牛胚胎移植试验报告

2000年9月至2003年9月,在大通县青林、多林、逊让、向化等9个乡镇开展了安格斯牛、荷斯坦、皮埃蒙特牛冷冻胚胎移植工作。经过对298头当地黄牛母牛及杂交母牛进行了诱导同期发情,结果获得93.29%的同期发情率和38.13%的黄体合格率。对于黄体合格的106头母牛进行了安格斯、荷斯坦、皮埃蒙特牛冷冻胚胎移植,获得移植妊娠率42.5%。     

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.     

不同粪菌悬液处理方式对猪粪菌移植过程中细菌活性及活菌组成的影响

旨在探究不同粪菌液处理方法对其菌群活性及存活菌群物种组成的影响,为猪粪菌移植（FMT）的优化应用提供参考。本研究共设置7个试验组制备粪菌液：静置制备新鲜粪菌液组（FS）、静置制备粪菌液于-80℃冷冻1周组（FFS）、离心制备新鲜粪菌液组（FC）、离心制备粪菌液于-80℃冷冻1周组（FFC）、液氮冷冻粪便24 h静置制备粪菌液组（NS）、甘油+液氮冷冻粪便24 h静置制备粪菌液组（GNS）和液氮冷冻粪便24 h静置制备粪菌液于-80℃冷冻1周组（NFS）,在37℃厌氧培养箱中平板培养48 h后,进行菌落计数。从FS、FFS、FC和FFC 4组中根据菌落形态随机挑取87个单菌落纯化后进行16S rRNA基因全长测序。结果表明,静置和离心两种方法制备新鲜粪菌液时,粪菌活性存在显著差异,离心组显著低于静置组（P<0.05）。但冷冻粪便使用静置和离心两种方法制备的菌悬液,其粪菌活性无显著差异（P>0.05）。无论是静置还是离心方法制备粪菌液,新鲜和冷冻两种不同处理方式下的粪菌活性均无显著差异（P>0.05）,且对活菌的组成也无显著影响。使用静置法制备粪菌液时,FS、NS、GNS和NFS组间粪菌活性无显著差异（P>0.05）。综上表明,静置方法制备粪菌液时,超低温冷冻保存对粪菌活性及其活菌组成均无显著影响,这为大样本、长距离猪粪菌移植技术的应用提供了重要试验参考。     

犬造血干细胞移植的作用机制及应用研究进展

造血干细胞(hematopoietic stem cell, HSC)位于造血系统的顶端, 是造血系统中唯一具有多能性和自我更新能力的细胞, 可以分化为各种功能的血细胞, 维持血液系统的建立和动态平衡, 是目前研究最为透彻、临床应用最为成熟的成体干细胞。造血干细胞的这些重要特性, 使得基于造血干细胞在治病机制研究和临床应用中的研究取得了很大进展。以造血干细胞为基础的再生医学治疗是目前治疗恶性血液病和遗传病的首选方法, 如造血干细胞治疗犬白细胞黏附缺陷综合征、犬遗传性贫血、犬淋巴瘤、犬白细胞减少症、犬X连锁严重联合免疫缺陷病等, 但由于白细胞抗原匹配的供体稀缺、可获得的造血干细胞数量有限等原因, 无法满足临床需求, 因此, 如何通过体外培养获得满足临床需要的造血干细胞成为近年来学者们研究的热点问题。作者参照现有研究成果对造血干细胞的来源和体外分离培养、治病机制进行系统的描述, 并对造血干细胞移植治疗遗传性溶血性贫血、白细胞黏附缺陷综合征、淋巴瘤、白细胞减少症、X连锁严重联合免疫缺陷病的临床研究进行综述, 以期为今后将造血干细胞广泛应用于临床治疗提供参考依据。     

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.     

软枣猕猴桃试管苗驯化移栽技术的研究

[目的]探索软枣猕猴桃试管苗移栽后的最宜生长条件,提高成活率,缩短成活时间。[方法]通过测试不同炼苗季节、预处理方法、栽培基质、温度、湿度、光照对试管苗成活、移栽效果影响,得出最佳移栽炼苗程序。[结果]将生长健壮,苗高5cm以上的组培苗移至室温光照2000-3000lx,10h/d的栽培条件下经4d炼苗后,清水洗苗,蘸生根粉移栽至沙土∶黄棕壤∶腐叶土∶园土(1∶1∶1∶1)的基质中盆栽、套袋后置于棚高50cm的小拱棚内,拉2层遮阴网（透光率40-50%,温度20-24℃,土壤含水量80-90%）培养出的幼苗,成活率高达92%,叶数7,整体生长整齐,是软枣猕猴桃最佳驯化移栽方式。[结论]实验所得方法可为大田生产提供基础,极大程度上提高移栽成活率,缩短成活时间。     

MicroRNA-145 inhibits epithelial-mesenchymal transition in human renal cancer A-498 cells by ADAM28

AIM: To investigate the inhibitory effect of microRNA-145 (miR-145) on epithelial-mesenchymal transition (EMT) in renal cancer A-498 cells. METHODS: The A-498 cells were transfected with miR-145 mimics (M145) and mimic negative control(MNC), which served as M145 group and MNC group, respectively. Mock control (MC) group was set up using untreated A-498 cells. The expression level of miR-145 in each group was detected by RT-qPCR. Transwell assay was used to detect the invasion ability of the cells. The protein expression of vimentin, E-cadherin and ADAM28 was determined by Western blot. Bioinformatic method was used to predict the target genes of miR-145. Antagonistic effect of ADAM28 over-expression on the inhibition of EMT by miR-145 was detected by Western blot. The relationship between miR-145 and ADAM28 was analyzed by dual-luciferase reporter assay. RESULTS: The expression level of miR-145 in M145 group was significantly up-regulated than that in MC group (P<0.05). The number of invasive cells in M145 group was 12.78±3.37, which was significantly lower than that in MC group (P<0.05). ADAM28 may be the target gene of miR-145. Compared with MC group, the protein expression of vimentin and ADAM28 in M145 group was significantly decreased (P<0.05), while the protein expression of E-cadherin was significantly increased (P<0.05).After ADAM28 over-expression, the protein expression of vimentin in the A-498 cells of M145 group was significantly increased (P<0.05), and the protein expression of E-cadherin was significantly decreased (P<0.05). The results of dual-lucife-irasei reporter assay showed that ADAM28 was a downstream target gene of miR-145. CONCLUSION: miR-145 may inhibit the expression of EMT-related proteins through the downstream target gene ADAM28 and inhibit the EMT process of renal cancer A-498 cells.     

水牛卵母细胞去核方法的比较

水牛卵母细胞体外成熟22h后，分别用盲吸法、点击法和纺锤体成像系统（Spindle—View System）去核。结果，三者的去核率分别为65．1％、81．8％和95．0％，差异极显著（P〈0．01）。以水牛胎儿成纤维细胞为供体，通过核移植构建的重构胚的融合率、分裂率和囊胚发育率，在上述3种方法之间均没有显著差异（P〉0．05）。认为点击法是一种较为简单、实用和有效的去核方法。     

香樟大树移植及养护技术措施

根据香樟大树移植程序论述了其移植技术,并从多个角度描述了香樟大树移植后的养护技术措施。