黑老虎果实提取物对小鼠腹泻模型的干预作用

为了研究黑老虎果实提取物的抗腹泻作用,从而为临床应用提供实验依据。实验首先对黑老虎果实提取物中木脂素、三萜及黄酮含量进行测定,然后对ICR小鼠进行细菌性腹泻、药物性腹泻和小肠炭末推进实验。根据实验要求,将细菌性腹泻模型小鼠和药物性腹泻模型小鼠灌胃不同药物,连续7天,除正常组外,其余小鼠于末次给药30min后分别腹腔注射大肠埃希菌溶液和灌胃番泻叶提取液,观察并记录各组小鼠腹泻情况;小肠炭末推进实验于小鼠末次给药30min后灌胃给予5%炭末,计算小肠炭末推进率 ;结果表明,黑老虎果实提取物中木脂素类、三萜类及黄酮类化合物的含量分别为44.48%、4.57%、14.02%黑老虎果实提取物各剂量组对细菌性小鼠均有较好的预防效果,其中高剂量组 （200mg/kg）最佳,与模型组呈显著差异（P＜0.01）,效果亦优于阳性药物组;提取物各剂量组均能减少番泻叶腹泻小鼠稀便数量,且与模型组呈显著差异,但对潜伏时间没影响;提取物对小鼠小肠炭末推进无明显影响 由此可知,黑老虎果实提取物对细菌性腹泻小鼠具有良好的抗腹泻作用,对药物性致腹泻作用较弱。     

大麦苗营养及其对白鼠健康的影响

为了对大麦苗的开发利用提供必要的理论依据,以闽诱3号为试验材料,对分蘖盛期的大麦苗进行营养分析。结果表明,大麦苗中含有丰富的蛋白质(31.26％)、氨基酸(其中赖氨酸达到了1．1445％)、矿物质(包括Fe、K、Cu、Mg、Ca、P、Mn、Zn等8种元素,以Fe含量最高．达到8580．0μg／m1)、维生素(以维生素C含量最高,达到65．570μg／m1)和SOD酶(0．555μg／m1)等。用大麦苗制品喂养白鼠发现有以下几方面明显作用：①瘦身、抗肥胖;②降低血液中有害物质脂质过氧化物、甘油三脂和胆固醇的含量;③提高血液中有益物质高密度脂蛋白和SOD酶的含量;④减少酒精对胃粘膜的损伤,保护胃粘膜;⑤抗疲劳,增强体力。这些结果表明,大麦苗营养对生命肌体有维护和保健功效。     

Interleukin 17 (IL-17) manipulates mouse bone marrow- derived neutrophils in response to acute lung inflammation

Interleukin 17 (IL-17) mediates neutrophil migration to the lungs during acute inflammation, potentially leading to lung tissue damage. In the present study, we evaluated whether IL-17 could facilitate certain neutrophil functions in a mouse model. Mice were divided into four groups and intranasally challenged with PBS (1 = Control), Influenza A (H1N1) and Klebsiella pneumoniae (2 = Mix), Influenza A alone (3 = Flu), or K. pneumoniae (4 = KP) alone. Bone marrow, BAL cells, and lung specimens were collected seven days post-challenge for analysis. Mice in the Flu group showed the highest mortality rate. Neutrophils were the prominent cell type in BAL from Mix and KP, whereas lymphocytes were most numerous in Flu. Lesions in the lungs revealed considerably damage in the Mix, Flu, and KP groups. Isolated bone marrow-derived neutrophils were in vitro primed with mouse recombinant IL-17A protein (rIL-17A) followed by various functional assays. The reactive oxygen species (ROS) levels in rIL-17A primed cells showed significant elevations in all groups. Phagocytosis and bacterial destruction showed no significant difference between (+) or (−) rIL-17A groups. The formation of neutrophil extracellular traps (NETs) in rIL-17A-primed neutrophils showed elevated NET production. We next monitored expressions of genes in neutrophils. IL-17A mRNA expression was significantly increased in Mix and Flu; IL-1β mRNA only significantly increased in Flu, and IL-17RA showed constitutive expressions in all groups. In summary, neutrophils may cause tissue damage during lung inflammation through specific functions influenced by IL-17.     

