1.Compound 3k for osteoarthritis:mechanism of modulating oxidative stress pathway to improve chondrocyte glycolysis
Surong GUO ; Shisheng CAO ; Xingtong MU ; Qing YANG ; Juan ZHANG
Chinese Journal of Tissue Engineering Research 2025;29(2):363-370
BACKGROUND:Osteoarthritis is now considered a metabolic disease.Previous studies have shown that glycolysis plays an important role in the occurrence and development of osteoarthritis.Compound 3k,as a novel small molecule inhibitor of glycolysis,has anti-inflammatory and anti-tumor effects.Therefore,it can target glycolysis and is expected to provide new ideas for the treatment of osteoarthritis. OBJECTIVE:To explore the role of Compound 3k in osteoarthritis caused by glycolytic overactivity based on the hypoxia-inducible factor 1 alpha(HIF-1α)/reactive oxygen species(ROS)pathway. METHODS:ATDC5 chondroblasts at logarithmic growth phase were taken to induce osteoarthritis in an in vitro cellular model by the action of 10 ng/mL interleukin-1β for 24 hours.The cytotoxicity of Compound 3k at different concentrations(0.25,0.5,1,2.5,5,10,15 μmol/L)was detected by cell counting kit-8 assay,and the appropriate concentrations were selected for the subsequent experiments.The chondrocytes were randomly divided into control,model and treatment groups.The model group was induced with 10 ng/mL interleukin 1β,and the treatment group was pre-stimulated with Compound 3k for 2 hours and then co-cultured with interleukin 1β.The proliferation of the cells in each group was detected by the cell counting kit-8 assay;the inflammatory level of the cells in each group was detected by the ELISA kit;the ROS,extracellular lactate and glucose contents were detected using the kit;qRT-PCR and western blot were used to detect the levels of related inflammatory factors,interleukin-6 and tumor necrosis factor-α,glycolysis-related genes glucose transporter protein-1,glyceraldehyde 3-phosphate dehydrogenase,monocarboxylate transporter protein-1 and HIF-1α. RESULTS AND CONCLUSION:Compared with the control group,the model group showed a decrease in cell proliferative activity,active glycolysis level,manifested by an increase in extracellular lactate content(P<0.001)and a decrease in glucose content(P<0.001),interleukin-6(P<0.000 1)and tumor necrosis factor-α(P<0.001).The expression levels of glycolysis-related genes glucose transporter protein-1(P<0.001),glyceraldehyde 3-phosphate dehydrogenase(P<0.001),monocarboxylic acid transporter protein-1(P<0.001)and HIF-1α(P<0.001)in the model group were all up-regulated,accompanied by oxidative stress and overproduction of ROS.Compared with the model group,Compound 3k treatment effectively increased cell proliferation activity and inhibited the level of overactive glycolysis(P<0.001),while suppressing the expression of genes related to inflammation(P<0.001)and glycolysis in osteoarthritic chondrocytes,inhibiting oxidative stress,downregulating the expression level of HIF-1α(P<0.000 1)and decreasing the content of ROS.To conclude,Compound 3k inhibits interleukin-1β induced chondrocyte inflammation,and its mechanism may be related to glycolysis and HIF-1α/ROS mediated oxidative stress.
