大鼠亚慢性试验随机表的生成与应用

[目的]研究大鼠亚慢性试验随机分组方法。[方法]用Excel软件对每只称重后的大鼠产生一个随机数,然后按随机数从小到大排序,以随机数的大小顺序将每只大鼠分入随机组中,并对体重均值作F检验。[结果]用Excel软件工具菜单中"录制宏"的功能,建立亚慢性试验随机分组模板文件。应用快捷键,快速完成随机分组及F检验。[结论]利用Excel模板文件方法可以快捷将实验动物随机分组。     

利用计算机进行区组随机分组的一种新方法及其应用

计算机产生的一组随机数是均衡分布于 0 - 1之间 ,利用该原始随机数进行区组随机分组 ,通常的方法有取整数法、取余数法 ,但这些方法均无法避免因简单随机产生的不平衡 ,特别是区组内 ,一般人们往往对原始随机数进行人为干预 ,例如删除、重新产生、人为指定等 ,以获得区组内平衡。本文描述了一种新方法———排序法 ,该法不需要对随机数进行上述人为干预 ,在简单处理后通过两次排序即可实现区组化随机分组。只要有产生随机数和能进行排序的计算机软件 ,均可运用本法实现区组化随机分组 ,本文例子采用的是JMP软件。临床试验设计中往往需进…     

农药毒理学实验随机分组快捷方法的建立

目的探讨农药毒理学实验随机分组的快捷方法。方法依据农药登记毒理学试验方法GB 15670-1995的要求,建立农药毒理学各试验的随机分组模板;并在Excel软件建立随机分组菜单,将各随机分组模板嵌入菜单。结果只要打开Excel软件,就可显示随机分组菜单,点击指定菜单,更新原始体重后,立即可完成相关试验的随机分组。结论农药毒理学试验随机分组菜单的建立,能迅速找到相关随机分组模板,完成实验动物随机分组。该方法也适用于各种非临床安全性测试的随机分组。     

运用Excel"随机数发生器"进行随机化分组

目的探讨随机化分组在Excel软件中的实现。方法运用Excel数据分析软件将受试对象按照随机原则进行分组。结果运用随机数发生器实现对实验对象进行分组。结论通过Excel随机数发生器进行随机设计分组,方法简捷灵活,具有很强的可操作性和科学性。     

计算机随机分组法在营养学实验研究中的应用

在营养学实验研究中 ,为保证实验对象的代表性和研究组间的可比性 ,经常需要将实验对象 (如动物 )随机分配入实验组和对照组。随机对照实验被认为是营养学实验方法的“金标准”[1] 。目前常用的随机化方法是将实验动物按某一特征 (如体重 )编号排序后 ,查随机数字表或随机排列表 ,按随机原则进行分组 [2 ]。这一方法涉及将体重从小到大排序 ,查随机表 ,抄录随机数 ,列分组表等过程 ,手工操作十分麻烦且容易出错 ,特别当实验对象或分组数较多时 ,更觉费时费力。虽有人使用美国 SAS软件 ,对实验对象进行计算机随机分组 [3 ] ,但由于该软件价…     

变化区组随机化及其SAS宏实现北大核心CSCD

目的非固定区组长度可有效降低随机对照临床试验(RCT)中区组随机化分组的可预测性,为此编制全自动SAS宏程序实现变化区组随机化。方法结合变化区组随机化的原理编制SAS宏程序,通过模拟实例演示程序的使用。结果输入设定的宏变量参数,运行SAS宏程序即可生成变化区组随机化的受试者分配方案。此外,该SAS宏能实现多臂不等比RCT的分配问题,提供了受试者随机分配序列重现的功能。结论多区组长度的设置和区组长度的随机选择能够降低分组可预测性,避免随机分组阶段选择偏倚的产生。本文的SAS宏为随机对照临床试验实现变化区组随机化提供了便利。     

