1.Tauroursodeoxycholic acid treats spinal cord injury by reducing apoptosis of spinal cord neurons under glucose and oxygen deprivation
Zepeng CHEN ; Yonghui HOU ; Shudong CHEN ; Yu HOU ; Dingkun LIN
Chinese Journal of Tissue Engineering Research 2024;28(4):528-534
BACKGROUND:Tauroursodeoxycholic acid is a hydrophilic bile acid derivative that has neuroprotective effects in a variety of neurological disease models.However,there are few reports on the effects of tauroursodeoxycholic acid on spinal cord injury. OBJECTIVE:To investigate the effect of tauroursodeoxycholic acid on apoptosis of spinal cord neurons under hypoglycemic and hypoxic conditions,as well as the effect on recovery of motor function in mice after spinal cord injury. METHODS:(1)In vitro experiment:Primary spinal cord neurons were isolated from C57 BL/6 mouse embryos at 13.5 days of gestation.After 72 hours of culture,the cells were divided into three groups.In the normal group,cells were cultured in Neurobasal complete medium that was incubated in a CO2 incubator(5%CO2 + 95%air)for 24 hours.In the oxyglucose-deprived group,sugar-free Neurobasal medium was added and incubated in a triple-gas incubator(94%N2+5%CO2+1%O2)for 12 hours,and then the medium was replaced with Neurobasal complete medium and incubated in a CO2 incubator for 12 hours.In the experimental group,the treatment procedure was approximately the same as that in the oxyglucose-deprived group,except that taurodeoxycholic acid was added along with the sugar-free Neurobasal medium.TUNEL staining was used to detect apoptosis,cell counting kit-8 assay was applied to detect cell activity,and immunofluorescence staining was performed to detect cellular β-microtubule protein expression.(2)Animal experiment:Sixty C57 BL/6 mice were randomly divided into sham-operated group,spinal cord injury group and experimental group,with 20 mice in each group.Animal models of T9-T10 spinal cord injury were established using Allen's percussion method in the spinal cord injury group and the experimental group.Starting from the 1st day after modeling,taurodeoxycholic acid solution was given by gavage in the experimental group and normal saline was given by gavage in the sham-operated and spinal cord injury groups once a day for 14 consecutive days.Spinal cord tissue repair was assessed using behavioral and histological methods. RESULTS AND CONCLUSION:In vitro experiment:TUNEL staining,cell counting kit-8 and immunofluorescence staining showed that compared with the normal group,the number of apoptotic cells was higher(P<0.01),while cell activity and β-microtubule protein expression were lower in the oxyglucose-deprived group(P<0.01);compared with the oxyglucose-deprived group,the number of apoptotic cells was lower(P<0.01),while cell activity and β-microtubule protein expression were higher in the experimental group(P<0.01).Animal experiment:The Basso-Beattie-Bresnahan scores in the open field test and hind limb footprint experiments showed that the mice in the experimental group had better recovery of walking and motor functions than those in the spinal cord injury group.Hematoxylin-eosin staining showed that significant deformities and cavities were observed at the site of spinal cord injury and the number of nerve cells was significantly reduced in the spinal cord injury group.Compared with the spinal cord injury group,the experimental group showed a significant reduction in the area of spinal cord injury,less spinal cord deformity,fewer cavities,and an increase in the number of nerve cells.Immunofluorescence staining showed that the number of neuronal nucleus-labeled neuronal cells in the spinal cord injury group was less than that in the sham-operated group(P<0.01),and the number of neuronal nucleus-labeled neuronal cells in the experimental group was higher than that in the spinal cord injury group(P<0.01).To conclude,tauroursodeoxycholic acid could effectively reduce glucose/oxygen deprivation-induced apoptosis of spinal cord neurons and axonal loss,and promote the recovery of motor function in mice with spinal cord injury.
