1.Exploration and clinical validation of the repair mode of the sclerotic zone of steroid-induced osteonecrosis of the femoral head based on Tandem Mass Tags technology
Zhikun ZHUANG ; Mincong HE ; Tianye LIN ; Rongkai WU ; Jinhua GUO ; Zhaoke WU ; Qiushi WEI
Chinese Journal of Tissue Engineering Research 2024;28(14):2191-2196
BACKGROUND:The sclerotic zone in the femoral head is an important imaging feature in the progression of steroid-induced femoral head necrosis,which is associated with disease prognosis.Peroxisome proliferator-activated receptor γ coactivator 1α(PGC-1α)has been shown to possess biological activities such as osteogenesis,angiogenesis and anti-mitochondrial apoptosis,which may be closely related to bone repair of steroid-induced femoral head necrosis. OBJECTIVE:To screen for the differential proteins in the sclerotic zone of steroid-induced osteonecrosis of the femoral head versus the normal zone,to screen for hub proteins in the sclerotic zone,and to verify the differential expression of hub proteins in the femoral head specimens following steroid-induced femoral head necrosis,and to to explore the repair pattern of the sclerotic zone following steroid-induced femoral head necrosis. METHODS:Femoral head samples were collected from patients with steroid-induced osteonecrosis of the femoral head receiving total hip arthroplasty.The differentially expressed genes in the sclerotic zone and the normal zone were screened by Tandem Mass Tags and analyzed by GO and KEGG signaling pathways to construct a protein-protein interaction network and screen hub genes.In addition,the expression of hub genes in the sclerotic zone was verified by immunohistochemistry and western blot. RESULTS AND CONCLUSION:Quantitative protein profiling by Tandem Mass Tags revealed that 609 proteins were significantly differentially expressed(Log2FC>1.20,Log2FC<0.84 and P<0.05)in the sclerotic zone of the femoral head compared with the normal zone,of which 290 proteins were upregulated and 319 proteins were downregulated.The GO and KEGG pathway enrichment analyses revealed that among the top 10 enriched pathways,Wnt signaling pathway and life-cycle regulatory pathway were closely related to bone repair;in the life-cycle regulatory pathway,PGC-1α was one of the important proteins.In addition,western blot results verified the low expression of PGC-1α and NRF1 in the sclerotic zone and high expression of Cleaved Caspase-3 in the sclerotic zone compared with the normal zone of steroid-induced femoral head necrosis specimens.Light microscopic immunohistochemical results showed the distribution of PGC-1α,NRF1 and Cleaved Caspase-3 positive expression in the sclerotic and normal zones in the femoral head tissue specimens,indicating the presence of their expression in bone trabeculae,osteoblasts and bone marrow.In contrast,the brown area of the sclerotic zone of femoral head necrosis stained darker and showed more obvious expression of Cleaved Caspase-3.To conclude,in the sclerotic zone of steroid-induced femoral head necrosis,biological behaviors including activation of osteogenesis-related pathways such as Wnt and oxidative apoptosis characterized by low expression of PGC-1 are observed.Low expression of PGC-1α in the sclerotic zone of steroid-induced femoral head necrosis may be associated with the activation of oxidative apoptosis.
