ObjectiveTo observe the expression of connective tissue growth factor (CTGF) in injured model of retinal pigment epithelial (RPE) cells and the promoting effect of CTGF on migration of RPE cells.MethodsCultured monolayer-confluent human RPE cells were scraped with a trephine and a cotton stick, and set up the injured model of RPE cells with round scraped area. Immunohistochemistry and in situ hybridization(ISH) were used to detect the expression of CTGF protein and mRNA in injured RPE cells at distinct time points after injury. The number of RPE cells migrated to injured area was measured and the effect of CTGF on migration of RPE cells and the effect of dexamethasone (DEX) on the promoting process of CTGF were observed.ResultsThe results of immunohstochemistry and ISH indicated the weak positive expression of CTGF in RPE cells at the edge of scrape 6 hours after injury, and the positive expression increased gradually as time goes by after the injury. Strong positive expression of CTGF in RPE cells at the edge of scrape was found 24 and 48 hours after injury. Rebuilt human CTGF stimulated migration of RPE cells in a dose-depended manner, and DEX significantly inhabited the migration.ConclusionCTGF involves in the procedure of repair of injury of RPE cells, which may play an important role in the pathogenesis of intraocular proliferative diseases such as proliferative vitreoretinaopathy.(Chin J Ocul Fundus Dis, 2005,21:306-309)
Objective To study the association and the effect of the expression of p16 and p53 protein on the occurence and development of gallbladder carcinoma. Methods The expression of p16 and p53 protein were detected in 40 cases of gallbladder carcinoma with immunohistochemical method. Results The expressions of p16 and p53 protein were closely correlated to the tumor pathological grade, lymph mode metastasis and prognosis. p16 protein was correlated to the Nevin classifications. Conclusion The results indicate that the low expression rate of p16 protein occurred in the advanced stage of gallbladder carcinoma. The expression of p16 and p53 protein are helpful in judging the malignant degree and prognosis of primary gallbladder cancer.
Objective To investigate the expression and clinical significance of S100A4 protein in tumorstroma of nonsmall cell lung cancer（NSCLC） to study its correlation with invasion, metastasis and prognosis. Methods Immunohistochemical staining（SP method）for S100A4 protein expression was performed in tissue sections from 130 patients with NSCLC operated and to analyze association of S100A4 protein with clinicopathological parameters in lung cancer and prognosis. Results The total positive expression rates of S100A4 protein in stroma of NSCLC was 72.3%. The positive expression rates of S100A4 protein in stroma of squamous cell carcinoma, adenocarcinoma, adenosquamous carcinoma and large cell lung cancer were 84.3%，59.6%，70.0% and 75% respectively.The expression of S100A4 protein was significantly associated with lymph node metastasis (χ2=18.91, P=0.000), distant metastasis(χ2=5.51, P=0.019) and TNM stage (χ2=21.54, P=0.000). The 3 years survival rates of patients whose tumourstroma stained positive for S100A4 was lower than that of patients whose tumourstroma stained negative (36.2% vs. 63.9%, P=0.003). Cox’ risk ratio model analysis indicated that age ≤50 years (OR=1.866), lymph node metastasis(OR=1.826), distant metastasis(OR=6.224), lower histology differentiation and undifferentiation (OR=1.793), TNM stage Ⅲ-Ⅳ (OR=2.573) and positive expression of S100A4 protein in stroma of NSCLC(OR=1.776) were significantly independent prognostic factors which affected survival. Conclusion Expression of S100A4 protein in stroma of NSCLC is significantly associated with invasion, metastasis, TNM stage and prognosis. S100A4 protein might become a marker for prediction of tumor progression of disease and clinical therapy.
