BACKGROUNDLow back pain has emerged as a widespread disease often caused by intervertebral disc degeneration. This study aimed to establish an in vitro cell culture model of rhesus monkey lumbar intervertebral discs and to investigate the effect of combined connective tissue growth factor (CTGF) and tissue inhibitor of metalloprotease-1 (TIMP-1) expression mediated by adeno-associated virus (AAV) on collagen type II and proteoglycan levels. The purpose of these investigations was to explore potential methods for relieving the degeneration of lumbar intervertebral disc cells.

METHODSRhesus monkey lumbar intervertebral disc nucleus pulposus cells (NPCs) were isolated by enzyme digestion, cultured, and transduced with rAAV2-CTGF-IRES-TIMP-1, rAAV2-CTGF, or rAAV2-TIMP-1 at a multiplicity of infection (MOI) of 10(6). The expression of collagen type II and proteoglycan was measured using RT-PCR and Western blotting. The synthetic rate of proteoglycan was measured using (35)S incorporation.

RESULTSRhesus monkey lumbar intervertebral disc NPCs were transduced with rAAV2-CTGF-IRES-TIMP-1, rAAV2-CTGF, and rAAV2-TIMP-1 and the transduced genes were expressed and detected. Compared to the control, CTGF promoted the synthesis of collagen type II and proteoglycan. TIMP-1 showed an enhancing effect on the expression of proteoglycan but no effect on collagen type II. Expression of both genes in rhesus monkey lumbar intervertebral disc NPCs significantly enhances the synthesis of proteoglycan and collagen type II.

CONCLUSIONSSingle gene transduction of CTGF or TIMP-1 can enhanced synthesis of proteoglycan. CTGF expression can also enhance collagen type II protein synthesis. Combined transduction of both CTGF and TIMP1 can significantly promote the expression of proteoglycan and collagen type II to levels greater than transduction of a single gene alone. Our study provides a good basis for multi-gene therapy to treat lumbar intervertebral disc degeneration.

Animals ; Blotting, Western ; Cells, Cultured ; Collagen Type II ; biosynthesis ; Connective Tissue Growth Factor ; genetics ; metabolism ; Intervertebral Disc ; cytology ; Macaca mulatta ; Proteoglycans ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism ; Transduction, Genetic

Background:Early diagnosis and treatment can effectively improve the prognosis of colon cancer.Simple,effective and sensitive screening indicators are of great significance for identification of early cancer and precancerous lesions.L-selectin is a cell adhesion molecule,and podocalyxin(PODXL)is its ligand,both of them play key roles in the development of cancer.Aims:To investigate the expression and significance of L-selectin and its ligand PODXL in colon cancer.Methods:A total of 120 cases of pathological specimens(40 hyperplastic polyp,40 colon adenoma,and 40 colon cancer)from Nov.2020 to Nov.2022 at the Frist People's Hospital of Hangzhou Lin'an District and the First Affiliated Hospital of Zhejiang Chinese Medical University were collected,and 20 cases of normal intestinal mucosal tissue were served as controls.qRT-PCR and immunohistochemistry were used to detect the mRNA and protein expressions of L-selectin and PODXL,respectively.Western blotting was used to determine the expressions of L-selectin and PODXL,and their relations with different clinicopathological parameters of colon cancer were analyzed.In addition,60 serum specimens of colon cancer were collected.ELISA was used to detect serum concentrations of L-selectin and PODXL.Results:Expressions of L-selectin and PODXL mRNA and protein in colon adenoma group were significantly higher than those in normal controls and hyperplastic polyp group(P<0.05),and mRNA and protein expressions of L-selectin and PODXL in colon cancer group were significantly higher than those in normal controls,hyperplastic polyp group and colon adenoma group(P<0.05).Significant differences in protein expressions of L-selectin and PODXL were found in different pathological types,lymph node metastasis,Dukes staging in colon cancer(P<0.05).Expression of L-selectin was positive correlated with expression of PODXL in colon cancer(r=0.855,P<0.001).Serum concentrations of L-selectin and PODXL were significantly lower in the initial group than in the relapse group(P<0.05),and serum concentrations of L-selection and PODXL was significantly lower in the non-metastatic group than in the metastatic group(P<0.05).Serum concentrations of L-selectin and PODXL at 3 months after surgery was significantly lower than 3 days after surgery and before surgery(P<0.05).Conclusions:L-selectin and PODXL may be involved in the development and progression of colon cancer.They are carcinogenic proteins,and their detection could provide reference value for the prevention and early diagnosis of colon cancer,and through early screening of lesion could improve the prognosis of colon cancer to a certain extent.

Background:Studies have confirmed that colorectal tumors may be caused by normal mucosa near or inside the diverticulum,and patients with colon diverticulum are more likely to develop colon cancer than those without diverticulum.When colorectal adenoma involves diverticulum,the inflammatory state in the diverticulum will increase the risk of adenomatous dysplasia.Therefore,adenomas involving colon diverticulum are more likely to develop malignant changes,and adenomas involving colon diverticulum need to be resected to avoid colon cancer outcomes.Improved prognosis.Aims:To explore the efficacy and safety of endoscopic mucosal resection(EMR)treatment of colorectal tumors near or involving a diverticulum.Methods:Fourteen consecutive cases of adenoma(near or involving a diverticulum)during EMR treatment from Jun.2018 to Jan.2022 were collected,and analyze their clinical characteristics and outcomes.The main outcomes were adverse events,including bleeding,perforation,and electrocoagulation syndrome,while the secondary outcome was the total resection rate,block resection rate and local recurrence rate.Results:Among the 14 patients with colon adenoma,1 case was inside the diverticulum,13 cases were near the diverticulum,the diameter of the lesion was(0.76±0.25)cm,the operation time was(19.6±5.33)min.The main tissue types were tubular adenomas,and the pathologic characteristics were low grade intraepithelial neoplasia.Among the complications,delayed hemorrhage was found in 1 case(7.1%),the patient had a history of taking antiplatelet drugs(clopidogrel),electrocoagulation syndrome was found in 1 case(7.1%),and appendiceal orifice and diverticulum were involved in this patient,with no perforation event.The whole resection rate and the complete resection rate were 100%(14/14).Within 1 year after surgery,10 patients came to the hospital for re-examination of colonoscopy,and no local recurrence was found.Conclusions:EMR treatment involving colonic diverticulum adenoma is safe and effective.However,patients with adenoma involving antiplatelet drugs and appendiceal orifice and diverticulum should be alert to postoperative complications.