Direct embryotoxicity of chromium (III) exposure during preimplantation development

Chromium in its trivalent form (chromium (III)) is an essential component of a balanced diet, and its deficiency disturbs glucose and lipid metabolism in humans and animals. The prevailing view is that chromium (III) is notably less toxic than chromium (VI), which is genotoxic and carcinogenic. Thus, the biotransformation of environmental chromium (VI) to chromium (III) is a promising and environmentally friendly detoxification method. However, increasing evidence suggests that chromium (III) induces considerable cytotoxicity. However, the toxicity of chromium (III) to early embryos remains largely unknown. In the present study, we used in vitro fertilization (IVF) to produce mouse embryos and identified the direct embryotoxicity of chromium (III). On exposure to high concentrations of CrCl₃, blastocyst formation almost completely failed and a large proportion of embryos were arrested at the 2- to 4-cell stage. At low concentrations of CrCl₃, IVF embryos showed a significant decrease in blastocyst formation, reduced total cell numbers, aberrant lineage differentiation, increased oxidative stress, and apoptosis. We also found that chromium (III) exposure during the preimplantation stage, even at low concentrations, led to impaired post-implantation development. Thus, our study substantiates the direct embryotoxicity of chromium (III) during preimplantation development and prolonged impairment of development potential. The results further highlight the potential adverse effects of chromium (III) on public reproductive health with respect to increased environmental enrichment of and dietary supplementation with chromium (III) complexes.     

随机限食对小鼠代谢产热的影响

[目的]研究随机限食条件下KM小鼠的代谢产热,深入理解能量代谢和产热在动物适应限食应激中的作用和意义。[方法]成年KM小鼠随机限食4 d周/,共驯化4周,采用封闭式流体压力呼吸计测定基础代谢（BMR）和非颤抖性产热（NST）。[结果]对照组动物体重增长了15.1%,而限食组降低了5.3%,限食组显著低于对照组（P〈0.01） 随机限制显著增加了摄食量,摄食量最大值为（11.94±0.52）g/d,比对照组高132.8%（P〈0.001） 限食使BMR（P〈0.05）和NST（P〈0.05）显著降低 随机限食使胴体重显著降低（P〈0.01）,但未影响脂肪重量和含量（P〉0.05）。[结论]动物能通过能量摄入和支出的权衡策略适应随机限食的应激环境,代谢和产热的适应性调节在能量代谢的权衡策略中发挥重要作用。     

Lateral canthoplasty for facial nerve paralysis and exposure keratitis in a mouse

An 11-month old entire male mouse was presented with mucopurulent discharge and corneal scarring of the left eye. This mouse previously had a history of ear mites that responded to therapy, and had undergone surgery to remove a large discharging mass from the left side of the neck. The eye problem was noted prior to surgery, but after the ear mite infection. Examination revealed absence of a palpebral reflex in the left eye, and no spontaneous movements of the left ear or lip and whiskers. Examination of the left eye revealed extensive corneal vascularisation and pigmentation. Intraocular structures were not visible. A diagnosis of facial nerve paralysis and secondary exposure keratitis was made. Surgery was performed to close the lateral canthus and reduce corneal exposure. Following surgery the eye discharge ceased and corneal vascularisation resolved, however corneal pigmentation persisted.     

High-efficient genetic transfection of CD41+,UT7, U937 and MDA-MB-435 cells with a recombined murine stem cell retroviral vector

