Objective To explore the expression of tumor necrosis factor (TNF) mRNA, TNF and TNFR in the gallbladder mucosa which developed from hyperplasia, dysplasia to carcinoma, and to further discuss the relationship between TNF and pathogenesis of gallbladder carcinoma. Methods In situ hybridization and immunohistochemistry were used to determine TNF mRNA, TNF protein and TNFR protein expression in hyperplasia, dysplasia and carcinoma of gallbladder. Results ①No one of 20 cases of gallbladder hyperplasia was found to express TNF mRNA, while 4 of 20 (20%) cases of dysplasia and 18 of 20 (90%) cases of carcinoma were found to express TNF mRNA (P＜0.05). ②For the expression of TNF mRNA in mononuclear cells (MNC), positive staining was found in 15% of gallbladder hyperplasia, 85% of dysplasia and 90% of carcinoma, respectively (P＜0.05). The cell numbers of positive staining MNC were 4.85±1.50, 6.00±2.71 and 9.33±3.07, respectively (P＜0.05). ③In gallbladder carcinoma, the cell number of carcinoma and MNC with positive TNF mRNA expression was correlated with clinical stage (P＜0.05). The higher the clinical stage, the more the positive staining cell numbers. The positive staining cell numbers of carcinoma in stage Ⅰ-Ⅲ and Ⅳ-Ⅴ were 9.13±4.39 and 14.80±4.02, respectively (P＜0.01), and the positive staining cell numbers of MNC were 7.13±2.53 and 11.10±2.23, respectively (P＜0.05). ④The cell numbers of carcinoma and MNC with TNF mRNA expression increased with tumor size. In tumors with diameter over 2 cm and less than 2 cm, the positive staining cell numbers of carcinoma were 14.00±4.20 and 8.83±4.96, respectively (P＜0.05), and that of MNC were 10.50±2.54 and 7.00±2.83, respectively (P＜0.05). ⑤The region of TNF protein expression was similar to that of TNF mRNA, but TNF protein expression was more frequent and wider than that of TNF mRNA. ⑥The tumor necrosis factor receptor was expressed in tumoral vascular endothelial cells and MNC in all cases of carcinoma, but was negatively stained in mucosa epithelial cells and tumor cells of all cases. ⑦There was positive linear correlation in TNF mRNA between tumor cell and MNC (r=0.687, P＜0.01), same as that in TNF protein expression (r=0.742, P＜0.01); and there was positive linear correlation in tumor cell between TNF mRNA and TNF protein expression (r=0.847, P＜0.01), same as that in MNC (r=0.643, P＜0.01). Conclusion The TNF mRNA and TNF protein expression are increasing during the development of gallbladder mucosa epithelial from hyperplasia, dysplasia to carcinoma, and increasing with tumor stage. It suggests that TNF may contribute to carcinogenesis of gallbladder carcinoma induced by gallstone, and related to the progression of gallbladder carcinoma.
ObjectiveTo study the expression of HOX A9 mRNA and its clinicopathological significance in the benign and malignant lesions of pancreas. MethodsIn situ hybridization for HOX A9 mRNA was used on routine paraffinembedded sections. ResultsThe positive rate and scoring mean of HOX A9 mRNA expression was significanfly lower in pancreatic carcinoma (49%, 3.3±2.1) than that in chronic pancreatitis (95%, 5.4±0.8) and pericancerous tissues (80%, 4.6±1.2), the negative case of HOX A9 mRNA in chronic pancreatitis and pericancerous tissues showed middle or severelyatypical hyperplasis of the ductal epitheli. The positive rate and scoring mean of HOX A9 mRNA expression was significantly higher in the cases of welldifferentiation (63%, 4.0±2.2) or without metastasis (64%, 4.1±2.2) than that in the ones of poorlydifferentiation (32%, 2.6±2.3) or with metastasis (32%, 2.7±2.2). ConclusionThe expression of HOX A9 mRNA might be related the carcinogenesis, progress, biological behaviors, and prognosis of pancreatic carcinoma. The assay of HOX A9 mRNA expression in the benign lesions of pancreas might have important clinical values in the prevention and earlystage finding of the pancreatic carcinoma.
