Objective Tissue engineered bone (TEB) lacks of an effective and feasible method of storage and transportation. To evaluate the activity of osteogenesis and capabil ity of ectopic osteogenesis for TEB after freeze-dried treatment in vitro and in vivo and to explore a new method of preserving and transporting TEB. Methods Human bone marrow mesenchymal stem cells (hBMSCs) and decalcified bone matrix (DBM) were harvested from bone marrow and bone tissue of the healthy donators. TEB was fabricated with the 3rd passage hBMSCs and DBM, and they were frozen and dried at extremely low temperatures after 3, 5, 7, 9, 12, and 15 days of culture in vitro to obtain freeze-dried tissue engineered bone (FTEB). TEB and FTEB were observed by gross view and scanning electron microscope (SEM). Western blot was used to detect the changes of relative osteogenic cytokines, including bone morphogenetic protein 2 (BMP-2), transforming growth factor β1 (TGF-β1), and insul in-l ike growth factor 1 (IGF-1) between TEB and FTEB. The ectopic osteogenesis was evaluated by the methods of X-ray, CT score, and HE staining after TEB and FTEB were transplanted into hypodermatic space in athymic mouse. Results SEM showed that the cells had normal shape in TEB, and secretion of extracellular matrix increased with culture time; in FTEB, seeding cells were killed by the freeze-dried process, and considerable extracellular matrix were formed in the pore of DBM scaffold. The osteogenic cytokines (BMP-2, TGF-β1, and IGF-1) in TEB were not decreased after freeze-dried procedure, showing no significant difference between TEB and FTEB (P gt; 0.05) except TGF-β1 15 days after culture (P lt; 0.05). The ectopic osteogenesis was observed in TEB and FTEB groups 8 and 12 weeks after transplantation, there was no significant difference in the calcified level of grafts between TEB and FTEB groups by the analysis of X-ray and CT score. On the contrary, there was no ectopic osteogenesis in group DBM 12 weeks after operation. HE staining showed that DBM scaffold degraded and disappeared 12 weeks after operation. Conclusion The osteogenic activity of TEB and FTEB is similar, which provides a new strategy to preserve and transport TEB.
ObjectiveTo systematically review the efficacy of exercise interventions on thoracic kyphosis in adults. MethodsThe PubMed, EMbase, Web of Science, CINAHL, Cochrane Library, CNKI, CBM, WanFang Data and VIP databases were electronically searched to collect randomized controlled trials (RCTs) related to the objective from inception to November 2022. Two reviewers independently screened literature, extracted data and assessed the risk of bias of the included studies. Meta-analysis was then performed by using RevMan 5.4 software. ResultsA total of 10 RCTs involving 482 patients were included. The results of meta-analysis showed that exercise interventions could reduce kyphosis angle (MD=−5.27, 95%CI −8.37 to −2.17, P<0.01) and improve quality of life (SMD=0.78, 95%CI 0.52 to 1.04, P<0.01) in thoracic kyphosis in adults, but there were no significant differences between the two groups in pain and physical function. ConclusionCurrent evidence suggests that exercise interventions can reduce thoracic kyphosis angle and improve quality of life in adults, but the effects on pain and physical function are unclear. Due to the limited quality and quantity of the included studies, more high quality studies are needed to verify the above conclusion.
Objective To evaluate the effects of knowledge about malaria after the implementation of Global Fund Malaria Project in Chongqing for future prevention and treatment of malaria. Methods Four counties were selected from the counties of Global Fund Malaria Project of Chongqing. Three towns were randomly selected in each selected county; one primary school and one high school were investigated in each selected town; one class was randomly selected in each selected school; and 30 students in each selected class were randomly selected. Then, three villages were randomly selected in each selected county; one group of villagers were randomly selected in each selected village; and 200 local residents were randomly investigated in each selected villager group. Before and after implementation of Global Fund Malaria Project, their awareness of malaria was investigated using a questionnaire. Results After implementation of the project, the pupils’ average awareness rate of knowledge about malaria rose from 58.94% to 89.96% (χ2=179.48, Plt;0.01) with average increase of 31.02%; the middle school students’ average awareness rate of rose from 52.83% to 86.06% (χ2=196.64, Plt;0.01) with average increase of 33.23%; the local residents’ average awareness rate of knowledge about malaria rose from 56.74% to 83.89% (χ2=1 281.70, Plt;0.01) with average increase of 27.15%. The accuracies of all respondents after project implement was higher than before. After the implementation, the accuracies of malaria transmission route, main clinical symptoms, and seeing doctor while malaria occurring were all above 80%, but the accuracy of the index of best preventive methods was less than 50%. Conclusion The implementation of Global Fund Malaria Project in Chongqing increase the awareness rate of malaria knowledge but the relevant education should be strengthened, so as to promote general population’s awareness of malaria prevention.
