中药方剂中复合多糖的免疫活性观察

本试验所研究的中药方剂是疗效确切、具有新疆地方特色的中兽药复方制剂, 对提高动物免疫力、降低畜禽的发病率和死亡率有确切、稳定的作用.     

唐古特莨菪中生物碱的分离、鉴定与生物活性研究

研究唐古特莨菪中生物碱对农业害虫(螨)的生物活性,可为利用该植物研制新型植物源杀虫(螨)剂提供理论依据。本文采用柱色谱分离法从唐古特莨菪地上部分分离得到2个生物碱类化合物,通过MS、NMR确定了其化学结构,分别为莨菪碱、东莨菪碱。采用微量点滴法和玻片浸渍法测定了莨菪碱、东莨菪碱对甘蓝蚜、桃蚜等9种蚜虫和朱砂叶螨、二斑叶螨等4种叶螨的触杀活性。结果表明,莨菪碱对禾谷缢管蚜、桃粉蚜、豆蚜和绣线菊蚜具有较强的触杀作用,LC₅₀值分别为257.863,275.459,344.645和344.717 mg/L。东莨菪碱对豆蚜和禾谷缢管蚜具有较强的触杀作用,LC₅₀值分别为311.585和392.309 mg/L。对照药剂鱼藤酮对甘蓝蚜和棉蚜具有较强的触杀作用,LC₅₀值分别为399.542和436.124 mg/L。对甘蓝蚜等9种蚜虫的触杀作用,莨菪碱强于或与东莨菪碱相当;对豆蚜、禾谷缢管蚜的触杀作用,莨菪碱和东莨菪碱均强于鱼藤酮;对甘蓝蚜、桃蚜和棉蚜的触杀作用,莨菪碱和东莨菪碱均弱于鱼藤酮。莨菪碱和东莨菪碱对截形叶螨、山楂叶螨、二斑叶螨和朱砂叶螨具有较强的触杀作用。2种生物碱对4种叶螨的毒力相当,对二斑叶螨和朱砂叶螨的毒力(201.027~224.172 mg/L)均高于截形叶螨和山楂叶螨(257.014~332.698 mg/L)。对照药剂鱼藤酮对截形叶螨、山楂叶螨的LC₅₀值分别为196.847和224.592 mg/L,其毒力高于莨菪碱和东莨菪碱。     

柴胡皂苷药理作用研究进展

综述了近年来柴胡皂苷在镇静、抗惊厥、解热、抗病毒、免疫调节、抗炎、保肝护肾以及抗肿瘤等多方面药理活性的研究近况,为近一步开发柴胡提供参考.     

A selected water-in-oil emulsion: Composition and usefulness as an immunological adjuvant

A selected water-in-oil (W/O) emulsion was evaluated in terms of stability and antibody-response and side-effects in experimental animals.The results from stability tests performed at 4°C and 37°C indicate a storability of the W/O emulsion of more than a year under normal storage conditions. The antibody-response of rabbits after injection of E.coli K99 antigen and bovine serum albumin (BSA) using the W/O emulsion as vehicle was generally significantly higher than the response to the same antigens injected within incomplete Freund's adjuvant (IFA): twenty days after the first antigen injection the rabbits injected with antigen in the W/O emulsion showed average anti-K99 and anti-BSA titres which were ca 5x as high as the titres in sera from rabbits injected with the antigens in IFA. Thirty-two and forty-two days after the first antigen injections the W/O group showed anti-K99 titres which were respectively ca 3.5x and 2x higher than titres in the IFA group, while anti-BSA titres in the W/O group were respectively ca 13x and 6.5x higher than in the IFA group.Intra-muscular and/or subcutaneous injection of the W/O emulsion in rabbits and pigs resulted in a slight degeneration of local tissue. When antigens were present in the emulsion granulomatas were often observed. The rise in temperature in pigs after injection of killed E.coli 0149:K91,K88 in the W/O emulsion was comparable with that induced by E.coli in physiological saline.From these results it is concluded that the W/O emulsion is likely to be a promising basis for veterinary vaccines and that the application of the W/O emulsion may contribute to the limitation of the number of animals used for raising antibodies for general immunological purposes.     