2.iTRAQ-based proteomic analysis of hippocampus of pentylenetetrazol-kindled epileptic rats
Peng ZHANG ; Li ZHANG ; Hongli XIONG ; Shisheng ZHU
Chinese Journal of Neuroanatomy 2023;39(6):649-656
Objective:To screen differentially expressed proteins(DEPs)in the hippocampus of epileptic rats,pro-viding ideas for further exploring the pathogenesis and drug therapeutic targets of epilepsy.Methods:The epileptic rat model(PTZ group)induced by pentylenetetrazol(PTZ).The protein profiles of hippocampal tissues of the PTZ group and control groups were detected by isobaric tag for relative and absolute quantitation(iTRAQ)combined with LC-MS/MS technology,and the fold changes in the protein expression of the PTZ group versus the control group was considered to be more than 1.5 or less than 0.67,and P<0.05 was taken as the criteria for screening DEPs.Then DEPs were subjected to bioinformatics analyses such as Gene Ontology(GO)functional annotation and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.Results:A total of 80 proteins showed significantly different expression,which included 39 up-regulated and 41 down-regulated.GO analysis showed that the up-regulated DEPs were mainly involved in biological processes such as cellular response to nerve growth factor stimulus,axon develop-ment,cell surface receptor signaling pathway,neuron migration,actin filament depolymerization,and signal transduc-tion,while down-regulated DEPs were mainly involved in biological processes such as tricarboxylic acid cycle,citrate metabolism process,acetyl-CoA biosynthetic process from pyruvate,oxaloacetate metabolism process,and regulation of focal adhesion assembly.KEGG pathway enrichment analysis showed that up-regulated DEPs were mainly involved in five signaling pathways,including AMPK signaling pathway,regulation of actin cytoskeleton,sphingolipid signaling pathway,phenylalanine metabolism,and insulin signaling pathway;The down-regulated DEPs were mainly involved in six signaling pathways,including the citric acid cycle,carbon metabolism,acetate and dicarboxylate metabolism,meta-bolic pathways,amino acid biosynthesis and amyotrophic lateral sclerosis signaling pathways.Conclusion:DEPs from epileptic hippocampal tissues were screened by iTRAQ proteomics,and the metabolic pathways enriched by bioinformat-ics methods for DEPs may be closely related to the pathogenesis of epilepsy.
3.Distinguishing subgingival plaques of patients with chronic periodontitis and type 2 diabetes mellitus based on Raman spectroscopy
Juan ZHANG ; Yiping LIU ; Shisheng CAO ; Xin LI ; Xiaoxi DONG ; Hongxiao LI
Journal of China Medical University 2023;52(12):1113-1118
Objective The aim of this study is to combine Raman spectroscopy and machine learning techniques to distinguish subgin-gival plaques among three groups of subjects,including patients with chronic periodontitis(CP)and type 2 diabetes mellitus(T2DM),patients with CP alone,and healthy controls.Methods The Raman spectra of the subgingival plaques from 20 patients with CP and T2DM(group A),23 patients with CP alone(group B),and 23 healthy controls(group C)were obtained using a portable Raman spec-trometer.Eight common machine learning algorithms were applied to build models to distinguish the Raman spectra of the three types of subgingival plaques.Results The model identified as optimal for distinguishing the three types of subgingival plaques was linear discri-minant analysis(LDA).The optimal model to distinguish groups A and B is LDA,groups A and C is extra trees(ET),and groups B and C group is LDA.Conclusion The proposed classification model based on Raman spectroscopy and machine learning algorithms can dis-tinguish subgingival plaques among patients with CP and T2DM,with CP alone,and healthy controls.This technique can be used in future clinical practice as a screening or diagnostic tool.
4.Rapid screening of single guide RNA targeting pig genome and the harvesting of monoclonal cells by microarray seal.
Mengyu GAO ; Xinglong ZHU ; Shisheng WANG ; Bingqi ZHANG ; Yunlin ZHANG ; Yuting HE ; Yanyan ZHOU ; Shun LI ; Guang YANG ; Guangneng LIAO ; Ji BAO ; Hong BU
Journal of Biomedical Engineering 2021;38(1):111-121
The emergence of regular short repetitive palindromic sequence clusters (CRISPR) and CRISPR- associated proteins 9 (Cas9) gene editing technology has greatly promoted the wide application of genetically modified pigs. Efficient single guide RNA (sgRNA) is the key to the success of gene editing using CRISPR/Cas9 technology. For large animals with a long reproductive cycle, such as pigs, it is necessary to screen out efficient sgRNA
Animals
;
CRISPR-Cas Systems/genetics*
;
Clustered Regularly Interspaced Short Palindromic Repeats/genetics*
;
Gene Editing
;
RNA, Guide/genetics*
;
Swine
5.Effect of long needle penetration needling on matrix metalloproteinase inhibitor 1 in the synovium of knee osteoarthritis model rats
Shisheng LI ; Yaochi WU ; Junfeng ZHANG
Chinese Journal of Tissue Engineering Research 2016;20(18):2609-2614
BACKGROUND: Matrix metal oproteinase inhibitor 1 plays an important role in the inhibition of articular cartilage degeneration and destruction.
OBJECTIVE: To observe the changes of matrix metal oproteinase inhibitor 1 expression level in synovium of knee osteoarthritis rats after treated with penetration needling of long needle.
