【Abstract】 Objective To explore a method to identify the sensory and motor fascicles in peri pheral nervetrunk. Methods Thirty Wistar rats were selected to obtain whole spine. The spinal gangl ion, its dorsal root and ventral root,and sciatic nerve were harvested, Annexin V and Agrin specificities were observed with Western blot. In the experimental group,anterior branch and posterior branch of spinal nerve, sciatic nerve, and its muscular branch and cutaneous branch were harvested from 15 rats to make the observation of immunohistochemistry. In the other 15 rats, first antibody was replaced by PBS as control group. Different nerve fascicles were studied with Micro Raman scattering technique in 16 12-month-old New Zealand rabbits. Results The Annexin V and Agrin were special substances of sensory and motor nerves respectively and can act as specific antigens for identifying different nerve fascicles. There were significant differences in the intensity and breadth of the peak of the spectral properties between motor and sensory fascicles at frequencies of 1 088, 1 276, 1 439, 1 579 and 1 659 cm-1 .The peak intensity ratios of 1 276 to 1 439 cm-1 were 0.95±0.06 in motor nerve fascicles and 1.17±0.08 in sensory fascicles, showing significant differences (P lt; 0.05). Conclusion The Micro Raman spectra is more effective than immunohistochemistry in identifying different nerve fascicles, and it possesses as feasibil ity for cl inical appl ication.
Objective To investigate the expression of cyclooxygenase-2 (COX-2) in liver tissue of severe acute pancreatitis (SAP) rats. Methods A rat model of SAP was induced by retrograde infusion of 3.5% sodium taurocholate into the biliary-pancreatic duct. Eighty rats were randomly divided into SAP group and control group. The levels of serum amylase (AMY), alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor (TNF) -α and ascites AMY were detected dynamically at 4 h, 8 h, 16 h and 24 h after operation. The pancreatic and liver injuries were observed by light microscope. The expression of COX-2 in liver tissue was measured by immunohistochemistry. Results Compared with the control group, the levels of serum AMY, ALT, AST, TNF-α and ascites AMY increased significantly at the time point of 4 h, 8 h, 16 h and 24 h (Plt;0.05). These changes were paralleled with the histopathological changes of pancreatic and liver tissue. The expression positive rates of COX-2 at the different time in the SAP group were higer than those of the control group (Plt;0.05). There was a significantly positive correlation between the expression of COX-2 and ALT (rs=0.949, P=0.039), AST (rs=0.972, P=0.016) and serum AMY (rs=0.944, P=0.041), respectively. Conclusion Overexpression of COX-2 may play an important role in liver injury during SAP.
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.
Objective To establish a stable colorectal cancer model in liver specific insulin-like growth factor (IGF)-1 deficient (LID) mice and examine the potential relationship between IGF-1 level and risk of mice constitutional colorectal cancer. Methods ①Establishment of a colorectal cancer model: The LID mice, in which IGF-1 level in circulation was 25% of BALB/c mice. Induction of colorectal cancer was achieved by using the 1,1 Dimethylhydrazine (DMH) with hypodermic injection at transverse part. ②Eighty fresh samples of cancer tissues and adjacent tissues were obtained from LID mice (experimental group) and BALB/c mice (control group). The expression of IGF-1 was studied by immunohistochemical assay (SP method). Results ①Weight loss occurred in both experimental group and control group after injection. Compared with the body weight before injection on 18 weeks and 24 weeks in each group, there were significant differences after injection at the same phase in each group (P＜0.05). ②The results of IGF-1 expression in cancer tissues and adjacent tissues: IGF-1 got a diffuse distribution in cancer cell cytoplasm. The positive expressions of IGF-1 in the cancer tissues and their adjacent cancer tissues were 6/7, 2/7 and 13/16, 7/16 respectively in experimental group and control group. There were significant differences between the cancer tissues and adjacent tissues inside both groups (P＜0.05). There were no significant differences inside both of cancer tissues and adjacent tissues respectively between experimental group and control group (Pgt;0.05). Conclusion In the established colorectal cancer model by DMH, IGF-1 plays an important role in the development and progression of colorectal cancer.
Objective To investigate the expression and clinical significance of connective tissue growth factor (CTGF) in colorectal cancer. Methods The expressions of CTGF in 62 patients’ colorectal cancer tissues and their corresponding adjacent tissues were detected by SP immunohistochemical method. The results were statistically analyzed. Results The positive rates of CTGF in colorectal cancer and adjacent tissues were 61.3% and 19.4% respectively， and the difference between them was significant (P＜0.05). The expression of CTGF was related to degree of differentiation, depth of infiltration and lymph node metastasis or not (P＜0.05）, namely the lower degree of differentiation, the deeper depth of infiltration and the more lymph node metastasis, the corresponding positive expression rates were lower (P=0.030, P=0.032 and P=0.017 respectively), but correlation with gender was not significant (Pgt;0.05）. Conclusion CTGF may play an important role in the occurrence of colorectal cancer， which contributes a lot to guide clinical treatment and prognosis.
