目的:观察和探讨细胞角质素CK18慢性胆囊炎患者的胆囊组织和血清中的表达及其意义。方法:35例经腹腔镜胆囊切除的慢性结石性胆囊炎患者(27例女性患者,8例男性患者,年龄在55.65±13.48岁),将患者分为两个组,A组为患慢性非活动性结石性胆囊炎者(n=10),B组为患慢性活动性胆囊炎者(n=25),在细胞凋亡早期胱门蛋白酶分裂的CK18用M30细胞凋亡酶联免疫吸附测定,总细胞角蛋白18(从凋亡及坏死细胞中分离)用M65酶联免疫吸附测定。然后计算M30/M65结果:胱门蛋白酶分裂的CK18,特别是总CK18在胆汁中的表达远高于血清。在B组中,胱门蛋白酶分裂的CK18和总CK18的表达在胆囊组织和血清中表达差异相当大。在胆囊粘膜上皮细胞胱门蛋白酶分裂的CK18染色呈强阳性。结论:CK18在胆囊上皮细胞中表达。胱门蛋白酶分裂的CK18和总CK18在胆囊组织中的表达远高于血清中的表达。胱门蛋白酶分裂的CK18和总CK18的表达水平在活动性胆囊炎和非活动性胆囊炎中的表达并无明显差异。
Objective To investigate the association of the expression of CD15 mRNA with the invasion and prognosis of hepatocellular carcinoma (HCC) and the expression of nm23H1 mRNA. Methods In situ hybridization and immunohistochemistry methods were used to detect the expression of CD15 mRNA and protein nm23H1 mRNA in HCC.Results In 99 cases of HCC, the positive rate of CD15 mRNA,its protein and nm23H1 mRNA were 38.4%, 36.4% and 76.8%, respectively. The expression of CD15 mRNA was consistent with its protein and negatively correlated with the expression of nm23H1 mRNA. The expression of CD15 mRNA and its protein, nm23H1 mRNA were associated with the invasiveness and metastasis of HCC and the prognosis of HCC patients. Conclusion The detection of CD15 expression could be a new pathological biology index to judge the metastasis and prognosis of HCC.
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 of costimulatory molecules, Fas/FasL, and major histocompatibility complex (MHC)-class II antigens in normal ocular tissues. Methods Twelve eyes were obtained from the eye bank of Zhongshan Ophthalmic Center within 16 to 24 hours postmortem. Six eyes were used for making the retinal wholemounts, and the tissues (iris and ciliary body, choroid, and retina) of the others were used for making the frozen sections. Immunohistochemical staining was carried out on these retinal wholemounts as well as on tissue sections to investigate the exprenion of B7-1 and B7-2 (costimulatory molecules), HLA-DR(MHC class II), CD68 (macrophages), Fas/FasL. Results The results of immunohistochemical staining showed the presence of B7-2, FasL, CD68 and HLA-DR in the iris and ciliary body. Expression of B7-1, FasL, CD 68, and HLA-DR was found in the choroid while HLA-DR, CD68 and FasL were detec table in the retina. Conclusion Expression of costimulatory molecu les, MHC-class II molecules and molecules related to apoptosis is different in the iris, ciliary body, choroid, and retina, which may play an important role in the stability of the immunological microenvironment of these tissues.(Chin J Ocul Fundus Dis，2003，19：109-112)
【Abstract】ObjectiveTo investigate the relationship between expression of Seprase and the clinicopathologic characteristics in colorectal cancer. MethodsThe expression of Seprase in 50 cases of colorectal cancer was detected with immunohistochemistry, Western-blotting technique and semi-quantitative immunohistochemistry, and the relationship between the expression of Seprase and the clinicopathologic characteristics (age, gender, tumor location, tumor size, gross appearance, depth of infiltration, histological classification, lymph node metastasis, venous infiltration, Dukes stage, distant metastasis) was analyzed. ResultsThe expression of Seprase was found both in tumor cells and adjacent stromal cells. The level of Seprase protein was higher in cancer tissue than that in normal tissue, and a semiquantitative assessment of the immunohistochemistry revealed a significant correlation between Seprase expression and the Dukes stage and the lymph node metastasis. ConclusionThe abundant expression of Seprase in colorectal cancer tissue is associated with the Dukes stage and the lymph node metastasis.
