彗星试验检测酚类化合物对小鼠脾脏细胞DNA损伤的研究

应用彗星试验技术 ,研究了酚类化合物对小鼠脾脏细胞DNA的损伤作用 ,试验结果表明 ,12种酚类化合物对小鼠脾脏细胞均能引起不同程度的DNA损伤 ,并呈现剂量效应关系。与对照组相比 ,均有显著性差异 (P <0 0 5 ,P <0 0 1)。试验结果亦表明 。

氨基酚类化合物对人淋巴细胞及小鼠脾脏细胞DNA损伤的比较

目的研究氨基酚类化合物对人外周血淋巴细胞及小鼠脾脏细胞DNA的损伤效应。方法选用人淋巴细胞及小鼠脾脏细胞进行研究。暴露时间为1h。应用单细胞凝胶电泳技术,计算细胞损伤率及专用单位。结果3种氨基酚类化合物均能引起两种细胞不同程度的DNA损伤,高剂量组与阴性对照组相比,差异均有显著性(P<0.05),其中对氨基酚的毒性高于邻氨基酚和间氨基酚,其损伤的程度随剂量的增加而增加。结论与小鼠脾脏细胞相比,人外周血淋巴细胞的敏感性较强,更能直接反映氨基酚类化合物对人群的遗传毒性。

穿龙薯蓣皂苷提取及其对小鼠脾脏细胞活性的影响

利用回流提取法分别提取穿龙薯蓣粉末和穿龙薯蓣粗块,并利用石油醚、乙酸乙酯、水饱和正丁醇三种有机溶剂对提取液进行萃取,再利用这四种提取液对小鼠脾脏细胞活力进行检测试验.结果表明:薯蓣皂苷在水中的溶解性较其他三种有机溶剂好,穿龙薯蓣表面积越大,越有利于薯蓣皂苷的提取.MTT试验表明,穿龙薯蓣皂苷对小鼠脾脏细胞活力的影响不明显.

两种胃癌细胞株与小鼠原代脾脏淋巴细胞共培养对Th17细胞分化的影响

目的观察不同胃癌细胞株与CD4^(+)T细胞共培养后Th17细胞分化的改变情况。方法将分离并鉴定后的小鼠原代脾脏淋巴细胞分别与人胃癌SGC-7901细胞株、人胃癌BGC-823细胞株共培养,分为SGC-7901共培养组、BGC-823共培养组和对照组。以细胞内细胞因子染色FCM测定Th17细胞,计算CD4^(+)IL-17A^(+)细胞数量百分比。结果SGC-7901共培养组、BGC-823共培养组、对照组的CD4^(+)IL-17A^(+)细胞数量分别为(3.97±0.27)%、(5.17±0.14)%、(1.96±0.29)%,与SGC-7901共培养组比较,BGC-823共培养组Th17细胞数量均显著增多,差异有统计学意义(P<0.05)。结论在胃癌环境下,CD4^(+)T细胞向Th17细胞分化增多。

蒙药特润舒都乐对模型小鼠脾脏NK细胞活性的影响

目的：观察和探讨蒙药特润舒都乐对小鼠脾脏NK细胞杀伤活性的影响，了解该蒙药的生物学活性。方法：采用皮下给药的体内实验及3H-TdR掺入抑制法检测小鼠脾脏NK细胞活性。结果：蒙药特润舒都乐能显着提高小鼠脾脏NK细胞杀伤活性，且呈剂量依赖性，其高、中、低剂量（50，25，10mg／L）各组的NK活性分别为64．39±16．53、46．21±13．28、36．09±12．26，与空白对照及阴性对照均有显着性差异（P〈0．05）。结论：蒙药特润舒都乐能提高NK细月缸活性，并有剂量依赖性。

羧胺三唑增强小鼠脾脏淋巴细胞促进结肠癌细胞系MC38和C26凋亡

目的探讨羧胺三唑(CAI)直接或通过小鼠脾脏淋巴细胞间接对小鼠结肠癌细胞系(MC38和C26)存活和凋亡的影响。方法体外培养结肠癌细胞系MC38和C26细胞,分离小鼠脾脏淋巴细胞,MC38和C26细胞分别与小鼠脾脏淋巴细胞共培养,不同浓度CAI处理细胞。CCK-8法检测细胞存活,流式细胞测量术检测细胞凋亡以及小鼠脾脏淋巴细胞中CD4^(+)CD3^(+)T细胞和CD8^(+)CD3^(+)T细胞的比例。结果不同浓度CAI处理MC38和C26细胞后,活细胞数目减少(P<0.05、P<0.01和P<0.001),凋亡率升高(P<0.01和P<0.001);MC38和C26细胞分别与小鼠脾脏淋巴细胞共培养48 h后,MC38和C26细胞数目减少(P<0.001),凋亡率升高(P<0.001);不同浓度CAI处理小鼠脾脏淋巴细胞共培养条件下的MC38和C26细胞48 h,与单独小鼠脾脏淋巴细胞处理相比,MC38和C26细胞数目进一步减少(P<0.001),凋亡率进一步升高(P<0.05和P<0.001);不同浓度CAI处理小鼠脾脏淋巴细胞48 h后,小鼠脾脏淋巴细胞中CD4^(+)CD3^(+)T细胞和CD8^(+)CD3^(+)T细胞的比例显著升高(P<0.05和P<0.01)。结论CAI抑制MC38和C26细胞存活、促进细胞凋亡,并可能通过升高小鼠脾脏淋巴细胞中CD4^(+)CD3^(+)T细胞和CD8^(+)CD3^(+)T细胞的比例增强小鼠脾脏淋巴细胞抑制MC38和C26细胞存活、促进细胞凋亡。

