重组人血小板生成素在小鼠体内的代谢分布和排泄

用125Ⅰ标记结合分子排阻高效液相色谱(SHPLC)研究小鼠体内重组人血小板生成素(rhTPO)代谢、分布和排泄.标记-rhTPO体外可刺激TPO依赖细胞系TD3增殖;体内刺激外周血血小板增加活性与未标记标准rhTPO相似,而SHPLC放化纯度(96.9±1.5)%(n=3)符合药动学研究要求.小鼠皮下注射1微克/鼠后血清和尿SHPLC分析除原形外,出现了分子量较小的125Ⅰ-代谢物Ⅰ和Ⅱ.血清以原形和125Ⅰ-代谢物Ⅰ为主,尿中有少量原形,主要是125Ⅰ-代谢物Ⅰ和Ⅱ.原形消除半衰期为10.8小时,组织TCA可沉淀放射性分布特点是靶器官骨髓较高,泌尿系统最高,脑内最低.主要经尿排泄,少量经粪排泄.72小时排出(98.3±5.6)%.     

血红素加氧酶-1在铁代谢中的作用

铁是机体必需的微量元素,有着广泛的生物学作用,包括氧化运输和细胞呼吸作用,缺铁会引起细胞生长停滞或死亡,铁过载会使细胞发生氧化应激进而损伤细胞膜、蛋白质、核酸.血红素加氧酶-1(HO-1)分解血红素产生铁是铁重复利用最主要的来源,在铁代谢中的作用至关重要.     

糖皮质激素治疗SLE对骨代谢标志物的影响分析

目的 研究骨代谢标志物在糖皮质激素治疗系统性红斑狼疮(SLE)前后的变化,及其与治疗疗程进展的关系,探讨激素治疗后育龄期、围绝经期、绝经后女性患者骨代谢水平的特征;分析骨代谢标志物与骨密度在糖皮质激素治疗SLE中诊断骨质疏松的价值.方法 检测40例SLE患者经糖皮质激素治疗前及治疗后3、6、12个月的骨代谢标志物(BGP、β-CTX、PICP),测定不同部位的骨密度,并运用统计学方法进行处理.结果 ①糖皮质激素治疗前骨形成标志物和骨吸收标志物均较正常对照组明显降低(P＜0.05),差异有统计学意义;②激素治疗早期,骨形成标志物BGP、PICP明显下降,骨吸收标志物β-CTX明显上升,治疗3、6、12个月后的骨代谢标志物水平与治疗前比较差异有统计学意义(P＜0.05);③激素治疗后,围绝经期组较育龄期组β-cTx、PTH、FSH水平明显升高(P＜0.05),绝经后组与围绝经期组比较,β-CTX、BGP、PICP、FSH均显著升高(P＜0.05),差异有统计学意义.④糖皮质激素治疗3、6、12个月后测股骨近端、腰椎、Ward三角的骨密度,其不同部位骨密度的变化差异缺乏一致性.结论 糖皮质激素治疗SLE患者,骨代谢标志物在治疗早期有明显变化;而同阶段不同部位骨密度变化存在差异,骨代谢标志物对早期诊断骨质疏松有重要意义,骨代谢标志物联合雌二醇(E2)可为围绝经女性骨质流失的评估提供参考.     

Mapping human brain capillary water lifetime: high‐resolution metabolic neuroimaging

