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.

Neonatal hypoxic-ischemic encephalopathy (HIE)is the brain injury caused by asphyxiation in the perinatal period.HIE is a serious health-threatening disease associated with significant morbidity and mortality,and there is no effective treatment for the disease.Melatonin is a neuroendocrine hormone,which is synthesized primarily by the pineal gland.The synthesis and seretion of melatonin are regulated by light intensity.Melatonin has a high affinity for the central nervous system,which can easily penetrate the blood-brain barrier and blood placental barrier.It has been found that melatonin has functions in regulating the sleep cycle,antioxidant and anti-inflammatory.Melatonin has also been shown to regulate lipid and glucose metabolism.Recent research suggests that the melatonin appears to be a versatile anti-oxidative,anti-inflammatory,anti-apoptotic agent,as well as a molecule with regulation of mitochondrial function and autophagy process,and plays a neuroprotective role in HIE.Therefore,this review focuses on the established mechanisms of injury brain protection,progress of animal studies and clinical trials of melatonin in HIE,to provide references for its clinical application.

Objective To explore the effect of high sn-2 palmitate infant formula (HPIF) on stool frequency and consistency,fatty acids,calcium and magnesium contents in infants.Methods A prospective,double-blind,randomized,controlled clinical study was conducted including 94 healthy mature infants of single birth and appropriate for gestational age,born from June 2013 to December 2014.All eligible infants were enrolled within 21 days after birth.All the infant formula fed subjects were divided randomly into two groups as standard infant formula (IF) group and high sn-2 palmitate infant formula (HPIF) group.Breast-fed infants were enrolled as control group (BF group).All infants were followed up until 90 days old.The growth indexes and defecation status of the three groups were monitored dynamically.Meanwhile,stool fatty acid profile and mineral contents were also detected.Results There was no significant difference in head circumference,body length and body weight among the three groups at enrollment,42 days and 90 days old.The stool frequency and mushy stool frequency of HPIF and IF groups were significantly lower than that of BF group at 42 days and 90 days old;formed stool frequency was higher in HPIF and IF groups than in BF group.The fecal palmitic acid level in dry feces was significantly higher in HPIF and IF groups than in BF group [(31.1 ± 9.8),(30.9± 10.7) vs.(10.8± 8.8) mg/g] at 42 days old.At 90 days old,the fecal palmitic acid level in dry feces was significantly lower in HPIF group than in IF group [(24.3± 9.8) vs.(29.9± 7.9) mg/mg],while was significantly higher in both infant formula fed groups than in BF group [(8.9± 8.4) mg/g].The fecal calcium level in dry feces of HPIF and IF groups were significantly higher than that of BF group [(38.3± 14.0),(38.8± 15.5) vs.(21.3± 13.7) mg/g] at 42 days old.At 90 days old,the fecal calcium level in dry feces of HPIF group was significantly lower than that of IF group [(31.1 11.2) vs.(45.9 ± 16.5) mg/g,dry stool] and significantly higher than that of BF group [(21.5 ± 9.9) mg/g].The fecal magnesium level was similar between HPIF and IF groups,and significantly higher than that of BF group at 42 days and 90 days old.The fecal calcium level was positively correlated with the content of fecal palmitic acid among three groups (r =0.43,P＜ 0.01).Conclusions Breast milk is the best food for infants.Compared with standard infant formula,feeding with high sn-2 palmitate infant formula can reduce the fecal excretion of calcium and palmitic acid,making it closer to the level of breast-fed infants.