三七皂苷药理作用研究进展

综述了近年来三七皂苷在血液系统、心脑血管系统、神经系统、免疫系统、物质代谢系统,以及抗炎、抗衰老、抗肿瘤等多方面生理活性的研究近况,为进一步开发三七提供参考.     

苦荞麦中芦丁提取及体外抑菌活性研究

采用乙醇回流法提取苦荞麦中的芦丁,并进行TLC检测、紫外光谱和红外光谱检测。将芦丁对两种供试菌进行体外最小抑制浓度(MIC)、最低杀菌浓度(MBC)、半数抑菌浓度(IC50)测定。结果表明,其对大肠埃希氏杆菌、金黄色葡萄球菌的MIC分别为0.40mg/mL和0.20mg/mL;MBC分别为0.50mg/mL和0.20mg/mL;IC50分别为0.2605mg/mL和0.1607mg/mL。     

田七总皂苷的制备及其免疫活性试验

用700mL/L乙醇回流提取、大孔吸附树脂分离纯化得到田七总皂苷(PNS),进行薄层色谱鉴别和含量测定。为评价PNS的免疫调节作用,用环磷酰胺(CTX)为免疫抑制剂建立小鼠免疫抑制模型,分别以4种剂量的PNS(5、25、50、75mg/kg)腹腔注射给药,测定胸腺和脾脏指数以及血清中IL-2、IFN-γ和TNF-α水平,并从组织形态上观察胸腺与脾脏的结构变化。结果表明,PNS能使小鼠胸腺和脾脏指数有所提高,但无统计学意义(P>0.05);25mg/kg剂量能显著升高血清中IFN-γ和TNF-α水平(P<0.05),50mg/kg剂量能显著升高血清中TNF-α水平(P<0.05);PNS处理组能不同程度地对抗模型所致小鼠胸腺和脾脏的萎缩,增加胸腺皮质厚度和细胞数量,增加脾小结大小和淋巴细胞数量,在高剂量组尤为明显。     

Studies about the adjuvant activity of saponin fractions in foot and mouth disease vaccine. III. Comparison of the irritant, adjuvant and hemolytic activities of six commercial saponins and their hemolytic fractions obtained by chromatography on sephadex G 100.

12 experimental vaccines were prepared to compare the irritant and adjuvant activity in cattle of 6 commercial saponin preparations and their hemolytic fractions. It is still not known if a single substance is responsible for the irritant, adjuvant and hemolytic activities of the saponin preparations. The quantities of saponin added were standardised on the base of a constant hemolytic activity rather than on a weight of powder per dose of vaccine base. A FMD vaccine was used to reveal the adjuvant activity. It was concluded that the irritation is related to the hemolytic activity and not to the weight of powder. Irritation is slightly reduced when a toxic effect appears. The adjuvant activity was higher for untreated saponin preparations with high hemolytic activity used at low dose and for one of the chromatographic saponin fractions. The adjuvant activity is reduced when toxic effect appear. Toxicity of less hemolytic saponins used at high dose is removed by chromatography. Highly hemolytic saponins used at low dose become toxic after chromatographic treatment.     