2.Analysis of the types and functions of CD34 + cells in full-thickness skin defect wounds of normal mice and diabetic mice by single-cell RNA sequencing
Jia HE ; Jingru WANG ; Wenjun GAN ; Guiqiang LI ; Qi XIN ; Zepeng LIN ; Shubin RUAN ; Xiaodong CHEN
Chinese Journal of Burns 2024;40(3):230-239
Objective:To analyze the types and functions of CD34 + cells in full-thickness skin defect wounds of normal mice and diabetic mice by single-cell RNA sequencing. Methods:This study was an experimental study. The CD34 + cell lineage tracing mouse was produced, and the visualization of CD34 + cells under the fluorescent condition was realized. Six male CD34 + cell lineage tracing mice aged 7-8 weeks (designated as diabetic group) were intraperitoneally injected with streptozotocin to establish a diabetic model, and full-thickness skin defect wounds were prepared on their backs when they reached 13 weeks old. Another 6 male CD34 + cell lineage tracing mice aged 13 weeks (designated as control group) were also subjected to full-thickness skin defect wounds on their backs. On post-injury day (PID) 4, wound tissue was collected from 3 mice in control group and 2 mice in diabetic group, and digested to prepare single-cell suspensions. CD34 + cells were screened using fluorescence-activated cell sorting, followed by single-cell RNA sequencing. The Seurat 4.0.2 program in the R programming language was utilized for dimensionality reduction, visualization, and cell clustering analysis of CD34 + cell types, and to screen and annotate the marker genes for each CD34 + cell subpopulation. Kyoto encyclopedia of genes and genomes (KEGG) and gene ontology (GO) enrichment analysis was performed to analyze the differentially expressed genes (DEGs) of CD34 + fibroblasts (Fbs), smooth muscle cells (SMCs), keratinocytes (KCs), and chondrocyte-like cells (CLCs) in the wound tissue of two groups of mice for exploring cellular functions. Results:On PID 4, CD34 + cells in the wound tissue of both groups of mice were consisted of 7 cell types, specifically endothelial cells, Fbs, KCs, macrophages, T cells, SMCs, and CLCs. Among these, Fbs were further classified into 5 subpopulations. Compared with those in control group, the proportions of CD34 + endothelial cells, Fbs subpopulation 1, Fbs subpopulation 4, KCs, and CLCs in the wound tissue of mice were increased in diabetic group, while the proportions of CD34 + Fbs subpopulation 2, Fbs subpopulation 3, and SMCs were decreased. The marker genes for annotating CD34 + CLCs, endothelial cells, Fbs subpopulation 1, Fbs subpopulation 2, Fbs subpopulation 3, Fbs subpopulation 4, Fbs subpopulation 5, KCs, macrophages, SMCs, and T cells were respectively metastasis-associated lung adenocarcinoma transcript 1, fatty acid binding protein 4, Gremlin 1, complement component 4B, H19 imprinted maternally expressed transcript, Dickkopf Wnt signaling pathway inhibitor 2, fibromodulin, keratin 5, CD74 molecule, regulator of G protein signaling 5, and inducible T-cell co-stimulator molecule. KEGG and GO enrichment analysis revealed that, compared with those in control group, DEGs with significant differential expression (SDE) in CD34 + Fbs from the wound tissue of mice in diabetic group on PID 4 were significantly enriched in terms related to inflammatory response, extracellular matrix (ECM) organization, regulation of cell proliferation, and aging (with Pvalues all <0.05), DEGs with SDE in CD34 + SMCs were significantly enriched in terms related to cell migration, apoptotic process, positive regulation of transcription, and phagosome (with P values all <0.05), DEGs with SDE in CD34 + KCs were significantly enriched in terms related to mitochondrial function, transcription, and neurodegenerative diseases (with P values all <0.05), and DEGs with SDE in CD34 + CLCs were significantly enriched in terms related to rhythm regulation, ECM, and viral infection (with P values all <0.05). Conclusions:CD34 + cells display high heterogeneity in the healing process of full-thickness skin defect wounds in both normal mice and diabetic mice. The significantly enriched functions of DEGs with SDE in CD34 + cell subpopulations in the wound tissue of the two mouse groups are closely related to the wound healing process.