Shutter‐speed analysis of dynamic‐contrast‐agent (CA)‐enhanced normal, multiple sclerosis (MS), and glioblastoma (GBM) human brain data gives the mean capillary water molecule lifetime (τ_b) and blood volume fraction (v_b; capillary density–volume product (ρ^?V)) in a high‐resolution ¹H₂O MRI voxel (40 μL) or ROI. The equilibrium water extravasation rate constant, k_po (τ_b^?1), averages 3.2 and 2.9 s^?1 in resting‐state normal white matter (NWM) and gray matter (NGM), respectively (n = 6). The results (italicized) lead to three major conclusions. (A) k_po differences are dominated by capillary water permeability (P_W^?), not size, differences. NWM and NGM voxel k_po and v_b values are independent. Quantitative analyses of concomitant population‐averaged k_po, v_b variations in normal and normal‐appearing MS brain ROIs confirm P_W^? dominance. (B) P_W^? is dominated (>95%) by a trans(endothelial)cellular pathway, not the P_CA^? paracellular route. In MS lesions and GBM tumors, P_CA^? increases but P_W^? decreases. (C) k_po tracks steady‐state ATP production/consumption flux per capillary. In normal, MS, and GBM brain, regional k_po correlates with literature MRSI ATP (positively) and Na⁺ (negatively) tissue concentrations. This suggests that the P_W^? pathway is metabolically active. Excellent agreement of the relative NGM/NWM k_pov_b product ratio with the literature ³¹PMRSI‐MT CMR_oxphos ratio confirms the flux property. We have previously shown that the cellular water molecule efflux rate constant (k_io) is proportional to plasma membrane P‐type ATPase turnover, likely due to active trans‐membrane water cycling. With synaptic proximities and synergistic metabolic cooperativities, polar brain endothelial, neuroglial, and neuronal cells form “gliovascular units.” We hypothesize that a chain of water cycling processes transmits brain metabolic activity to k_po, letting it report neurogliovascular unit Na⁺,K⁺‐ATPase activity. Cerebral k_po maps represent metabolic (functional) neuroimages. The NGM 2.9 s^?1 k_po means an equilibrium unidirectional water efflux of ~10¹⁵ H₂O molecules s^?1 per capillary (in 1 μL tissue): consistent with the known ATP consumption rate and water co‐transporting membrane symporter stoichiometries. © 2015 The Authors NMR in Biomedicine Published by John Wiley & Sons Ltd.     

Narcolepsy and pregnancy: a retrospective European evaluation of 249 pregnancies

In a retrospective cohort study undertaken in 12 European countries, 249 female narcoleptic patients with cataplexy (n = 216) and without cataplexy (n = 33) completed a self‐administrated questionnaire regarding pregnancy and childbirth. The cohort was divided further into patients whose symptoms of narcolepsy started before or during pregnancy (308 pregnancies) and those in whom the first symptoms of narcolepsy appeared after delivery (106 pregnancies). Patients with narcolepsy during pregnancy were older during their first pregnancy (P < 0.001) and had a higher body mass index (BMI) prior to pregnancy (P < 0.01). Weight gain during pregnancy was higher in narcoleptic patients with cataplexy (P < 0.01). More patients with narcolepsy–cataplexy during pregnancy had impaired glucose metabolism and anaemia. Three patients experienced cataplexy during delivery. The rate of caesarean sections was higher in the narcolepsy–cataplexy group compared to the narcolepsy group (P < 0.05). The mean birth weight and gestational age of neonates were within the normal range and did not differ across groups. Neonatal care was affected adversely by symptoms of narcolepsy in 60.1% of those with narcolepsy during pregnancy. This study reports more obstetric complications in patients with narcolepsy–cataplexy during pregnancy; however, these were not severe. This group also had a higher BMI and higher incidence of impaired glucose metabolism during pregnancy. Caesarian section was conducted more frequently in narcolepsy–cataplexy patients, despite cataplexy being a rare event during delivery. Furthermore, symptoms of narcolepsy may render care of the infant more difficult.     

