ObjectiveTo investigate the expression and relation of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in rats with diabetic retinopathy.MethodFifty-five Wistar rats were randomly divided into the control group (10 rats), and 1, 3, and 5-month-diabetes group (15 rats in each diabetes group), and the diabetic models were set up. The expressions of VEGF and bFGF were detected by situ hybridation and immunohistochemistry on retinal paraffin sections.ResultsThe results of situ hybridation showed that expression of bFGF was found in 3-month-deatbtes group with the percentage of 77.8%, and 88.9% in 5-month-deatbtes group; the positive expression of VEGF was not found in 3-month-deatbtes group but in 5-month-deatbtes group with the percentage of 66.7%. Immunohistochemistry indicated that the positive expression of bFGF started in 3-month-deatbtes group with the percentage of 55.6%, and 88.9% in 5-month-deatbtes group; the percentage of the expression of VEGF was 33.3% in 3-month-deatbtes group and 88.9% in 5-month-deatbtes group.ConclusionThe expression of VEGF occurs after the expression of bFGF in rats with DR.(Chin J Ocul Fundus Dis, 2005,21:37-40)
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.
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.
Objective To identify the expression of pleiotrophin （PTN） mRNA and protein in the colorectal cancer tissues， and to explore the clinical value of it. Methods The expressions of PTN mRNA and protein in colorectal cancer tissues （colorectal cancer group） as well as normal colorectal tissues （normal control group） were tested by using in-situ hybridization and immunohistochemistry. Results The positive rates of PTN mRNA 〔63.9% （53/83） vs. 40.7%（22/54）〕 and protein〔60.2%（50/83） vs. 33.3%（18/54）〕 in colorectal cancer group were all significantly higher than those of normal control group （P=0.008， P=0.002）. There were no significant relationship between expressions of PTN mRNA and protein with gender， age， and type of tumor （P＞0.05）， but in tissues of Ⅲ-Ⅳ stage and poor differentiation，the positive rates of PTN mRNA and protein were all higher （P＜0.05）. Conclusions The over expressions of PTN mRNA and protein in colorectal cancer tissues may directly related to the invasion and metastasis. Meanwhile， it can be used as an index to predict metastasis and prognosis of colorectal cancer.
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.
To investigate the function of nitric oxide (NO) and nitric oxide synthetase (NOS) inhibitor, N-nitro-L-arginine methyl ester (L-NAME), the skin avulsion model was made in the lower extremity of pig. The methods of measurement of size of the survived flap, weighing, immunocytochemistry and hybridization in situ were employed, so that the survival surface area of flaps, tissue wet/dry weight ratio, NO content in the serum, gene expression of NO and NOS content in the flap tissue were determined, respectively. The results showed that the early gene expression of NOS was increased as well as the NO content and tissue wet/dry weight ratio (P lt; 0.01). After L-NAME was applied introvenously, the NO content and tissue wet/dry weight ratio were decreased (P lt; 0.01), and the survival surface area of flaps was enlarged (P lt; 0.01). It could be concluded that the NO might play a role in the development of the pathological changes as early congestion, edema and secondary necrosis in the avulsed skin flaps. The early application of L-NAME could do some good to the avulsed skin flap and protect it from further necrosis owing to the presence of NO.
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.