3.The role of effective albumin in chronic liver disease
Jun LIN ; Zepeng LI ; Junying YU
Basic & Clinical Medicine 2024;44(12):1707-1711
Effective albumin refers to serum albumin with intact structure and function.In the course of chronic liver disease,some serum albumin loses its functional integrity through different post-translational modifications such as oxidation and glycosylation.Level of effective albumin is lower in patients with decompensated cirrhosis and is significantly correlated with patient survival,underscoring that they are more sensitive than total serum al-bumin in reflecting the actual state of liver function.In addition,effective albumin not only serves as a predictor,but also plays a key role in the treatment of chronic liver disease by improving albumin preparations and develo-ping new therapeutics.
4.Research on mental health status of medical staff during COVID-19 epidemic
Zepeng HUANG ; Kaiting ZHANG ; Xiaoyi HE ; Wanbao YE ; Zeting LIN ; Limin OU ; Liping LI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2020;38(11):834-838
Objective:To explore the psychological status of medical staff during COVID-19 epidemic, so as to provide reference and scientific basis for carrying out further psychological intervention and ensuring the mental health of medical staff.Methods:By using convenient sampling method and the Stresss-Anxiety ubscale of Depression Anxiety Stress (DASS-21) , the mental health status of 615 medical staff was investigated by the way of questionnaire star from February 4 to 16, 2020. A total of 615 questionnaires were distributed and collected, and 615 were valid, with an effective recovery rate of 100%.Results:The detection rates of psychological stress and anxiety of medical staff were 13.82% (85/615) and 25.37% (156/615) , respectively. 31-40 years old and working in key departments were risk factors for psychological stress of medical staff ( OR=1.779, 2.127) ; Women, frequently washing hands with soap/hand sanitizer/disinfectant were protective factors for psychological stress ( OR=0.520, 0.528) . Medical staff working in designated hospitals and key departments were more likely to have anxiety ( OR=2.042, 2.702) ; The high fit of the mask to the face and bridge of the nose was a protective factor for the psychological anxiety of medical staff ( OR=0.500) . Conclusion:Medical staff show higher stress and anxiety during the epidemic of COVID-19. Psychological intervention should be carried out early, focusing on men, age 31 to 40, medical staff working in designated hospitals and key departments.
5.Research on mental health status of medical staff during COVID-19 epidemic
Zepeng HUANG ; Kaiting ZHANG ; Xiaoyi HE ; Wanbao YE ; Zeting LIN ; Limin OU ; Liping LI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2020;38(11):834-838
Objective:To explore the psychological status of medical staff during COVID-19 epidemic, so as to provide reference and scientific basis for carrying out further psychological intervention and ensuring the mental health of medical staff.Methods:By using convenient sampling method and the Stresss-Anxiety ubscale of Depression Anxiety Stress (DASS-21) , the mental health status of 615 medical staff was investigated by the way of questionnaire star from February 4 to 16, 2020. A total of 615 questionnaires were distributed and collected, and 615 were valid, with an effective recovery rate of 100%.Results:The detection rates of psychological stress and anxiety of medical staff were 13.82% (85/615) and 25.37% (156/615) , respectively. 31-40 years old and working in key departments were risk factors for psychological stress of medical staff ( OR=1.779, 2.127) ; Women, frequently washing hands with soap/hand sanitizer/disinfectant were protective factors for psychological stress ( OR=0.520, 0.528) . Medical staff working in designated hospitals and key departments were more likely to have anxiety ( OR=2.042, 2.702) ; The high fit of the mask to the face and bridge of the nose was a protective factor for the psychological anxiety of medical staff ( OR=0.500) . Conclusion:Medical staff show higher stress and anxiety during the epidemic of COVID-19. Psychological intervention should be carried out early, focusing on men, age 31 to 40, medical staff working in designated hospitals and key departments.
