Recombinant human hepatitis B vaccine initiating alopecia areata: testing the hypothesis using the C3H/HeJ mouse model

Untoward effects of human vaccines suggest that recombinant hepatitis B vaccine may induce alopecia areata (AA) in some patients. Similar untoward immunological effects may also account for AA-like diseases in domestic species. In this study, the C3H/HeJ spontaneous adult onset AA mouse model was used to test the role, if any, of recombinant hepatitis B vaccine on the initiation or activation of AA. Initial experiments demonstrated no effect on induction of AA in young adult female C3H/HeJ mice ( P  = 0.5689). By contrast, older females, those at the age when AA first begins to appear in this strain, had a significant increase ( P  = 0.0264) in the time of onset of AA, suggesting that the vaccine may initiate disease in mice predisposed to AA. However, larger vaccine trials, which included diphtheria and tetanus toxoids as additional controls, did not support these initial result findings and suggest that AA associated with vaccination may be within the normal background levels of the given population.     

Experimental infection of C3H/HeJ mice with the NY18 isolate of Anaplasma phagocytophilum

Human granulocytic anaplasmosis (HGA), an emerging disease of public health concern in many areas of the world, is caused by Anaplasma phagocytophilum. Small animal models of A phagocytophilum in laboratory mice have been developed and used to study the pathogenesis of HGA. In this study, we characterized the pathologic changes in acute infection of C3H/HeJ mice experimentally infected with the NY18 isolate of A phagocytophilum. Although no clinical signs were noted, acute infection was associated with gross splenomegaly, microscopic inflammatory lesions in the lung and liver, hyperplastic lesions on the spleen, and clinical pathology abnormalities including neutropenia and monocytosis. This study emphasizes the use of well-defined animal models as a valuable tool for the study of A phagocytophilum infections.     

Humoral antibody and lymphocyte blastogenesis responses in BALE/c, C3H/HeJ and AKR/N mice following Ehrlichia risticii infection

Three strains of mice, two susceptible to Ehrlichia risticii induced disease (balb/c and C3H/HeJ) and one resistant (akr/n), were evaluated for the development of Immoral and cell mediated immune responses following infection with E risticii. The production of serum antibody was determined by indirect fluorescent antibody testing of sera from mice of each strain challenged with one of three different dose levels of E risticii. Antibody was assayed on days 7, 9, 12, 15 and 20 after inoculation. Cell mediated immune responses were evaluated by measuring the blastogenesis response of spleen cells from E risticii infected mice 28 days after inoculation. All three strains of mice at the high challenge level responded with the production of antibody by day 9 after inoculation. Overall, the antibody response occurred earlier and was of greater magnitude in the susceptible balb/c and C3H/HeJ strains. A marked blastogenesis response occurred in splenocytes from E risticii infected mice of all three strains upon re-exposure to ehrlichial antigen. The findings of this study indicate that susceptibility to E risticii induced disease was not the result of deficient or delayed humoral immune responses and that E risticii infection induced the development of strong cell mediated immunity.     

Infection of normal C3H/HeN mice with Taenia saginata asiatica oncospheres

Normal C3H/HeN female mice were used to develop an animal model of Taenia saginata asiatica oncosphere infection. The host cellular immune response in this model was analyzed by a cytokine enzyme-linked immunosorbent assay (cytokine ELISA) and flow cytometry. Tumor-like cysts containing cysticerci were recovered from the inoculation sites of female mice 7 weeks postinfection with the T. saginata asiatica oncospheres. A sharp increase and sustained elevation in the ability of spleen cells to produce interferon-γ and interleukin (IL)-2 revealed that cellular immunity played an important role during the infection. An immediate increase in the levels of IL-6 at 1 week postinfection indicated the induction of a local acute inflammatory response. However, no significant change in the levels of IL-10 indicated that Th2 cells were not involved in this immune response. The patterns of cell distribution revealed by flow cytometry also supported the same finding. These results suggested that Th1 cells played a major role in the immune response in C3H/HeN mice during the early stages of the oncosphere infection and that the Th2 response was not induced during the stage of cysticercus formation.     

