Objective To investigate the expression of vascular endothelial growth factor-A （VEGF-A） and the phenotypic transition after the activation of fibroblasts by the supernatant of cultured tumor cells．Methods The growth tendency of fibroblasts was tested by the MTT assay．The expressions of alpha-smooth muscle actin （α-SMA） and VEGF-A mRNA were tested by RT-PCR．The expressions of α-SMA and VEGF-A protein were tested by immunohistochemistry and Western blot．Results The MTT assay indicated that the conditional medium which contained tumor cells supernatant could obviously promote the growth of the fibroblasts． RT-PCR and Western blot manifested that α-SMA expressed by the fibroblasts which cultured by normal medium reached its peak on day 5，then decreased to a low level on day 7．When the medium contained 2 ng/ml transforming growth factor-β1 （TGF-β1），the fibroblasts could steadily express more α-SMA．But the above two mediums could not make the fibroblasts express the VEGF-A. When using the conditional medium，the α-SMA peak advanced on the third day and maintained at a high level，so as the expression of the VEGF-A．Conclusions The results suggested that fibroblasts can be activated to be myofibroblasts when using the conditional medium．The best activation time of the fibroblasts is consistent with the time of the VEGF-A expression at the highest level by the activated fibroblasts．The fibroblasts which activated at the best time are expected to become a kind of cells which can be used for promoting revascularization．
ObjectivesTo systematically review the methods of pharmacoeconomic evaluation model for hepatitis C therapies and to identify shortcomings of the existing modeling research by comparing the model structure, hypothesis and methodological differences, and to provide suggestions for the construction of high-quality hepatitis C pharmacoeconomic evaluation models.MethodsPubMed, EMbase, The Cochrane Library, CNKI and WanFang Data databases were electronically searched to collect relevant literatures on the pharmacoeconomic evaluation models for hepatitis C therapies from August 2014 to August 2019. Two reviewers independently screened literature, extracted data, and evaluated the quality of the included studies. Then, the data related to the model structure, methods, and assumptions were compared and summarized.ResultsMost of the 46 studies that finally included used similar modeling methods. Ignoring different modeling elements would cause overestimation or underestimation of the value of hepatitis C therapies. Model structure of all studies were similar and key parameters were from the same source. Forty-five studies measured the cost of drugs and medical cost of health status. All studies used quality-adjusted life years as the outcome and reported incremental cost-effectiveness ratio. Thirty studies conducted one-way sensitivity analysis and probability sensitivity analysis.ConclusionsThe included studies share similar methodological designs and have high quality in general. However, there are some differences and deficiencies in research perspective, model types, model assumptions and model verification. Future pharmacoeconomic evaluation model of hepatitis C therapies should report the results of the whole society, establish dynamic model to consider the impact of transmission, make half-cycle correction for long periods, consider the recurrence after cure, model liver transplantation, and verify the model.
ObjectiveTo investigate the expression of tumor suppressor gene Tat interacting protein 30 (TIP30) gene in papillary thyroid carcinoma and it’s clinical significance in treatment of thyroid carcinoma. Methods Thirty cases of pathological specimens wax pieces of papillary thyroid carcinoma from 2003 to 2006 in our hospital were selected, in which there were 7 male, 23 female； and the age was from 15 to 70 years old, average 44.7 years. Six cases were nodular goiter with carcinomatous change in local area (papillary), 2 cases were thyroid capsular invasion. Distant lymph node metastasis and lesions surrounding the thyroid tissue were not confirmed by pathology. Every specimen was divided into tumor tissue and adjacent tissue (1-2 cm far away from tumor and non-cancerous tissue was confirmed by pathology). The expression of TIP30 in specimen was detected by immunohistochemical method with staining index and the average absorbance. ResultsTIP30 was expressed in the cell membrane and cytoplasm, which was showed as brown particles. ①Staining index: TIP30 in adjacent tissues was expressed highly with 21 (70.0%) positive cases (gt;2 points) and 9 (30.0%) negative cases (≤2 points), while its expression in cancer tissues was reduced or missed with 11 (36.7%) positive cases (gt;2 points) and 19 (63.3%) negative cases (≤2 points). There was a statistical difference between them (P＜0.05), and it was not related to age and gender of patients (Pgt;0.05). ②The average absorbance of TIP30 in cancer tissues was significantly lower than that in adjacent tissue (P＜0.05). ConclusionThe expression of TIP30 in papillary thyroid carcinoma is reduced or deleted, which can supply some theory support for its gene therapy.