6.Osteogenesis of dextran sulfate/recombinant human bone morphogenetic protein-2/chitosan nano microspheres combined with coralline hydroxyapatite in repair of large segmental bone defects
Zepeng CHEN ; Yuanjun XIA ; Leng HAN ; Xiangling YE ; Zefeng LIN ; Ying ZHANG
Chinese Journal of Orthopaedic Trauma 2017;19(12):1074-1080
Objective To investigate the osteogenic ability of dextran sulfate/recombinant human bone morphogenetic protein-2/chitosan (DS/rhBMP2/CS) nano microspheres combined with coralline hydroxyapatite (CHA) in repair of segmental bone defects.Methods DS/rhBMP2/CS microspheres prepared by ionic crosslinking method were adsorbed into CHA by lyophilization.Seventy-two New Zealand rabbits were randomly divided into 3 equal groups after they had been made into models of bone defect at the right radius.The defects in the 3 groups were implanted respectively with CHA,rhBMP2/CHA and DS/rhBMP2/ CS/CHA.Another 18 animals served as a blank control group.Blood and bone samples were obtained at 4,8 and 12 weeks after implantation.The serum BGP was detected,and the bone grafts were scanned by micro-CT for calculation of volume ratio of the new bone.Hematoxylin and eosin (HE) staining was performed after bone decalcification.Results All the 72 animals recovered well without any infection or graft exposure.Gross observation at postoperative 12 weeks showed that the DS/rhBMP2/CS/CHA group was the best in the quality,quantity and strength of the new bone,as well as in the healing of bone defects.The serum levels of bone gamma-carboxyglutamic-acid-containing protein at all time points in the DS/rhBMP2/CS/CHA group were significantly higher than those in the CHA and rhBMP2/CHA groups (P < 0.05).Micro-CT scanning demonstrated obvious progress in bone formation,cortical bone and marrow cavity at all time points in the DS/rhBMP2/CS/CHA group which showed significantly faster bone reconstruction synchronized with material degradation and significantly higher volume ratio of the new bone than the other 2 groups (P < 0.05).Histological examinations showed better morphology of mature cortical bone and new marrow cavity at all time points in the DS/rhBMP2/CS/CHA group than in the other 2 groups.Conclusion Since DS/rhBMP2/CS/CHA possesses a better mechanism of sustained-releasing rhBMP2 to induce bone formation because of its reticular and hole-hole-connected structure,it may perform better in repairing segmental bone defects than simple CHA or rhBMP2/CHA.
7.Therapeutic effect of olmesartan on essential hypertension and its influence on cardiovascular risk fac-tors
Zepeng LIN ; Xiaohong SUN ; Zhiwei ZHANG
Chinese Journal of cardiovascular Rehabilitation Medicine 2015;24(3):310-312
Objective:To evaluate therapeutic effect of olmesartan on patients with mild to moderate essential hyper-tension (EH) and its influence on cardiovascular risk factors .Methods:A total of 150 mild/moderate EH patients were enrolled using opened ,non-control and multi-center method .According to random number table ,they were randomly and equally divided into olmesartan group (received olmesartan 20~40 mg/d) and losartan group (re-ceived losartan 50~100 mg/d) .Changes of systolic blood pressure (SBP) ,diastolic blood pressure (DBP) , levels of risk indexes of cardiovascular diseases ,such as high sensitive C reactive protein (hsCRP) ,urinary albumin/creati-nine ,homocysteine (Hcy) and carotid intima-media thickness (IMT) were observed in two groups before and after treatment .Results:After eight-month treatment , compared with before treatment , all indexes significantly im-proved in both groups;compared with losartan group ,there were significant reductions in SBP [ (145 ± 12) mmHg vs .(132 ± 9) mmHg] and DBP [ (90 ± 7) mmHg vs .(80 ± 5) mmHg] , P<0.01 all;and significant reductions in levels of hsCRP [ (0.54 ± 0.09) mg/L vs .(0.34 ± 0.09) mg/L] ,urinary albumin/creatinine [ (26.5 ± 19.4) mg/g vs .(23.1 ± 18.3) mg/g] ,Hcy [ (13.2 ± 4.8)μmol/L vs .(10.3 ± 4.2)μmol/L] and IMT [ (0.83 ± 0.29) mm vs . (0.74 ± 0.28) mm] in olmesartan group , P< 0.05 all .Conclusion:Besides intensively lowering blood pressure olmesartan can also significantly decrease cardiovascular risk factors in patients with mild to moderate hypertension .
