低能量氦氖激光照射对荷瘤小鼠NK细胞活性的影响

研究低能量氦氖激光照射荷瘤小鼠对ＮＫ细胞活性的影响。选取ＢＡＬＢ／Ｃ雄性小鼠腹腔接种Ｓ１８０,接种后进行激光照射小鼠内眼角,监测了正确对照组,肿瘤对照组和７．３３、１１．００、１４．６７、２２．００、２９．３３Ｊ／ｃｍ＾２五个激光照射线能明显提高荷瘤小鼠ＮＫ细胞活性;７．３３、２９．３３Ｊ／ｃｍ＾２照射组则长高不明显。     

氦氖激光合用环磷酰胺对S180荷瘤鼠脾脏淋巴细胞IL-2诱生活性和增殖反应的影响

以S180腹水瘤小鼠为动物模型，应用11.00、14.67和22.00J/cm^2 3种剂量氦氖激光照射荷瘤鼠哈德腺，同时联合应用化疗药物环磷酰胺，对S180荷瘤鼠脾脏淋巴细胞IL-2诱生活性和淋巴细胞增殖反应的影响作了系统地研究。结果表明：CYT/氦氖激光照射联合应用组IL-2诱生活性和淋巴细胞增殖反应均明显高于CYT对照组。CYT可对荷瘤体产生某种程度的免疫抑制效应，而氦氖激光照射则可在某种程度上改善这一效应。     

"神蜂"牌J9311蜂胶囊抗癌作用试验研究

以J931 1蜂胶囊水溶液为样品 ,对小鼠进行了抑制S1 80 实体瘤 (小鼠肉瘤细胞系 )和H2 2 实体瘤 (小鼠肝癌细胞系 )试验以及成分合理性分析试验。试验结果显示 ,对于S1 80 实体瘤 ,J931 1蜂胶囊的有效抑瘤剂量范围是 :0 6g kg～ 4 8g kg。以0 3g kg、0 6g kg、1 2g kg为低、中、高剂量进行抑瘤试验显示 ,中、高剂量组小鼠瘤重明显降低 ,与对照组比较差异显著 (P <0 0 5 ,P <0 0 1 ) ;对于H2 2 实体瘤的有效抑瘤剂量范围是 :1 6g kg～ 6 0g kg ,重复试验差异性结果与S1 80 实体瘤相同。结果表明J931 1蜂胶囊具有抑制小鼠S1 80 和H2 2 实体瘤的作用。J931 1蜂胶囊成分合理性试验显示 ,J931 1蜂胶囊组分抑瘤效果显著优于单独成分及各成分二二组合 ,单独成分中蜂王浆和蜂胶起主要抑瘤作用 ,中药成分起辅助作用     

重离子12C6+治疗小鼠移植性肿瘤S180的试验研究

从实验肿瘤学、肿瘤病理学等角度对碳离子治疗小鼠移植性肿瘤S180进行了研究，结果表明，在有氧和急性缺氧情况下碳离子可有效治疗小鼠移植性肿瘤，碳粒子照射后主要引起肿瘤组织的坏死变化。     

Hepatitogenicity of three plaque purified mutants of hepatotropic mouse hepatitis virus, MHV-2.

Hepatitogenicity of three plaque purified mutant strains of mouse hepatitis virus, designated as MHV-2S, -2M and -2L, isolated from MHV-2 infected SR-CDF1-DBT cells was studied. After intraperitoneal inoculation with 2 x 10(5) PFU of parental MHV-2 and its mutants to 4-week-old female ICR mice, 40% of mice inoculated with MHV-2S and 20% of mice with -2M died in one week, whereas with -2L all mice survived. All mice inoculated with MHV-2 died in 3 days postinoculation (p.i.). Virus titer of the liver of mice inoculated with MHV-2, -2S and -2M reached peaks (MHV-2:10(7) PFU/0.2 g, -2S: 10(5) PFU/0.2 g and -2M: 10(6) PFU/0.2 g) at 96 hr p.i., while with -2L a peak titer (10(3) PFU/0.2 g) was shown at 48 hr p.i. Immunofluorescence revealed MHV specific antigen in the liver of MHV-2S infected mice in and around necrotic areas though less extensive than that of parental MHV-2 infected mice. With MHV-2M specific fluorescence was restricted in degenerated hepatocytes in the small necrotic foci. In mice inoculated with MHV-2L only faint fluorescence was detected. Histopathologically, in the liver of MHV-2S infected mice zonal necrosis and cell infiltration were observed. There were spotty necrosis and focal cell infiltration in the liver of MHV-2M infected mice and only small inflammatory foci were seen in MHV-2L infected mice. Large number of extracellular virions were detectable in MHV-2S but not in -2M and -2L infected mice by electron microscopy.     