AIM: Gene transduction with a recombined murine stem cell retroviral vector has been investigated to find an effective way of gene transduction and to offer theory and experimental basis for the recombined murine stem cell retroviral vector used for gene transduction. METHODS: 1. Construction of retrovirus vector: EC1-4 gene (repeats 1-4 of cadher in-5 extracellular domain) and mutant (Ser 222A) MEK1 gene were cloned into retrovirus vector pMSCV after cut by Bgl Ⅱ and EcoR 1 restriction endonuclease. 2. Obtaining CD41⁺ cells and cell culture: Cells expressing CD34⁺ from cord blood were isolated. The inducement of cells expressing CD41 from CD34⁺ cells was performed by using TPO and cells CD41⁺ were selected by FACS. NIH 3T3 cells were cultured in high sugar DMEM medium and U937 in RPMI 1640 medium. UT7 cells which is cytokine-dependent cell line were grown in Iscove's modified Dulbeco's medium supplemented by GM-CSF. 3. Determination of viral titers: Retroviral vectors were transferred to packaging cell line 293. Retroviral containing supenatant was collected after transfection. The viral titers was tested on infection of NIH 3T3 cells by FACS analysis. 4. Western blot: Transduced CD41⁺, UT7, U937 and MDA-MB-435 cells were analyzed by western blot to examine expression of transduced genes. RESULTS: A packaging cell line 293 produces high-titer MEK1 pMSCV retroviruses (3.1×10⁷) and EC1-4 pMSCV retroviruses (1.0×10⁸). With 8-folds dilution retroviruses, 60.73% GFP positive cells have been obtained in MEK1 pMSCV transduced UT7 cells, 72.56% in U937 cells and 30.57% in CD41⁺ cells, respectively. GFP positive cells have reached up 97.54 % in EC1-4 pMSCV transducted MDA-MB-435 cells. Phosphorylated MEK1 has been decreased in experiment group when TPO has stimulated CD41⁺ and UT7 cells or serum has stimulated U973 cells. This indicates that is a dominant negative effect of mutation MEK gene. EC1-4 gene transduced MDA-MB-435 cells have expressed EC1-4. CONCLUSION:The recombined murine stem cell retrovirus can effectively mediate gene transduction of CD41⁺,UT7,U937 and MDA-MB-435 cells,and transduced genes can be stably expressed.     

Sox2 induce mouse embryonic fibroblasts direct reprogramming into neural stem cells

AIM: To study the direct reprogramming method of mouse embryonic fibroblasts (MEFs) converted into induced neural stem cells (iNSCs). METHODS: Sox2-infected MEFs were cultured in NSCs culture medium for 10 d. Subsequently, repeated suspension and adherent culture were performed for 3 times for the purification of iNSCs. The iNSCs were cultured in suspension medium. Real-time PCR was used to detect the expression of neural stem cell marker genes and pluripotent marker gene. In vivo, iNSCs were microinjected into the mouse cerebral cortex. Immunofluorescence was performed to detect the expression of neural stem cell, neuron, oligodendrocyte and astrocyte markers in vitro and vivo. RESULTS: A variety of neural stem cell marker gene expression was significantly increased in iNSCs detected by real-time PCR. Immunofluorescence confirmed that iNSCs expressed nestin and differentiated into neurons, oligodendrocytes and astrocytes in vitro and vivo. CONCLUSION: Sox2 is sufficient to trigger the direct reprogramming from MEFs to iNSCs. iNSCs have the ability of self-renew and 3 differentiation potentials in vivo and vitro. iNSCs are the suitable seed cells of SCI.     

Role of polymorphonuclear neutrophils (PMNs) and NK cells in the protection conferred by different vaccines against Chlamydophila abortus infection

Ovine enzootic abortion (OEA) is caused by Chlamydophila abortus, an intracellular bacterium which acts by infecting the placenta, causing abortion in the last term of gestation. The main prevention strategy against OEA is the vaccination of flocks. An effective vaccine against C. abortus must induce a Th1-like specific immune response, which is characterized by the early production of IFN-gamma and the activation of CD8(+)T cells. Moreover, vaccine effectiveness could be modulated by the functioning of the innate immunity. The purpose of this study was to ascertain how polymorphonuclear neutrophils (PMNs) and NK cells might influence vaccine-induced protection. The live attenuated 1B vaccine and two inactivated experimental vaccines, adjuvated with aluminium hydroxide (AH) or QS-21 (QS), were used in PMN-depleted or NK cell-depleted mice. For PMN depletion, RB6-8C5 monoclonal antibody, which recognizes GR1(+) receptors (Robben, P.M., LaRegina, M., Kuziel, W.A., Sibley, L.D. 2005. Recruitment of Gr-1(+) monocytes is essential for control of acute toxoplasmosis. The Journal of Experimental Medicine 201, 1761-1769.) was used, while for NK cell-depletion the anti-asialo GM1 polyclonal antibody was used. The depletion of PMNs caused 100% mortality in non-vaccinated mice (NV) and 60% mortality in the AH-vaccinated mice by day 10 p.i., while both groups showed a significant increase in their bacterial burden in the liver by day 4 p.i. The depletion of NK cells caused mortality only in the NV group (50% by day 10 p.i.), although this group and the 1B vaccinated mice showed an increased bacterial burden in the liver at day 4 p.i. Our results suggest that the importance of PMNs in inactivated vaccines depends on the adjuvant chosen. The results also demonstrated that the importance of NK cells is greater in live vaccines than in inactivated vaccines.