2.Establishment of HPLC fingerprint of Yi drugs of MaRSDenia tenacissima and evaluation of chemometrics
Zhikun ZHU ; Yuanyuan GUO ; Piao WANG ; Wubing WANG ; Siqi LI ; Lingyun CHEN
International Journal of Traditional Chinese Medicine 2023;45(12):1549-1554
Objective:To establish the HPLC fingerprint of MaRSDenia tenacissima; To evaluate the different origins of MaRSDenia tenacissima by combining chemometric methods.Methods:High performance liquid chromatography (HPLC) method was adopted, with DiKMA C18 column (250 mm× 4.6 mm, 5 μm), acetonitrile-0.05% phosphoric acid aqueous solution as mobile phase gradient elution, flow rate of 1.0 ml/min. The detection wavelength was set at 230 nm, the column temperature was 30 ℃, and sample size was 10 μl. Chromatographic information was imported into the similarity evaluation system for TCM chromatographic fingerprints (2012 version) for similarity analysis. SPSS Statistics 26 was used for system clustering analysis, and SIMCA 14.1 software was used for principal component analysis and partial least squares discriminant analysis (PLS-DA).Results:Totally 12 common peaks were identified. Two chromatographic peaks were identified as tenacissoside G and tenacissoside I. The relative similarity of fingerprints of 15 batches of samples and references ranged from 0.942 to 0.995. When the square Euclidean distance was 20, the samples could be grouped into two categories: S1-S3, S13-S15 were grouped into one category, and S4-S12 were grouped into another category. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) showed that there were significant differences among 15 batches of MaRSDenia tenacissima, and there was a certain correlation with the origin.Conclusion:The results can provide a reference for analyzing the differences of MaRSDenia tenacissima from different producing areas and the quality standards of related formula granules in the later stage.
3.The predictive value of combined detection of serum ANGPTL8 and VCAM-1 levels for cerebral vasospasm after intracranial aneurysm embolization
Mengmeng WEI ; Zhikun LV ; Guozhen LI ; Xueyan HU ; Qianqian XU ; Jing GUO ; Biao GENG
Journal of Clinical Surgery 2023;31(12):1133-1136
Objective The combined detection of serum angiopoietin-like protein 8(ANGPTL8)and Vascular cell adhesion molecule-1(VCAM-1)levels was analyzed for the predictive value of cerebral vasospasm(CVS)after intracranial aneurysm embolization.Methods A total of 196 patients who underwent intracranial aneurysm embolization in our hospital from March 2019-March 2022 were selected as the study subjects,99 patients with CVS were in the CVS group,and 97 patients without CVS were in the non CVS group.Serum ANGPTL8 and VCAM-1 levels were detected by ELISA;the correlation between serum ANGPTL8 and VCAM-1 levels was analyzed by Pearson method,Logistic regression was used to analyze the influencing factors of CVS in patients undergoing intracranial aneurysm embolization;ROC curve was used to analyze the serum levels of ANGPTL8 and VCAM-1 to predict the cutoff value of CVS in patients undergoing intracranial aneurysm embolization;four grid table method was used to analyze the predictive value of ANGPTL8,VCAM-1 and their combination on the occurrence of CVS in patients undergoing intracranial aneurysm embolization.Results The differences between CVS and non-CVS groups were statistically significant in hypertension,Hunt-Hess grade,and Glasgow coma(GCS)scores(P<0.05).The serum ANGPTL8 and VCAM-1 levels in the CVS group were significantly higher than those in the non-CVS group(P<0.05).There was a positive correlation between serum ANGPTL8 and VCAM-1(r=0.468,P<0.05).Multivariate analysis showed that high level of ANGPTL8(OR=3.652,95%CI:1.434-9.302),high level of VCAM-1(OR=2.619,95%CI:1.212-5.658),Hunt Hess grade Ⅲ-Ⅳ(OR=1.927,95%CI:1.104-3.362),GCS score of 3-8(OR=2.813,95%CI:1.257-6.295)were independent risk factors for CVS in patients undergoing intracranial aneurysm embolization.The AUC of serum ANGPTL8 level in predicting CVS in patients undergoing intracranial aneurysm embolization was 0.844,and the cut-off value was 189.233 U/L;the AUC of serum VCAM-1 level in predicting CVS in patients undergoing intracranial aneurysm embolization was 0.795,and the cutoff value was 17.984 mg/L.The accuracy,sensitivity and specificity of the combined prediction for CVS were 89.81%,93.94%and 85.57%,respectively,which were obviously higher than those of the single prediction.Conclusion The serum levels of ANGPTL8 and VCAM-1 in CVS group are obviously higher than those in non CVS group.The combination of the two has a high predictive value for CVS after intracranial aneurysm embolization.