猪阴道乳酸菌分离及益生功能评价

旨在从健康母猪阴道内分离筛选益生乳酸菌,为防治猪产后子宫内膜炎提供益生菌株。测定乳酸菌的抗菌活性、产酸、过氧化氢能力及抗生素敏感性。研究结果表明,8株菌株可抑制3种病原菌生长,并具有良好的产酸性能,LTL1和LTL2不能够产生过氧化氢,所有菌株均对庆大霉素具有抗性。LTL1和LTL10抑菌物质为有机酸类和蛋白类。8株乳酸菌可作为潜在的益生菌用于防治母猪子宫内膜炎。     

脑心肌炎病毒灭活疫苗佐剂的筛选及其效力评价

采用ISA201、ISA15A、GEL、CP940和1313佐剂分别与脑心肌炎病毒(EMCV)灭活抗原混合,配制成5种疫苗,采用BALB/c小鼠进行免疫保护试验。结果显示,5种佐剂疫苗对小鼠无明显不良反应,均能够诱导机体产生抗EMCV特异性ELISA抗体、中和抗体和淋巴细胞增殖反应,其中ISA15A、ISA201和GEL佐剂组抗体水平和淋巴细胞增殖转化刺激指数都显著高于抗原对照组(P0.01),而且,ISA15A佐剂组免疫应答反应高于ISA201和GEL佐剂组(P0.05)。采用EMCV强毒攻击,ISA15A、ISA201和GEL佐剂组免疫保护率均达90%,表明用ISA15A、ISA201和GEL这3种佐剂配制的EMCV灭活疫苗具有较高的免疫疫保护效力,为该病灭活疫苗研制奠定基础。     

Isolation of Streptococcus agalactiae and an aquatic birnavirus from doctor fish Garra rufa L

Background

The doctor fish, Garra rufa, has become increasingly popular as a treatment for skin disorders and for pedicures in recent years. Despite this there is very little information available regarding the welfare of these fish and the range of potential pathogens they may carry. In this study, a group of fish suffering from post-transport mortalities were examined and the isolated pathogens identified.

Findings

Group B Streptococcus agalactiae was isolated from kidney swabs of the fish and found to be resistant to a number of antibiotics. In addition to this, a fish virus belonging to the aquabirnavirus group, serogroup C was isolated for the first time in Ireland. However, no clinical signs of disease typical of bacterial or viral infections were observed in any fish examined.

Conclusions

As no clinical signs of disease attributable to either of the pathogens identified were found it was concluded that the mortalities were most likely due to transport related stress exacerbated by the presence of the pathogens. Further work is required to assess the suitability of current transport strategies and to examine the potential risk associated with the transport of live ornamental fish.     

Keratoconjunctivitis sicca: immunological evaluation of 62 canine cases

The etiology of keratoconjunctivitis sicca (KCS) in 62 dogs was evaluated, using immunologic techniques. Using direct fluorescent antibody testing, autoantibodies within the lacrimal, salivary, or pancreatic glands were detected in 5 of 8 dogs tested. Circulating antibodies to the nictitating membrane gland, main lacrimal gland, parotid salivary gland, or mandibular salivary gland were detected using indirect fluorescent antibodies in 9 of 31, 3 of 31, 5 of 31, and 5 of 31 sera, respectively. Using radial immunodiffusion, hyper-gamma-globulinemia was detected in 21 of 30 dogs with KCS. Antinuclear antibodies, primarily in a nucleolar pattern, were demonstrated in 20 of 50 dogs with KCS. Lymphocytic infiltrates were evident in 5 of 9 labial salivary biopsies, 2 of 4 parotid gland specimens, 2 of 4 mandibular gland specimens, and 2 of 3 thyroid gland specimens taken from dogs with KCS. Autoimmune diseases had been previously documented in 4 of 62 dogs. Twenty-five of the 62 dogs (40%) had concurrent problems indicative of an underlying immunologic disorder.     

