Pathological Changes of Acute Immune Hepatitis in Mini Pigs

ＰａｔｈｏｌｏｇｉｃａｌＣｈａｎｇｅｓｏｆＡｃｕｔｅＩｍｍｕｎｅＨｅｐａｔｉｔｉｓｉｎＭｉｎｉＰｉｇｓＹｉｎｇＧｕｏｑｉｎｇ（殷国庆）ＬｉｎＨｏｕｊｉ（林厚基）ＬｉＪｉｅｑｕａｎ（李解权）ＮａｎｊｉｎｇＨｏｓｐｉｔａｌｆｏｒＩｎｆｅｃｔｉｏｕｓＤｉｓ...     

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.     

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。     

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。     

3种腺相关病毒介导的绿色荧光蛋白对西藏小型猪胚胎成纤维细胞的转染效率

目的比较3种不同血清型腺相关病毒(AAV)介导的绿色荧光蛋白对西藏小型猪胚胎成纤维细胞的转染效率。方法原代分离培养西藏小型猪胚胎成纤维细胞并进行形态学鉴定,按照感染复数(MOI)为103、104、105转染传代两次的猪胚胎成纤维细胞,流式细胞学检测不同血清型AAV对西藏小型猪胚胎成纤维细胞的转染效率,并在倒置显微镜下观察转染后绿色荧光的表达强度;同时,应用MTT方法检测不同血清型AAV对西藏小型猪胚胎成纤维细胞的毒性。结果腺相关病毒AAV2-EGFP对PFF的转染效率随MOI值的增加而增高,MOI为103、104、105时,转染效率分别为(33.68±1.18)%、(50.80±2.59)%和(60.08±1.08)%,而其他两种血清型病毒感染效率均低于AAV2-EGFP。3种病毒转染PFF过程中,细胞生长正常,MTT显示各个转染组与对照组(未转染组)在D570处吸光度无显著差异。结论AAV2血清型载体对体外培养的猪胚胎成纤维细胞的感染效率显著高于其他几种血清型,AAV8和AAV9对PFF感染能力极差,筛选AAV2作为西藏小型猪基因打靶的最适病毒载体;同时,3类血清型病毒载体对猪胚胎成纤维细胞均无明显细胞毒性。     

五指山小型猪繁殖性能测定

目的测定封闭群五指山小型猪繁殖性能指标。方法依据相关标准,测定普通级封闭群五指山小型猪种猪繁殖性能,并通过SAS软件进行相关统计分析。结果封闭群五指山小型猪繁殖行为学参数稳定,母猪性成熟早,胎次对窝产仔率、仔猪生存率以及个体重量有一定的影响,但多数统计学差异不显著。相关性分析显示,窝产活仔数与出生个体重间存在负相关,并且随着胎次的增加负相关性增强。结论封闭群五指山小型猪生产繁育性能较高,具有较好的实验动物化价值。仔猪个体发育过程中个体间差异仍有增大的趋势,提示种群仍需进一步选育和纯化。     

五指山小型猪糖耐量试验及血脂分析

目的 确定五指山小型猪的正常糖耐量水平,并检测血脂,筛查糖、脂代谢异常小型猪.方法 选择未去势的五指山小型猪,禁食不禁水12h后按1.2 ml/kg体重的剂量静脉注射50％葡萄糖注射液,并分别在注糖前、注糖后10、30、60、90和120 min采血测定血糖值.同时采取空腹血清送实验室检测血糖（Glu）、甘油三酯（TG）、总胆固醇（TC）、高密度脂蛋白胆固醇（HDL-C）、低密度脂蛋白胆固醇（LDL-C）等生化指标.结果 静脉注糖后血糖水平出现显著性波动,注糖后10 min内血糖水平达到峰值,随后逐渐下降,直至注糖后90 min恢复到给糖前水平.有7头小型猪的糖耐量部分血糖值超过95％置信度上线值,有12头小型猪部分相关的血清生化指标值超过95％置信度上线值.相关性分析发现Glu与TG、TC、HDL-C、LDL-C这4个指标都存在显著的相关性,多因素分析表明TG、HDL-C与Glu显著相关.结论 筛选出糖代谢或脂代谢异常,以及糖耐量减低的小型猪.     

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。     

Early sacral neuromodulation ameliorates urinary bladder function and structure in complete spinal cord injury minipigs

Aims

To determine the effects of early sacral neuromodulation (SNM) and pudendal neuromodulation (PNM) on lower urinary tract (LUT) function, minipigs with complete spinal cord injury (cSCI) were analyzed. SNM and PNM have been proposed as therapeutic approaches to improve bladder function, for example after cSCI. However, further evidence on efficacy is required before these methods can become clinical practice.

Methods

Eleven adults, female Göttingen minipigs with cSCI at vertebral level T11-T12 were included: SNM (n = 4), PNM (n = 4), and SCI control (SCIC: n = 3). Tissue from six healthy minipigs was used for structural comparisons. Stimulation was started 1 week after cSCI. Awake urodynamics was performed on a weekly basis. After 16 weeks follow-up, samples from the urinary bladder were taken for analyses.

Results

SNM improved bladder function with better capacities and lower detrusor pressures at voiding and avoided the emergence of detrusor sphincter dyssynergia (DSD). PNM and untreated SCI minipigs had less favorable outcomes with either DSD or constant urinary retention. Structural results revealed SCI-typical fibrotic alterations in all cSCI minipigs. However, SNM showed a better-balanced distribution of smooth muscle to connective tissue with a trend towards the reduced progression of bladder wall scarring.

Conclusion

Early SNM led to an avoidance of the emergence of DSD showing a more physiological bladder function during a 4 month follow-up period after cSCI. This study might pave the way for the clinical continuation of early SNM for the treatment of neurogenic LUT dysfunction after SCI.