METHODS: A total of 40 Sprague-Dawley rats were randomly assigned into normal group, penetration needling group, drug group and model group, with 10 rats in every group. 1.6% papain solution was injected into knee joint cavity of Sprague-Dawley rats. 2 weeks later, knee osteoarthritis models were established. Rats in the normal and model groups did not receive any intervention. In the penetration needling group, rats were treated by penetration needling with 0.3 mm×125.0 mm filiform needle. Rats in the drug group were intraarticularly injected with sodium hyaluronate.
RESULTS AND CONCLUSION: (1) Enzyme linked immunosorbent assay: at 4 weeks after treatment, matrix metal oproteinase inhibitor 1 expression in synovial membrane was higher in the normal group than in the model group (P < 0.05). Matrix metal oproteinase inhibitor 1 expression in synovial membrane was higher in the penetration needling group and drug group than in the model group (P < 0.05). (2) Results verified that long needle penetration therapy can effectively increase matrix metal oproteinase inhibitor 1 expression in synovial membrane of knee osteoarthritis model rats.
6.Predictive value of combined detection of serum tumor markers for recurrence or metastasis of colon cancer after radical surgery
Yu ZHANG ; Xin LI ; Wen LUO ; Tingting NI ; Shisheng TAN
Chongqing Medicine 2015;(26):3616-3617,3621
Objective To explore the clinical application value of the expression levels of serum hypoxia induced factor-1α(HIF-1α),vascular endothelial growth factor(VEGF),carcino-embryonic antigen(CEA)and CA125 in predicting the recurrence or metastasis of colon cancer after radical resection.Methods Totally 123 cases of colon cancer after radical surgery in our hospital from July 2009 to December 2013 were performed the retrospective analysis.The correlation between the serum markers levels with the recurrence or metastasis was dynamically analyzed.Results (1)The HIF-1αand CEA levels after operation in 123 cases of co-lon cancer were decreased significantly compared with before operation;(2)The HIF-1α,CEA and CA125 levels in 39 cases of re-currence or metastasis were significantly increased compared with those in 84 cases of non-recurrence or non-metastasis (P <0.05);(3)in 39 cases of recurrence or metastasis,the HIF-1α and CEA levels in recurrence or metastasis were increased signifi-cantly compared with those after the end of chemotherapy (P <0.05);(4)HIF-1αwas positively correlated to VEGF(P <0.05). Conclusion The serum levels of HIF-1αand CEA in the patients with colon cancer are positively correlated with recurrence or me-tastasis,the dynamically combined detection of serum markers has an important significance for predicting and early find recurrence or metastasis after radical operation in colon cancer.
7.An improved method for percutaneous pedicle screw implantation and the effect of the improvement on radiation exposure
Xu ZHOU ; Shisheng HE ; Hailong ZHANG ; Guangfei GU ; Lei ZHANG ; Qingsong FU
Chinese Journal of Orthopaedics 2014;34(3):265-272
Objective To introduce a new minimally invasive lumbar positioning system and its positioning method as well as conduct a prospective control study on the differences in positioning time,positioning frequency and radiation exposure dose between the new positioning method and the traditional positioning method.Methods 121 patients with lumbar disc herniation combined with lumbar instability and patients with thoracolumbar vertebral fracture admitted in our hospital from May 2010 to February 2013 were randomized into two groups.68 patients in Group A had undergone the traditional positioning method before and during operation.Among the 68 patients,41 were with single-segment lesion and 27 were with two-segment lesion.53 patients in Group B had undergone the new minimally invasive lumbar positioning system before and during operation.Among the 53 patients,34 were with single-segment lesion and 19 were with two-segment lesion.There were no significant differences in gender,age,course of disease and other general information between the two groups.The positioning time,positioning frequency and radiation dose of the two groups before and during operation were compared and statistically analyzed.Results The positioning time of Group A and Group B were 8.26±3.44 min and 3.51±1.82 min respectively; the positioning frequencies were 3.57 and 1.22; the accuracy were 60.8% and 96.2%.For the patients with single-segment lesion,the positioning time before implanting two percutaneous pedicle screws during operation were 15.12±4.69 min and 5.51±1.32 min respectively and the positioning frequency were 6.47 and 2.45.For the patients with two-segment lesion,the positioning time before implanting three percutaneous pedicle screws during operation were 24.91±7.43 min and 8.84±2.32 min respectively and the positioning frequency were 11.72 and 3.69.Moreover,the radiation dose detected at neck,chest and wrist of the surgical staff in Group A were 3.09±0.24 Gy,4.23±0.71 Gy and 5.17±0.62 Gy and that detected in Group B were 1.38±0.47 Gy,2.69±0.33 Gy and 3.21±1.05 Gy.There were significant differences in positioning time,positioning frequency and radiation dose between Group A and Group B.Conclusion The minimally invasive spine positioning system and positioning method can simplify the operative procedures and largely reduce radiation exposure,which is characterized by high positioning accuracy,short positioning time,low fluoroscopy frequency and X-ray radiation dose.
