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.     

Effect of "Shen Yan Yi Qi Solution" on renal function and histopathology of rats suffering from an experimental nephrotoxic serum nephritis

AIM: To investigate the therapeutic effects of "Shen Yan Yi Qi Solution", a chinese Medicine, on nephrotoxic serum nephritis in the rat.METHODS: Nephrotoxic serum nephritis model of the rat was set up by the Clarke’s method. RESULTS: Proteinuria and renal insufficiency occurred in all rats after nephrotoxic serum (NTS) injection, but symptoms in treatment group were reduced. At 2nd week, the typical linear deposition of IgG along the glomerular basement membrane (GBM) was observed by fluorescence microscopy. Mesangial hyperplasia and crescent formation in glomeruli were also observed by light microscopy. In treatment group, all the above lesions were alleviated. CONCLUSION: It demonstrated that "Shen Yan Yi Qi Solution" might alleviate the pathological change of glomeruli and improve renal function.     

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).     

常用分离培养肾细胞方法的比较

[目的]采用常用的分离培养肾细胞方法比较分离培养肾细胞,并提出优化方案。[方法]A法:组织块培养法;B法:1%胶原酶消化分离法;C法:5%胰蛋白酶消化法;D法:0.5%胰蛋白酶-0.02%EDTA连续分离法。[结果]A法初期细胞生长不好,1周后生长良好;B法、C法细胞贴壁状况较好,但细胞生长不均一,呈丘陵状生长,细胞产量较少;采用D法消化肾组织块,能够快速获得大量细胞,并且细胞生长整齐、均一,能够满足后续实验的需要。[结论]使用D法分离培养肾细胞优于其他3种方法,试验效果最佳。     

圣诞树组培苗移植试验

运用正交试验的方法研究不同炼苗时间、遮阳网遮光率、基质以及不同移植月份对圣诞树组培苗移植效果的影响.结果表明,炼苗时间为10 d、遮阳网遮光率为70%、基质为泥炭土:黄心土(2∶1)、移植月份为4月的处理圣诞树移植效果最好,成活率可达97.2%,苗高达11.1 cm.     

华北驼绒藜育苗移栽技术研究

干旱地区华北驼绒藜育苗移栽是提高建植成活率的有效途径。利用生根粉、保水剂及清水浸泡及剪根等处理,对不同株龄和移栽不同时期的华北驼绒藜成活率进行了对比试验研究。结果表明,春季移栽成活率为72．6％。秋季为90．3％,药剂处理能够有效提高成活率。