广西巴马地区居民血清中叉头框蛋白O3A基因mRNA及蛋白水平的研究

目的研究广西巴马地区居民血清中叉头框蛋白O3A(forkhead box protein O3A,FOXO3A)基因mRNA及蛋白表达水平的差异,探讨其与家庭聚集性长寿现象的关系。方法于2011年5月,在广西巴马地区抽取具有3~4代直系亲属的家庭,分为长寿家族史组(137例)和无长寿家族史组(91例),测定血清中FOXO3A mRNA及蛋白的表达水平。结果长寿家族史组人群血清FOXO3A mRNA的表达水平是无长寿家族史组的1.21倍,差异有统计学意义(P0.05)。长寿家族史组和无长寿家族史组人群血清FOXO3A蛋白的表达水平间比较,差异无统计学意义(P0.05)。长寿家族史组居民血清中FOXO3A mRNA的表达水平与其蛋白的表达水平之间未见相关关系(P0.05);无长寿家族史组居民血清中FOXO3A mRNA的表达水平与其蛋白表达水平呈负相关(P0.05)。结论广西巴马地区居民血清FOXO3A mRNA的表达水平可能与当地家庭聚集性长寿现象有一定关联,尚未发现FOXO3A蛋白水平与家庭聚集性长寿现象的关联性,需进一步扩大样本量进行验证。     

Segmental innervation of the Göttingen minipig hind body. An electrophysiological study

The Göttingen minipig is being used increasingly in biomedical research. The anatomical structure of the porcine peripheral nervous system has been extensively characterized, but no equivalent to the dermatome map, which is so valuable in human neurophysiological research, has been created. We characterized the medullar segmental skin and muscle innervations of the minipig hind body, using neurophysiological methodology. Six adult minipigs underwent unilateral laminectomy from L2 to S3, exposing the nerve roots. The skin of the hind part of the body was divided into 36 predefined fields, based on anatomical landmarks for consistent reproducibility. We recorded the evoked potential in each exposed nerve root L2‐S3 for cutaneous stimulation of each skin field, mapping the sensory innervation of the entire hind body. We subsequently recorded the motor response in seven predefined muscles during sequential stimulation of the L2‐S3 nerve roots. We obtained a clear sensory evoked potential in the nerve roots during stimulation of the skin fields, allowing us to map the sensory innervation of the minipig hind body. Neurophysiological data from skin stimulation and muscle recordings enabled us to map the sensory innervation of the Göttingen minipig hind body and provide information about muscular innervation. The skin fields were sensory innervated by more than one root. The muscles each had one dominant root with minor contribution from neighboring roots. This is consistent with experimental data from human studies.     

The effect of dietary protein intake on immune status in pigs of different genotypes

Little is known about the immune status of different pig breeds and the role of dietary protein intake. The present study, performed on 96 barrows (48 pure-bred Bama mini-pigs (fatty genotype) and 48 Landrace pigs (lean genotype)) randomly assigned to two dietary treatment groups (low- or adequate protein diet), with twenty-four24 individuals per treatment), evaluated the levels of immunoglobulins and cytokines in serum, and intestinal and splenic innate immunity on the two breeds. Data showed significant interactions between breed and diet for adaptive and innate immunity. Throughout the experiment, Landrace pigs showed significantly higher intestinal immune responses than Bama mini-pigs, and an adequate provision of dietary protein improved the expression of specific antibodies in early growth phases. These finding have important implication for improving the immune status of Landrace pigs, which are a major source of protein for humans.

Abbreviations: BW: Body Weight; NRC diet: National Research Council diet; GB diet: Chinese conventional diet; CSFV: Classical Swine Fever Virus; IgG: Immunoglobulin G; IgM: Immunoglobulin M; SIgA: Secretory Immunoglobulin A.     

Canalostomy is an ideal surgery route for inner ear gene delivery in big animal model

Abstract

Background: Inner gene therapy offers great promises as a potential treatment for hearing loss.

Aims/objectives: One of the critical determinants of the success of inner ear gene therapy is to find a delivery method which results in consistent transduction efficiency of targeted cell types while minimizing hearing loss.

Material and methods: Surgery was performed only in the right ear of each Bama miniature pig, and the left ear served as a control. The gene delivery to inner ear via round window membrane (RWM) and posterior semicircular canal (PSC) approach was performed with the viral vector AAV1-CMV-GFP.

Results: The gene delivery through RWM and the PSC (canalostomy) is able to perfuse the inner ear.

Conclusions and significance: The easy anatomic identification of the PSC, as to RWM, as well as minimal manipulation of the temporal bone required, make this surgical approach an attractive option for inner ear gene delivery in big animal model.     

