伴IgA沉积的微小病变肾病综合征患者血清IgA1糖基化缺陷的检测

目的 通过对伴IgA沉积的微小病变肾病综合征（MCNS-IgA）患者血清中IgA1分子糖基化缺陷程度的检测,探讨MCNS-IgA的可能的病理分类归属. 方法 选择北京大学人民医院肾内科MCNS-IgA患者10例,微小病变肾病综合症（MCNS）患者10例、大量蛋白尿IgAN （H-IgAN）患者10例为对照.用双抗体夹心ELISA法检测各组患者血清IgA1的相对浓度,用黑木凝集素检测IgA1分子的α2,6唾液酸水平,花生凝集素检测IgA1分子的半乳糖水平,蜗牛凝集素检测IgA1分子的N-乙酰氨基半乳糖水平,计算经血清IgA1浓度校正的各糖基水平.观察MCNS-IgA组患者血清IgA1分子糖基化缺陷情况,并且与H-IgAN及MCNS组进行比较.结果 与MCNS组相比,MCNS-IgA肾病患者血清IgA1的α2,6唾液酸（1.232±0.250比较1.379±0.623,P=0.455）、半乳糖缺失（0.204±0.053 vs 0.229±0.088,P=0.454）水平及N-乙酰氨基半乳糖暴露（0.191±0.039 vs0.205±0.068,P=0.626）水半无明显差异.但其血清IgA1分子α2,6唾液酸缺失（1.232±0.250vs 0.756±0.243,P=0.015）及N-乙酰氨基半乳糖的暴露（0.191±0.039比0.258±0.066,P=0.025）显著低于H-IgAN组,半乳糖缺失少,但未达统计学差异（0.204±0.053比0.139±0.038,P=0.052）. 结论 MCNS-IgA组患者血清IgA1糖基化水平上显示了与IgA肾病不同的特点,提示其可能是微小病变伴IgA分子的非特异性沉积.     

The comparison of animal models for premature ovarian failure established by several different source of inducers

The objective of this study was to compare premature ovarian failure animal models established by several different source of inducers. Female ICR mice, KM mice, and SD rats were treated by cyclophosphamide at 120 mg/kg, busulfan at 12 mg/kg, cisplatin at 3 or 4 mg/kg, 4-vinylcyclohexene diepoxide at 160 mg/kg, 35% galactose food pellet, and tripterygium glycosides at 50 mg/kg, respectively. Parameters were analyzed by body weight, serum concentration level of related hormones, ovarian and uterine pathological examination. The results indicated the body weight of mice increased very slowly following single dose of cyclophosphamide (p < 0.05) with damaged ovary; repeated doses of cisplatin could induce body weight significantly decreased (p < 0.01) with a rising trend of serum LH concentration, declining tendency of serum E2 concentration and injured ovary and uterus; 4-vinylcyclohexene diepoxide also hindered the mice growing (p < 0.05) with damaged ovary and uterus; the body weight of mice feed by 35% galactose food pellet increased slowly (p < 0.05) with dramatically higher serum concentration level of galactose, albumin, and total protein (p < 0.001) and injured ovary. Busulfan and tripterygium glycosides did not present obvious evidences. In conclusion, the inducers presented their respective features in such animal models and should be appropriately applied in preventive methods.     

The effect of dicoumarol and aesculine on the accumulation ofD-galactose by the intestinal rings of hamsters

Summary The accumulation ofD-galactose by rings of hamster small intestine shows a Michaelis-Menten kinetics with an apparent K_m of 5.5 mM and a maximum rate of net penetration of 0.35 micromoles/100 mg tissue w.w./min. Dicoumarol when present in the medium at concentrations 10^–5 to 10^–4 M, markedly inhibits (40–72%) galactose accumulation, behaving as a noncompetitive inhibitor with an apparent K_i of 5.5×10^–5 M. The drug also inhibits the oxygen uptake by the tissue in the absence of added substrate.Inhibition of galactose active transport by dicoumarol was also observed if the incubation was carried out after a preincubation with the drug. This effect could not be eliminated by repeated washings of the rings after preincubation.The effects of dicoumarol are explained as due to binding to cellular constituents and to inhibition of cell metabolism. Aesculine up to 10^–4 M concentration does not affect galactose accumulation.     

