Tumor promoting effect of phenolphthalein on development of lung tumors induced by N-ethyl-N-nitrosourea in transgenic mice carrying human prototype c-Ha-ras gene

In order to examine tumor modifying effects of phenolphthalein (PhP), female transgenic mice carrying human prototype c-Ha-ras gene (rasH2 mice) were given a single intraperitoneal injection of 60 mg/kg body weight of N-ethyl-N-nitrosourea (ENU), followed by the diet containing 12,000 ppm PhP for 26-week. Histopathologically, alveolar hyperplasias, adenomas and adenocarcinomas were observed in the ENU + PhP group, but only hyperplasias and adenomas were observed in the ENU alone group. The incidence and multiplicity of adenocarcinomas in the ENU + PhP group was significantly increased as compared to that in the ENU alone group. The combined multiplicity of adenomas and adenocarcinomas in this group was also significantly higher than that of the ENU alone group. In addition, the ratio of area of adenomas in the ENU + PhP group was significantly higher than that in the ENU alone group. The result of our study suggests that PhP has a clear tumor promoting effect in the lung of rasH2 mice.     

Plasma Steroid Variations in Bull Calves Repeatedly Treated with Testosterone,Nortestosterone and Oestradiol Administered Alone or in Combination

The aim of this work was to investigate the secretion of dehydroepiandrosterone (DHEA), testosterone (T), dihydrotestosterone (DHT) and oestradiol (E) as biological markers in response to illegal administration of testosterone, 19-nortestosterone (N) and oestradiol, either alone or in combination. Twenty male Friesian calves (age 13-14 months) were allotted to a control group (n = 5), and five experimental groups (n = 3) each. Each experimental animal was repeatedly injected with one of the following hormonal treatments: E, T, N, T+E and N+E. Circulating DHEA, T, DHT and E were determined by radioimmunoassay. The administration of T alone did not induce any variation in plasma DHEA, T, DHT and E, which were similar to those in the control group. In contrast, DHEA, T and DHT were on average significantly lower in the T+E and N-treated groups (p < 0.01), whereas the administration of N+E resulted in the reduction of plasma T and DHT without any modification of plasma DHEA. The administration of E alone or in combination increased circulating levels of E but did not affect androgen plasma profiles. The results indicate that plasma levels of T do not permit detection of illegal treatments because plasma androgens always remained within the physiological range. Illegal E treatment could be detected in blood samples when they were collected at least every 20 days.     

AP液对微循环影响的试验研究

将实验小鼠分为A液(0.1％肾上腺素注射液)组、P液(2％硝酸毛果云香硷注射液)组、AP液(A液与P液等量混合液)组和生理盐水对照组,在麻醉状态下,于给药前和皮下注射给药后5、30、60min,应用显微录象、照相技术,定点观察和测量小鼠耳廓部细动脉、分枝毛细血管、毛细血管、集合毛细血管、细静脉等微血管的口径、血流速度、血管运动性和毛细血管开放数。结果,AP液能使各种微血管口径扩张,血流速度加快,血管运动性增强,毛细血管开放数增多,表明AP液有改善微循环的作用,而单用A液、P液,都使微血管收缩,血流速度快慢交替出现,血管运动性减弱,P液使毛细血管开放数增多,A液由减少到恢复。     

Adjuvant and immunostimulatory effects of a D-galactose-binding lectin from Synadenium carinatum latex (ScLL) in the mouse model of vaccination against neosporosis