Comparison of the osteogenic capacity of minipig and human bone marrow‐derived mesenchymal stem cells

Minipigs are a recommended large animal model for preclinical testing of human orthopedic implants. Mesenchymal stem cells (MSCs) are the key repair cells in bone healing and implant osseointegration, but the osteogenic capacity of minipig MSCs is incompletely known. The aim of this study was to isolate and characterize minipig bone marrow (BM) and peripheral blood (PB) MSCs in comparison to human BM‐MSCs. BM sample was aspirated from posterior iliac crest of five male Göttingen minipigs (age 15 ± 1 months). PB sample was drawn for isolation of circulating MSCs. MSCs were selected by plastic‐adherence as originally described by Friedenstein. Cell morphology, colony formation, proliferation, surface marker expression, and differentiation were examined. Human BM‐MSCs were isolated and cultured from adult fracture patients (n = 13, age 19–60 years) using identical techniques. MSCs were found in all minipig BM samples, but no circulating MSCs could be detected. Minipig BM‐MSCs had similar morphology, proliferation, and colony formation capacities as human BM‐MSCs. Unexpectedly, minipig BM‐MSCs had a significantly lower ability than human BM‐MSCs to form differentiated and functional osteoblasts. This observation emphasizes the need for species‐specific optimization of MSC culture protocol before direct systematic comparison of MSCs between human and various preclinical large animal models can be made. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 30:1019–1025, 2012     

长寿地区成年女性骨密度测量结果评估

目的 检测广西巴马地区20岁～60岁成年女性的骨密度,探讨其骨密度变化的规律.方法 随机选取当地268位20岁～60岁成年女性,按不同年龄分组,每组5岁,共8组.应用韩国生产跟骨超声骨密度测定仪,测定跟骨骨密度.采用SPSS16.0对数据进行分析.结果 268例全部进入分析.(1)女性的峰值骨密度处于36岁～40岁年龄组,峰值组的骨密度高于绝经后组(P＜0.05),且随年龄的增加呈现先升高后下降的趋势.(2)女性的骨密度随年龄的增加而减少,导致骨质疏松症的发病率升高.(3)Pearson相关分析显示,女性的SI和年龄呈现出负相关(r=-0.193,P＜0.01),和身高有显著关系(r=0.157,P＜0.05),并未发现与体重和BMI有关系.结论 广西巴马地区20岁～60岁女性SI值与年龄和身高均有显著相关性,本研究获得的SI值为进一步探讨长寿地区成年女性的骨密度状况提供参考依据.     

Determination of critical size defect of minipig mandible

The critical size defect (CSD) of bone can provide a standard for evaluating the usefulness of bone repair materials or methods. The present study aimed to determine the CSDs of the minipig mandible with and without the periosteum. Ten 18 month‐old female minipigs were used. First, the premolars and the first molar in the right mandible of each minipig were extracted. Two months later, the animals were randomly divided into two groups, with six animals in group A and four in group B. In group A, bone segments of 4, 5 and 6 cm were removed from the right mandible and the neighbouring periosteum was preserved. In group B, bone segments of 1 and 2 cm were removed from the right mandible and the periosteum was removed. The defects were retained with two reconstructive titanium plates. The mandibles underwent X‐ray examinations at 4, 8 and 12 weeks after the operation. The animals were sacrificed at 12 weeks after the operation, and specimens were evaluated by gross and histological examinations. For defects retained by two reconstructive plates, the CSDs in the minipig mandible models with and without periosteum were determined as 6 and 2 cm, respectively. Copyright © 2009 John Wiley & Sons, Ltd.     

体外培养版纳微型猪近交系骨髓基质干细胞的生物学特性

目的　分离培养成年版纳微型猪近交系骨髓基质干细胞并探讨其体外生物学特性。方法　成年版纳微型猪骨髓基质干细胞通过密度梯度离心法分离获得 ,用含 15 %小牛血清的 DMEM在 37℃ ,5 % CO2 条件下培养 ;通过克隆生长特性、特异抗原表达和成骨分化能力鉴定其干细胞特性。结果　版纳微型猪近交系骨髓分离获得的基质干细胞具有多种形状 ,但主要为梭形。其具有很强的增殖能力 ,在体外培养条件下可以观察到明显的克隆样生长 ;表达 SH2、SH3、SH4、SB10和 SB2 1等骨髓基质干细胞的特异标记 ;经成骨添加剂诱导培养之后能分化为成骨细胞 ,碱性磷酸酶活性增强并具有细胞外基质矿化能力。结论　成年版纳微型猪近交系骨髓来源的基质干细胞具有干细胞的一般生物学特性     

FOXO3A基因多态性与广西巴马县长寿现象的相关性研究

目的了解FOXO3A基因8个SNPs位点在广西巴马地区不同人群中的频率,探讨FOXO3A基因多态性与巴马地区长寿现象的相关性。方法采用TaqMan探针实时荧光PCR技术对广西巴马长寿地区177例长寿老人(年龄90～110岁,长寿组)以及巴马非长寿地区148例健康成年人(年龄48～89岁,对照组)的FOXO3A基因进行基因分型。结果长寿组中FOXO3A基因rs2764264、rs9400239、rs13217795、rs2802288、rs2802292位点的小等位基因频率高于对照组,差异均具有统计学意义(P0.05)。通过连锁不平衡构建两组人群的FOXO3A基因8个SNPs位点的单体型,发现了GA、AG、TTTGTC、GTCGCT、TCTGTC5个主要的单体型(频率5%)。与对照组比较,长寿组中单体型AG的频率较高,单体型TTTGTC的频率较低,差异均有统计学意义(P0.05)。结论FOXO3A基因5个SNPs及2个基于连锁不平衡构建的单体型可能与巴马长寿地区的长寿现象有一定的联系。