AIM:To investigate the inhibitory ef-fect and mechanism of hesperetin(HST)on human gefitinib-resistant NCI-H1975 lung adenocarcinoma cells.METHODS:CCK-8 assay was used to detect the effects of HST on the proliferation of NCI-H1975 cells;Annexin V-FITC/PI double staining was used to detect HST-induced apoptosis of NCI-H1975 cells;flow cytometry was used to observe the effects of HST and HST+acetylcysteine(NAC)combined on the levels of reactive oxygen species(ROS)in NCI-H1975 cells;Western blot was used to detect the expression of Bcl-2,Bax,Cleaved Cas-pase-3,p-eIF2α,eIF2α and CHOP proteins in NCI-H1975 cells by HST,NAC+HST and Salubrinal+HST.The antitumor effect of HST in vivo was stud-ied by constructing a xenograft model in nude mice;HE staining was used to observe the effect of HST on the histopathological morphology of heart,liver,kidney and xenograft in tumor-bearing mice;immunohistochemistry was used to detect the ef-fect of HST on p-eIF2α protein in xenograft tissues.RESULTS:Compared with the control group,HST at 37.5 μmol/L for 24 h significantly inhibited the via-bility of NCI-H1975 cells(P＜0.05),and NAC attenu-ated the inhibitory effect of HST;concentrations greater than 150 μmol/L increased intracellular ROS levels(P＜0.05),induced apoptosis(P＜0.05),and increased Caspase3 activity(P＜0.01),and com-pared with HST 300 μmol/L group,ROS levels,apoptosis rate,and Caspase3 activity were signifi-cantly decreased in NAC+HST 300 μmol/L group(P＜0.01);HST up-regulated Bax,Cleaved Caspase-3,CHOP,and p-elF2α expression and down-regulated Bcl-2 expression in a concentration-dependent manner(P＜0.01),and compared with HST 300μmol/L group,Bax and Cleaved Caspase-3 expres-sion was decreased and Bcl-2 expression was in-creased in Sal+HST 300 μmol/L group;p-eIF2αand CHOP expression were significantly down-regu-lated in the NAC+HST 300 μmol/L and Sal+HST 300 μmol/L groups(P＜0.01).In vivo,experiments showed that HST could significantly inhibit the growth of transplanted tumors and up-regulate p-eIF2α protein expression(P＜0.05),and had no sig-nificant adverse effects on the growth status,body weight and important organs(heart,liver and kid-ney)of nude mice.CONCLUSION:HST inhibits the proliferation of gefitinib-resistant NCI-H1975 lung adenocarcinoma cells in vitro and in vivo,and the mechanism may be related to HST mediating ER stress-induced apoptosis of NCI-H1975 cells through ROS.

AIM:To investigate the inhibitory ef-fect and mechanism of hesperetin(HST)on human gefitinib-resistant NCI-H1975 lung adenocarcinoma cells.METHODS:CCK-8 assay was used to detect the effects of HST on the proliferation of NCI-H1975 cells;Annexin V-FITC/PI double staining was used to detect HST-induced apoptosis of NCI-H1975 cells;flow cytometry was used to observe the effects of HST and HST+acetylcysteine(NAC)combined on the levels of reactive oxygen species(ROS)in NCI-H1975 cells;Western blot was used to detect the expression of Bcl-2,Bax,Cleaved Cas-pase-3,p-eIF2α,eIF2α and CHOP proteins in NCI-H1975 cells by HST,NAC+HST and Salubrinal+HST.The antitumor effect of HST in vivo was stud-ied by constructing a xenograft model in nude mice;HE staining was used to observe the effect of HST on the histopathological morphology of heart,liver,kidney and xenograft in tumor-bearing mice;immunohistochemistry was used to detect the ef-fect of HST on p-eIF2α protein in xenograft tissues.RESULTS:Compared with the control group,HST at 37.5 μmol/L for 24 h significantly inhibited the via-bility of NCI-H1975 cells(P＜0.05),and NAC attenu-ated the inhibitory effect of HST;concentrations greater than 150 μmol/L increased intracellular ROS levels(P＜0.05),induced apoptosis(P＜0.05),and increased Caspase3 activity(P＜0.01),and com-pared with HST 300 μmol/L group,ROS levels,apoptosis rate,and Caspase3 activity were signifi-cantly decreased in NAC+HST 300 μmol/L group(P＜0.01);HST up-regulated Bax,Cleaved Caspase-3,CHOP,and p-elF2α expression and down-regulated Bcl-2 expression in a concentration-dependent manner(P＜0.01),and compared with HST 300μmol/L group,Bax and Cleaved Caspase-3 expres-sion was decreased and Bcl-2 expression was in-creased in Sal+HST 300 μmol/L group;p-eIF2αand CHOP expression were significantly down-regu-lated in the NAC+HST 300 μmol/L and Sal+HST 300 μmol/L groups(P＜0.01).In vivo,experiments showed that HST could significantly inhibit the growth of transplanted tumors and up-regulate p-eIF2α protein expression(P＜0.05),and had no sig-nificant adverse effects on the growth status,body weight and important organs(heart,liver and kid-ney)of nude mice.CONCLUSION:HST inhibits the proliferation of gefitinib-resistant NCI-H1975 lung adenocarcinoma cells in vitro and in vivo,and the mechanism may be related to HST mediating ER stress-induced apoptosis of NCI-H1975 cells through ROS.