Vaccination is an important control measure for neosporosis that is caused by a coccidian parasite, Neospora caninum, leading to abortion and reproductive disorders in cattle and serious economic impacts worldwide. A D-galactose-binding lectin from Synadenium carinatum latex (ScLL) was recently described by our group with potential immunostimulatory and adjuvant effects in the leishmaniasis model. In this study, we evaluated the adjuvant effect of ScLL in immunization of mice against neosporosis. First, we investigated in vitro cytokine production by dendritic cells stimulated with Neospora lysate antigen (NLA), ScLL or both. Each treatment induced TNF-α, IL-6, IL-10 and IL-12 production in a dose-dependent manner, with synergistic effect of NLA plus ScLL. Next, four groups of C57BL/6 mice were immunized with NLA + ScLL, NLA, ScLL or PBS. The kinetics of antibody response showed a predominance of IgG and IgG1 for NLA + ScLL group, whereas IgG2a response was similar between NLA + ScLL and NLA groups. Ex vivo cytokine production by mouse spleen cells showed the highest IFN-γ/IL-10 ratio in the presence of NLA stimulation for mice immunized with NLA + ScLL and the lowest for those immunized with ScLL alone. After parasite challenge, mice immunized with NLA + ScLL or ScLL alone presented higher survival rates (70-80%) and lower brain parasite burden as compared to PBS group, but with no significant changes in morbidity and inflammation scores. In conclusion, ScLL combined with NLA was able to change the cytokine profile induced by the antigen or lectin alone for a Th1-biased immune response, resulting in high protection of mice challenged with the parasite, but with low degree of inflammation. Both features may be important to prevent congenital neosporosis, since protection and low inflammatory response are necessary events to guide towards a successful pregnancy.     

连翘提取物对对乙酰氨基酚诱导小鼠肝损伤的保护作用

试验旨在探究连翘提取物（FSE）预防性干预对对乙酰氨基酚（acetaminophen,APAP）诱导小鼠肝损伤的保护作用及其潜在作用机制。采用半仿生-生物酶法提取连翘有效成分,使用APAP构建小鼠肝损伤模型。随机将60只雌性昆明小鼠分入正常对照（NC）组、APAP肝损伤模型（LD）组、连翘提取物（FSE）对照组、连翘提取物高剂量（HFSE＋LD）组、连翘提取物中剂量（MFSE＋LD）组和连翘提取物低剂量（LFSE＋LD）组,每组10只。HFSE＋LD组、MFSE＋LD组、LFSE＋LD组分别按每天200、100、50 μg/g灌胃给予连翘提取物,NC组和LD组分别灌胃等量生理盐水,每天2次,连续给药6 d。预防性给药3 d后,腹腔注射APAP,每天1次。末次给药12 h后,试验小鼠眼球采血并快速取出肝脏,检测血清中丙氨酸氨基转移酶（ALT）和天门冬氨酸氨基转移酶（AST）活性,以评价肝损伤程度;检测肝脏匀浆液中还原型谷胱甘肽（GSH）、超氧化物歧化酶（SOD）、丙二醛（MDA）和过氧化氢（H₂O₂）水平,以评价肝脏氧化应激程度;制作肝脏病理切片,经苏木精伊红（HE）染色后观察肝脏病理变化;采用探针药物法测定肝脏线粒体细胞色素P4502E1（CYP2E1）活性;采用实时荧光定量PCR检测肝脏线粒体CYP2E1 mRNA表达水平;采用Western blotting法检测肝脏CYP2E1蛋白表达情况。结果表明：与NC组相比,LD组小鼠血清中ALT、AST活性均显著增加（P<0.05）;肝脏SOD活性、GSH含量均显著降低（P<0.05）,MDA、H₂O₂含量,CYP2E1 mRNA及蛋白表达水平均显著升高（P<0.05）,成功构建小鼠肝损伤模型。200、100和50 μg/g的连翘提取物可显著降低肝损伤小鼠血清ALT和AST活性（P<0.05）,显著提高肝脏中SOD活性（P<0.05）;200、100 μg/g连翘提取物可显著提高肝脏中GSH水平（P<0.05）,显著降低肝脏中MDA、H₂O₂水平及CYP2E1的mRNA和蛋白表达量（P<0.05）。以上结果表明,连翘提取物可减轻APAP诱导的小鼠肝损伤程度且呈剂量依赖性,其潜在作用机制可能与所含活性物质的抗氧化作用以及对CYP2E1酶活性和表达的抑制有关。     

Enhancement of hepatocellular proliferative activity of kojic acid in mice by a simultaneous administration of ascorbic acid

To examine the tumor modification activity of kojic acid (KA) by sodium ascorbic acid (AA), 5-week-old male ICR mice were administered intraperitoneally with N-diethylnitrosamine (DEN) as an initiation treatment. Two weeks after the initiation treatment, animals were fed basal diet containing 0 (Group 1: DEN alone) or 3% KA (Group 3: DEN+KA), drinking water containing 5,000 ppm AA (Group 2: DEN+AA) or 3% KA and 5,000 ppm AA (Group 4: DEN+KA+AA) for 6 weeks. One week after the administration of KA and/or AA, all mice were subjected to two-thirds partial hepatectomy. At the end of the experimental period, all surviving mice were sacrificed and removed the liver. The liver weights of the Groups 3 and 4 were significantly increased, and the number of proliferating cell nuclear antigen positive hepatocytes and the gene expressions of Ccnc, Ccnd1, Ercc and Cyp7a1 were significantly increased in the Group 4, as compared to the Group 1. These results of the present study suggest that AA enhances the hepatocellular proliferative activity of KA in mice.     

