Ovine bones are the major by-products after slaughtered. The present study was conducted to extract and characterize acid soluble collagens （ASC） and pepsin soluble collagens （PSC） from ovine bones （Ujumuqin shee...Ovine bones are the major by-products after slaughtered. The present study was conducted to extract and characterize acid soluble collagens （ASC） and pepsin soluble collagens （PSC） from ovine bones （Ujumuqin sheep）. Ovine bones collagen were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE） and liquid chromatography-tan- dem mass spectrometry （LC-MS/MS） as type I collagen. The results of Fourier transform infrared （FTIR） spectra analysis testified the existence of triple superhelical structure in both ASC and PSC, showing pepsin did not disrupt the triple helical structure of ovine bones collagen. Glycine, accounting for one-third of total amino acids, was the major amino acid for ovine bones collagen. Higher imino acid content was responsible for higher thermal denaturation temperature of ovine bones collagen compared to fish collagens. The isoelectric point of ASC was lower than PSC due to the higher content of acidic amino acids. Therefore, this study provides the potential reference for collagen extraction and application of ovine bones by-procduct.展开更多
Over 550000 people in the U.S.require hemodialysis for management of end stage renal disease(ESRD).When anatomy restricts fistula creation,arteriovenous grafts(AVG)are implanted.AVGs have poor primary patency and high...Over 550000 people in the U.S.require hemodialysis for management of end stage renal disease(ESRD).When anatomy restricts fistula creation,arteriovenous grafts(AVG)are implanted.AVGs have poor primary patency and high risk of infection,highlighting a need for better alternatives.Previous AVG large animal models were limited by high complication rates and short-term follow-up.This study investigates the safety and durability of an ovine bilateral carotid-jugular AVG model.Eight female sheep underwent bilateral carotid-jugular AVG implantation via a single longitudinal incision overlying the trachea.Ringed PTFE grafts were anastomosed in a“lazy-S”configuration to provide the laxity needed to prevent avulsion and minimize kinking with neck movement.Post-operatively,sheep were evaluated daily to monitor for complications.Duplex ultrasonography of the grafts was performed at regular intervals out to 6 months to evaluate patency.At 6 months,angiography and duplex was performed followed by explant for gross and histologic analysis.Technical success was achieved in 16 of 16(100%)graft implants.No major complications,including stroke,anastomotic disruption,infection,wound breakdown,or death occurred.Primary patency of control PTFE grafts was 75%at 6 months,paralleling reported rates in humans.Bilateral carotid-jugular AVG implantation in sheep is a safe and durable model for self-controlled long-term evaluation of AVG conduit technology.展开更多
Tuberculosis is Zoonotic disease and infected animals continue to spread the disease to other animals and humans by excreting the germs in milk, meat, feces, and respiratory droplets, this study used bPPD (Bovine puri...Tuberculosis is Zoonotic disease and infected animals continue to spread the disease to other animals and humans by excreting the germs in milk, meat, feces, and respiratory droplets, this study used bPPD (Bovine purified protein derivative) to observe TST (Tuberculin skin test) reactivity in the Sheep population in the Sadar and Parbatipur Upazilas of Dinajpur District. The research conducted from January to June of 2018. There is no history for sheep tuberculosis in the study area. Overall, 5 percent (7/140) of 140 sheep were positive, 15% (21/140) were doubtful, and 80% (112/140) were negative based on sex, age, breed, and body condition score, while lactation stage and reproduction status prevalence was 6.4 percent, 14.9 percent, and 78.7% positive, doubtful, and negative, respectively. Our findings suggest the presence of a skin test reaction response when employing the bPPD reagent, but this needs to be validated by pathological and serological techniques. Nevertheless, my preliminary investigation suggests that abattoirs must implement effective control systems and raise public awareness.展开更多
What is special about the sky bridge near Qingjing Farm?Qingjing Farm in Nantou is a popular place to visit.Every year,it has a Sheep Run Festival!Thousands of visitors go each year to cheer on their favorite sheep an...What is special about the sky bridge near Qingjing Farm?Qingjing Farm in Nantou is a popular place to visit.Every year,it has a Sheep Run Festival!Thousands of visitors go each year to cheer on their favorite sheep and take pictures as they run down the mountain roads.People can enjoy many performances,games and activities during the festival,too.展开更多
Background The deposition of intramuscular fat(IMF)in livestock can enhance the flavor and tenderness of meat products,significantly increasing consumer satisfaction.To achieve this industrial trait,this study investi...Background The deposition of intramuscular fat(IMF)in livestock can enhance the flavor and tenderness of meat products,significantly increasing consumer satisfaction.To achieve this industrial trait,this study investigated the regulatory effects of 20 dietary nutrients on sheep IMF deposition using a 3D organoid culture model.Results Key nutrients enhancing angiogenesis,adipocyte differentiation,and lipid accumulation were identified through assessments of capillary sprouts development,mRNA expression,and Oil Red O staining.Vitamins C(VC),E(VE),and K1(VK1),guanidinoacetic acid(GAA),leucine(Leu),lysine(Lys),methionine(Met),N-carbamylglutamate(NCG),tryptophan(Trp),α-linolenic acid(ALA),linoleic acid(LA),cis-9,trans-11 conjugated linoleic acid(c9,t11-CLA),acetic acid(HAc),and sodium acetate(NaAc)stimulated while vitamins B9(VB9),D(VD),K2(VK2),taurine(Tau),and sodium butyrate(NaBu)inhibited angiogenesis(P<0.05).Furthermore,VC,VE,VK1,VK2,GAA,Leu,NCG,Trp,ALA,LA,and HAc enhanced adipocyte differentiation,with VE,VK1,GAA,Leu,LA,and HAc additionally elevating lipid accumulation(P<0.05).Conclusions Various nutrients play distinct regulatory roles in angiogenesis,adipocyte differentiation,and lipid accumulation.These findings provide a roadmap for further optimizing the production of marbled meat through nutritional intervention in actual livestock breeding production.展开更多
In order to improve the meat production performance of local sheep varieties in Gansu Province, Dorset was introduced to crossbreed with the local sheep varieties, including Tan sheep, Small Tail Han sheep and Mongoli...In order to improve the meat production performance of local sheep varieties in Gansu Province, Dorset was introduced to crossbreed with the local sheep varieties, including Tan sheep, Small Tail Han sheep and Mongolia sheep. The offspring of different crossbreeding combinations were sampled randomly at different growth stages, and their growth and development traits were measured so as to screen out the best crossbreeding mode. The results showed that under the same crossbreeding mode, the growth rate of F3 was higher than that of F2, and of F2 was higher than that of F1. Among the F3 population, the growth rates of Dorset ×Han and Dorset × Mongolia hybrids were higher. Compared with those of Dorset ×Tan F3 hybrids, the body weights of male and female Dorset × Han and Dorset ×Mongolia F3 hybrids were increased by 5.59%, 4.40%, 5.93% and 3.76%, respectively. Among the F2 population, the growth rates of Dorset × Han and Dorset ×Mongolia hybrids were also higher. The body weights of male and female Dorset ×Han and Dorset × Mongolia F2 hybrids were higher than those of Dorset × Tan ×Han F2 hybrids by 5.99%, 3.67%, 9.80% and 5.00%, respectively. In the F1 population, the growth rates of Dorset × Han and Dorset × Mongolia hybrids were higher.Compared with those of Tan × Han F1 hybrids, the body weights of male and female Dorset × Han and Dorset × Mongolia F1 hybrids were increased by 11.32%,5.22%, 7.60% and 7.20%, respectively. Therefore, in the feeding area of Small Tai Han sheep, Mongolia sheep and Tan sheep, Dorset was the best sire for producing hybrid lambs. The economic benefit of crossbred offspring was obvious.展开更多