AIM: To study the effect and mechanism of Delicaflavone on migration and invasion of gefitinib-resistant lung cancer cell line PC-9/GR. METHODS: MTT assay was used to detect cell viability. Transwell and scratch assays were used to detect cell invasion and migration abilities. Western blotting was used to detect the expressions of MMP-9, MMP-2, E-cadherin, N-cadherin, Vimentin and PI3K/Akt/mTOR pathway-related proteins in PC-9/GR cells. RESULTS: Compared with control group, 20 mg/L Delicaflavone could significantly inhibit the viability of PC-9/GR cells for 24 h (P<0.05), while Delicaflavone below 10 mg/L had no significant effect on cell proliferation. The number of invasive cells and migrated cells were decreased significantly by Delicaflavone in a concentration-dependent way (P<0.05 and P<0.01). Delicaflavone could concentration-dependently reduce the expression of MMP-9, MMP-2, N-cadherin, vimentin (P<0.01), meanwhile up-regulate the expression of E-cadherin (P<0.01). In addition, Delicaflavone also decreased the expression of p-PI3K, p-Akt and p-mTOR in a concentration-dependent manner (P<0.01). CONCLUSION: Delicaflavone can inhibit the migration and invasion of PC-9/GR cells by regulating epithelial-mesenchymal transition via PI3K/Akt/mTOR pathway.

Objective To observe the effect of autophagy on paclitaxel-induced CaSki cell death through the regulation of the expression of autophagy gene Beclin1, and to explore the interaction and relationship between autophagy and apoptosis. Methods Eukaryotic expression vector pcDNA3.1-Beclin1 and RNA interference vector pSUPER-Beclin1 were transfected into human cervical cancer CaSki cells in vitro and screened for stable expression cell lines. The formation of autophagic vacuoles was observed with an electronic microscope. The expression of Beclin1 and LC3 was measured by Western blot. After being treated with paclitaxel, the change of cell proliferation was assessed by MTT assay, the percentage of apoptotic cells and autophagic cells were analyzed by flow cytometry. Results A lot of autophagic vacuoles were observed in pcDNA3.1-Beclin1 cells by electronic microscopy. Beclin1 and LC3 protein expression was up-regulated in CaSki cells transfected with pcDNA3.1-Beclin1, and was inhibited in cells transfected with pSUPER-Beclin1. MTT assay revealed the survival rate of CaSki cells was significantly decreased after being transfected with pcDNA3.1-Beclin1. After being treated with paclitaxel, the percentages of apoptotic cells and autophagic cells were both increased in pcDNA3.1-Beclin1 group compared with that of the blank control group especially the increase of apoptosis was particularly evident. Conclusion Autophagy and apoptosis have different roles in the process of paclitaxel-induced cervical cancer CaSki cell line death. Overexpression of Beclin1 in CaSki cells may enhance the apoptosis induced by paclitaxel.

OBJECTIVE:To discuss clinical pharmacists' role in clinical drug treatment. METHODS: Case study was performed retrospectively on clinical pharmacists' involving in clinical drug treatment. RESULTS & CONCLUSION: Hospital pharmacists' participating in drug treatment had achieved remarkable outcome. Clinical pharmacists have to cooperate with medical staff in clinical drug treatment so as to bring their due role into full play.

OBJECTIVE:To investigate the cognition of medical staff's cognition on clinical pharmaceutical care.METHODS:A total of 105 questionnaires collected from the doctors and nurses were subjected to an aggregate analysis in terms of their ultimate educational background,professional title,working lifetime,cognition on clinical pharmaceutical care etc.RESULTS & CONCLUSIONS:Doctors and nurses had positive attitude toward clinical pharmacy care and it is greatly demanded in drug use.Clinical pharmaceutical staff should keep improving their expertise level and strengthen cooperation with medical staff so as to facilitate the development of clinical pharmaceutical care.