肿瘤浸润性淋巴细胞结合偶联半乳糖抗CD3单克隆抗体的肝脏靶向性研究

目的　探讨结合偶联半乳糖（ Ｇａｌａｃｔｏｓｅ , Ｇａｌ） 抗 Ｃ Ｄ３ 单克隆抗体（ Ａｎｔｉ Ｃ Ｄ３ Ｍｃ Ａｂ） 肿瘤浸润性淋巴细胞（ Ｔｕｍｏｒｉｎｆｉｌｔｒａｔｉｎｇ ｌｙｍｐｈｏｃｙｔｅｓ , Ｔ Ｉ Ｌ） 肝脏靶向性。 方法　本组把大鼠抗小鼠 Ｃ Ｄ 单克隆抗体和半乳糖偶联在一起,与３ Ｈ Ｔｄ Ｒ 标记 Ｔ Ｉ Ｌ结合后,从鼠尾静脉注入小鼠体内,在注射不同时间眼眶取血０５ ｍｌ,然后处死,切除肝、脾、肺,分别称重后进行放射性强度测定。 结果　结果显示：注射 Ｇａｌａｎｔｉ Ｃ Ｄ３ Ｍｃ Ａｂ Ｔ Ｉ Ｌ小鼠肝脏比注射单纯 Ｔ Ｉ Ｌ 的小鼠肝脏具有较高的放射性（ Ｐ＜ ００１） ,且该放射性持续较长一段时间,而脾、肺、血液内放射性强度差异无显著意义（ Ｐ＞ ００５ , Ｐ＞ ００５ , Ｐ＞ ００５） 。结论　该结果提示： Ｇａｌａｎｔｉ Ｃ Ｄ３ Ｍｃ Ａｂ Ｔ Ｉ Ｌ较单纯 Ｔ Ｉ Ｌ具有更好的肝脏靶向性。     

半乳糖基壳聚糖5氟尿嘧啶纳米粒的制备及其抑制小鼠结肠癌的实验研究

目的:半乳糖基壳聚糖纳米材料与5-氟尿嘧啶(5-FU)合成5-FU纳米粒,并观察其在体内抑制小鼠结肠癌的疗效.方法:将体外培养的C26细胞接种小鼠肝左叶,成瘤后,分别用0.9％氯化钠液、空纳米粒、5-FU和5-FU纳米粒行尾静脉注射,观察5-FU纳米粒的抗肿瘤效果.用酶联免疫吸附试验法检测血清γ干扰素(IFN-γ)和白细胞介素-2(IL-2)含量,用四甲基偶氮唑盐(MTT)比色法测定小鼠脾自然杀伤细胞(NK细胞)和细胞毒性T细胞(CTL细胞)活性.结果:与0.9％氯化钠液组及纳米粒组相比,5-FU组和5-FU纳米粒组的瘤质量显著降低(P＜0.01),尤其5-FU纳米粒组降低更显著(P＜0.01).与其他各组比较,5-FU组小鼠,外周血IFN-γ和IL-2含量均显著降低(P＜0.01),5-FU纳米粒组与0.9％氯化钠液组比较差异无统计学意义(P＞0.05).5-FU组的脾NK细胞和CTL细胞杀伤活性较其他各组显著降低(P＜0.01),5-FU纳米粒组与0.9％氯化钠液组比较未见降低(P＞0.05),纳米粒组NK和CTL杀伤活性0.9％氯化钠液组增高(P＜0.05).结论:5-FU纳米粒治疗结肠癌模型,具有明显抗肿瘤效应,纳米粒材料具有提高机体免疫功能,减低5-FU对机体免疫的抑制作用.     

Haemaphysalis longicornis tick bites are a possible cause of red meat allergy in Japan

Recent studies revealed that Amblyomma or Ixodes tick bites may cause red meat allergy, in which galactose‐α‐1,3‐galactose (α‐Gal) is a major IgE‐binding epitope. The incidence of red meat allergy is high in Shimane Prefecture, as is tick‐transmitted Japanese spotted fever. Therefore, we speculated that tick bites may cause these meat allergies. The carbohydrate α‐Gal was detected in the salivary gland protein of Haemaphysalis longicornis (H. longicornis), the vector for Japanese spotted fever, by immunoblotting using anti‐α‐Gal antibody. H. longicornis salivary gland protein‐specific IgE was detected in the sera of 24 of 30 patients with red meat allergies. Sensitization to tick salivary gland protein containing α‐Gal is possibly a major etiology of red meat allergy; the carbohydrate plays a crucial role in its allergenicity. These results further indicate that the α‐Gal epitope is present not only in Amblyomma or Ixodes, but also in Haemaphysalis.     