Objective To study the migration of Schwann cells from the nerve autograft in the acellular nerve allograft of the rats in vivo. Mehtods The sciatic nerves (20 mm long) of the SD rats were harvested and prepared for the acellular nerve grafts by the chemical extraction. Then, they were observed by the gross view, HE staining, and Antilamininstaining, respectively. Another 32 female SD rats weighing 250-300 g were obtained for the study. A 2-mm-long nerve autograft was interposed between the two 10-mm-long nerve allografts to form a 22-mm-long composite. Then, the composite was placed in the muscle space, together with a sole 22-mm-long nerve allograftas a control. They were harvested at 5，10，15 and 20 days, respectively, and were then given the HE staining and the S-100 staining. Results The acellular nerve graft was semitransparent under the gross view. HE staining showed that no cell was observed within the nerve graft. Anti-laminin staining showed that the basal membrane was partially interrupted, with a positive result (dark brown). All the nerve grafts in both the groups exhibited the existenceof the cells. The S-100 positive cells were observed from the 15th day at the far ends of the two allografts of the composite; however, there were no suchcells observed within the sole nerve allograft. Conclusion Schwann cells from the sciatic nerves (2 mm- long) of the rats can migrate in the acellular nerve allograft to the far ends of the neighboring 10-mm-long nerve allografts at 15 days after operation, which offers the theoretical basis forthe repair of the longrange nerve defect by the composite of the acellular nerve allografts with the interposed nerve autograft.
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 The major obstacle in pig to human transplantation is acute and hyperacute rejection (HAR) triggered mainly by alpha-galactosyl residues(alpha-Gal) in donor. Since the inbred-line Banna pig(IBNP) and Wuzhishan pig (IWZSP) are highly inbred and may be the potential donor for xenotransplantation, it is important to investigate the reaction between human serum and inbred-line pig tissues as well as the distribution of alpha-Gal in these tissues. METHODS Samples from heart, liver, spleen, lung, kidney, pancreas, small intestine, thymus, skin, lymph node and blood vessels at all levels were collected from four 8 to 11-month-old male IBNPs and one IWZSP. Affinity-immunohistochemistry assays were conducted following routine procedures on paraffin sections with normal human sera of blood type A, B, O, AB and BSI-B4(alpha-Gal specific binding lectin) as the primary antibodies or affinity reagents. Sections digested by alpha-galactosidase were also examined as control. RESULTS Parallel results were obtained from these pig tissues stained against human sera and BSI-B4. There was no significant difference both in the antigens recognized by sera of different blood types or BSI-B4 and in the distribution of alpha-Gal. The best alpha-Gal positive staining was appeared in vascular endothelial cells at all levels and partial parenchyma cells. However, tissues of cartilage, peripheral nerve and muscle were negative. After digested by alpha-Galactosidase, all samples were negative against BSI-B4 and human sera except few positions that showed different staining. CONCLUSION The distribution of target antigen is similar in various tissues of the two kinds of pigs. Though alpha-Gal is the major xenoantigen in IBNP and IWZSP, there may be some unknown antigens related to pig to human transplantation. Possibly the level and distribution of antigen expression in pig tissues are not the first affair to be considered, and these pigs should be genetically modified in order to eliminate rejection in pig to human xenotransplantation.
ObjectiveTo explore the expressions of glucose regulated protein (GRP) 78 and GRP94 in rectal adenocarcinoma and their clinical significances. MethodsIn 45 paraffin-embedded sections of rectal adenocarcinoma tissues and adjacent normal tissues, the expressions of GRP78 and GRP94 were examined by EnVisionTM. ResultsThe positive rates of GRP78 and GRP94 protein expression in the rectal adenocarcinoma tissues were 53.33% (24/45) and 53.33% (24/45), while those in the adjacent normal tissues were 13.33% (6/45) and 15.56% (7/45), respectively. There was a statistical significance of the expression of GRP78 or GRP94 between the tumor tissues and the adjacent normal tissues (all P < 0.001), and it was found that the positive expression rates were relevant to the extent of differentiation, Dukes stage, and lymph node metastasis of cancer (all P < 0.05), but not to the patient's sex, age, and size of tumor (all P > 0.05). And there was a statistical significance in Spearman method about the rate of positive expression between GRP78 and GRP94 (rs=0.464, P < 0.01). ConclusionGRP78 and GRP94 protein is highly expressed in rectal adenocarcinoma tissue, related to its metastasis and invasion, might be used as a useful indicator to judge the malignant degree.