Objective To investigate the effects of simvastatin on the collagen synthesis of rat pulmonary arterial smooth muscle cells ( PASMCs ) induced by hypoxia. Methods Under hypoxic condition, rat PASMCs were cultured with different concentrations of simvastatin. Collagen synthesis of PASMCs with or without simvastatin were measured by 3H-proline incorporation assay. The mRNA expression of TGF-β1 and the contents of super oxide dismrtase ( SOD) ,malondialdehyde ( MDA) in mediumwere also measured. Results The incorporation data of 3H-TdR in the hypoxia group was significantly increased as compared with that in the control group ( P lt;0. 01) , and simvastatin significantly reduced the incorporation data of 3H-TdR induced by hypoxia. The expression of TGF-β1 mRNA in the hypoxia group was significantly increased as compared with that in the control group ( P lt; 0. 01 ) , and simvastatin could significantly inhibited hypoxia-induced expression of TGF-β1 mRNA in a dose-dependent manner. Compared with the hypoxia group, the expression of TGF-β1 mRNA decreased by 55% in simvastatin( 10 - 6mol /L) group ( P lt; 0. 01) , and by 70% ( P lt; 0. 01) in simvastatin ( 10 - 5mol /L) group. Compared with the control group, the activity of SOD was reduced and the contents of MDA were increased significantly in the hypoxia group. Simvastatin can increase the activity of SOD and reduced the content of MDA in a dose-dependent manner. Conclusions Simvastatin can decreases collagen synthesis of PASMCs. This effect might be explained that simvastatin can reduce lipid peroxide and expression of TGF-β1 mRNA.
在缺乏数据进行 Meta 分析的系统评价中，通常使用替代合成方法，但这些方法却很少被报道，而模糊的方法阐述可能会导致人们质疑系统评价结果的真实性。无 Meta 分析数据合成（SWiM）报告规范是用于指导采用了替代合成方法评估干预措施效应的系统评价进行清晰报告的规范。本文介绍了 SWiM 规范的研制过程及 9 个 SWiM 报告条目及其相应的解释与示例。
Objective Astragalus polysaccharide (APS) has promoting angiogenesis function. To explore the effects of APS collagen sponge on enhancing angiogenesis and collagen synthesis so as to provide evidence for the future tissue engineering appl ication as a kind of angiogenic scaffold. Methods APS collagen sponges were prepared by covalent binding with collagen polypeptides by using of crossl inking agents at the ratio of 1 ∶ 1 (W/W). Twenty 10-week-old SpragueDawley rats (10 males and 10 females, and weighing 200-250 g) were selected. Longitudinal incision was made at both sides of the back to form subcutaneous pockets. APS collagen sponges of 5 mm × 5 mm × 5 mm at size were implanted into the left pockets as the experimental group, collagen sponges without APS of the same size into the right pockets as the control group. The general conditions were observed after operation. At 3, 7, 14，and 21 days, 5 rats were sacrificed and the samples were harvested to count the number of microvessels, to measure the contents of the hydroxyprol ine (Hyp), and to detect the mRNA expressions of angiopoetin 1 (Ang1), matrix metalloproteinases 9 (MMP-9), and tissue inhibitors of metalloproteinases 1 (TIMP-1). Results All rats were al ive during experiment period. The number of microvessels increased gradually, and reached the peak at 14 days in 2 groups; the expermental group was significantly higher than the control group (P lt; 0.05). The contents of Hyp increased gradually in 2 groups, and the experimental group was significantly higher than the control group (P lt; 0.05). The mRNA expressions of Ang1 and MMP-9 in the experimental group were significantly higher than those in the control group at 3, 7, and 14 days (P lt; 0.05); the mRNA expression of TIMP-1 in the experimental group was significantly lower than that in the control group at 3 days and was significantly higher at 14 and 21 days (P lt; 0.05). Conclusion The APS collagen sponges can improve angiogenesis and collagen synthesis in wound heal ing by regulating the expressions of Ang1, MMP-9, and TIMP-1.
