Background/Aims: Cholangiocarcinoma (CCA) is a primary malignant neoplasm with an extremely poor prognosis. While combined chemoradiotherapy has been demonstrated to delay CCA progression to a certain extent, the absence of specific molecular biomarkers or targets significantly hinders the diagnosis and treatment of CCA. Methods: Through cross-analysis of proteomics and ADMA modificationomics, we identified DDX1 overexpressed in CCA with elevated R602-ADMA modifications. HPLC-MS/MS identified PRMT1 as the methyltransferase and USP10 as the deubiquitinating enzyme for DDX1. Immunofluorescence and nuclear-cytoplasmic partitioning experiments confirmed DDX1’s nuclear localization. GO and KEGG analyses clarify the biological functions of DDX1 in response to hypoxia. RNA-seq transcriptomics analyzed key pathways influenced by DDX1. A hydrodynamic in situ CCA mouse model was established to validate the chemopreventive effects of the PRMT1-specific inhibitor GSK715 on CCA development. Results: DDX1 promotes CCA progression both in vivo and in vitro and can be inhibited by GSK715. Mechanistically, PRMT1 mediates ADMA modification at position R602 of DDX1. This modification promotes DDX1 nuclear localization by recruiting USP10 to deubiquitinate DDX1, while simultaneously inhibiting PRMT1 degradation. DDX1 promotes the transcription of PRMT1 and USP10 by binding to the mRNA 3’UTR region, establishing a positive feedback regulatory pathway. This mechanism promotes the occurrence and development of CCA and can serve as a target for the inhibitor GSK715 to suppress CCA progression. Conclusions Our study identified DDX1-R602-ADMA modification as a novel ADMA modification in CCA. It further confirmed its pivotal role in CCA progression. Targeting the USP10-PRMT1-DDX1 axis may represent a significant therapeutic approach for CCA.

Objective To make sciatic nerve traction injury models of rabbit , in order to prospectively evaluate possibility and accuracy of diffusion tensor tracking (DTT) in sciatic nerve injures. Methods The right sciatic nerves of 32 New Zealand white rabbits were selected for traction injury , and the left sciatic nerves were the sham operation side. DTI scan was performed before and after the operation on 1st day , 3rd day, and 1 week, 2, 3, 4, 6 and 8th week respectively, and DTT was reconstructed. Then the length of reconstructed fiber tracts and fiber density index were calculated. After the MRI scan , the sciatic nerve was removed to perform pathologic examination at different time points. Results The difference of the length of reconstructed fiber tracts between nerve traction injury and sham operation side was significant at 1 day~2weeks after operation (P<0.05), while the difference was not significant at 3~8 weeks.The fiber density index of nerve traction injury and sham operation side was significantly different at 1 day~8 weeks(P<0.05). 1day after operation, myelin sheath of traction portion was obviously twisting. 3 days after operation , a large amount of myelin sheath broke down. 2 weeks after operation , axon, myelin sheath degeneration and regeneration coexisted at the same time. 8 weeks after operation , nerve fibers regenerated and restored to normal structure. Conclusion The length time curve and density index-time curve of nerve traction injury are consistent with the changes of pathology , which can be used as a supplementary method to evaluate the degeneration and regeneration in nerve injury.

Objective To investigate the correlation between eigenvalues(λ∥ andλ⊥)of sciatic nerve and limb function in a rabbit model of acute nerve traction inj ury for finding the role ofλ∥ andλ⊥ on diagnosis of nerve inj uries.Methods The right sciatic nerves of 32 Newland rabbits were selected to be performed as traction injury,and then left sciatic nerves were treated as control for the sham operation side.MRI DTI scan was performed before and after the operation on 1 day ,3 day,and 1 week,2,3,4,6 and 8 weeks respectively.Theλ∥ andλ⊥ of inj ured sciatic nerves and sham operated nerves were measured,meanwhile functional exami-nations were evaluated,and then analyzing the correlation betweenλ⊥ of inj ured sciatic nerves and the score of inj ured limb function in the periods of 3 days to 8 weeks;finally different portions of sciatic nerve were removed for histological examinations.Results Theλ⊥ of inj ured sciatic nerves in the proximal portion was slightly increased at 3 days and recovered to the normal at 2 weeks.Theλ⊥ of inj ured sciatic nerves in the traction portion and distal portion were slightly raised at 1 day,reached to its maximum value at 3 days,and then decreased gradually after 1 week and dropped to the pre-operation level after 4 weeks.Theλ⊥ of inj ured sciatic nerves in the proximal portions was significantly higher than that of the sham-operated nerves from 1 day to 1 week(F=7.275,P=0.000). Theλ⊥ of inj ured sciatic nerves in the traction portion and distal portion were significantly higher than that of the sham-operated nerves from 1 day to 3 weeks(F=5.851,F=3.794,P=0.001,P=0.000).However,there was no significantly difference between theλ∥ of inj ured nerves in each portion and sham-operated nerves .The negative correlation between theλ⊥ of inj ured sciatic nerves in the distal portion and the score of the nerve function in the injured limb was found (r=-0.938,r=-0.897,P<0.01).Conclu-sion Theλ⊥ of inj ured nerves in the traction portion and distal portion could monitor the process of degeneration and regeneration of sciatic nerve in traction inj ury,while theλ∥ plays no obvious role in diagnosis of nerve inj uries.

This paper is aimed to investigate the feasibility of applying the small intestine submucosa (SIS) as the scaffold in constructing tissue engineering cartilage in vitro. We obtained SIS from the small intestine of specific pathogen-free pigs. Then we isolated tunica submucosa layer from the mucosal, muscular, and serosal layers by gentle mechanic abrasion. The SIS was made acellular by combination of detergent and enzyme digestion. The chondrocytes were seeded onto the SIS and were cultured for 3 weeks. The cell growth, attachment and distribution were detected by histochemical stain, immunohistochemical stain and scan electron microscope. The chondrocytes could adhere and grow well on the matrix surface, and synthesize a large of the GAG and type U collagen. However, the chondrocytes grew only on the surface andsuperficial layer of the scaffold, they did not move into the inner part of the scaffold. It could be concluded that SIS has good cellular compatibility without cytotoxicity and provides temporary substrate to which these anchorage-dependent cells can adhere, and stimulate the chondrocytes anchored on the scaffold to proliferate and keep differentiated phenotype. Further study will be needed to promote the ability of chondrocyte chemotaxis in order to distribute the chondrocytes into the whole scaffold uniformly.
Animals ; Cell Adhesion ; Cell Culture Techniques ; Cell Proliferation ; Chondrocytes ; cytology ; Chondrogenesis ; physiology ; Intestinal Mucosa ; cytology ; Intestine, Small ; cytology ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds