Comparison of the pig and feline models for full thickness corneal transplantation

Purpose The goal of this study was to report on the advantages and limitations of the pig and feline models for experimental in vivo corneal transplantation. Methods Ten healthy domestic pigs and ten healthy cats were used. Full thickness penetrating keratoplasty was performed using autologous (eight cases), allogeneic (seven cases) or human xenogeneic (three cases) tissue. In two other cases, the inflammatory response to partial thickness trephination (without transplantation) was evaluated. Eyes were assessed daily before and after surgery by slit‐lamp, pachymetry, and tonometry. A transparency score ranging from 0 (opaque graft) to 4 (clear graft) was used, based on the slit‐lamp examination. Optical coherence tomography, histology, and electron microscopy were performed postmortem. Results In the pig, the mean (±SD) transparency score for the eight full thickness grafts was 0.88 ± 0.99, ranging from 0 to 3. In the feline model, the mean transparency score for the seven uncomplicated grafts was 3.93 ± 0.19, ranging from 3.5 to 4. Both negative controls without endothelium remained opaque at all time. Intraoperative tendency for iris incarceration into the wound, rapid corneal swelling, suture cheese wiring, and postoperative intraocular inflammation were the main factors jeopardizing the functional success of the corneal transplant in the pig model. Conclusion Suboptimal functional results were obtained after full thickness corneal transplantation in the pig model, while in the feline model, the same protocol yielded uneventful surgeries and clear transplants, with functional results similar to those achieved in human subjects.     

供体年龄对德国肉用美利奴羊超数排卵及胚胎移植效果的影响

提高绵羊体内胚胎生产的效率对促进优良品种扩繁及良种推广具有重要意义。为充分利用不同年龄段良种德国肉用美利奴羊,试验选用青年(18月龄,配种未受胎)、老龄(60~72月龄,经产)以及幼龄母羊(7月龄,性成熟前)做供体,用改进的程序进行超数排卵,研究供体年龄与超数排卵和胚胎移植效果的关系。结果表明:①青年、老龄组母羊超排后只均可用胚数分别为10.9枚、10.3枚,显著高于幼龄母羊的6.0枚。青年、老龄组母羊的胚胎经移植后受胎率分别为71.6%、58.3%,显著高于幼龄组的41.7%。②幼龄、青年组回收的胚胎中囊胚所占比例显著高于老龄组,说明幼龄、青年组供体的胚胎发育速度显著快于老龄供体。③青年、老龄组母羊超排前预注小剂量FSH,卵巢黄体数目明显增多,说明能提高超排效果。本试验优化了德国美利奴羊的超排程序,证明年龄显著影响德国美利奴羊的超排效果及胚胎质量,为进一步提高绵羊胚胎移植效率提供了参考依据。     

鱼类精原干细胞的研究进展

鱼类精原干细胞是鱼类精子发生的起始细胞,是一群具有自我更新和分化潜能的细胞.其独具的生物学特性,使其在干细胞生物学、遗传资源保存和转基因动物等研究领域具有重要价值.就鱼类精原干细胞的生物学特性、培养鉴定、分化潜能、移植和冷冻保存方面做一简要综述,以期为精原干细胞的研究提供借鉴.     

Seasonal influence of climate manipulation on microbial community structure and function in mountain soils

Microbial communities drive soil organic matter (SOM) decomposition through the production of a variety of extracellular enzymes. Climate change impact on soil microbial communities and soil enzymatic activities can therefore strongly affect SOM turnover, and thereby determine the fate of ecosystems and their role as carbon sinks or sources.To simulate projected impacts of climate change on Swiss Jura subalpine grassland soils, an altitudinal soil transplantation experiment was set up in October 2009. On the fourth year of this experiment, we measured microbial biomass (MB), microbial community structure (MCS), and soil extracellular enzymatic activities (EEA) of nine hydrolytic and oxidative extracellular enzymes in the transplanted soils on a seasonal basis.We found a strong sampling date effect and a smaller but significant effect of the climate manipulation (soil transplantation) on EEA. Overall EEA was higher in winter and spring but enzymes linked to N and P cycles showed higher potential activities in autumn, suggesting that other factors than soil microclimate controlled their pool size, such as substrate availability. The climate warming manipulation decreased EEA in most cases, with oxidative enzymes more concerned than hydrolytic enzymes. In contrast to EEA, soil MB was more affected by the climate manipulation than by the seasons. Transplanting soils to lower altitudes caused a significant decrease in soil MB, but did not affect soil MCS. Conversely, a clear shift in soil MCS was observed between winter and summer. Mass-specific soil EEA (EEA normalized by MB) showed a systematic seasonal trend, with a higher ratio in winter than in summer, suggesting that the seasonal shift in MCS is accompanied by a change in their activities. Surprisingly, we observed a significant decrease in soil organic carbon (SOC) concentration after four years of soil transplantation, as compared to the control site, which could not be linked to any microbial data.We conclude that medium term (four years) warming and decreased precipitation strongly affected MB and EEA but not MCS in subalpine grassland soils, and that those shifts cannot be readily linked to the dynamics of soil carbon concentration under climate change.     

小梁切除术联合羊膜植入和干扰素α-2b治疗难治性青光眼

目的 :观察小梁切除术联合同种异体羊膜植入和术眼球结膜下注射干扰素 α- 2 b治疗顽固性青光眼的疗效。方法 :对 1 3例 (1 3只眼 )顽固性青光眼行小梁切除术联合羊膜移植于巩膜床 ,分别于术时、术后 1 d、3 d、7d、1 4 d术眼球结膜下注射干扰素α- 2 b 50万单位。术后随访 6～ 1 5个月 ,平均 1 0 .5个月。观察术后阻止瘢痕性滤过泡形成的效果、降眼压作用、手术成功率及视力变化。结果 :1 3只眼中 ,形成功能性滤过泡 1 1只眼 ,无功能滤过泡 2只眼 ,术后 3个月眼压从术前的 (36.42± 5.64) mm Hg降低到 (1 5.73± 2 .99) mm Hg(t=1 1 .465 ,P<0 .0 1 )。术后1 2个月末 ,手术的成功率为 79.8% (寿命表法 )。结论 :小梁切除术联合羊膜植入与干扰素α- 2 b注射能提高成功率 ,且安全、可靠。     

Canine Haematopoietic Chimerism Analyses by Semiquantitative Fluorescence Detection of Variable Number of Tandem Repeat Polymorphism

Canine models are successfully applied to the study of haematopoietic stem cell transplantation (HSCT). Monitoring of haematopoietic donor/recipient chimerism is of major significance in detecting and quantifying engraftment or graft rejection of the donor-derived haematopoietic cells after transplantation. Radioactive analyses of polymorphic microsatellite markers are commonly used for chimerism analyses. We describe an improved, non-isotopic method that is based on the analysis of microsatellite markers in donor and recipient cells using capillary electrophoresis and fluorescence detection. Artificial mixtures of donor and recipient DNA that were generated from peripheral blood mononuclear cells from dog leukocyte antigen-identical siblings were used to analyse the sensitivity of the assay. DNA from dogs that had received HSCT were also analysed in order to demonstrate the feasibility of the method in vivo. For chimerism analyses, six different microsatellite loci were systematically amplified using fluorescent PCR primer. The fluorescent polymerase chain reaction products were separated by capillary electrophoresis using POP4 on a 310 ABI Prism Genetic Analyzer. After electrophoresis, fluorescence signals were automatically sized and quantified using GeneScan software. The method described provides an accurate assessment of haematopoietic chimerism in the canine model with significantly reduced hands-on time compared to conventional gel electrophoresis.     

Comparison of 3 techniques for ureteroneocystostomy in cats

OBJECTIVE: To compare 3 techniques for ureteroneocystostomy in cats. STUDY DESIGN: Experimental surgical study. ANIMALS: Fifteen adult cats. METHODS: Cats (15) had ureteroneocystostomy with ureteronephrectomy of the contralateral kidney: 5 cats had an intravesical mucosal apposition technique (modified Leadbetter-Politano; intravesical-MA group), 5 cats had extravesical ureteroneocystostomy (modified Lich Gregoir) using a simple continuous suture pattern (extravesical-SC group) and 5 cats had an extravesical technique using a simple interrupted suture pattern (extravesical-SI group). Renal function was evaluated by measuring serum creatinine concentration. Ultrasonographic assessment of the kidney and ureteroneocystostomy site was performed the day after surgery, twice weekly for 3 weeks and once weekly for the remainder of the study. Cats were euthanatized 50 days after surgery. The kidney and ureter removed at surgery, the remaining kidney, ureter, ureteroneocystostomy site, and bladder were examined histologically. RESULTS: Two extravesical-SC cats were euthanatized because of azotemia and uroabdomen, and 1 died acutely at day 4 for unknown reasons. In the intravesical-MA and extravesical-SI cats, the serum creatinine concentration increased after surgery, peaking at a mean (+/-SD) of 9.4+/-2.4 mg/dL and 4.9+/-3.3 mg/dL on day 3, and decreasing to 3.4+/-5.7 mg/dL and 1.5+/-0.4 mg/dL on day 7, respectively. The extravesical-SI technique was associated with consistently lower serum creatinine concentrations for the first week after surgery compared with the other techniques. The mean serum creatinine concentration was within the reference range in cats in the intravesical-MA and extravesical-SI groups by days 10 and 5, respectively. Renal pelvic dilatation occurred in all cats but resolved more rapidly in cats after extravesical techniques. There was no significant difference in serum creatinine concentrations or renal pelvic dilation between the intravesical-MA and extravesical-SI techniques. Bladder mass height at the anastomosis site was significantly larger and persisted for longer with intravesical-MA technique. CONCLUSION: An extravesical-SI technique is seemingly the choice for ureteroneocystostomy in cats with undilated ureters. Renal pelvic dilation on ultrasound examination should be expected after ureteroneocystostomy in cats. CLINICAL RELEVANCE: An extravesical ureteroneocystostomy technique using a simple interrupted pattern for anastomosis should be considered in cats undergoing renal transplantation.     

鳜→鲤的细胞核移植

本文报道用细胞核移植的方法,获得了亲缘关系较远的不同目鱼类之间的核一质杂种鱼胚胎.即以鳜鱼的囊胚细胞核作供体,以鲤鱼的成熟未受精去核卵细胞质作为受体进行核移植试验,使囊胚细胞核发生分裂,从而启动卵细胞质也发生分裂,导致重新进行个体发育.在获得的112个正常发育的核移植卵中,发生卵裂的107个,占95.5%发育至囊胚期的61个,占54.5%,发育至原肠期的6个,占5.4%,发育至神经胚期和胚孔封闭期的各1个,分别占0.9%.鳜→鲤核质杂种胚胎类似鲤鱼同期胚胎,其早期发育速度与鲤鱼相似而比鳜鱼快.各杂种胚胎发育期的胚盘形状及大小均类似鲤鱼,大于鳜鱼同期胚盘,且胚盘细胞较多,但比鲤鱼胚盘细胞小,这是鳜→鲤核质杂种胚胎发育的一个显著特点.作者认为核-质杂种胚胎的性状发育,受核受体物种的母性影响控制.     

桂花大树移植技术研究

在城市追求快速绿化的形势下,大树移植已成为一种常态,而集"绿化、美化、香化"于一身的桂花,更是成为了市场宠儿,为更好地利用好这些日益稀缺的桂花大树资源,掌握桂花大树移栽技术,对桂花大树移植的树源选择、起苗、运输、栽植与养护等多个环节进行了详细、系统的研究,以对市场的桂花大树移植提供技术参考。     

以鲁西黄牛为受体的高产奶牛胚胎移植研究

以自然发情的鲁西黄牛为受体,移植经性控技术生产的高产奶牛7日龄雌性冷冻胚胎,共移植合格受体牛5 516头,移植后60～80d进行直肠孕检,怀孕2 228头,移植准胎率为40.39％;共繁殖奶牛犊牛1346头,其中母牛1 279头,公牛67头,胚胎性别控制率为95.02％.左侧与右侧卵巢排卵移植准胎率分别为35.14％和43.15％,差异极显著(P＜0.01);春季与秋季的移植准胎率分别为36.93％和43.53％,秋季明显高于春季,差异极显著(P＜0.01);黄牛不同黄体级别A级、B级、C级的准胎率分别为39.16％、41.70％和33.42％,A级与B级移植准胎率差异不显著(P＞0.05),B级与C级移植准胎率差异极显著(P＜0.01);移植单胚与双胚的受体牛准胎率分别为40.73％和38.24％,差异不显著(P＞0.05).