Objective To observe the mRNA and protein expression of wingless-type MMTV integration site family member 5a (Wnt5a), glycogen synthase kinase 3 (GSK3), and β-catenin, as well as the muscle fibers and adipose tissue presented in pathological staining in the gastrocnemius muscle of white rabbits with blunt gastrocnemius contusion injuries, and provide a basis for revealing the repair mechanism of the pressing and kneading method in treating skeletal muscle injury. Methods Forty-two healthy male and female New Zealand white rabbits were selected. They were randomly divided into blank group, model 3-day group, model 7-day group, model 14-day group, press-and-knead 3-day group, press-and-knead 7-day group, and press-and-knead 14-day group, by using a random number table method, with 6 rabbits in each group. Samples of the model groups and the press-and-knead groups were taken on the 4th, 8th and 15th days after operation. The mRNA and protein expression of Wnt5a, GSK3, and β-catenin were detected by quantitative polymerase chain reaction and Western blot; the muscle tissue myofibers and adipose tissue were observed by hematoxylin and eosin (HE) staining and oil red O staining. Results The HE staining results showed that significant fibrous tissue proliferation and inflammatory cell infiltration occurred in the model 7-day group; in the model 14-day group, some muscle fibers were degenerated, necrotic, and regenerated, accompanied by fibrous tissue proliferation, slight inflammatory cell infiltration, and slight calcification; in the press-and-knead groups, obvious muscle fiber degeneration, necrosis, and regeneration, and inflammatory cell infiltration were observed, accompanied by significant fibrous tissue proliferation. The oil red O staining results showed that adipocyte deposition was visible in the model groups, which was the heaviest in the model 7-day group; in the press-and-knead groups, muscle fibers and sequences were not significantly damaged, and a small amount of adipocyte infiltration was visible in the interstitial space. There were statistically significant differences in the mRNA expression and protein expression of Wnt5a, GSK3, and β-catenin in the gastrocnemius among groups (P<0.001). Conclusions The histopathological changes of gastrocnemius muscle injury recover gradually over time, and the pressing and kneading method stimulates the mRNA expression activities of Wnt5a, GSK3, and β-catenin, which may slow down the degradation of β-catenin protein by the scaffolding protein complex (of which GSK3 is an important component), so that the protein level of β-catenin is maintained in the stable range at all times. This leads to a reduction of fatty degeneration in the gastrocnemius muscle after the intervention of pressing and kneading method, and promotes the functional repair of the injured skeletal muscle.
Objective To review the current status and problems in the developing scaffolds for the myocardial tissue engineering appl ication. Methods The l iterature concerning the myocardial tissue engineering scaffold in recent years was reviewed extensively and summarized. Results As one of three elements for tissue engineering, a proper scafold is veryimportant for the prol iferation and differentiation of the seeding cells. The naturally derived and synthetic extracellular matrix (ECM) materials aim to closely resemble the in vivo microenvironment by acting as an active component of the developing tissue construct in myocardial tissue engineering. With the advent and continuous refinement of cell removal techniques, a new class of native ECM has emerged with some striking advantages. Conclusion Through using the principle of composite scaffold, computers and other high-technology nano-polymer technology, surface modification of traditional biological materials in myocardial tissue engineering are expected to provide ideal myocardial scaffolds.
ObjectiveTo study the expression of urokinase-type plasminogenactivator (uPA) and phosphorylation of glycogen synthase kinase-3β (P-GSK3β) in human colorectal adenocarcinoma and its significance. MethodsSeventy-eight samples of colorectal adenocarcinoma got during operation between January 2006 and December 2010 in Handan Central Hospital were chosen as the study subjects. The immunohistochemical SP method was used to detect uPA and P-GSK3β levels in the 78 cases of colorectal adenocarcinoma, 20 cases of normal colorectal mucosa and 30 cases of colorectal adenoma. ResultsThe positive expression rates of uPA and P-GSK3β in colorectal carcinoma were much higher than those in colorectal mucosa, colorectal polyps, and colorectal adenoma (P<0.05). The expressions of uPA and P-GSK3β were closely correlated with the differentiation, TNM and lymph nodes metastasis (P<0.05). ConclusionThe expression of uPA and P-GSK3β is closely related to the colorectal adenocarcinoma occurrence. Both of them are important biological markers in colorectal adenocarcinoma occurrence and development.
