In Vitro Anticancer and Cancer-Preventive Activity of New Triterpene Glycosides from the Far Eastern Starfish Solaster pacificus

Sea stars or starfish (class Asteroidea) and holothurians or sea cucumbers (class Holothuroidea), belonging to the phylum Echinodermata (echinoderms), are characterized by different sets of glycosidic metabolites: the steroid type in starfish and the triterpene type in holothurians. However, herein we report the isolation of eight new triterpene glycosides, pacificusosides D–K (1–3, 5–9) along with the known cucumarioside D (4), from the alcoholic extract of the Far Eastern starfish Solaster pacificus. The isolated new compounds are closely related to the metabolites of sea cucumbers, and their structures of 1–3 and 5–9 were determined by extensive NMR and ESIMS techniques. Compounds 2, 5, and 8 have a new type of tetrasaccharide chain with a terminal non-methylated monosaccharide unit. Compounds 3, 6, and 9 contain another new type of tetrasaccharide chain, having 6-O-SO₃-Glc as one of the sugar units. The cytotoxic activity of 1–9 against non-cancerous mouse epidermal cells JB6 Cl41 and human melanoma cell lines SK-MEL-2, SK-MEL-28, and RPMI-7951 was determined by MTS assay. Compounds 1, 3, 4, 6, and 9 showed potent cytotoxicity against these cell lines, but the cancer selectivity (SI > 9) was observed only against the SK-MEL-2 cell line. Compounds 1, 3, 4, 6, and 9 at the non-toxic concentration of 0.1 μM significantly inhibited neoplastic cell transformation of JB6 Cl41 cells induced by chemical carcinogens (EGF, TPA) or ionizing radiation (X-rays and UVB). Moreover, compounds 1 and 4 at the non-toxic concentration of 0.1 µM possessed the highest inhibiting activity on colony formation among the investigated compounds and decreased the colonies number of SK-MEL-2 cells by 64% and 70%, respectively. Thus, triterpene glycosides 1 and 4 can be considered as prospective cancer-preventive and anticancer-compound leaders.     

In Vitro and In Vivo Dendritic Cell Immune Stimulation Effect of Low Molecular Weight Fucoidan from New Zealand Undaria pinnatifida

Low molecular weight fucoidan (LMWF) has been reported to have immunomodulation effects through the increase of the activation and function of macrophages. In this study, the regulating effect of LMWF from Undaria pinnatifida grown in New Zealand on dendritic cells (DCs) was investigated. We discovered that LMWF could stimulate DCs’ maturation and migration, as well as CD4+ and CD8+ T cells’ proliferation in vitro. We proved that this immune promoting activity is activated through TLR4 and its downstream MAPK and NF–κB signaling pathways. Further in vivo (mouse model) investigation showed that LMWF has a strong immunological boosting effect, such as facilitating the proliferation of immune cells and increasing the index of immune organs. These findings suggest that LMWF has a positive immunomodulatory effect and is a promising candidate to supplement cancer immunotherapy.     

Functional changes of retinal ganglion cells in rd1 mice during midterm of retinal pigmentosa

AIM: To investigate how the function of retinal ganglion cells (RGCs) change in the midterm of retinal pigmentosa (RP) in rd1 mice (a transgenic animal model of RP). METHODS: The action potentials from multiple RGCs in rd1 mice at postnatal 20 d (P20) or normal C57 mice (control) were simultaneously recorded by multi-electrode array recording. The functional changes of surviving ganglion cells were evaluated by comparing spontaneous and light-evoked activities of RGCs between rd1 and control mice. The extent of photoreceptor degeneration was verified by immunohistochemical staining. RESULTS: Immunohistochemistry results showed the thickness of the retinal photoreceptor layer of rd1 mice was significantly lower than that in normal mice at P20. According to the light response properties, we classified ganglion cells into 6 subgroups: ON sustained, ON transient, ON-OFF sustained, ON-OFF transient, OFF sustained and OFF transient RGCs, with a very tiny percentage of OFF sustained RGCs (1.0%~3.1%). The percentage of RGCs remaining light responsive in rd1 mice was significantly lower than that in C57 mice. The average spontaneous spiking rate for rd1 RGCs was overall significantly increased compared to that in C57 cells, whereas different RGC types had different changes. The light-induced responses and light sensitivities of all types of RGCs in rd1 mice were both significantly lower than those in C57 mice. CONCLUSION: The photoreceptors of rd1 mice are severely degenerated in the midterm of retinal degeneration. The functions of RGCs in rd1 mice in the midterm of degeneration decay obviously, with variance in different RGCs types.     

