ObjectiveTo investigate the expression of caspase-3 and Toll-like receptor 4 (TLR4) in the incised rat skin healing process and its relationship with the wound time and to provide an experimental evidence for the prediction of injury time. MethodsAfter the rat incised wound model was established, hematoxylin-eosin dyeing technology and immunohistochemical staining technique were used to observe the expression of caspase-3 and TLR4. Then Image Pro Plus Image analysis software and SPSS statistical analysis software were used to deal with the experimental results. ResultsCaspase-3- and TLR4-positive cells were detected in epidermis, hair follicle and sebaceous gland cells in the control skin. The expression of caspase-3 and TLR4 of the ante mortem groups were significantly different compared with the control group except the 0 h group (P<0.05). Caspase-3- and TLR4-positive cells were detected in neutrophils around the hair follicle half an hour later. Caspase-3- and TLR4-positive cell rate increased with the infiltration of inflammatory cells. Caspase-3- and TLR4-positive cell rate reached the maximum on the 3 rd day, and then it began to decrease, and they were mainly expressed in fibroblasts and mononuclear macrophages. Caspase-3- and TLR4-positive cells were mainly expressed in fibroblasts on the 10th day. There was no significant differences between the postmortem injury groups and the normal control groups (P>0.05). ConclusionCaspase-3- and TLR4-positive cell rate is time dependent and stable in 25℃ temperature environment which makes it possible to determine the time of injury.
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 prevalence of cellular FLICE-like inhibitory protein (cFLIP) alterations in colorectal cancer and their possible implications for the progression of colorectal cancer. Methods The long type cFLIP (cFLIPL) was examined in 43 colorectal cancer specimens and the matched normal colorectal specimens by immunohistochemistry. Immunohistochemical staining for cFLIPL was assessed on an arbitrary semi-quantitative scoring system. Stained cells were counted under the magnification field (×100) and at least 20 fields per case were examined. Fields with non-stained cells were scored 0; fields with stained cells less than 5% were scored 1; fields with stained cells from 5% to 25% were scored 2; fields with stained cells between 26% and 50% were scored 3; and fields with stained cells more than 50% were scored 4. According to the above schedule， a mean score of each specimen was calculated. Results cFLIPL protein expressions of variable intensity and extent were detected in the normal colorectal mucosa and adenocarcinomas. cFLIPL was mainly expressed in the cytoplasma. The positive cells were distributed in diffuse manner. The mean expression score in normal mucosa ranged from 0 to 2.95 (mean score: 1.55±0.86); 4.7%(2/43) of all cases were unstained and 20.9%(9/43) showed a weakly stained pattern (0＜mean score≤1); 74.4%(32/43) of all cases were positively stained (1.00＜mean score≤2.95), respectively. cFLIPLprotein was expressed in all adenocarcinomas and the score ranged from 0.80-4.00 (mean score: 3.06±0.75); 62.8% (27/43) of the tumors examined were predominantly cFLIPL positive (mean score ＞3), 34.9%(15/43) of the tumors were cFLIPLpositive (1＜mean score ≤3) and only one case was cFLIPL weakly positive (score: 0.80). 72.1% (31/43) of adenocarcinomas expressed cFLIPLmore intensely and extensively than matched normal colonic mucosa. cFLIPL expression of colorectal cancer was significantly higher than that of matched normal mucosa, which was statistically significant (P＜0.01). The extent of cFLIPL expression in tumors was independent of Dukes stage, tumor stage, gross type of tumor, histological type, or lymph and hepatic metastasis. Conclusion The expression level of cFLIPL in adenocarcinomas is much higher than that in matched normal mucosa. Overexpression of cFLIPL is a tumor-specific phenomenon, which can provide a selective growth advantage for colorectal cancer cells to escape from death receptor-mediated apoptosis.
Objective To investigate the role of expression in the differential diagnosis of thyroid follicular carcinoma and follicular variant of papillary carcinoma. Methods Seventy cases of thyroid lesions (including 15 cases of follicular adenomas, 15 cases of adinomatous goiters, 30 cases of papillary carcinomas and 10 cases of follicular carcinomas) were collected, and CD10 expression was detected by means of immunohistochemistry in above thyroid lesions. Results Seven of 9 cases of follicular variant of papillary carcinoma were CD10 positive (77.8%), and 8 of 10 cases of follicular carcinoma were CD10 positive (80.0%). However, CD10 was negative in all cases of non-follicular variant of papillary carcinoma, follicular adenoma, adinomatous goiter and normal thyroid tissue. Conclusion The detection of CD10 expression is useful to the differential diagnosis of thyroid follicular carcinoma and follicular variant of papillary carcinoma.
