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Transfection and expression of exogenous gene in laying hens oviduct in vitro and in vivo

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Journal of Zhejiang university-Science B(Biomedicine & Biotechnology) 2005年第2期6卷 137-141页

作者：高波孙怀昌宋成义王志跃陈芹宋红芹 School of veterinary medicine Yangzhou university Yangzhou 225009 China School of veterinary mediciney

To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region. The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyle-neimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis. The results showed that expression of pEGFP-Nl was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the β-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.

关键词： Chicken ovalbumin gene regulatory regions Oviduct-specific expression vector Oviduct epithelium In vivo expression

Primary oral and nasal transmissible venereal tumor in a mix-breed dog

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Asian Pacific Journal of Tropical Biomedicine 2016年第5期6卷 443-445页

作者： Mahdieh Rezaei Shahrzad Azizi Shima Shahheidaripour Sara Rostami Department of Clinical Science Faculty of Veterinary MedicineShahid Bahonar University Department of Pathobiology Faculty of Veterinary MedicineShahid Bahonar University Faculty of veterinary medicine Shahid Bahonar University

Transmissible venereal tumor(TVT) is a coitally transmitted tumor of dogs with widespread distribution. The present study describes the occurrence of the primary oral and nasal TVT in a 10-year-old, female, mix-breed dog. The case was presented with a history of anorexia, inability to swallow and dyspnea. Clinical examinations revealed the emaciation, muzzle deformity due to the presence of a friable, fleshy, cauliflower-like mass in the oral cavity and submandibular lymphadenopathy. TVT was diagnosed based on histopathological findings. The dog was discharged with therapeutic intervention with vincristine. Unfortunately, the case died before readmission because of the progressive worsening of the general condition. Our findings highlight the need for considering TVT for the differential diagnosis of the extragenital masses in dogs.

关键词： Transmissible venereal tumor Primary Oral Nasal Dog

The Finding and Phylogenetic Evolution Analysis of Bovine Piroplasms in the Rasǒn Area of North Korea

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Journal of Integrative Agriculture 2013年第10期12卷 1847-1854页

作者： JIA Li-jun ZHANG Shou-fa CAO Shi-nuo QIAN Nian-chao YU Long-zheng XUAN Xue-nan Laboratory of veterinary Microbiology Department of Veterinary MedicineYanbian University National Research Center for Protozoan Diseases Obihiro University of Agriculture and Veterinary MedicineInada-choObihiroHokkaido080-8555Japan

The objective of this study was to investigate the epidemiology of bovine Piroplasms infections in the Rasǒn area of North *** survey was carried out by light microscopic examination of Giemsa-stained blood smears,PCR,and phylogenetic evolution analysis of 128 blood samples collected from the Rasǒn *** results showed that the infection rates of the small and large parasites were about 2.5 and 1.5% on average,respectively,in all Theileria sergenti and Babesia ovatapositive blood smears by microscopic examination of blood *** detection rate of *** Giemsa-stained smears was 43.75%,while that with PCR was 67.97%.The detection rate of *** Giemsa-stained smears was 49.21%,while that with PCR was 71.88%.The sequence and phylogenetic analysis of DNA showed 98.84% homology between the 18S rRNA gene sequences of *** isolates from North Korean and that of Yanbian state from China,indicating the closest genetic relationship between both of ***,100% homology was shown between the 18S rRNA gene sequence of *** isolates from North Korea and the published sequence AY081192 of GenBank,indicating the closest genetic relationship between both of *** survey confirmed that Ras n is the endemic area of *** and *** in North Korea.

