SIAF对病毒性心肌炎模型鼠的治疗作用

目的 研究梅花鹿免疫细胞活性因子(SIAF)对病毒性心肌炎的治疗作用。方法 采用柯萨奇B3型病毒(CB_3 V)性心肌炎小鼠模型,检测心肌酶、心电图,观察心肌病理改变及死亡率。采用MTT法测定淋转及NK细胞毒活性。结果 SIAF能明显改善心肌损伤模型鼠的心电图及心肌病理损害;降低心肌酶含量;调节淋转及NK细胞毒活性。结论 SIAF对病毒性心肌炎具有一定治疗作用。     

梅花鹿免疫细胞活性因子对乙醇型胃粘膜损伤模型鼠的保护作用

目的　观察梅花鹿免疫细胞活性因子 (SIAF)对小鼠乙醇型胃粘膜损伤的保护作用。方法　制备小鼠乙醇型胃粘膜损伤模型 ,测定溃疡指数 ,观察胃的出血情况 ,免疫组化染色检测胃粘膜EGFR的表达 ,用酶法测定血清中NO的含量。结果　SIAF组的溃疡指数、胃出血情况与模型组相比 ,差异均有显著意义。SIAF组小鼠损伤周围胃粘膜EGFR表达与模型组相比明显增强。该组小鼠血清NO含量与模型组相比也明显增高。结论　SIAF具有显著的保护此种胃粘膜损伤的作用     

垂盆草提取液对小鼠免疫和抗氧化功能的研究

目的研究垂盆草提取液对实验性急性肝损伤小鼠免疫和抗氧化功能的影响。方法通过注射环磷酰胺(CP)抑制小鼠的免疫细胞,达到降低淋巴细胞增殖水平和网状内皮系统的吞噬功能,观察药物对小鼠免疫功能的调节作用;给小鼠注射D-半乳糖(D-gal),造成实验性急性肝损伤模型,测定血清及肝脏中超氧化物歧化酶(SOD)活力和丙二醛(MDA)浓度,观察垂盆草提取液对小鼠抗氧化能力的影响。结果垂盆草提取液能明显促进免疫抑制状态小鼠的淋巴细胞增殖及网状内皮系统的吞噬功能,垂盆草提取液对D-gal所致小鼠急性肝损伤模型具有提高SOD活性和降低MDA浓度的作用。结论垂盆草提取液对免疫功能低下的小鼠具有增强作用,同时能提高实验性肝损伤小鼠的抗氧化能力。     

皖南红豆杉枝叶中总黄酮抗肿瘤活性研究

以MTT法考察皖南红豆杉枝叶中总黄酮的体外抗肿瘤活性;以小鼠移植瘤模型测定总黄酮对荷瘤小鼠抑瘤率和腹腔巨噬细胞吞噬能力,研究其体内抗肿瘤活性和免疫调节作用。结果表明,总黄酮对S180细胞生长有显著抑制作用,呈现剂量依赖关系。体内试验表明,与生理盐水组相比,总黄酮对小鼠S180实体瘤有显著抑制作用（P〈0.01）,可提高荷瘤小鼠的脾指数。与环磷酰胺组相比,总黄酮可使荷S180实体瘤小鼠腹腔巨噬细胞吞噬能力显著升高（P〈0.01）,提示其对小鼠无免疫抑制作用。     

蒲公英黄酮对小鼠DSS诱导性肠炎的功能恢复研究

选取蒲公英活性成分黄酮作为抗炎和调节肠道微生态平衡的研究对象,建立小鼠结肠炎模型,通过分析蒲公英活性成分对结肠炎小鼠体重变化、血常规生化指标、炎症因子、组织病理学、等指标的影响,阐明蒲公英活性成分黄酮对小鼠UC的治疗作用。     

三肽D-Trp-Arg-Leu-NH2抑制B16黑色素瘤细胞黑色素合成的机制

为了研究三肽D-Trp-Arg-Leu-NH₂(dWRL)对小鼠B16细胞的酪氨酸酶(TYR)活性、黑色素合成及相关基因和蛋白表达的影响，探讨dWRL抑制黑色素生成的可能机制，我们体外培养小鼠B16细胞，采用MTT法测定细胞增殖情况、L-多巴氧化法测定TYR活性、氢氧化钠裂解法测定细胞黑色素含量、qRT-PCR和Western Blot法分别检测药物作用后B16细胞中小眼畸形相关转录因子(MITF)、TYR的mRNA和蛋白表达水平。结果显示三肽dWRL在试验浓度范围内可明显抑制α-黑色素细胞刺激素(α-MSH)诱导的B16细胞内TYR活性、黑色素合成量及MITF、TYR基因及蛋白的表达量，且呈浓度依赖性。所以三肽dWRL在体外能抑制B16细胞黑色素的合成，上述变化可能是通过下调MITF和TYR的基因转录和蛋白表达作用来完成。     