ObjectiveTo investigate the early diagnostic value of transforming growth factor-β1(TGF-β1) on acute rejection after liver transplantation in rhesus by detecting the expression of TGF-β1 in the liver tissue. MethodsLiver transplantation models in rhesus were constructed by the improved vascular dual cuff, supporting tube of biliary tract, and artery anastomosis method.The successful models were randomly divided into experimental group (no immunosuppressant treatment in perioperative period) and control group (treated by immunosuppressant in perioperative period).Then the blood samples and liver tissues were collected at 6, 12, 24, and 72 hours after surgery.Allograft rejections of liver tissue after liver transplantation were monitored by liver function test, hematoxylin-eosin staining and Banff score.Finally, the expression level of TGF-β1 was detected by Western blot analysis or immunohistochemistry technique. Results①The acute rejection happened in all the rhesus at 12 h, 24 h and 72 h after liver transplantation, especially at 72 h after liver transplantation in the experimental group, the Banff grade levels of acute rejection in the liver tissue was more severe than that in the control group (P < 0.05).②The levels of ALT, AST, and TBIL after liver transplantation was gradually increased, which were similar at 6 h and 12 h after transplantation between the two groups, but which at 24 h and 72 h after transplantation in the experimental group were significantly higher than those in the control group (P < 0.05).③The results of TGF-β1 protein expression using immunohistochemical detection:The percentage of positive area of TGF-β1 of liver tissue at 12 h in the experimental group was significantly higher than that in the control group (P < 0.05).With the extension of time, it was gradually increased and significantly higher than that in the control group at 24 h or 72 h (P < 0.05).④The semi-quantitative results of TGF-β1 protein expression using Western blot detection:The TGF-β1 protein expressions began to increase at 6 h after liver transplantation in the experimental group and the control group, and the magnitude of increase was more obvious in the experimental group.The TGF-β1 protein expressions at different time (6 h, 12 h, 24 h, and 72 h) in the experimental group were significantly higher than those in the control group (P value was 0.003, 0.001, 0.001, and 0.001, respectively). ConclusionsThe elevated level of TGF-β1 of liver tissue after liver transplantation might suggest the enhanced cellular immune function, it might have certain significance for early diagnosis of acute rejection after liver transplantation.
ObjectiveTo detect expressions of PTEN and Ki-67 in primary thyroid cancer tissues and explore its clinical significances. MethodsThe expressions of PTEN protein and Ki-67 protein in 40 cases of paraffin-embedded tissues of primary thyroid cancer and the corresponding paracancerous tissues were detected by immunohistochemical method. The expressions of PTEN mRNA and Ki-67 mRNA in 14 cases of resected fresh tissues of primary thyroid cancer and the corresponding paracancerous tissues were detected by RT-PCR method. The relations between clinicopathologic characteristics and expression of PTEN protein or Ki-67 protein in the primary thyroid cancer tissues were analyzed. Results① The PTEN protein positive expression rate and the PTEN mRNA in the primary thyroid cancer tissues were significantly lower than those in the corresponding paracancerous tissues[35.0% (14/40) versus 60.0% (24/40), P<0.05; 0.225 7±0.036 3 versus 0.503 6±0.037 5, P<0.05], the Ki-67 protein positive expression rate and Ki-67 mRNA in the primary thyroid cancer tissues were significantly higher than those in the corresponding paracancerous tissues [72.5% (29/40) versus 42.5% (17/40), P<0.05; 1.212 1±0.042 1 versus 0.293 6±0.027 4, P<0.05]. ② The expressions of PTEN protein and Ki-67 protein were associated with the histological grading, pathological type, tumor stage, and presence of regional lymph node metastasis (P<0.05), which not associated with the patient's gender, age and integrity of tumor capsule or not (P>0.05). ③ The PTEN and Ki-67 protein expressions in the primary thyroid cancer tissues had a significantly negative correlation (rs=-0.605, P=0.000), which in the corresponding paracancerous tissues had no correlation (rs=-0.021, P=0.899). ConclusionPTEN and Ki-67 genes abnormally express in thyroid cancer tissue, which might be related with occurrence and development and its mechanism of primary thyroid cancer. Combination of two genes might contribute to identification of pathologic type, judge of biological behavior, and tumor stage of primary thyroid cancer, which might serve as a new target for diagnosis and treatment of it.
ObjectiveTo investigate the correlation between expression of stromal interaction molecule 1 (STIM1) and tumor malignant degree or lymph node metastasis in patients with gastric cancer. MethodsA total of 83 patients with gastric cancer treated in the Affiliated Hospital of Southwest Medical University and Sichuan Mianyang 404 Hospital from October 2018 to April 2021 were collected. The expression of STIM1 protein in the gastric cancer tissues and the corresponding adjacent normal gastric tissues was detected by immunohistochemistry method. Meanwhile the correlation between the expression of STIM1 protein and clinicopathologic features or postoperative lymph node status of the patients with gastric cancer was analyzed. ResultsThe positive rate of STIM1 protein expression in the gastric cancer tissues was 95.2% (79/83), including 62 (74.7%) patients with high expression (STIM1 scoring 5–7) and 21 (25.3%) patients with low expression (STIM1 scoring 2–4), which in the corresponding adjacent normal gastric tissues was 41.0% (34/83), the difference was statistically significant (χ2=58.078, P<0.001). The expression of STIM1 protein was not related to gender, age, and tumor size of the patients with gastric cancer (P>0.05), while the proportions of the patients with high expression of STIM1 protein in the gastric cancer patients with low/undifferentiated tumor, T3+T4 of infiltration depth, TNM stage Ⅲ, and lymph node metastasis were higher than those with high/medium differentiation (χ2=11.052, P=0.001), T1+T2 of infiltration depth (χ2=24.720, P<0.001), TNM stage Ⅰ+Ⅱ (χ2=9.980, P=0.002), and non-lymph node metastasis (χ2=6.097, P=0.014). The expression intensity of STIM1 protein was positively correlated with the number of lymph node metastasis (r=0.552, Z=–3.098, P=0.002) and the rate of lymph node metastasis (r=0.561, Z=–6.387, P<0.001). ConclusionsPositive rate of STIM1 protein expression in gastric cancer tissues is relatively high. STIM1 protein expression in gastric cancer tissue is closely related to tumor malignancy and lymph node metastasis, so it might play an important role in progression of gastric cancer.