ABSTRACT
<p><b>OBJECTIVE</b>To study the expression of Notch receptors, ligands and downstream target genes in hypertrophic scar and normal skin, and to investigate its role in the development of hypertrophic scar.</p><p><b>METHODS</b>By immunohistochemistry, the expression of epidermal differentiation markers- beta1 integrin, keratin 14 (K14) and keratin 19 (K19), as well as Notch 1-4 and Jagged1 were examined in hypertrophic scars and normal skins. The expression of Notch downstream genes- P21 and P63 was analyzed with real-time quantitative PCR and immunohistochemistry staining.</p><p><b>RESULTS</b>Histological analysis revealed a significant epidermal thickening in the hypertrophic scars, with excessive cell layers above the basal layer. Compared to the normal epidermis, the expression of beta1 integrin, K19 and K14 decreased in hypertrophic scars (P <0.05). Positive expression rate of Notch1 and Jagged1 in keratinocytes was significantly higher in hypertrophic scar than in normal skin (P < 0.05), while there was no difference in Notch2 and 3 positive expression rate. Furthermore, the expression of P21 was significantly up-regulated, while the expression of P63 was down-regulated in keratinocytes of hypertrophic scar (P < 0.05).</p><p><b>CONCLUSIONS</b>Notch signal may play an important role in hypertrophic scar pathogenesis. Over-differentiation of Keratinocytes in hypertrophic scar may be related to the overexpression of Notch1 and Jagged1, up-regulation of P21 gene and down-regulation of P63 gene.</p>
Subject(s)
Adult , Female , Humans , Male , Young Adult , Calcium-Binding Proteins , Metabolism , Case-Control Studies , Cicatrix, Hypertrophic , Metabolism , Pathology , Down-Regulation , Epidermis , Metabolism , Pathology , Integrin beta1 , Metabolism , Intercellular Signaling Peptides and Proteins , Metabolism , Jagged-1 Protein , Keratin-14 , Metabolism , Keratin-19 , Metabolism , Membrane Proteins , Metabolism , Receptor, Notch1 , Metabolism , Serrate-Jagged Proteins , Signal Transduction , Up-RegulationABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of METH1 gene transfection on fibroblast proliferation and I, III collagen synthesis in rabbit ear scar.</p><p><b>METHODS</b>The hypertrophic scar model on the rabbit ears was reproduced. 10 days after epithelization, Ad-METH1 was injected into the scar tissue. 30 days later, the effect of METH1 gene transfection on the angiogenesis, fibroblast proliferation and the ratio of collagen I/III in the scar tissue was detected by microcirculation microscope, AgNOR particle count and collagen dyeing.</p><p><b>RESULTS</b>30 days after injection of Ad-METH1, angiogenesis, fibroblast proliferation and the ratio of collagen I/III in the scar tissue were obviously suppressed.</p><p><b>CONCLUSION</b>Early application of Ad-METH1 after epithelization can markedly inhibit the formation of the hypertrophic scar.</p>
Subject(s)
Animals , Female , Male , Rabbits , ADAM Proteins , Genetics , Angiogenesis Inhibitors , Genetics , Cicatrix, Hypertrophic , Genetics , Pathology , Disease Models, Animal , Ear , Pathology , Ear, External , Pathology , Microcirculation , Neovascularization, Pathologic , Transfection , Wound HealingABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of curcumin (Cur) and erythromycin (EM) on multidrug resistance (MDR) reversal of K562/A02 cell line and their mechanism.</p><p><b>METHODS</b>MTT assay was employed to determine the sensitivity of Cur, EM-treated K562/A02 cells to adriamycin (ADM). Flow cytometry was used to measure intracellular mean fluorescence intensity (MFI) of daunorubicin (DNR). P-gp expression was determined by immunohistochemistry. RT-PCR technique was used to examine the mdr1 mRNA level.</p><p><b>RESULTS</b>IC(50) of ADM in K562/A02 cells was decreased when treated with Cur or EM, and the reversal times (RvT) was 4.9, 3.7 respectively. The RvT reached to 11.3 when treated with Cur (2.5 microg/ml) combined with EM (120 microg/ml). The DNR MFI in K562/A02 cells was significantly lower than that in K562 cells (P < 0.01), and was increased significantly when treated with Cur (2.5 microg/ml) or EM (120 microg/ml) (P < 0.05). There was no significant difference between DNR MFI of K562/A02 cells treated with Cur (2.5 microg/ml) or EM (120 microg/ml). Immunohistochemistry showed that P-gp expression was significantly higher in K562/A02 cells than in K562 cells (P < 0.01), and was reduced in K562/A02 cells treated with each (P < 0.01), though being still higher than that in K562 cells (P < 0.01). P-gp expression of K562/A02 cells treated with each drug for 5 days were lower than that for 3 days (P < 0.01), and lowered further when treated with Cur and EM together (P < 0.01). Mdr1 mRNA level in K562/A02 cells was higher than in K562 cells (P < 0.01), and was decreased when treated with each of the drugs (P < 0.01). The mdr1 mRNA level of K562/A02 cells treated with Cur (2.5 microg/ml) plus EM (120 microg/ml) was decreased most significantly than that treated with other group of drugs. After 5 day treatment the mdr1 mRNA level of K562/A02 cells with Cur (2.5 microg/ml) was lower than that with EM 120 microg/ml (P < 0.01).</p><p><b>CONCLUSIONS</b>Either Cur or EM can partly reverse the multidrug resistance of K562/A02 cells and decrease the expression and function of P-gp in a time-dependent way. MDR reversing effect of Cur combined with EM is stronger than that of Cur or EM alone.</p>
