Effects of growth factors (EGF,PDGF-BB and TGF-beta 1) on cultured equine epithelial cells and keratocytes: implications for wound healing

Objective The physiologic mechanisms involving growth factors, including PDGF‐BB, EGF, and TGF‐β1, as potent mediators of fibroblasts and epithelial cells in corneal wound healing remain unknown. The goal of this study was to determine culture methods for equine epithelial cells and keratocytes and to investigate how exogenous growth factors influence proliferation of both cell types. Procedures Cell cultures were established from healthy corneas harvested from horses immediately following euthanasia and maintained using standard tissue culture protocols. To determine the effects of PDGF‐BB, EGF, TGF‐β1, keratocytes (1 × 10⁵/well) and epithelial cells (2 × 10⁵/well) were each cultured in 12 well plates and exposed separately to the growth factors. The cells were exposed to concentrations of EGF between 0 and 50 ng/mL; PDGF‐BB between 0 and 75 ng/mL; and TGF‐β1 between 0 and 10 ng/mL. Cell proliferation was measured using ³H‐thymidine assay and differences in growth determined using anova and Tukey's HSD test (P < 0.05). Results Epithelial cell and keratocyte cultures were successfully established. EGF maximally stimulated keratocyte and epithelial cells at 25 ng/mL and 5 ng/mL, respectively. PDGF‐BB maximally stimulated keratocytes and epithelial cells at 50 ng/mL and 5 ng/mL, respectively. TGF‐β1 inhibited keratocytes at 5 ng/mL and 10 ng/mL, and epithelial cells at 1 ng/mL and 2 ng/mL. Conclusions Methods were established to maintain epithelial cells and keratocytes in vitro. PDGF‐BB and EGF stimulate, while TGF‐β1 inhibits the proliferation of epithelial cells and keratocytes. These growth factors may play a role in maintenance and repair of the equine cornea.     

社会转型时期加强中国共产党政党文化建设的思考

政党文化是指政党在长期的政治实践中形成和发展起来的,为全体党员普遍认同和共同遵循的政党意识和观念。作为一个自成体系的政治组织,中国共产党有自己的政党文化系统。社会转型是当前中国共产党政党文化建设的宏观生态。社会转型往往会出现认同、信仰、整合等危机。因此,社会转型时期,中国共产党合乎逻辑的选择是大力加强自身文化建设。     

影响螺旋藻生长因素的研究进展

螺旋藻具有丰富和全面的营养,同时具有重要的医疗保健价值,有着广阔的应用前景.笔者主要论述了不同的螺旋藻培养条件对其生长和产物的影响.     

发酵床垫料饲养荷斯坦牛试验报告

选择饲养密度、品种一致的荷斯坦牛,分为试验组和对照组,对照组砖面平养,试验组采用发酵床垫料,饲养试验中对产量、环境卫生等指标进行测定。结果显示,试验组的产奶量、料奶比等指标明显好于对照组;试验组牛舍NH3浓度低于对照组,粪便降解充分,奶牛均匀躺卧在垫料上。说明建立的试验方法可行,为生物环保养殖技术应用于奶牛生产提供参考依据。     

以超滤技术浓缩猪细小病毒含毒细胞液的试验

在研制猪细小病毒灭活疫苗时,应用了超滤浓缩技术,提高了合毒细胞培养液中的抗原含量。为了保证浓缩的流量,减少对滤膜的污染,对含毒细胞培养液作了预处理一离心及微滤,以便除去细胞培养液中的细胞碎片、蛋白质等大分子物质及固形微粒。超滤时选用50000Da滤膜,在20psig压力下,经过5倍浓缩的PPV细胞培养液,血凝滴度提高2个滴度以上。病毒含量测定表明,TCID50／0．2mL由107左右升至109以上,以其配制疫苗,能显著地提高疫苗的免疫原性。超滤技术具有易于操作、高效、分离精度高、没有二次污染等优点,可以根据需要选择不同的浓缩浓度,对保证疫苗的质量具有重要作用。     

Effect of dipeptide on intestinal peptide transporter 1 gene expression: An evaluation using primary cultured chicken intestinal epithelial cells

