ABSTRACT
OBJECTIVE: To investigate the role of N-methyl-D-aspartate receptor(NMDAR)/cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA) signaling pathways in regulating 2-chloroethanol-induced aquaporin-4(AQP4) expression in astrocytes(AS). METHODS: i) AS in logarithmic growth phase were treated with 2-chloroethanol at the doses of 0.0, 7.5, 15.0 and 30.0 mmol/L for 12 hours, and the cells were collected for detection. ii) The AS in logarithmic growth phase were divided into blank control group, inhibitor control group, 2-chloroethanol group, and inhibitor intervention group. The inhibitor included dizocilpine(MK-801) and N-(2-[p-bromocinnamylamino-]ethyl)-5-isoquinolinesulfonamide(H89). The blank control group did not receive any treatment. The inhibitor control group was treated with a concentration of 10.0 μmmol/L MK-801 or 15.0 μmmol/L H89. The MK-801 intervention group was pretreated with MK-801 at a concentration of 10.0 μmmol/L for 30 minutes. The H89 intervention group was pretreated with H89 at a concentration of 15.0 μmmol/L for 1 hour. After the intervention, the AS in 2-chloroethanol group and MK-801, H89 intervention group were stimulated with 2-chloroethanol at a dose of 30.0 mmol/L for 12 hours. iii) The AS in each group were collected and used for Western blotting and real-time fluorescence quantitative polymerase chain reaction analysis to detect the protein and mRNA expression of AQP4, NMDAR receptor main subunit(NR1), NMDAR receptor 2 B subunit(NR2 B) and calmodulin dependent protein kinaseⅡ(CaMKⅡ). The Western blotting was adopted to detect the expression of phosphorylase-CaMKⅡ(p-CaMKⅡ) and PKA. Colorimetric method was used to detect the concentration of calcium(Ca~(2+)) in AS. The enzyme-linked adsorption test was used to measure adenylate cyclase(AC) activity and cAMP levels. RESULTS: i) The relative expression of protein and mRNA of AQP4, NR1 and NR2 B, PKA at protein level and CaMKⅡ at mRNA level, and the ratio of p-CaMKⅡ/CaMKⅡ protein, the concentration of Ca~(2+), AC activity and cAMP level in 30.0 mmol/L group were higher then those of 0.0 mmol/L group in AS(P<0.05). The relative protein expression of PKA and the concentration of Ca~(2+) increased with the increase of 2-chloroethanol(P<0.05). ii) The relative protein expression of AQP4 and the concentration of Ca~(2+) in the 2-chloroethanol group were higher than that of the blank control group and MK-801 control group(P<0.05). The relative protein expression of AQP4 and the concentration of Ca~(2+) in MK-801 intervention group were lower than that in 2-chloroethanol group(P<0.05). The relative protein expression of AQP4 and PKA in 2-chloroethanol group were higher than that of the blank control group and H89 control group(P<0.05). The relative protein expression of AQP4 and PKA in H89 intervention group was lower than that in 2-chloroethanol group(P<0.05). CONCLUSION: The 2-chloroethanol timulation induces the expression of AQP4 by activating NMDAR/cAMP/PKA signaling pathway in AS.
ABSTRACT
@#Objective To explore the effect of postoperative progressive resistance training (PRT) on the upper limb function in breast cancer patients. Methods 66 breast cancer patients were randomly divided into intervention group (n=33) and control group (n=33). The in-tervention group accepted a 12-week PRT and the control group accepted the routine training since 4-6 weeks postoperative as they admitted to hospital for their first chemotherapy. All the patients received a same content of health education. Results The grip strength, range of mo-tion of abduction and flexion of shoulder were more in the intervention group than in the control group after intervention (P<0.01). Conclu-sion PRT can effectively improve the function of upper limbs in breast cancer patients.
ABSTRACT
Objective:To study the effects of smokeless tobacco extract(ST)on the proliferation and the heat shock protein 70 (HSP70)expression of human periodontal ligament cells(hPDLCs).Methods:hPDLCs were cultured in vitro and identified by im-munohistochemistry(IHC).The cells were stimulated with ST at 0.01 6 -50 g/L respectively for 24 h,the proliferation was examine by MTT assay,HSP70 expression was detected by immunehistochemical staining and Western Blot.Results:ST inhibited the prolifer-ation and increased HSP70 expression in cytoplasm and nucleus at 0.4 -50 g/L dose dependantly.Conclusion:ST may inhibit the proliferation and increase HSP70 expression of hPDLCs in a dose depandant manner.