【Abstract】Objective To study the relationship of the expression of CD44v6 mRNA and nm23H1 mRNA with the clinical pathology parameter and prognosis of breast cancer, and to investigate the correlation of the expression of CD44v6 mRNA and nm23H1 mRNA. Methods In situ hybridization and CSA immunohistochemistry method were used to detect the expression of CD44v6 mRNA and nm23H1 mRNA in 94 cases of breast cancer. Results The positive expression of CD44v6 mRNA and the negative expression of nm23H1 mRNA were positively correlated with the grading, clinical stage, lymph node metastasis, and recurrence and prognosis of breast cancer. CD44v6 mRNA expression and nm23H1 mRNA were negatively correlated in breast cancer. Patients who had positive expression of CD44v6 mRNA and negative expression of nm23H1 mRNA had a higher lymph node metastatic rate and a lower survival rate. Conclusion Several genes were involved in the occurrence and development of breast cancer in which the expression of CD44v6 mRNA has synergistic action in negative regulation with that of nm23H1 mRNA. Combined detection of the expression of these two mRNA is helpful to judge the metastasis, recurrence and prognosis of breast cancer.
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.
Effect of radical operation on expression of interleukin-2(IL-2)mRNA and production of IL-2 were markedly reduced preoperatively and four weeks after operation,expression of IL-2 mRNA significantly enhanced,but it was still lower than that in the normal group.Production of IL-2 nearly reached normal level,When PBL was activated by phytohemagglutinin(PHA),expresseion of IL-2 mRNA and production of IL-2 were much higher than that in non-activated PBL.These results suggested that expression of IL-2 were much higher than that in non-activated PBL.These results suggested that expression of IL-2 mRNA and production of IL-2 are dificient in gastric cancer patients,and radical surgery will help them to recover and they can also be improved through activation with PHA.
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 changes in the expression level of PDGF in the bone callus of rats with femoral fracture and brain injury to explore the effect of brain injury on the fracture heal ing and the related mechanism. Methods Sixty-four 12-week-old SD rats weighing (356 ± 25) g were randomly divided into 8 groups with 8 rats in each. The rats in groups A1, B1, C1 and D1 had a femoral fracture and a brain injury for 1, 2, 3 and 4 weeks, respectively; the rats in groups A2, B2, C2 and D2 had a mere fracture without a brain injury for 1, 2, 3 and 4 weeks, respectively. After the CR films were taken, the bone callus was obtained 1, 2, 3 and 4 weeks after operation, respectively. Then, the bone callus and its histology were examined by HE staining, the expressions and changes in the level of PDGF were examined by the immunohistochemical staining, and the level of PDGF mRNA was measured by in situ hybridization. Results The CR films showed that the callus formation in the A1-D1 groups was earl ier and greater than that in the A2-D2 groups at the same time point. The HE staining indicated that more fibroblasts and early-stage chondrocytes were found in group A1; some fibroblasts in the fracture interspace and few early-stage chondrocytes were found in group A2; some newly-formed trabecular bones were found at the end of the fracture in group B1; but no trabecular bone formation was found in group B2; woven bone formation and a few chondrocytes between trabecular bones in the fracture interspace were found in group C1; only a few trabecular bones in the fracture interspace were found in group C2；woven bones turned to lamellar bones in group D1；and more immature trabecular bones in the fracture interspace were found in group D2. The positive expression of PDGF and PDGF mRNA was b in the cytoplasms of fibroblasts, mesenchymal cells, vascular endothel ial cells, early-stage chondrocytes, osteoblasts and osteoclasts. The percentage of the positive cells for PDGF and PDGF mRNA in the callus was significantly higher in groups A1-D1 than in groups A2-D2 at the same time point (P lt; 0.05). Conclusion Brain injury can promote the fracture heal ing process, which is probably related to an increase in the expression level of PDGF after the brain injury.
