1.Hepatic T cell subtypes and functional analysis among alveolar echinococcosis patients using single-cell RNA sequencing
Si CHEN ; Xiangqian WANG ; Wanzhong JIA ; Qigang CAI ; Xueyong ZHANG ; Qiang ZHANG ; Haibo ZHENG ; Linghong ZHU ; Bing LI ; Wei WANG ; Xiumin HAN
Chinese Journal of Schistosomiasis Control 2024;36(5):481-493
Objective To investigate T cell subtypes and their functions in liver immune microenvironments among patients with alveolar echinococcosis (AE) using single-cell RNA sequencing (scRNA-seq). Methods Four AE patients that were admitted to Qinghai Provincial People’s Hospital in 2023 for hepatic surgery for the first time were enrolled, and liver specimens were sampled 1 cm (peri-lesion, PL group) and > 5 cm from AE lesions (distal lesion, DL group) among each patient. Finally, a total of eight liver specimens were sampled from four AE patients for scRNA-seq analysis. Genome and transcriptome data of liver specimens were processed using the software Cell Ranger and R package. Differentially expressed genes (DEGs) and their biological functions were analyzed using gene ontology (GO) enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis, and the primary intercellular communication patterns and interaction mechanisms were identified among T cell subtypes in liver specimens using the CellChat package. In addition, the developmental stages of T cells were subjected to trajectory analysis with the monocle package to investigate the expression of genes associated with cell growth and tumor transformation, and to predict the developmental trajectories of T cells. Results All four AE patients were female, with a mean age of (25.00 ± 9.06) years, and there were three cases from Jiuzhi County, Golog Tibetan Autonomous Prefecture and one case from Chengduo County, Yushu Tibetan Autonomous Prefecture, Qinghai Province. The viability of single-cell samples from eight liver specimens was 90.41% to 96.33%, and a total of 81 763 cells were analyzed, with 19 cell types annotated. Of these cell types, 13 were immune cells (87.60%), and T cells (33.13%), neutrophils (15.40%), and natural killer cells (11.92%) were the three most common cell types. Re-clustering of 27 752 T cells and proliferative T cells identified 10 distinct T cell subtypes, with CD8+ cytotoxic T cells (23.43%), CD8+ naive T cells (12.80%), and CD4+ effector memory T cells (17.73%) as dominant cell types. The proportions of T helper 2 (Th2) cells (5.19% vs. 3.63%; χ2 = 38.35, P < 0.01) and CD4+ effector memory T cells (21.59% vs. 13.67%; χ2 = 244.70, P < 0.01) were significantly higher in liver specimens in the PL group than in the DL group, and the proportion of CD4+ helper T cells was significantly lower in the PL group than in the DL group (7.50% vs. 14.75%; χ2 = 330.52, P < 0.01). KEGG pathway analysis revealed that Th2 cells were significantly enriched in cell apoptosis and multiple cancer-associated pathways, and CD4+ effector memory T cells were significantly enriched in the regulation of cytokines and chronic inflammation, while CD4+ helper T cells were significantly enriched in immune responses regulation. Trajectory analysis of T cells showed that CD4+ helper T cells were at an earlier developmental stage relative to Th2 cells and CD4+ effector memory T cells, and the expression of inhibitor of DNA binding 3 (ID3), thioredoxin interacting protein (TXNIP), Bcl2-associated athanogene 3 (BAG3) and heat shock protein family B (small) member 1 (HSPB1) genes appeared a tendency towards a decline over time. Conclusions CD4+ effector memory T cells and CD8+ cytotoxic T cells are primary interacting cells in the liver specimens of AE patients. Reduced expression of Th2 cells and CD4+ helper T cells contributes to an inhibitory immune microenvironment, which promotes immune evasion by Echinococcus multilocularis, and Th2 cells are significantly enriched in multiple cancer-associated pathways, which may be linked to the invasive growth of E. multilocularis.
