1.Establishing an animal model of delayed onset muscle soreness and its histomorphologic observation
Yuan WEI ; Chunlu FANG ; Liangming LI ; Wenhua XING ; Zeyi YANG
Chinese Journal of Tissue Engineering Research 2015;(29):4645-4651
BACKGROUND:Delayed onset muscle soreness is closely related to skeletal muscle micro-injury, but the exact mechanism of skeletal muscle micro-injury is not yet clear. OBJECTIVE:To observe the histomorphological and ultrastructure changes of skeletal muscle micro-injury models induced by eccentric exercise. METHODS: Forty male Sprague-Dawley rats were randomly divided into quiet control group, immediately after exercise group, post-exercise 24 hours group, post-exercise 48 hours group and post-exercise 72 hours group. In the latter four groups, the rats were subjected to intermittent running on the-16° slope at a speed of 16 m/min: 5 minutes movement, 2 minutes rest and totaly 120 minutes. Rats in the latter four groups were observed immediately, at 24, 48, 72 hours after exercise. RESULTS AND CONCLUSION:After eccentric exercise, the morphology and ultrastructure of rat’s skeletal muscle were both changed to different extents, and Desmin and Vimentin were dyed off for anti-desmin antibody staining at varying degrees. It indicates that one-time eccentric exercise can cause delayed skeletal muscle micro-injury.
2.Effects of corrugated tissue engineered bone scaffold on cell seeding and osteogenesis
Xianli ZENG ; Chunlu YANG ; Jiang LI ; Yuan XU ; Shuo WANG ; Peng WU ; Cuifang WANG ; Yanli DING ; Xinli ZHENG
Chinese Journal of Tissue Engineering Research 2014;(25):3953-3960
BACKGROUND:The effects of engineered bone scaffold containing seeding cels with different shapes to repair bone defect are varied, while the loaded cellquantity is the important factor influencing the curative effect, but which is rarely reported. OBJECTIVE:By preparing self-made corrugated tissue-engineered bone scaffold and other three forms of bone tissue engineering scaffolds, to study the quantity of loaded cels on different scaffolds and osteogenesis of corrugated tissue-engineered bone scaffold so as to discuss the advantages and features of self-made corrugated tissue-engineered bone scaffold. METHODS: (1) Experimentin vitro: There were four kinds of scaffolds with the same volume and samples: calcium phosphate cement (CPC) corrugated surface scaffold group, smooth surface scaffold group, cylindrical scaffold group and porous cylindrical scaffold with holow tubes group, in which the latter three groups are control ones. A certain volume with same density of rabbit bone marrow mesenchymal stem cels (BMSCs) suspension after osteogenesis induction was seeded onto the scaffolds. After incubation, culture, digestion and colection, cellquantity was counted, absorbance value was finaly detected and cellactivity was proofed by alkaline phosphatase and alizarin red staining. (2) Experimentin vivo: New Zealand rabbits were randomly and equaly divided into recombinant human bone morphogenetic protein-2 (rhBMP-2)/CPC/BMSCs corrugated scaffold group, pure CPC corrugated scaffold group and cancelous bone implant group. Three kinds of scaffold implants with the same volume were inserted into the area between rabbit’s L5, 6 transverse processes bilateraly. At 4, 8, 12 weeks postoperatively, gross and histological observation was performed. RESULTS AND CONCLUSION:(1)Experimentin vitro: The drip of cellsuspension steadily stayed on the surface of corrugated scaffold because of corrugated shape groove and the surface tension of the liquid. The amount of cels per sample digested down from the CPC corrugated surface scaffold was significantly more than that from the other three groups (P < 0.05), while the absorbance values did not differ (P > 0.05). (2) Experimentin vivo: At each time point the osteogenesis quantity of rhBMP-2/CPC/BMSCs corrugated scaffold group was more than that of the pure CPC corrugated scaffold group (P < 0.05), while there was no difference from the cancelous bone implant group (P > 0.05). These findings indicate that the characteristics of the self-made corrugated engineered bone scaffold are beneficial to seed cellloading, which supports a large number of osteogenesis and provides feasibility to promote the healing of segmental bone defects.
3.Research progress of osteoporosis based on Th17/Treg balance
Jiayu WANG ; Chunlu YAN ; Fangyu AN ; Lingqing YUAN ; Xiaxia WANG
Chinese Journal of Immunology 2024;40(6):1283-1291
Osteoporosis(OP)is a chronic systemic metabolic bone disease.In recent years,with development of population aging,incidence of OP is also increasing,bringing huge economic burden to family and society.There are many causes of OP,such as aging,oxidative stress,epigenetic and many other aspects,and with further development of research,inflammatory factors and other bone immunology into people's vision.Th17 cells are effector T cell subpopulation that induce development of inflammation and promote bone loss.On the contrary,Treg can maintain their own tolerance and further suppress their own immunity,thus playing a bone protective role.Under physiological conditions,Th17/Treg balance can maintain bone homeostasis,while under pathological condi-tions,disruption of Th17/Treg balance can lead to bone loss.Based on this,this paper will review pathways and cytokines mediating Th17/Treg balance in OP,in order to provide new ideas and methods for treatment of OP.
