ABSTRACT
The most common problem in the diagnosis of gastrointestinal stromal tumor (GIST) is inadequate specimen fixation. The paper focused on specimen fixation and standardized protocol in immunohistochemistry staining and gene mutation detection. We have adjusted some procedure used in immunohistochemistry staining and c-kit gene detection to improve the quality of inadequately fixed specimen. It maybe useful for clinicians, pathologists and technicians working in immunohistochemistry labs and gene detection labs.
Subject(s)
Humans , Gastrointestinal Stromal Tumors , Diagnosis , Pathology , Immunohistochemistry , Mutation , Proto-Oncogene Proteins c-kit , Genetics , Specimen Handling , Staining and LabelingABSTRACT
<p><b>OBJECTIVE</b>To investigate the clinicopathological value of the expression and amplification of P21-activated kinase 1 gene (PAK1) in colorectal carcinoma(CRC).</p><p><b>METHODS</b>Immunohistochemistry (IHC), fluorescence in situ hybridization (FISH) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling(TUNEL) methods were used to examine the protein expression, amplification of PAK1 and cell apoptosis in 80 cases of CRC and 30 cases of colorectal adenoma by tissue microarray.</p><p><b>RESULTS</b>IHC showed an overexpression of PAK1 protein in 26% of colorectal adenomas and 62% of CRCs. Significant association was found between expression of PAK1 and tumor histological grade as well as tumor clinical stage(P<0.05). In poor-differentiated(G(3)) CRCs, PAK1 expression in 90% carcinoma was up-regulated, which was significantly higher than that in tumors of G(1/2)(51%). Overexpression of PAK1 was detected in 78% of CRCs in later clinical stages (Dukes C, D), which was significantly higher than that in early clinical stages (Dukes A,B, 53%). In addition, negative correlation between PAK1 overexpression and cell apoptosis was observed in these CRC cohorts(P<0.05). FISH revealed that amplification of PAK1 gene was examined in only 3% CRCs.</p><p><b>CONCLUSIONS</b>Overexpression of PAK1 protein may play an important role in development and progression of colorectal neoplasms and it is closely associated with the malignant histological and invasive phenotype of CRCs. The expression of PAK1 in CRC may be used as one of the new molecular markers in predicting tumors malignant potential and progression.</p>
Subject(s)
Female , Humans , Male , Middle Aged , Apoptosis , Colorectal Neoplasms , Genetics , Pathology , Gene Expression , Gene Expression Regulation, Neoplastic , Immunohistochemistry , In Situ Hybridization, Fluorescence , Neoplasm Staging , p21-Activated Kinases , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To research the treatment effect of complex prescription of Chinese crude drug in BALB/c athymic mice with human liver cancer, which were built by Bel-7402.</p><p><b>METHOD</b>48 male BALB/c athymic mouse models were built by Bel-7402 with an indirect method. After 24 hours of postoperation, the 48 athymic mice were distributed randomly into 4 groups which were treated by intragastric administration with complex prescription of Chinese crude drug that had been deliquated into 3 groups by the different density as the low, middle, and high and FT207 (Tegafur) for 4 weeks. At last, athymic mice were put to death and PTEN was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry (PowerVision Two-Step Histostaining Reagent).</p><p><b>RESULT</b>All of the 48 athymic mice survived 12 to 28 days (Ms 24 days) and every mouse with liver cancer demonstrated by dissection. The result of immunohistochemistry represents that the intensity of PTEN in latero-cancer tissue is the highest, and then the hepatic tissue, the lowest is cancer tissue, P < 0.01. It also represents that the intensity of PTEN in treatment groups (A, B, C) is more higher than the control group (D), P < 0.05 or P < 0.01, and group B is the highest in the treatment groups, P < 0.05 or P < 0.01. However, there is no significant statistic difference between group A and group C.</p><p><b>CONCLUSION</b>The higher expression of PTEN in the laterocancer tissue can represent the protective reaction of stress of the organism. And anticancer effect of this complex prescription of Chinese crude drug relates to an eligible density of it. Mechanisms of this complex prescription of Chinese crude drug healing HCC may partially be explained by enhancing the expression of PTEN in liver.</p>
Subject(s)
Animals , Humans , Male , Mice , Antineoplastic Agents, Phytogenic , Pharmacology , Carcinoma, Hepatocellular , Drug Therapy , Pathology , Cell Line, Tumor , Drug Combinations , Drugs, Chinese Herbal , Pharmacology , Immunohistochemistry , Liver , Pathology , Liver Neoplasms , Drug Therapy , Pathology , Mice, Inbred BALB C , Mice, Nude , PTEN Phosphohydrolase , Metabolism , Phytotherapy , Plants, Medicinal , Chemistry , Random Allocation , Xenograft Model Antitumor AssaysABSTRACT
Hydrophobic interaction chromatography was used to separate correctly refolded and mis-refolded consensus interferon. The effects of ligand types, salt concentration, pH and flow rate were investigated. The best result could be obtained by using Butyl Sepharose 4 Fast Flow, 0.8 mol/L of ammonium sulfate, pH 8.3 and 90cm/h of linear flow rate. Reverse-phase HPLC analysis showed the purity of the pooled fraction was as high as 99.6%. The specific activity of purified consensus interferon was 2.3 x 10(9) IU/mg, The mass recovery of targeth protein was 36.7%.