Objective] This study aimed to clone ovine activin receptor type llB （Ac-tRIIB） gene, construct the prokaryotic expression vector and express the target gene in vitro, thus providing basis for further function verif...Objective] This study aimed to clone ovine activin receptor type llB （Ac-tRIIB） gene, construct the prokaryotic expression vector and express the target gene in vitro, thus providing basis for further function verification. [Method] The template cDNA which was reversely transcribed from total RNA of sheep liver tissue, was subjected to polymerase chain reaction （PCR） using specific primers of ActRIIB. The ful-length cDNA of ovine ActRIIB was obtained by pMD18-T cloning and sequencing for bioinformatics analysis. Ovine ActRIIB encoding sequence was subcloned into prokaryotic expression vector pET41a with restriction sites BamHl/Notl, and then transformed into BL21 （DE3）. The induced products by lPTG were analyzed with SDS-PAGE and Western Blot. [Result] The amplified ful-length cDNA of ovine Ac-tRllB gene was 1 564 bp in length （Genbank accession number： JX422071.1） with an open reading frame of 1 539 bp, encoding 512 amino acides. Ovine ActRllB shared the highest homology （99.6%） with bovine ActRllB. ActRllB had highly ho-mologous C-terminal domains and belonged to the TGFβ family. After prokaryotic expression, an approximately 92 kD His-tagged ActRllB recombinant protein was obtained, which was consistent with the excepted result. [Conclusion] Cloning and successful expression of ovine ActRIIB laid solid foundation for further investigation of its biological function.展开更多
Objective Cryptosporidium spp. are prevalent globally and sheep are an important zoonotic reservoir. Little data regarding the rates of Cryptosporidium infections in ovines in China are available. This study assessed ...Objective Cryptosporidium spp. are prevalent globally and sheep are an important zoonotic reservoir. Little data regarding the rates of Cryptosporidium infections in ovines in China are available. This study assessed the prevalence of Cryptosporidium spp. in pre-weaned ovines from Aba Tibetan and Qiang Autonomous Prefecture in the Sichuan province of China. Methods A total of 213 fecal samples were collected from pre-weaned ovines and were examined microscopically （following modified acid fast staining）. In addition, 18S rRNA genetic sequences were amplified from fecal samples by nested PCR and phylogenetically analyzed. Results The prevalence of Cryptosporidium in the collected samples was at 14.6% （31/213） and four isolates identified by PCR belonged to the Cryptosporidium cervine genotype （Cryptosporidium ubiquiturn） demonstrating that this species was the primary sheep species found in sheep in China. Conclusion The present study suggested that the high incidence of Cryptosporidium in sheep poses a significant public health threat and that surveillance practices must be established to prevent zoonotic disease of humans.展开更多
In order to improve the meat performance of local sheep in Gansu Province,Dorset and Borderdale were introduced to crossbreed with local sheep which were Tan sheep,Small-tail Han sheep and Mongolia sheep. The offsprin...In order to improve the meat performance of local sheep in Gansu Province,Dorset and Borderdale were introduced to crossbreed with local sheep which were Tan sheep,Small-tail Han sheep and Mongolia sheep. The offspring under different crossbreeding combinations were sampled randomly at the different growing stage to measure their growth traits so as to select optimize the crossbreeding mode. The results indicated that,for the same crossbreeding mode,the growth rate of progeny was in order F3】 F2】 F1; for the F3 progeny,the combinations Dorset- Borderdale- Small tail Han sheep and Dorset- Borderdale- Mongolia sheep gave a higher growth rate,with a body weight of 1. 57%,3. 17%,8. 23%,1. 15% higher in male and female individuals than the counterparts of Dorset and Tan sheep and Small tail Han sheep; for the F2 progeny,the combinations Dorset- Borderdale- Small tail Han sheep and Dorset- Borderdale- Mongolia sheep also gave a higher growth rate,with a body weight of 2. 15%,4. 53%,9. 21% and 2. 75% higher in male and female individuals than the counterparts of Dorset and Tan sheep and Small tail Han sheep; for the F1 progeny,the combination Borderdale and Small tail Han sheep assumed a higher growth rate,with a body weight of 3. 23%,6. 07%,7. 42% and 8. 66% higher in male and female individuals than the counterparts of Borderdale- Mongolia sheep and Tan sheep- Small tail Han sheep,respectively. Therefore,in the Small-tail Han sheep and Mongolia sheep producing regions,the F2 or F3progeny bred by using Dorset or Borderdale sheep as male parent to cross with local breeds,or the hybrid lambs of Small-tail Han sheep and Borderdale sheep as highly qualified commodity,would produce significant economic benefit. Moreover,the novel breeds obtained by crossing were the valuable genetic resource for breeding meat sheep.展开更多
This study aimed to examine changes in phosphorylation of sarcoplasmic and myofibrillar proteins from longissimus lumborum,semitendinosus,and psoas major muscles during postmortem ageing for 5 d.These sarcoplasmic and...This study aimed to examine changes in phosphorylation of sarcoplasmic and myofibrillar proteins from longissimus lumborum,semitendinosus,and psoas major muscles during postmortem ageing for 5 d.These sarcoplasmic and myofibrillar proteins were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and stained with phosphorous and protein specific stains.Myofibril fragmentation index,pH,the content of lactic acid and the relative activity of μ-calpain in three ovine muscles were measured.These results showed that the relative phosphorylation level of sarcoplasmic and myofibrillar proteins of psoas major muscle were lower compared with longissimus lumborum and semitendinosus muscles(P<0.05).The pH of psoas major muscle was the lowest at 0.5 h postmortem,and the highest after 12 h postmortem(P<0.05).In addition,the relative activity of μ-calpain was higher within 5 d postmortem and myofibril fragmentation index was higher after 1 d postmortem in psoas major muscle than those of longissimus lumborum and semitendinosus muscles(P<0.05).The sarcoplasmic protein phosphorylation may regulate the rate of pH decline to influence the μ-calpain activity and then proteolysis of proteins consequently.This study gives a new perspective of the mechanism of postmortem meat tenderization.展开更多
Phosphorylation post-translational modification plays an important role in postmortem muscle quality traits. Adenosine triphosphate(ATP) is an energy source and a key substrate of phosphorylation which provides the ph...Phosphorylation post-translational modification plays an important role in postmortem muscle quality traits. Adenosine triphosphate(ATP) is an energy source and a key substrate of phosphorylation which provides the phosphatase groups to proteins in the presence of protein kinases. However, in postmortem muscle, the effects of ATP content on phosphorylation are poorly studied. The study investigated the effect of ATP on protein phosphorylation and degradation in postmortem ovine muscle. The ground muscle with/without additional ATP were treated/control groups and stored at 25 and 4℃, respectively. The ATP content led to different changes of p H value between the ATP-treated and control groups. The phosphorylation level of myofibrillar proteins was higher(P<0.05) in ATP-treated group compared to the control group at both temperatures, which suggested that ATP played a vital role in postmortem protein phosphorylation. A slower degradation rate of μ-calpain, desmin and troponin T was observed in the ATP-treated group which showed that there was a negative relationship between ATP level and the degradation of proteins. These observations clearly highlighted the role of ATP on the development of meat quality by regulating the phosphorylation and degradation of myofibrillar proteins in postmortem ovine muscle.展开更多