Objective The combined appl ication of green fluorescent protein (GFP) and confocal laser scanning microscope three-dimensional reconstruction (CLSM-3DR) were used to monitor the construction and in vivo transplantation of tissue engineered bone (TEB), to provide for technology in selection of scaffolds and three-dimensional constructional methods. Methods After bone marrow mesenchymal stem cells (BMSCs) were isolated from a 2-year-old green goat by a combination method of density gradient centrifugation and adherent culture, and the expressions of CD29, CD60L, CD45, and CD44 in BMSCs were detected by flow cytometry. Plasmid of pLEGFP-N1 was ampl ified, digested by enzymes (Hind III, BamH I, Sal I, and Bgl II), and identified. Transfection of pLEGFP-N1 into PT67 cells was performed under the help of l iposome. Positive PT67 cells were picked out with G418, and prol iferated for harvesting virus. Based on the titre of virus, after BMSCs were infected by virus containing pLEGFP-N1, GFP positive BMSCs were collected and prol iferated for seeding cells. TEB was fabricated by GFP positive BMSCs and decalcified bone matrix (DBM) and observed by CLSM-3DR for the evaluation of the distribution and prol iferation of seeding cells. After TEB was transplanted in the defect of goat femur, CLSM was used for observing the survival and distribution of GFP positive cells in the grafts. Results The isolated cells were fibroblast-l ike morphous, with the positive expression of CD29 and CD44, and negative expression of CD60L and CD45. The digested production of pLEGFP-N1 was collected for ionophoresis, whose results showed the correct fragment length (6 900 bp). The virus of pLEGFP-N1 was harvested by transfection of pLEGFP-N1 into PT67 cells and used for further infection to obtain GFP positive BMSCs. The prol iferated GFP positive BMSCs and DBM were used for fabrication of TEB. The distribution, prol iferation, and migration of BMSCs in TEB were observed by CLSM-3DR. GFP positive cells also were observed in images of TEB graft in goat femur 28 days after transplantation. Conclusion The BMSCs labeled by GFP in three-dimensional scaffold in vivo were monitored well by CLSM-3DR. It suggests a wide use potency in monitoring of three-dimensional cultured TEB.
目的 研究组织工程骨结合带锁髓内钉修复成年山羊大段负重骨缺损的可行性，探索更可行的技术路径。 方法 将24只成年山羊，通过骨髓穿刺法获取山羊骨髓间充质干细胞（BMSC），将体外扩增及成骨定向诱导的第2代BMSC与同种异体脱钙骨基质（DBM）通过双相接种法构建组织工程骨。24只成年山羊，以带锁髓内钉构建股骨中段3 cm骨缺损模型。随机分为3组，每组8只。实验组以组织工程骨修复骨缺损，对照组单独使用DBM和空白组旷置。术后1、12、24周行X线片观察及评分，12、24周每组各处死4只动物行组织学观察和生物力学检测。 结果 标本大体观察示实验组和对照组术后12周骨缺损部位被骨痂连接，髓腔贯通，24周全部愈合；实验组24周恢复正常解剖形态，对照组外形仍然粗糙、不规则；空白组术后12周及24周缺损部位均为纤维组织充填。术后1周各组X线评分无明显差异（P＞0.05），实验组术后12周及24周X线评分均优于对照组和空白组，对照组优于空白组，各组24周X线评分均高于12周时，差异均有统计学意义（P＜0.05）。实验组术后12、24周的最大抗扭强度分别达正常侧的47.07% ± 5.05%和83.73% ± 2.33%，显著高于对照组和空白组（P＜0.05）；空白组2个时间点最大抗扭强度均不超过正常的15%，与骨不连时的纤维连接相符。组织学检查示术后12周实验组和对照组骨缺损区DBM支架材料基本被吸收，有典型的同心圆排列的哈弗系统形成，周围偶见淋巴细胞；术后24周，实验组和对照组股骨缺损均被修复，但实验组较对照组的新骨更多、骨塑形更好；空白组术后24周骨缺损区中央仍为纤维组织填充。 结论 组织工程骨结合带锁髓内钉能够更有效修复成年山羊负重骨大段骨缺损，满足负重骨的生物力学要求。
Objective To learn and evaluate the effect of health education of schistosomiasis in the Three Gorges Reservoir Area, so as to provide scientific basis for establishing the comprehensive prevention and control model in potential epidemic area of schistosomiasis. Methods Through adopting the multi-stage stratified cluster sampling method, the questionnaires were distributed to residents selected randomly from the demonstration, inside and outside control areas. The survey data were input with EpiData 3.0 software by two reviewers, and the differences among groups were analyzed by using SPSS 18.0 software to further evaluate the effect of health education of schistosomiasis. Results All the questionnaires distributed to 1 420 residents were retrieved (100%). The results of analysis showed that for the residents in the demonstration area, the average score of being aware of schistosomiasis prevention was (7.51±2.89), which was markedly higher than those in the inside (2.52±2.97) and the outside (3.13±3.51) control areas, with significant differences (Plt;0.05); and the passing rates were 78.15%, which was also obviously higher than those in the inside (17.26%) and the outside control areas (32.16%), with significant differences (Plt;0.05). Conclusion The health education of schistosomiasis can improve residents’ knowledge about schistosomiasis prevention, and it is very important to prevent and control the potential prevalence of schistosomiasis in the Three Gorges Area.