8.Therapeutic Observation on Tuina plus Electroacupuncture for Lateral Humeral Epicondylitis
Shisheng LI ; Yaochi WU ; Junfeng ZHANG ; Jinghui ZHOU
Journal of Acupuncture and Tuina Science 2014;(5):321-325
Objective: To observe the clinical efficacy of tuina combined with electroacupuncture (EA) in treating lateral humeral epicondylitis. <br> Methods: Totally 118 patients were randomly allocated into a treatment group and a control group, 59 cases in each. The patients in the treatment group were treated with tuina combined with EA treatment, while those in the control group were treated by EA alone. Clinical efficacies of the two groups were compared after the treatment. <br> Results: The total effective rate was 93.2% in the treatment group, versus 84.7% in the control group, and the difference was statistically significant (P<0.05). <br> Conclusion: Tuina combined with EA treatment for lateral humeral epicondylitis has a significant effect.
9.Common complication and progress of Percutaneous endoscopic lumbar discectomy
Shan ZHAO ; Hailong ZHANG ; Xin GU ; Xiaofei GUAN ; Shisheng HE
International Journal of Surgery 2014;41(11):780-783
Percutaneous endoscopic lumbar discectomy belongs to minimally invasive spine operation Its superiority includes smalleroperation wound,less bleeding,shorter hospital day,and earlier return to function,conpared with the traditional operation.At the same time,percutaneous endoscopic lumbar discectomy has complications,as the open operation.This paper reviews its common complications,diagnosis,prevention and control.
10.Minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) with unilateral pedicle screw fixation in treatment of one-level lumbar degenerative disease
Xiaolong SHEN ; Hailong ZHANG ; Xin GU ; Shisheng HE
Chinese Journal of Orthopaedics 2014;34(7):749-755
Objective To explore the clinical and radiographic outcomes of minimally invasive transforarninal lumbar interbody fusion (MIS-TLIF) with unilateral pedicle screw fixation in treatment of one-level lumbar degenerative disease.Methods A total of 65 patients suffered from one-level lumbar degenerative disease between October 2009 and December 2011.They were divided into 2 groups according to different fixation ways.31 patients were given MIS-TLIF with unilateral pedicle screw fixation.The other 34 patients were given MIS-TLIF with bilateral pedicle screw fixation.Microscopic tubular retractor system (METRxMD) and Sextant system were used in all the procedures of treatment.The whole lumbar lordosis (WL),the segmnental lordosis (SL),fusion level disc space angle,lumbar scoliosis angle,and segmental scoliosis angle were measured pre and post operation according to standarded X-rays.The disc height index (DI) and the lumber curvature index (LI) were also evaluated.The Oswestry disability index (ODI) score and visual analog scale (VAS) pain score data were obtained from all the patients pre-operation and during each following-up procedure.Results All the patients were well followed up 18 months to 36 months(average 26.6 months).All the 65 patients were proved to achieve bone fusion in 12 months post-operation.The ODI and VAS scores post-operation improved significantly in each group,but showed no significant difference between the 2 groups.Likewise,there were no significant differences between the 2 groups in datas of WL,SL,fusion level disc space angle,lumbar scoliosis angle,segmental scoliosis angle,DI,and LI.But there was a positive linear correlation between the LI and WL in the 2 groups.Conclusion MIS-TLIF with unilateral pedicle screw fixation is as good as MIS-TLIF with bilateral pedicle screw fixation in patients of one-level lumbar degenerative disease.

Result Analysis
Print
Save
E-mail