树突状细胞肿瘤疫苗研究进展

树突状细胞的发现和特性 1973年，Ralph Steinman和Zanvil Cohn等从小鼠脾脏细胞中分离出一群外形奇特的细胞，这些细胞在成熟时伸出许多树突样或伪足样突起，因此命名为树突状细胞（dendritic cell，DC）。研究发现，树突状细胞具有强大的活化初始T细胞的能力，是适应性免疫应答的主要启动者。

全淋巴结照射对小鼠半相合造血干细胞移植影响的实验研究

目的探讨全淋巴结照射(TLI)作为移植预处理方案的可行性。方法 (1)将1.8周龄健康清洁级雌性 C57BL/6(H-2b)小鼠与8周龄健康清洁级雄性 BALB/C(H-2d)小鼠按2:1同居法杂交,培育(C57BL/6×BALB/C)F1(H-2b/d)子一代小鼠。(2)(C57BL/6×BALB/C)F1(H-2b/d)小鼠分2组,分别接受全淋巴结照射(TLI。

Diminution of toxic copper accumulation in toxic milk mice modeling Wilson disease by embryonic hepatocyte intrasplenic transplantation

AIM: To observe the therapeutic effect of intrasplenic transplantation with embryonic hepatocytes on amelioration of hereditary copper accumulation in toxic milk (TX) mouse modeling Wilson disease. METHODS: Donor hepatocytes were harvested from 14-d fetal liver of a pregnant homogeneous DL mouse. These cells were successively cultured, labeled with fluorescein dye Hoechst 33342 for 24 h, and sequentially infused into the spleen parenchyma of the recipient TX mice. No host immunosuppression measures were taken. Two and four weeks after transplantation, the recipients were killed for routine histologic investigation and immunohistochemistry study up to 4 wk after transplantation. The serum copper and ceruloplasmin concentrations of the recipient mice were determined by graphite furnace atomic absorption spectroscopy.RESULTS: In the following 2nd and 4th wk after transplantation, the donor hepatocytes could be visualized in the livers of 47.3% recipients. The serum ceruloplasmin and copper concentrations increased by 1.6-fold after 2 wk and 2.0-fold times after 4 wk respectively, which ultimately rose from about 30% of the normal level to nearly 60%(P＜0.01). The hepatic copper concentration decreased 7.2%, 4 wk after transplantation. Pathologic examination showed that there were many actively proliferative hepatocyte precursor cells with specific embryonic hepatocyte marker AFP migrated into hepatic sinusoidsof the recipients. A large number of cells carrying hepatocytes marker and albumin were observed in the recipient spleen tissues.CONCLUSION: Embryonic hepatocytes are capable of differentiating into mature hepatocytes in vivo. After transplantation, the hereditary abnormalities of copper metabolism in TX mice could be corrected partially by intrasplenic transplantation of homogeneous embryonic hepatocytes.

Effect of operation-synchronizing transfusion of apoptotic spleen cells from donor rats on acute rejection of recipient rats after liver transplantation

