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 Methods Ninety male Wister rats were randomly divided into normal control group, diabetic group and FTY720 group, thirty rats in each group. Diabetes was induced by giving a single intraperitoneal injection of streptozocin. FTY720 group was administered with FTY720 at a dose of 0.3 mg/kg by oral gavage daily for 3 months after establishment of diabetes. All rats were used for experiments following intervention for 3 months in FTY720 group. Immunohistochemical staining was used to observe the expression and distribution of intercellular adhesion molecule (ICAM-1) and vascular cell adhesion molecule (VCAM-1), and the positive cells were counted. Real-time reverse transcription PCR was used to measure mRNA expression of ICAM-1 and VCAM-1. Fluorescein isothiocyanate-Concanavalin A perfusion was used to detect retinal leukocytes adhesion. Evans blue (EB) perfusion was used to analyze retinal vascular permeability. Immunofluorescence staining was used to detect retinal inflammatory cells infiltration. Results In diabetic group, both ICAM-1(t=12.81) and VCAM-1 (t=11.75) positive cells as well as their mRNA expression (t=16.14, 9.59) were increased compared with normal control group, with statistical significance (P < 0.05). In FTY720 group, both ICAM-1(t=-9.93) and VCAM-1 (t=-6.61) positive cells as well as their mRNA expression (t=-15.28, -6.10) were decreased compared with diabetic group, with statistical significance (P < 0.05). Retinal leukocytes adhesion (t=16.32) and EB permeability (t=17.83) were increased in diabetic group compared with normal control group, while they were decreased in FTY720 group compared with diabetic group(t=-9.93, -11.82),with statistical significance (P < 0.05). There were many CD45 positive leukocytes infiltration in retina of diabetic group, including CD11b positive macrophage/activated microglia, while both of them were little in FTY720 group. Conclusions FTY720 can decrease retinal leukocytes adhesion, reduce retinal vascular permeability and inflammatory cells infiltration, which is associated with down-regulation of ICAM-1 and VCAM-1.
【Abstract】ObjectiveTo study the expressions of matrix metalloproteinase 2 (MMP2) and carbohydrate antigen 50 (CA50) in colorectal carcinoma, cancer-adjacent mucosa （2 cm from the nether edge of tumor）, cancerdistant mucosa （5 cm from the nether edge of tumor） and normal colorectal mucosa, and to elucidate their effects on the development of colorectal carcinoma. MethodsThe expressions of MMP2 and CA50 were detected immunohistochemically in 40 cases of colorectal carcinoma, cancer-adjacent mucosa, cancer-distant mucosa and 10 cases of normal colorectal mucosa. Results①The expression intensity and positive rates of MMP-2 and CA50 increased significantly in turn by normal mucosa, cancer-distant mucosa, cancer-adjacent mucosa and colorectal carcinoma. ②The expression of MMP2 was correlated with CA50 in colorectal carcinoma. ③The expression of CA50 in colorectal carcinoma was closely associated with tumor differentiation, and the expression of MMP2 in colorectal carcinoma was closely associated with differentiation and Dukes stages as well. ConclusionOver expression of MMP2 facilitates the malignant progress of colorectal carcinoma; CA50 is a reliable marker of malignance in colorectal carcinoma; CA50 and MMP2 may have synergetic effects on the development of colorectal carcinoma.
To study the mechanism of p16,Cyclin D1 and CDK4 and their relationship with pancreatic carcinoma, their expressions were examined by immunchistochemistry methods. Results: overpression of Cyclin D1 and CDK4 was revealed in these samples and p16 was undertectable. There was a negitive correlation between p16 and Cyclin D1 (Ｐ＜0.05), and a positive relation between Cyclin D1 and CDK4 (Ｐ＜0.05). The results indicate that abnormality of p16, Cyclin D1 and CDK4 may be involved in the molecular mechanism of pancreatic carcinoma, p16 lower expression and Cyclin D1 over expression may coexit in the development of pancreatic carcinoma.
Objective To observe the changes of immune status in recipient after implanting with xenogeneic acellular bone matrix (ACBM). Methods Twenty rabbits were randomly divided into 4 groups. Autograft，ACMB and bone soaked in alcohol were implant into the 3 experimental groups separately, and No-treatment was done as control group. The CD4+,CD8+,CD25+T lymphocytes in blood were detected by flow cytometer at 1, 2, 4 and 6 weeks after operation. After 2 and 6 weeks of implantation, the changes of bone and tissue were observed by histology. Results After 2-6 weeks, CD4+ and CD8+ T cells were significantly higher in the implantedgroup of bone soaked in alcohol than that in the other 3 groups(Plt;0.05) and there wasno statistically significant difference in the other 3 groups(Pgt;0.05). After 2 weeks, CD25+ T cells were significantly higher in the implanted group of bone soakedin alcohol than that in the other groups. In the 2nd week, there were inflammatory infiltration with a predominance of granulocytes. In the 6th week, there were many fibroblasts instead of granulocytes with a few lymphocytes and cartilage island formed in the implanted groups of autograft and ACMB. Conclusion ACBM implanting has low influence on cellular immunity in recipient.
