cells GES-1 were cultured; circ_0045943 was detected by real-time polymerase chain reaction. The overexpression and silencing of circ_0045943 adenovirus vectors OE-circRNA and sh-circRNA together with their negative controls OE-NC and sh-NC were constructed and transfected; CCK-8 method was used to detect the proliferation activity of AGS cells after overexpression and silencing of circ_0045943; TUNEL method was used to detect the cell apoptosis; transwell assay was used to detect the cell migration and invasion; and would healing assay was used to detect the cell migration. Starbase database screened the binding site of miR-106a and circ_0045943. Real-time PCR was used to detect the expression of miR-106a, and the expression of circ_0045943 and the changes of miR-106a after the treatment of OE-circRNA and sh-circRNA. 【Results】 Real-time PCR showed that the expression of circ_0045943 decreased in gastric cancer cells MKN-45 and AGS compared to GES-1 (P
8.Mechanism of sinomenine in regulating M2 macrophage polarization induced by gastric cancer cells
Yifei CHEN ; Mudan REN ; Xinlan LU ; Guifang LU ; Dan ZHANG ; Yan ZHAO ; Yarui LI ; Dan GUO ; Shuixiang HE
Journal of Xi'an Jiaotong University(Medical Sciences) 2022;43(3):436-443
【Objective】 To study the role and mechanism of sinomenine in the macrophage polarization induced by gastric cancer cells. 【Methods】 Sinomenine was added to gastric cancer cells BGC-823 and MKN-45, cell viability was measured by CCK-8, cell proliferation was measured by colony formation experiment, Co-culture and Transwell cell migration experiments were used to evaluate the recruitment and polarization of macrophages by sinomenine, flow cytometry was used to evaluate the polarization of macrophages, and qRT-PCR and Western blot were used to detect the expression of gene RNA and protein levels. 【Results】 Sinomenine could inhibit the proliferation of gastric cancer cells and the recruitment of gastric cancer cells to macrophages, thus promoting macrophage M2 polarization. It simultaneously inhibited the expression of STAT6 as well as the expression and phosphorylation of C/EBPβ. When STAT6 is overexpressed, it could reduce these inhibitory effects of sinomenine on gastric cancer cells. Further research found that STAT6 mediated the secretion of IL-6 by gastric cancer cells, which was the cause of sinomenine-mediated macrophage recruitment and M2 polarization. 【Conclusion】 The natural drug sinomenine has a good tumor-suppressing ability against gastric cancer, directly inhibits the survival and migration of gastric cancer cells, and inhibits the expression of IL-6 and the M2 phenotype in the tumor microenvironment, reshapes the tumor environment, and reduces the risk of M2 type macrophages for gastric cancer tumors.
9.Infection control measures and practice in gastrointestinal endoscopy departments during the novel coronavirus infection period
Xu ZHANG ; Qian ZHAO ; Xiaona KANG ; Xinlan LU ; Mudan REN ; Juan ZHANG ; Li REN ; Suhong ZHANG ; Zhiyong ZHANG ; Yan YIN ; Junwang XU ; Shuixiang HE ; Guifang LU
Journal of Xi'an Jiaotong University(Medical Sciences) 2021;42(1):128-131,151
【Objective】 To explore and evaluate infection control measures of preventing cross-contamination of novel coronavirus during gastrointestinal endoscopy treatment. 【Methods】 According to the hospital’s infection control requirements and related documents, infection control measures were formulated and implemented by combining with our actual clinical situation, including the management of the endoscope room, management and protection of patients and endoscopists. Then, we evaluated the effect of these measures. 【Results】 From January 25 to March 10, 2020, a total of 71 patients (53 males and 18 females) completed gastrointestinal endoscopy treatment, with an average age of 54 years (28-81 years). There were 36 (50.7%) cases of emergency treatment. All patients had been kept in quarantine for about 14 days (24±13), and no cross-contamination of novel coronavirus occurred. 【Conclusion】 During the novel coronavirus infection epidemic period, reasonable and effective measures should be taken to minimize the risk of infection in doctors and patients. The endoscope center should strengthen preoperative screening and management of patients, master indications of endoscopic procedures, complete endoscopists’ management and protection work, strictly follow the specifications of sterilizing gastrointestinal endoscopes, and construct the layout of "three zones and two passages".
10.Correlation study between adiponectin and inflammatory markers, nutritional status and severe acute exacerbation in patients with acute exacerbation of chronic obstructive pulmonary disease
Mudan HE ; Min ZHANG ; Yulan ZHANG ; Peng WANG ; Xingjing LI
Chinese Journal of Postgraduates of Medicine 2020;43(11):1000-1004
Objective:To investigate the correlation between serum adiponectin (APN) and inflammatory markers, nutritional status and severe acute exacerbation in late stage in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD).Methods:One hundred and sixty-three male patients with AECOPD from July 2017 to April 2019 in Wusong Hospital of Baoshan District, Shanghai City were selected. The patients were divided into low APN group (serum APN ≤ 102.38 μg/L, 97 cases) and high APN group (serum APN>102.38 μg/L, 66 cases). The interleukin (IL)-6, IL-8, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), hemoglobin, albumin, prealbumin, body mass index (BMI) and number of severe acute exacerbation in 1 year′ follow-up were compared between 2 groups, and the correlation between APN and inflammatory markers, nutritional status and number of severe acute exacerbation were analyzed.Results:The BMI, hemoglobin, prealbumin and albumin in low APN group were significantly higher than those in high APN group: (23.20 ± 3.26) kg/m 2 vs. (20.77 ± 3.78) kg/m 2, (133.23 ± 17.13) g/L vs. (125.68 ± 20.83) g/L, (185.36 ± 57.60) g/L vs. (148.83 ± 64.79) g/L, (37.09 ± 4.06) g/L vs. (33.77 ± 5.86) g/L, and there were statistical differences ( P<0.01 or<0.05); there were no statistical differences in total protein, IL-6, IL-8, CRP and ESR between 2 groups ( P>0.05). APN showed negative correlation with BMI, hemoglobin, albumin and prealbumin ( r = -0.42, -0.28, -0.33 and -0.31; P<0.01); there was no correlation with IL-6, IL-8, ESR and CRP ( P>0.05). The rate of 4 times of severe acute exacerbation in 1 year′ follow-up in low APN group was significantly lower than that in high APN group: 8.25% (8/97) vs. 22.73% (15/66), and there was statistical difference ( χ2 = 6.79, P<0.05). Conclusions:The AECOPD patients with low APN have better nutritional status and less exacerbation. Serum APN can be used as a predictive index of severe acute exacerbation in AECOPD patients

Result Analysis
Print
Save
E-mail