Background:Heavy metal cadmium(Cd)is a widespread environmental contaminant with a potential toxicity that might negatively affect female reproduction and fertility.It has been reported that Cd exposure impaired the q...Background:Heavy metal cadmium(Cd)is a widespread environmental contaminant with a potential toxicity that might negatively affect female reproduction and fertility.It has been reported that Cd exposure impaired the quality of oocytes and led to a defective maturation and fertilization,through oxidative stress induction.Resveratrol(Res)is a natural polyphenol with strong antioxidant properties that exhibited protective role in preventing oocyte redox homeostasis disruption and quality decline.Here,we explored whether the addition of Res to in vitro maturation(IVM)medium might act as a protection against Cd-induced toxicity on ovine oocyte maturation and fertilization.Firstly,we evaluated the effect of supplementing IVM medium with two different Res concentrations(1and 2μmol/L)on nuclear maturation and fertilization of oocytes matured under CdCl2(2μmol/L)exposure.Therefore,the concentration of 1μmol/L Res was selected to analyse the effects of this compound on intracellular ROS levels,mitochondrial(mt)distribution and activity,chromatin configuration,cytoskeleton morphology,cortical granules(CGs)distribution and mRNA expression of genes associated with cellular response to oxidative stress(i.e.SIRT1,SOD 1,GPX1,GSR,CAT)in Cd-exposed in vitro matured oocytes.Results:We found that 1μmol/L Res restored the reduced oocyte meiotic competence induced by Cd exposure as well as,Res sustained oocyte ability to be normally fertilized and decreased polyspermic fertilization at both tested concentrations.Moreover,we demonstrated that 1μmol/L Res mitigated Cd-induced alterations of oocyte cytoplasmic maturation by reducing reactive oxygen species(ROS)accumulation,preventing mt dysfunction,maintaining the correct meiotic spindle and cortical F-actin assembly and the normal cortical granule distribution as well as up-regulating SIRT1,SOD1 and GPX1 genes.Conclusions:Taken together,our findings highlighted the beneficial influence exerted by Res in preventing Cdinduced disturbance of nuclear and cytoplasmic maturation and subsequent fertilization in ovine oocytes.Res treatment may help to establish defence strategies counteracting Cd-induced toxicity on the female gamete.展开更多
为筛选出策勒黑羊种质资源繁殖和抗逆性状分子标记和相关基因,本研究以100只无亲缘关系的策勒黑羊为研究对象,基于Illumina ovine SNP 50 K bead chip进行基因分型;用Plink1.90对基因组数据进行质量控制;对有效SNPs计算哈代-温伯格值,...为筛选出策勒黑羊种质资源繁殖和抗逆性状分子标记和相关基因,本研究以100只无亲缘关系的策勒黑羊为研究对象,基于Illumina ovine SNP 50 K bead chip进行基因分型;用Plink1.90对基因组数据进行质量控制;对有效SNPs计算哈代-温伯格值,选择从小到大排列后前1%的分子标记,参考绵羊基因组(Oar_v4.0)进行注释,对注释到的基因进行GO和KEGG分析。结果表明:1)共获得480个SNPs,417个候选基因,其中,繁殖性状基因13个、抗逆性状相关基因30个;2)繁殖性状相关的基因有SOX9、GTF2A1、GNAQ等,抗逆性相关的基因有TMEM154、ZNF70、CREB3L2等。综上,本研究解析了策勒黑羊繁殖和抗逆性状基因的遗传规律,特别是对抗逆性相关候选基因研究,发现策勒黑羊群体携带抗肺炎相关的分子标记,为策勒黑羊保种和改良提供了分子水平的参考。展开更多
This study was conducted to investigate the drug sensitivity of Escherichia coli isolated from sheep, providing data reference for clinical medication. Fifty-four samples were collected from a certain sheep farm in Ya...This study was conducted to investigate the drug sensitivity of Escherichia coli isolated from sheep, providing data reference for clinical medication. Fifty-four samples were collected from a certain sheep farm in Yangling, Shaanxi Province for E. coli isolation and identification, and K-B disk diffusion method was used to carry out the dug sensitivity test of 40 E. coli to the eight antimicrobials of spectinomycin, amikacin, tobramycin, enrofloxacin, nalidixic acid, levofloxacin, salafloxacin and ciprofloxacin. The results showed that there were 40 positive E. coli strains and the isolation rate was 74.7%. The resistance rates of the 40 E. coli to spectinomycin, amikacin,tobramycin and nalidixic acid were 85%, 95%, 97.5% and 87.5%, respectively, while the resistance rate to enrofloxacin, levofloxacin, salafloxacin and ciprofloxacin were relatively low and the resistance rates were 35%, 20%, 25% and 17.5%, respectively. The isolates showed different levels of drug resistance to eight antimicrobials and the highest resistance rate was 97.5%. It reminds that the sheep farm should strictly control the dosage according to the antimicrobial sensitive data in clinic.展开更多
Objective:To assess the effect of cryopreservation on stemness and proliferation potential of sheep spermatogonial stem cells(SSCs)in vitro.Methods:Sheep testicular cells were isolated and putative SSCs were enriched ...Objective:To assess the effect of cryopreservation on stemness and proliferation potential of sheep spermatogonial stem cells(SSCs)in vitro.Methods:Sheep testicular cells were isolated and putative SSCs were enriched by the laminin-based differential plating method.Putative SSCs were co-cultured with the Sertoli cell feeder prepared by the Datura Stramonium Agglutinin(DSA-lectin)-based method.The cultured putative SSCs were cryopreserved in Dulbecco's Modified Eagle Medium-10%fetal bovine serum mixture(DMEM-10%FBS)media containing 10%dimethyl sulfoxide(DMSO)alone or 10%DMSO plus 200 mM trehalose.Cryopreserved putative SSCs were evaluated for their proliferation potential using in vitro culture and stemness by immunocytochemistry.Finally,the transfection ability of cryopreserved putative SSCs was analyzed.Results:We isolated 91%viable testicular cells from sheep testes.The majority of the laminin enriched cells expressed the SSC related marker,ITGA6.Co-culture of sheep putative SSCs with Sertoli cell feeder resulted in the generation of stable colonies,and the expression of SSC marker was maintained after several passages.A significantly higher number of viable putative SSCs was recovered from SSCs cryopreserved in media containing 10%DMSO and 200 mM trehalose compared to 10%DMSO alone(P<0.01).Cryopreserved putative SSCs formed colonies and showed SSC marker expression similar to the non-cryopreserved putative SSCs.The appearance of green fluorescent colonies over the Sertoli cell feeder indicated that cryopreserved sheep SSCs were successfully transfected.Conclusions:Cryopreserved putative SSCs can retain their stemness,colony forming ability,and transfection efficiency in vitro.Our research may help in the effective preservation of germplasm and the generation of transgenic ovine species.展开更多
Recently the use of biologic materials as dura mater repair patches has been increasing. The purpose of this study is to assess the basis for efficacy and safety of using a novel fish derived acellular dermis (Kerecis...Recently the use of biologic materials as dura mater repair patches has been increasing. The purpose of this study is to assess the basis for efficacy and safety of using a novel fish derived acellular dermis (Kerecis Omega3 DuraTM). In an ovine model a craniotomy under general anaesthesia was performed. A defect was produced in the dural covering of approximately 1 × 2 cm and closed with an onlay technique, with Kerecis Omega3 Dura. The bone defect was covered with the bony flap and the overlying tissues closed in layers. At 2, 5, 8 and 11 weeks the sheep underwent MRI scanning followed by euthanasia, necropsy and histological assessment. MRI images taken at 2, 5, 8 and 11 weeks showed initially moderate inflammatory response, which diminished over time, and at 11 weeks no evidence of inflammation existed. There was evidence of cerebrospinal fluid leakage at no time point. Necropsy revealed some adhesions at 5 and 8 weeks, in particular at 5 weeks, but at 11 weeks there were no adhesions found. From 2 - 11 weeks, there was evidence of initially an inflammatory reaction followed by neodura formation at the defect site through cellular ingrowth and remodeling of the acellular fish skin. Histology showed a histiocytic foreign body reaction initially that subsided over time. As early as 8 weeks there was evidence of neodura formation and by 11 weeks there was a minimal inflammatory response with an intact neodura formed. In this pilot study the Kerecis Omega3 Dura patches performed in a safe and efficacious manner. This new material needs to be fully assessed and compared with other products that are currently on the market in a larger scale animal study.展开更多
