OBJECTIVE: To observe the protein expression of phosphorylated form of P38 mitogen-activated protein kinase(P38MAPK) and c-Jun in hypertrophic scar skin and to explore their influences on the formation and maturation of hypertrophic scar. METHODS: The expression intensity and distribution of phosphorylated form of P38MAPK and c-Jun were examined with immunohistochemistry and pathological methods in 16 cases of hypertrophic scar skin and 8 cases of normal skin. RESULTS: In normal skin, the positive signals of phosphorylated form of P38MAPK mostly distributed in basal lamina cells of epidermis, while c-Jun was mainly located in epidermal cells and endothelial cells. The positive cellular rates of two proteins were 21.3% +/- 3.6% and 33.4% +/- 3.5% respectively. In proliferative hypertrophic scar skin, the particles of phosphorylated P38MAPK and c-Jun were mainly located in epidermal cells and some fibroblasts. The positive cellular rates of two proteins were significantly elevated to 69.5% +/- 3.3% and 59.6% +/- 4.3% respectively (P lt; 0.01). In mature hypertrophic scar, the expression of these proteins decreased but was still higher than that of normal skin. CONCLUSION: The formation and maturation of hypertrophic scar might be associated with the alteration of phosphorylated P38MAPK and c-Jun protein expression in hypertrophic scar.
Objective To investigate the expression of enhancer of zeste homolog 2 （EZH2） mRNA in colorectal cancer tissues， and to explore the relationship with clinicopathological features of it. Methods PCR was used to examine the expression of EZH2 mRNA in colorectal cancer tissues， paracancerous tissues， and normal epithelium tissues of 27 patients with colorectal cancer. Meanwhile， the relationship between the expression of EZH2 mRNA and clinicopathological features of colorectal cancer were analyzed. Results Expression level of EZH2 mRNA in colorectal cancer tissues， paracancerous tissues， and normal epithelium tissues were 3.01±1.29， 1.20±0.69， and 0.87±0.37 respectively， and the expression value of colorectal cancer tissues at the top （P＜0.01）， but there were no significant difference between the paracancerous tissues and normal epithelium tissues （P=0.403）. Expression of EZH2 mRNA were related to TNM stages （P=0.002） and degree of differentiation （P=0.005）， but were not related to age （P=0.388）， gender （P=0.963）， diameter of tumor （P=0.579）， histological type （P=0.945）， location of tumor （P=0.611）， and lymph node metastasis （P=0.449）. Conclusions EZH2 mRNA expresses highly in colorectal cancer tissues， so it may play an essential role in the emergence and development of colorectal cancer. EZH2 mRNA can be used as one of the referenced indexes to determine the malignant degree and process of colorectal cancer.
In order to study the expression change of insulin-like growth factor-1 (IGF-1) and its receptor genes in different generations of tendon cell in culture, Dig-labeled synthesized oligonucleotide probes were used to detect the mRNA expression in primary, 6th and 13th generation of tendon cell. The results showed that IGF-1 receptor mRNA was expressed in all of the 3 above generation tendon cells. IGF-1 mRNA was expressed only in primary and 6th generation cells. Tendon cell of 13th generation did not express IGF-1 mRNA. It might suggest that the absence of IGF-1 mRNA expression be one of the causes which led to the decrease of reproductive ability of 13th generation tendon cell.