Subject(s)
Humans , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Genetics , Metabolism , Antineoplastic Agents , Pharmacology , Cell Proliferation , Cell Survival , Curcumin , Pharmacology , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Drug Synergism , Epirubicin , Pharmacology , Erythromycin , Pharmacology , Gene Expression Regulation, Neoplastic , Immunohistochemistry , K562 Cells , Leukemia, Erythroblastic, Acute , Genetics , Metabolism , Pathology , RNA, Messenger , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Time FactorsABSTRACT
<p><b>OBJECTIVE</b>To study the histochemical staining in the diagnosis of osteosarcoma.</p><p><b>METHODS</b>To compare the effectiveness of picrosirius red, improved Ponceau trichrome and Masson trichrome staining methods on bone formation tissues in conventional osteosarcoma, paraosteal osteosarcoma, periosteal osteosarcoma, extraskeletal osteosarcoma, inflammatory myofibroblastic tumour, malignant fibrohistiocytoma, chondrosarcoma, fibrosis with ossification and calcification.</p><p><b>RESULTS</b>With modified Ponceau trichrome staining, bone formation tissues showed a homogenous, orange-red interblended with blue in color. From osteoid to mature bone the color changed from orange-red, light blue to dark blue. Fibrotic tissue was stained blue in color with striated appearance. Cartilage was not stained. Picrosirius red method gave bone formation tissues homogenous staining. Along with bone maturation, from osteoid tissue to mineralized bones, the color showed changes from light red, yellow, orange-red, red to dark purple. The cartilage demonstrated homogenous light red in color. Fibrous tissue stained red interblended with yellow in color, striated in shape. With Masson trichrome staining osteoid displayed pale blue and mineralized bone showed dark blue in color. Fibrotic tissue showed a striated blue staining.</p><p><b>CONCLUSION</b>The modified Ponceau trichrome and Picrosirius red staining methods are better than Masson trichrome to demonstrate bone formation tissue in osteosarcoma. The former two methods could be also used in study on bone formation.</p>
Subject(s)
Humans , Bone Neoplasms , Pathology , Histocytochemistry , Osteosarcoma , Pathology , Staining and Labeling , MethodsABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression level of inhibitor of apoptosis protein survivin gene in human brain glioma and its role in malignant proliferation and antiapoptosis of brain glioma.</p><p><b>METHODS</b>Eighty-three cases of brain glioma specimen was chosen, protein expression of survivin and proliferating cell nuclear antigen (PCNA) was investigated by immunohistochemistry streptavidin-biotin complex (SABC) method, the immunoreactivity score (IRS) of survivin and the proliferative index (PI) were counted. Apoptotic cells were screened by TdT-mediated dUTP-biotin nick-end labeling (TUNEL) method, and the apoptotic index (AI) of brain glioma was calculated.</p><p><b>RESULTS</b>The survivin IRS, PI and AI of brain glioma were 3.8 +/- 3.9, (28.4 +/- 19.5)% and (1.0 +/- 0.8)% respectively, and all of them were elevated with the increase of pathological grade of brain glioma (P < 0.01 for all). PI in survivin positive group was significantly higher than that in survivin negative group (P < 0.01), and PI was positively correlated with survivin IRS (r = 0.740, P < 0.01). There was no significant difference between AI in survivin positive group and that in survivin negative group (P > 0.05), however, AI was negatively correlated with survivin IRS (r = -0.307, P < 0.01).</p><p><b>CONCLUSIONS</b>Survivin is overexpressed in brain glioma, and which may play important roles in malignant proliferation and antiapoptosis of brain glioma.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Apoptosis , Brain Neoplasms , Genetics , Metabolism , Pathology , Cell Proliferation , Glioma , Genetics , Metabolism , Pathology , Immunohistochemistry , In Situ Nick-End Labeling , Inhibitor of Apoptosis Proteins , Microtubule-Associated Proteins , Genetics , Neoplasm Proteins , Genetics , Proliferating Cell Nuclear AntigenABSTRACT
<p><b>OBJECTIVES</b>To obtain high therapeutic effect and low toxicity single-chain immunotoxin against hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Human mutant tumor necrosis factor-alpha (mTNFalpha) was linked with the 3' end of humanized single-chain Fv against HCC (hscFv25) in pGEX4T-1 vector. The anti-HCC immunotoxin was expressed in Escherichia coli and identified by western blot. The primary tumor regression trial in nude mice bearing HCC was evaluated the targeting therapeutic value of hscFv25-mTNFalpha. The tumor tissues were stained by immunohistochemical with TNFalpha antibody.</p><p><b>RESULTS</b>The expression of single-chain immunotoxin hscFv25-mTNFalpha was 12% of total bacteria proteins. The tumor regression trials of hscFv25-mTNFalpha showed 5/5 effective. It had 2/5 completely remission and 3/5 partly remission. The therapeutic result of hscFv25-mTNFalpha was better than that of mTNFalpha (F=8.70, 0.05). The HCC tissue treated by hscFv25-mTNFalpha expressed TNFalpha positive reaction. The positive granule mainly existed in HCC cytoplasm.</p><p><b>CONCLUSION</b>The single-chain immunotoxin hscFv25-mTNFalpha has high therapeutic effect and low toxicity. It has potentialities for clinical application.</p>