8.Changes of serum concentration of amino-terminal propeptide of procollagen type Ⅲ(P Ⅲ NP)of post-burn hypertrophic scar at excessive stages
Rui SHEN ; Xiangsheng FENG ; Hongmei SONG ; Fenggang ZHANG ; Xiaodong CHEN ; Shubin RUAN ; Zepeng LIN ; Yan LIN ; Jingqun ZHANG
International Journal of Surgery 2010;37(11):726-728
Objective To explore the level and the significance of amino-terminal propeptide of procollagen type Ⅲ(P Ⅲ NP)in the serum of hypertrophic scar patients at excessive stages.Methods The serum samples of 74 inpatients admmitted in Affiliated Fosham Hospital of Sun Yat-sen University from August 2007 to August 2009 suffering from long-persisting post-burn hypertrophic scar at various stages were collected.Serum samples of 12 were normal persons were also collected and used as control group.Hypertrophic scar group were divided into 4 subgroups according to the phase of scar and the serum concentrations of P Ⅲ NP were detected using the sensitive RIA method.Results The level of PⅢ NP increases gradually during the process of immature hypertrophic scar to mature scar before it peaking in 4 to 6 months scar subgroup.The concentration of PⅢ NP degreased gradually with the maturation of hypertrophic scar.Conclusions The high concentration of PⅢNP can reflect the high level of the synthesis of type Ⅲ collagen in the tissue of post-burn hypertrophic scar.The level of P Ⅲ NP is synonymous with the ongoing process of hypertrophic scar.PⅢ NP may be a satisfactory marker in discerning the growth and development of post-burn hypertrophic scar.
9.Blood Glucose Levels and Severity of Coronary Stenosis in Hypertensive Patients
Xiaoling PENG ; Zepeng LIN ; Zhiwei ZHANG ; Guanglin JIN ; Changnong PENG ; Wen WANG
Chinese Journal of Hypertension 2006;0(12):-
Objective To investigate the relationship between blood glucose levels and severity of coronary stenosis in hypertensive patients. Methods Blood pressure, fasting plasma glucose, 2 h plasma glucose, clinical features and coronary angiographic findings were analyzed retrospectively in 540 patients with essential hypertension. Acoording 2 h plasma glucose, patients were stratified into three group: group 1: 2h plasma glucose
10.Effect of Rosuvastatin on Microalbuminuria and Arteriosclerosis Plaque of Artery in Hypertensive Patients
Xiaoling PENG ; Zepeng LIN ; Hong TENG ; Rongkui ZHANG ; Zhiwei ZHANG
Chinese Journal of Hypertension 2006;0(08):-
Background Rosuvastatin has been unanimously recognized as an very highly efficacious statin. Rosuvastatin significantly reduced microalbuminuria and its powerful anti-inflammatory effect confer anti-atherosclerosis in patients with essential hypertension. Objective To study the influence of rosuvastatin on microalbuminuria and arteriosclerosis plaque of carotid and coronary arteries in hypertensive patients. Methods Seventy-six hypertensive patients were randomly to receive conventional anti-hypertensive drugs (amlodipine 5 mg/d and telmisartan 80 mg/d,n=37) or rosuvastatin (10 mg QN) on the top of conventional anti-hypertensive drugs (n=39). The blood pressure,levels of microalbuminuria,echocardiography and helical computerized tomography were examined before and 8 months after treatment in all patients. Results After 8 months of treatment,SBP and DBP were decreased in both group [conventional treatment from (166.3?11.2)/(92.4?8.2) to (133.6?9.8)/(85.5?6.1)mmHg,combined group from (165.6?10.5)/(91.5?6.7) to (128.1?9.2)/(81.1?5.9)mmHg]. Combined treatment significantly reduced the levels of microalbuminuria [(31.6?21.8) to (23.2?19.8)mg/g,P

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