ObjectiveTo evaluate the effect of pre-infusion of allogeneic lymphoyctes treated with 5-FU on the rat liver graft. MethodsRat liver transplant models from Wistar to SD were established. Four groups were designed as following: control group: only liver transplantation without any other intervention; lymphocytes group: 1 ml of untreated lymphocytes (5×106/ml) from Wistar rats were preinfused into SD rats on day 7 and 4 separately before transplantation; lymphocytes with low concentration of 5-FU group: low concentration 5-FU (7.5 μg) treated lymphocytes were preinfused as above; lymphocytes with high concentration of 5-FU group: high concentration 5-FU (15 μg) treated lymphocytes were preinfused as above. Fas-L and CD8 expression were detected by immunohistochemistry method on day 7 after transplantation. ResultsThe integral opticaldensity (IOD) of Fas-L positive lymphocytes in the lobules of liver and portal areas were higher in lymphocytes with low concentration of 5-FU group than in the other groups （Plt;0.05）. There was no difference between lymphocyte group and lymphocytes with high concentration of 5-FU group （Pgt;0.05）. The IOD of CD8+ expression in lobules of liver was not different among all the three lymphocytes treated groups （Pgt;0.05）. But in portal areas, CD8+ expression was lower in the lymphocytes with low concentration of 5-FU group than in the other groups （Plt;0.05）. ConclusionPreinfusion of lymphocytes treated with low concentration 5-FU can induce graft immune tolerance, the probable mecanism of which is the increasing Fas-L expression in graft.
Objective To investigate the histological origin, diagnosis, differential diagnosis and treatment of thyroid carcinoma showing thymus-like differentiation (CASTLE). Methods Five patients with thyroid CASTLE were adopted by surgical resection and postoperative radiotherapy, and the CD5, CD117, CK5/6, P63, thyroid transcription factor-1 (TTF-1), carcino-embryonic antigen (CEA), calcitonin (CT), Ki-67, chromogranin A (CgA), thyrobolulin (Tg), peroxisome proliferator activated receptorγ (PPAR-γ), sodium iodide symporter (NIS), and thyroid stimulating hormone receptor (TSHR) were detected in tumor tissues by immunohistochemistry S-P method and v-raf murine sarcoma viral oncogene homolog B1 (BRAF)V600E gene and telomerase reverse transcriptase (TERT) promoter mutations were detected by DNA sequencing. Eight cases of poorly differentiated thyroid carcinoma and 6 cases of anaplastic thyroid carcinoma were adopted by comprehensive comparative analysis. Results Thyroid CASTLE tumor cells showed the positive expression of CD5, CD117, CK5/6 and P63, and the negative expression of TTF-1, CT, CgA, Tg, PPAR-γ, NIS and TSHR. There were partly positive expression for CK5/6, P63, TTF-1, CgA, Tg, NIS and TSHR, and negative expression for CD5 and CD117 in the poorly differentiated thyroid carcinoma and anaplastic thyroid carcinoma. The BRAFV600E gene and TERT promoter mutations were not detected in thyroid CASTLE, and the BRAFV600E gene mutations were also not detected in the poorly differentiated thyroid carcinoma and anaplastic thyroid carcinoma. Four cases of poorly differentiated thyroid carcinoma showed the TERT promoter mutations (4/8) included 3 cases with C228T and 1 case with C250T. Two cases of anaplastic thyroid carcinoma showed the TERT promoter mutations (2/6) included 1 case with C228T and 1 case with C250T. There was no recurrence and metastasis after 3–47 months (an average of 25.6 months) of followed-up in thyroid CASTLE patients. Conclusions The histological origin of thyroid CASTLE may be not related to the thyroid. There is important clinical value to combined detection of CD5, CD117, P63, TTF-1, Tg, NIS, and TSHR for the diagnosis and differential diagnosis of thyroid CASTLE. The further study still need for the diagnosis and differential diagnosis of thyroid CASTLE according to the detection of BRAFV600E and TERT promoter mutations.
Objective To investigate the expressions and clinical significance of Notch-2 protein and Numb protein in papillary thyroid carcinoma (PTC). Methods PTC tissues and its para-cancerous tissues of 50 patients with PTC who treated in The Affiliated Hospital of Inner Mongolia University for Nationalities from Mar. 2014 to Mar. 2017 were retrospectively collectied, to detect the expressions of Notch-2 protein and Numb protein by immunohistochenmical method. Results ① Expressions of Notch-2 protein and Numb protein in PTC tissues and para-cancerous tissues: the positive-expression rate of Notch-2 protein in PTC tissues was 82.00% (41/50), which was higher than that of para-cancerous tissues〔18.00% (9/50)〕, the difference was statistically significant (χ2=40.960, P<0.001). The positive-expression rate of Numb protein in PTC tissues was 66.00% (33/50), which was higher than that of para-cancerous tissues 〔0 (0/50) 〕, the difference was statistically significant (χ2=49.254, P<0.001). ② The relationship between expression of Notch-2 protein and expression of Numb protein in PTC tissues: there was a positive correlation between expressions of Notch-2 protein and Numb protein in PTC tissues (rs=0.323, P=0.022). ③ The relationship between expressions of Notch-2 protein and Numb protein in PTC tissues and clinicopathological features of the PTC patients: the expression of Notch-2 protein in PTC tissues was not significantly correlated with gender, age, tumor diameter, capsule infiltration, cervical lymph node metastasis, and TNM staging (P>0.05). The expression of Numb protein in PTC tissues was not significantly correlated with gender, age, tumor diameter, and capsule infiltration (P>0.05), but was significantly correlated with cervical lymph node metastasis and TNM staging (P<0.05), the positive rates of Numb protein in patients of staging Ⅲ+Ⅳ group and cervical lymph node metastasis group were lower than those of patients in staging Ⅰ+Ⅱ group and non-cervical lymph node metastasis group. Conclusion The positive-expression rate of Notch-2 protein and Numb protein in PTC tissues are higher than those of para-cancerous tissues, and there is a positive correlation between them in PTC tissues, suggesting that there may be a synergistic effect in the course of PTC progression.