Co-delivery of IL-2 or liposomes augment the responses of mice to a DNA vaccine for pseudorabies virus IE180

Recently we have demonstrated, with a DNA vaccine, that the immediate early protein (IE180) of pseudorabies virus provides a moderate level of protection in mice. In order to improve its immunogenicity and protective capacity, this IE180 DNA vaccine was delivered to C3H/HeJ mice either in combination with an IL-2 expressing plasmid or complexed with cationic liposomes. Co-delivery of the vaccine and IL-2 DNA by gene gun resulted in seroconversion in 5/5 of the vaccinated mice after a single administration, whereas two intramuscular (i.m.) injections were required to achieve seroconversion in all mice. Antibody and delayed-type hypersensitivity responses were augmented in mice, which received the DNA vaccine and the IL-2 gene compared to those of mice receiving the DNA vaccine alone. In addition, the time of death after challenge was significantly delayed in mice, which received the IL-2 gene. The proportion of surviving mice (40%), however, was similar to that obtained in mice which received the vaccine alone by gene gun. Liposome-mediated vaccine delivery also resulted in a higher rate of seroconversion when compared with that induced by the naked DNA vaccine. Thus, all vaccinated mice seroconverted after either two i.v. or three i.m. injections of the liposome/DNA complex, with 40 and 25% of these mice being protected against challenge, respectively. These data support that co-administration of the IE180 DNA vaccine with the IL-2 gene or delivery in liposomes are two effective approaches to increase its immunogenicity.     

猪霍乱沙门菌C500运载CSFV新型基因疫苗的稳定性及免疫安全性

为研究猪霍乱沙门菌C500(S.C500)运载猪瘟病毒(CSFV)新型基因疫苗的稳定性与免疫安全性,将表达载体pCB-ME2-IL-15、pCC-ME2-IL-15、pCI-ME2-IL-15及pCI-neo转入S.C500,采用体外增殖试验考察不同表达载体在S.C500中的稳定性;采用携质粒S.C500对ST细胞的粘附、侵袭力和增殖试验研究不同表达载体对运载体生物学特性的影响;采用BALB/c小鼠口服免疫试验研究表达质粒在体内的稳定性和运载体对小鼠的安全性。结果显示:体外增殖中pCB-ME2-IL-15、pCC-ME2-IL-15和pCI-ME2-IL-15稳定性好于pCI-neo,说明外源目的基因正调表达质粒在运载体内的稳定性;空运载体S.C500对ST细胞的粘附率和侵袭率均高于4组含不同质粒的菌株,而4组含不同质粒的菌株之间亦有差异;空运载菌在ST细胞内的增殖能力与4组含不同质粒的菌株之间亦有显著性差异;分别对携带表达载体pCB-ME2-IL-15、pCC-ME2-IL-15和pCI-ME2-IL-15菌株以每只2×108CFU/0.2 mL的剂量进行小鼠口服免疫,腹泻率依次为0%、8.33%和25%。在第7和14天,使用DHL/Amp琼脂培养平板,均可从肝脏、脾脏、十二指肠和新鲜粪便样品中分离到重组菌。试验提示采用猪霍乱沙门菌C500运载CSFV新型基因疫苗免疫BALB/c小鼠具有相对稳定性和免疫安全性。     

Oral vaccination with attenuated Salmonella enterica serovar Typhimurium expressing Cap protein of PCV2 and its immunogenicity in mouse and swine models