Objective To investigate the expression levels of osteoprotegerin (OPG) and receptor activator of NF-κB l igand (RANKL) mRNAs in bone tissues of the femoral head of the patients suffering glucocorticoid-induced osteonecrosisof the femoral head (ONFH), and to discuss the relationship between OPG/RANKL and ONFH. Methods Between March2007 and March 2008, bone tissues of the femoral head were collected as the experimental material from 35 patients suffering ONFH (experimental group) and from 21 patients suffering fracture of femoral neck (control group). The ratio of men to women in both groups was 4 ∶ 3, whose age was 41-70 years old (55.34 on average in the experimental group and 55.33 on average in the control group). The experimental group received over 3 weeks’ glucocorticoid treatment or more than 1 week’ s high-dose glucocorticoid treatment in recent 2 years, while the control group never received more than 1 week’s hormone treatment. In the two groups, the microstructure of bone tissues of the femoral head was detected by HE staining and the bone tissue total RNA was extracted, and then the expression levels of OPG mRNA and RANKL mRNA were examined by realtime quantitative PCR (RTQ-PCR) for each sample. Results HE staining: bone trabeculae and bone units were replaced by interrupted bone fragments, which were surrounded by many inflammatory granulation tissues and few osteocytes were seen in bone lacunae in the experimental group. In the control group, bone trabeculae and bone units were made by complete lamellar bones which surrounded blood vessels and osteocytes were seen in lacunae. RTQ-PCR testing: in the experimental group, OPG mRNA and RANKL mRNA were 1.35 ± 0.42 and 4.36 ± 1.35, respectively, while in the control group they were 1.78 ± 0.63 and 3.49 ± 1.02, respectively. The expression level of OPG mRNA in the experimental group was significantly lower than that in the control group, and the expression level of RANKL mRNA of the former was significantly higher than the latter. The OPG mRNA/ RANKL mRNA ratio in the xperiment group (0.34 ± 0.16) was significantly lower than that in the control group (0.54 ± 0.20), and there was significant difference (P lt; 0.05). Conclusion The glucocorticoid-induced ONFH may be related to the expression levels of OPG mRNA/RANKL mRNA in bone tissues.
Objective To construct the recombined DNA pcDNA3.1-hBMP-2 and transfect into human marrow stromal stem cells (MSCs) in vitro, and to explore theeffects of transfection on cellular proliferation and expression of vascular endothelial growth factor (VEGF). Methods The expression of human bone morphogenetic protein 2(hBMP-2) in these cells after transfection was determined by in situ hybridization and immunohistochemical analysis and Western blot analysis. The changes of cell proliferation were observed by flow cytometry. The effects of BMP-2 gene transfection on expression of VEGF in the cells were analyzed by in situ hybridization of VEGF cDNA probe. Results Stable expressionof hBMP-2 in pcDNA3.1-hBMP-2 transfected MSCs was confirmed in the levels of mRNA and protein.Cellular proportion in S period increased, which indicated that the synthesis of cell DNA increased. The expression of VEGF in the cells increased obviously. Conclusion With the help of lipofectamine, the pcDNA3.1-hBMP-2 were transfected into human MSCs successfully. hBMP-2 plays an important role in promoting cellular proliferation and vascular generation during bone repair.
Objective To observe the expression and localization of cellular homolog FLICE-like inhibitory protein (c-FLIP) in the procedure of benign biliary stricture formation and discuss the significances. Methods The method of in situ hybridization was used in anastomotic tissues from 15 dogs (experimental group) in 2, 3, 4, 5, 6 months after bile duct injury and 15 matching sham operation dogs (sham operation group) for analyzing the expression and localization of c-FLIP and calculating the average integrated optical density of each slice. Stain cells were counted under the magnification field (×400) and at least 5 fields per slice were examined. The cells stained red in the nuclei and (or) the cytoplasm were positive cells. The signals meant: Negative for cells no stained, weak positive for the cells with nuclei and (or) cytoplasm stained pink; b positive for the cells stained the bright red; while middle positive for the cells stained between the both. The image analysis software （Image pro plus 4.5） was applied in the gland tissue and interstitial tissue in each slice to calculate the average integrated optical density for the expression of c-FLIP. Results In the experimental group, there were all b positive expressions of c-FLIP in the interstitial tissue at all the time points, mainly expressed in the cytoplasm of fibroblast and very little or almost no expression in the glandular tissue. Positive expression of c-FLIP in the interstitial tissue was significantly ber than that of gland tissue (Plt;0.05); There were no significant differences among each time point in either the interstitial tissue or gland tissue (Pgt;0.05). In the sham operation group, there were all weak positive expressions of c-FLIP in the interstitial tissue and gland tissue at all the time points and was no significant difference (Pgt;0.05), no difference between each phase (Pgt;0.05). The expression of c-FLIP in the experimental group was significantly higher than that in the sham operation group in the interstitial tissue at all the time (Plt;0.05), while no significant that in the gland tissue (Pgt;0.05). Conclusion After bile duct injury, the expression of c-FLIP in anastomotic interstitial tissue is sustainable, by which the continuing obstruction effect to apoptosis may have a close relationship with the formation of biliary benign stricture.