Objective To study the relationships between expressions of somatostatin receptor subtypes（SSTR1-SSTR5） and angiogenesis in colorectal cancer. Methods The expressions of SSTR1-SSTR5, VEGF, and CD34 in the paraffin sections of colorectal cancer tissues from 127 cases were detected by the standard streptavidin-peroxidase (SP) technique. CD34 was used as a marker to account microvessel density (MVD) in colorectal cancer tissues. The relationships between the expressions of SSTR1-SSTR5 and VEGF expression, or MVD were analyzed. Results The positive expression rate of SSTR1, SSTR2, SSTR3, SSTR4, and SSTR5 was 64.6% （82/127）, 36.2% （46/127）, 18.9% （24/127）, 18.9% （24/127）, and 38.6% （49/127） in colorectal cancer tissues, meanwhile, the positive expression rate of VEGF was 63.8% （81/127） and MVD was (34.67±16.62)/HP in colorectal cancer tissues. The positive expression rate of VEGF (47.8%， 22/46) and MVD 〔(29.00±15.32)/HP〕 in colorectal cancer tissues with SSTR2 positive expression were significantly lower than those in colorectal cancer tissues with SSTR2 negative expression 〔72.8%， 59/81； (37.90±16.56)/HP〕, Plt;0.05. There were no relationships between SSTR1, SSTR3, SSTR4, and SSTR5 expression and VEGF expression or MVD （Pgt;0.05）. Conclusion The positive expression of SSTR2 is related with angiogenesis in colorectal cancer tissues.
OBJECTIVE: To investigate the changes of fibroblast growth factor (FGF) in burn wounds. METHODS: The FGF expression in the center of wound granulation, the edge of wound, the healed part of wound, the normal skin of patients, and the heal course of second degree burn wounds were detected by immunohistochemical methods. RESULTS: The expression intensity of FGF was different in the different sites of third degree burn wounds. The highest contents of FGF was in the center granulation of burn wounds, the less was in the borderline of wound and healed skin, and the least was in the healed skin. FGF expression mainly concentrated in the middle layer of wound, and almost no FGF expression in normal skin. The most FGF expression was occurred at 14 days after injury in second degree of burn wound. CONCLUSION: The changes of FGF in wounds are closely related to the wound healing, and rational use of FGF can promote wound healing.
Objective To observe the expression and distribution of transforming growth factor-β1 (TGF-β1) in the healing process of bile duct and discuss its function and significance in the process of benign biliary stricture formation. Methods An injury to bile duct of dog was made and then repaired. The expression and distribution of TGF-β1 in the tissue at different time of the healing process were studied after operation with immunohistochemical SP staining. Results TGF-β1 staining was observed in the granulation tissue, fibroblasts and endothelial cells of blood vessels. High expression of TGF-β1 was observed in the healing process lasting for a long time. Conclusion The high expression of TGF-β1 is related closely with the fibroblast proliferating activity, extracellular matrix overdeposition and scar proliferation in the healing process of bile duct.
To investigate the relationship between metallothionein (MT) and prognosis in breast cancer MT expression was determined with immunohistochemical method (SABC). Results: There was a statistically significant association between expression of MT in breast benign and malignant disease (Ｐ＜0.005). The positive rate was 73.8%(62/84) and 15.0%(3/20) in breast cancer and mastofibroma respectively. The positivity of MT was ber in advanced clinical stages than in early clinical stages. There was no association between MT expression and lymph node metastasis. The mortality of the cancer cases with lymph node metastasis having positive MT expression was higher than those with negative MT expression. Conclusion: MT can be taken as a prognostic index of breast cancer.
