Objective To observe the effect of acupuncture at different time on dorsal root ganglion(DRG)in neuropathic pain(NP)mice.Different regulatory effects of transient receptor potential vanilloid 1(TRPV1)and substance P(SP)in DRG.Methods A total of64 C57BL/6J mice were randomly divided into two intervention groups:ZT8 group and ZT16 group,among which two intervention groups were randomly divided into control group(8 mice),sham-operation group(8 mice),model group(8 mice)and acupuncture group(8 mice).The chronic constriction injury(CCI)model was prepared by sciatic nerve ligation in model group and acupuncture group,and only the sutured wound after sciatic nerve was exposed in sham-operation group.On the 8th day after operation,acupuncture treatment was performed on mice in the acupuncture group.The intervention methods were the same in the two groups,only the acupuncture time was different.The"Yanglingquan"on the surgical side was taken at the acupoint,and manual acupuncture and needle were performed once a day for 15 minutes each time.The mechanical pain and thermal pain thresholds of each group of mice were measured at 3,7,10 and 14 days after modeling to evaluate their pain.The expression of SP and TRPV1 in the L4-6 DRG on the surgical side of mice was observed by immunofluorescence double labeling method,and the SP content in the tissue around the sciatic nerve on the affected side and the skin on the dorsal fibra of the foot was determined by enzymed-linked immunosorbent assay(ELISA).Results ① Acupuncture intervention could improve mechanical and thermal hyperalgesia in CCI mice.And ZT8 time point acupuncture effect is better than ZT16 time point.②Acupuncture can significantly reduce the expression of TRPV1 and SP in DRG of CCI mice,and the effect of acupuncture at ZT8 time point is better than that at ZT16 time point.③Acupuncture can significantly reduce the SP concentration in the peripheral tissue of CCI mice,and the effect of acupuncture at ZT8 time point is better than that at ZT16 time point.④Acupuncture could significantly reduce the SP concentration in the distal skin of the injured nerve in CCI mice,and there was no significant difference between ZT8 and ZT16.Conclusion Acupuncture at different time has different regulation effects on hyperalgesia in CCI mice,and the regulation effect of ZT8 is better than that of ZT16.This may be related to the different regulatory effects of acupuncture at different time on the expression of TRPV1 and SP in DRG of CCI mice and the local SP concentration of injured nerves.

Objective To observe the effect of acupuncture at different time on dorsal root ganglion(DRG)in neuropathic pain(NP)mice.Different regulatory effects of transient receptor potential vanilloid 1(TRPV1)and substance P(SP)in DRG.Methods A total of64 C57BL/6J mice were randomly divided into two intervention groups:ZT8 group and ZT16 group,among which two intervention groups were randomly divided into control group(8 mice),sham-operation group(8 mice),model group(8 mice)and acupuncture group(8 mice).The chronic constriction injury(CCI)model was prepared by sciatic nerve ligation in model group and acupuncture group,and only the sutured wound after sciatic nerve was exposed in sham-operation group.On the 8th day after operation,acupuncture treatment was performed on mice in the acupuncture group.The intervention methods were the same in the two groups,only the acupuncture time was different.The"Yanglingquan"on the surgical side was taken at the acupoint,and manual acupuncture and needle were performed once a day for 15 minutes each time.The mechanical pain and thermal pain thresholds of each group of mice were measured at 3,7,10 and 14 days after modeling to evaluate their pain.The expression of SP and TRPV1 in the L4-6 DRG on the surgical side of mice was observed by immunofluorescence double labeling method,and the SP content in the tissue around the sciatic nerve on the affected side and the skin on the dorsal fibra of the foot was determined by enzymed-linked immunosorbent assay(ELISA).Results ① Acupuncture intervention could improve mechanical and thermal hyperalgesia in CCI mice.And ZT8 time point acupuncture effect is better than ZT16 time point.②Acupuncture can significantly reduce the expression of TRPV1 and SP in DRG of CCI mice,and the effect of acupuncture at ZT8 time point is better than that at ZT16 time point.③Acupuncture can significantly reduce the SP concentration in the peripheral tissue of CCI mice,and the effect of acupuncture at ZT8 time point is better than that at ZT16 time point.④Acupuncture could significantly reduce the SP concentration in the distal skin of the injured nerve in CCI mice,and there was no significant difference between ZT8 and ZT16.Conclusion Acupuncture at different time has different regulation effects on hyperalgesia in CCI mice,and the regulation effect of ZT8 is better than that of ZT16.This may be related to the different regulatory effects of acupuncture at different time on the expression of TRPV1 and SP in DRG of CCI mice and the local SP concentration of injured nerves.

Objective: To investigate the effect of RNA interference (RNAi) targeting AKT1 and PI3K P85 on the proliferation and invasion of breast carcinoma MCF-7 cells. Methods: The recombinant adenovirus expression vector, which contained short hairpin RNA (shRNA) targeting open reading frames of AKT1 and PI3K P85 (rAd5-siAKT1-siPI3K), was transfected into human breast carcinoma MCF-7 cells. AKT1 and PI3K P85 mRNA and protein expressions were detected by real-time PCR and Western blotting analysis. The expressions of PCNA, cyclinD1, and P53 were also detected by Western blotting analysis. The proliferation and apoptosis of MCF-7 cells were measured by MTT, flow cytometry and 2-dementinal and 3-dementional matrigel assay. Results: Recombinant adenovirus vector rAd5-siAKT1-siPI3K dramatically down-regulated AKT1 and PI3K P85 mRNA and protein expressions in MCF-7 cells; the downstream factors PCNA and cyclin D1 were also down-regulated, while P53 was up-regulated. Growth of MCF-7 cells was inhibited by over 50% in rAd5-siAKT1-siPI3K group as measured by MTT assay, and cell cycle was arrested in G_1/G_0 phase compared with untransfected and rAd5-siCtrl transfected groups. Cell growth on matrigel matrix showed normal cell shapes, while the cells in rAd5-siAKT1-siPI3K transfected group were detached from the matrix or grew in scattered clustering patterns, forming only small aggregates. Conclusion: shRNA targeting AKT1 and PI3K P85 can significantly down-regulate the expression of AKT1 and PI3K P85 in breast carcinoma MCF-7 cells, and inhibit the growth of MCF-7 cells in vitro.

Objective: To study the effect of and possible mechanism of knockinng down PI3Kp85α using siRNA in MCF-7 human breast cancer cell line. Methods: Oligofectamine was used to transfect PI3Kp85α siRNA to knock down the PI3Kp85α expression level in MCF-7 human breast cancer cell line in vitro. Real-time PCR was conducted to detect the expression of PI3Kp85α. The effect of PI3Kp85αsiRNA on the growth of MCF-7 cells was measured by MTT. The cell cycle distribution and cell apoptosis were detected by cell flow cytometry. Protein expression was evaluated by immunofluorescence staining and Western blot. Results: The expression of PI3Kp85 α was knocked down with PI3Kp85α siRNA in MCF-7 cells. Cell growth was delayed in PI3Kp85αsiRNA-treated group. Conclusion: The suppressive effect of PI3Kp85αsiRNA on the growth of MCF-7 human breast cancer cell line is significant and PI3Kp85α could be a candidate for gene therapy for breast cancer.

With the wide application of triazophos in agricultural production, increasing attention has been paid to the contamination of this pesticide to the environment. The studies on triazophos residue and degradation have made certain progress in recent years.This paper gives an outline of micro-organisms degrading triazophos and the possible pathways by which they degrade this pesticide. The reaction kinetics and mechanisms of triazophos hydrolysis and photolysis are also reviewed briefly.