含蛋氨酸二肽影响奶牛乳腺上皮细胞乳蛋白合成相关基因表达

本试验旨在研究含蛋氨酸(Met)二肽对奶牛乳腺上皮细胞(BMECs)内乳蛋白合成相关基因表达的影响。试验分3部分,均采用单因子完全随机试验设计,Met的添加浓度及培养时间分别为60μg/m L(0.402 mmol/L)、48 h。第1部分,培养液添加8种含Met二肽[蛋氨酸-蛋氨酸(P-Met-Met)、蛋氨酸-赖氨酸(P-Met-Lys)、蛋氨酸-色氨酸(P-Met-Trp)、蛋氨酸-苯丙氨酸(P-Met-Phe)、蛋氨酸-苏氨酸(P-Met-Thr)、蛋氨酸-异亮氨酸(P-Met-Ile)、蛋氨酸-亮氨酸(P-Met-Leu)、蛋氨酸-缬氨酸(P-Met-Val)],以不添加二肽为对照,测定BMECs乳蛋白合成相关基因(αs1-酪蛋白、β-酪蛋白、κ-酪蛋白、β-乳球蛋白、Ⅱ型小肽转运载体和氨肽酶氮)的表达量;第2部分,培养液添加8种与上述二肽对应的游离氨基酸(F-Met-Met、F-Met-Lys、F-MetTrp、F-M et-Phe、F-M et-Thr、F-M et-Ile、F-M et-Leu、F-M et-Val),以不添加游离氨基酸为对照,测定BM ECs乳蛋白合成相关基因的表达量;第3部分,用二肽等物质的量替代相应游离氨基酸,测定BMECs乳蛋白合成相关基因的表达量以及细胞内外氨肽酶含量。结果表明:P-Met-Met和P-M et-Lys组较对照组和其他二肽组上调了αs1-酪蛋白和β-酪蛋白基因的表达量,且P-M et-M et组优于P-Met-Lys组。F-Met-Met和F-Met-Lys组较对照组和其他游离氨基酸组显著提高了αs1-酪蛋白基因的表达量(P0.05)。除P-Met-Val和P-Met-Leu组外,其他二肽替代游离氨基酸后均不同程度地提高了乳蛋白和Ⅱ型小肽转运载体基因的表达量,其中P-Met-Met表现出较好的促进效果。总之,含Met二肽等量替代对应的游离氨基酸能够促进乳蛋白基因的表达,其中尤以P-Met-Met的效果最好。     