RGD特异性结合多肽与重组改构人肿瘤坏死因子融合基因的克隆表达及其抑瘤活性

目的构建RGD多肽与重组改构人肿瘤坏死因子融合基因,进一步提高rmhTNF的临床疗效。方法应用基因重组技术构建了CRGDC与rmhTNF的融合基因,在E.coliDH5α中诱导表达重组蛋白RGDrmhTNF,采用溶菌酶法裂解菌体,裂解上清经硫酸铵沉淀、阴阳离子交换层析纯化后,用S180荷瘤小鼠模型和L929细胞体外杀伤实验测定纯化的重组蛋白RGDrmhTNF的体内、外杀伤活性。结果正确构建了编码RGDrmhTNF融合基因,重组工程菌升温诱导后以部分可溶形式表达RGDrmhTNF,纯化后纯度为96.88%,体外对L929细胞杀伤作用的比活性为3.6×108IUmg,与rmhTNF相当(3.0×108IUmg)。在S180荷瘤小鼠模型中,当RGDrmhTNF剂量为12×105IUkg时,对肿瘤的生长抑制率为84.9%,显著高于相同剂量的rmhTNF(肿瘤抑制率为59.09%,P<0.01)和环磷酰胺组(肿瘤抑制率为71.21%,P<0.01)。结论重组蛋白RGDrmhTNF可以提高rmhTNF的治疗效果。     

双靶点抗组胺化合物的设计、合成及活性研究

以H1受体拮抗剂地氯雷他定为基本骨架,采用药效团和生物电子等排原理,设计合成了3个新型具有H1/H4双重拮抗活性的化合物,并对其进行了体外H1/H4靶点活性检测、脂多糖(LPS)致小鼠急性炎症因子释放和组胺诱导小鼠皮肤血管通透性实验研究。实验结果显示,3个化合物均具有H1/H4双重拮抗活性和显著的抗炎、抗过敏活性,且可减少LPS诱导小鼠TNF-α释放(P0. 01),抑制组胺引起的毛细血管通透性增高(P0. 01),抗炎、抗过敏活性优于地氯雷他定和卢帕他定,具有很高的开发价值。     

梅花鹿鹿茸活性多肽的提取及免疫功效的初步研究

目的提取并分离纯化不同加工方法处理的梅花鹿鹿茸活性多肽(PAP),并对其免疫功效进行初步研究。方法以不同工艺处理的鹿茸为原料,应用分子排阻层析和离子交换层析提取鹿茸活性多肽,通过淋巴细胞增殖、细胞因子产生等试验,检测PAP对淋巴细胞的免疫功效。结果所提取的梅花鹿鹿茸活性多肽蛋白含量分别为:冻干茸9.20 mg/ml;冷冻鲜茸1.30 mg/ml;热炸茸1.10 mg/ml。鹿茸多肽可以促进小鼠T、B淋巴细胞的增殖,活化巨噬细胞分泌IL-12,对机体有免疫增强作用。结论梅花鹿鹿茸活性多肽对机体的细胞免疫有显著增强作用。     

小鼠骨髓间充质干细胞对异基因肝移植大鼠淋巴细胞增殖的抑制作用

目的观察小鼠骨髓间充质干细胞(Mesenchymal stem cells,MSCs)移植后,在异基因肝移植模型大鼠胸腺、脾脏及骨髓中的迁徙、定居情况及对淋巴细胞增殖的抑制作用,以探讨MSCs诱导异基因移植免疫耐受的可能性。方法体外分离、纯化并培养小鼠MSCs;将小鼠肝组织块埋入大鼠肝脏切口,建立异基因肝移植大鼠模型;经尾静脉移植DAPI标记的小鼠MSCs,通过激光共聚焦显微镜观察24 h及5 d时,MSCs在模型大鼠胸腺、脾脏及骨髓内的迁徙及定居;采用体外淋巴细胞增殖实验检测小鼠MSCs对异基因肝移植大鼠淋巴细胞增殖的抑制作用。结果MSCs移植后,24 h即散在分布于胸腺、脾脏及骨髓内,5 d时集中在血管周围;经MSCs治疗后,大鼠胸腺和脾脏淋巴细胞增殖均受到抑制,与未经治疗的模型大鼠相比,差异有显著意义。结论小鼠MSCs可迁徙至异基因肝移植大鼠免疫器官内,并具有抑制淋巴细胞增殖的作用。     

Identification of Stereochemical Configurations of Cyclopenta[ cd ]pyrene DNA Adducts in Strain A/J Mouse Lung and C3H10T1/2CL8 Cells