The study investigated the effects of epidermal growth factor（EGF） and insulin-like growth factor 1 （ IGF-I）, alone or together, on the in vitro maturation and cleavage of ovine oocytes, aimed to optimize the in v...The study investigated the effects of epidermal growth factor（EGF） and insulin-like growth factor 1 （ IGF-I）, alone or together, on the in vitro maturation and cleavage of ovine oocytes, aimed to optimize the in vitro maturation conditions for ovine oocytes. The results showed that the maturation and cleavage rates were 71.2% and 45.5% respectively when the medium was supplemented with 50 ng/mL EGF alone, which was significantly higher than other EGF supplemented groups （0, 10, 20, 30, and 40 ng/mL） （P 〈0.05）. The highest maturation and cleavage rates were 72.9% and 45. 7% when the EGF concentration reached 100 ng/mL. The maturation and cleavage rates were 70. 7% and 58. 5% with 40 ng/mL IGF-I supplemented, which were significantly higher than other treatments （0,10,20,60,80, and 100 ng/mL） （P 〈0.05）. The lowest maturation and cleavage rates were 38.8% and 20.0% when the IGF-I concentration reached 100 ng/mL （ P 〈 0.05 ） When 50 ng/mL EGF and 40 ng/mL IGF-I were used concomitantly,the maturation and cleavage rates were 85. 6% and 61. 0% respectively, which were significantly higher than the treatments with EGF or IGF-I alone （ P 〈 0.05 ）.展开更多
Objective To investigate the development and characterizations of the hepatocytes isolated from fetal ovine and to determine the effect of hypoxia on their growth and metabolism.Methods Fresh hepatocytes were isolated...Objective To investigate the development and characterizations of the hepatocytes isolated from fetal ovine and to determine the effect of hypoxia on their growth and metabolism.Methods Fresh hepatocytes were isolated from the liver of fetal ovine at late gestation, cultured in specific media, and exposed to normoxia(21% O2) or hypoxia(2% O2).The cellular characteristics and population purity were identified by immunocytochemistry and flow cytometry(FCM).The effects of hypoxia on cell cycle and apoptosis of the hepatocytes were evaluated by FCM, whereas the cellular ultrastructure changes were examined with a transmission electron microscope.Results The cell purity of hepatocytes was over 95%.Under hypoxia exposure, the hepatocytes showed a gradual increase in proportion at the S phase and in proliferative index, followed with a compatible increase in apoptosis and progressively decreased cell viability.Additionally, the organelles of the hepatocytes demonstrated dramatic changes, including swelling of mitochondria, disorder in cristae arrangement, expansion of endoplasmic reticulum, and a large number of circular lipid droplets emerging in the cytoplasm.Conclusion Fetal ovine hepatocytes could be primarily cultured in a short-term culture system with a high purity of over 95% and with their preserved original characteristics.Hypoxia could induce changes in ultrastructural and inhibit the proliferation of cultured fetal ovine hepatocytes through apoptotic mechanisms.展开更多
This study is aimed to investigate the effect of luteinizing hormone （LH） on maturation and fertilization in vitro of ovine oocytes. The ovine oocytes were cultured in vitro in medium with or without LH, and then ch...This study is aimed to investigate the effect of luteinizing hormone （LH） on maturation and fertilization in vitro of ovine oocytes. The ovine oocytes were cultured in vitro in medium with or without LH, and then checked by confocal laser scanning microscope to observe the distribution of cortical granules stained with FITC-LCA during different maturation periods. Similarly, some in vitro matured oocytes were also fertilized in vitro for analysis of their developmental potentiality further. After being cultured in vitro for 4 h, there were significant differences about the rate of germinal vesicle break down （GVBD） between the treatment （with LH） and the control groups （without any hormones） （36.76% vs 18%, P〈0.05）. Further, there were also significant differences of the cleavage and blastocyst rates between these two groups （67.15% vs 42.37%, 21.9% vs 12.71%, P〈0.05, respectively）. The distribution of cortical granules appeared to spread from the edges to the central site of sheep oocytes following their delaying durations of maturation in vitro. It can be concluded that LH may play a role to delay the occurence of GVBD, prolong the maturation duration of cytoplasm, and enhance the nuclear and cytoplasm synchronization of ovine oocytes matured in vitro and finally improve their in vitro developmental potentiality.展开更多
Background： Maternal over and restricted nutrition has negative consequences on the muscle of offspring by reducing muscle fiber number and altering regulators of muscle growth. To determine if over and restricted ma...Background： Maternal over and restricted nutrition has negative consequences on the muscle of offspring by reducing muscle fiber number and altering regulators of muscle growth. To determine if over and restricted maternal nutrition affected muscle growth and gene fed 60%, 100% or 140% of National Research Council and protein expression in offspring, 36 pregnant ewes were requirements from d 31 ＋ 1,3 of gestation until parturition. Lambs from control-fed （CON）, restricted-fed （RES） or over-fed （OVER） ewes were necropsied within 1 d of birth （n = 18） or maintained on a control diet for 3 mo （n = ] 5）. Semitendinosus muscle was collected for immunohistochemistry, and protein and gene expression analysis. Results： Compared with CON, muscle fiber cross-sectional area （CSA） increased in RES （58%） and OVER （47%） lambs at 1 d of age （P〈 0.01）; however at 3 mo, CSA decreased 15% and 17% compared with CON, respectively （P 〈 0.01）. Compared with CON, muscle lipid content was increased in OVER （212.4%） and RES （92.5%） at d 1 （P 〈 0.0001）. Muscle lipid content was increased 36.1% in OVER and decreased 23.6% in RES compared with CON at 3 mo （P 〈 0.0001）. At d 1, myostatin mRNA abundance in whole muscle tended to be greater in OVER （P = 0.07） than CON. Follistatin mRNA abundance increased in OVER （P = 0.04） and tended to increase in RES （P = 0.06） compared with CON at d 1. However, there was no difference in myostatin or follistatin protein expression （P 〉 0.3）. Phosphorylated Akt （ser473） was increased in RES at 3 mo compared with CON （P = 0.006）. Conclusions： In conclusion, maternal over and restricted nutrient intake alters muscle lipid content and growth of offspring, possibly through altered gene and protein expression.展开更多
基金funded by the emarked fund for China Agriculture Research System (CARS-39)the National Agricultural Science and Technology Innovation Program
文摘Ovine bones are the major by-products after slaughtered. The present study was conducted to extract and characterize acid soluble collagens （ASC） and pepsin soluble collagens （PSC） from ovine bones （Ujumuqin sheep）. Ovine bones collagen were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE） and liquid chromatography-tan- dem mass spectrometry （LC-MS/MS） as type I collagen. The results of Fourier transform infrared （FTIR） spectra analysis testified the existence of triple superhelical structure in both ASC and PSC, showing pepsin did not disrupt the triple helical structure of ovine bones collagen. Glycine, accounting for one-third of total amino acids, was the major amino acid for ovine bones collagen. Higher imino acid content was responsible for higher thermal denaturation temperature of ovine bones collagen compared to fish collagens. The isoelectric point of ASC was lower than PSC due to the higher content of acidic amino acids. Therefore, this study provides the potential reference for collagen extraction and application of ovine bones by-procduct.