Effect of monoenergetic neutron irradiation on the postnatal development of the cochlea in C3H/HeN mice

To investigate the toxic effect of neutrons at energies of approximately 1MeV on the ear, we exposed 7-day-old mice to 1.0 Gy of monoenergetic neutrons (1.026 MeV) or (137)Cs gamma rays, and assessed subsequent morphological changes in the inner ear by light and scanning electron microscopy. Monoenergetic neutrons, but not gamma rays, caused acute changes in the ear. The epithelium of the greater epithelial ridge in the organ of Corti had disappeared by 72 hr post-irradiation, as a result of epithelial apoptosis observed 6 hr post-irradiation. Radiation could induce apoptotic cell death of the epithelium of the greater epithelial ridge at 3 or 4 days of age. Protruding structures were detected on the surface of the hair cells by 72 hr post-irradiation. The neutron-irradiation also caused the apoptotic cell death of epithelial cells at the nasal conchae, and subsequent acute otitis media continued until 10 weeks of age.     

High mortality with severe dystrophic cardiac calcinosis in C3H/OUJ mice fed high fat purified diets

Severe degenerative myocardial disease occurred in female C3H/OUJ mice fed purified diets for 36 weeks; the diet contained 5% or 20% fat as non-hydrogenated soybean oil. Deaths of lactating females of this group (17/35 high fat diet and 7/35 low fat diet animals) were due to sudden cardiovascular collapse. Cardiomegaly with marked atrial and ventricular myocardial mineralization was seen at necropsy. Histologically, the random, myopathic foci were characterized by severe myocardial degeneration, mineralization, and fibrosis. Mural thrombosis, pulmonary arteriosclerosis, and mild myocardial inflammatory cell infiltrates were also present. Pathological changes were similar to those of dystrophic cardiac calcinosis, an incidental necropsy finding in certain mouse strains.     

Differences in the kinetics of T cell accumulations in C3H/HeN (Bcg-resistant) and C57BL/6 (Bcg-susceptible) mice infected with Mycobacterium paratuberculosis

The accumulation of various T cell subsets in Bcg-susceptible (C57BL/6) and -resistant (C3H/HeN) strains of mice were compared following an intraperitoneal infection with Mycobacterium paratuberculosis. Groups of mice from both strains were killed at 3, 5, 10, 15, 30, and 150 days after infection and lymphocytes were harvested from the peritoneal exudate cells (PEC), spleen, intestinal epithelial lymphocytes (IEL), lamina propria lymphocytes (LPL), Peyer's patches, and mesenteric lymph node (MLN) and labelled with monoclonal antibodies to CD3, CD4, CD8, γδ TCR, CD25, and CD44 for flow cytometric analysis. Uninfected C3H/HeN mice had higher proportions of CD4+ cells in the spleen, MLN, LPL, IEL and Peyer's patches, while uninfected C57BL/6 mice had higher proportions of CD8+ and/or γδ T cells. Significant increases in accumulation of CD8+ and γδ T cells were detected in the peritoneum and other tissues in both strains of mice after infection. Higher CD4/CD8 ratios were observed in most lymphoid tissues of C3H/HeN mice, while increased proportions of CD8+ and/or γδ T cells were present in C57BL/6 mice. These results indicate that significant differences in T cell profiles exist between these two strains of mice, both inherently and in response to infection with M. paratuberculosis. Innately lower levels of CD4+ cells and/or higher percentages of CD8+ and γδ T cells may play a role in the increased susceptibility of C57BL/6 mice to infection with M. paratuberculosis.     

