OBJECTIVE To investigate the inhibitory effects of Bruceine A(BA)on colon cancer and its underlying mechanisms.METHODS Human colon cancer HT-29 and HCT116 cells were treated with various concentrations of BA(0,1,2,5,10,20,40,80 μmol·L-1).Cell viability was assessed using the Cell Counting Kit-8(CCK-8).Flow cytometry,wound healing assays,and Transwell assays were employed to evaluate the effects of BA on cell apoptosis,cell cycle,invasion,and migration.Mo-lecular docking simulations were used to assess the binding of BA to GSK-3β protein,and Western blot analysis was used to examine protein expression related to the cell cycle,epithelial-mesenchymal transition,and the Wnt/β-catenin signaling pathway.An HT-29 cell subcutaneous xenograft mouse model was established.After tumor formation,mice were randomly divided into three groups(six mice per group):a blank group,a low-dose BA group(0.1 mg·kg-1),and a high-dose BA group(0.2 mg·kg-1).Mice were ad-ministered the drug for 19 d,then sacrificed,and tumor tissues were collected.Tumor volume changes over time were observed;Ki67 immunohistochemistry was used to assess cell proliferation in tumor tissues;Western blot analysis of Wnt/β-catenin signaling pathway protein expression was conducted.RESULTS Compared with the blank group,BA could significantly inhibit the proliferation of HT-29 and HCT116 cells,with IC50 values of 10.80 μmol·L-1 and 17.96 μmol·L-1,respectively.Flow cytometry results showed that BA significantly induced apoptosis of HT-29 cells(P<0.01,P<0.001),and arrested the cell cycle at the S phase,accompanied by de-creased expression of cycle-related proteins CDK2 and Cyclin A(P<0.05,P<0.01,P<0.001).BA inhibited cell migration and in-vasion ability(P<0.05,P<0.01,P<0.001),reduced the expression of EMT-related proteins Snail,Vimentin,and N-Cadherin(P<0.01,P<0.001),and upregulated the expression of E-Cadherin protein.In addition,BA inhibited the expression of β-catenin and p-GSK3β proteins.Wnt agonist LiCl could significantly antagonize the anti-colon cancer effect of BA;Wnt inhibitor XAV939 could enhance the anti-colon cancer effect of BA.In the in vivo experiment,compared with the blank group,the tumor volume of the low-dose and high-dose BA groups was significantly reduced(P<0.05,P<0.001).Immunohistochemistry results showed that compared with the blank group,the expression of Ki67 in tumor tissues of the low-dose and high-dose BA groups was significantly reduced(P<0.001).Western blot results further proved that BA inhibited the Wnt/β-catenin signaling pathway.CONCLUSION BA inhibits the viability,invasion,and migration of colon cancer HT-29 cells,induces apoptosis,and causes cell cycle arrest.Additionally,it significantly suppresses the growth of subcutaneous HT-29 cell xenografts in vivo,possibly related to the Wnt/β-catenin signaling pathway.

OBJECTIVE To investigate the inhibitory effects of Bruceine A(BA)on colon cancer and its underlying mechanisms.METHODS Human colon cancer HT-29 and HCT116 cells were treated with various concentrations of BA(0,1,2,5,10,20,40,80 μmol·L-1).Cell viability was assessed using the Cell Counting Kit-8(CCK-8).Flow cytometry,wound healing assays,and Transwell assays were employed to evaluate the effects of BA on cell apoptosis,cell cycle,invasion,and migration.Mo-lecular docking simulations were used to assess the binding of BA to GSK-3β protein,and Western blot analysis was used to examine protein expression related to the cell cycle,epithelial-mesenchymal transition,and the Wnt/β-catenin signaling pathway.An HT-29 cell subcutaneous xenograft mouse model was established.After tumor formation,mice were randomly divided into three groups(six mice per group):a blank group,a low-dose BA group(0.1 mg·kg-1),and a high-dose BA group(0.2 mg·kg-1).Mice were ad-ministered the drug for 19 d,then sacrificed,and tumor tissues were collected.Tumor volume changes over time were observed;Ki67 immunohistochemistry was used to assess cell proliferation in tumor tissues;Western blot analysis of Wnt/β-catenin signaling pathway protein expression was conducted.RESULTS Compared with the blank group,BA could significantly inhibit the proliferation of HT-29 and HCT116 cells,with IC50 values of 10.80 μmol·L-1 and 17.96 