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| 1 | One-step generation of myostatin gene knockout sheep via the CRISPR/Cas9 system显示文摘The CRISPR/Cas(clustered regularly interspaced short palindromic repeats/CRISPR-associated)adaptive immune system,which was discovered in bacteria and archaea,can specifically degrade invasive viral and plasmid DNA by base pairing between crRNAs(CRISPR RNAs)and the target DNA[1,2].Recently,the Streptococcus pyogenes type II CRISPR system was shown to be able to perform efficient targeted gene disruption by employing three fundamental components:(1)Cas9 endonuclease. | Hongbing HAN Yonghe MA Tao WANG Ling LIAN Xiuzhi TIAN Rui HU Shoulong DENG Kongpan LI Feng WANG Ning LI Guoshi LIU Yaofeng ZHAO Zhengxing LIAN | 2014 | Frontiers of Agricultural Science and Engineering2014，1，1： | 8 |
| 2 | Resveratrol compares with melatonin in improving in vitro porcine oocyte maturation under heat stress显示文摘Background: Resveratrol, an important phyto-antioxidant commonly found in grapes, mulberry, and other plants,has a variety of functions including anti-aging, anti-cancer and anti-inflammatory activities. In the current study, we investigated the beneficial effects of resveratrol on in vitro porcine oocyte maturation under heat stress(HS). The effect of resveratrol, melatonin and their combination on alleviating HS was compared according to the maturation rate of oocytes and the development competence of embryos after parthenogenetic activation(PA).Results: Supplementation with resveratrol(2.0 μmol/L) not only improved the nuclear maturation but also raised the blastocyst rate of porcine embryos' PA from oocytes that underwent HS by increasing their glutathione(GSH)level, reducing reactive oxygen species(ROS) and up-regulating the expression of Sirtuin 1(SIRT1). It was also found that melatonin(10^(-7)mol/L) and the combination of resveratrol(2.0 μmol/L) plus melatonin(10^(-7)mol/L) exhibited more potent effects than resveratrol alone regarding their protective activities on oocyte maturation under HS.Conclusions: This study compared the efficiencies of resveratrol, melatonin and their combination for protecting porcine oocytes from heat stress. The mechanisms are attributed to the fact that each treatment may have different ability to regulate the synthesis of steroid hormones and the expression of mature related genes. | Yu Li Jing Wang Zhenzhen Zhang Jinyun Yi Changjiu He Feng Wang Xiuzhi Tian Minghui Yang Yukun Song Pingli He Guoshi Liu | 2017 | Journal of Animal Science and Biotechnology2017，8，1： | 7 |
| 3 | Over-expression of Toll-like receptor 2 up-regulates heme oxygenase-1 expression and decreases oxidative injury in dairy goats显示文摘Background: Mastitis, an infection caused by Gram-positive bacteria, produces udder inflammation and oxidative injury in milk-producing mammals. Toll-like receptor 2(TLR2) is important for host recognition of invading Grampositive microbes. Over-expression of TLR2 in transgenic dairy goats is a useful model for studying various aspects of infection with Gram-positive bacteria, in vivo.Methods: We over-expressed TLR2 in transgenic dairy goats. Pam3CSK4, a component of Gram-positive bacteria,triggered the TLR2 signal pathway by stimulating the monocytes-macrophages from the TLR2-positive transgenic goats, and induced over-expression of activator protein-1(AP-1), phosphatidylinositol 3-kinase(PI3K) and transcription factor nuclear factor kappa B(NF-κB) and inflammation factors downstream of the signal pathway.Results: Compared with wild-type controls, measurements of various oxidative stress-related molecules showed that TLR2, when over-expressed in transgenic goat monocytes-macrophages, resulted in weak lipid damage, high level expression of anti-oxidative stress proteins, and significantly increased m RNA levels of transcription factor NF-E2-related factor-2(Nrf2) and the downstream gene, heme oxygenase-1(HO-1). When Pam3CSK4 was used to stimulate ear tissue in vivo the HO-1 protein of the transgenic goats had a relatively high expression level.Conclusions: The results indicate that the oxidative injury in goats over-expressing TLR2 was reduced following Pam3CSK4 stimulation. The underlying mechanism for this reduction was increased expression of the anti-oxidation gene HO-1 by activation of the Nrf2 signal pathway. | Shoulong Deng Kun Yu Wuqi Jiang Yan Li Sutian Wang Zhuo Deng Yuchang Yao Baolu Zhang Guoshi Liu Yixun Liu Zhengxing Lian | 2017 | Journal of Animal Science and Biotechnology2017，8，2： | 2 |
