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。     

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.     

Cellular proliferation in the nasal septal cartilage of juvenile minipigs

The growth of the nasal septal cartilage is believed to be a driving force of midfacial growth. Cellular proliferation is an important contributor to growth of the cartilage, but this factor has been rarely investigated. The current study was undertaken to assess the proliferation and cellular density in the septal cartilage of fast-growing juvenile minipigs. Six minipigs averaging 4.4 ± 1 months old were injected with 5′-bromo-2′-deoxyuridine (BrdU), a thymidine analog, 24 h before death. The septal cartilage was sectioned in the coronal plane and reacted for BrdU. The proliferative index (number of BrdU-positive chondrocytes/total number of chondrocytes) and cellular density (number of cells mm⁻²) of various locations of the septum were measured and compared in order to determine overall proliferation rate and whether regional variations in proliferative activity and cellular density are present. To provide a time perspective to the problem of midfacial growth, the lengths of the nasal bone and the palate were measured in a collection of 61 dry skulls of minipigs aged 1–8 months. Results showed that the septal chondrocytes were proliferating at a surprisingly high rate (∼21%). The proliferative index was higher in the ventral and middle compared with the dorsal locations, and in the central cartilage compared with the perichondrium. No difference in proliferative index was found between the anterior and posterior parts of the septum. Cellular density was higher in the perichondrium than in the central cartilage. Within the central cartilage there was a trend for higher cellular density anteriorly. In conclusion, the rapidly growing midface of juvenile minipigs is associated with a high rate of septal proliferation, especially in the ventral half of the cartilage.     

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.     

西藏小型猪IGF-1基因的克隆测序分析

目的 克隆西藏小型猪的肝脏组织中的IGF-1基因的cDNA序列,并与pubmed中查询到的猪cDNA进行比对分析。方法 本研究提取了西藏小型猪肝脏组织的总RNA,应用RT-PCR技术扩增了IGF-1基因的cDNA序列,将扩增出的片段克隆到pMD18-T载体上,构建重组质粒pMD18-T-IGF-1,进行测序分析。结果 首次克隆出了西藏小型猪肝脏组织中的IGF-1的cDNA序列,获得了大小为567bp长的片段,编码了186个氨基酸,与pubmed中查询到的猪（NM_214256.1）的IGF-1基因高度同源,比对序列发现,在440bp、455bp处发生了G→A、C→T的突变,该位点的突变引起相应编码氨基酸的变化,分别由组氨酸变成了精氨酸、亮氨酸转变成了丝氨酸。结论 本研究为西藏小型猪的生长发育机制研究提供了分子学依据,两个位点的突变引起的氨基酸的改变是否是导致西藏小型猪矮小的原因,需要进一步论证。     

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.     

高温高湿环境下战创伤创面模型的建立

目的建立高温高湿环境下猪战伤后创面的动物模型,以供对爆炸伤引起的战伤慢性创面病理生理以及修复治疗进行研究。方法选用藏香猪8头,采用KTY-08型瞬发纸质雷管的爆炸源模拟爆炸冲击波,制作猪的两侧臀部战创伤创面,共计16个创面。致伤后随机分为两组,即常温常湿组（对照组）：气温（Ta）（22．0±0．5）℃,相对湿度（RH）50％;高温高湿组：Ta（39．0±0．5）℃,RH8I）％～85％。每组4只,观察伤后不同时间段创面的大体、病理切片变化及创面愈合时间。结果创面大小平均为（6．0±0．9）cmX（6．5±1．2）cm×（1．5±0．3）cm,12h后高温高湿组较常温常湿组创面明显扩大,有感染征象,第21天时创面愈合率为0,第28天时愈合率为12．5％（1／8）。常温常湿组24h后出现明显感染,第21天后创面完全愈合,愈合率100％。结论藏香猪臀部战创伤创面模型创面面积大,未伤及骨组织,动物可长期存活,且皮肤结构及创面愈合与人类相似,是高温高湿环境下良好的战创伤创面动物模型。     

巴马小型猪在医学研究中的应用进展

广西巴马小型猪作为国内小型猪主要品种之一,具有遗传性稳定、多产、体重较小、体表多覆白毛等特征,组织器官及生化指标与人类相似,已越来越广泛地应用于医学研究领域：猪的心脏解剖与生理特点与人类高度相似,已被广泛应用于心血管系统研究中,在我国,巴马小型猪被用来构建心肌缺血、卵圆孔未闭等心血管疾病模型;猪具有杂食性及与人相似的脂质代谢,可用于研究内分泌疾病,巴马小型猪已用于糖尿病动物模型建立及其遗传易感性、并发症的防治研究等;巴马小型猪的消化系统与人类相似,利用此特点已建立阻塞性慢性胰腺炎、结肠穿孔、胆肠吻合等消化系统疾病模型;巴马小型猪除头、尾外,体表覆以白毛,这一特点使其成为研究皮肤创伤、烧伤修复等的理想动物;小型猪的牙齿解剖结构与人类相似,口裂大,可作为口腔医学研究中的理想动物,巴马小型猪已用来建立牙髓坏死模型及对上颌扩弓方式的研究;类似人的解剖、生理、病理使其成为较为适合的异种移植供体。在中医药研究方面,已用巴马小型猪分别建立了肝脏、脾脏、股动、静脉出血及头颈恶性肿瘤放疗后的腮腺损伤动物模型以研究中药制剂的疗效及机制。     

两种麻醉方法用于小型猪肺动脉高压模型建立的对比研究

目的对比两种麻醉方法用于小型猪肺动脉高压模型建立的效果。方法选择24头广西巴马香猪,随机分为A组(静脉复合腹腔注射组)(n=12)与B组(静吸复合麻醉组)(n=12),比较两种麻醉方法的麻醉效果及死亡率。结果 A组优9例、良3例、差0例、麻醉死亡0例,死亡率为0;B组优8例、良2例、差0例、麻醉死亡2例,死亡率为16.7%。两组组间麻醉效果比较差异无统计学意义(Z=-0.652,P=0.515),死亡率B组高于A组。结论在一些短小的小型猪外科手术中,静脉复合腹腔注射可使麻醉过程更平稳,是一种安全、有效、可行的麻醉方法。