4.Analysis of risk factors in early infection patients after heart transplantation
Yunfei LIU ; Xiangli ZHANG ; Zhiying LI ; Zhikun FU ; Keke LIANG ; Lin GUO
Chinese Journal of Organ Transplantation 2022;43(7):406-411
Objective:To explore the risk factors of early infection patients after heart transplantation(HT)and provide references for preventing and treating early infection.Methods:From April 2018 to May 2021, clinical data were retrospectively reviewed for 95 HT recipients treated at Zhengzhou Seventh People's Hospital.They were divided into two groups of infected(n=34)and uninfected(n=61). Gender, age, disease type, preoperative IABP implantation, postoperative intra-aortic balloon pump(IABP)implantation, postoperative extracorporeal membrane oxygenation(ECMO)implantation, preoperative mechanical ventilation, preoperative leukocyte, preoperative lymphocyte, preoperative serum C-reactive protein(CRP), operative approach, APACHEⅡscore, NYHA grade, hemoglobin, cardiopulmonary bypass time, donor heart cold ischemia time, postoperative thoracic drainage tube indwelling time, postoperative gastric tube indwelling time, postoperative urinary tube indwelling time, postoperative acute rejection, postoperative ventilator assisted treatment time and postoperative ICU time.The risk factors of early infection were analyzed by univariate and multivariate Logistic regression analysis.Results:There were 34 cases of early infection after HT and 8 cases died.In infection group, preoperative hemoglobin(female <110 g/L or male <120 g/L), ECMO post-operation, 24-48 h post-operation, APACHE post-operation(>6), postoperative intrathoracic drainage tube indwelling time(≥7 d), postoperative gastric tube indwelling time(≥4 d), postoperative urinary tube indwelling time(≥5 d), postoperative acute rejection(positive), postoperative ventilator assisted treatment time(≥2 d)and postoperative ICU time(≥10 d)were 18 cases(52.94%), 8(23.53%), 30(88.24%), 22(64.71%), 18(52.94%), 20(58.82%), 4(11.76%), 21(61.76%)and 19(55.88%); uninfected group: 16 cases(26.23%), 3(4.92%), 32(52.46%), 24(39.34%), 15(24.59%), 31(34.43%), 1(1.64%), 21(34.43%)and 4(6.56%). Significant inter-group differences existed( χ2=6.778, 5.68, 12.326, 5.623, 7.740, 5.297, 4.489, 6.615, 28.947, P<0.05). Multivariate Logistic regression analysis indicated that 24-48h post-operation, APACHEⅡ score >6(β=1.024, Wald χ2=7.653, OR=2.141, OR95% CI=1.323~4.215), ECMO post-operation(β=1.783, Wald χ2=6.186, OR=5.949, OR95% CI =1.459~24.25), postoperative intrathoracic drainage tube indwelling time ≥7 d(β=0.712, Wald χ2=5.745, OR=1.054, OR95% CI=1.183~6.753), postoperative gastric tube indwelling time(β=0.832, Wald χ2=6.756, OR=1.132, OR95% CI=1.416~8.406), postoperative ventilator assisted treatment time(β=0.745, Wald χ2=6.563, OR=1.212, OR95% CI=1.289~7.346)and postoperative ICU time=1.28(β=1.325, Wald χ2=9.752, OR=2.435, OR95% CI=1.426~6.354)were independent risk factor for early infection after HT( P<0.05). Conclusions:Early infection after HT remains higher.It is significantly correlated with 24-48 h post-operation APACHE II score, ECMO post-operation, postoperative intrathoracic drainage tube indwelling time, postoperative gastric tube indwelling time, postoperative ventilator assisted treatment time and postoperative ICU time.Targeted interventions should be adopted for lowering the incidence of early infection after HT.
