1.Brain-derived neurotrophical factor after olfactory ensheathing cells transplantation in spinal cord injury of rats
Lifa CHEN ; Jieyuan ZHANG ; Zhaoxia DUAN ; Bingcang LI ; Huarong YU
Chinese Journal of Organ Transplantation 2011;32(5):296-299
Objective To observe the expression of brain-derived neurotrophical factor (BDNF) in injury spinal cord after transplantation olfactory ensheathing cells (OECs), and to investigate the mechanism of OECs repairing spinal cord injury.Methods OECs from GFP transgenic rats were separated and cultured for transplantation. Spinal cord injury rats were separated two groups by random digits table. In experimental group, OECs suspension were transplanted into injured spinal cord following spinal cord injury. In control group, DMEM was transplanted into the injured spinal cord after spinal cord injury. Motor function was evaluated per week after transplantation. The expression levels of BDNF mRNA and protein were detected by using RT-PCR and immunohistochemistry respectively, and compared with those from normal SD rats.Results Motor function of two groups was improved gradually after transplantation. The motor function scores in experimental group was obviously higher than in control group at 21st day after transplantation (P<0.05). A lot of survival GFP OECs distributed around impaired myeloid tissue. At 21st day after transplantation, BDNF mRNA and protein expression in experimental group were strongest (P<0.05), and stronger in control group than in normal group (P<0.05).Conclusion The transplantation of OECs can repair the injured spinal cord by increasing the expression of BDNF mRNA and protein to improve local microenvironment.
2.Distrabution and migration of olfuctory ensheathing cells transplanted into the contused spinal cord of rats
Yue LI ; Hualin YU ; Lifa CHEN ; Jieyuan ZHANG ; Bingcang LI
Chinese Journal of Trauma 2011;27(1):78-82
Objective To observe the migration and distribution of OECs in injured spinal cord and discuss their relation with the recovery of spinal cord function. Methods The rats were contused by a force of 10 g · 25 mm with NYU-impactor at T10 level. The OECs acutely isolated from green fluorescence protein (GFP) rats were purified, identified and then transplanted into the injured site and the rostral and caudal parts of the spinal cord one week after injury, with total volume of the transplanted OECs for 90 000/μl. Within 13 weeks after transplantation, the migration and distribution of OECs were qualitatively observed on the cryo-sections under fluorescence light microscope. The area and the length of OECs distribution were semi-quantitatively determined. The locomotor function of the spinal cord was appraised by BBB score. Results OECs were located collectively in the transplanted site at early stage after transplantation and then spread gradually mainly along the long axis of the cord. OECs could be found in the cavity of the contused spinal cord. The area and the length of OECs distribution were increased from 1.33 mm2 and 4.23 mm respectively at one week to 3.30 mm2 and 7.68 mm respectively at 13 weeks after transplantation. In the meantime, the locomotor function was gradually improved. Conclusion OECs can migrate within the contused spinal cord, as may contribute to the recovery of locomotor function.
3.The expression of RCN3 in colon cancer and its clinical significance
Songlin HOU ; Qiang PENG ; Yu ZHOU ; Jia CHEN ; Xingjiang XIE ; Lifa LI ; Tong ZHOU ; He ZHOU
Journal of Chinese Physician 2022;24(5):712-718
Objective:To analyze the expression and clinical significance of reticulocalbin 3 (RCN3) in colon cancer by bioinformatics database and biological experiments.Methods:Colon cancer HT29 and SW620 cells and colon normal mucosal cells FHC were cultured. The expression of RCN3 in cells was verified by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blot. The expression data of RCN3 in normal colon tissue and colon cancer tissue were obtained by Ualcan database. The co-expressed gene information of RCN3 from LinkedOmics database was obtained, and the biological processes and related functions of these RCN3 co-expressed genes through were analyzed by gene ontology analysis (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The protein-protein interaction network of RCN3 related coding genes was constructed by using STRING database. Finally, the relationship between the expression of RCN3 and the clinical prognosis of patients with colon cancer was compared and analyzed according to GEPIA, Ualcan and Linked Omics biological database.Results:Western blot and qRT-PCR results showed that the mRNA and protein expression of RCN3 in HT29 and SW620 colon cancer cells was significantly higher than those in FHCcells ( all P<0.05). The analysis of biological database showed that the expression level of RCN3 in colon cancer tissue was higher than that in normal colon tissue ( P<0.05). GO enrichment analysis showed that RCN3 co-expression genes were mainly involved in the composition of extracellular matrix and extracellular domain structure, the binding process of extracellular matrix and multiple receptors, and the biological processes related to tumor development such as cell adhesion, immune response, and angiogenesis through extracellular domain structure. KEGG pathway analysis showed that RCN3 co-expression genes were mainly involved in ECM receptor interaction, cytokine receptor interaction, chemokine signaling pathway, phosphatidylinositol-3-kinase protein kinase B (PI3K-Akt) signaling pathway, phagosome signal, IgA related intestinal immune network signal, these signaling pathways always related to tumor invasion, migration and inflammatory immune response. The protein-protein interaction network analysis showed that the coding protein genes that directly interacted with RCN3 protein that included PRDX6, NOSIP, PCSK6, IMMP1L, PRRG2, FBXO47, FCGRT, FKBP9, PCDHGA12, and PNMAL1, which were mainly involved in the occurrence and development of colorectal cancer, liver cancer, gastric cancer, breast cancer, lung cancer, and ovarian cancer. Survival curve analysis showed that the overall survival rate of colon cancer patients with high expression of RCN3 was significantly lower than that of patients with low expression of RCN3 ( P<0.05). Conclusions:RCN3 is highly expressed in colon cancer tissues and cells, which is closely related to the occurrence, development and prognosis of colon cancer. It can be used as one of the markers for early screening and prognosis prediction of colon cancer.