Objective To observe the effects of vascular endothelial growth factor antisense oligonucleotide (VEGF-ASODN) on expression of vascular endothelial growth factor (VEGF) and growth in gastric cancer cells. Methods The VEGF-ASODN was synthesized artificially with phosphorothioic acid. After transfecting with VEGF-ASODN in gastric cancer cells SGC-7901, the initial copy number of mRNA was detected by real-time RT-PCR, and the quantity of VEGF protein in both cell and supernatant were detected by ELISA. The levels of expression of survivin protein in cells were measured by Western blot. FCM and MTT method were used to detect cellular apoptosis and the activity of cells, respectively. The effect of transfection on the growth of cells was evaluated by growth curve. Results The copy number of VEGR mRNA, protein levels of VEGF in the cells and in culture fluid all decreased when the concentration of transfected VEGF-ASODN increased, as well as the levels of survivin protein （P＜0.05）. The ratio of apoptosis increased, the activity of cells also decreased as the concentration of transfected VEGF-ASODN increased （P＜0.05）. Conclusion Transfection with VEGF-ASODN in gastric cancer cells SGC-7901 can inhibit the expressions of VEGF and survivin remarkably. It can enhance cellular apoptosis and suppress growth of cells.
ObjectiveTo detect the expression of motilin in gastric cancer tissues and to explore the relationship between motilin protein expression and clinicopathologic characteristics of gastric cancer. MethodsThe immunohistochemical staining was used to detect the expression of motilin protein in gastric cancer, paracancerous tissues, and normal gastric mucosa tissues. The relationship between motilin protein expression and clinicopathologic characteristics of gastric cancer was analyzed. ResultsThe expression of motilin protein in gastric cancer tissues (1 206.43±631.67) was significantly higher than that in normal gastric mucosa tissues and paracancerous tissues, respectively (Plt;0.01). The difference of motilin protein expression between normal gastric mucosa tissues and paracancerous tissues was not significant (Pgt;0.05). The expression of motilin protein in gastric cancer was correlated with the site of tumor, differentiation degree, and lymph node metastasis (Plt;0.05). ConclusionMotilin may participate in the carcinogenesis of gastric cancer, and correlated with the invasion and metastasis of gastric cancer.
Objective To screen the differentially expressed genes and pathways involved in rosacea using bioinformatics analysis. Methods The GSE65914 gene chipset was collected from the Gene Expression Omnibus (up to July 12th, 2021). It was searched according to the keyword “rosacea”. The data was analyzed by GEO2R platform. The common differential genes of three subtypes of rosacea were screened out. The online DAVID analysis tool was used to perform the gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Protein-protein interaction networks of differentially expressed genes were made by String and Cytoscape. The key modules and genes were screened by Mcode and Cytohubba. Results A total of 957 common differential genes were identified, including 533 up-regulated genes and 424 down-regulated genes. GO enrichment analysis showed that these genes were mainly involved in immune response, inflammatory response, intercellular signal transduction, positive regulation of T cell proliferation, chemokine signaling pathways, cell surface receptor signaling pathways, cellular response to interferon-γ, and other biological processes. KEGG pathway enrichment analysis mainly included cytokine-cytokine receptor interaction, rheumatoid arthritis, chemokine signaling pathway, PPAR signaling pathway, Toll-like receptor signaling pathway, nuclear transcription factor-κB signaling pathway, tumor necrosis factor signaling pathway and other signaling pathways. Cytohubba analysis revealed 10 key genes, including PTPRC, MMP9, CCR5, IL1B, TLR2, STAT1, CXCR4, CXCL10, CCL5 and VCAM1. Conclusion The key genes and related pathways may play an important role in the pathogenesis of rosacea.
The expression and rearrangement of bcl-2 gene in 64 cases with colorectal carcinoma were studied by immunohistochemical technique and semi-nest PCR respectively. The results showed the abnormal changes of the expression and rearrangement of bcl-2 gene had emerged in the early stage of colorectal carcinoma. The tumors with the expression of bcl-2 were associated with a higher incidence of metastasis to lymphatic node. The rearragement of bcl-2 was significantly higher in late-stage than that in early-stage. These suggest that bcl-2 gene involves in the regulation of the development of colorectal carcinoma. The state of the changes of bcl-2 gene in colorectal carcinoma may predict the therapeutic effect and prognosis of colorectal carcinoma.