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.
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 To investigate the modulating effect of transforming growth factor beta;2 (TGFbeta;2) and extracellular matrix (ECM) on the transdifferentiation of human fetal RPE (hfRPE) cells into myofibroblast-like cells , and to determine the mechanism of signal transduction. Methods hfRPE cells were cultured on ECM coated or uncoated petri dish with or witho ut TGFbeta;2 in the medium. The expression of alpha;-smooth muscle actin (alpha;-SMA) were detected by immunocytochemistry examination, flow cytometry and Western blotting via calphostin C, genistein, PD98059, and Wortmannin. Results After cultured on ECM coated petri dish with TGFbeta;2 in the medium,there were obvious morphological changes of hfRPE cells including cellular elongating and appearing of actin microfilaments. The results of flow cytometry and immunocytochemistry examination showed that expression of alpha;-SMA obviously increased after TGFbeta;2 was added in the medium in a dose-dependent manner. Compared with which of hfRPE cells cultured on the uncoated surface of culture plates, the total mean fluore scence intensity (TMFI) of hfRPE cells cultured on FN-coated surface increased (38.01plusmn;1.14)% when the stimulation concentration of TGFbeta;2 was 50ng/ml(Plt;0.05). Western blotting further confirmed the effects. The changes mentioned above could be inhibited mostly by protein kinase C (PKC) and calphostin C (10 nmol/L)(Plt;0.01). Conclusion TGFbeta;2 may induce the transdifferentiation of hfRPE cells into myofibroblast-like cells in a dose dependent manner, which could be intensified by FN. These mediated effects of TGFbeta;2 and ECM may act via the PKC signal transduction pathway. (Chin J Ocul Fundus Dis, 2006, 22: 328-332)
ObjectiveTo evaluate the role of triclosan-coated polyglactin 910 suture in reducing wound infections of emergency gastrointestinal surgeries. MethodsThis was a prospective, randomized, controlled, single center study. From May 2009 to August 2010, 412 patients underwent emergency gastrointestinal operations in our department, 198 of them were chose randomly as experimental group using triclosancoated polyglactin 910 suture for abdominal wall closure， 214 using traditional braiding suture were taken as control. The risk factors for wound healing were analyzed, and wound infection rate was compared between two groups. ResultsThere were no significant differences of gender, age, body mass index, combined diabetes, use of immunosuppressant, and glucocorticoid steroid, type of incision, intraoperative bleeding volume, and operation time between two groups (Pgt;0.05). Wound infection rate of experimental group 〔3.0% (6/198）〕 was significantly lower than that of control group 〔11.7% (25/214), Plt;0.001〕. Especially in subgroup of type Ⅲ incision and operative time more than 120 min, wound infection rate was significantly different between experimental group and control group 〔3.5%(5/141) versus 14.3%(22/154); 3.3%(2/60) versus 21.2%(11/52) respectively, Plt;0.001〕. ConclusionTriclosancoated polyglactin 910 suture can reduce wound infection rate of gastrointestinal emergency operations, especially with type Ⅲ incision and operation time ≥120 min.
