ObjectiveTo investigate the expression level and the clinical significance of Traf-2 and Nck-interacting protein kinase (TNIK) in gastric cancer tissues.MethodsWe retrospectively collected 78 cases of gastric cancer and its adjacent tissues which were diagnosed by surgery and pathology, all the patients received surgery in the Department of General Surgery in Luoyang Central Hospital Affiliated to Zhengzhou University, from July 2014 to December 2017, as well as 42 cases of gastric ulcer tissues in the same period. The expression of TNIK protein was detected by using immunohistochemical method and Western blot method in 78 cases of human gastric cancer tissues, corresponding adjacent normal tissues, and 42 cases of gastric ulcer tissues, the relationship between expression of TNIK protein and clinicopathologial features of gastric cancer was explored.ResultsThe positive expression rate of TNIK protein in gastric cancer tissues was 66.67% (52/78), and adjacent cancer tissues showed low expression rate of 11.54% (9/78), while it was 21.43% (9/42) in gastric ulcer tissues. On the positive expression rate of TNIK protein, gastric cancer tissues compared with adjacent cancer tissues and gastric ulcer tissues, the difference was statistically significant (P<0.05), however, there was no statistically significant difference between adjacent cancer tissues and gastric ulcer tissues (P>0.05). Western blot method showed that the expression level of TNIK protein in gastric cancer tissues was all significantly higher than that in adjacent cancer tissues and gastric ulcer tissues, the differences were statistically significant (P<0.05), but the difference was not significant when compared the adjacent cancer tissues and gastric ulcer tissues (P=0.772). The expression of TNIK protein was irrelevent to sexual distinction, CEA value, tumor diameter, and pathological type (P>0.05), while it was closely related to patients’ age, TNM stage, differentiation grade, distant metastasis, and lymph node metastasis (P<0.05). The prognosis of patients with positive expression of TNIK protein was worse than that of patients with negative expression of TNIK protein (P<0.05).ConclusionTNIK protein is highly expressed in gastric cancer tissues, which is associated with poorly prognostic factors and poor prognosis, and it may be a therapeutic target in patients with gastric cancer.
ObjectiveTo explore the diagnostic value of ultrasound elastography (USE) combined with long non-coding RNA actin filament associated protein 1 anti-sense RNA 1 (AFAP1-AS1) mRNA in thyroid fine-needle aspiration (FNA) wash-out fluid for distinguishing benign from malignant thyroid nodules. MethodsThe patients with thyroid nodules who were treated in the Shenzhen Futian District Second People’s Hospital from January 2020 to June 2022 were collected. Before operation, the patients’ thyroid nodules were evaluated by the USE score and the AFAP1-AS1 mRNA in the thyroid FNA wash-out fluid was detected. The pathological result of the thyroid nodule after operation was as a gold standard for diagnosis of malignant thyroid nodules. The clinical diagnostic value of USE score combined with AFAP1-AS1 mRNA in the FNA wash-out fluid of the benign and malignant thyroid nodules were analyzed. ResultsA total of 174 thyroid nodules (124 patients) were detected in this study, of which 62 (45 patients) were histologically diagnosed as malignant. There was a statistical difference in the comparison of the composition ratio of USE score grading between the benign and malignant thyroid nodules (Z=8.82, P<0.001). The point of USE of the benign thyroid nodules was statistically lower than that of the malignant thyroid nodules [2.28±1.16 vs. 4.26±1.01, mean difference (MD) and 95% confidence interval (95%CI)=2.98 (2.76, 3.20), t=30.85, P<0.001]. The AFAP1-AS1 mRNA in the FNA wash-out fluid of the malignant thyroid nodules was statistically higher than that of the benign thyroid nodules [1.45±0.27 vs. 1.13±0.16, MD (95%CI)=1.45(1.39, 1.50), t=10.69, P<0.001]. Pearson correlation analysis showed that there was a positive correlation between the USE score of thyroid nodules and the expression of AFAP1-AS1 mRNA in the FNA wash-out fluid (r=0.58, P<0.001). The sensitivity and specificity of USE score in combination with expression of AFAP1-AS1 mRNA in the FNA wash-out fluid for diagnosing the malignant thyroid nodules by receiver operating characteristic (ROC) curve was 93.5% and 88.4% respectively. The area under the ROC curve (95%CI) was 0.91 (0.86, 0.96). Conclusion According to preliminary results of this study, USE score combined with AFAP1-AS1 mRNA in the thyroid FNA wash-out fluid is more sensitive and shows a potential diagnostic performance than USE score or AFAP1-AS1 mRNA detection alone for distinguishing benign from malignant thyroid nodules.