应用电子表格Excel宏功能进行计量资料比较的显著性检验

目的探讨应用电子表格Excel宏功能进行计量资料比较的显著性检验。方法安装电子表格软件Excel，加载宏，调用数据分析工具，进行计量资料比较的方差齐性检验、随机设计的两总体均数比较的t检验、配对设计资料均数比较的t检验、方差分析等显著性检验。结果Excel进行计量资料比较的显著性检验与统计学教材一致。结论用Excel进行计量资料的显著性检验，方便直观快速，可以用于医学统计学处理。     

IL-6和IL-8在三氯乙烯致敏豚鼠血清中的变化

目的通过豚鼠致敏最大值试验(guinea pig maximization test,GPMT)方法建立豚鼠致敏模型,检测并比较TCE致敏豚鼠与未致敏豚鼠血清中IL-6和IL-8的水平,探讨IL-6和IL-8在三氯乙烯(TCE)过敏性皮炎中的作用。方法将豚鼠随机分为空白对照组,溶剂(橄榄油)对照组,DNCB阳性对照组和TCE处理组。采用GPMT法建立豚鼠致敏模型。根据致敏结果及末次激发后采血的不同时点将TCE处理组分为TCE未致敏24h组,TCE致敏24h组,TCE未致敏72h组,TCE致敏72h组。用ELISA试剂盒测定血清中IL-6和IL-8的含量。结果DNCB组致敏率为100%,TCE组致敏率为62.1%。溶剂对照组与空白对照组差异均无统计学意义。与溶剂对照组相比,TCE未致敏24h组IL-6水平降低,差异有统计学意义(P<0.05),TCE未致敏72h组与TCE未致敏24h组相比,IL-6水平明显升高,差异有统计学意义(P<0.05)。与溶剂对照组相比,TCE未致敏72组IL-8水平明显降低,差异有统计学意义(P<0.05)。结论血清中细胞因子IL-6和IL-8水平在TCE诱导的致敏组和未致敏组豚鼠间...     

三氯乙烯致敏豚鼠体内循环免疫复合物的变化

目的检测循环免疫复合物(CIC)在三氯乙烯(TCE)致敏豚鼠体内含量的变化,探讨其可能的作用。方法选用体重250～300 g的白色雌性豚鼠,随机分成空白对照组、溶剂对照组、TCE处理组。根据豚鼠最大值试验(GPMT)方法处理豚鼠。分别在实验结束后24 h、72 h采血,检测血清中IgA/C3-CIC、IgG/C3-CIC和IgM/C3-CIC的浓度。结果 TCE处理组致敏率为65.38%;与溶剂对照组相比,TCE致敏组和TCE未致敏组的血清中IgA/C3-CIC、IgG/C3-CIC和IgM/C3-CIC的浓度均明显降低,差异有统计学意义(P<0.05)。结论 TCE处理组豚鼠血清中循环免疫复合物含量降低,可能提示其体液免疫发生改变。     

临床检验质控的自动化处理

目的开展室内质控已成为现代临床实验室质量管理的手段,卫生部明确要求并规定了临床实验室质量管理的要求,实验室应当对开展的检验项目进行室内质量控制。虽然目前在我国大多数临床实验室都已随机配有室内质控软件,但所用的质控软件只能用于一种项目或一部仪器中,并不能方便地应用于所有项目或仪器。为此,笔者尝试利用Excel的强大数据和图表处理功能,结合其所提供的VBA编程语言,实现室内质控数据的自动化处理。方法将质控报表的相应项目定义在Excel工作表的不同单元格中,均值、标准差、变异系数等运算借助Excel提供的各种函数,质控图的绘制利用Excel的宏录制功能,自动绘制出质控图。结论用Excel自动处理室内质控图,图表美观灵活,运算准确而迅速,适宜在各级医院检验室应用。     

