Metabolic profiles of the Flos Abelmoschus manihot extract by intestinal bacteria from the normal and CKD model rats based on UPLC‐Q‐TOF/MS

Flos Abelmoschus manihot is a traditional herbal medicine widely used in clinical practice to tackle chronic kidney disease (CKD) for thousands of years. Nowadays, many studies indicate that gut bacteria are closely related to the progression of CKD and CKD‐related complications. In this study, a UPLC‐Q‐TOF/MS method coupled with the MetaboLynx™ software was established and successfully applied to investigate the metabolites and metabolic profile of Flos A. manihot extract by intestinal bacteria from normal and CKD rats. Eight parent components and eight metabolites were characterized by their protonated ions. Among these compounds, 15 were detected in the two group samples while M16 was only determined in the CKD model samples. Compared with the quercetin‐type glycosides, fewer myricetin‐type and gossypetin‐type metabolites were obtained in the two group samples. These metabolites suggested that deglycosylation and methylation are the major metabolic pathways of Flos A. manihot extract. Few differences of metabolite classes were observed in the two group samples. However, the concentrations of aglycones such as quercetin, myricetin and gossypetin in the normal samples were notably higher than those in the CKD model samples. The results are important in unravelling the pharmacological effects of A. manihot and clarifying its mechanism of action in vivo .     

大鼠肾脏汞结合金属硫蛋白的联用技术表征研究

通过凝胶色谱(SEC)及反相液相色谱(RPC)分别与电感耦合等离子体质谱(ICP-MS)和电喷雾电离质谱(ESI-MS)联用的技术，对灌喂HgCl₂大鼠的肾组织中诱导的多金属结合金属硫蛋白(MT)的结构进行表征分析。通过SEC-ICP-MS曲线上的金属信号及紫外检测的MT光谱吸收特征，可确认组织提取液中MT在色谱曲线上的位置。与对照组相比，灌喂HgCl₂大鼠肾中Hg-MT的诱导量显著增加，与此同时，Cu-MT在染毒组的诱导量也相应增加，说明MT在染汞大鼠肾脏中起着对汞的解毒和对铜的调节作用。MT样品提取物经葡聚糖凝胶(G-75)柱分离纯化并用透析法脱盐后，用细内径反相色谱柱分离，进行梯度洗脱，并与ESI-MS联用，可获得总离子流色谱图及与色谱峰对应的质谱图，通过对质谱信号的解析及参考脱金属MT构型的有关报道，对汞诱导大鼠肾组织MT的不同亚型和次亚型分子结构进行推断。结果证明，在灌喂HgCl₂大鼠的肾组织中存在着一系列汞结合的MT形态。     

Infrared Analysis of Urinary Stones,Using a Single Reflection Accessory and a KBr Pellet Transmission*

This work is a comparative study of two FTIR techniques applied to urinary stones analysis: single reflection diamond attenuated total reflection (ATR) and KBr pellet transmission (KPT). We show that the two methods allow the identification of all stone components. The ATR technique is more useful and rapid to identify the species without sample pretreatment. Nevertheless, KPT is more appropriate for components determination in urinary stones.

These techniques were applied to the study of a series of 313 calculi. The stone constituents were first identified by ATR, and in a second step, the proportion of each species present in the stone was determined by KPT in whole‐stone mixture.

The results obtained showed the presence of 11 different components classified as follows with the frequency of detection in the stones studied: calcium oxalate monohydrate (whewellite), 78.9%; carbapatite, 33.9%; calcium oxalate dihydrate (weddellite), 24%; uric acid anhydrous, 19.2%; ammonium hydrogen urate, 7%; struvite, 4.8%; cystine, 1%; ammonium sodium urate and other phosphates (amorphous carbonated calcium phosphate, brushite, whitlockite), each in less than 1%.     