Four major and several minor DNA adducts were resolved by 32 P-postlabeling analysis of DNA from strain A/J mouse lung and C3H10T1/2CL8 (C3H10T1/2) mouse embryo fibroblasts treated with cyclopenta[ cd ]pyrene (CPP). The identical pattern of adducts was seen in vivo and in vitro. Cochromatography of synthetic resolved diastereomers of cis - and trans - N 2 -CPP-deoxyguanosine-3'-phosphates with the in vivo adducts obtained from strain A/J mouse lung and in vitro adducts obtained from C3H10T1/2 cells allowed identification of the predominant DNA adduct as cis -(3 R ,4 S )- N 2 -CPP-deoxyguanosine. The second most abundant adduct formed in vivo and in vitro was identified as trans -(3 S ,4 S )- N 2 -CPP-deoxyguanosine.     

Postnatal Changes of Neural Stem Cells in the Mammalian Auditory Cortex

Our previous study reported neural stem cells (NSCs) in the auditory cortex (AC) of postnatal day 3 (P3) mice in vitro. It is unclear whether AC-NSCs exist in vivo. This study aims to determine the presence and changes of AC-NSCs during postnatal development and maturation both in vitro and in vivo. P3, postnatal day 14 (P14), 2-month-old (2M), and 4-month-old (4M) mouse brain tissues were fixed and cryosectioned for NSC marker immunostaining. In vitro, P3, P14, and 2M AC tissues were dissected and cultured in suspension to study NSCs. NSC proliferation was examined by EdU incorporation and cell doubling time assays in vitro. The results show that Nestin and Sox2 double expressing NSCs were observed in the AC area from P3 to 4M in vivo, in which the number of NSCs remarkably reduced with age. In vitro, the neurosphere forming capability, cell proliferation, and percentage of Nestin and Sox2 double expressing NSCs significantly diminished with age. These results suggest that AC-NSCs exist in the mouse AC area both in vitro and in vivo, and the percentage of AC-NSCs decreases during postnatal development and maturation. The results may provide important cues for the future research of the central auditory system.     

Injurious Effects of Curcumin on Maturation of Mouse Oocytes, Fertilization and Fetal Development via Apoptosis

Curcumin, a common dietary pigment and spice, is a hydrophobic polyphenol derived from the rhizome of the herb Curcuma longa. Previously, we reported a cytotoxic effect of curcumin on mouse embryonic stem cells and blastocysts and its association with defects in subsequent development. In the present study, we further investigated the effects of curcumin on oocyte maturation and subsequent pre- and post-implantation development, both in vitro and in vivo. Notably, curcumin induced a significant reduction in the rate of oocyte maturation, fertilization, and in vitro embryonic development. Treatment of oocytes with curcumin during in vitro maturation (IVM) led to increased resorption of postimplantation embryos and decreased fetal weight. Experiments with an in vivo mouse model disclosed that consumption of drinking water containing 40 μM curcumin led to decreased oocyte maturation and in vitro fertilization as well as early embryonic developmental injury. Finally, pretreatment with a caspase-3-specific inhibitor effectively prevented curcumin-triggered injury effects, suggesting that embryo impairment by curcumin occurs mainly via a caspase-dependent apoptotic process.     

Dihydrolipoic Acid Induces Cytotoxicity in Mouse Blastocysts through Apoptosis Processes

α-Lipoic acid (LA) is a thiol with antioxidant properties that protects against oxidative stress-induced apoptosis. LA is absorbed from the diet, taken up by cells and tissues, and subsequently reduced to dihydrolipoic acid (DHLA). In view of the recent application of DHLA as a hydrophilic nanomaterial preparation, determination of its biosafety profile is essential. In the current study, we examined the cytotoxic effects of DHLA on mouse embryos at the blastocyst stage, subsequent embryonic attachment and outgrowth in vitro, in vivo implantation by embryo transfer, and early embryonic development in an animal model. Blastocysts treated with 50 μM DHLA exhibited significantly increased apoptosis and a corresponding decrease in total cell number. Notably, the implantation success rates of blastocysts pretreated with DHLA were lower than that of their control counterparts. Moreover, in vitro treatment with 50 μM DHLA was associated with increased resorption of post-implantation embryos and decreased fetal weight. Data obtained using an in vivo mouse model further disclosed that consumption of drinking water containing 100 μM DHLA led to decreased early embryo development, specifically, inhibition of development to the blastocyst stage. However, it appears that concentrations of DHLA lower than 50 μM do not exert a hazardous effect on embryonic development. Our results collectively indicate that in vitro and in vivo exposure to concentrations of DHLA higher than 50 μM DHLA induces apoptosis and retards early pre- and post-implantation development, and support the potential of DHLA to induce embryonic cytotoxicity.     

