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.
ObjectiveTo study the clinicopathological features of mediastinum nodular sclerosis Hodgkin lymphoma (NSHL) in order to improve the recognition of it.
MethodsThe clinical data of 3 cases of mediastinum NSHL between 2003 and 2012 were collected. Then we analyzed the carcinoma pathologic samples by pathomorphology, immunophenotypic phenotype, related gene rearrangement and situ hybridization with EBER.
ResultsThe pathomorphologic results showed that broad fibrotic bands subdivided the lymphoid parenchyma into large nodules, the tumoral cells had distinct boundary with empty cytoplasm and small-to-medium-sized nucleoli, and the nodules contained inflammatory cell components. The immunophenotypic phenotype of the tumoral cells were CD15, CD30, PAX-5 and CD20 partly, but anaplastic lymphoma kinase, CD45, cytokeratin, CD79α and S-100 were not expressed. T cell receptor γ and IgH gene were no rearranged, and EBER in situ hybridization was not detected.
ConclusionVarious lymphomas occur in the mediastinum and mediastinum NSHL is just one of them. Mastering its distinctive pathomorphology and immunophenotypic phenotype is highly significant for diagnosis, differential diagnosis and treatment of the disease.
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.
For observation of the change of transforming growth factor-beta 1 (TGF-beta 1) gene expression in the process of skin wound healing, the following experiments were performed. Sixteen Wistar rats were chosen. At each side of the rat’s back, a 1 cm x 1.5 cm middle-thick skin wound was made. After 3, 6, 9 and 12 days, the specimens were taken from the wounds. For each specimen, half of it was used for RNA extraction, and underwent dot blotting; and the other half was frozen immediately and underwent in situ hybridization. The probes were dig-labeled PDGF-BB cDNA probe and TGF-beta 1 probe. The results showed that TGF-beta 1 gene was expressed mainly in fibroblast, epithelial cell and capillary endothelial cell. The peak of TGF-beta 1 mRNA content was in the 6th day postoperatively. After that, the content of TGF-beta 1 decreased to normal. It was suggested that TGF-beta 1 gene expression was in close relation with healing process. TGF-beta 1 may play an important regulatory role in the skin wound healing.
Objective
To investigate expressions of EphA2 and EphrinA1 in invasive ductal carcinoma of breast and to explore their clinical significances.
Method
The protein and mRNA expressions of EphA2 and EphrinA1 in 30 breast fibroma tissues, 30 breast cystic hyperplasia tissues, and 100 invasive ductal carcinoma of breast tissues were detected by immunohistochemistry andin situ hybridization respectively, and correlation between them and relations between their expressions in invasive ductal carcinoma of breast tissues and clinicopathologic factors were analyzed.
Results
① The results of the immunohistochemistry andin situ hybridization tests showed that the protein and mRNA expressions of EphA2 and EphrinA1 in the invasive ductal carcinoma of breast tissues were significantly higher than those in the breast fibroma tissue (P<0.001) and breast cystic hyperplasia tissue (P<0.001). ② The positive expressions of EphA2 and EphrinA1 protein and mRNA were associated with the lymph node metastasis, histological grade, and TNM stage (P<0.05), in other words, which in the invasive ductal carcinoma of breast patients with lymph node metastasis, high histological grade, and high TNM stage were higher. However, which were not associated with the age and the tumor diameter (P>0.05). ③ The positive protein expressions or positive mRNA expressions in the invasive ductal carcinoma of breast tissues all had positive correlations between the EphA2 and the EphrinA1 (protein:rs
=0.999,P<0.01; mRNA:rs
=0.942,P<0.01).
Conclusions
EphA2 and EphrinA1 might be involved in carcinogenesis and development procedures of invasive ductal carcinoma of breast. Combined detection of EphA2 and EphrinA1 could help to predict clinical and pathologic characteristics of invasive ductal carcinoma of breast. They might provide a new target for clinical medication, prognosis, and targeted therapy.
【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 influnce of L-arginine (L-Arg) and L-nitro-arginine-methyl-ester(L-NAME) to purified retinal ganglion cells(RGCs) apoptosis of rats cultured in different consistencies of L-Arg and L-NAME.
Method
RGCs from Sprague Dawley (SD) neonatal rats(postnatal 1~5 day) were cultured in assimilative culture solution in vitro and RGCs were purified by Thy1.1 with sheep anti rat FITC monoclonal antibody. RGCs were cultured in different consistencies of L-Arg and L-NAME: 1×10-6, 1×10-5,1×10-4, 1×10-3, 1×10-2 and 1×10-1 mol/L for 24 hours and 48 hours, respectively. The changes of bcl-2, bax and p53 mRNA in RGCs in different consistencies of L-Arg and L-NAME
were demonstrated qualitatively and quantitatively by in situ hybridization, and their apoptosis were detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL) method, respectively.
Results
After 24 hours in vitro, the purification rate of RGCs in the experiment arrived at 97 %. After 48 hours, there were a few apoptotic cells expression in the control group. Apoptotic cells expression in L-Arg≥1×10-3 mol/L and L-NAME≥1×10-1 mol/L groups increased that had a significant difference with the control group (Plt;0.05). In the group of L-Arg≥1×10-3 mol/L and L-NAME≥1×10-1 mol/L, the expression of bcl-2 mRNA in RGCs became weaker and weaker as the consistencies were increased, but the expression of bax and p53 mRNA in RGCs became higher and higher and had a significant difference with control group (Plt;0.05).