基金California Institute for Regenerative Medicine(CIRM),Grant/Award Number:TRAN3-13332。
文摘Over 550000 people in the U.S.require hemodialysis for management of end stage renal disease(ESRD).When anatomy restricts fistula creation,arteriovenous grafts(AVG)are implanted.AVGs have poor primary patency and high risk of infection,highlighting a need for better alternatives.Previous AVG large animal models were limited by high complication rates and short-term follow-up.This study investigates the safety and durability of an ovine bilateral carotid-jugular AVG model.Eight female sheep underwent bilateral carotid-jugular AVG implantation via a single longitudinal incision overlying the trachea.Ringed PTFE grafts were anastomosed in a“lazy-S”configuration to provide the laxity needed to prevent avulsion and minimize kinking with neck movement.Post-operatively,sheep were evaluated daily to monitor for complications.Duplex ultrasonography of the grafts was performed at regular intervals out to 6 months to evaluate patency.At 6 months,angiography and duplex was performed followed by explant for gross and histologic analysis.Technical success was achieved in 16 of 16(100%)graft implants.No major complications,including stroke,anastomotic disruption,infection,wound breakdown,or death occurred.Primary patency of control PTFE grafts was 75%at 6 months,paralleling reported rates in humans.Bilateral carotid-jugular AVG implantation in sheep is a safe and durable model for self-controlled long-term evaluation of AVG conduit technology.
文摘Tuberculosis is Zoonotic disease and infected animals continue to spread the disease to other animals and humans by excreting the germs in milk, meat, feces, and respiratory droplets, this study used bPPD (Bovine purified protein derivative) to observe TST (Tuberculin skin test) reactivity in the Sheep population in the Sadar and Parbatipur Upazilas of Dinajpur District. The research conducted from January to June of 2018. There is no history for sheep tuberculosis in the study area. Overall, 5 percent (7/140) of 140 sheep were positive, 15% (21/140) were doubtful, and 80% (112/140) were negative based on sex, age, breed, and body condition score, while lactation stage and reproduction status prevalence was 6.4 percent, 14.9 percent, and 78.7% positive, doubtful, and negative, respectively. Our findings suggest the presence of a skin test reaction response when employing the bPPD reagent, but this needs to be validated by pathological and serological techniques. Nevertheless, my preliminary investigation suggests that abattoirs must implement effective control systems and raise public awareness.
文摘What is special about the sky bridge near Qingjing Farm?Qingjing Farm in Nantou is a popular place to visit.Every year,it has a Sheep Run Festival!Thousands of visitors go each year to cheer on their favorite sheep and take pictures as they run down the mountain roads.People can enjoy many performances,games and activities during the festival,too.
基金supported by the Key Research and Development Program-Key Projects(2021YFD1200900,2023YFD1301302)the grants from the National Natural Science Foundation of China(32272892,32472886)+2 种基金the Bayannaoer Research Institute Young Scientist Project(2024BYNECAU003)the Chinese Universities Scientific Fund(2025TC101)the 2115 Talent Development Program of China Agricultural University。
文摘Background The deposition of intramuscular fat(IMF)in livestock can enhance the flavor and tenderness of meat products,significantly increasing consumer satisfaction.To achieve this industrial trait,this study investigated the regulatory effects of 20 dietary nutrients on sheep IMF deposition using a 3D organoid culture model.Results Key nutrients enhancing angiogenesis,adipocyte differentiation,and lipid accumulation were identified through assessments of capillary sprouts development,mRNA expression,and Oil Red O staining.Vitamins C(VC),E(VE),and K1(VK1),guanidinoacetic acid(GAA),leucine(Leu),lysine(Lys),methionine(Met),N-carbamylglutamate(NCG),tryptophan(Trp),α-linolenic acid(ALA),linoleic acid(LA),cis-9,trans-11 conjugated linoleic acid(c9,t11-CLA),acetic acid(HAc),and sodium acetate(NaAc)stimulated while vitamins B9(VB9),D(VD),K2(VK2),taurine(Tau),and sodium butyrate(NaBu)inhibited angiogenesis(P<0.05).Furthermore,VC,VE,VK1,VK2,GAA,Leu,NCG,Trp,ALA,LA,and HAc enhanced adipocyte differentiation,with VE,VK1,GAA,Leu,LA,and HAc additionally elevating lipid accumulation(P<0.05).Conclusions Various nutrients play distinct regulatory roles in angiogenesis,adipocyte differentiation,and lipid accumulation.These findings provide a roadmap for further optimizing the production of marbled meat through nutritional intervention in actual livestock breeding production.
文摘In order to improve the meat production performance of local sheep varieties in Gansu Province, Dorset was introduced to crossbreed with the local sheep varieties, including Tan sheep, Small Tail Han sheep and Mongolia sheep. The offspring of different crossbreeding combinations were sampled randomly at different growth stages, and their growth and development traits were measured so as to screen out the best crossbreeding mode. The results showed that under the same crossbreeding mode, the growth rate of F3 was higher than that of F2, and of F2 was higher than that of F1. Among the F3 population, the growth rates of Dorset ×Han and Dorset × Mongolia hybrids were higher. Compared with those of Dorset ×Tan F3 hybrids, the body weights of male and female Dorset × Han and Dorset ×Mongolia F3 hybrids were increased by 5.59%, 4.40%, 5.93% and 3.76%, respectively. Among the F2 population, the growth rates of Dorset × Han and Dorset ×Mongolia hybrids were also higher. The body weights of male and female Dorset ×Han and Dorset × Mongolia F2 hybrids were higher than those of Dorset × Tan ×Han F2 hybrids by 5.99%, 3.67%, 9.80% and 5.00%, respectively. In the F1 population, the growth rates of Dorset × Han and Dorset × Mongolia hybrids were higher.Compared with those of Tan × Han F1 hybrids, the body weights of male and female Dorset × Han and Dorset × Mongolia F1 hybrids were increased by 11.32%,5.22%, 7.60% and 7.20%, respectively. Therefore, in the feeding area of Small Tai Han sheep, Mongolia sheep and Tan sheep, Dorset was the best sire for producing hybrid lambs. The economic benefit of crossbred offspring was obvious.