Hazardous Effects of Curcumin on Mouse Embryonic Development through a Mitochondria-Dependent Apoptotic Signaling Pathway

In this study, we examined the cytotoxic effects of curcumin, the yellow pigment of Curcuma longa, on the blastocyst stage of mouse embryos, subsequent embryonic attachment, and outgrowth in vitro and in vivo implantation by embryo transfer. Mouse blastocysts were incubated in medium with or without curcumin (6, 12 or 24 μM) for 24 h. Cell proliferation and growth were investigated using dual differential staining, apoptosis was analyzed with terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), and implantation and post-implantation development of embryos were measured by in vitro development analysis and in vivo embryo transfer, respectively. Blastocysts treated with 24 μM curcumin displayed significantly increased apoptosis and decreased total cell number. Interestingly, we observed no marked differences in the implantation success rates between curcumin-pretreated and control blastocysts during in vitro embryonic development through implantation with a fibronectin-coated culture dish. However, in vitro treatment with 24 μM curcumin was associated with decreased implantation rate and increased resorption of postimplantation embryos in mouse uterus, as well as decreased fetal weight in the embryo transfer assay. Our results collectively indicate that in vitro exposure to curcumin triggers apoptosis and retards early postimplantation development after transfer to host mice. In addition, curcumin induces apoptotic injury effects on mouse blastocysts through ROS generation, and further promotes mitochondria-dependent apoptotic signaling processes to impair sequent embryonic development.     

卡介苗溶菌黏膜免疫调理剂对实验性免疫功能低下小鼠的免疫调节作用

目的观察卡介苗溶菌黏膜免疫调理剂(BCG-b)对实验性免疫功能低下小鼠的免疫调节作用。方法将BALB/c小鼠分为BCG-b高、中、低剂量组,每只分别隔日灌服BCG-b20、5和1U/0.2ml;阳性对照组每只隔日灌服阿胶液0.2ml,阴性对照组每只隔日灌服生理盐水0.2ml;正常对照组不给任何药物。除正常对照组外,其余各组均服药10次,并于首次服药后第14日皮下注射环磷酰胺,100mg/kg,建立免疫功能低下小鼠模型。以碳粒廓清法、血清溶血素测定法及3H-TdR掺入法分别检测各组小鼠单核-巨噬细胞吞噬功能、特异性体液免疫功能以及T淋巴细胞转化功能。结果与阴性对照组相比,BCG-b3个不同剂量组小鼠单核-巨噬细胞的吞噬功能和特异性抗体血清溶血素含量明显提高,小鼠T淋巴细胞转化功能呈双向性调节作用,即高、中剂量表现为抑制作用,低剂量表现为促进作用。结论BCG-b对实验性免疫功能低下小鼠具有明显的免疫调节作用。     

The Involvement of the Endogenous Opioid System in the Gastrointestinal Aging in Mice and Humans

Nearly 20% of elderly patients suffer from constipation, but the age-related changes in the gastrointestinal (GI) tract remain insufficiently elucidated. In this study, the alterations within the endogenous opioid system (EOS) as a potential cause of constipation in the elderly were evaluated. The GI functions were assessed in vitro and in vivo and compared between 6-, 12- and 18-month old mice. Moreover, the effect of opioid receptor (MOP, DOP, KOP) agonists on the mouse GI tract functions and the EOS components expression in mouse tissues and colonic biopsies from patients with functional constipation were determined. In the oldest mice, the GI peristalsis was significantly impaired as compared to the younger groups. The tissue response to MOP and DOP, but not KOP, agonists weakened with age in vitro; for DOP, it was confirmed in vivo. In the mouse upper GI tract, Oprm1, Oprd1, Oprk1 expression decreased with age; in the colon, Oprm1 expression increased. There were no differences in the expression of these genes in the colonic biopsies from patients >50 years old as compared to the younger group. In conclusion, the age-related impairment of the GI peristalsis may result from reduced MOP and DOP response to the activation with opioid agonists or the alterations in the EOS expression.     

Styryl-based compounds as potential in vivo imaging agents for beta-amyloid plaques

A group of styryl-based neutral compounds has been synthesized in this study for potential use as in vivo imaging agents for beta-amyloid plaques. Of 56 candidates, 14 compounds were found to label beta-amyloid plaques well on Alzheimer's disease (AD) human brain sections in vitro. The binding affinity to beta-amyloid fibrils was then determined by measuring the change in fluorescence intensity. Interestingly, we found that a class of quinaldine-styryl scaffold compounds displays specific binding to beta-amyloid fibrils. A representative compound, STB-8, was used in ex vivo and in vivo imaging experiments on an AD transgenic mouse model and demonstrated excellent blood-brain barrier (BBB) permeability and specific staining of the AD beta-amyloid plaques.