Effects of Donor Cells Treated with Egg Extracts on the Development Competence of Cloned Embryos     

GUO Qin-qin  FAN Zong-xing  Hao Hai-sheng  LIU Yan  ZHAO Xue-ming  ZHU Hua-bin  DU Wei-hua 《中国畜牧兽医》2016,43(2):477-486

Japanese Balck cattle fetal fibroblasts (JBCFF) were induced with Xenopus leavis egg extracts and somatic cell nuclear transfer (SCNT) was carried out with the reprogrammed JBCFF as donor cells in order to investigate their effects on SCNT efficiency.Three samples of egg extracts were acquired from different Xenopus laevis.The protein contents and kinds in extracts were evaluated with BCA Protein Quantification Kit and SDS-PAGE.Concentration of Digitonin to permeabilize JBCFF was optimized and assessed with PI staining.Reprogrammed cells treated with egg extract were used as donor in SCNT.Additionally the reconstructed embryos were activated with ionomycin+6-DMAP and A23187+6-DMAP to compare their effects on the development competence.The protein contents of extracts samples were 56.2255,64.6570 and 71.2158 μg/mL,respectively,the each extract had the same composition about 40-55 and 70-100 ku.The optimal concentration of Digitonin was 7 μg/mL and the permeabilization rate was 55.44%.After extracts treatment and continuous culture for 6-7 d,JBCFF formed well-defined colony structures.No significant composition difference was found in rates of fusion (92.83% vs 96.04%),cleavage (89.64% vs 89.78%) and blastocyst formation (24.06% vs 23.12%) of cloned embryos when the colony cells and JBCFF without extracts treatment were used as donor cells (P>0.05).Similarly,the two activation methods had no significant effect on the developmental competence of cloned embryos (cleavage rate 92.16% vs 92.28%,blastocyst rate 23.21% vs 24.18%).Conclusively,Xenopus leavis,egg extracts could induce JBCFF reprogramming to a low differentiated state.However donor cells with reprogramming partially could not improve the development of cloned embryos and its mechanism requires further research.  相似文献   

Analysis on Differential Expression Genes in Porcine Aortic Vascular Endothelial Cells Induced by Apolipoprotein C Ⅲ     

YUE Yuan  CHEN Hong-yan  WANG Jia-wei  XU Ming-qiang  DING Yu  JIANG Hao  GAO Yan  ZHANG Jia-bao  YAN Shou-qing 《中国畜牧兽医》2016,43(3):585-591

The aim of this study was to investigate the differential expression genes induced by ApoCⅢ,and study the function of ApoCⅢ.Porcine aortic vascular endothelial cells were successfully isolated using enzyme digestion,and then screened the differential expression genes induced by ApoCⅢ using the Solexa high-throughput sequencing technology.The results showed 647 differential expression genes,including 390 up-regulated genes and 257 down-regulated genes.The qRT-PCR results verified that the gene expression results from Solexa sequencing data were reliable.GO and Pathway analysis showed that the function of differential expression genes were related to immune response,cell apoptosis and death.These findings suggested that ApoCⅢ affected the physiological function of porcine aortic endothelial cells by the molecular pathways of inflammation,cell adhesion and apoptosis,which provided a theoretical basis for further understanding the molecular mechanisms of atherosclerosis caused by ApoCⅢ.  相似文献   

Effect of Vitamin E on Bovine Granulosa Cells Apoptosis and Proliferation through Cx43     

REN Shu-qiang  WANG Jia-wei  CHEN Hong-yan  XU Ming-qiang  JIANG Hao  GAO Yan  ZHOU Jian-zhong  ZENG Fan-ming  LI Cheng-sheng  ZHANG Jia-bao  CHEN Cheng-zhen 《中国畜牧兽医》2016,43(3):615-621

The aim of this study was to determine the effect of vitamin E on Cx43,the mechanism and function of vitamin E on bovine granulosa cells apoptosis and proliferation.In this study,granulosa cells were isolated from bovine ovary and cultivated in vitro by adding different concentration of vitamin E (0,25,50,100,200 and 500 μmol/L) for 24 h.After cultured,apoptotic cells were detected by FCM,mRNA expression levels of BCL2/BAX、P53 and Cx43 genes were determined by Real-time PCR and cell proliferation was detected by CCK8.The results showed that compared to control group,100 μmol/L vitamin E could significantly inhibit the apoptosis of granulosa cells (P<0.05).Real-time PCR detection results showed that vitamin E significantly changed the mRNA expression levels of BCL2/BAX,P53,Cx43 genes (P<0.05).Vitamin E could significantly improve granulosa cells proliferation when granulosa cells were treated for 24 and 36 h (P<0.05).The results provided a theoretical basis on further analysis for studing the influence mechanism of vitamin E on oocytes development and maturity,and improvement of female animal reproduction by influencing granulosa cells proliferation and apoptosis.  相似文献   