5.Oral anticoagulants for prevention of stroke in patients with atrial fibrillation
Beiping WANG ; Meng CUI ; Zhikun WU ; Xiujuan SONG ; Li GUO
International Journal of Cerebrovascular Diseases 2018;26(6):450-453
Atrial fibrillation (AF) is a common arrhythmia type. It affects more than 80% of the elderly. The main hazards are stroke and other systemic thromboembolism. Compared with the strokes caused by other causes, the disability, mortality and recurrence rate of AF-related stroke are higher. Therefore, the prevention of stroke in patients with AF is critical. Anticoagulation is the core therapeutic strategy for stroke prevention in patients with AF. This article reviews various oral anticoagulants for stroke prevention in patients with AF.
6.Decreased c-kit and miR-21 gene expression in rat heart failure
Zhen LIU ; Shuo LI ; Lingling LIU ; Zhikun GUO ; Pengfei WANG
Basic & Clinical Medicine 2017;37(1):76-79
Objective To explore mechanism ( s) is dynamic expression and internal relations of c-kit gene and miR-21 in rat ventricular remodeling .Methods SD rats( n=140) were randomly divided into normal control group ( n=15 ) and heart failure model group ( n=125 ) .Heart failure model:4 mg/kg intraperitoneal injection of adria-mycin.To detect the cardiac function of rats in eight weeks , proving the heart failure .Then harvest the rat heart , frozen section , using immunohistochemistry and immunofluorescence color to detect the expression changes of miR-21 and c-kit in myocardial tissue .Results The emergence of the pathological changes of cardiac muscle cells after myocardial infarction in the heart failure groups and the control group didn't appear the myocardial infarction and heart failure .Immunohistochemistry and immunofluorescence show that miR-21 positive cells in normal and heart failure myocardium mainly express in vascular endothelium ,a few myocardial cells and stem cells , and endocardial expressing quantity fell more than epicardial;c-kit positive cells tend to cluster together , mainly gather in the epi-cardial and its nearby , and the expressing quantity decrease significantly after heart failure .A small number of cells exsit miR-21 and c-kit common expression in both groups ( P<0.05 ) .Conclusions The decreased expression of c-kit and miR-21 is highly related to rats heart failure and left ventricular remodeling .
7.Effect of warming yang and benefiting marrow recipe on the expression of collagen type II and proteoglycans in the articular cartilage
Lin JING ; Zhikun GUO ; Hongmei ZHANG ; Mingjiang HE ; Pengcheng SHAN
Chinese Journal of Tissue Engineering Research 2015;(24):3798-3802
BACKGROUND:Col agen type II and proteoglycan loss are two most obvious manifestations of cartilage damage in the onset of osteoarthritis. Changes in col agen type II and proteoglycan as the main components of cartilage matrix directly lead to cartilage degeneration and subsequently result in osteoarthritis. How to reverse or prevent the development of this process becomes the focus of medical research. OBJECTIVE:To observe the effect of warming yang and benefiting marrow recipe on the expression of col agen type II and proteoglycan in the articular cartilage of knee osteoarthritis rabbits as wel as to further explore the mechanism underlying chondrocyte protection. METHODS:Ninety-six New Zealand rabbits, aged 9 months old, male and female, were selected to prepare osteoarthritis models in extension position using cast immobilization method, and were randomly divided into four groups:blank group (untreated), model group (simple modeling), Chinese medicine group (intragastric administration of extracts of warming yang and benefiting marrow recipe, 24 mL/kg/d) and western medicine group (intragastric administration of glucosamine hydrochloride, 24 mL/kg/d). Intragastric administration was done once a day for 6 weeks. RT-PCR technology was used to observe the effect of warming yang and benefiting marrow recipe on the expression of col agen type II and proteoglycans in the articular cartilage, and pathological examination was also done. RESULTS AND CONCLUSION:The cartilage surface was smooth in the blank group and Chinese medicine group, with uniform toluidine blue staining, but in the model group and western medicine group, the cartilage surface was rough and the toluidine blue staining was extremely uneven with obvious loss of surface and middle layer dying. The expressions of cartilage proteoglycan and col agen type II in the model group were significantly lower than those in the blank group (P<0.01) as wel as in the Chinese medicine group and western medicine group (P<0.05). In addition, the expressions of cartilage proteoglycan and col agen type II in the Chinese medicine group were higher than those in the western medicine group (P<0.05). These findings indicate that the recipe of warming yang and benefiting marrow can enhance the expressions of col agen type II and proteoglycan, which can maintain the normal col agen phenotype and protect the articular cartilage.