2.Risk factors and risk prediction model for coronary atherosclerotic heart disease in Xining Area
Xiaomin DAI ; Bo CHEN ; Na HAN ; Huicong XI
Journal of Public Health and Preventive Medicine 2024;35(6):109-112
Objective To screen the risk factors and risk prediction model for coronary atherosclerotic heart disease in Xining area. Methods Five hundred and eighteen patients with coronary atherosclerotic heart disease who attended the Qinghai Specialized Hospital for Cardiovascular and Cerebrovascular Diseases and Cerebrovascular Diseases from May 2018 to September 2022 were selected as the observation group, and another 421 patients with non-coronary heart disease were set as control group. The general information of patients were collected. The risk factors affecting the development of coronary heart disease were screened using Logistic regression analysis, and a risk prediction model was constructed, then receiver operating characteristic (ROC) curve was plotted to validate the predictive value of risk model. Results The gender ratio and smoking history yielded no statistical difference between two groups (P>0.05), but statistical difference was found in age, body mass index (BMI), diabetes history, hypertension history, smoking history and family history between two groups (P<0.05). Serum levels of total cholesterol (TC), low-density lipoprotein (LDL) and uric acid (UA) were all higher than the control group, and serum high-density lipoprotein (HDL) level was lower than the control group, with statistical difference (all P<0.05). Multivariate Logistic regression analysis denoted that age, BMI, diabetes history, hypertension history, smoking history, family history, TC, LDL, and UA were risk factors for the development of coronary heart disease (P<0.05). ROC curve analysis yielded an AUC of 0.890 for the risk model, with a sensitivity and specificity of 72.03% and 91.14%, respectively. Conclusion Patient's age, BMI, and the presence of diabetes mellitus, hypertension and smoking, family history, abnormal blood lipid profiles, and abnormal blood uric acid are all risk factors for the development of coronary heart disease, and the risk model constructed on the basis of the above risk factors has a high degree of sensitivity and specificity, which is of great value in accurately evaluating the risk of coronary heart disease.
3.Adjustable external fixation traction combined with arthroscopic microfracture for treatment of osteochondral lesions of the talus
Huaqing YANG ; Yaohua ZHANG ; Qiang LI ; Liang LIU ; Hailin XU ; Qinghai HAN ; Dianzhao ZHAO ; Yun YANG ; Qichang YANG
Chinese Journal of Orthopaedic Trauma 2024;26(3):241-246
Objective:To investigate the clinical effects of adjustable external fixation traction combined with arthroscopic microfracture in the treatment of osteochondral lesions of the talus (OLT).Methods:A retrospective study was conducted to analyze the data of 27 OLT patients who had been treated at Department of Orthopedics, Beijing Rehabilitation Hospital from May 2017 to March 2022. There were 16 males and 11 females, aged (32.4±7.2) years. Lesion site: 23 medial and 4 lateral cases; Hepple staging: 7 cases at stage Ⅰ, 15 cases at stage Ⅱ, and 5 cases at stage Ⅲ; disease duration: (10.6±3.3) months. All the patients were treated by adjustable external fixation traction combined with arthroscopic microfracture. Recorded were the patients' visual analogue scale (VAS) pain scores and American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot scores at 6 months and 12 months after surgery, levels of interleukin-1 (IL-1), interleukin-6 (IL-6) and tumour necrosis factor- α (TNF- α) at 1 month after surgery, lesion area at 12 months after surgery, and incidence of complications. Results:The follow-up time for this cohort was (16.2±6.7) months. The AOFAS score was (61.52±6.75) points before surgery, (84.15±5.56) points at 6 months after surgery and (95.67±4.30) points at 12 months after surgery. The VAS score was (5.88±1.02) points before surgery, (2.12±0.48) points at 6 months after surgery and (0.66±0.36) points at 12 months after surgery. The two-by-two comparisons between the 3 time points for the above items were statistically significant ( P<0.05). IL-1 was (32.37±6.64) pg/mL, IL-6 (34.04±7.12) pg/mL, and TNF- α (17.89±4.96) ng/L at 1 month after surgery in the 27 patients, all of which were significantly lower than their preoperative levels [(96.63±14.80) pg/mL, (102.33±20.42) pg/mL, and (54.48±9.33) ng/L] ( P<0.05). The lesion area was (28.66±6.52) mm 2 at 12 months after surgery, significantly smaller than the value before surgery [(128.52±11.32) mm 2] ( P<0.05). Infection at the adjustable external fixation needle track occurred in 1 patient and lower limb thrombosis in 2 patients. Conclusion:In the treatment of OLT, adjustable external fixation and traction combined with arthroscopic microfracture can achieve satisfactory results and improve symptoms for the patients.