AIM: To study effect of operation-synchronizing transfusion of apoptotic spleen cells from donor rats on acute rejection of recipient rats after liver transplantation. METHODS: Two of Wistar rats were chosen randomly for normal liver pathology control and ten of SD rats chosen randomly for liver function control as blank group (no operation). The rest of Wistar and SD rats were divided into four groups: control group (only liver transplantation), Dex group (donors receiving intraperitoneal injection of dexamethasone), SpC group (recipients receiving infusion of spleen cells of donors), Dex-SpC group (recipients receiving infusion of apoptotic spleen cells of donors), with each group except blank group, containing 10 SD rats and 10 Wistar rats, respectively. Wistar rats received liver transplantation from SD rats, in the meantime they received infusion of spleen cells of donors, which were induced by an intraperitoneal injection of dexamethasone (3 mg/(d.kg)·b.w) for three days before liver transplantation. The serum alanine transaminase (ALT), total bilirubin (T bili), liver pathological changes and survival time were analysed. Statistical analysis was carried out using SPSS 10.0 for Windows. Differences of the parametric data of ALT in means were examined by one-way ANOVA. Differences of ALT between two groups were examined by LSD. Differences of the nonparametric data of T bili in means and scores of pathology classification for acute rejection were examined by Kruskal-Willis H test. The correlations between ALT and T bili were analysed by Bivariate. Kaplan-Meier curves were used to demonstrate survival distribution. The log-rank test was used to compare the survival data. RESULTS: There were significant differences in ALT of the five groups (F= 23.164 P= 0.000), and ALT in Dex-SpC group was significantly higher than that in blank control, control, Dex, and SpC groups (P = 0.000), and ALT in SpC group was significantly higher than that in blank control (P= 0.000), control (P= 0.004), and Dex groups (P= 0.02). Results of nonparametric analysis of T bill showed that there were differences in T bill of the five groups (X2= 33.265 P= 0.000). T bili in Dex-SpC group was significantly higher than that in blank control, control, Dex, and SpC groups. T bili in SpC group was higher than that in blank control, control, and Dex groups. There were significant differences in scores of pathology classification for acute rejection in each of the groups (X2= 25.933, P= 0.000). The pathologically more serious acute rejection was found in Dex-SPC group than in other groups. No sign of acute rejection was observed in the blank control group. Slight acute rejection was observed in the control group. Slight-moderate acute rejection was observed in the Dex group. Moderate-acute rejection was observed in the SpC group. Severe-acute rejection was observed in the Dex-SpC group. The survival time in Dex-SpC group was shorter than in other groups (statistic = 11.13, P= 0.011). ALT and T bili were positively correlated (r= 0.747, P= 0.000, two-tailed). CONCLUSION: In order to reduce quantity of blood loss from rats after liver transplantation, only one of ALT or T bili is needed for liver function measurement of rats. Simultaneous injection of apoptotic spleen cells from donors induced by dexamethasone to liver transplantation rats aggravates acute rejection. One important mechanism of aggravation of acute rejection may be that apoptotic cells are not removed in time and that dead cells including apoptotic cells release inflammatory factors.

The effect of spleen on the change of Treg cells during mouse pregnancy

Objective:To analyze the effect of spleen on the Treg cells during pregnancy. Methods: The mononuclear cells were separated from the peripheral, spleen and uterus or placenta blood. Flow cytometry was employed to analyze the percent of Treg cells in total T cells in different stages of pregnancy. Immunohistochemical staining was used to make sure the distribution of Treg in spleen in different stages of pregnancy. Results: The results of immunohistochemical staining showed that compared with spleen Treg cells in normal unpregnant mice, spleen Treg cells on day 7 and 14 of pregnancy significantly increased. After splenectomy, peripheral blood and placenta Treg cells on day 7 of pregnancy markedly decreased as compared with the normal pregnancy(P<0.01). And the cells on day 14of pregnancy were markedly recovered as compared with the normal pregnancy. Conclusion: Our study indicated that the spleen and its Treg cells might play important roles in transient tolerance during pregnancy.

人外周血淋巴细胞增殖试验的优化及其应用

目的:优化人外周血淋巴细胞增殖试验条件;考察动物源材料的Gal抗原含量与淋巴细胞增殖效应的相关性;并比较人外周血淋巴细胞与小鼠脾脏淋巴细胞增殖试验对动物源材料的敏感性差异。方法:从细胞接种量,阳性对照(刀豆球蛋白A,Con A)的工作浓度与细胞培养时间,优化人外周血淋巴细胞增殖试验的最佳条件;利用优化的试验方法,通过人外周血淋巴细胞与动物源性材料的(牛跟腱或由牛跟腱纯化的胶原蛋白海绵)匀浆液或浸提液共培养,考察材料的不同前处理方式对人外周血淋巴细胞增殖的影响;参考标准YY/T 1561-2017,检测材料匀浆液或浸提液的Gal抗原含量,分析其抗原含量与淋巴细胞增殖效应的相关性;同时参考标准YY/T 1465.1-2016,考察材料匀浆液与浸提液对小鼠脾脏淋巴细胞增殖效应的影响,并对比分析对动物源材料的敏感性差异。结果:人外周血淋巴细胞增殖试验最佳反应条件经优化确定为:细胞接种量为1×10^5,阳性对照(Con A)的终浓度选择5~10μg·mL^-1,培养时间为4 d。人外周血淋巴细胞与胶原蛋白海绵(匀浆组与浸提液组)共培养后,样品组与正常细胞对照组相比不存在明显差异;与牛跟腱匀浆液共培养后,与正常细胞对照组相比不存在显著性差异。胶原蛋白海绵(匀浆组与浸提液组)对小鼠脾脏淋巴细胞均无明显增殖效应。牛跟腱匀浆液在5倍稀释与50倍稀释后,对小鼠脾脏淋巴细胞均有明显增殖效应(增殖率为143.20%和128.71%)。牛跟腱匀浆液的Gal抗原含量为(3.98±1.06)×10^13个·mg^-1、胶原蛋白海绵匀浆液为(2.24±0.60)×10^13个·mg^-1、胶原蛋白海绵浸提液为(1.89±0.64)×10^13个·mg^-1。结论:淋巴细胞增殖效应与动物源性生物材料中残留Gal抗原含量的正向相关性不强。与小鼠脾脏淋巴细胞增殖试验相比,采用人外周血淋巴细胞增殖试验评价含Gal抗原的动物源性生物材料细胞免疫的敏感性未见优势。淋巴细胞增殖试验是评价动物源生物材料细胞免疫的可用方法之一。