六苄基六氮杂异伍兹烷豚鼠皮肤致敏试验

目的研究六苄基六氮杂异伍兹烷（HBIW,合成单体炸药CL-20的中间体）对豚鼠的致敏性,为CL-20的危险度评价提供基础数据。方法将40只普通级豚鼠按体重随机分为3组,实验组20只,诱导期和激发期分别在两侧背部涂以一定浓度的受试物。阴性对照组10只,诱导期涂以凡士林,激发期则涂以受试物。阳性对照组10只,采用一定浓度的2,4-二硝基氯苯进行诱导和激发。实验结束后观察动物皮肤是否出现红斑或水肿,并计算致敏率。结果阳性对照组的致敏率为100%,阴性对照组为0%。实验组仅有1只豚鼠出现轻度红斑,致敏率为5%。结论 HBIW的致敏等级为Ⅰ级,具有弱致敏性。     

某种单兵武器发射对豚鼠鼓膜的损伤实验

目的验证某种单兵武器发射时冲击波对豚鼠鼓膜的损伤效应,为听觉器官冲击伤的防治提供新的实验依据。方法选取同等条件下的雄性健康豚鼠40只,随机分成实验组（30只）和对照组（10只）,均放置于单兵武器发射手的位置,其中实验组是在武器发射过程中布放,对照组在实验间隙布放,放置持续时间一致。通过对其前庭器官解剖病理学检查结果,评估其在武器发射时受冲击波影响的损伤程度。结果武器发射结束后96.7%的实验组豚鼠出现鼓膜破裂等前庭器官损伤,而对照组豚鼠没有出现损伤。结论某单兵武器发射会引起豚鼠的前庭器官损伤。     

三氯乙烯致敏豚鼠单核淋巴细胞中β-arrestin蛋白表达和核转录因子及激活蛋白-1活性的研究

目的 测定三氯乙烯(TCE)致敏豚鼠外周血单核淋巴细胞(PBMC)中β-arrestin蛋白表达水平,核转录因子(NF-κB)和激活蛋白-1(AP-1)活性以及血清中肿瘤坏死因子(TNF-α)水平,探讨TCE致敏中免疫反应调节机制.方法 根据豚鼠最大值试验方法,用TCE对豚鼠进行处理(TCE处理组,14只),同时设立空白对照(5只)和DNCB阳性对照(7只),对受试动物进行皮肤反应评分,据此判断致敏与否,并把TCE处理组分为TCE致敏组和TCE未致敏组.提取豚鼠外周血单核细胞(PBMC),用免疫蛋白印迹(Western bloting)法检测β-arrestin蛋白表达水平,用电泳迁移率试验(EMSA)方法检测NF-κB和AP-1活性.酶联免疫吸附(ELISA)试剂盒检测血清中TNF-α水平.结果 空白对照组豚鼠未见红斑或水肿,TCE组的部分豚鼠皮肤出现红斑和水肿,致敏率为71.4%,DNCB处理组所有动物皮肤出现明显红斑或水肿,致敏率为100%.空白对照、TCE未致敏组、TCE致敏组和DNCB处理组豚鼠PBMC中β-arrestin蛋白表达水平比较,差异无统计学意义(P>0.05).与空白对照组和TCE未致敏组相比,TCE致敏组NF-κB活性明显升高,且差异有统计学意义(P<0.05);而各组豚鼠AP-1活性比较,差异无统计学意义(P>0.05).TCE致敏组血清中TNF-α水平[(55.485+8.732)pg/ml]较空白对照组[(32.118±12.550)pg/ml]明显升高,差异有统计学意义(P<0.05).结论 以TCE致敏豚鼠β-arrestin和AP-1可能没被激活,而NF-κB被明显激活且在TCE致敏免疫反应中发挥着调节作用.     

Cell-mediated cytotoxicity and humoral immune response in ascorbic acid-deficient guinea pigs.

Guinea pigs were divided into three dietary groups: ascorbic-acid deficient, pair-fed, and ad libitum control. Two weeks later guinea pigs were immunized intradermally with 5 x 10(8) chicken erythrocytes in Freund's complete adjuvant. Hemagglutinating antibody titers to chicken erythrocytes 2 weeks after immunization were comparable in all three dietary groups. In vitro 51Cr release from labeled chicken erythrocyte target cells incubated with lymphoid cells from spleens of ascorbic acid-deficient guinea pigs was significantly less than with spleen cells from pair-fed and ad libitum control guinea pigs. The percentage of splenic lymphoid cells that formed rosettes with rabbit erythrocytes, a T cell marker, was the same in all three dietary groups. The defect of ascorbic acid deficiency may reflect an impairment of T lymphocytes function in cell-mediated cytotoxicity or a change in number or function of another cell type.     