4.Research progress on HEG1 in cardiovascular generation and tumor development
Yaoyao WANG ; Xinjuan YU ; Jiahui WANG ; Qinghai LI ; Wei HAN
Chinese Journal of Preventive Medicine 2024;58(1):136-140
Heart development protein with EGF-like domains 1 (HEG1) is a novel mucin-like membrane protein with a long O-glycosylation region and EGF domain. HEG1 plays critical roles in embryo development and cardiogenesis, and is closely related to the occurrence and progression of malignant tumors. Here this article demonstrates the research progress on HEG1 in cardiovascular formation and tumor development in recent years, to inspire new ideas for the pathogenesis, diagnosis and treatment of related diseases.
5.Therapeutic effect and mechanism of Shiwei-Ruxiang-capsule on rheumatoid arthritis based on intestinal flora and metabolomics
Yu-han GAO ; Hai-juan CHEN ; Yong-gui MA ; Jun SHANG ; Guo-yan ZHANG ; Wen-jian ZHANG
Acta Pharmaceutica Sinica 2024;59(12):3304-3314
Shiwei-Ruxiang-capsule (SWRXC) is a classic formulation widely used in the treatment of rheumatoid arthritis (RA). The study used liquid chromatography-tandem mass spectrometry (LC-MS/MS) serum untargeted metabolomics and high-throughput 16S rRNA gene sequencing association analysis to elucidate the mechanism of action of SWRXC for the treatment of Freund's complete adjuvant-induced RA. The results showed that SWRXC significantly improved symptoms and reduced serum cytokine levels in RA rats. Based on LC-MS/MS technology, metabolomics identified tryptophan metabolism, nucleotide metabolism and purine metabolism as the most relevant pathways for treatment. In addition, 16S rRNA sequencing results showed that SWRXC could ameliorate RA-induced intestinal microbial oncogenesis in rats. In conclusion, SWRXC can improve the morphology and structure of RA joint tissues, reduce serum factor levels, and may play a role in improving RA by modulating related metabolic pathways such as tryptophan metabolism, nucleotide metabolism and purine metabolism, and altering the composition of intestinal flora. Animal protocols were approved by the Animal Ethics Committee of Qinghai Normal University (No. 2021041203).
6.Single-cell RNA sequencing deciphers transcriptional profiles of hepatocytes in mouse with hepatic alveolar echinococcosis
Qingqing YANG ; Wanzhong JIA ; Xiangqian WANG ; Qigang CAI ; Xin GE ; Wei WANG ; Xiumin HAN
Chinese Journal of Schistosomiasis Control 2023;35(3):236-243
Objective To investigate the cell composition and the transcriptional characteristics in microenvironments of hepatic tissues in mice at late stage of Echinococcus multilocularis infection at a single-cell level. Methods Peri-lesion and paired distal hepatic specimens were collected from two BALB/c mice (6 to 8 weeks old) infected with E. multilocularis for single-cell RNA sequencing. The Seurat package in the R software was employed for quality control of data, multi-sample integration and correction of batch effects, and uniform manifold approximation and projection (UMAP) algorithm was used for cell clustering. Cell types were annotated using classical marker genes. Differentially expressed genes were screened in each cell type through differential gene expression analysis, and the biological roles of cells were predicted using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Results A total of 43 710 cells from peri-lesion and distal hepatic tissues of E. multilocularis-infected mice were analyzed, and were classified into 11 cell types, including neutrophils, T cells, macrophages, granulocyte-monocyte progenitor cells, B cells, plasma cells, basophils, hepatic stellate cells, endothelial cells, hepatocytes, and platelets. T cells were the largest population of immune cells in the microenvironment of hepatic tissues, including five CD4+ T cell subsets, two CD8+ T cell subsets and phosphoantigen-reactive γδT cells. The proportions of CD4+ helper T cells and cytotoxic CD4+ T cells decreased and the proportion of T helper 2 (Th2) cells increased in peri-lesion tissues relative to distal hepatic tissues. In addition, the differentially expressed genes in Th2 cells were associated with negative regulation of the immune system, and the highly expressed genes in cytotoxic CD4+ T cells correlated with activation of the immune system. Conclusions Single-cell RNA sequencing deciphers the cell composition and distribution in microenvironments of hepatic tissues from mice infected with E. multilocularis, and the increased proportion of Th2 cells in peri-lesion hepatic tissues may be associated with formation of immunosuppressive microenvironments.
7.Expression of Key Enzymes in Glucose Metabolism in Chronic Mountain Sickness and Its Correlation with Phenotype.