8.A new method to study nerve fiber projecting in the spinal cord during chicken embryo development
Azhen HU ; Ciqing YANG ; Sulei FU ; Yangyang JIA ; Han LI ; Zhikun GUO ; Juntang LIN
Acta Anatomica Sinica 2014;(5):724-728
Objective To develop a method of studying fiber projecting in the spinal cord duiring chicken embryo development.Methods At embryonic incubation 3 day (E3), pCAGGS-green fluorescent protein (GFP) plasmid was injected into the spinal cord using in vivo electroporation.Three days after transfection (E6), GFP-positive embryos were collected under a stereo fluorescence microscope .Subsequently , the spinal cord was separated from the embryos and cut from the roof plate as an open book .After fixed with 4%paraformaldehyde ( PFA) for one hour , the opened spinal cords were used for immunohistochemistry with N-cadherin antibody and with DAPI for nuclei .Finally, the nerve fiber projecting was photographed and analyzed under a fluorescence microscope . Results Based on the opened spinal cord and immunostaining in the cryosection , we observed that the nerve fibers projected across the midline of the floor plate and reached to the sulcus terminalis along the white matter of the contra side .The immunoreaction against N-cadherin indicated that overexpression of GFP has no significant effect on chicken embryonic development .Conclusion A new method to study fiber projecting in the developing chicken spinal cord is established successfully in this study .
9.Clinical value of Hangzhou Criteria in selection of patients with hepatocellular carcinoma for living donor liver transplantation
Shusen ZHENG ; Kai WANG ; Xiao XU ; Qi LING ; Jun CHEN ; Min ZHANG ; Weilin WANG ; Jian WU ; Zhikun LIU ; Haijun GUO
Chinese Journal of Organ Transplantation 2011;32(6):330-333
Objective To investigate the clinical value of Hangzhou Criteria in selection of patients with hepatocellular carcinoma(HCC)for living donor liver transplantation(LDLT)and their effects on recipient's post-transplant survival.Methods A retrospective analysis was conducted on 43 patients with HCC who underwent LDLT in our center.All of the recipients were classified into subgroups according to Milan Criteria,Up-to-Seven Criteria and Hangzhou Criteria,and post-transplant total survival rate and tumor-free survival rate were compared among the criteria.Results The number of patients within Hangzhou Criteria was 61.5 %(8/13)more than that within Milan Criteria and 23.5 %(4/17)more than that within Up-to-Seven Criteria.The 1-,and 3-years total survival rate and tumor-free survival rate of the recipients within Milan Criteria(n=13)were 100 %,80.0 % and 84.6 %,84.6 %,those of the recipients within Up-to-Seven Criteria(n=17)were 100 %,75.2 % and 87.5 %,81.2 %,and those of the recipients within Hangzhou Criteria(n=21)were 100 %,80.0 % and 89.5 %,84.2 %,respectively.There was no significant difference among the three criteria in total survival rate and tumor-free survival rate(P>0.05).Conclusion Hangzhou Criteria further enrolled more liver transplantation candidates without decline of total survival rate and tumor-free survival.It is a more effective selection of HCC recipients for LDLT.
10.Strengthening Information Literacy Education and Innovating Traditional Chinese Medicine Talents Cultivating Mode
Journal of Medical Informatics 2009;30(8):92-95
Based on literature review, questionnaire survey, Interact investigation, experts interview and seminars the paper analyzes the status quo of the information literacy education and existing problems in traditional Chinese medicine colleges and universities, proposes constructive advices on carrying out information literacy education, moreover, provides a information literacy education standard for traditional Chinese medical colleges and universities and initially constructs a comprehensive information literacy education system.

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