关键词： Rasǒn area of North Korea bovine Piroplasms phylogenetic trees

Efficacy of voriconazole on leishmaniasis by Leishmania major: An in vitro and in vivo study

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Asian Pacific Journal of Tropical medicine 2018年第10期11卷 562-569页

作者： Ahmad Oryan Somayeh Bahrami Effat Bemani Department of Pathology School of Veterinary Medicine Shiraz University Department of Parasitology Faculty of Veterinary Medicine Shahid Chamran University of Ahvaz

Objective: To appraise the activity of voriconazole against Leishmania major(L. major) in vitro and its effectiveness on wound regeneration in cutaneous leishmaniasis in BALB/c mice Methods: The IC_(50) of voriconazole against promastigotes and intra-macrophage amastigotes of L. major was investigated in vitro. The in vivo study was performed by treating the L. major infected BALB/c mice. When the wounds appeared in the base of tail, treatment was started by administration of 30 mg/kg voriconazole for 28 consecutive days orally. Results: The IC_(50) of voriconazole against promastigotes and intra-macrophage amastigotes were 0.74 and 0.89μM, respectively. Voriconazole decreased lipid peroxidation and IL-6 level. Histopathologica findings indicated accelerated healing in the voriconazole treated group compared to other groups. Conclusions: Our results demonstrate that voriconazole can be an option in treating the cutaneous leishmaniasis by L. major.

关键词： Leishmania major Wound healing Azoles Voriconazole Histopathology

Cardiopulmonary,Biochemical and Haematological Effects of the Tiletamine/Zolazepam-Xylazine-Tramadol Combination to Provide Anaesthesia in Miniature Pigs

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Journal of Integrative Agriculture 2012年第8期11卷 1340-1346页

作者： LU De-zhang FAN Hong-gang JIANG Sheng ZHANG Luan-song MA Kun YU Shi-ming TAN Li-juan WANG Hong-bin Department of veterinary Surgery College of Veterinary Medicine Northeast Agricultural University Harbin 150030 IER. China Department of veterinary Surgery College of Veterinary Medicine Northwest Agricultrue and Forestry University Yangling 712100 P..R China

The objective of this study was to investigate the suitability of tiletamine/zolazepam-xylazine-tramadol combination for miniature pigs. Fourteen Chinese experimental miniature pigs subjected to this study received 3.5 mg tiletamine/zolazepam kg-1 bw, 1.32 mg xylazine kg-1 bw and 1.8 mg tramadol kg-1 bw intramuscularly, as a mixture of the drugs. Cardiopulmonary, biochemical and haematological parameters were recorded before drug administration and after anaesthesia. The combination of the compounds resulted in anaesthesia lasting about 87 min and a satisfactory immobilization for handling. Cardiopulmonary parameters were changed after administration, but there were within biologically acceptable limits. Biochemical and haematological values decreased after drug administration, however, they returned to the baseline at 24 h. At the doses described, tiletamine/zolazepam-xylazine-tramadol combination produced good immobilization in miniature pigs with minimal changes over time in cardiopulmonary, biochemical and haematological parameters.

关键词： cardiopulmonary tiletamine/zolazepam-xylaxine-tramadol combination anaesthesia miniature pigs

Changes in expression of inhibitory substances in the intramural neurons of the stomach following streptozotocininduced diabetes in the pig

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World Journal of Gastroenterology 2017年第33期23卷 6088-6099页

作者： Michal Bulc Katarzyna Palus Lukasz Zielonka Magdalena Gajecka Jaroslaw Całka Department of Clinical Physiology Faculty of veterinary medicine University of Warmia and Mazury Department of veterinary Prevention and Feed Hygiene Faculty of Veterinary MedicineUniversity of Warmia and Mazury in Olsztyn