结石通片对小鼠的急性毒性试验及其对乙二酰胺致大鼠肾结石的作用研究

[目的]研究结石通片对小鼠的急性毒性作用,对健康大鼠的利尿作用,以及对乙二酰胺致大鼠肾结石的调节作用.[方法]取ICR小鼠40只,雌、雄各半,随机分为对照组与给药组,每组20只,观察结石通片给药前后小鼠的体重变化和死亡情况;另取SD大鼠36只,雌、雄各半,分为对照组、低剂量组(给予5％结石通片溶液)和高剂量组(给予10...     

新疆野生荒漠肉苁蓉醇提物调节小鼠DCs成熟对Th1/Th2的免疫增强作用

旨在探讨新疆野生荒漠肉苁蓉醇提物(ethanol extracts of wild Cistanche deserticola,EEWCD)调节Th1/Th2免疫反应的特点及初步的作用机制.采用卵清白蛋白(ovalbumin,OVA)为抗原,研究EEWCD对小鼠体液免疫,细胞免疫,细胞因子分泌,树突状细胞(dendri...     

鸡血藤总黄酮对猪圆环病毒2型感染小鼠脾脏氧化应激的影响

试验旨在探讨鸡血藤总黄酮（TFSD）对猪圆环病毒2型（PCV2）感染小鼠脾脏氧化应激的影响。将70只昆明小鼠随机分为7组,对照组、TFSD组（100 mg/kg体重）、PCV2组、PCV2＋维生素C （VC）组、PCV2＋不同浓度TFSD （25、50和100 mg/kg体重）组,每组10只,连续3 d采用灌胃和腹腔注射PCV2病毒液的方法建立小鼠氧化应激模型,第4~6天每天按上述分别灌胃给予生理盐水、VC或TFSD。第7天剖杀小鼠,取脾脏分析黄嘌呤氧化酶（XOD）、髓过氧化物酶（MPO）和超氧化物歧化酶（SOD）活力,并检测还原型谷胱甘肽（GSH）和氧化型谷胱甘肽（GSSG）水平,计算GSH/GSSH。结果显示,PCV2感染小鼠后,脾脏中XOD与MPO的活力及GSSG水平显著上升（P＜0.05）,SOD活力、GSH水平及GSH/GSSG显著下降（P＜0.05）。TFSD处理小鼠脾脏的SOD活力、GSH水平和GSH/GSSG比值均显著高于PCV2组（P＜0.05）,而XOD、MPO活力和GSSG水平则显著低于PCV2组（P＜0.05）,对PCV2引起的氧化应激相关酶活力与相关分子水平变化的抑制作用优于VC。结果表明,鸡血藤总黄酮对PCV2诱导的小鼠脾脏氧化应激有良好的调节作用。     

Protective Effect of Sulforaphane on Reproductive Function of Male Mice with Cadmium Poisoning

The study was aimed to explore the protective effect of sulforaphane (SFN) on the reproductive function of male mice with cadmium poisoning.40 healthy clean grade male Kunming mice were randomly divided into four groups:control group (H₂O),cadmium chloride group (2.3 mg/kg CdCl₂),sulforaphane group (10 mg/kg SFN),sulforaphane + cadmium chloride group (10 mg/kg SFN+2.3 mg/kg CdCl₂),and continuous administration for 10 d,all mice were executed by dislocated cervical vertebra at 2 d after the last administration,and then the pathologic changes of testicular tissues,organ coefficient of testicle and epididymis,sperm quality and concentration of testosterone were tested.Additionally,the contents of GSH and MDA,and the activities of T-SOD in testis were also detected at the same time. Compared with the control group,pathology damages were observed in cadmium chloride group,organ coefficient of testis and epididymis,sperm quality and levels of testosterone extremely significantly decreased (P<0.01),the activities of T-SOD and GSH content were extremely significantly decreased (P<0.01),and the concentration of MDA was extremely significantly enhanced (P<0.01).Compared with the control group,the activity of T-SOD and concentration of GSH in sulforaphane group were significantly increased (P<0.05),and the concentration of MDA was not significant different between the control group and sulforaphane group (P>0.05).While compared with the cadmium chloride group,the sperm motility rate and sperm total count in sulforaphane and cadmium chloride group were extremely significantly increased (P<0.01),the organ coefficient of testicle and epididymis was increased significantly (P<0.05),the concentration of GSH and activity of T-SOD in testicular tissue were extremely significantly increased (P<0.01),and the concentration of MDA was extremely significantly decreased (P<0.01).The results indicated that sulforaphane had the protection effect on reproduction function of male mice with cadmium poisoning.     

