ABSTRACT
To study the variation of six ester-type alkaloids and characteristic fingerprints in the process from Radix Aconite Lateralis to Heishunpian and lay a foundation for the study of the processing principle of Heishunpian, HPLC. analysis was performed on a Phenomenex Gemini C18 (4.6 mm x 250 mm, 5 microm) with acetonitrile and 40 mmol x L(-1) ammonium acetate (adjusted to pH 10 with concentrated ammonia water) as mobile phase. The detection wavelength was set at 235 nm. The flow rate was set at 0.8 mL x min(-1) and the injection volume was 10-20 microL. Six ester-type alkaloids were determined and characteristic fingerprints of the process were established. As the process continues, the contents of diester diterpene alkaloids were decreased step by step, while the contents varia tion of monoester diterpene alkaloids were not obvious. Each sample showed significant difference in characteristic fingerprints. With the exception of 6 known monoester diterpene alkaloids and diester diterpene alkaloids, 13 peaks were marked in the characteristic fingerprints, of which the total change rule of the other 7 unknown peaks were similar with 3 diester diterpene alkaloids. The established method is accurate, reliable and repeatable, and can provide reference for revealing change rule of index components and illuminating processing principle in the process of Heishunpian.
Subject(s)
Aconitine , Chemistry , Aconitum , Chemistry , Alkaloids , Chemistry , Chemistry, Pharmaceutical , Methods , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Chemistry , Esters , Chemistry , Molecular StructureABSTRACT
<p><b>OBJECTIVE</b>To study the expression and intracellular localization of FRZB gene in gastric cancer tissue, and to explore its significance in gastric cancer.</p><p><b>METHODS</b>The expression of FRZB in tumor tissues from 90 patients with gastric cancer and in normal gastric mucous as control were analyzed by immunohistochemistry in tissue array. FRZB expression in gastric cancer cell lines and immortalized gastric epithelial cell line GES-1 were detected by quantitative real-time PCR(Q-PCR) and Western blot. The intracellular localization of FRZB was observed by immunofluorescence staining.</p><p><b>RESULTS</b>The positive expression rate of FRZB in gastric cancer was 92.2%. FRZB expressed in gastric cancer with well differentiation was higher than that with poor differentiation.The positive rate in normal gastric mucous was 10.0% (one out of ten). By confocal microscope, FRZB localized both in cytoplasma and nucleus, especially on the nuclear membrane. The Q-PCR and Western blot results also showed that the expression of FRZB in gastric cancer cell lines was higher than that in GES-1.</p><p><b>CONCLUSIONS</b>The expression of FRZB in gastric cancer is correlated with tumor cell differentiation and tumor Lauren classification. The nuclear localization of FRZB may contribute to its function in gastric cancer formation and progression.</p>
Subject(s)
Female , Humans , Male , Biomarkers, Tumor , Genetics , Metabolism , Cell Line, Tumor , DNA Primers , Gene Expression , Glycoproteins , Genetics , Metabolism , Neoplasm Staging , Stomach Neoplasms , Genetics , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of cancer-related gene MPS-1 in gastric cancer and to evaluate its significance in clinical diagnosis and therapy.</p><p><b>METHODS</b>The mRNA expression of MPS-1 was determined by polymerase chain reaction after reverse transcription (RT-PCR) in cancer tissues and adjacent non-cancerous tissues from 42 cases with gastric cancer. The expression levels of MPS-1 in 6 gastric cancer cell lines (AGS, MKN-45, SGC 7901, KATO III, N-87 and SNU-1) were also determined by RT-PCR and Western blot.</p><p><b>RESULTS</b>The MPS-1 mRNA was expressed in all tissues and cell lines. The mRNA expression level of MPS-1 in cancer tissues were 1.37+/- 0.87, significantly higher than 0.99+/- 0.67 in adjacent normal gastric mucous tissues (P< 0.01). The expression of MPS-1 was correlated with TNM stage (P< 0.05), but not with age, gender, tumor size and differentiation. The expression level of MPS-1 mRNA in the primary lesions was hig her in the patients with TNM stages III, IV than those with TNM stages I, II. Meanwhile, RT-PCR and Western blot showed the same results that MPS-1 expression was higher in the six gastric cancer cell lines as compared with that in the normal gastric cell line GES-1.</p><p><b>CONCLUSION</b>The high expression of MPS-1 in gastric cancer indicates that MPS-1 might play an important role in gastric carcinogenesis,which may provide a new target in immunotherapy for gastric cancer.</p>