Objective To learn the status and compare the effect of improving drinking water and lavatories in potential endemic areas of schistosomiasis in the Three Gorges Reservoir Area. Methods A cluster random sampling method was adopted and a questionnaire survey was conducted to four selected villages and towns before and after organized and large scale drinking water and lavatories improvement in the potential endemic areas of schistosomiasis in the Three Gorges Reservoir Area. Results A total of 807 households in four demonstrate villages and towns were investigated for the baseline survey, while 856 households in the same four villages and towns were investigated for the terminal survey. The popularity rate of using tap water from waterworks went up from 33.58% to 66.12% (P=0.000); the summation of popularity rate of using tap water from both waterworks and water tower made by local residents went up from 76.33% to 96.38% (P=0.000); and the popularity rate of using decontaminated lavatories went up from 53.68% to 78.85% (P=0.000). According to Chi-square test, there were significant differences compared with the popularity rate before improvement (Plt;0.05). Conclusion The condition of intervention villages and towns gets greatly improved through the organized and large scale project of improving water quality and lavatories decontamination, and the potential danger of schistosomiasis is obviously reduced in the Three Gorges Reservoir Area.
ObjectiveBased on the cell-extracellular matrix adhesion theory in selective cell retention (SCR) technology, demineralized bone matrix (DBM) modified by simplified polypeptide surface was designed to promote both bone regeneration and angiogenesis.MethodsFunctional peptide of α4 chains of laminin protein (LNα4), cyclic RGDfK (cRGD), and collagen-binding domain (CBD) peptides were selected. CBD-LNα4-cRGD peptide was synthesized in solid phase and modified on DBM to construct DBM/CBD-LNα4-cRGD scaffold (DBM/LN). Firstly, scanning electron microscope and laser scanning confocal microscope were used to examine the characteristics and stability of the modified scaffold. Then, the adhesion, proliferation, and tube formation properties of CBD-LNα4-cRGD peptide on endothelial progenitor cells (EPCs) were detected, respectively. Western blot method was used to verify the molecular mechanism affecting EPCs. Finally, 24 10-week-old male C57 mice were used to establish a 2-mm-length defect of femoral bone model. DBM/LN and DBM scaffolds after SCR treatment were used to repair bone defects in DBM/LN group (n=12) and DBM group (n=12), respectively. At 8 weeks after operation, the angiogenesis and bone regeneration ability of DBM/LN scaffolds were evaluated by X-ray film, Micro-CT, angiography, histology, and immunofluorescence staining [CD31, endomucin (Emcn), Ki67].ResultsMaterial related tests showed that the surface of DBM/LN scaffold was rougher than DBM scaffold, but the pore diameter did not change significantly (t=0.218, P=0.835). After SCR treatment, DBM/LN scaffold was still stable and effective. Compared with DBM scaffold, DBM/LN scaffold could adhere to more EPCs after the surface modification of CBD-LNα4-cRGD (P<0.05), and the proliferation rate and tube formation ability increased. Western blot analysis showed that the relative expressions of VEGF, phosphorylated FAK (p-FAK), and phosphorylated ERK1/2 (p-ERK1/2) proteins were higher in DBM/LN than in DBM (P<0.05). In the femoral bone defect model of mice, it was found that mice implanted with DBM/LN scaffold had stronger angiogenesis and bone regeneration capacity (P<0.05), and the number of CD31hiEmcnhi cells increased significantly (P<0.05).ConclusionDBM/LN scaffold can promote the adhesion of EPCs. Importantly, it can significantly promote the generation of H-type vessels and realize the effective coupling between angiogenesis and bone regeneration in bone defect repair.