莱菔硫烷对镉中毒雄性小鼠生殖机能保护作用的研究

本试验旨在研究莱菔硫烷(SFN)对染镉雄性小鼠生殖机能的保护作用.选择健康的清洁级雄性昆明系小鼠40只,随机分成4组,分别为对照组(H₂O)、镉组(2.3 mg/kg CdCl₂)、莱菔硫烷组(10 mg/kg SFN)、镉+莱菔硫烷组(10 mg/kg SFN+2.3 mg/kg CdCl₂),连续给药10 d,于最后一次给药2 d后脱颈处死,检测小鼠睾丸组织病理学变化、小鼠睾丸及附睾的脏器系数、精液品质及血清睾酮浓度.同时检测睾丸组织内丙二醛(MDA)、谷胱甘肽(GSH)含量及总超氧化物歧化酶(T-SOD)活性的变化.结果表明,与对照组相比,镉组小鼠的睾丸组织发生病理学损伤,睾丸及附睾的脏器系数、精液品质及血清睾酮浓度均极显著降低(P<0.01),睾丸组织T-SOD活性、GSH含量也极显著降低(P<0.01),MDA含量极显著提高(P<0.01);莱菔硫烷组T-SOD活性和GSH含量显著升高(P<0.05),MDA含量与对照组相比差异不显著(P>0.05).与镉组相比,镉+莱菔硫烷组小鼠的精子活率、精子总数极显著增加(P<0.01),睾丸与附睾的脏器系数显著增加(P<0.05),小鼠睾丸组织GSH含量、T-SOD活性极显著提高(P<0.01),MDA含量极显著降低(P<0.01).以上结果指出莱菔硫烷对镉中毒雄性小鼠的生殖机能具有保护作用.     

Effect of Total Flavonoids of Spatholobus suberectus Dunn on Oxidative Stress in Mice Spleen Induced by PCV2

The aim of this study was to investigate the effect of total flavonoids of Spatholobus suberectus Dunn (TFSD) on porcine circovirus type 2 (PCV2) induced oxidative stress in mice spleen.70 Kunming mice were divided into 7 groups:Control group, TFSD group (100 mg/(kg·BW)), PCV2 group, PCV2+vitamin C (VC) group, and PCV2+various concentrations of TFSD groups (25, 50 and 100 mg/(kg·BW)). Mice were continuously treated with PCV2 via both intragastric administration and intraperitoneal injection for 3 d to establish oxidative stress models. From the 4th to 6th day, mice were intragastric administrated with saline, VC or TFSD, respectively, according to the grouping method. At the 7th day, the activities of xanthine oxidase (XOD), myeloperoxidase (MPO) and superoxide dismutase (SOD), the levels of glutathione (GSH) and oxidized glutathione (GSSG), and the ratio of GSH to GSSG in the mice spleen were analyzed. The results showed that PCV2 infection significantly upregulated the XOD and MPO activities and GSSG content(P <0.05), and dramatically downregulated the SOD activity, GSH level and the ratio of GSH to GSSG (P <0.05) in the mice spleen.Compared to PCV2 group, the SOD activity, GSH content and the ratio of GSH to GSSG in mice treated with TFSD were significantly increased (P <0.05), while the activities of XOD and MPO and the level of GSSG were significantly decreased (P <0.05), showing better performance in the inhibition of PCV2 induced changes of oxidative stress associated enzyme activities and moledule levels than VC.In conclusion,TFSD had regulative effect on the oxidative stress induced by PCV2 in mouse spleen.     