Prime-boost immunization with HA/C3d DNA followed by a recombinant pseudorabies virus boost enhanced protective immunity against H3N2 swine influenza virus in mice

DNA and recombinant virus vaccines against swine influenza virus (SIV) have been pursued with promising results, but induce poor immunogenicity. This study evaluated the effects of a vaccine regimen in mice including priming with three DNA vaccines expressing soluble HA (sHA), complete HA (tmHA), or sHA fused with three copies murine C3d (sHA-mC3d3) and boosting with recombinant pseudorabies virus expressing HA (rPRV-HA). Immune responses were monitored by ELISA, HI assays, and virus neutralization. Protective efficacy was evaluated by virus isolation from lungs, distribution in tissues, and pathology following challenge with H3N2 SIV. Priming with sHA-mC3d3 and boosting with rPRV-HA induced higher levels of HA-specific antibodies and yielded the most effective protection. This finding implied that priming with a DNA vaccine expressing C3d fused with antigen and boosting with a recombinant vector vaccine is an effective way to induce protective humoral immunity and prevent some infectious diseases.     

Morphometric analysis of enteric lesions in C3H/HeN mice inoculated with Serpulina hyodysenteriae serotypes 2 and 4 with or without oral streptomycin pretreatment.

The segmental distribution and sequential progression and the role of the indigenous bacterial flora in the development of enteric lesions associated with Serpulina hyodysenteriae infection in laboratory mice have not been defined. We examined the distribution and sequential morphometric changes in the large intestine of mice orally inoculated with S. hyodysenteriae serotypes 2 and 4. To determine the role of colonization resistance conferred by the indigenous bacterial flora, 40 female C3H/HeN mice were administered water alone or water containing 5 mg/mL streptomycin sulfate ad libitum for seven days prior to orogastric inoculation either with S. hyodysenteriae or sterile trypticase soy broth (TSB). Clinical signs were monitored daily and three mice per group were necropsied on postinoculation days (PID) 7 and 14 for pathological assessment of the cecum, proximal colon, transverse colon, and descending colon, and bacteriological culture of the cecum for S. hyodysenteriae. Weekly pooled fecal samples were collected from each group for determination of total numbers of anaerobe bacteria. Gross examination revealed soft fecal pellets on PID 7 and 14 and catarrhal typhlitis on PID 14, irrespective of streptomycin pretreatment. The recovery rates of S. hyodysenteriae from the ceca of serotype 2- and serotype 4-inoculated mice was 100 and 91.7%, respectively. Statistically significant differences in morphometric changes between TSB- and S. hyodysenteriae-inoculated mice were present on PID 7 and 14 and were restricted to the cecum. Although oral administration of streptomycin for seven days prior to S. hyodysenteriae inoculation resulted in a significant reduction in the numbers of fecal anaerobes, it did not affect the colonization, distribution, severity, or progression of cecal lesions.     

过敏毒素C5a和C3a在微小隐孢子虫感染中的表达分析

为明确过敏毒素C5a和C3a在小鼠感染和清除隐孢子虫(Cryptosporidium)感染过程中的作用,本研究以微小隐孢子虫(C.parvum)为研究对象,以BALB/c小鼠为感染模型,采用实时荧光定量PCR分析了C.parvum感染小鼠的小肠相关淋巴组织和脾脏中过敏毒素C3a、C5a及其相应受体(C3aR和C5aR)的mRNA水平。结果显示,在脾脏和小肠中,不仅过敏毒素C5a和C3a在感染后出现了显著上调表达,其受体C5aR和C3aR的表达水平在感染后亦显著上调表达(P<0.05),并呈现动态变化。这些研究结果提示C5a/C5aR和C3a/C3aR信号参与了宿主抗微小隐孢子虫感染的过程。     

Comparison of the effectiveness of high and low LET radiations for the proportion of survivals with liver tumors at every age in (C57BL/6N x C3H/HeN) F1 mice