高频连续噪声对豚鼠听阈及耳蜗结构的影响

目的研究高频连续噪声暴露对豚鼠听阈及耳蜗结构的影响。方法16只雄性花色豚鼠随机分为噪声暴露组和对照组,每组8只豚鼠。噪声暴露组豚鼠于噪声暴露前测定听觉脑干反应阈值(auditorybrainstem responses,ABR),每日接触中心频率为4 kHz、倍频程、100 dB(A)声压级(sound pressure level,SPL)连续噪声,每天8 h,连续7 d;末次噪声暴露结束后第8天,测定ABR后处死豚鼠,取耳蜗,进行组织病理学、透射电镜及扫描电镜的观察。对照组豚鼠不接触噪声,于相应时间点测定ABR,其他处理同噪声暴露组。结果噪声暴露组豚鼠在2,4,8 kHz处的阈移分别为13.4,31.3,31.9,耳蜗外毛细胞静纤毛发生散在缺失、倒伏,胞浆萎缩、空化,细胞核萎缩或消失,部分线粒体消失,个别线粒体发生髓样变。结论噪声暴露可提高豚鼠的听阈并对其耳蜗结构具有损伤作用。     

三氯乙烯染毒对豚鼠肝功能与凋亡基因表达的影响

目的探讨使用三氯乙烯(TCE)染毒对豚鼠肝功能和肝细胞凋亡基因(BAX、BAD、Bc1-2)表达的影响。方法将24只豚鼠随机分为3组,采用豚鼠最大值法(GPMT),设立TCE实验组、阴性对照组、阳性对照组,用皮内注射的方式分别注射TCE、橄榄油、2,4-二硝基氯苯(DNCB),实验结束后观察动物皮肤改变,应用自动生化分析仪检测动物肝功能指标,用荧光定量PCR检测肝细胞凋亡基因表达水平。结果 TCE实验组和阳性对照组动物出现明显皮肤损害。TCE实验组动物血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、乳酸脱氢酶(LDH)活力明显高于阴性对照组(P0.05或P0.01)。肝细胞BAX、BAD的mRNA表达水平比阴性对照组显著升高,Bc1-2表达水平下降(P0.05或P0.01)。结论三氯乙烯可诱导豚鼠产生明显的皮肤变态反应,引起实验动物肝功能指标改变和肝细胞凋亡基因表达水平明显改变。     

Dietary phosphate deprivation increases renal synthesis and decreases renal catabolism of 1,25-dihydroxycholecalciferol in guinea pigs.

In guinea pigs, dietary phosphate deprivation decreases plasma phosphate concentration, increases plasma 1.25-dihydroxycholecalciferol [1,25-(OH)2D3] concentration and causes hypercalcemia concurrent with the maximal increase in plasma 1,25-(OH)2D3 levels. Our objective was to determine whether increased synthesis or decreased catabolism contributed to the elevation in plasma 1,25-(OH)2D3. Preliminary experiments using renal mitochondria from guinea pigs fed a control diet revealed that 23,25-dihydroxycholecalciferol [23,25-(OH)2D3], not 24,25-dihydroxycholecalciferol [24,25-(OH)2D3], was the reciprocal side-chain metabolite to 1,25-(OH)2D3 in this species. An assay employing guinea pig renal mitochondria was used to measure the renal synthesis of 1,25-(OH)2D3 and 23,25-(OH)2D3 from [3H]25-OH-D3. These metabolites were unequivocally identified by combinations of HPLC, ultraviolet spectrophotometry and mass spectrometry. This renal mitochondrial assay was subsequently used to investigate the effect of dietary phosphate deprivation on guinea pig vitamin D metabolism. Within 1 wk the rate of synthesis of 1,25-(OH)2D3 was maximal in phosphate-deprived guinea pigs. This rate was significantly (P less than 0.005) higher than that achieved in same-day control guinea pigs. Conversely, within 1 d the synthesis of 23,25-(OH)2D3 was significantly (P less than 0.005) decreased in phosphate-deprived guinea pigs. Similarly, the rate of 1,25-(OH)2D3 metabolism was decreased within 1 d of dietary phosphate deprivation and was at a minimum within 1 wk. This rate was significantly (P less than 0.005) less than that attained in same-day control guinea pigs. These results suggest that both increased synthesis and decreased metabolism of 1,25-(OH)2D3 contribute to the plasma 1,25-(OH)2D3 elevation that occurs in response to dietary phosphate deprivation.     