颈静脉灌注精氨酸对泌乳中期奶牛血清生化和免疫指标的影响

本文旨在研究颈静脉灌注精氨酸对泌乳中期奶牛血清生化和免疫指标的影响。采用3×3复式拉丁方试验设计,选择6头体重、胎次、泌乳期、泌乳量和体况基本一致的荷斯坦奶牛为试验动物,随机分为3组(每组2头),分别为酪蛋白组(对照组)、精氨酸组(灌注37.66 g/d)、丙氨酸组(与精氨酸组等氮,灌注77.24 g/d);每个试验期为22 d(7 d灌注+15 d间隔),在每个试验灌注期的最后1 d晨饲前采血,测定血清生化和免疫指标。结果表明:精氨酸组的奶牛血清中总蛋白、免疫球蛋白G(Ig G)、肿瘤坏死因子α(TNF-α)浓度要显著高于其他2组(P0.05),白球比、白细胞介素1(IL-1)浓度显著高于丙氨酸组(P0.05),但是牛奶中体细胞数显著低于其他2组(P0.05),而血清中谷丙转氨酶、谷草转氨酶活性,免疫球蛋白A(Ig A)、免疫球蛋白E(Ig E)、白细胞介素6(IL-6)浓度在组间无显著差异(P0.05)。综上,灌注精氨酸能够通过提高血清总蛋白、Ig G及IL-1、TNF-α的浓度,从而一定程度上增强泌乳奶牛机体的免疫力和胎儿的被动免疫力。     

鹿茸发育的组织来源及其相互作用机制研究进展

本综述介绍了鹿茸发育的组织来源及其相互作用机制的研究进展,阐明了鹿茸发育的组织细胞特性,进一步分析了鹿茸发育机制中待解决的问题。     

利用乳汁体细胞研究共轭亚油酸钙对奶牛乳腺基因表达的作用

8头泌乳中期的奶牛被随机分为2组，分别是对照组（每头牛：基础日粮＋棕榈酸钙200g／d）和试验组（每头牛：基础日粮＋共轭亚油酸钙200g／d），试验期14d，检测了奶产量、乳成分，分析奶牛的血液变化，并采用荧光定量PCR对乳汁体细胞中脂蛋白脂酶（LPL）、乙酰辅酶A羧化酶（ACACA）的基因表达进行检测。结果显示，与对照组相比，试验组奶牛的乳产量、乳蛋白、乳糖和乳汁体细胞含量没有显著影响，但是显著降低了乳脂肪含量（P〈0．05），对照组和试验组奶牛乳脂肪含量分别为0．0326g／mL和0．0244g／mL。检测血液指标发现，共轭亚油酸钙显著升高了奶牛血液中高密度脂蛋白的含量（P〈0．05）。基因分析发现，试验组奶牛乳汁体细胞中LPL、ACA—CA的基因表达显著下调，表明共轭亚油酸钙抑制了奶牛乳腺细胞脂肪酸合成酶的基因表达。     

The use of dialysis cells for investigating pore water composition in wetland substrata,with particular reference to dissolved iron and sulphide

Abstract

Details and some evaluation are given of the use of a dialysis method for sampling pore water from depth in wetland substrata (peats), with particular reference to measurement of concentrations of ion species with stabilities dependent on redox potential. The method is based upon the burial and subsequent retrieval (after equilibration) of cells made from dialysis membrane filled with deionized water. Preliminary results of field investigations of concentrations of dissolved iron and sulphide in the pore water of base‐rich mires, as sampled by this method, are given. Results suggest that the method could have very considerable application and that its potential, and possible problems, deserve further examination.     

Sulphido-leukotriene production from peripheral leukocytes and skin in clinically normal dogs and house dust mite positive atopic dogs

Pathogenesis of canine atopy has not been completely elucidated. In humans, sulphido-leukotrienes (s-LT) play a role in atopy, and increased production of s-LT occurs in the skin and peripheral leukocytes after allergen challenge. The study population included 16 clinically normal and 13 atopic dogs. All atopic dogs had in common a positive reaction (4+) to the intradermal injection of house dust mite (allergen of reference). Blood samples and skin biopsies were collected. Sulphido-LT synthesis by peripheral leukocytes after stimulation was measured, and no statistically significant difference was found between clinically normal and atopic dogs. Sulphido-LT concentrations in skin samples from stimulated and unstimulated sites were measured, and no statistically significant difference was detected between clinically normal and atopic dogs or between lesional and nonlesional skin within the atopic group. Clinical signs of atopic dogs were graded by owners and no correlation was found between their severity and cutaneous concentrations of s-LT. In this study there was no increase in s-LT synthesis in atopic dogs.