μmol·L-1,respectively.Flow cytometry results showed that BA significantly induced apoptosis of HT-29 cells(P<0.01,P<0.001),and arrested the cell cycle at the S phase,accompanied by de-creased expression of cycle-related proteins CDK2 and Cyclin A(P<0.05,P<0.01,P<0.001).BA inhibited cell migration and in-vasion ability(P<0.05,P<0.01,P<0.001),reduced the expression of EMT-related proteins Snail,Vimentin,and N-Cadherin(P<0.01,P<0.001),and upregulated the expression of E-Cadherin protein.In addition,BA inhibited the expression of β-catenin and p-GSK3β proteins.Wnt agonist LiCl could significantly antagonize the anti-colon cancer effect of BA;Wnt inhibitor XAV939 could enhance the anti-colon cancer effect of BA.In the in vivo experiment,compared with the blank group,the tumor volume of the low-dose and high-dose BA groups was significantly reduced(P<0.05,P<0.001).Immunohistochemistry results showed that compared with the blank group,the expression of Ki67 in tumor tissues of the low-dose and high-dose BA groups was significantly reduced(P<0.001).Western blot results further proved that BA inhibited the Wnt/β-catenin signaling pathway.CONCLUSION BA inhibits the viability,invasion,and migration of colon cancer HT-29 cells,induces apoptosis,and causes cell cycle arrest.Additionally,it significantly suppresses the growth of subcutaneous HT-29 cell xenografts in vivo,possibly related to the Wnt/β-catenin signaling pathway.

Objective:To compare the short-term curative effects between 3 fixations of the tibial end with an interface screw, an interface screw + a portal nail and an interface screw + a knotless rivet in reconstruction of the anterior cruciate ligament (ACL).Methods:A retrospective study was conducted to analyze the data of 150 patients who had undergone reconstruction surgery for ACL injury at Department of Orthopaedics, Jinling Clinical Medical College, Nanjing Medical University from January 2019 to December 2021. The patients were divided into 3 groups according to their different fixations of the tibial end. In group A of 51 cases subjected to fixation of the tibial end with an interface screw, there were 40 males and 11 females with a mean age of (29.8±10.6) years. In group B of 55 cases whose tibial end was fixated with an interface screw + a portal nail, there were 47 males and 8 females with a mean age of (28.6±6.9) years. In group C of 44 cases whose tibial end was fixated with an interface screw + a knotless rivet, there were 39 males and 5 females with a mean age of (28.6±8.1) years. The 3 groups were compared in terms of International Knee Documentation Committee (IKDC) scoring and Lysholm knee function scoring at 12 and 48 weeks after operation, incidence of local pain at 4 weeks after operation, and stability of the knee at 48 weeks after operation.Results:There was no significant difference in the preoperative general data or follow-up time between the 3 groups, indicating comparability ( P>0.05). At 12 weeks after surgery, in Group B and Group C, the IKDC scores [(74.27±3.32) and (76.48±3.83) points] and Lysholm knee joint scores [(77.65±3.14) and (79.75±5.39) points] were significantly better than those in Group A [(69.73±6.04) and (71.63±3.36) points] ( P<0.05). However, there was no statistically significant difference in the above scores among the 3 groups at 48 weeks after surgery ( P>0.05). The incidence of local pain at the tibial end in group B (9.1%, 5/55) was significantly higher than that in group A (0, 0/51) and group C (2.3%, 1/44) at 4 weeks after surgery, and the results of the anterior drawer test in group A (36 negative cases, 14 cases of grade Ⅰ, and 1 case of grade Ⅱ) were significantly worse than those in group B (50 negative cases, and 5 cases of grade Ⅰ) and group C (40 negative cases, and 4 cases of grade Ⅰ) at 48 weeks after surgery ( P<0.05). Conclusions:In ACL reconstruction, all the 3 fixations of the tibial end can achieve fine short-term curative effects. Fixation with an interface screw + a knotless rivet may lead to a lower rate of ligament relaxation and a higher score of early knee function than fixation with an interface screw, and a lower incidence of local pain at the tibial end than fixation with an interface screw + a portal nail.