骨髓间充质干细胞移植在卵巢早衰小鼠卵巢功能重建中应用

探讨骨髓间充质干细胞(BMSCs)移植在卵巢早衰小鼠卵巢功能重建中的应用价值。通过一次性腹腔注射环磷酰胺和白消安建立小鼠卵巢早衰(POF)模型,并随机分为建模组、BMSCs移植组和空白对照组。移植组于建模14 d经尾静脉注射绿色荧光蛋白(GFP)转基因鼠BMSCs悬液(5×10~7个·mL~(-1)),每4 d重复1次,共5次;建模组和空白对照组注射等量生理盐水。观察建模后及BMSCs移植后小鼠的一般情况、血清FSH和E2水平、卵巢组织学变化以及GFP荧光信号在卵巢内的分布。结果发现,建模组小鼠食欲明显减退,活动减少,与空白对照组相比体重显著减轻(P0.05),FSH水平显著增高(P0.05),E2水平显著降低(P0.05),卵巢体积减小,间质纤维化严重,始基卵泡、生长卵泡和成熟卵泡数均显著减少(P0.05)。BMSCs移植后,移植组小鼠体重增加,食欲增强,活动增多;28 d后FSH浓度显著低于建模组(P0.05),E2浓度、始基卵泡、生长卵泡数及成熟卵泡数均显著高于建模组(P0.05),但各项指标仍与空白对照组间存在显著性差异;荧光显微镜下可见绿色荧光信号散在分布于卵泡周围。此结果提示,BMSCs可在小鼠损伤卵巢组织中定位存活,对卵巢组织结构及内分泌功能有修复作用。     

国际人体肝移植研究的文献计量分析

采用文献计量学方法,对PubMed收录的人体肝移植论文的时间、国家、机构、期刊进行分析,了解国际人体肝移植研究的发展轨迹、开展人体肝移植的主要国家、机构。综合运用共词分析、聚类分析和可视化分析方法及工具,对肝移植预后与生存研究的论文进行主题分析,并对结果进行可视化展示,更加准确、直观地揭示影响人体肝移植预后生存的主要因素及其研究热点。     

羽叶薰衣草的组织培养

以羽叶薰衣草(Lavandula pinnata L.)茎段为外植体进行离体培养,探讨羽叶薰衣草外植体的消毒灭菌、增殖培养、生根培养、炼苗移栽等一系列技术措施。结果表明:用0.1%HgCl2处理羽叶薰衣草外植体6~8 min的效果较好;外植体在含0.8 mg/L 6-BA及0.1 mg/LNAA的MS培养基中培养,增殖效果较好,培养42 d后,平均增殖系数达11.3;较合适的试管苗生根培养基为含0.2 mg/L NAA的1/2 MS培养基,培养21 d后,生根率为98.2%,平均每株生根12.5条;炼苗后移栽成活率达95.0%以上。     

Urinary Markers in Healthy Young and Aged Dogs and Dogs with Chronic Kidney Disease

Background: Blood urea nitrogen and serum creatinine concentrations only detect a decrease of >75% of renal functional mass. Therefore, there is a need for markers that allow early detection and localization of renal damage.
Hypothesis: Urinary albumin (uALB), C-reactive protein (uCRP), retinol binding protein (uRBP), and N -acetyl-β- d -glucosaminidase (uNAG) concentrations are increased in dogs with chronic kidney disease (CKD) compared with healthy controls and in healthy older dogs compared with young dogs.
Animals: Ten dogs with CKD, 10 healthy young dogs (age 1–3 years), and 10 healthy older dogs (age > 7 years) without clinically relevant abnormalities on physical examination, hematology, biochemistry, and urinalysis.
Methods: Urinary markers were determined using an ELISA (uALB, uCRP, and uRBP) or a colorimetric test (uNAG). Results were related to urinary creatinine (c). The fixed effects model or the Wilcoxon rank sum test were used to compare the different groups of dogs.
Results: uALB/c, uRBP/c, and uNAG/c were significantly higher in CKD dogs than in healthy dogs. No significant difference was found for uCRP, which was not detectable in the healthy dogs and only in 3 of the CKD dogs. Between the healthy young and older dogs, no significant difference was detected for any of the markers.
Conclusion: The urinary markers uALB/c, uRBP/c, and uNAG/c were significantly increased in dogs with CKD compared with healthy controls. Additional studies are needed to evaluate the ability of these markers to detect renal disease before the onset of azotemia.     