基金Supported by Natural Science Foundation of Xinjiang Uygur Autonomous Region(2012211B54)~~
文摘Objective] This study aimed to clone ovine activin receptor type llB （Ac-tRIIB） gene, construct the prokaryotic expression vector and express the target gene in vitro, thus providing basis for further function verification. [Method] The template cDNA which was reversely transcribed from total RNA of sheep liver tissue, was subjected to polymerase chain reaction （PCR） using specific primers of ActRIIB. The ful-length cDNA of ovine ActRIIB was obtained by pMD18-T cloning and sequencing for bioinformatics analysis. Ovine ActRIIB encoding sequence was subcloned into prokaryotic expression vector pET41a with restriction sites BamHl/Notl, and then transformed into BL21 （DE3）. The induced products by lPTG were analyzed with SDS-PAGE and Western Blot. [Result] The amplified ful-length cDNA of ovine Ac-tRllB gene was 1 564 bp in length （Genbank accession number： JX422071.1） with an open reading frame of 1 539 bp, encoding 512 amino acides. Ovine ActRllB shared the highest homology （99.6%） with bovine ActRllB. ActRllB had highly ho-mologous C-terminal domains and belonged to the TGFβ family. After prokaryotic expression, an approximately 92 kD His-tagged ActRllB recombinant protein was obtained, which was consistent with the excepted result. [Conclusion] Cloning and successful expression of ovine ActRIIB laid solid foundation for further investigation of its biological function.
基金supported by grants from the Chinese Special Program for Scientific Research of Public Health (200802012)Chinese National Key Program for Infectious Diseases of China (2009ZX10004‐201, and 2008ZX10004‐002)
文摘Objective Cryptosporidium spp. are prevalent globally and sheep are an important zoonotic reservoir. Little data regarding the rates of Cryptosporidium infections in ovines in China are available. This study assessed the prevalence of Cryptosporidium spp. in pre-weaned ovines from Aba Tibetan and Qiang Autonomous Prefecture in the Sichuan province of China. Methods A total of 213 fecal samples were collected from pre-weaned ovines and were examined microscopically （following modified acid fast staining）. In addition, 18S rRNA genetic sequences were amplified from fecal samples by nested PCR and phylogenetically analyzed. Results The prevalence of Cryptosporidium in the collected samples was at 14.6% （31/213） and four isolates identified by PCR belonged to the Cryptosporidium cervine genotype （Cryptosporidium ubiquiturn） demonstrating that this species was the primary sheep species found in sheep in China. Conclusion The present study suggested that the high incidence of Cryptosporidium in sheep poses a significant public health threat and that surveillance practices must be established to prevent zoonotic disease of humans.
基金Supported by Science&Technology Pillar Program of Gansu Province(1104NKCA083)
文摘In order to improve the meat performance of local sheep in Gansu Province,Dorset and Borderdale were introduced to crossbreed with local sheep which were Tan sheep,Small-tail Han sheep and Mongolia sheep. The offspring under different crossbreeding combinations were sampled randomly at the different growing stage to measure their growth traits so as to select optimize the crossbreeding mode. The results indicated that,for the same crossbreeding mode,the growth rate of progeny was in order F3】 F2】 F1; for the F3 progeny,the combinations Dorset- Borderdale- Small tail Han sheep and Dorset- Borderdale- Mongolia sheep gave a higher growth rate,with a body weight of 1. 57%,3. 17%,8. 23%,1. 15% higher in male and female individuals than the counterparts of Dorset and Tan sheep and Small tail Han sheep; for the F2 progeny,the combinations Dorset- Borderdale- Small tail Han sheep and Dorset- Borderdale- Mongolia sheep also gave a higher growth rate,with a body weight of 2. 15%,4. 53%,9. 21% and 2. 75% higher in male and female individuals than the counterparts of Dorset and Tan sheep and Small tail Han sheep; for the F1 progeny,the combination Borderdale and Small tail Han sheep assumed a higher growth rate,with a body weight of 3. 23%,6. 07%,7. 42% and 8. 66% higher in male and female individuals than the counterparts of Borderdale- Mongolia sheep and Tan sheep- Small tail Han sheep,respectively. Therefore,in the Small-tail Han sheep and Mongolia sheep producing regions,the F2 or F3progeny bred by using Dorset or Borderdale sheep as male parent to cross with local breeds,or the hybrid lambs of Small-tail Han sheep and Borderdale sheep as highly qualified commodity,would produce significant economic benefit. Moreover,the novel breeds obtained by crossing were the valuable genetic resource for breeding meat sheep.
基金funded by the National Agricultural Science and Technology Innovation Program in China
文摘This study aimed to examine changes in phosphorylation of sarcoplasmic and myofibrillar proteins from longissimus lumborum,semitendinosus,and psoas major muscles during postmortem ageing for 5 d.These sarcoplasmic and myofibrillar proteins were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and stained with phosphorous and protein specific stains.Myofibril fragmentation index,pH,the content of lactic acid and the relative activity of μ-calpain in three ovine muscles were measured.These results showed that the relative phosphorylation level of sarcoplasmic and myofibrillar proteins of psoas major muscle were lower compared with longissimus lumborum and semitendinosus muscles(P<0.05).The pH of psoas major muscle was the lowest at 0.5 h postmortem,and the highest after 12 h postmortem(P<0.05).In addition,the relative activity of μ-calpain was higher within 5 d postmortem and myofibril fragmentation index was higher after 1 d postmortem in psoas major muscle than those of longissimus lumborum and semitendinosus muscles(P<0.05).The sarcoplasmic protein phosphorylation may regulate the rate of pH decline to influence the μ-calpain activity and then proteolysis of proteins consequently.This study gives a new perspective of the mechanism of postmortem meat tenderization.
基金financial support from the National Natural Science Foundation of China (31771995)the earmarked fund for China Agriculture Research System (CARS-38)the Agricultural Science and Technology Innovation Program, Chinese Academy of Agricultural Sciences (CAAS-ASTIP-IFST)。
文摘Phosphorylation post-translational modification plays an important role in postmortem muscle quality traits. Adenosine triphosphate(ATP) is an energy source and a key substrate of phosphorylation which provides the phosphatase groups to proteins in the presence of protein kinases. However, in postmortem muscle, the effects of ATP content on phosphorylation are poorly studied. The study investigated the effect of ATP on protein phosphorylation and degradation in postmortem ovine muscle. The ground muscle with/without additional ATP were treated/control groups and stored at 25 and 4℃, respectively. The ATP content led to different changes of p H value between the ATP-treated and control groups. The phosphorylation level of myofibrillar proteins was higher(P<0.05) in ATP-treated group compared to the control group at both temperatures, which suggested that ATP played a vital role in postmortem protein phosphorylation. A slower degradation rate of μ-calpain, desmin and troponin T was observed in the ATP-treated group which showed that there was a negative relationship between ATP level and the degradation of proteins. These observations clearly highlighted the role of ATP on the development of meat quality by regulating the phosphorylation and degradation of myofibrillar proteins in postmortem ovine muscle.