我国弓形虫Chinese 1优势基因型感染对小鼠脑组织铁代谢影响

目的　探讨我国弓形虫Chinese 1优势基因型感染对宿主脑组织铁代谢及脑损伤的影响。方法　将20只C57BL/6（体质量15～17 g）小鼠随机分为对照组和感染组,每组10只。感染组每只小鼠腹腔注射4 000个弓形虫Chinese 1优势基因型TgCtwh3虫株速殖子,对照组小鼠注射等量无菌PBS,饲养6 d后处死小鼠并取其脑组织。采用电感耦合等离子体质谱法（inductively coupled plasma mass spectrometry, ICP⁃MS）检测小鼠脑组织铁元素水平;采用RNA芯片检测两组小鼠脑组织差异基因数目并对功能基因表达情况进行基因本体论（Gene Ontology, GO）功能富集;采用实时荧光定量PCR（fluorescent quantitative real⁃time PCR, qPCR）技术检测小鼠脑组织中弓形虫表面抗原1（Toxoplasma gondii surface antigen 1,TgSAG1）基因及部分锌铁调控蛋白（Zrt⁃ and Irt⁃like protein, ZIP）家族mRNA表达水平;采用光镜和电镜观察小鼠脑组织海马齿轮回（dentate gyrus, DG）超微结构;采用Western blotting检测谷胱甘肽过氧化物酶4（glutathione peroxidase 4, GPx4）蛋白表达水平;采用硫代巴比妥酸（TBA）法检测丙二醛（malondialdehyde, MDA）水平;采用免疫组化检测血管内皮生长因子（vascular endothelial growth factor, VEGF）蛋白表达光密度（optical density, OD）值。结果　光镜下可见感染组小鼠脑组织海马DG区细胞坏死,电镜下见感染组小鼠脑组织海马区出现胞质空泡化、核皱缩坏死、线粒体嵴断裂消融、自噬小体增加等超微结构变化。与对照组相比,感染组小鼠脑组织中铁元素水平上调[（32.92 ± 0.90） µg/g vs.（37.72 ± 1.10） µg/g;t = 3.397, P < 0.01];RNA芯片检测感染组小鼠脑组织发现721个基因上调、276个基因下调,差异表达基因在金属离子结合能力上有明显富集。与对照组相比,感染组小鼠脑组织金属元素转运体ZIP2 mRNA表达水平上调（t = 8.659,P < 0.05）、GPx4表达下降[（1.046 ± 0.025） vs. （0.720 ± 0.101）;t = 3.129,P < 0.01]）、MDA水平升高[（4.37 ± 0.33） nmol/mgprot vs.（5.93 ± 0.54） nmol/mgprot;t = 2.451,P < 0.05）]、VEGF蛋白平均OD值上调[（0.348 3 ± 0.017 8） vs. （0.490 6 ± 0.010 5）;t = 6.641,P < 0.01]。结论　Chinese 1优势基因型弓形虫感染后,小鼠脑组织中铁元素蓄积、抗氧化能力下调、氧化应激水平升高,提示弓形虫感染可影响宿主脑组织铁代谢而导致脑损伤。     

渐进性咬合紊乱致大鼠咬肌超微结构损伤及能量代谢变化的研究

目的：通过观察经历不同时长实验性渐进性咬合紊乱后大鼠咬肌的超微结构及肌酸激酶（CK）和乳酸脱氢酶（LDH）含量变化的不同,以探讨渐进性咬合紊乱对大鼠咬肌的影响程度与时间之间的关系。方法：取8周龄SPF级雄性SD大鼠随机等分为2、4、6、8周4组,每个实验组各有一个空白对照组,每组8只,空白对照组不给予任何处理,实验组通过二次分牙操作,建立渐进性咬合紊乱模型,并分别于放入最后一个皮圈后2、4、6、8周处死,HE染色和透射电镜观察大鼠咬肌超微结构,同时采用酶联免疫法（ELISA）检测各大鼠咬肌中乳酸脱氢酶（LDH）及肌酸激酶（CK）的含量,以探讨咬合紊乱对咬肌能量代谢活动的影响,进一步增加对渐进性咬合紊乱与口颌系统活动关系的认识。结果：2、4、6、8周的渐进性咬合紊乱后,大鼠咬肌线粒体均出现损伤,且随时间的延长,损伤加重,CK及LDH的活性也逐渐增加（P＜0．05）。结论：渐进性咬合紊乱对大鼠咬肌损伤与作用时间成正相关。     

肝组织CTGF、TGF-β_1的表达与肝纤维化关系的实验研究

目的:探讨免疫性肝纤维化肝组织结缔组织生长因子(CTGF)与转化生长因子β1(TGF-β1)的表达与肝纤维化的关系。方法:用猪血清大鼠腹腔内注射复制免疫损伤性肝纤维化模型,免疫攻击16周后处死动物,行肝组织病理分析,免疫组化法检测肝内CTGF、TGF-β1表达。结果:16周后模型组大鼠形成典型的肝纤维化,肝内CTGF与TGF-β1表达均显著增强,同时二者阳性分布部位相近,表达程度呈正相关性;且CTGF表达与组织病理上肝纤维化变化呈正相关性。结论:实验性肝纤维化过程中CTGF与TGF-β1促进肝纤维化的进展。     