橄榄苦苷对小鼠脑缺血再灌注损伤的保护作用

本研究为探讨橄榄苦苷联合依达拉奉对小鼠脑缺血再灌注损伤的影响及其保护作用机制,将50只健康小鼠分成假手术组、模型组、橄榄苦苷组、依达拉奉组、橄榄苦苷＋依达拉奉组。利用双侧颈总动脉结扎方法制备慢性脑缺血再灌注小鼠模型,造模后药物处理21 d,用放射免疫法检测脑组织肿瘤坏死因子α（TNF-α）、白细胞介素1（IL-1β）、白细胞介素10（IL-10）含量,用比色法检测脑组织ATP酶、髓过氧化物酶（MPO）、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活性及丙二醛（MDA）含量,用免疫组织化学法及Western blotting法检测大脑皮层脑源性神经营养因子（BDNF）的表达。结果显示,与假手术组相比,模型组脑组织TNF-α、IL-1β、MDA的含量及MPO活性极显著升高（P<0.01）,IL-10、ATP酶、SOD、CAT水平均极显著降低（P<0.01）,大脑皮层BDNF表达水平极显著降低（P<0.01）。与模型组比较,橄榄苦苷或依达拉奉治疗后,脑组织TNF-α、IL-1β、MDA的含量及MPO活性极显著降低（P<0.01）,IL-10、ATP酶、SOD、CAT水平均极显著升高（P<0.01）,大脑皮层BDNF表达水平极显著升高（P<0.01）。橄榄苦苷和依达拉奉联合治疗后脑缺血再灌注损伤的恢复更加显著。综上所述,橄榄苦苷和依达拉奉对小鼠脑缺血再灌注损伤具有明显的保护作用,其机制可能与改善神经功能、减少自由基损伤和抑制炎症因子水平有关,橄榄苦苷和依达拉奉联合治疗可以发挥更好的作用。     

兔IL-6基因真核表达载体的构建及其对pcDNA-VP60DNA疫苗佐剂效应研究

为研究兔白介素6（interleukin6,IL-6）真核表达质粒（pcDNA-IL-6）对核酸疫苗免疫效果的影响,本文构建了真核表达质粒pcDNA-IL-6,并将其与核酸疫苗pcDNA-VP60联合免疫家兔,以pcDNA-VP60和质粒载体pcDNA3．1（＋）作对照,用血凝抑制试验检测试验兔体内特异性抗体水平。结果表明：真核重组质粒pcDNA-IL-6对重组质粒pcDNA-VP60均具有免疫增强作用,从免疫后7d到70d,pcDNA-VP60／pcDNA-IL6联合免疫组抗体水平均高于pcDNA-VP60免疫组,差异具有显著统计学意义。     

Approaches to the identification of non-essential genes of African swine fever virus.

It is poorly understood why vaccines could not be developed for the control and prevention of African swine fever (ASF) virus infection. The aim of our study was to identify genes non-essential for ASF virus replication because there were indications that certain viral gene products, which apparently are non-essential for viral replication, conferred protection from death due to ASF. A cosmid library representing the genome of ASF virus strain France 64 was established and characterized. Then, in order to inactivate viral genes by insertion, the beta-galactosidase (beta-gal) gene was introduced either randomly or at specific locations of selected cloned DNA fragments. These constructions were transfected into cells which had been previously infected with a cell-culture-adapted viral strain in order to allow the generation of recombinant progeny virus. Viable recombinant progeny was identified by at least one of the following means: (1) expression of beta-gal; (2) detection of beta-gal specific DNA by plaque hybridization, and (3) absence of a functional product of the inactivated gene. Presently, we are characterizing a recombinant virus with an insertionally inactivated thymidine kinase gene.     