基金funded by Fondazione Banco di Sardegna,FDS 2016(CUP J86C18000780005 and J86C18000810005)。
文摘Background:Heavy metal cadmium(Cd)is a widespread environmental contaminant with a potential toxicity that might negatively affect female reproduction and fertility.It has been reported that Cd exposure impaired the quality of oocytes and led to a defective maturation and fertilization,through oxidative stress induction.Resveratrol(Res)is a natural polyphenol with strong antioxidant properties that exhibited protective role in preventing oocyte redox homeostasis disruption and quality decline.Here,we explored whether the addition of Res to in vitro maturation(IVM)medium might act as a protection against Cd-induced toxicity on ovine oocyte maturation and fertilization.Firstly,we evaluated the effect of supplementing IVM medium with two different Res concentrations(1and 2μmol/L)on nuclear maturation and fertilization of oocytes matured under CdCl2(2μmol/L)exposure.Therefore,the concentration of 1μmol/L Res was selected to analyse the effects of this compound on intracellular ROS levels,mitochondrial(mt)distribution and activity,chromatin configuration,cytoskeleton morphology,cortical granules(CGs)distribution and mRNA expression of genes associated with cellular response to oxidative stress(i.e.SIRT1,SOD 1,GPX1,GSR,CAT)in Cd-exposed in vitro matured oocytes.Results:We found that 1μmol/L Res restored the reduced oocyte meiotic competence induced by Cd exposure as well as,Res sustained oocyte ability to be normally fertilized and decreased polyspermic fertilization at both tested concentrations.Moreover,we demonstrated that 1μmol/L Res mitigated Cd-induced alterations of oocyte cytoplasmic maturation by reducing reactive oxygen species(ROS)accumulation,preventing mt dysfunction,maintaining the correct meiotic spindle and cortical F-actin assembly and the normal cortical granule distribution as well as up-regulating SIRT1,SOD1 and GPX1 genes.Conclusions:Taken together,our findings highlighted the beneficial influence exerted by Res in preventing Cdinduced disturbance of nuclear and cytoplasmic maturation and subsequent fertilization in ovine oocytes.Res treatment may help to establish defence strategies counteracting Cd-induced toxicity on the female gamete.
文摘为筛选出策勒黑羊种质资源繁殖和抗逆性状分子标记和相关基因,本研究以100只无亲缘关系的策勒黑羊为研究对象,基于Illumina ovine SNP 50 K bead chip进行基因分型;用Plink1.90对基因组数据进行质量控制;对有效SNPs计算哈代-温伯格值,选择从小到大排列后前1%的分子标记,参考绵羊基因组(Oar_v4.0)进行注释,对注释到的基因进行GO和KEGG分析。结果表明:1)共获得480个SNPs,417个候选基因,其中,繁殖性状基因13个、抗逆性状相关基因30个;2)繁殖性状相关的基因有SOX9、GTF2A1、GNAQ等,抗逆性相关的基因有TMEM154、ZNF70、CREB3L2等。综上,本研究解析了策勒黑羊繁殖和抗逆性状基因的遗传规律,特别是对抗逆性相关候选基因研究,发现策勒黑羊群体携带抗肺炎相关的分子标记,为策勒黑羊保种和改良提供了分子水平的参考。
基金Supported by the Fundamental Research Funds for the Central Universities:the Young Teachers Fund of Southwest Minzu University(2017NZYQN38)Innovative Scientific Research Project for Postgraduates of Southwest Minzu University(CX2018SZ42)
文摘This study was conducted to investigate the drug sensitivity of Escherichia coli isolated from sheep, providing data reference for clinical medication. Fifty-four samples were collected from a certain sheep farm in Yangling, Shaanxi Province for E. coli isolation and identification, and K-B disk diffusion method was used to carry out the dug sensitivity test of 40 E. coli to the eight antimicrobials of spectinomycin, amikacin, tobramycin, enrofloxacin, nalidixic acid, levofloxacin, salafloxacin and ciprofloxacin. The results showed that there were 40 positive E. coli strains and the isolation rate was 74.7%. The resistance rates of the 40 E. coli to spectinomycin, amikacin,tobramycin and nalidixic acid were 85%, 95%, 97.5% and 87.5%, respectively, while the resistance rate to enrofloxacin, levofloxacin, salafloxacin and ciprofloxacin were relatively low and the resistance rates were 35%, 20%, 25% and 17.5%, respectively. The isolates showed different levels of drug resistance to eight antimicrobials and the highest resistance rate was 97.5%. It reminds that the sheep farm should strictly control the dosage according to the antimicrobial sensitive data in clinic.
文摘Objective:To assess the effect of cryopreservation on stemness and proliferation potential of sheep spermatogonial stem cells(SSCs)in vitro.Methods:Sheep testicular cells were isolated and putative SSCs were enriched by the laminin-based differential plating method.Putative SSCs were co-cultured with the Sertoli cell feeder prepared by the Datura Stramonium Agglutinin(DSA-lectin)-based method.The cultured putative SSCs were cryopreserved in Dulbecco's Modified Eagle Medium-10%fetal bovine serum mixture(DMEM-10%FBS)media containing 10%dimethyl sulfoxide(DMSO)alone or 10%DMSO plus 200 mM trehalose.Cryopreserved putative SSCs were evaluated for their proliferation potential using in vitro culture and stemness by immunocytochemistry.Finally,the transfection ability of cryopreserved putative SSCs was analyzed.Results:We isolated 91%viable testicular cells from sheep testes.The majority of the laminin enriched cells expressed the SSC related marker,ITGA6.Co-culture of sheep putative SSCs with Sertoli cell feeder resulted in the generation of stable colonies,and the expression of SSC marker was maintained after several passages.A significantly higher number of viable putative SSCs was recovered from SSCs cryopreserved in media containing 10%DMSO and 200 mM trehalose compared to 10%DMSO alone(P<0.01).Cryopreserved putative SSCs formed colonies and showed SSC marker expression similar to the non-cryopreserved putative SSCs.The appearance of green fluorescent colonies over the Sertoli cell feeder indicated that cryopreserved sheep SSCs were successfully transfected.Conclusions:Cryopreserved putative SSCs can retain their stemness,colony forming ability,and transfection efficiency in vitro.Our research may help in the effective preservation of germplasm and the generation of transgenic ovine species.