To investigate the late effects of neutrons at the energy below 1 MeV on the liver carcinogenesis as a function of age, one-week old mice were exposed to 1.0 Gy monoenergetic neutrons (0.317, 0.525 and 1.026 MeV) or 137Cs gamma rays. Survival and carcinogenesis were examined by 18 months of age. Following radiation, tumor incidences in liver, Harderian gland, lung, ovary and pituitary gland were compared. The proportion of the lifespan with liver tumors exposed to neutrons to that exposed to gamma rays was calculated as a function of age. Survival rates among the three groups exposed to neutrons of different energies were not significantly different from one another but shorter than those treated with gamma rays for both sexes. With regard to liver tumor incidence evaluated at 18 months of age, the effectiveness of neutrons to gamma rays was 2.54 for females, and 2.08 for males by the factor. Levels of estrogen in the serum were similar between mice bearing liver tumors and those devoid of tumors. In conclusion, all three energies of neutrons induced similar effectiveness with respect to liver carcinogenicity. Proportions of the lifespan with liver tumors of neutron-exposed to gamma-exposed were shorter in females than males along with ages over 12 months. To obtain this factor at every age contributed for the evaluation of the biological effectiveness of radiations with the parameter of tumor incidence and latency simultaneously.     

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。     

Two Cases of Metastatic Parathyroid Carcinoma in Male C3H Mice Following Irradiation

White nodules were observed in the thyroid in two male C3H mice (at 99 and 122 weeks of age) exposed to fast neutrons at the age of 8 weeks. Histopathologically, in both cases, tumors were developed in the region corresponding to the parathyroid gland, and the tumor cells were arranged in a solid sheet or nest-like structures. Necrosis, cell debris and/or hemorrhage were sometimes seen in the center of the tumor structures. Tumor cells were small and uniform with scanty cytoplasm, cell margins were indistinct, and basally located tumor cells were aligned along the vascular stroma. Mitotic figures were frequently observed. Metastasis to the renal cortex was observed in both cases. These cases were diagnosed as parathyroid carcinoma. A parathyroid tumor is an extremely rare endocrine tumor in mice, regardless of whether the tumor is spontaneous or experimentally induced. These cases may have been induced by neutron-exposure; however, how radiation induces parathyroid carcinoma in mice is not clear.     

对C57BL／6小鼠超排效果的研究

本文以近交系C57BL／6小鼠为实验对象．研究注射不同剂量的PMSG和hCG对小鼠超排效果的影响。取C57BL／6小鼠各30只,按照注射剂量不同分为A、B、C三组,每组10只,A组注射PMSG2．5IU,HCG2．5IU,B组注射PMSG5．0IU,HCG5．0IU,C组注射PMSG7．5IU,HCG7．5IU。每只小鼠腹腔注射PMSG,间隔48h后分别注射HCG进行超数排卵,再与性成熟同系公鼠合笼,次日早上检查阴道栓．有栓雌鼠用颈椎脱臼法处死。在实体显微镜下由输卵管膨大部冲卵．收集卵母细胞置于盛有M2培养液的表面皿中检查计数．分析超排效果。结果表明。C57BU6小鼠B组的平均取卵数极显著高于A组的平均取卵数（P〈0．05）;B组的平均取卵数显著高于C组的平均取卵数（P〈0．05）;C组与A组的平均取卵数差异不显著（P〉0．05）。     

Nine cases of granular cell tumors in B6C3F1 mice

The histological characteristics of 9 cases of granular cell tumors (GCTs) observed in B6C3F1 mice were examined to determine their cellular origin. Seven of the 9 cases were found in the uterus and other 2 cases were in the subcutaneous tissue. Tumor cells had abundant granules in the cytoplasm which were stained with PAS and were resistant to diastase treatment. Ultrastructurally, the granules were identified as lysosomes. The cell surface had cytoplasmic processus showing interdigitation with adjacent cells. A character feature of the tumor cells was the presence of a desmosome-like structure on their cell surface but no basal lamina was demonstrated. Although GCTs have been considered to be derived from Schwann cells on the basis of their ultrastructural features and S-100 protein-immunopositive findings, the absence of basal lamina in the present cases may raise a controversy as to their origin.     