Protective Effects of Oleuropein on Cerebral Ischemia-reperfusion Injury in Mice

To explore the protection role and mechanism of oleuropein and edaravone on cerebral ischemia-reperfusion injury in mice.Fifty mice were divided into five groups:Sham operation group,model group,oleuropein group,edaravone group,oleuropein+edaravone group.The model was established by ligating common carotid artery.After modeling,the mice were administrated with oleuropein,edaravone and oleuropein+edaravone for 21 d,respectively.The contents of TNF-α,IL-1β and IL-10 were detected by radioimmunoassay.The activities of ATPase,MPO,SOD and CAT and MDA content were measured by spectrophotometry.The changes of BDNF expression in cerebral cortex were analyzed by immunohistochemistry and Western blotting.The results showed that compared to sham operation group,the contents of TNF-α,IL-1β,MDA and MPO activity in model group were extremely significantly increased (P<0.01),and the levels of IL-10,ATPase,SOD and CAT in model group were extremely significantly reduced (P<0.01),the BDNF expression in model group was extremely significantly decreased (P<0.01).Compared to model group,the contents of TNF-α,IL-1β,MDA and MPO activity in oleuropein,edaravone,and oleuropein+edaravone groups were extremely significantly decreased (P<0.01),the levels of IL-10,ATPase,SOD and CAT were extremely significantly increased (P<0.01),the BDNF expression was extremely significantly up-regulated in cerebral cortex of treatment group (P<0.01).Furthermore,the oleuropein+edaravone combined administration showed a better effect.The treatments of oleuropein and edaravone had a protective effect on cerebral ischemia reperfusion injury,the mechanisms of which might depend on improving neurological function,reducing free radical lesion and inhibiting inflammatory response.Moreover,the oleuropein+edaravone combination therapy might have an additive effect.     

芪参口服液的毒理学研究

为评价芪参口服液临床用药的安全性,根据毒理学评价程序和方法对其进行了急性毒性试验和长期毒性试验。在急性毒性试验中,将40只昆明小鼠按体重随机分为2组,采用最大给药量法测定小鼠口服芪参口服液的最大耐受剂量;在长期毒性试验中,将80只Wistar大鼠按体重随机分为4组,3个剂量组分别按3.75、7.5、15 g/kg体重给大鼠灌胃,对照组给予同体积生理盐水,1次/d,连续4周。记录每日饮水量、饲料采食量及每周体重,检测给药4周及停药2周血液生化指标、血常规、脏器系数和组织病理学变化。试验结果显示,小鼠口服芪参口服液的最大耐受剂量为生药量40 g/kg,此剂量相当于临床日用量的40倍。部分给药组采食量和饮水量与对照组相比出现显著(P＜0.05)或极显著(P＜0.01)变化,但对平均周增重无显著性影响(P＞0.05);血常规的部分指标出现显著(P＜0.05)或极显著变化(P＜0.01),但停药2周后,所有指标与相应的对照组相比无显著性差异(P＞0.05);给药组血液生化和脏器系数的各项指标与对照组相比,无显著性变化(P＞0.05)。病理组织学检查无明显差异。试验结果表明,在本次试验条件下,该口服液安全无毒。     

羊口疮病毒B2L、F1L基因融合真核表达质粒诱导小鼠的免疫应答及IL-2对其作用影响的研究

为评价羊口疮病毒(OrfV)B2L、F1L基因融合真核表达质粒pVAX1-B2L-F1L免疫小鼠诱导的体液和细胞免疫应答及IL-2对其免疫作用的影响。本研究将构建的pVAX1-B2L-F1L真核质粒转染MDBK细胞后,采用RT-PCR和间接免疫荧光试验(IFA)检测B2L-F1L融合基因在MDBK细胞中的表达;将pVAX1-B2L-F1L、pVAX1-B2L-F1L+pVAX1-IL-2、pVAX1空载体、生理盐水对照组KM系小鼠通过后腿肌肉注射的方式免疫,采用ELISA方法检测免疫小鼠血清中OrfV特异性抗体以及Th1型(IL-2、IFN-γ)、Th2型(IL-4、IL-6)细胞因子;MTT法检测小鼠脾淋巴细胞增殖反应。结果显示,pVAX1-B2L-F1L重组质粒能够在MDBK细胞中表达;pVAX1-B2L-F1L+pVAX1-IL-2联合免疫组小鼠血清抗体及IL-2和IFN-γ水平均显著高于pVAX1-B2L-F1L组;该联合免疫组小鼠血清IL-4、IL-6细胞因子水平与pVAX1-B2L-F1L组相比差异不显著(p>0.05);该联合免疫组小鼠的脾淋巴细胞增殖水平高于pVAX1-B2L-F1L组(p<0.05)。由此表明,pVAX1-B2L-F1L能够诱导小鼠产生OrfV特异性体液免疫和细胞免疫应答,联合pVAX1-IL-2诱导的免疫反应以细胞免疫应答为主,且能够促进Th1型细胞因子的分泌。本研究为OrfV基因工程疫苗的研制提供了参考依据。     