Objective To research the gene expression of transforming growth factor β1 (TGF-β1) in zone Ⅱ flexor tendon wound healing of rabbit. Methods Sixty New Zealand white rabbits forepaws(left side) underwent complete transection and the middle digit flexor digitorum profundus tendon in zone Ⅱ were repairedby Kessler methods as the experimental group. The normal right forepaws served as the control group. The tendons and tendon sheaths were harvested at 1, 7, 14, 21, 28and 56 days after repair(n=10). The expression patterns ofTGF-β1 wereanalyzed by in situ hybridization and immunohistochemistry staining methods. Results The in situ hybridization examination revealed thatTGF-β1 mRNA expression upregulated at 1 day, reached the peak levels at 1421 days and remained high levels up to 56 days in the experimental group. The expression ofTGF-β1 mRNA in control group was lowerthan that in the experimental group, showing statistically significant difference (Plt;0.05). The results of immunohistochemical staining was similar to that of in situ hybridization. Conclusion The normal tendon and tendon sheath cells are capable ofTGF-β1 production. The cytokine is activated in tendon wound condition. The upregulation of this cytokine in both tendon and tendon sheath cells are coincidence with both extrinsic and intrinsic mechanisms for tendonrepair.
Objective To compare gene express difference ofkeloid and normal skin tissues by using the suppression subtractive hybridization (SSH) so asto find the differential express gene in keloid. Methods mRNA extracted fromkeloid and normal skin tissues was used as the template to synthesis cDNA of keoid and normal skin. The cDNA of keloid served as a tester, the cDNA of normal skin as a driver. cDNA was digested with RsaⅠ. Adaptor-ligated tester cDNA was prepared. Then first hybridization, second hybridization and PCR amplificationwere done. Differentially expressed cDNA was selectively amplified during thesereactions. After SSH, the PCR mixture was ligated with T-vector. The positive clones were selected and the insert gene fragments were analyzed. Southern hybridization identified the keloid differential express genes. The positive clones ofSouthern hybridization were selected, and these sequences were analyzed. The results were compared with that of GeneBank. Results Thirteen differential genes were found in keloid, of which 11 gene clones have been known their function, and 2 clones have not known their function. 〖WTHZ〗Conclusion Keloid differentially expressed gene was screened successfully by SSH.
Objective To study the mRNA expressions of protein kinase C(PKC) and nerve growth factor (NGF) in rat sciatic nerve and the number ofaxons after phorbol-12-myristate-13-acetate (PMA) was injected into silicone chamber. Methods Forty-two SD adult rats were divided into six groups depending on the time of injury (1 day, 3 days, 1 week, 2 weeks, 3 weeks and 4 weeks). A 0.5 cm nerve was cut in doublerat sciatic nerves and “T” type silicone chamber was sutured. PMA at the concentration of 1×10-9mol/L was injected discontinuously into the right side of Ttype silicone chamber(PMA group) and saline was injected into the left side(control group). Nucleic acid in situ hybridization histochemistry technique and thecomputer imagine analysis were employed to detect dynamic changes of PKC mRNA and NGF mRNA in rat sciatic nerves. The number of axons was measured. Results The expressions of PKC mRNA and NGF mRNA increased after injury, and the expressions of PKC mRNA and NGF mRNA reached the peak 2 weeks and3 weeks after injury respectively in control group. The expressions of PKC mRNA and NGF mRNA in PMA group were significantly increased than those in control group 2,3 and 4 weeks after injury(Plt;0.01).The number of axons in PMA group significantly increased than that in control group(Plt;0.01). Conclusion PKC involved inthe expression of NGF mRNA and nerve regeneration after injury. During the regenerated course, PMA can promote the expression of NGF mRNA and the number of axons after injury.