Objective To explore the expression characteristics of chaperone interacting protein (CHIP) in normal, scar and chronic ulcer tissues and its relationship with wound healing. Methods Twenty biopsies including scar tissues(n=8), chronic ulcer tissues(n=4) and normal tissues(n=8)were used in this study. The immunohistochemical staining (power visionTMtwo-step histostaining reagent) was used to explore the amount and expression characteristics of such protein.Results The positive expression of CHIP was observed in fibroblasts, endothelial cells and epidermal cells in dermis and epidermis. It was not seen ininflammatory cells. The expression amount of CHIP in scar tissues, chronic ulcer tissues and normal tissues was 89%, 83% and 17% respectively. Conclusion Although the function of CHIP is not fully understood at present, the fact that this protein is expressed only at the mitogenic cells indicates that it may be involved in mitogenic regulation during wound healing.
Vascular smooth muscle cells (VSMCs) phenotype switching plays an essential role in the pathogenesis of various vascular diseases. The present study aims to investigate the role of receptor-interacting protein kinases 1(RIPK1) in VSMCs phenotypic switching induced by Angiotensin Ⅱ(Ang Ⅱ). Expression of mRNA and protein of RIPK1, markers of VSMCs phenotypic switching and secretion, phosphorylation of the P65 subunit of NF-κB were measured by real-time PCR and Western blot. Meanwhile, EdU incorporation assay and wound scratch assay were performed to determine the cell proliferation and migration respectively. At the same time, Necrostatin-1(Nec-1, an known RIPK1 inhibitor) and RIPK1-specific small interference RNA (siRNA) were used to inhibit the expression of RIPK1. The experimental data demonstrated that the mRNA and protein levels of RIPK1 and P65 phosphorylation were increased significantly in the process of VSMC phenotypic switching induced by Ang II. Moreover, the expression of RIPK1 and P65 phosphorylation were significantly down-regulated in VSMCs pretreated with Nec-1 or trans-fected with RIPK1-siRNA. Furthermore, the proliferation, secretion and migration of VSMCs were also markedly suppressed after inhibition of RIPK1 by Nec-1 or its specific siRNA. The results suggested that RIPK1 might be involved in VSMC phenotypic switching induced by Ang II, which was possibly via up-regulating the NF-κB signaling pathway.
Objective Through studying a diabetic patient accompanied with pancreatic cancer by means of evidence-based clinical practice, to find out the relationship between diabetes mellitus and cancer and whether the long-acting insulin glargine increases the risk of cancer or not, which is regarded as a disputable hot issue at present. Methods Such databases as The Cochrane Library (Issue 3, 2010), OVID-EBM Reviews (1991 to Sept. 2010), MEDLINE (1950 to Sept. 2010) and CNKI (2000 to Sept. 2010) were retrieved to collect high quality clinical evidence, and the best therapy was formulated in accordance with the willingness of patients themselves. Results Eight randomized controlled trials (RCTs), four meta-analyses and one RCT meta-analysis were included. The evidence indicated that: a) Diabetes mellitus was kind of related to the occurrence of malignancies; b) There was no evidence at present showing the relationship between long-acting insulin glargine and cancer; c) Strictly controlling of blood sugar did not increase the risk of tumorigenesis, but hyperglycemia causing cancer was proofless; and d) Whether the diabetic patient with cancer should stop taking long-acting insulin glargine or not should require suggestions from specialists rather than patients themselves. Conclusion No evidence at present shows that tumorigenesis is related to diabetes mellitus, long-acting insulin glargine and strict controlling of blood sugar. It is necessary to require more evidence to decide whether the therapy should be adjusted or not for the diabetic patient with cancer who is in the process of glargine therapy.