4.Elucidating the proteomic characteristics and metabolic pathway activation in breast cancer bone metastasis
Yuan CHUNLU ; Chen LONG ; Liao YAFANG ; Shi JIANUO ; Zhang DANDAN
Chinese Journal of Clinical Oncology 2024;51(13):695-702
Objective:To investigate the protein expression characteristics of bone metastatic breast cancer cells and the underlying molecu-lar mechanism driving bone metastasis.Methods:A firefly luciferase-overexpressing human breast cancer cell line,MCF-7-luc,and its bone metastasis subline,MCF-7-BOM-luc,were established.Additionally,a nude mouse model of breast cancer bone metastasis was established by injecting the aforementioned cells into the left ventricle.Bone metastasis and trabecular changes were assessed using micro-computed tomography(Micro-CT).Cell migration,and invasion were evaluated using the Transwell assays.Differential protein expression and epithelial-mesenchymal transition(EMT)were analyzed using proteomics,Western blot,and reverse transcription-quantitative PCR(qPCR).We meas-ured the glucose uptake,L-lactic acid content,and cell energy metabolism parameters using 2-NBDG,ELISA,and a Seahorse energy metabol-izer.Results:MCF-7-BOM-luc cells exhibited stronger migration,invasion,and EMT characteristics as well as more pronounced bone meta-stasis capabilities than the MCF-7-luc cells.Proteomic analysis revealed increased S100 calcium-binding protein A(S100A4)expression in MCF-7-BOM-luc cells,with other differentially expressed proteins being primarily involved in metabolic pathways.Additionally,MCF-7-BOM-luc cells displayed increased expression of E-box binding homeobox 1/2(zinc finger E-box binding homeobox 1/2,ZEB1/2)and Vimentin and reduced E-cadherin expression.MCF-7-BOM-luc cells also exhibited enhanced glucose uptake,L-lactic acid production,and glycolysis rates as well as increased phosphorylation levels of extracellular signal-regulated kinases 1/2(ERK1/2)and signal transducer and activator of tran-scription 3(STAT3).Conclusions:MCF-7-BOM-luc cells promote S100A4 expression through ERK1/2 and STAT3 signaling pathway activation,which in turn enhances the EMT process and glycolysis rate,thereby contributing to malignant bone metastasis.Proteomic analysis of breast cancer bone metastasis-related protein characteristics could offer potential targets for the diagnosis,prevention,and treatment of breast cancer bone metastasis.
5.Research Progress on the Mechanism of Non-coding RNA Regulation of Bone Reconstruction in Osteoporosis and the Therapeutic Mechanism of Traditional Chinese Medicine for Tonifying Kidney and Strengthening Bone
WANG Xiaxia ; AN Fangyu ; YAN Chunlu ; SUN Bai ; WANG Chunmei ; LIU Ying ; SHI Yao ; YUAN Lingqing ; LYU Donghui ; ZHAO Yanzhen
Chinese Journal of Modern Applied Pharmacy 2023;40(17):2462-2472
Non-coding RNAs(ncRNAs) are special RNAs that they don't have protein coding function, but they can affect chromosome structure, gene transcription and participate in the processes of epigenetic modifications. ncRNAs include long non-coding RNAs, microRNA, etc. In recent years, it has been found that these ncRNAs can maintain bone remodeling by adjusting bone resorption and formation in osteoporosis(OP). In the future, it may be a key target of the drug action screening which is clarifying the regulatory mechanism of ncRNAs in the occurrence and development of OP. OP belongs to bone rheumatism category in traditional Chinese medicine, according to the theory of “the kidney generating marrow and dominating bone” in traditional Chinese medicine, kidney tonifying and bone strengthening formulas are used to treat the OP in clinic, and the curative effect is remarkable. It has been found that kidney tonifying and bone strengthening prescriptions can enhance the proliferation of osteoblasts or inhibit the differentiation of osteoclasts by up-regulating or down-regulating the expression of ncRNA, and finally maintain OP bone homeostasis, thus exerting therapeutic effect. However, the specific molecular mechanism is still in its exploratory stage. Therefore, this paper summarized the molecular mechanism of kidney tonifying and bone strengthening prescriptions regulating ncRNAs in the treatment of OP in recent years, in order to provide the new ideas for the screening of the key therapeutic targets of OP drugs and the prevention and treatment of OP with traditional Chinese medicine.