AIM Influence of chronic hyperglycemia on chemical coding of enteric neurons in stomach using pig as a model for human diabetic *** Ten pigs were divided into two groups: diabetic(D group,n = 5) and control(C group,n = 5). Pigs constituting the experimental group were given streptozotocin(150 mg/kg). Animals were euthanized six weeks after the induction of diabetes. The samples of stomach were collected from animals of both groups. The cryostat sections were processed for double immunofluorescence staining using primary antisera directed towards pan-neuronal marker(Hu C/D) proteins and/or neuronal isoform of nitric oxide synthase(n NOS),vasoactive intestinal peptide(VIP) and galanin(GAL). RESULTS In the control group in the myenteric ganglia(MG) of the corpus we have noted 22.28% ± 1.19% of n NOS positive neurons,while in diabetic group we have found 40.74% ± 2.22% of n NOS immunoreactive perikarya(increase by 82.85 %). In turn in the pylorus we have observed 15.91% ± 0.58% n NOS containing neurons in control animals and 35.38% ± 1.54% in the diabetes group(increase by 122.37%). In the MG of the antrum and submucosal ganglion(SG) in the corpus hyperglycemia did not cause statistically significant changes. With regard to VIP-positive cell bodies in the antrum MG in the control animals we have noted 18.38 ± 1.39% and 40.74% ± 1.77% in the experimental group(increase by 121.65%). While in the corpus we have observed 23.20% ± 0.23% in the control and 30.93% ± 0.86% in the diabetes group(increase by 33.31%). In turn in the pylorus VIP positive cells bodies constituted 23.64% ± 1.56% in the control group and 31.20% ± 1.10% in the experimental group(increase by 31.97%). In the submucosal ganglion in the corpus we have noted 43.61% ± 1.06% in the control animals and 37.00% ± 1.77% in the experimental group(decrease by 15.15%). Expression of GAL-positive perikarya showed statistically significant changes only in the MG of the antrum and pylorus. In the antr

关键词： Immunohistochemistry Inhibitory neurons Streptozotocin Hyperglycemia Pig

Hepatoprotective effects of Nigella sativa seed extract against acetaminophen-induced oxidative stress

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Asian Pacific Journal of Tropical medicine 2016年第3期9卷 217-222页

作者： Gareeballa Osman Adam Md.Mahbubur Rahman Sei-Jin Lee Gi-Beum Kim Hyung-Sub Kang Jin-Shang Kim Shang-Jin Kim Department of veterinary medicine and Surgery College of Veterinary Medicine.Sudan University of Science and Technology Department of veterinary Pharmacology and Toxicology College of Veterinary Medicine.Chonbuk National University Korea Basic Science Institute Jeonju Center

Objective:To investigate the protective effects of Nigella sativa seed extract(NSSE) against acetaminophen(APAP)-induced hepaloloxicity in TIB-73 cells and ***:Toxicity in TIB-73 cells was induced with 10 μmol/L APAP and the protective effects of NSSE were evaluated at 25.50.75,100 μg/*** in rim examination,a total of 30 rals were equally divided into five experimental groups:normal control(vehicle),APAP(800 mg/kg body weight single IP injection) as a hepatotoxic control,and three APAP and NS pretreated(2 weeks) groups(APAP+NSSE 100 mg:APAP+NSSE 300 mg and APAP+NSSK 900 mg/kg).Results:TIB-73 cell viability was drastically decreased by(49.0±l.9)%after the 10 μmol/L APAP treatment,which also increased reactive oxygen species ***-treatment with NSSE at 25.50.75,and 100 μg/mL significantly improved cell viability and suppressed reactive oxygen species *** viro the APAP induced alterations in blood lactate levels,pH,anionic gap,and ion levels(HCO_3^-,Mg^(2+) and K^+),which tended to normalize with the NSSE *** NSSE also significantly decreased elevated serum levels of alanine aminotransferase,aspartate aminotransferase,lactate dehydrogenase,and alkaline phosphatase induced by APAP,which correlated with decreased levels of hepatic lipid peroxidation(nialondialdehyde),increased superoxide dismutase levels,and reduced glutathione *** hepatic histology was also found in the treatment groups other than APAP ***:The in vitro and in vim findings of this study demonstrated that the NSSE has protective effects against APAP-induced hepalotoxicity and metabolic disturbances by improving antioxidant activities and suppressing both lipid peroxidation and ROS generation.