Genetic polymorphism in Taenia solium cysticerci recovered from experimental infections in pigs.

Taenia solium cysticerci recovered from naturally infected pigs from Mexico, Honduras and Tanzania show a clonal structure and local lineages with probable events of genetic recombination without genetic flow within them, as revealed by RAPD. To evaluate genetic polymorphism from cysticerci recovered from experimental infections, 4 pigs were infected with T. solium eggs obtained from tapeworms released by 3 human carriers, a 10-year-old female, a 25-year-old female, and a 44-year-old male, the 4th pig was infected with a mixture of eggs from the 3 tapeworms. Each pig was orally inoculated with 50,000 eggs. After 16 weeks pigs were humanely euthanized and cysticerci were excised. Parasites recovered from each pig were analyzed by RAPD. The proportion of polymorphic loci and the mean heterozygosity as well as a dendogram and an analysis of principal coordinate and minimum spanning tree were obtained. All four pigs developed viable cysticerci; the percent infection was obtained from the ratio of the number of eggs used for infection and the number of cysticerci counted in each pig after necropsy. Infection varied from 0.2 to 4.2%. The values obtained for the proportion of polymorphic loci (0.14-0.55) and the average of expected heterozygosity (0.06-0.22) in the present experimental infection had a broader range than those reported in the literature from natural infections. The dendogram obtained clustered cysticerci into two main groups; the minimum spanning tree allowed to corroborate the data obtained in the dendogram and gave a better discrimination because in a three-dimensional plot it was easier to see that all cysticerci from each tapeworm were clustered amongst themselves. The results obtained could be hypothetically explained because environmental factors and genetic selection agents present in nature influence natural infections but do not participate in experimental ones.     

Immunity induced by DNA immunization with herpes simplex virus type 2 glycoproteins B and C

The complete sequence of herpes simplex virus type 2 (HSV-2) glycoproteins B and C (gB & gC) were cloned into plasmid expression vectors and evaluated in murine and guinea pig genital HSV-2 models. Balb/c mice were immunized with either pgB-2 or pgC-2 plasmids intramuscularly (IM) or intradermally (ID). The vaccines induced HSV-2-specific neutralizing and ELISA IgG antibody, but little or no enhancement of viral clearance from the vagina was detected following intravaginal challenge. Immunization of guinea pigs with pgB-2 or pgC-2 induced ELISA IgG antibody; however, antibody titers were approximately one log(10) unit lower than that seen in HSV-2 convalescent sera. IM immunization of guinea pigs with either plasmid also did not decrease vaginal viral shedding following vaginal challenge, but the severity of the acute disease and the subsequent number of recurrent lesion days were reduced in animals immunized with pgB-2. Lastly, IM immunization of latently infected guinea pigs with a combined gB-2 and gC-2 plasmid vaccine significantly reduced the number of subsequent HSV-2 recurrences. DNA vectors expressing gB-2 or gC-2 were both immunogenic, although the gB-2 plasmid induced higher titers of antibody and significantly reduced primary and recurrent herpetic disease in the guinea pig model. These results also suggest that immunotherapy with plasmid expression vectors may be effective against recurrent genital HSV-2 disease.