Objective To detect the expressions of CK5/6 and Ki-67 in breast cancer， and explore the clinical significance. Methods The expressions of CK5/6 and Ki-67 were detected by immunohistochemistry in 162 cases of breast cancer . The correlation between CK5/6 expression and Ki-67 expression and the relationship of the expressions of two factors to the clinicopathologic factors were analyzed. Results There were 12 cases of the triple negative breast cancer 〔negative estrogen receptor （ER）， negative progesterone receptor （PR）， and negative Her-2〕 in 162 patients with breast cancer. The positive rates of CK5/6 and Ki-67 in the breast cancer tissues were 30.9% （50/162） and 65.4% （106/162），respectively. The positive rates of CK5/6 and Ki-67 in the patients with triple negative breast cancer were significantly higher than those of non-triple negative breast cancer 〔CK5/6：75.0% （9/12） versus 27.3% （41/150）， χ2=11.837， P=0.001；Ki-67：100% （12/12） versus 62.7% （94/150）， χ2=6.847， P=0.009〕. The expressions of CK5/6 and Ki-67 in the breast cancer tissues were not associated with age （P＞0.05）， but they were associated with histology grade （P＜0.05）. The expression of CK5/6 wasn’t associated with tumor size and lymph node status （P＞0.05）， but the expression of Ki-67 was associated with them （P＜0.05）. The expression of CK5/6 in the breast cancer tissue had a positive correlation with the expression of Ki-67 in the breast cancer tissue （rs=0.271， P=0.000）. There were 64 cases when the expression of Ki-67 was （＋＋） and （＋＋＋）， the positive rate of CK5/6 was 43.8% （28/64） and it was significantly higher than that when the expression of Ki-67 was （-） and （＋） 〔22.4% （22/98）， χ2=8.233， P=0.004〕. The positive expressions of CK5/6 and Ki-67 in the breast cancer tissues were negatively correlated with the expressions of ER and PR （CK5/6 and ER：rs=-0.446， P=0.000；CK5/6 and PR：rs=-0.370， P=0.000；Ki-67 and ER：rs=-0.518， P=0.000；Ki-67 and PR：rs=-0.515， P=0.000）. The positive expressions of CK5/6 and Ki-67 in the breast cancer tissue were not correlated with the Her-2 expression （CK5/6 and Her-2：rs=0.105， P=0.183；Ki-67 and Her-2：rs=0.068， P=0.393）. Conclusion The expressions of CK5/6 and Ki67 not only can evaluate the basic rules of breast cancer from origin and development， but also they are beneficial to choose the chemotherapy of different patients.
Objective To investigate the expression of ADAM9 in breast cancer and its clinical significance. Methods The expressions of ADAM9 in normal breast tissues and breast cancer tissues were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry, and whose relationship with clinicopathologic features was analyzed. Results The expression of ADAM9 mRNA increased in the breast cancer tissues, but which was not detected in the normal breast tissues. The expression of ADAM9 protein in the breast cancer tissues was significantly higher than that in the normal breast tissues (Plt;0.05), and which in the metastatic lymph nodes was significantly higher than that in the negative lymph nodes or corresponding primary lesions (Plt;0.05). The expression of ADAM9 in the breast cancer tissues was correlated with the lymph node metastasis and histological grade (Plt;0.05). Conclusion ADAM9 is overexpressed in the breast cancer tissues, which might involve in the pathological progression of breast cancer.
Objective To investigate the method of cultivation and the feature of differentiation of spinal cordderived stem cells in vitro.Methods The neural stemcells from spinal cord of 15 days fetal rats were harvested and cultivated in aserumfree limited medium. The stem cells were induced to differentiate and theresults were identified by cellular immunohistochemistry. Results Lots of stem cells were obtained from the spinal cord of fetal rats and the sphere of stemcells was formed about 10 days. Neural stem cells can give rise to mature neurons and astrocytes.Conclusion Epidermal growth factor/basic fibroblast growth factor serum-free limited medium can promote the proliferation activity ofthe stem cells. Spinal cord-derived stem cells can differentiate into glial cells and neurons.
Objective To research the effect of γ-ray released from 103Pd radioactive stent on the expression of Fas gene and its relation with apoptosis of cholangiocarcinoma cell and the significances through the establishment of human cholangiocarcinoma model. Methods The model of nude mouse with implanted human cholangiocarcinoma was established. The mice were divided into study group and control group, 37 MBq 103Pd biliary stent was implanted in the study group and the ordinary metal biliary stent was implanted in the control group. The volume of tumor was measured, the cell apoptosis was detected by the TUNEL method and the expression of Fas gene of the cell apoptosis of the induced human cholangiocarcinoma was checked out by immunohistochemistry staining 10 d after the implantation. Results Compared with the control group, the growing speed of the volume of tumor in study group was significantly reduced (Plt;0.05), the expression positive rate of Fas gene was significantly higher (Plt;0.05), and the apoptotic rate of cancer cells was also higher (Plt;0.01). Conclusions The 103Pd radioactive stent can induce the cell apoptosis in nude mouse model with implanted human cholangiocarcinoma inhibit the cell growth of bile duct cancer and may promote the apoptosis of cancer cells by increasing the expression of Fas gene. It may be helpful for the further study of treatment for bile duct cancer using 103Pd radioactive stent.
ObjectiveTo investigate the expression and distribution of CD15s antigen in breast cancer and its relationship with carcinogenesis, progression and metastatic proclivity. MethodsCatalyzed signal amplification(CSA) immunohistochemical technique was used to detect the expression of CD15s antigen in breast cancer and in adjacent normal mucosa. Immunoelectromicroscopic ultrastructural localization of CD15s antigen labelled by colloidal gold was also bserved.ResultsThe positive rate of CD15s antigen expression in primary breast cancer was 79.8%(75/94). In adjacent normal mucosa (n=10) CD15s antigen showed weaker staining. The positive rate of CD15s antigen expression in grade Ⅱ-Ⅲ (87.3%) was notably higher than that in grade Ⅰ (69.2%, P＜0.05). In patients with lymph node metastasis, the positive rate of CD15s antigen expression was 90.2%, which was significantly higher than 67.4% in nodes with no metastasis (P＜0.05). CD15s antigen immunoreactivity was mainly localized in the border membrane of cytoplasm, endoplasmic reticulum, golgi complex and surrounding nuclear membrane in tumor tissue, and in the border membrane of cytoplasm in adjacent normal tissue. Conclusion CD15s antigen is a practical parameter for evaluating the degree of malignancy and lymphatic metastatic proclivity of breast cancer. It can provide a new pathway to investigate the carcinogenesis and progression of breast 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.