Toll样受体4在D-氨基半乳糖/内毒素介导的急性肝损伤中的表达

目的研究D-氨基半乳糖(D-Gal)/内毒素介导的急性肝损伤模型中肝组织细胞Toll样受体4(TLR4)的表达变化,探讨TLR4在识别内毒素、启动炎性肝损伤中的作用.方法 BALB/C小鼠腹内注射D-Gal 900 mg/kg与内毒素(即脂多糖,LPS)10 μg/kg后0～7 h分别检测血清丙氨酸转氨酶(ALT)及天门冬氨酸转氨酶(AST)与血浆肿瘤坏死因子(TNF-α)含量以评价肝功能;另随机取10只小鼠给药后观察致死率.用半定量逆转录-聚合酶链反应(RT-PCR)和Tanon Gis 2.0软件分析不同时间点肝组织中TLR4 mRNA表达,并与血浆TNF-α含量进行相关分析;免疫组织化学观察肝组织TLR4蛋白的表达.实验数据用SAS软件分析.结果给药后4 h,血清转氨酶明显升高(与0h比较,P＜0.05);血浆TNF-α含量逐步上升,在2 h、5 h有两个分泌高峰;7 h小鼠开始死亡,10 h致死率达80%.正常小鼠肝组织少量表达TLR4 mRNA,给药后表达明显增强(与0 h比较,P＜0.05);免疫组织化学也显示TLR4蛋白有类似的变化,尤其肝窦内皮细胞、库普弗细胞表达更为显著.血浆TNF-α含量与肝组织TLR4 mRNA表达呈正相关.结论肝内细胞膜上表达的TLR4可能通过启动下游的炎性应答基因表达及细胞因子释放而诱导出肝组织对LPS的炎性应答.因此,调控TLR4表达可能是感染性疾病防治的一种新策略.     

The functioning liver mass

The liver function results from the interaction of anatomical, histological and biochemical aspects. Thus the concept of functioning liver mass must be related to the many biological aspects involved in the process under evaluation, and its measure can be viewed as a virtual parameter averaging the liver function. In the present review the functioning liver mass has been categorized with respect to the many aspects involved (biochemical transformations, membrane transports and passive intraluminal transports) and their interactions (intracellular, transcellular, transacinar and flow-dependent). Parameters reflecting the functioning liver mass (whether static or dynamic) have been considered with respect to the experimental conditions and characterized according to the type of estimate they provide (capacity or activity). Methods for evaluating the functioning liver mass have been presented and discussed, and a protocol has been proposed for clinical applications, based on the association of the galactose elimination capacity measuring the metabolic mass with the hepatic clearance of D-sorbitol evaluating the functional hepatic plasma flow. This work was supported in part by grants from the Ministero della Pubblica Istruzione, Roma, Italy and the Regione Piemonte.     

肝靶向性Gal-PEG-PEI纳米载体介导psiRNA转染HepG2细胞效率的检测

目的 检测肝细胞靶向性Gal-PEG-PEI纳米基因载体介导psiRNA转染HepG2细胞的效率.方法 纳米粒径仪和透射电镜测定Gal-PEG-PEI/psiRNA复合物的粒径和形态,转染HepG2细胞,48 h后用流式细胞仪测定转染效率.转染前加入1 mg半乳糖观察其半乳糖竞争拮抗结果.结果 Gal-PEG-PEI/psiRNA复合物的粒径随N/P的增大而减小,最小粒径为81.1 nm;Gal-PEG-PEI载体的转染效率为(37.34±2.50)%,明显高于PEG-PEI、LipofectamineTM 2000及裸psiR-NA;加入竞争拮抗剂半乳糖后Gal-PEG-PEI的转染效率下降至3.60%.结论 Gal-PEG-PEI纳米基因载体对HepG2细胞有较高的转染效率,具有良好的肝细胞靶向性.     

升降散加减方治疗IgAN大鼠作用机制探讨

目的 通过探讨疏通三焦、清热利湿理论指导下的升降散加减方对IgA肾病(immunoglobulin a nephropathy,IgAN)大鼠血清转化生长因子 β1(transforming growth factorβ1,TGF-β1)、白细胞介素-6(interleukin-6,IL-6)、半乳糖缺乏-IgA(ga...