Development of a water-in-oil-in-water adjuvant for foot-and-mouth disease vaccine based on ginseng stem-leaf saponins as an immune booster

There has been an increasing interest in finding new formulations that qualify as vaccine adjuvants, which must be safe, stable, and have the capacity to stimulate a strong immune response. In this study, a basic formulation of a water-in-oil-in-water (W/O/W) adjuvant CV13 was developed, and ginseng stem-leaf saponins (GSLS) were added as an immune booster into oil phase. The physicochemical properties of the adjuvant were tested. Furthermore, the immune activity and the adjuvant effects, as indicated by the foot-and-mouth disease virus (FMDV) antigen were evaluated. The results showed that CV13 was similar in appearance to ISA 206 and could package FMDV antigen into a stable W/O/W emulsion. The FMD vaccine prepared with CV13 alone or CV13 containing GSLS achieved pharmaceutical characteristics comparable to a vaccine prepared with ISA 206, moreover the structural stability of the CV 13 vaccine was found to be better. Mice that were immunized with the FMD vaccine prepared with CV13 containing GSLS presented a significantly higher LPBE antibody titer and splenocyte proliferation rate than those immunized with a vaccine prepared with CV13 alone (p < 0.05). In addition, there was no significant difference between the groups that were immunized with FMD vaccine prepared with CV13 containing GSLS and ISA206 in terms of cellular and humoral immune response. In this paper, CV13 containing GSLS shows excellent immunologic adjuvant effect in mice model, and this new adjuvant may provide a potential choice for FMD vaccine production in the future.     

大片吸虫组织蛋白酶L全长序列的获取及其生物信息学分析

根据文库载体序列与原组织蛋白酶表达序列标签（EST）设计引物,以大片吸虫cDNA文库为模板,进行Touchdown PCR与梯度PCR相结合的扩增反应,扩增产物克隆人pMD18-T载体。经鉴定后测序,并采用生物信息学技术对序列进行开放阅读框（ORF）、编码氨基酸序列、蛋白质同源性比较、二级、三级结构等分析;结果所获序列全长990bp,编码330个氨基酸,分子量36771.2u,等电点5．44;具有较明显的螺旋、片层和无规卷曲等二级结构,α螺旋占17．3％,β折叠占27．0％,无规卷曲占55．8％;具有2个较明显的跨膜区和2个疏水区,未发现明显的信号肽和糖基化位点;氨基酸序列同源性比对显示其属于半胱氨酸家族;三级结构同源建模预测显示其与组织蛋白酶K（PDB number 2f7dA）有49％的一致性。     

江苏省羊口疮病毒的分离鉴定及B2L基因的遗传进化分析

为了解江苏省近年来羊口疮病毒（Orf virus,ORFV）的流行情况,更好地控制江苏地区的羊口疮病,2013~2015年采集江苏部分地区羊口疮疑似病料121份,进行病毒分离鉴定及其B2L基因的遗传进化分析。结果显示,样品中ORFV PCR检测阳性4份,用胎羊鼻甲骨细胞（OFTu）和MDBK细胞进行病毒分离,分离到4株病毒。这4株病毒分别命名为ORFV/Ovis/XZ/Jiangsu/2015/China、ORFV1/Ovis/DT/Jiangsu/2015/China、ORFV2/Ovis/DT/Jiangsu/2015/China和ORFV/Ovis/SL/Jiangsu/2015/China。扩增ORFV的B2L基因全长并绘制遗传进化树。B2L基因序列分析显示,4株分离株之间的核酸同源性为98.5%~100.0%。遗传进化树显示,ORFV/Ovis/XZ/Jiangsu/2015/China株、ORFV1/Ovis/DT/Jiangsu/2015/China株和ORFV2/Ovis/DT/Jiangsu/2015/China株与SC-JY、GX-YB、JS-FX株聚成一簇,核苷酸同源性为97.8%~100.0%。ORFV/Ovis/SL/Jiangsu/2015/China与LiaoNing、HuB、Gansu株亲缘关系接近,核苷酸同源性为98.8%~98.9%。4株分离株与中国疫苗株的核苷酸同源性为96.8%~98.1%。结果表明,江苏地区的ORFV来源可能不同,有必要开展更为深入的流行病学调查,为防控江苏地区的羊口疮奠定基础。