Objective It is reported that transforming growth factor β1 (TGF-β1) has the protective effects on the articular cartilage in osteoarthritis (OA). To investigate the significance of the expressions of matrix metalloproteinase 9 (MMP-9), TGF-β1 mRNA and corresponding proteins in OA. Methods The specimens of articular cartilage and synovium were collected from voluntary donators, including 60 cases of OA (experimental group) and 20 cases of traumatic amputation,cruciate l igament rupture, discoid cartilage injury, and menisci injury (normal control group). The pathological changes were observed by HE staining. MMP-9 and TGF-β1 protein expressions were detected by immunohistochemical technique, and the mRNA expressions of MMP-9 and TGF-β1 were detected through in situ hybridization technique; and their correlation was analysed. Results HE staining showed: shrinkage, necrosis, and irregular arrange of the articular chondrocytes, extracellular matrix fracture, hypertrophy and hyperplasia synovium, infiltration of lymphoid and mononuclear cells and prol iferation of many small blood vessels in the experimental group; regular arrangement of the articular chondrocytes, the homogeneously staining matrix, and synovial tissue without chronic inflammation and significant prol iferation in the normal control group. The mRNA and protein expressions of MMP-9 and TGF-β1 were positive in 2 groups. The positive-stained cells included chondrocytes, synovial l ining cells, and vascular endothel ial cells, fibroblasts, and inflammatory infiltrated cells in subsynovial layer. The expressions of mRNA and corresponding protein of MMP-9 and TGF-β1 in the experimental group were significantly higher than those in the normal control group (P lt; 0.01). There was a positive correlation between MMP-9 mRNA and protein expression (r=0.924, P=0.000), and between TGF-β1 mRNA and protein expression (r=0.941, P=0.000) in the experimental group. There was a negative correlation between the expression of MMP-9 protein and TGF-β1 protein (r= — 0.762, P=0.000), and between the expression of MMP-9 mRNA and TGF-β1 mRNA (r= — 0.681, P=0.000) in the experimental group. Conclusion The higher expression of TGF-β1 can protect articular cartilage by down-regulating the expression of MMP-9 of chondrocytes and synoviocytes in OA, which may delay the biological behavior of OA such as occurrence and progress, etc.
Objective To investigate the possible interaction between the ras and p53 genes overexpression in thyroid carcinoma, and whether there is correlation between the ras and p53 overexpression and clinico-pathological criteria. Methods Thyroid lesions from eighty patients were examined for expression of ras and p53 genes by the LSAB immunohistochemistic method. Of these patients, 54 were diagnosed as malignant lesions and 26 benign nodular thyroid disorders. Results The positive immunostain rate for ras and p53 genes was 90.7%, 23.0% and 55.5%, 30.7% in carcinoma and benign lesions respectively with statistically significance between thyroid carcinomas and benign disorders (Ｐ＜0.05). Both ras and p53 overexpressions coexisted in 30 thyroid carcinomas and follow-up showed that 3 of them died and 5 of them had recurrence within 4 years.Conclusion Activation of ras gene and inactivation of p53 gene are cooperatively associated in thyroid tumorigenesis. The concurrent overexpression of ras and p53 could result in a poor prognosis.
ObjectivesTo investigate the expression of E-selectin of adhesive molecule in microvessels in rats′ retina with diabetic retinopathy (DR) at the early, middle, and late phase.MethodsNinety Sprague Dawley (SD) male rats were randomly divided into normal control group (6, 9, and 12 months subgroups with 10 rats in each subgroup) and streptozotocin (STZ) DR group (6, 9, 12 months subgroups with 20 rats in each subgroup). The DR model was set up by one-off celiac injection with STZ at a dose of 60 mg/kg. The rats were executed according to the different time points, and the eyeballs were removed and the digested stretched preparations of retinal microvessels were made. E-selectin of adhesive molecule in microvessels was revealed by EnVision immunohistochemical staining, and the expression of E-selectin in retinal microvessels and the morphological changes were analyzed and observed by Leica-Q550W computer image analytical equipment. The relativity between degree of impairment of DR and expression of E-selectin was detected.ResultsMorphological changes: hyperplasia of endotheliocytes and decreased pericytes were found in 6month DR group; irregularly arranged retinal capillary networks, obviously thickened basilar membrane, many aggregation and adhesion of leucocytes to capillary walls, large areas of acellular capillaries, and opoptosis of endotheliocytes and pericytes were found in 9-and 12-month DR group. Results of quantitative analysis: there was no expression of E-selectin in normal control group, while expression of E-selectin with different degree in each DR group was found. There was significance difference among DR subgroups (Plt;0.01), and the highest expression was found in 6-and 12-month DR group.ConclusionsThe expression of adhesive molecule in microvessels in rats' retina increases as the disease course of DR extends, and has positive correlation with the degree of impairment of microvessels in DR. (Chin J Ocul Fundus Dis, 2005,21:318-321)