Conclusion
Lower concentration of L-Arg can promote the growth of purified RGCs in vitro and higher concentration of L-Arg can promote the apoptosis of RGCs.
(Chin J Ocul Fundus Dis, 2002, 18: 137-139)
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.
Objective To investigate the relationships between circulating tumor cells (CTCs), circulating tumor endothelial cells (CTECs) and treatment methods in patients with nasopharyngeal carcinoma (NPC) at different stages of treatment. Methods The data of NPC patients at different treatment periods in West China Hospital of Sichuan University from March 2016 to November 2019 were retrospectively collected. The patients received CTCs test and part of those patients received CTECs test, by subtraction enrichment-immunostaining-fluorescence in situ hybridization. The relationships of CTCs and CTECs with radiotherapy and chemotherapy, and the correlations between CTCs and CTECs in NPC patients were analyzed. Results A total of 191 patients were included. Among them, there were 66 cases before initial treatment, 38 cases after induction chemotherapy, and 87 cases after concurrent chemoradiotherapy. A total of 127 patients received CTECs test, including 41 cases before initial treatment, 29 cases after induction chemotherapy, and 57 cases after concurrent chemoradiotherapy. The positive rates of CTCs were 89.4%, 81.6% and 69.0% respectively in the three stages of treatment, and the difference was statistically significant only between the pre-treatment group and the post-concurrent chemoradiotherapy group (P=0.003). The number of CTCs in the post-concurrent chemoradiotherapy group was lower than that in the pre-treatment group and the post-induction chemotherapy group (P<0.001, P=0.002). The number of triploid CTCs in the post-concurrent chemoradiotherapy group was significantly different from that in the pre-treatment group and the post-induction chemotherapy group (P=0.009, P=0.013). The number of tetraploid CTCs in the post-concurrent chemoradiotherapy group was significantly different from that in the post-induction chemotherapy group (P=0.007). The number of polyploidy (pentaploid or > 5 copies of chromosome 8) CTCs in the post-concurrent chemoradiotherapy group was significantly different from that in the pre-treatment group (P<0.001). The positive rates of CTECs were 70.7%, 82.8% and 64.9% respectively in the three stages of treatment, and the difference was not statistically significant (P>0.05). The number of CTECs in the post-concurrent chemoradiotherapy group was only lower than that in the post-induction chemotherapy group (P=0.009). There was no significant difference in the number of triploid or tetraploid CTECs among the three groups (P=0.265, P=0.088). The number of polyploid CTECs was statistically different only between the post-concurrent chemoradiotherapy group and the post-induction chemotherapy group (P=0.007). Spearman correlation analysis showed that there was a significant positive correlation between CTCs and CTECs (rs=0.437, P<0.001). Conclusions Concurrent chemoradiotherapy plays a decisive role in reducing the number of CTCs in the blood of NPC patients, while induction chemotherapy does not appear to directly cause changes in the number of CTCs. In NPC patients, different types of CTCs have different responses to different treatments. There is a significant positive correlation between CTECs level and CTCs level in NPC.
ObjectiveTo investigate the expressions of Snail and VEGF gene in invasion ductal carcinoma tissues and analyze their clinicopathologic relationship. MethodsThe expressions of Snail and VEGF gene were detected on mammary gland hyperplasia (30 cases), intraductal breast cancer (30 cases), and invasion ductal carcinoma (70 cases) by in situ hybridization， to compare with the expression difference of the two genes in the different pathological changed tissues of mammary gland and among the clinicopathological facters of invasion ductal carcinoma as well as the relationship. ResultsThe expression rate of Snai mRNA in mammary gland hyperplasia， intraductal breast cancer, and invasion ductal carcinoma was 23.3% (7/30)， 46.7% (14/30), and 81.4% (57/70), respectively, there was statistical difference among them (χ 2=32.4, Plt;0.05)； The expression rate of VEGF mRNA in mammary gland hyperplasia， intraductal breast cancer, and invasion ductal carcinoma was 33.3% (10/30)， 50.0% (15/30), and 71.4% (50/70), respectively, there was statistical difference among them (χ 2=13.4, Plt;0.05). The expression rates of Snail mRNA and VEGF mRNA in lymphatic metatasis group were significantly higher than those in no lymphatic metatasis group 〔92.7% (38/41) vs. 65.5% (19/29)， χ 2=8.29, Plt;0.05； 85.4% (35/41) vs. 51.7% (15/29), χ 2=9.42, Plt;0.05， respectively 〕. The expression rates of Snail mRNA and VEGF mRNA in Ⅲ-Ⅳ stage of TNM clinical stage were significantly higher than those in Ⅰ-Ⅱ stage 〔939% (46/49) vs. 52.4% (11/21), χ 2=14.14， Plt;0.05； 81.6% (40/49) vs. 47.6% (10/21), χ 2=8.32， Plt;0.05〕. The expressions of Snail mRNA and VEGF mRNA were related to the expressions of ER, PR, HER-2, and vessel cancer embolus （Plt;0.01）. The expressions of Snail mRNA and VEGF mRNA were not related to age, tumor size, and histological grade (Pgt;0.05). There was a positive correlation between the expressions of Snail mRNA and VEGF mRNA （r=0.67， Plt;0.05）. ConclusionsThe overexpressions of Snail mRNA and VEGF mRNA in invasion ductal carcinoma has a synergetic effect on occurrence and development, therefore, combined detecting the expressions of Snail mRNA and VEGF mRNA are some significance to predict infiltration and metastasis of the invasion ductal carcinoma.