Yun-Mei GAO ; Guo-Xiong HAN ; Cheng-Hui XUE ; Lai-Fu FANG ; Wen-Qian LI ; Kuo SHEN ; You-Bang XIE
Journal of Experimental Hematology 2023;31(1):197-202
OBJECTIVE:
To explore the pathogenesis of erythrocytosis by detecting the key enzymes of glucose metabolism and glucose transporter in bone marrow erythrocytes of chronic mountain sickness (CMS), and analyzing its correlation with hemoglobin.
METHODS:
Twenty CMS patients hospitalized in Qinghai Provincial People's Hospital from January 2019 to December 2020 were selected as CMS group. Twenty males with leukocyte count > 3.5×109/L who had accepted bone marrow aspiration and had normal result were taken as control group. The mRNA and protein expression of key enzymes and glucose transporter in glucose metabolism in bone marrow CD71+ erythrocytes were detected by real time qPCR and Western blot, respectively. Glucose, lactic acid and 2,3-diphosphoglycerate in the bone marrow supernatant and serum were tested by ELISA. The mRNA and protein expression of key enzymes and glucose transporter, glucose, lactic acid and 2,3-diphosphoglycerate of the two groups were compared. Pearson correlation was used to analyze the correlation between key enzymes, glucose transporter in glucose metabolism in bone marrow CD71+ erythrocytes and hemoglobin.
RESULTS:
The expression of HK2, GLUT1 and GLUT2 mRNA in the CMS group were higher than those in the control group (P<0.001), while the expression of HK1, OGDH and COX5B mRNA were not different. The expression of HK2, GLUT1 and GLUT2 protein in the CMS group were higher than those in the control group (P<0.05). The levels of glucose and lactic acid in the bone marrow supernatant and serum in the CMS group were not different from those in the control group, while the level of 2,3-diphosphoglycerate was higher (P<0.001). Both HK2 and GLUT2 proteins were positively correlated with hemoglobin (r=0.511, 0.717).
CONCLUSION
CMS patients may increase glycolysis by increasing the expression of HK2, and promote the utilization of glucose through high expression of GLUT1 and GLUT2 to meet the need of energy supply.
Male
;
Humans
;
Altitude Sickness/metabolism*
;
Glucose Transporter Type 1
;
2,3-Diphosphoglycerate
;
Hemoglobins
;
Chronic Disease
;
RNA, Messenger
;
Phenotype
;
Glucose
8.Tibial transverse bone transport assisted by nose ring drainage for treatment of foot and ankle chronic osteomyelitis
Huaqing YANG ; Yaohua ZHANG ; Hongyue ZHANG ; Qichang YANG ; Qinghai HAN ; Yun YANG ; Dianzhao ZHAO ; Long QU
Chinese Journal of Orthopaedic Trauma 2023;25(4):302-309
Objective:To investigate the clinical effects of tibial transverse bone transport assisted by nose ring drainage (NRD) in the treatment of foot and ankle chronic osteomyelitis.Methods:A retrospective study was conducted to analyze the data of 32 patients with foot and ankle chronic osteomyelitis who had been treated at Department of Orthopedics, Beijing Rehabilitation Hospital from March 2013 to February 2022. The patients were assigned into a study group and a control group. In the control group, there were 12 males and 3 females, aged (39.5±8.8) years. The osteomyelitis was located at the distal tibia in 4 cases, at the talus in 1 case, at the calcaneus in 2 cases, at the midfoot in 4 cases and at the forefoot in 4 cases. According to the Crerny-Mader classification, there were 5 cases of type Ⅰ and 10 cases of type Ⅱ. The control group was treated with Ilizarov transverse tibial bone transport in combination with thorough debridement and anti-infection therapy. In the study group, there were 12 males and 5 females, aged (42.3±13.4) years. The osteomyelitis was located at the distal tibia in 4 cases, at the talus in 1 case, at the calcaneus in 3 cases, at the midfoot in 5 cases and at the forefoot in 4 cases. According to the Crerny-Mader classification, there were 7 cases of type Ⅰ and 10 cases of type Ⅱ. The study group was treated with NRD drainage in addition to the procedures in the control group. The curative effects were evaluated by comparing the cure, recurrence, amputation (toe amputation), antibiotic use time, wound healing time, relevant inflammatory indicators [WBC, neutrophil percentage (NEU), erythrocyte sedimentation rate (ESR), procalcitonin, interleukin-6 (IL-6), C-reactive protein (CRP)], and postoperative functional recovery between the 2 groups.Results:The 2 groups were comparable because there was no statistically significant difference in the general data between them ( P>0.05). The follow-up period was (36.0±9.8) months. There was no significant difference between the 2 groups in the cure or amputation (toe amputation) ( P>0.05), but the recurrence rate in the study group was significantly lower than that in the control group ( P<0.05). The antibiotic use time [(20.7±3.6) d] and wound healing time [(88.3±17.1) d] in the study group were significantly shorter than those [(37.9±6.5) d and (102.2±22.6) d] in the control group ( P<0.05). The ESR, IL-6 and CRP at 1 week after operation in the study group were significantly lower than those in the control group ( P<0.05), but there was no significant difference between the 2 groups in the WBC, NEU or PCT at 1 week after operation ( P>0.05). There was no significant difference either in the inflammatory indicators between the 2 groups at 1 month after operation ( P>0.05). In all patients, the inflammatory indicators like WBC, NEU, ESR, PCT, IL-6 and CRP at 1 week and 1 month after operation were significantly better than those before operation, and the Maryland foot functional score at 12 months after operation was significantly higher than the preoperative one ( P<0.05). There was no significant difference between the 2 groups in the Maryland foot functional score at 12 months after operation ( P>0.05). Conclusion:In the treatment of foot and ankle chronic osteomyelitis, compared with the Ilizarov transverse tibial bone transport, our tibial transverse bone transport assisted by NRD can achieve satisfactory therapeutic effects, shorten antibiotic use time and treatment cycle, and reduce recurrence rate.
9.Research progress on effect and mechanism of diesel exhaust particles on asthma
Chaohui MU ; Qinghai LI ; Xinjuan YU ; Yize YIN ; Wei HAN
Journal of Environmental and Occupational Medicine 2023;40(5):524-528
As a source of traffic-related air pollution, diesel particulate matter (DPM) associate with a variety of lung-related diseases, but there is no systematic review of the relationship between DPM and the development and progression of asthma. This article reviewed the relationship between DPM and asthma, the effect and mechanism of DPM on airway inflammation and remodeling in asthma, and illustrated that DPM exposure may participate in airway inflammation and remodeling through oxidative stress, immune regulation and regulation of lung and intestinal microecology, so as to promote the development and progression of asthma.
10.Improved ability of demonstrating ocular masses on 3.0 T MR scanner combined with an 8-channel eye surface phased array coil: a multi-center study
Shijun WANG ; Hong JIANG ; Feifei WANG ; Meiyun WANG ; Guanghui BAI ; Qinghe HAN ; Bocheng WANG ; Jingliang CHENG ; Chuanliang CHEN ; Huaizhi GE ; Qinghai YUAN ; Xiaofeng TAO ; Junfang XIAN
Chinese Journal of Radiology 2023;57(1):41-47
Objective:To investigate the value of the 8-channel eye surface phased array coil in improving image quality and demonstrating ocular masses on 3.0 T MR scanner.Methods:From July 2018 to January 2020, the data of orbital MRI in 692 patients with ocular masses on 6 medical centers were prospectively collected. The patients were simple randomly assigned into 8-channel eye surface phased array coil group (413 patients) or 8-channel head phased array coil group (279 patients), with the same MRI sequences. The signal to noise ratio (SNR) and contrast to noise ratio (CNR) were calculated in orbital anatomy structures and masses (eyelid mass, intraocular mass, lacrimal mass and orbital mass). The image quality scores including motion artifact, mass margin, the relationship between the mass and adjacent structures, and overall image quality were recorded. The differences of image quality between the two groups were compared by two independent sample t-test or Wilcoxon rank test. Results:The SNR and CNR were higher in eye surface coil group than those in head coil group ( P<0.05). The scores of ocular movement artifacts were higher in head coil group than those in surface coil group ( P<0.05). The scores of intraocular mass margin, the relationship between the mass and adjacent structures, and overall image quality were higher in surface coil group than those in head coil group ( P<0.001). There were no significant differences in mass margin, the relationship between the mass and adjacent structures, and overall image quality scores of eyelid, lacrimal gland, and orbital mass between the two groups ( P>0.05). Conclusion:3.0 T MR scanner combined with the 8-channel eye surface phased array coil can improve the SNR and CNR of orbital MR images, the demonstration of the intraocular mass margin and the relationship between the mass and adjacent structures.


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