ABSTRACT
OBJECTIVE: To investigate the effect of ABT-263,an anti-apoptotic protein inhibitor,on human cutaneous squamous cell carcinoma A431 cells,and to explore its molecular mechanisms. METHODS: i) Total protein was extracted from human immortalized epidermal cells( Ha Ca T cells) and A431 cells in logarithmic growth phase. The protein expression of B-cell lymphoma-2( BCL-2) and BCL2-like 1( BCL-XL) was detected by Western blotting. ii) The A431 cells were treated with ABT-263( inhibitor group) and dimethyl sulfoxide( control group) at a concentration of 50 μmol/L for 4 and 9 hours. The morphological changes of the cells were examined by transmission electron microscopy. iii) The A431 cells were treated with 0,10,25,40,and 50 μmol/L of ABT-263 for 24 hours,and the cell viability was determined by CCK-8 assay. iv) The A431 cells were treated with different doses of ABT-263,and the expression of cleaved Caspase-3, cleaved poly( ADP-ribose) polymerase-1( PARP-1), phosphorylated protein kinase B [p AKT(ser473)],phosphorylated glycogen synthase kinase-3β(p GSK3β) and phosphorylated histone H2 AX(γH2 AX) was detected by Western blot. RESULTS: The relative expression of BCL-2 and BCL-XL in A431 cells were higher than those in Ha Ca T cells( P < 0. 01). Transmission electron microscopy results showed that A431 cells in inhibitor group gradually changed from normal morphology to apoptotic morphology,showing loss of microvilli,increased nuclear chromatin density and aggregation around the nuclear membrane,and nuclear fragmentation. The cell viability of A431 cells in 10,25,40 and 50 μmol/L groups were lower than those in control group( P < 0. 05). The relative expression of cleaved Caspase-3 and cleaved PARP-1 in A431 cells in 10,30 and 50 μmol/L groups were higher than those in control group( P < 0. 05).The relative expression of p AKT( ser473) and p GSK3β in A431 cells in 10,25,40 and 50 μmol/L groups were lower than those of the control group( P < 0. 05) and γH2 AX protein expression was higher than that of the control group( P <0. 05). A431 cell viability and p GSK3β protein expression decreased with the increase of inhibitor dosage( P < 0. 01).The relative expression of cleaved Caspase-3 and γH2 AX protein increased with the increase of inhibitor dosage( P <0. 01),showing dose-effect relationship. CONCLUSION: ABT-263 can induce apoptosis of A431 cells through mitochondria pathway and induce the inactivation of AKT/GSK3β pathway,which can promote the apoptosis of A431 cells with a doseeffect relationship.
ABSTRACT
Objective: To compare the different therapeutic effects of acupuncture and Phenolphthalein for constipation due to intestinal qi stagnation. Methods:A total of 50 patients with constipation due to intestinal qi stagnation were randomly divided into an acupuncture group and a medication group by the random digital table, 25 cases in each group. The patients in the acupuncture group were treated by puncturing Gongsun (SP 4), Sanyinjiao (SP 6), Taichong (LR 3), Zusanli (ST 36), Shangjuxu (ST 37), Hegu (LI 4), Lieque (LU 7), and Tianshu (ST 25), once every day, and 7 d as one course, for continuous 3 courses; while the patients in the medication group were given Phenolphthalein, 7 d as one course, for continuous 3 courses. Compared the Cleveland clinic constipation score (CCS) between the two groups after one course, 3 courses and 3 months after the treatment, as well as the frequency of defecation within one week. Results:After one week of treatment, CCS scores and frequency of defecation per week were significantly changed in both groups compared with those before treatment (P<0.05), and CCS scores and frequency of defecation per week were improved more significantly in the patients of the medication group than in those of the acupuncture group (P<0.01). After 3 weeks of treatment, CCS scores and frequency of defecation per week were significantly changed in both groups compared with those before treatment (P<0.05), and CCS scores and frequency of defecation per week were improved more significant in the patients of the acupuncture group than in those of the medication group (P<0.05). Three months after the end of treatment, CCS scores and frequency of defecation per week were significantly changed in both groups compared with those before treatment (P<0.05), and CCS scores and frequency of defecation per week were improved more significantly in the patients of the acupuncture group than in those of the medication group (P<0.05). Conclusion:Acupuncture and medication are effective in the treatment of constipation due to intestinal qi stagnation. Their short-term therapeutic effect is similar, but the long-term therapeutic effect is better in the acupuncture group than in the medication group.
ABSTRACT
Objective:To compare the therapeutic effects between the Governor Vessel-regulating, liver-soothing and mind-calming needling and the routine acupuncture therapy for insomnia due to liver fire disturbing the heart. Methods:A total of50 cases with insomnia were randomly divided by the random numberl table into the two groups, 25 cases in each group. The observation group was treated with the Governor Vessel-regulating, liver-soothing and mind-calming needling, while the control group was treated with the routine acupuncture therapy. The two groups were treated for five sessions per week, ten sessions as a course. After 3-course treatment, the therapeutic effects and the improvement of Pittsburgh sleep quality index (PSQI) were observed. Results:After treatment for three courses, the total effective rate was 100% in the treatment group and 92% in the control group. The difference in the therapeutic effect between the two groups was statistically significant (P0.05). Conclusion:Thetherapeutic effect of Governor Vessel-regulating, liver-soothing and mind-calming needling is better for insomnia due to liver fire disturbing the heart than the routine acupuncture therapy.