Objective To explore the effect of cyclopamine （Cyc） which is the inhibitor of the Hedgehog signaling pathway on portal venous pressure of normal and liver cirrhosis rats, and it’s possible mechanisms. Moreover, to provide the experimental basis of drug efficacy and clinical treatment. Methods Thirty two healthy male SD rats were randomly average divided into four groups：normal control group, normal treatment group, liver cirrhosis control group, and liver cirrhosis treatment group. The liver cirrhosis models of rat were established by using the thioacetamide （TAA） method, which made 0.03% of TAA as the initial water concentration, and then the concentration of TAA in drinking water was adjusted according to the changes of the weekly body weight of rats lasting for twelve weeks. In thirteenth week, intraperitoneal injection of corn oil （0.1 ml/100 g body weight, 1 time/d） were performed lasting for a week in rats of the normal control group and liver cirrhosis control group; intraperitoneal injection of Cyc 〔1 mg （0.1 ml）/100 g body weight, 1 time/d〕were performed lasting for a week in rats of the normal treatment group and liver cirrhosis treatment group. In fourteenth week, the liver function, portal venous pressure （PVP）, and the ration of liver or spleen weight to body weight were detected, the expressions of α-smooth muscle actin （α-SMA） and typeⅠcollagen α1 （Col1α1） of hepatic stellate cell were detected by using immunohistochemistry. Results PVP were （10.7±0.9） and （12.3±1.3） cm H2O （1 cm H2O=0.098 kPa） in normal control group and normal treatment group, respectivly, the latter was higher than the former （t=-2.918，P=0.011）. PVP were （21.8±0.7） and （14.3±1.4） cm H2O in liver cirrhosis control group and liver cirrhosis treatment group, respectivly, the latter was lower than the former（t=13.602，P=0.000）. The expressions of α-SMA and Col1α1 in liver cirrhosis treatment group was lower than the liver cirrhosis control group. There were no significant difference of the liver function and ration of liver or spleen weight to body weight between the treatment group and the control group （P＞0.05）. Conclusion Cyclopamine could signally reduce the PVP of liver cirrhosis rats through reducing the expressions of α-SMA and Col1α1.
ObjectiveTo detect the expression of contactin-1 (CNTN1) gene in the breast cancer tissues and explore the relation between the expression and prognosis of patients with breast cancer. MethodsThe clinicopathologic data of 35 patients with breast cancer who met the inclusion criteria from January to June in 2015 in the Shaanxi Provincial Cancer Hospital were retrospectively collected. The Western blot and immunohistochemistry methods were used to detect the CNTN1 protein expressions in the 35 breast cancer tissues and their corresponding tissues adjacent to cancer and the reverse transcriptase-PCR method was used to detect the CNTN1 mRNA expression. The relation between the CNTN1 protein expression and 5-year overall survival (OS) was analyzed. ResultsThe positive rate and expression level of CNTN1 protein in the breast cancer tissues were higher than those in the tissues adjacent to cancer [65.7% (23/35) vs. 45.7% (16/35), χ2=8.235, P=0.003; 0.825±0.221 vs. 0.412±0.117, t=12.556, P<0.001], and the expression level of CNTN1 mRNA in the breast cancer tissue was also higher than that in the corresponding tissues adjacent to cancer (0.763±0.218 vs. 0.353±0.135, t=11.162, P<0.001). The positive rates of CNTN1 protein were higher in the patients with larger tumor diameter (>2 cm), lower differentiation, later TNM stage (stage Ⅲ+Ⅳ) and axillary lymph node metastasis (P<0.05). The median OS of 35 patients with breast cancer was 47.3 months, which was 36.2 months and 52.5 months in the patients with high CNTN1 expression and low CNTN1 expression (median expression of CNTN1 protein was 0.785 as critical value) respectively (χ2=4.052, P=0.049). ConclusionPreliminary results of this study suggest that overexpression of CNTN1 gene might play an important role in occurrence and progression of primary breast cancer and is related to prognosis of survival.
ObjectiveTo improve the knowledge of pulmonary actinomycosis.MethodsThree cases of pulmonary actinomycosis in this hospital and 65 cases reported in China were analyzed retrospectively.ResultsAmong the 68 patients 49 were male and 19 were female aged 6 to 77 years old. The most common clinical manifestations were cough, sputum and fever. Inflammatory indicators was slightly elevated. The most common site was on the right upper lung. The typical CT manifestations were the low-density liquefaction necrotic zone in the center of the mass with vacuoles of different sizes, namely, "air-space consolidation". Positron emission computed tomography showed a mild metabolic increase in lesions. The 68 patients were confirmed by surgery, CT guided percutaneous lung puncture or bronchoscopic biopsy. The average time of the diagnosis was 10 months while the longest time was 6.4 years. The rate of first diagnosis was 5.9%. Forty-one cases were treated with antibiotics alone and 12 cases were treated with simple operation, the rest were treated by antibiotics combined with surgical treatment. The cure rate was 88.7%. Although active treatment was conducted 3 patients in this hospital were not cured.ConclusionsThe clinical features of pulmonary actinomycosis are atypical and the misdiagnosis rate is high. When pulmonary actinomycosis is suspected, it should be fully communicated with the microbiologist to ensure the cultivation in anaerobic environment and extension of the incubation cycle. Tissue culture and pathological biopsy should be actively performed. Treatment depends on antibiotics or surgery with good prognosis, but for some cases the prognosis is not optimistic.
