ABSTRACT
As the main stabilizing structure of the medial ankle joint, deltoid ligament plays a role in counteracting excessive eversion of the hindfoot and external rotation of the talus during ankle movement so as to maintain the biomechanical stability of ankle joint. Although the incidence of deltoid ligament injury is low, improper diagnosis and treatment can affect the path of talus motion and eventually lead to chronic medial instability or traumatic arthritis of the ankle joint, seriously affecting the normal life and motor function of the patients. The diagnosis of deltoid ligament injury needs to be based on the characteristics of the injury, physical examination and imaging, among which X-ray, MRI and ultrasonography are most frequently used. There are various methods to treat deltoid ligament injury according to the type of injury, and thus the choice of treatment has been a hot topic in the field of foot and ankle surgery. The choice of non-surgical or surgical treatment for acute deltoid ligament injury remains controversial. For the treatment of chronic deltoid ligament injury, there is no consensus on direct repair or deltoid ligament reconstruction. In addition, the choice of autologous or allograft tendon or wire anchors for deltoid ligament reconstruction is also disputed. The rehabilitation of deltoid ligament injury is crucial to the early restoration of motor function of the ankle joint, but the related guidelines or consensus are scarce. In order to fully understand the characteristics of deltoid ligament injury, make accurate diagnosis and formulate reasonable treatment and rehabilitation programs, the authors review the research progress in deltoid ligament injury from aspects of anatomical characteristics, biomechanical mechanism of injury, diagnosis, treatment and postoperative functional rehabilitation, hoping to provide a reference for the clinical diagnosis and treatment of deltoid ligament injury.
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Objective@#To investigate the effect and mechanism of LncRNA ANRIL on the radiosensitivity of HCT116 cells line and nude mouse transplant tumors.@*Methods@#The expression of LncRNA ANRIL in colorectal cancer cells was detected by qPCR. The negative control siRNA, ANRIL siRNA, miR-NC mimic, miR-195 mimic, miR-NC inhibitor and miR-195 inhibitor were transfected into HCT116 cells, and marked as negative control group, silencing ANRIL group, overexpressing miR-NC group, overexpressing miR-195 group, inhibiting miR-NC group and inhibiting miR-195 group, and the HCT116 cells without any treatment were marked as the blank control group. The clone formation assay was used to detect radiosensitivity of colorectal cancer cells, flow cytometry was used to detect apoptosis. The web site, StarBase, was used to predict the downstream miRNAs of ANRIL and dual luciferase reporter gene assay was used to further verify. Subcutaneous tumor transplantation assay was used to detect the effect of ANRIL on the growth of colorectal cancer cells after irradiation.@*Results@#After irradiation with 2, 4, 6 and 8 Gy, the cell survival fraction of silencing ANRIL group was significantly decreased when compared with that of negative control group (P<0.05), and the radiosensitivity ratio was 1.52. The apoptosis rate of the silencing ANRIL+ 4 Gy group was significantly higher than that of the negative control+ 4 Gy group ((27.86±2.78)% vs. (12.06±1.46)%, P<0.05). The results of the experiment on nude mouse transplant tumors showed that the tumor volume in the negative control group was lower than that of the silent ANRIL group on days 13, 16, 19, 22 and 25 ((234±66) mm3, (273±63) mm3, (296±72) mm3, (321±85) mm3 and (403±94) mm3 vs. (357±79) mm3, (485±124) mm3, (617±143) mm3, (764±174) mm3 and (985±221) mm3P<0.05). MiR-195 is a target gene of ANRIL, and inhibition of miR-195 can reverse the inhibitory effect of silencing ANRIL on radiosensitivity, apoptosis and xenografts of HCT116 cells.@*Conclusions@#LncRNA ANRIL regulates the radiosensitivity of colorectal cancer cells by miR-195, which may provide a new sensitizing target for clinical colorectal cancer radiotherapy.
ABSTRACT
Aim To develop an in vitro high throughput drug screening system based on reporter gene assay for identification of novel compounds with PXR, FXR and LXRα agonist activity. Methods The expressions of exogenous PXR, FXR and LXRαgene in HEK293, HepG2 and LS174T cells were examined by Real-Time quantity PCR. pSG5-hPXR and pGL3-XREM-CYP3A4, pEGFP-N3-hFXR and EcRE-TK-Luc, pCMX-FLAG-hLXRα and pGL3-XREM-CYP3A4 were cotransfected into cells and the optimal ratio of three plasmids was determined. The dose-response relationship between the positive drug and the fold induction was determined. The specificity of the model was ex-amined, and the repeatability was also determined by Z′ value. Results ① The PXR, FXR and LXRα mRNA expression in HEK293 cell is low among three different cells. ②reporter gene vector and expression plasmid ratio of 1∶ 1, 2∶ 1 and 2∶ 1 were proved to be suitable for highest relative luciferase activity for PXR, FXR or LXRα agonist screening model. ③ The relative luciferase activity was induced by Rif, CDCA or T0901317 in a dose-dependent manner. ④Only Rif, CDCA or T0901317 could significantly increase the relative luciferase activity in PXR,FXR or LXRα agonist screening model, no effect of other nuclear re-ceptors agonist was observed, and the values of Z′-factor for PXR, FXR and LXRαagonist screening model were 0. 58, 0. 66 and 0. 63, respectively. Conclusion An in vitro PXR, FXR and LXRα agonist high-throughput screening models are devel-oped with acceptable specificity and repeatability, and the mod-els can be used to screen PXR, FXR and LXRα agonist.
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Objective:To investigate the effect of scutellarin on P-gp protein expression and activity in Caco-2 cells. Methods:Scutellarin(25,50 and 100 μmol·L-1 )was incubated with Caco-2 cells respectively for 24 h,48 h and 72 h. The expression of P-gp was determined by western blot assay and the activity of P-gp was determined by Rhodamine-123 assay. Results:P-gp protein ex-pression levels were significantly increased by scutelarin. After the incubation for 24 h with scutellarin,P-gp protein expression was up-regulated 2. 34-,2. 65-and 2. 00-fold in Caco-2 cells. After the incubation with scutellarin for 48 h,P-gp protein expression was up-regulated 2. 70-,4. 66-and 3. 13-fold. After the incubation with scutellarin for 72 h,P-gp protein expression was up-regulated 2. 82-, 2. 62-and 1. 84-fold. The intracellular accumulation of rhodamine-123 was significantly decreased by scutellarin,indicating that the ef-flux transport activity of P-gp was increased by scutellarin in Caco-2 cells. Conclusion:Scutellarin can significantly up-regulate P-gp protein expression and increase the efflux transport activity of P-gp in Caco-2 cells.