Screening of Probiotics Inhibiting E. coli K88 in vitro     

WANG Lei  LV Yang  ZHANG Yue  XIE Jia-qian  ZHAO Di  YAN Li-qiong  MEI Hui-min  WANG Lei  YI Dan  CHEN Hong-bo  DING Bin-ying  HOU Yong-qing  WU Tao 《中国畜牧兽医》2016,43(5):1355-1360

The probiotics tested in the experiment were isolated from the intestinal of weaning piglets.The isolated probiotics and E.coli K88 were inoculated into the culture of intestinal porcine epithelial cell-1 (IPEC-1).The activity of lactate dehydrogenase (LDH) in the supernatant was measured after incubating for 2.5 h.At the same time, the probiotics and E.coli K88 were co-cultured in vitro, the number of E.coli K88 was counted and the probiotics which could be resistant to the E.coli K88 were selected 2.5 h later.The results showed that the Lactobacillus casei and Bacillus coagulans could significantly reduce LDH activity (P<0.05), decrease the damage of E.coli K88; Bacillus coagulans could inhibit the growth of E.coli K88.At the same time, Bacillus coagulans could resist high temperature, acid and bile salt.The results showed that Bacillus coagulans strains had great potential as the application of probiotics strains.The methods could be used as a model of screen probiotics which could inhibit the growth of E.coli K88 in vitro.  相似文献   

应用分子靶点药物治疗犬肥大细胞瘤的研究进展     

李格宾  杨紫嫣  林珈好  夏兆飞 《动物医学进展》2016,(2):80-84

犬肥大细胞瘤因其恶性率和转移率都较高,并且常规化疗药物副作用较强,患犬生存期和生存质量显著下降,因此急需新型的药物进行治疗。近几年,治疗肿瘤的分子靶点药物凭借其疗效好、副作用低的特点,受到越来越广泛的关注,特别是在治疗犬肥大细胞瘤的应用中效果显著。论文重点介绍了欧美小动物临床最常用的伊马替尼、托赛拉尼、马赛替尼这3种以异常KIT为靶点的药物及其药物作用机理及药效,最后介绍了由c-kit基因二次突变所导致的耐药问题及其预防措施,期望能对我国小动物临床医师和科研工作者有所启发。  相似文献   

Improved development of somatic cell cloned bovine embryos by a mammary gland epithelia cells in vitro model     

Xiao-ying He  Li-bing Ma  Xiao-ning He  Wan-tong Si  Yue-Mao Zheng 《Journal of veterinary science (Suw?n-si, Korea)》2016,17(2):145-152

Previous studies have established a bovine mammary gland epithelia cells in vitro model by the adenovirus-mediated telomerase (hTERT-bMGEs). The present study was conducted to confirm whether hTERT-bMGEs were effective target cells to improve the efficiency of transgenic expression and somatic cell nuclear transfer (SCNT). To accomplish this, a mammary-specific vector encoding human lysozyme and green fluorescent protein was used to verify the transgenic efficiency of hTERT-bMGEs, and untreated bovine mammary gland epithelial cells (bMGEs) were used as a control group. The results showed that the hTERT-bMGEs group had much higher transgenic efficiency and protein expression than the bMGEs group. Furthermore, the nontransgenic and transgenic hTERT-bMGEs were used as donor cells to evaluate the efficiency of SCNT. There were no significant differences in rates of cleavage or blastocysts or hatched blastocysts of cloned embryos from nontransgenic hTERT-bMGEs at passage 18 and 28 groups (82.8% vs. 81.9%, 28.6% vs. 24.8%, 58.6% vs. 55.3%, respectively) and the transgenic group (80.8%, 26.5% and 53.4%); however, they were significantly higher than the bMGEs group (71.2%, 12.8% and 14.8%), (p < 0.05). We confirmed that hTERT-bMGEs could serve as effective target cells for improving development of somatic cell cloned cattle embryos.  相似文献   