Attenuated Salmonella enterica serovar Typhimurium (S. typhimurium) was selected as a transgenic vehicle for the development of oral vaccines against Porcine circovirus type 2 (PCV2). The Cap-encoding gene of PCV2 was amplified by PCR and cloned into expression vector pYA3341. The recombinant plasmid pYA3341-Cap was transformed into attenuated S. typhimurium X4550. BALB/c mice were inoculated orally with various doses of attenuated S. typhimurium X4550/pYA3341-Cap. The bacterium was safe to mice at dose of 2×10(9)cfu and eventually eliminated in the spleen and mesenteric lymph nodes at 4 weeks post-immunization. The flow cytometry analysis showed that the percentage of CD4(+) T cells and CD4(+)/CD8(+) ratio were increased significantly in mice immunized with attenuated S. typhimurium X4550/pYA3341-Cap. Vaccine tests in swine showed that the oral immunization with attenuated S. typhimurium X4550/pYA3341-Cap could elicit significantly higher Cap antibody titers in the treated swine than the control groups. Virus neutralization test showed that serum from the swine treated with attenuated S. typhimurium X4550/pYA3341-Cap had significant levels of neutralization activities. The swine lymphocyte proliferative responses indicated that attenuated S. typhimurium X4550/pYA3341-Cap could induce obvious cellular immune response. An in vivo challenge study showed the swine treated with attenuated S. typhimurium X4550/pYA3341-Cap had significantly lower PCV2-associated lesions and PCV2 viremia than the control groups. The results indicated that attenuated S. typhimurium X4550/pYA3341-Cap can be a potential vaccine against PCV2 infections.     

A fatty acid binding protein from Fasciola hepatica induced protection in C57/BL mice from challenge infection with Schistosoma bovis.

Three strains of mice (NMRI, C57/BL, BALB/c) were each immunized with a 12 kDa purified, native Fasciola hepatica fatty acid binding protein (Fh12) and challenged percutaneously with Schistosoma bovis cercariae. C57/BL mice immunized with Fh12 had significant reductions in S. bovis worm burden recoveries (96 and 87% reductions over controls in two separate experiments). When using NMRI or BALB/c mice, Fh12 alone or in Freund's adjuvant failed to induce significant protection against S. bovis. In C57/BL mice vaccinated against Fh 12, antibodies to the IgG2a isotype, but not to the IgG1 isotype, increased by 2 weeks after the second immunization and remained high through 8 weeks of S. bovis infection. Antibodies to S. bovis increased after 4 weeks of infection. Regarding cytokine production by spleen mononuclear cells, C57/BL mice vaccinated with Fh12 in adjuvant, and having the highest protective response against challenge infection with S. bovis, had an increase of IFN-gamma production with Concanavalin A but no increase of IL-4 in similarly stimulated cells. These results suggest that the protection obtained in this group of mice is mediated by a Th1 immune response.     

低功率氦氖激光加血卟啉衍生物对动物移植肿瘤的疗效观察

用血卟啉衍生物结合氦氖激光,治疗小鼠S180纤维肉瘤。其显效率高达74.51％,肿瘤消失率达64.71％;在治疗后第13、19天,肿瘤的平均重量和体积分别与治疗前比较,差异极显著(P<0.01);当激光剂量不变时,随着血卟啉衍生物用量增加,其疗效增加。     

A coagglutination assay with monoclonal antibodies for rapid laboratory identification of Mycoplasma synoviae.

An agglutinating monoclonal antibody (MAb S2) specific for a 55,000-molecular-weight surface protein of Mycoplasma synoviae was developed by fusion of spleen cells from immunized BALB/c mice with P3X63 Ag8.653 myeloma cells. Immunogold labeling experiments confirmed the cell surface location of the MAb-recognized antigen. MAb S2-coated Staphylococcus aureus was used in a rapid slide coagglutination assay. Eleven M. synoviae strains, including the type strain WVU 1853, coagglutinated with MAb-coated S. aureus, but five M. gallisepticum strains (PG31, S6, R, F, and A5969) did not.     

不同方法提取的蜂胶液中总黄酮含量的测定及其药理学研究

测定了不同方法提取的蜂胶液中黄酮类化合物的含量,并观察了黄酮类化合物含量较高的蜂胶醇提液和水提液对小鼠S180实体瘤模型、小鼠肉芽肿模型、大鼠急性关节炎模型和大鼠急性胸膜炎模型的影响.结果发现,在乙醇提取中,80%的乙醇提取液中的黄酮类化合物含量最高,达51.36mg/mL;特有水提的黄酮类化合物含量是普通水提含量的6.7倍.在相同剂量的情况下,蜂胶水提液和醇提液的抗炎和抗肿瘤效果差异不大甚至优于醇提.由于水提物刺激性小,易于调味,是一种极具开发潜力的提取方式.     