Objective To investigate the effect of hydrostatic pressure and insulin-like growth factor 1 (IGF-1) on the expression of filamentous actin (F-actin) of temporomandibular joint disc cells in goats, and to analyze the F-actin changes of temporomandibular joint disc cells in vitro under hydrostatic pressure and IGF-1 stimulation. Methods The bilateral temporomandibular joint discs were harvested from 4 1-month-old goats, and temporomandibular joint disc cells were isolated with collagenase. Immunohistochemical staining for collagen type I and collagen type II was performed for identification. The cells at passages 2-3 were used; the experiment was divided into 4 groups according to different interventions: the cells were cultivated with complete medium in group A as control; the cells were intervened by hydrostatic pressure (0.2 MPa and 1 Hz for 3 hours) in group B, by complete medium containing IGF-1 (10 ng/mL) in group C, and by a combination of hydrostatic pressure (0.2 MPa and 1 Hz for 3 hours) and complete medium containing IGF-1 (10 ng/mL) in group D. The changes of F-actin at 24 and 72 hours after cultivation were observed by immunofluorescence staining. The cell fluorescence intensity was measured. Results The cultivated cells were identified to be temporomandibular joint disc cells by morphological observation and immunohistochemical staining. At 24 hours, fluorescence intensity of groups A and C was b and clear, with normal morphology of temporomandibular joint disc cells; F-actin arranged in disorder in group B, and F-actin was thinner with arrangement disorder in group D. At 72 hours, the F-actin arranged regularly in groups A and C; however, some F-actin became blurry with irregular arrangement, breakage, and pseudopodia in group B; and F-actin was thinner and ruptured formed in group D. With time passing, the fluorescence intensity of F-actin in groups A, B, and D had an increasing trend, showing significant differences between 24 hours and 72 hours (P lt; 0.05); but there was no significant difference between 24 and 72 hours in group C (t=0.284, P=0.781). At 24 hours, fluorescence intensity of F-actin was highest in group C and was lowest in group B, showing significant difference when compared with groups A and D (P lt; 0.05). At 72 hours, fluorescence intensity in groups B and D was significantly lower than that in groups A and C (P lt; 0.05), but there was no significant difference between groups B and D, and between groups A and C (P gt; 0.05). Conclusion Hydrostatic pressure may cause the F-actin breakage and rearrangement of temporomandibular joint disc cells, and IGF-1 can up-regulate the F-actin expression. Such effects may be correlated with the biological behavior of the temporomandibular joint disc cells.
ObjectiveTo investigate the inhibitory effect of short hairpin RNA (shRNA) mediated contactin-1 (CNTN1) gene silencing on growth of human breast cancer cell line MDA-MB-468 transplanted tumors in nude mice.MethodsEighteen nude mice (4-week-old male BALB/c) were randomly equally divided into three groups: blank control group, empty vector group, and silencing group. The MDA-MB-468 cells (blank control group), MDA-MB-468 cells transfected by nonsense shRNA (empty vector group), and MDA-MB-468 cells transfected by shRNA (silencing group) were collected in the logarithmic growth period, respectively. The subcutaneous tumor models of nude mice were prepared by the subcutaneous injection of the different group cells. The tumor growth was observed and the expressions of CNTN1 and Ki-67 proteins in the transplanted tumor were detected by the immunohistochemistry.ResultsThe xenograft models of human breast cancer cells were established successfully. The tumor growth in the silencing group was significantly slower than that of the other two groups at every 3 d point (P<0.05). The tumor volume and the tumor weight in the silencing group were significantly smaller or slighter than those of the other two groups at day 18 (P<0.05). The positive rates of CNTN1 and Ki-67 protein expressions in the tumor tissues of the silencing group were lower than those of the other two groups (P<0.05), respectively.ConclusionSilencing expression of CNTN1 gene might inhibit growth of breast cancer cell line MDA-MB-468 transplanted tumors in mude mice.
Objective To explore whether epithelial to mesenchymal transition ( EMT) occurs in bleomycin( BLM) induced pulmonary fibrosis, and the involvement of bronchial epithelial cells( BECs) in the EMT. Methods BLM-induced peribronchial fibrosis in an α-smooth muscle actin-Cre transgenic mouse( α-SMACre /R26R) was examined by pulmonary βgal staining and α-SMA immunofluorescence staining. Results BLMtreated mice showed significantly enhanced βgal staining in subepithelial areas in bronchi, terminal bronchioles and walls of pulmonary vessels. Some alveolar epithelial cells( AECs) in certain peribronchial areas or even a small subset of BECs were also positively stained, as confirmed by α-SMA immunostaining. Conclusions EMT occurs in BLM-induced peribronchial fibrosis mice. BECs, like AECs, have the capacity to undergo EMT and to contribute to mesenchymal expansion in pulmonary fibrosis.