The 5‐lipoxygenase inhibitor tepoxalin induces oxidative damage and altered PTEN status prior to apoptosis in canine osteosarcoma cell lines          下载免费PDF全文

J. P. Loftus  D. Cavatorta  J. J. Bushey  C. B. Levine  C. S. Sevier  J. J. Wakshlag 《Veterinary and comparative oncology》2016,14(2):e17-e30

The 5‐lipoxygenase (5‐LOX) inhibitor tepoxalin has been shown to slow canine osteosarcoma (OSA) tumour xenografts growth, yet the mechanisms are poorly elucidated. Further examination of tepoxalin in canine OSA cell lines shows that tepoxalin treated cells undergo apoptosis through caspase‐3 activation and annexin staining. Interestingly, apoptosis is superseded by an increase in reactive oxygen species (ROS), as measured by activation of dihydrorhodamine 123 and mitosox. This increase in ROS appears to be related to the 5‐LOX inhibitor regardless of cellular 5‐LOX status, and was not observed after treatment with the tepoxalin metabolite RWJ20142. Additionally, 5‐LOX inhibition by tepoxalin appears to increase phosphatase and tensin (PTEN) homolog activity by preventing its alkylation or oxidation. PTEN modification or inhibition allows phosphoinositide‐3 (PI3) kinase activity thereby heightening activation of protein kinase B (AKT) phosphorylation. Our data suggest that off target oxidation and LOX inhibition play roles in the apoptotic response.  相似文献   

Lymphangiosarcoma in 12 dogs: a case series (1998–2013)          下载免费PDF全文

K. M. Curran  C. H. C. Halsey  D. R. Worley 《Veterinary and comparative oncology》2016,14(2):181-190

Lymphangiosarcoma (LAS) is an uncommon malignant neoplasia arising from lymphatic endothelium; little information exists regarding therapy. Single institutional retrospective review for canine LAS histopathology diagnoses over a 15‐year period yielded 12 dogs. Ten dogs were presented for a mass and/or swelling at cervical, trunk or limb regions. Prior to diagnosis, 10 dogs received empiric wound therapy. Cytology performed in 10 consisted of mild inflammation. Survival ranged from 60, 168 and 876 days for three dogs with palliation; 90 days with prednisone in one; 182 days with chemotherapy in one; 240, 267, 487, 630 and 941 days for five receiving surgery; and 574 days for one receiving surgery, radiation and chemotherapy. One dog is alive with recurrence at 243 days following surgery and carboplatin chemotherapy. Clinical improvement existed in LAS dogs receiving multimodal therapies. Early tissue biopsies are recommended for progressive oedematous lesions of unknown origin.  相似文献   

Cytological grading of canine cutaneous mast cell tumours          下载免费PDF全文

Filippo Scarpa  Silvia Sabattini  Giuliano Bettini 《Veterinary and comparative oncology》2016,14(3):245-251

A cytological grading for mast cell tumours (MCTs) would be highly desirable, allowing to select the most appropriate therapeutic intervention prior to surgery. This study evaluates the applicability on fine‐needle aspirations (FNAs) of the novel Kiupel grading system, based on number of mitoses, multinucleated cells, bizarre nuclei and presence of karyomegaly. Fifty consecutive cases with pre‐operative cytological diagnosis were included. In cytological specimens, approximately 1000 cells were evaluated, and the histological grade was assessed on the corresponding resected specimens. On cytology, the above parameters were significantly different between histologically low‐grade and high‐grade tumours (P < 0.001). The cytograding correctly predicted the histological grade in 47 cases (accuracy, 94%; sensitivity, 84.6%; specificity, 97.3%). Two high‐grade MCTs (4%) were not detected on cytology. The cytograding can provide helpful insights to assist clinical decisions in most cases. However, the risk of underestimation in a minority of patients represents a limit to the overall utility of the technique.  相似文献   

Acute‐phase proteins as diagnostic markers in horses with colic          下载免费PDF全文

Tina H. Pihl DVM  PhD  Elrien Scheepers BVSc  MSc  Macarena Sanz DVM  MS  DACVIM  Amelia Goddard BVSc  MMed Vet  Nils Toft Cand Sci  PhD  Mads Kjelgaard‐Hansen DVM  PhD  Pia H. Andersen DVM  PhD  DVSc  Stine Jacobsen DVM  PhD 《Journal of Veterinary Emergency and Critical Care》2016,26(5):664-674

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