关键词： Nigella sativa Acetaminophen Antioxidants Oxidative stress TIB-73 cells Rat

Baicalin inhibits colistin sulfate-induced apoptosis of PC12 cells

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Neural Regeneration Research 2013年第28期8卷 2597-2604页

作者： Hong Jiang Pengfei Lv Jichang Li Hongjun Wang Tiezhong Zhou Yingzi Liu Wei Lin Institute of veterinary medicine Northeast Agricultural University Institute of Animal Husbandry and veterinary medicine Liaoning Medical University Scientific Research Department Liaoning Medical Universit

Baicalin, a type of flavonoid extracted from the dried root of Scutellaria baicalensis georgi, has been shown to effectively inhibit cell apoptosis. Therefore, we assumed that baicalin would suppress colistin sulfate-induced neuronal apoptosis. PC12 cells exposed to colistin sulfate （62.5-500 μg/mL） for 24 hours resulted in PCl2 cell apoptosis. In addition, caspase-3 activity, lactate dehydrogenase level and free radical content increased in a dose-dependent manner. Subsequently, PC12 cells were pretreated with baicalin （25, 50 and 100 pg/mL）, and exposed to 125 pg/mL colistin sulfate. Cell morphology markedly changed, and cell viability increased. Moreover, caspase-3 activity, lac- tate dehydrogenase level and free radical content decreased. Results indicated that baicalin inhib- ited colistin sulfate-induced PC12 cell apoptosis by suppressing free radical injury, and reducing caspase-3 activity and lactate dehydrogenase activity.

关键词： neural regeneration traditional Chinese medicine baicalin colistin sulfate PC12 cells apoptosis caspase-3 lactate dehydrogenase grants-supported paper neuroregeneration

A survey of biochemical and acute phase proteins changes in sheep experimentally infected with Anaplasma ovis

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Asian Pacific Journal of Tropical Biomedicine 2018年第12期8卷 565-570页

作者： Zohreh Khaki Seyedeh Parastoo Yasini Seyedeh Missagh Jalali Department of Clinical Pathology Faculty of Veterinary MedicineUniversity of Tehran Department of Clinical Sciences Faculty of Veterinary MedicineIslamic Azad University Department of Clinical Sciences Faculty of Veterinary MedicineShahid Chamran University of Ahvaz

Objective: To determine the biochemical and acute phase proteins changes in sheep experimentally infected with Anaplasma ovis(A. ovis). Methods: One Iranian sheep naturally infected with A. ovis(parasitemia 0.02%) but with no other blood parasites based on blood smear and polymerase chain reaction methods was selected as donor, and it was splenectomized to induce high level of parasitemia. Then, three weeks after splenectomy when parasitemia was 6%, donor's blood was intravenously administered to each recipient animal. Five 5-6 months old Iranian male sheep without any blood parasites were selected as recipient animals. The percent of parasites, packed cell volume, serum biochemical parameters(urea, creatinine, bilirubin, aspartate aminotransferase activity, cholesterol, total protein, albumin, globulin, Fe), acute phase proteins(haptoglobin, total iron binding capacity, fibrinogen), were evaluated in sheep before and after being experimentally infected with A. ovis(until day 38). In addition, body weights of sheep were measured on days 0, 20 and 38. Results: In recipient sheep, microscopic examination of erythrocytes revealed a significant rise of parasitemia on days 12 and 15. The lowest level of packed cell volume in sheep was seen on day 15 post infection. A significant rise existed in mean urea and bilirubin(total, direct and indirect) on days 15 and 20. The increase of indirect bilirubin level was higher than direct bilirubin. Furthermore, serum Fe significantly increased on days 20 and 23. The mean total protein concentration significantly increased on day 38. A significant increase was found in the serum globulin concentration from days 20 and 27 to 38. Maximum values of haptoglobin were observed on days 27 and 30. Moreover, aspartate aminotransferase activity(from days 20-30) and cholesterol concentration(on day 20) significantly decreased. However, no significant changes were found in other parameters. Conclusions: Experimental ovine anaplasmosis caused by

关键词： Anaplasma ovis Acute phase protein Sheep Biochemical Iran