猪葡萄球菌感染裸鼠及BALB/c小鼠模型的研究

用一株本室分离鉴定能够引起猪渗出性皮炎的猪葡萄球菌(S.hyicus GZ1)经肌肉注射裸鼠和BALB/C小鼠,意图研究裸鼠和BALB/C小鼠对猪葡萄球菌的易感性及猪葡萄球菌对裸鼠和BALB/C小鼠的致病性。结果表明裸鼠和BALB/C小鼠均可被感染,并表现各自的临床症状。裸鼠在感染后2到3d背部和面部皮肤开始出现大量小的红色囊泡,4到5d后部分囊泡消失,部分形成结痂。一些裸鼠眼睛还会有较多脓性分泌物渗出,感染两周后相继死亡。BALB/C小鼠感染该菌后多表现急性临床症状,感染后2到3d就相继死亡,因此表现不出明显的眼观病变,只有很少一部分小鼠皮肤上会出现炎性渗出,导致该处皮肤脱落。研究表明:BALB/C小鼠比裸鼠对猪葡萄球菌更易感,裸鼠感染后产生清晰可见的临床症状,而BALB/C小鼠在感染后往往还来不及表现明显的临床症状就相继死亡。可见,猪葡萄球菌不仅对猪有致病性,对裸鼠和BALB/C小鼠也都表现出不同程度的致病性。     

Manipulation of the gut microbiota in C57BL/6 mice changes glucose tolerance without affecting weight development and gut mucosal immunity

Inflammatory diseases such as type 2 diabetes (T2D) in humans and mice are under the influence of the composition of the gut microbiota (GM). It was previously demonstrated that treating Lep(ob) mice with antibiotics improved glucose tolerance. However, wild type C57BL/6J mice may also exhibit plasma glucose intolerance reminiscent of human T2D. We hypothesized that antibiotic treatment in C57BL/6 mice would have an impact on glucose tolerance without affecting weight and gut immunology. When compared to mice treated with erythromycin or the controls, treatment for five weeks with ampicillin improved glucose tolerance without significantly affecting the weight or the number of gut mucosal regulatory T cells, tolerogenic dendritic cells or T helper cells type 1. 16S rRNA gene based denaturing gradient gel electrophoresis profiles clearly clustered according to treatment and showed that antibiotic treatment reduced GM diversity. It is concluded that antibiotic treatment changes glucose metabolism as well as the composition of the GM in C57BL/6 mice, and that this does not seem to be correlated to weight development in the mice.     

Isolation and characterization of temperature-sensitive mutants of Streptococcus suis: efficacy trial of the mutant vaccine in mice

A model of experimental Streptococcus suis infection was developed in young mice. Minimum lethal dose (MLD) values were calculated for four virulent serotypes (1/2, 1, 2, 3) of S. suis using this model. Temperature-sensitive (ts) mutants of S. suis serotypes 1/2 and 1-8 were isolated and characterized on the basis of their growth kinetics and reversion rates. Ts mutants of S. suis 1/2, 1, 2, and 3 were tested as vaccines against the virulent homologous and heterologous challenges in mice. The protection provided was evaluated by analyzing the clinical signs, death or survival. Homologous but not heterologous protection was noted in all mice vaccinated with the mutant strains. Ts mutants of S. suis 1/2 provided 100% protection against challenge by virulent strains of S. suis 1/2, 1, and 2.     

甘肃棘豆抗肿瘤活性成分的筛选

用甘肃棘豆提取物中乙酸乙酯极性部位、水相部位以及生物碱和非生物碱部位，对荷S180和H22肿瘤小鼠进行了体内抗肿瘤试验。结果，试验4个样品中，非生物碱部位没有抗肿瘤活性，乙酸乙酯部位和水相部位对S180和H22均具有一定的抑制作用，但作用都不及生物碱部位强；各试验组小鼠胸腺、脾和体重变化都不明显。结果表明，各样品试验用剂量对小鼠均无毒性反应。     

Cloning, expression and characterization of a cell wall surface protein, 6-phosphogluconate-dehydrogenase, of Streptococcus suis serotype 2