猪圆环病毒2型诱导小鼠体内免疫细胞氧化胁迫模型的建立

试验旨在建立猪圆环病毒2型（PCV2）诱导的小鼠体内免疫细胞氧化胁迫模型。筛选PCV2感染剂量及感染时间,采用腹腔注射、滴鼻和灌胃3种途径联合方式于第1、2、3 天或第1、3、5、7天给予昆明系小鼠感染PCV2病毒原液或10^－1 PCV2病毒液,分别于感染后7、14、21 d剖杀小鼠,测定活性氧（ROS）、总谷胱甘肽（T-GSH）、还原型谷胱甘肽（GSH）、氧化型谷胱甘肽（GSSG）水平及黄嘌呤氧化酶（XOD）、髓过氧化物酶（MPO）、诱导型一氧化氮合酶（iNOS）活性,探讨PCV2感染时间与活性氧水平变化的关系,建立免疫细胞氧化胁迫动物模型。结果显示,第1、2、3天每天经3种途径联合感染PCV2病毒原液,1 mL/只,为后续试验感染最佳方案。PCV2感染小鼠3个时间点细胞内ROS水平较空白对照组均极显著升高（P < 0.01）,感染后7、14 d GSH水平显著降低（P < 0.05）;感染后7 d GSSG水平极显著高于空白对照组（P < 0.01）;感染后7 d T-GSH水平显著低于空白对照组（P < 0.05）,感染后14 d T-GSH水平极显著低于空白对照组（P < 0.01）。感染后7 d脾脏XOD、MPO、iNOS活性与空白对照组相比均存在显著差异（P < 0.05）。结果表明,PCV2成功感染小鼠,试验感染方案为第1、2、3 天每天经3种途径联合感染PCV2病毒原液,1 mL/只,且用PCV2病毒原液感染7 d是建立小鼠体内免疫细胞氧化胁迫模型的最佳条件。     

Establishment of Model for Oxidative Stress in Mice Immune Cells Induced by PCV2 in vivo

The aim of this experiment was to establish the oxidative stress model of immune cells in mice induced by PCV2. Optimal infection titer and time of PCV2 were selected. On the 1st,2nd and 3rd day or 1st, 3rd,5th and 7th day,Kunming mice were treated with 10⁰ PCV2 or 10^-1 PCV2 by 3 ways (intraperitoneal injection, intranasal administration and intragastrical administration). 7, 14, 21 d post administration, we killed the mice. Reactive oxygen species (ROS),total glutathione (T-GSH),reduced glutathione (GSH),oxidized glutathione (GSSG) levels and xanthine oxidase (XOD), myeloperoxidase (MPO) and inducible nitric oxide synthase (iNOS) activity were determined to investigate the relationship between the infection time point and the change of ROS level after PCV2 infection. Thus to establish mice immune cells oxidative stress model. The results showed that infection with 10⁰ PCV2 virus in 3 ways every day both at day 1, 2, 3 and 1 mL per mouse was the best program for the follow-up experiment. The intracellular level of ROS of mice infected with PCV2 was extremely significantly increased both at 7, 14 and 21 d (P < 0.01);7 and 14 d post infection, GSH level of PCV2 infected mice had significant difference compared with that of control group (P < 0.05). 7 d after infection, GSSG level was extremely significantly higher than that of control group (P < 0.01); 7 d after infection, T-GSH level was significantly lower than that of control group (P < 0.05), T-GSH was extremely significantly lower than that of control group at 14 d post infection (P < 0.01). XOD, MPO and iNOS activity between PCV2 infection group and blank control group were significantly different at 7 d post infection (P < 0.05). It suggested that PCV2 successfully infected mice, experimental infection programme was Kunming mice were treated with 10⁰ PCV2 by three ways (intraperitoneal injection, intranasal administration and intragastrical administration) both at the 1st,2nd and 3rd day of the experiment.The best condition to establish the oxidative stress model of immune cells in mice was using 10⁰ PCV2 to infect for 7 days.