| 4 | Scanning pattern of diffusion tensor tractography and an analysis of the morphology and function of spinal nerve roots显示文摘Radiculopathy,commonly induced by intervertebral disk bulging or protrusion,is presently diagnosed in accordance with clinical symptoms because there is no objective quantitative diagnostic criterion.Diffusion tensor magnetic resonance imaging and diffusion tensor tractography revealed the characterization of anisotropic diffusion and displayed the anatomic form of nerve root fibers This study included 18 cases with intervertebral disc degeneration-induced unilateral radiculopathy Magnetic resonance diffusion tensor imaging was creatively used to reveal the scanning pattern o fiber tracking of the spinal nerve root.A scoring system of nerve root morphology was used to quantitatively assess nerve root morphology and functional alteration after intervertebral disc degeneration.Results showed that after fiber tracking,compared with unaffected nerve root,fiber bundles gathered together and interrupted at the affected side.No significant alteration was detected in the number of fiber bundles,but the cross-sectional area of nerve root fibers was reduced These results suggest that diffusion tensor magnetic resonance imaging-based tractography can be used to quantitatively evaluate nerve root function according to the area and morphology of fiber bundles of nerve roots. | Xin Tian Huaijun Liu Zuojun Geng Hua Yang Guoshi Wang Jiping Yang Chunxia Wang Cuining Li Ying Li | 2013 | Neural Regeneration Research2013，8，33： | 2 |
| 5 | Efficient production of pronuclear embryos in breeding and nonbreeding season for generating transgenic sheep overexpressing TLR4显示文摘Background: Brucella is a zoonotic Gram-negative pathogen that causes abortion and infertility in ruminants and humans. TLR4 is the receptor for LPS which can recognize Brucella and initiate antigen-presenting cell activities that affect both innate and adaptive immunity. Consequently, transgenic sheep over-expressing TLR4 are an suitable model to investigate the effects of TLR4 on preventing Brucellosis. In this study, we generated transgenic sheep overexpressing TLR4 and aimed to evaluate the effects of different seasons(breeding and non-breeding season) on superovulation and the imported exogenous gene on growth.Results: In total of 43 donor ewes and 166 recipient ewes in breeding season, 37 donor ewes and 144 recipient ewes in non-breeding season were selected for super-ovulation and injected embryo transfer to generate transgenic sheep.Our results indicated the no. of embryos recovered of donors and the rate of pronuclear embryos did not show any significant difference between breeding and non-breeding seasons(P > 0.05). The positive rate of exogenous TLR4 tested were 21.21 % and 22.58 % in breeding and non-breeding season by Southern blot. The expression level of TLR4 in the transgenic sheep was 1.5 times higher than in the non-transgenic group(P < 0.05). The lambs overexpressing TLR4 had similar growth performance with non-transgenic lambs, and the blood physiological parameters of transgenic and non-transgenic were both in the normal range and did not show any difference.Conclusions: Here we establish an efficient platform for the production of transgenic sheep by the microinjection of pronuclear embryos during the whole year. The over-expression of TLR4 had no adverse effect on the growth of the sheep. | Yan Li Di Lian Shoulong Deng Xiaosheng Zhang Jinlong Zhang Wenting Li Hai Bai Zhixian Wang Hongping Wu Juncai Fu Hongbing Han Jianzhong Feng Guoshi Liu Ling Lian Zhengxing Lian | 2017 | Journal of Animal Science and Biotechnology2017，8，1： | 2 |
| 6 | Neuroprotective effect of shenfu injection after intra-arterial thrombolysis显示文摘BACKGROUND: Within 3 hours after ischemic stroke, thrombolysis can improve prognosis of patients; however, cerebral edema and cerebral hemorrhage induced by vascular recanalization at an early phase can influence on therapeutic effect of thrombolysis; therefore, thrombolysis is combined with neuroprotective agent for recently clinical treatment. OBJECTIVE: To evaluate the neuroprotective effect of shenfu injection on patients with ischemic stroke after arterial thrombolysis and compare with the controls. DESIGN: Case-controlled study. SETTING: Department of Medical Image, Second Hospital of Hebei Medical University. PARTICIPANTS: A total of 45 patients with acute stroke were selected from Emergency Department or Neurological Department of the Second Hospital of Hebei Medical University from August 2005 to May 2006. All patients were met the diagnosis criteria published by the Fourth National Stroke Conference and had middle cerebral arterial embolism with angiography. Attacked course was less than 6 hours. Patients did not have a history of stroke. Scores of the National Institutes of Health Stroke Scale (NIHSS) were not less than 4 points and patients were aged from 18 to 80 years. All subjects consented of the whole therapeutic process. METHODS: ① Intermittent perfusion was used to treat intra-arterial thrombolysis. Thirty patients with vascular recanalization or partially vascular recanalization (18 males and 12 females) were randomly divided into treatment group and control group with 15 in each group. There was no significant difference of baseline data between the two groups. After recanalization, patients in treatment group were given 20 mL shenfu injection, which consisted of ginseng saponin and aconitine (Ya’an Sanjiu Pharmaceutical Company, batch