浅析大树移植技术

本文从大树的选择、移植时间、移植前的准备工作、栽植后的管理等几个方面阐述了大树移植技术。     

A model of necroptosis in human renal tubular epithelial cells

AIM:To make a model of necroptosis in human renal tubular epithelial HK-2 cells. METHODS:To induce necroptosis, HK-2 cells were treated with tumor necrosis factor α (TNF-α) followed by ATP depletion, and benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (zVAD-fmk) was added to block the activity of caspase-8. The morphological changes of the cells were observed under light microscope and electronic microscope.The cell viability was detected by CCK-8 assay, and the marker of necroptosis was analyzed by Western blotting. RESULTS:In the cells treated with TNF-α followed by zVAD-fmk and antimycin A for 1 h, the morphological changes including the cell and organelle inflation, and membrane fragmentation, with a large amount of autophagysome, were observed.However, these abnormalities were markedly attenuated after treatment with Nec-1. Meanwhile, the cell viability was also significantly improved after using Nec-1. No similar variation was observed in other groups. In addition, the expression of LC3-II was significantly decreased in Nec-1+TNF-α+zVAD-fmk+ antimycin A (1 h) group compared with control group. CONCLUSION: TNF-α stimulation and energy depletion induce necroptosis in renal tubular epithelial cells.Nec-1 inhibits necroptosis in a caspase-independent pathway, and may have therapeutic potential to prevent and treat renal ischemia injury.     

奶牛肾小管上皮细胞的原代培养和鉴定

为建立奶牛肾小管上皮细胞的原代培养、传代以及鉴定方法,本研究将肾脏皮质剪碎,并网筛过滤选取肾小管后分别进行直接培养、胶原酶1消化培养、胰蛋白酶消化培养和胰蛋白酶和0．02％EDTA混合消化培养,分别应用上述四种方法进行原代培养和传代培养,然后采用CK-18、PCK和Vimentin三种蛋白进行免疫组化鉴定奶牛肾小管上皮细胞。结果表明,采用消化培养方法都能够得到奶牛肾小管上皮细胞,但是胶原酶1消化得到的活细胞更多,能更快地成功培养奶牛肾小管上皮细胞。胶原酶1消化法是培养奶牛肾小管上皮细胞的理想培养法。     

猪精原干细胞体外分离培养及移植的研究进展

精原干细胞（spermatogonial stem cells,SSCs）位于睾丸曲细精管基底小室内,因其具有持续的自我更新能力和分化为精子的能力,被认为是雄性哺乳动物体内唯一能将自身遗传物质传递给后代的成体干细胞。对SSCs的研究已成为干细胞学科研究的热点之一,但目前SSCs的研究多集中于啮齿类动物,而猪SSCs（porcine SSCs,pSSCs）的研究进展相对落后,主要是由于存在以下几个问题：睾丸中pSSCs本身数量极少,且缺乏特异的分子标记;pSSCs的分离纯化技术仍不成熟;pSSCs体外培养体系仍不够完善。这些问题的存在导致pSSCs分离纯化效率低,并且难以在体外长期稳定培养及传代,以至于pSSCs的相关机理研究缺乏稳定的材料,为其体外研究及应用带来了不便。基于以上现状,文章总结了pSSCs体外分离培养及移植的研究进展,详细介绍了pSSCs的分子标记、分离纯化和体外培养的相关方法,以及pSSCs移植技术的研究现状,旨在为pSSCs研究提供参考,以期加快pSSCs在猪的遗传育种与繁殖领域以及男性生殖医学领域的研究和应用。     

鱼类细胞核移植研究的进展及意义

细胞核移植技术是目前生物技术领域研究的热点之一,其科学意义和应用价值重大。文章介绍了中国和世界上鱼类细胞核移植的研究成果及其最新进展;综述了鱼类细胞核移植的基本程序、方法和影响因素,包括胞质受体的准备、核供体的制备、核移植和核质杂种的培养。并对鱼类细胞核移植的应用、现存问题及应用前景进行了讨论。