Objective To investigate the expression of high mobility group protein-B1( HMGB1)and α-smooth muscle actin( α-SMA) in Bleomycin induced pulmonary fibrosis in mice. Methods Twenty C57BL/ 6 male mice were randomly divided into a Bleomycin group and a control group. The Bleomycin group was treated with Bleomycin( 3 mg/kg) by endotracheally injection to induce pulmonary fibrosis. The control group were treated with normal saline( NS) . Then they were sacrificed by abdominal aortic bleeding 10 days after the injection. The right lung was stained with hematoxylin-eosin and Masson trichrome respectively for pathological examination. Immunohistochemistry and RT-PCR were performed to identify the protein and mRNA levels of α-SMA and HMGB1 respectively. Results The mRNA( 0. 89 ±0. 12, 0. 61 ±0. 08) and protein( 13. 66 ±1. 01, 13. 12 ±1. 33) expressions of α-SMA and HMGB1 in the Bleomycin group were all significantly higher than those of the control group( mRNA: 0. 60 ±0. 07, 0. 15 ±0. 02; protein: 8. 18 ±1. 33,7. 92 ±1. 10; all P lt; 0. 01) . Conclusions The expressions of HMGB1 and α-SMA are increased in Bleomycin induced pulmonary fibrosis. HMGB1 participates in the pathological process of pulmonary fibrosis probably by activation of the α-SMA expression.
ObjectiveTo investigate the mechanism of G protein coupled receptor kinase interacting protein 1 (GIT1) affecting angiogenesis by comparing the differentiation of bone marrow mesenchymal stem cells (BMSCs) differentiated into endothelial cells between GIT1 wild type mice and GIT1 gene knockout mice.MethodsMale and female GIT1 heterozygous mice were paired breeding, and the genotypic identification of newborn mice were detected by PCR. The 2nd generation BMSCs isolated from GIT1 wild type mice or GIT1 gene knockout mice were divided into 4 groups, including wild type control group (group A), wild type experimental group (group A1), GIT1 knockout control group (group B), and GIT1 knockout experimental group (group B1). The cells of groups A1 and B1 were cultured with the endothelial induction medium and the cells of groups A and B with normal cluture medium. The expressions of vascular endothelial growth factor receptor 2 (VEGFR-2), VEGFR-3, and phospho-VEGFR-2 (pVEGFR-2), and pVEGFR-3 proteins were detected by Western blot. The endothelial cell markers [von Willebrand factor (vWF), platelet-endothelial cell adhesion molecule 1 (PECAM-1), and vascular endothelial cadherin (VE-Cadherin)] were detected by flow cytometry. The 2nd generation BMSCs of GIT1 wild type mice were divided into 4 groups according to the different culture media: group Ⅰ, primary cell culture medium; group Ⅱ, cell culture medium containing SAR131675 (VEGFR-3 blocker); group Ⅲ, endothelial induction medium; group Ⅳ, endothelial induction medium containing SAR131675. The endothelial cell markers (vWF, PECAM-1, and VE-Cadherin) in 4 groups were also detected by flow cytometry.ResultsWestern blot results showed that there was no obviously difference in protein expressions of VEGFR-2 and pVEGFR-2 between groups; and the expressions of VEGFR-3 and pVEGFR-3 proteins in group A1 were obviously higher than those in groups A, B, and B1. The flow cytometry results showed that the expressions of vWF, PECAM-1, and VE-Cadherin were significantly higher in group A1 than in groups A, B, and B1 (P<0.05), and in group B1 than in groups A and B (P<0.05); but no significant difference was found between groups A and B (P>0.05). In the VEGFR-3 blocked experiment, the flow cytometry results showed that the expressions of vWF, PECAM-1, and VE-Cadherin were significantly higher in group Ⅲ than in groupsⅠ, Ⅱ, and Ⅳ, and in group Ⅳ than in groups Ⅰ and Ⅱ (P<0.05); but no significant difference was found between groups Ⅰ and Ⅱ (P>0.05).ConclusionGIT1 mediates BMSCs of mice differentiation into endothelial cells via VEGFR-3, thereby affecting the angiogenesis.