不同麻醉下电针对脑缺血再灌注大鼠脑能量代谢及氧供需平衡的影响

目的观察脑缺血再灌注损伤后大鼠颈静脉血糖、乳酸和血氧饱和度的变化,以及不同麻醉下电针对大鼠颈静脉血糖、乳酸和血氧饱和度的影响。方法雄性SD大鼠40只,随机分为5组,即:假手术组(SH)、水合氯醛+脑缺血再灌注组(CL+CI/R)、水合氯醛+脑缺血再灌注+电针组(CL+CI/R+EA)、异丙酚+脑缺血再灌注组(P+CI/R)、异丙酚+脑缺血再灌注+电针组(P+CI/R+EA),每组8只。采用4-VO法建立大鼠全脑缺血模型,于再灌流开始后,CI/R+EA组电针"百会""命门"和"足三里"穴,电针参数:频率30~50 Hz,间断疏密波,电流强度1 mA,以局部肌肉轻微震颤为度,时间20 min。在缺血后24 h,从右颈静脉抽取0.5 mL血样,进行血糖、乳酸和血氧饱和度的测定。结果CL+CI/R组大鼠的颈静脉血糖和血氧饱和度显著高于SH组(P0.01),乳酸值则低于SH组(P0.01),P+CI/R组大鼠的颈静脉血糖和血氧饱和度明显高于SH组(P0.05),乳酸值则显著低于SH组(P0.01);与CL+CI/R组相比,CL+CI/R+EA组和P+CI/R+EA组的颈静脉血糖和血氧饱和度显著降低(P0.01),而乳酸值则显著升高(P0.01);与P+CI/R组相比,实施电针治疗的P+CI/R+EA组的颈静脉血糖显著降低(P0.01),而乳酸值则明显升高(P0.05);P+CI/R组的颈静脉血糖明显低于CL+CI/R组的血糖(P0.05)。结论针刺治疗可明显改善脑缺血再灌注大鼠的糖代谢,增加脑细胞对葡萄糖和氧的摄取和利用,不同的静脉麻醉药物对电针治疗效果的影响无显著差异性。     

甘精胰岛素和二甲双胍联合治疗对初诊2型糖尿病患者炎症因子的影响

目的 观察甘精胰岛素与二甲双胍联合治疗对初诊2型糖尿病(T2DM)患者炎症因子的影响.方法 对110例新诊断的T2DM患者进行甘精胰岛素(起始剂量10 U/d)与二甲双胍(0.5 g,每天3次)联合治疗(T2DM组),疗程12周.另入选100例同期进行健康体检的正常人群为正常对照组.观察2组研究对象基线血糖指标空腹血糖(FPG)、餐后2 h血糖(2 hPG)和糖化血红蛋白(HbA_1c)以及炎症因子C-反应蛋白(CRP)、肿瘤坏死因子α(TNF-α)和白细胞介素-6(IL-6)的差异,并比较T2DM组治疗前后血糖指标和炎症因子的变化.结果 观察前T2DM组的血糖指标及炎症因子明显高于对照组(P均＜0.05),其他基础临床资料没有差异(P均＞0.05).经甘精胰岛素和二甲双胍联合治疗12周后,T2DM组的血糖指标明显改善,FPG:治疗前(14.8±3.9)mmoL/L,治疗后(6.6±2.1)mmol/L;2 hPG:治疗前(17.6±3.3)mmol/L,治疗后(8.3±1.2)mmoL/L;HbA1c:治疗前(9.6±2.7)%,治疗后(6.5±0.8)%(t值分别为7.40、8.37、3.98,P均＜0.01).炎症因子水平也显著下降,CRP:治疗前(8.8±2.5)mg/L,治疗后(5.5±1.4)mg/L;TNF-α:治疗前(2.9±0.6)ng/L,治疗后(1.6±0.2)ng/L;IL-6:治疗前(170.3±22.2)pg/L,治疗后(105.9±14.6)pg/L(t值分别为4.61、3.52、5.68,P均＜0.05).结论 甘精胰岛素与二甲双胍联合治疗可以改善初诊T2DM患者的糖代谢,降低患者炎症因子的水平.