Objective To investigate the effects of NGF on the prol iferation, mitotic cycle, collagen synthesis and migration of human dermal fibroblasts (HDFs), and to explore the function of NGF on the wound heal ing. Methods The 3rd generation of HDFs were incubated with various concentrations of NGF (0, 25, 50, 100, 200 and 400 ng/mL), the cell prol iferation was measured with MTT assay. After treated with NGF at 0, 100 ng/mL, the cell cycle of HDFs was determined by flow cytometry (FCM). Hydroxyprol ine and real-time fluorescence quantitative PCR (FQ-PCR) were used to measure collagen synthesis at protein level and mRNA level respectively. The in vitro cell scratch wound model was set up to observe the effect of NGF (0, 50, 100 and 200 ng/mL) on the migration of HDFs after 24 hours of culture. Results Absorbance value of HDFs for different concentrations of NGF (0, 25, 50, 100, 200, and 400 ng/ mL) showed that NGF did not influence the prol iferation of HDFs (P gt; 0.05). When HDFs were treated with NGF at 0 and 100 ng/mL, the result of FCM analysis showed that percentage of HDFs in G0/G1, S, G2/M phases were not changed (P gt; 0.05). Compared with control group, the expression of Col I and Col III were not significantly different, measured by both hydroxyprol ine and FQ-PCR (P gt; 0.05). The rates of HDFs’ migration at various concentrations of NGF (0, 50, 100, 200 ng/ mL) were 52.12% ± 6.50%, 80.67% ± 8.51%, 66.33% ± 3.58%, and 61.19% ± 0.97%, respectively, indicating that NGF could significantly enhanced the migration of HDFs at 50 and 100 ng/mL (P lt; 0.05). Conclusion NGF does not influence prol iferation, mitotic cycle and collagen synthesis of HDFs, but significantly enhanced migration in an in vitro model of wounded fibroblasts.
ObjectiveTo compare the biological features of early and late endothelial progenitor cells (EPCs) by isolating and culturing early and late EPCs from the human peripheral blood so as to find some unique properties of EPCs and to propose a suitable strategy for EPCs identification. MethodsMononuclear cells were isolated from the human peripheral blood using density gradient centrifugation. Then, the cells were inoculated in human fibronectin-coated culture flasks and cultured in endothelial cell basal medium 2. After 4-7 days and 2-3 weeks culture, early and late EPCs were obtained respectively. The morphology, proliferation potential, surface markers, cytokine secretion, angiogenic ability, and nitric oxide (NO) release were compared between 2 types of EPCs. Meanwhile, the human aortic endothelial cells (HAECs) were used as positive control. ResultsThe morphology of early and late EPCs was different:early EPCs formed a cell cluster with a spindle shape after 4-7 days of culture, and late EPCs showed a cobblestone appearance. Late EPCs were characterized by high proliferation potential and were able to form capillary tubes on Matrigel, but early EPCs did not have this feature. Both types EPCs could ingest acetylated low density lipoprotein and combine with ulex europaeus Ⅰ. Flow cytometry analysis showed that early EPCs did not express CD34 and CD133, but expressed the CD14 and CD45 of the hematopoietic stem cell markers;however, late EPCs expressed CD31 and CD34 of the endothelial cell markers, but did not express CD14, CD45, and CD133. By RT-PCR analysis, the expressions of vascular endothelial growth receptor 2 and vascular endothelial cadherin in early EPCs were significantly lower than those in the late EPCs and HAECs (P<0.05), but no significant difference was found in the expression of von Willebrand factor and endothelial nitric oxide synthase (eNOS) between 2 type EPCs (P>0.05). The concentrations of vascular endothelial growth factor, granulocyte colony-stimulating factor, and interleukin 8 were significantly higher in the supernatant of early EPCs than late EPCs (P<0.05). Western blot assay indicated eNOS expressed in both types EPCs, while the expression of eNOS in late EPCs was significantly higher than early EPCs at 5 weeks (P<0.05). Both cell types could produce similar amount of NO (P>0.05). ConclusionThe expression of eNOS and the production of NO could be used as common biological features to identify EPCs, and the strategy of a combination of multiple methods for EPCs identification is more feasible.