FGF20及其受体FGFR2和FGFR3在小鼠毛囊形成期的定位及表达

旨在研究成纤维细胞生长因子(FGF20)及受体FGFR2和FGFR3在小鼠胚胎期毛囊形成期的表达情况,了解FGF20及受体FGFR2和FG-FR3在小鼠毛囊形成期的作用。采用实时荧光定量PCR、Western blot和免疫组织化学技术检测胚胎期13 d (E13)至18 d (E18)小鼠背部皮肤中FGF20、FGFR2和FGFR3 mRNA及蛋白的差异表达。免疫组化结果显示,FGF20蛋白和FGFR3蛋白E13在表层细胞微弱表达,E14主要表达在表层细胞,且在基底层细胞也有微弱表达,E15主要表达在基板和表层细胞,E16强表达在毛钉与表层细胞,E17和E18主要表达在表层细胞和未成熟的毛囊;FGFR2蛋白在表层细胞、基底层细胞、基板、毛钉、未成熟的毛囊等处均有表达,且E18强表达在表层细胞和未成熟毛囊;RT-PCR及Western blot数据分析结果显示:FGF20 mRNA及蛋白在E16相对表达量最高;FGFR2 mRNA及蛋白在E13相对表达量最低,E18相对表达量最高;FGFR3 mRNA及蛋白表达趋势和FGF20相似。研究结果提示,在毛囊形成期,FGF20可能通过与FGFR3结合从而参与毛囊的诱导和激活,促进毛囊形成并决定其生长方向。FGFR2可能在毛囊形成过程调控表层细胞增殖,促进毛囊形成,诱导毛囊进入第一生长周期。     

JMJD1C对牛卵丘细胞H3K9me1、H3K9me2的去甲基化作用

KDM3家族成员中KDM3A、KDM3B能够特异性去除H3K9me1与H3K9me2的甲基化修饰,而JMJD1C作为KDM3家族成员之一,与KDM3A、KDM3B即JMJD1A、JMJD1B含有相似的Jmj C结构域,但JMJD1C是否也具有对H3K9的催化作用或活性一直不明确。本研究利用RNAi方法抑制JMJD1C的mRNA表达水平,随后通过免疫荧光方法检测H3K9me1与H3K9me2的甲基化程度。结果表明,3T3-L1细胞和卵丘细胞中均存在JMJD1C的表达,并且存在H3K9me1与H3K9me2的甲基化修饰。通过siRNA对牛卵丘细胞中JMJD1C mRNA的表达进行成功抑制后,转染组JMJD1C的表达量明显下降,但免疫荧光结果显示H3K9me1、H3K9me2表达量无明显变化,即JMJD1C在mRNA水平上对去除H3K9me1与H3K9me2的甲基化修饰并无明显作用。     

Pathological changes in Streptobacillus moniliformis infection of C57bl/6J mice

An epizootic disease caused by Streptobacillus moniliformis occurred in C57BL/6J mice. Pathological lesions included abscessation of lymph nodes and chronic polyarthritis and osteomyelitis. Histological features of the disease are described. The most important differential diagnosis, infectious ectromelia of mice, is discussed.     

C57BL/6J小鼠亚慢性镉中毒模型的建立

为了建立小鼠亚慢性镉中毒模型,本研究将20只C57BL/6J雄性小鼠随机分为4个剂量组和溶剂对照组,每组隔天分别腹腔注射含0.25、0.5、1和2 mg/kg剂量的氯化镉溶液和等量去离子水(溶剂),共染毒4周,观察不同剂量的镉离子对雄性小鼠肝脏、肾脏、睾丸的损伤情况.结果各处理组小鼠体质量增长要比对照组慢,脏器系数与对照组相比也有显著差异,处理组小鼠肝脏、肾脏、睾丸中镉含量显著高于对照组.病理组织切片结果表明,各处理组中小鼠的肝脏、肾脏、睾丸组织均出现不同程度的损伤.氯化镉可以显著地抑制小鼠体质量增长,并对小鼠的脏器系数产生影响,腹腔注射后在肝脏、肾脏、睾丸组织中产生蓄积,并对其造成一定损伤.隔天腹腔注射2 mg/kg剂量的氯化镉,4周可建立较理想的小鼠镉中毒模型.