Streptococcus suis type 2 is a pathogen responsible for diverse diseases in both pigs and humans. In order to understand the pathogenesis of the S. suis type 2 infection, the gene encoding a cell surface protein, 6-phosphogluconate-dehydrogenase (6PGD) of S. suis type 2 was cloned and sequenced, and recombinant 6PGD protein (r6PGD) was produced in a prokaryotic expression system. Sequence analysis of the cloned 6 pdg gene showed 82% similarity with Streptococcus pneumoniae 6 pdg at the nucleic acid level. Western blotting using r6PGD-specific antiserum confirmed the cell surface location of the 6PGD protein of S. suis type 2. The role of 6PGD in S. suis type 2 pathogenesis as an adhesin and its immunogenicity in mice was further investigated. The results showed that the recombinant protein interfered with the adhesion of S. suis type 2 to Hep2 and HeLa cells by 72% and 66%, respectively. Immunization of CD-1 mice with r6PGD increased the protective efficacy by 80% following intraperitoneal administration of a lethal dose of S. suis type 2. Immunization of CD-1 mice with r6PGD elicited a significant protective immune response, which demonstrated the importance of 6PGD to bacterial pathogenesis. Identification and characterization of the role of S. suis type 2 6PGD in adhesion and immunogenicity will allow us to use this protein to develop new antimicrobial therapies and/or vaccines.     

Experimental infections of mice and pigs with Streptococcus suis type 2.

Five inbred strains of mice were tested for their susceptibility to Streptococcus suis type 2 including the type strain, two isolates from meningitis in pigs and two isolates from tonsils of clinically healthy pigs. C57BL/6, ICR and ddY strain mice showed lower susceptibility to all strains of S. suis type 2 than BALB/c and SS strain mice. The type strain and the isolates from diseased pigs produced septicaemia and meningitis in BALB/c and SS mice inoculated with 10(8) colony forming unit of the bacteria and 60 to 100% of these infected mice died. On the other hand, mice inoculated with the isolates from healthy pigs showed mild clinical signs but none of them died. In BALB/c mice which died or developed nervous signs, the purulent meningo-encephalitis, myocarditis, ophthalmitis, labyrinthitis and otitis media were observed. S. suis type 2 antigen was demonstrated in these lesions by immunoperoxidase staining using rabbit S. suis type 2 antiserum. These results were similar to those in the experimentally infected pigs with these virulent and avirulent strains against mice. These results indicate that BALB/c and SS strains of mice are useful as an experimental model of S. suis type 2 infections in pigs, and that there are virulent and avirulent strains against mice and pigs among the strains of S. suis type 2.     

Cell-cycle-dependent Colonization of Mouse Spermatogonial Stem Cells After Transplantation into Seminiferous Tubules

Spermatogonial stem cells (SSCs) migrate to the niche upon introduction into the seminiferous tubules of the testis of infertile animals. However, only 5–10% of the transplanted cells colonize recipient testes. In this study, we analyzed the impact of cell cycle on spermatogonial transplantation. We used fluorescent ubiquitination-based cell cycle indicator transgenic mice to examine the influence of cell cycle on SSC activity of mouse germline stem (GS) cells, a population of cultured spermatogonia enriched for SSCs. GS cells in the G1 phase are more efficient than those in the S/G2-M phase in colonizing the seminiferous tubules of adult mice. Cells in the G1 phase not only showed higher expression levels of GFRA1, a component of the GDNF self-renewal factor receptor, but also adhered more efficiently to laminin-coated plates. Furthermore, this cell cycle-dependency was not observed when cells were transplanted into immature pup recipients, which do not have the blood-testis barrier (BTB) between Sertoli cells, suggesting that cells in the G1 phase may passage through the BTB more readily than cells in the S/G2-M phase. Thus cell cycle status is an important factor in regulating SSC migration to the niche.     

Early sequential ultrastructural renal alterations induced by 2-bromoethylamine hydrobromide in the Swiss ICR mouse.

Thirty-two male Swiss ICR mice were injected intraperitoneally with 300 mg 2-bromoethylamine hydrobromide/kg body weight, anesthetized, and perfused with glutaraldehyde-paraformaldehyde solution at 5, 15, 30, 60, 90, 120, 150, and 180 minutes after treatment. Eight control mice were injected intraperitoneally with sterile diluent, and one was perfused at each of the same time periods as the treated mice. Proximal tubule epithelial alterations progressed over time from increased secondary lysosome and myeloid body formation to cellular and mitochondrial swelling and eventually cell necrosis. The glomerular, peritubular, and vasa recta capillaries had endothelial cell swelling and desquamation and platelet aggregation. Bromoethylamine nephrotoxicosis in the male Swiss ICR mouse is an ischemic necrosis of the proximal tubules and papilla initiated by endothelial cell damage and makes an excellent model of chemically induced damage to endothelial cells and tubular necrosis.