Peptide transporter 1 (PepT1) is a transporter responsible for absorbing dipeptide and tripeptide in enterocytes and is upregulated by dipeptide in mammals. It has not been certain whether intestinal PepT1 expression is responsive to dipeptides in chickens because of the lack of in vitro study using the cultured enterocytes. This study established a primary culture model of chicken intestinal epithelial cells (IECs) in two-dimensional monolayer culture using collagen gel by which the response of chicken PepT1 gene expression to dipeptide stimuli was evaluated. The cultured chicken IECs showed the epithelial-like morphology attached in a patch-manner and exhibited positive expression of cytokeratin and epithelial cadherin, specific marker proteins of epithelial cells. Moreover, the chicken IECs exhibited the gene expression of intestinal cell type-specific marker, villin1, mucin 2, and chromogranin A, suggesting that the cultured IECs were composed of enterocytes as well as goblet and enteroendocrine cells. PepT1 gene expression was significantly upregulated by synthetic dipeptide, glycyl-l-glutamine, in the cultured IECs. From the results, we herein suggested that dipeptide is a factor upregulating PepT1 gene expression in chicken IECs.     

A filter-assisted culture method for isolation of Treponema spp. from bovine digital dermatitis and their identification by MALDI-TOF MS

Digital dermatitis (DD) is a major infectious foot disease of cattle worldwide. Some DD stages are associated with lameness, and the disease has significant economic and animal welfare consequences. The pathogenesis of the disease is not yet fully understood, but Treponema spp. have been associated consistently with clinical cases. Isolation of these fastidious bacteria is difficult and cumbersome. We describe an improved method enabling the culturing of the 3 Treponema spp. (T. pedis, T. phagedenis, and T. medium) from bovine foot specimens derived from DD lesions, using a combination of membrane filtering and subsequent growth on selective agar media. The entire procedure from sampling to verification of individual Treponema spp. takes up to 24 d. In addition, we established a MALDI-TOF MS–based identification method to be applied for confirmation of the different Treponema spp. This scheme provides an unambiguous, simple, and straightforward identification procedure for DD-associated Treponema spp.     

鹿茸干细胞体外培养技术的研究

利用消化酶消化法,用DMEM培养基对鹿茸干细胞进行了体外培养,分别从原代培养、传代、冻存、以及复苏等几个环节进行了试验。摸索鹿茸干细胞合适的体外培养条件,为鹿茸的各项相关研究打基础。结果表明,用透明质酸酶和胶原酶对组织样进行消化,然后用10%DMEM培养基进行培养细胞生长效果较好。适宜培养条件是37℃、5%CO2最大饱和湿度。     

利用IVM/IVF技术生产试管犊牛的研究

本研究利用卵母细胞体外成熟(IVM)、体外受精(IVF)技术,体外生产牛胚胎,以获得试管牛.结果表明:卵母细胞体外培养22～24h后,其成熟率为85％;体外受精率为83％.体外受精卵分别在颗粒细胞单层(GCM)和输卵管细胞单层(OCM)上培养,其胚胎最后产量(以授精时卵母细胞的数目为基数)分别为19％和29％,差异极显著(P<0.01).若体外受精卵先在GCM中培养72h后,再将已发生卵裂的(>4细胞)胚胎移到OCM中培养,其胚胎最后产量为35％.由此表明,早期胚胎在其发育过程中至少存在着3个发育相期,即1～8细胞、8～16细胞和16细胞～囊胚3个阶段,各阶段需要不同的培养环境.IVM/IVF胚胎移植到受体黄牛体内后,其足月时的妊娠率为15％.第一头试管犊牛已于1993年4月18日凌晨产出.     

Salmonella enterica infections in Spanish swine fattening units

The present study is the first conducted in Spain to estimate the bacteriological herd prevalence of Salmonella enterica in fattening units and to describe the Salmonella serovar diversity on these farms using a sample representative of the entire swine population. For this purpose, 10 faecal samples were collected from 10 different pens containing pigs close to market weight in a total of 232 fattening units. Total sample size was proportionally distributed according to the fattener census in each of the regions of the country and all the samples were examined by culture of 25 g of faecal material. One hundred (43.1%) farms had at least one Salmonella-positive sample (95% CI: 37-49.1%). Salmonella enterica was detected in 290 (12.5%) pooled faecal floor samples (95% CI: 11.2-13.8%). The apparent herd prevalence of salmonellosis was similar among multi-site, finishing and farrow to finish farms. Overall, 24 different serovars were identified, with S. Typhimurium, S. Rissen and S. Derby being the most common both at herd and sample level. Results of phage typing were available for the 91 isolates of S. Typhimurium. A total number of 10 different phage types were identified, with DT 193 being the most frequent. Phage types DT 104, DT 104b and DT U302, which have been associated with several multi-resistant patterns, accounted for 23% and 29% of the Typhimurium total isolates or Typhimurium infected farms respectively.