number: 041013, 10 mL/ampoule) and slowly injected into vein for longer than 5 minutes. And then, patients were intravenously dripped with 50 mL shenfu injection and 50 g/L 250 mL glucose solution once a day for 2 courses. Fourteen days were regarded as a course. Patients in control group were treated with the same volume of glucose solution (50 g/L), and other processes in this group were as the same as those in treatment group. ② NIHSS scores before thrombolysis and at 24 hours after thrombolysis: Patients were scored as the follows: level of consciousness, staring, sight, facial paralysis, upper limb movement, lower limb movement, incoordination, sense, language, dysarthria and neglect. Total scores ranged from 0 to 42, and the more the scores were, the more obvious the symptom was. ③ Patients were checked with head CT examination at 24 hours after thrombolysis. ④ Modified Rankin scores at 3 months after thrombolysis: Total scores ranged from 0 to 6. 0: symptomless; 6: death; 0-2: well; 3-6: poor. ⑤ Adverse events and side effects were checked after thrombolysis. MAIN OUTCOME MEASURES: ① Comparison of NIHSS scores between the two groups before and after thrombolysis; ② comparison of modified Rankin scores at 3 months after thrombolysis; ③ comparison of adverse events and side effects. RESULTS: Among 45 patients with ischemic stroke, 30 ones with vascular recanalization were involved in the final analysis. ① Comparison of NIHSS scores before and after thrombolysis: After thrombolysis, NIHSS scores were 5 in treatment group and 8 in control group, which were lower than those before thrombolysis (12, 13; U =2.054, 1.652; P < 0.05). At 24 hours after thrombolysis, NIHSS score was 5 in treatment group, which was lower than that in control group (8, U =2.157, P < 0.05). ② Comparison of modified Rankin scores at 3 months after thrombolysis: Rankin scores which were regarded as well accounted for 93% (14/15) in treatment group, which was higher than those in control group (53%, 8/15, P < 0.05). ③ Comparison of adverse events and side effects: Patients in treatment group did not have cerebral hemorrhage and whole-body hemorrhage with head CT examination; meanwhile, all patients were living at 24 hours after thrombolysis. Among 2 patients with cerebral hemorrhage, 1 case were dead in control group with head CT examination at 24 hours after thrombolysis. CONCLUSION: Intra-arterial thrombolysis combined with shenfu injection plays a neuroprotective role in ischemic stroke; additionally, it is safe for patients. | Huaijun Liu Xiuchuan Jia Jiping Yang Zengpin Liu Guoshi Wang Linfang Li | 2006 | Neural Regeneration Research2006，1，5： | 1 |
| 7 | Whole-genome methylation analysis reveals epigenetic variation between wild-type and nontransgenic cloned,ASMT transgenic cloned dairy goats generated by the somatic cell nuclear transfer显示文摘Background:SCNT(somatic cell nuclear transfer)is of great significance to biological research and also to the livestock breeding.However,the survival rate of the SCNT cloned animals is relatively low compared to other transgenic methods.This indicates the potential epigenetic variations between them.DNA methylation is a key marker of mammalian epigenetics and its alterations will lead to phenotypic differences.In this study,ASMT(acetylserotonin-Omethyltransferase)ovarian overexpression transgenic goat was produced by using SCNT.To investigate whether there are epigenetic differences between cloned and WT(wild type)goats,WGBS(whole-genome bisulfite sequencing)was used to measure the whole-genome methylation of these animals.Results:It is observed that the different m Cp G sites are mainly present in the intergenic and intronic regions between cloned and WT animals,and their CG-type methylation sites are strongly correlated.DMR(differentially methylated region)lengths are located around 1000 bp,mainly distributed in the exonic,intergenic and intronic functional domains.A total of 56 and 36 DMGs(differentially methylated genes)were identified by GO and KEGG databases,respectively.Functional annotation showed that DMGs were enriched in biological-process,cellularcomponent,molecular-function and other signaling pathways.A total of 10 identical genes related to growth and development were identified in GO and KEGG databases.Conclusion:The differences in methylation genes among the tested animals have been identified.A total of 10 DMGs associated with growth and development were identified between cloned and WT animals.The results indicate that the differential patterns of DNA methylation between the cloned and WT goats are probably caused by the SCNT.These novel observations will help us to further identify the unveiled mechanisms of somatic cell cloning technology,particularly in goats. | Hao Wu Wendi Zhou Haijun Liu Xudai Cui Wenkui Ma Haixin Wu Guangdong Li Likai Wang Jinlong Zhang Xiaosheng Zhang Pengyun Ji Zhengxing Lian Guoshi Liu | 2023 | Journal of Animal Science and Biotechnology2023，14，1： | 0 |
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