文摘Recently the use of biologic materials as dura mater repair patches has been increasing. The purpose of this study is to assess the basis for efficacy and safety of using a novel fish derived acellular dermis (Kerecis Omega3 DuraTM). In an ovine model a craniotomy under general anaesthesia was performed. A defect was produced in the dural covering of approximately 1 × 2 cm and closed with an onlay technique, with Kerecis Omega3 Dura. The bone defect was covered with the bony flap and the overlying tissues closed in layers. At 2, 5, 8 and 11 weeks the sheep underwent MRI scanning followed by euthanasia, necropsy and histological assessment. MRI images taken at 2, 5, 8 and 11 weeks showed initially moderate inflammatory response, which diminished over time, and at 11 weeks no evidence of inflammation existed. There was evidence of cerebrospinal fluid leakage at no time point. Necropsy revealed some adhesions at 5 and 8 weeks, in particular at 5 weeks, but at 11 weeks there were no adhesions found. From 2 - 11 weeks, there was evidence of initially an inflammatory reaction followed by neodura formation at the defect site through cellular ingrowth and remodeling of the acellular fish skin. Histology showed a histiocytic foreign body reaction initially that subsided over time. As early as 8 weeks there was evidence of neodura formation and by 11 weeks there was a minimal inflammatory response with an intact neodura formed. In this pilot study the Kerecis Omega3 Dura patches performed in a safe and efficacious manner. This new material needs to be fully assessed and compared with other products that are currently on the market in a larger scale animal study.
文摘The study investigated the effects of epidermal growth factor（EGF） and insulin-like growth factor 1 （ IGF-I）, alone or together, on the in vitro maturation and cleavage of ovine oocytes, aimed to optimize the in vitro maturation conditions for ovine oocytes. The results showed that the maturation and cleavage rates were 71.2% and 45.5% respectively when the medium was supplemented with 50 ng/mL EGF alone, which was significantly higher than other EGF supplemented groups （0, 10, 20, 30, and 40 ng/mL） （P 〈0.05）. The highest maturation and cleavage rates were 72.9% and 45. 7% when the EGF concentration reached 100 ng/mL. The maturation and cleavage rates were 70. 7% and 58. 5% with 40 ng/mL IGF-I supplemented, which were significantly higher than other treatments （0,10,20,60,80, and 100 ng/mL） （P 〈0.05）. The lowest maturation and cleavage rates were 38.8% and 20.0% when the IGF-I concentration reached 100 ng/mL （ P 〈 0.05 ） When 50 ng/mL EGF and 40 ng/mL IGF-I were used concomitantly,the maturation and cleavage rates were 85. 6% and 61. 0% respectively, which were significantly higher than the treatments with EGF or IGF-I alone （ P 〈 0.05 ）.
基金funded by National Natural Science Foundation [81370719 and 81671535]Jiangsu Science Foundation [BE2015642]+3 种基金Jiangsu Key Discipline of Human Assisted Reproduction Medicine Foundation [FXK201749]Jiangsu Provincial Medical Talent of the Project of Invigorating Healthcare through Science,Technology and Education [ZDRCA2016044]and Chinese Medical Association Clinical Medicine Research Reproductive Medicine [17020270696]The Priority Academic Program Development of the Jiangsu Higher Education Institutes(PAPD)
文摘Objective To investigate the development and characterizations of the hepatocytes isolated from fetal ovine and to determine the effect of hypoxia on their growth and metabolism.Methods Fresh hepatocytes were isolated from the liver of fetal ovine at late gestation, cultured in specific media, and exposed to normoxia(21% O2) or hypoxia(2% O2).The cellular characteristics and population purity were identified by immunocytochemistry and flow cytometry(FCM).The effects of hypoxia on cell cycle and apoptosis of the hepatocytes were evaluated by FCM, whereas the cellular ultrastructure changes were examined with a transmission electron microscope.Results The cell purity of hepatocytes was over 95%.Under hypoxia exposure, the hepatocytes showed a gradual increase in proportion at the S phase and in proliferative index, followed with a compatible increase in apoptosis and progressively decreased cell viability.Additionally, the organelles of the hepatocytes demonstrated dramatic changes, including swelling of mitochondria, disorder in cristae arrangement, expansion of endoplasmic reticulum, and a large number of circular lipid droplets emerging in the cytoplasm.Conclusion Fetal ovine hepatocytes could be primarily cultured in a short-term culture system with a high purity of over 95% and with their preserved original characteristics.Hypoxia could induce changes in ultrastructural and inhibit the proliferation of cultured fetal ovine hepatocytes through apoptotic mechanisms.
基金supported in part by the grants fromthe National Natural Science Foundation of China(30871836) the Key Fund of Natural Science from Beijing Municipal Government, China (Type B,KZ200510020013)
文摘This study is aimed to investigate the effect of luteinizing hormone （LH） on maturation and fertilization in vitro of ovine oocytes. The ovine oocytes were cultured in vitro in medium with or without LH, and then checked by confocal laser scanning microscope to observe the distribution of cortical granules stained with FITC-LCA during different maturation periods. Similarly, some in vitro matured oocytes were also fertilized in vitro for analysis of their developmental potentiality further. After being cultured in vitro for 4 h, there were significant differences about the rate of germinal vesicle break down （GVBD） between the treatment （with LH） and the control groups （without any hormones） （36.76% vs 18%, P〈0.05）. Further, there were also significant differences of the cleavage and blastocyst rates between these two groups （67.15% vs 42.37%, 21.9% vs 12.71%, P〈0.05, respectively）. The distribution of cortical granules appeared to spread from the edges to the central site of sheep oocytes following their delaying durations of maturation in vitro. It can be concluded that LH may play a role to delay the occurence of GVBD, prolong the maturation duration of cytoplasm, and enhance the nuclear and cytoplasm synchronization of ovine oocytes matured in vitro and finally improve their in vitro developmental potentiality.
文摘Background： Maternal over and restricted nutrition has negative consequences on the muscle of offspring by reducing muscle fiber number and altering regulators of muscle growth. To determine if over and restricted maternal nutrition affected muscle growth and gene fed 60%, 100% or 140% of National Research Council and protein expression in offspring, 36 pregnant ewes were requirements from d 31 ＋ 1,3 of gestation until parturition. Lambs from control-fed （CON）, restricted-fed （RES） or over-fed （OVER） ewes were necropsied within 1 d of birth （n = 18） or maintained on a control diet for 3 mo （n = ] 5）. Semitendinosus muscle was collected for immunohistochemistry, and protein and gene expression analysis. Results： Compared with CON, muscle fiber cross-sectional area （CSA） increased in RES （58%） and OVER （47%） lambs at 1 d of age （P〈 0.01）; however at 3 mo, CSA decreased 15% and 17% compared with CON, respectively （P 〈 0.01）. Compared with CON, muscle lipid content was increased in OVER （212.4%） and RES （92.5%） at d 1 （P 〈 0.0001）. Muscle lipid content was increased 36.1% in OVER and decreased 23.6% in RES compared with CON at 3 mo （P 〈 0.0001）. At d 1, myostatin mRNA abundance in whole muscle tended to be greater in OVER （P = 0.07） than CON. Follistatin mRNA abundance increased in OVER （P = 0.04） and tended to increase in RES （P = 0.06） compared with CON at d 1. However, there was no difference in myostatin or follistatin protein expression （P 〉 0.3）. Phosphorylated Akt （ser473） was increased in RES at 3 mo compared with CON （P = 0.006）. Conclusions： In conclusion, maternal over and restricted nutrient intake alters muscle lipid content and growth of offspring, possibly through altered gene and protein expression.
