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Objective To localize endogenous alkaline phesphatase in human neutrophils with ELF(enzyme-labeled fluorescence)-97 phosphate,which yields an intensely fluorescent yellow-green precipitate at the site of enzymatic activity. Methods Neutrophils were isolated from human blood and fixed,then histochemistry with the ELF-97 phosphate was performed with the ELF-97 endogenous phosphatase detection kit.All photography was performed with a Nikon labophot fluorescence/DIC microscope. Results Fluorescent yellow-green granules,well-distributed but size-varied,were observed in neutrophils under fluorescence microscope.There were 94.102?3.133 percent rate of positive neutrophil alkaline phosphatase among 51 cases of healthy volunteers.Conclusion ELF-97 endogenous phosphatase detection kit can provide sensitive,high-resolution localization of endogenous phosphatase activity in neutrophils.
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Objective: To investigate the relationship between the Smad4-independent pathway of TGF-?1 and drug-resistance of pancreatic cancer. Methods: The sensitivities of Smad4 homozygous-deleted pancreatic cancer cell line BxPC3 to different kinds of anti-cancer drugs (5-Fu, Gemzar, Oxaliplatin, Cisplatin, CPT-11 and Epirubicin) were observed by MTT assay before and after they were transfected with full-length cDNA of TGF-?1 or treated with TGF-?1 (5 and 10 ng/ml) solution. Western blot was used to detect p170 protein expression after stimulation with different concentrations of TGF-?1. Results: Cisplatin had the most powerful killing effect on BxPC3 cells, followed by Oxaliplatin, 5-Fu and CPT-11 with moderate effect and Gemzar and Epirubicin with the least effect. Cells transfected with full-length cDNA of TGF-?1 or treated with TGF-?1 solution became less sensitive to Cisplatin. Western blot revealed upregulation of p170 expression by TGF-?1. Conclusion: The Smad4-independent pathway of TGF-?1 can increase the drug resistance of pancreatic cancer cells through upregulating expression of p170.
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Objective:To investigate the expression of PKC? and TGF-? proteins in exocrine pancreatic carcinoma and their relationship. Methods: Forty-two specimens of pancreatic carcinoma and their adjacent tissues were examined for PKC? and TGF-? expression by tissue microarray technique and immunohistochemistry. Results: The expressions of TGF-?1 in the pancreatic carcinoma tissues and their adjacent tissue were 80. 9%(34/42) and 19. 1%(8/42), respectively(P
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Objective To observe the age-related changes of brain-derived neurotrophic factor(BDNF) and investigate the effects of ginsenosides(GS) on BDNF in nucleus basalis of Meynert(NBM) and cerebral cortex of aged rats. Methods Twenty-four female Wistar rats were randomly divided into 3 groups: young group(3-5 months),aged groups(27 months) and GS group(27 months).GS group was fed with GS from 18 to 27 months.Immunohistochemistry and Imaging analysis were used to show the expression and distribution of BDNF in NBM and cerebral cortex of each group. Results BDNF level of aged group was much lower than that of young group in these two brain areas(P