N-乙酰半胱氨酸对老年患者碘海醇肾损害的保护作用

目的探讨N-乙酰半胱氨酸对老年患者碘海醇肾损害的保护作用。方法选取行增强CT检查（静脉注射碘造影剂碘海醇）的老年患者74例,其中A组26例（于造影前48h和造影后48h口服N-乙酰半胱氨酸600mg,2次/d）,B组25例（于造影前48h和造影后48h口服N-乙酰半胱氨酸600mg,1次/d）;对照组23例（于造影前48h和造影后48h口服安慰剂,1次/d）。造影前和造影后48h,检测3组患者血尿素氮（BUN）、SCr、内生肌酐清除率（CCr）,并记录对比剂肾病发生率,造影前和造影后48h以及5d时,检测24h尿蛋白（PRO）、24h尿β2-微球蛋白(β2- MG)、24h尿微量白蛋白（UmAlb）。结果 A组无CIN发生,B组发生CIN5例（20%）,C发生CIN6例（26.09%）,A组CIN发生率明显低于B、C组（均P＜0.05）。3组患者造影前各指标的差异均无统计学意义（P＞0.05）。与造影前比较,3组患者造影后48h BUN、SCr、Ccr均发生明显变化（P＜0.05）;与B组、对照组比较,A组造影后48h BUN、SCr、Ccr均发生明显变化（P＜0.05）。与造影前比较,3组患者造影后48h、5d时PRO、Umalb、和β2- MG均发生明显变化（P＜0.05）;与对照组比较,A组造影后48h及5d、B组造影后48h的PRO、Umalb、和β2- MG均发生明显变化（P＜0.05）。结论口服N-乙酰半胱氨酸600mg（2次/d）能有效预防老年患者碘海醇肾损害。     

肌球蛋白Ⅱ缺失细胞胞质分裂机制研究

哺乳动物细胞胞质分裂过程中伴随着一系列形态学改变,随着分裂沟不断收缩,形成连接两个子细胞的细胞间桥.间桥不断拉长、变细,直至断裂、生成两个子细胞.采用细胞力学和形态学测量及分析方法,通过施加肌球蛋白Ⅱ抑制剂,定量研究了NRK细胞间桥变细动力学;采用细胞免疫荧光技术,检测了早期胞质分裂肌动蛋白的分布,揭示肌球蛋白Ⅱ缺失细胞胞质分裂可能的机制.结果表明:施加肌球蛋白Ⅱ抑制剂的NRK细胞,其整体形态学和细胞间桥形态学曲线明显不同于0.3%DMSO组.根据流体力学特性和所测量的力学参数对曲线进行模拟发现,表面张力对肌球蛋白Ⅱ抑制组细胞的间桥动力学曲线轨迹影响很大.研究结果提示由细胞力学特性决定的拉普拉斯压力和细胞运动共同参与了肌球蛋白Ⅱ缺失细胞胞质分裂的调节.     

Hemostatic Efficacy of Biological Self‐Assembling Peptide Nanofibers in a Rat Kidney Model

We evaluated the hemostatic efficacy of a biological self‐assembling peptide RADA16‐I in a rat kidney injury model. Adult male rats were randomized into five groups: sham operation (no renal excision), no hemostatic agent (control), commercially available gelatin sponge (Gelfoam), 1% RADA16‐I, and 2% RADA16‐I. After left partial nephrectomy, the anesthetized animal was anticoagulated using 300 IU · kg^?1 heparin, and the topical hemostatic agent was applied to the injury. Blood loss and mean arterial pressure (MAP) were recorded. As was the case for Gelfoam, 2% RADA16‐I produced marked hemostasis versus controls (p < 0.01). Blood loss with 1% and 2% RADA16‐I was significantly less than controls. The decline in MAP during surgery was less with 2% versus 1% RADA16‐I. RADA16‐I also resulted in less histological tissue responses than Gelfoam. These data suggest that RADA16‐I can stop hemorrhage, with only minimal tissue responses, in experimental renal injury.

     

肾脏冷灌注过程中的生物传热实验研究

在保护液亚临界相变温度范围进行肾脏器官的低温延时保存,是一个不同于传统低温保存的新方法。为保持细胞活性必须从器官摘取的第一步展开研究,用保护液对离体肾脏进行冷灌注是移植器官保存的重要环节,研究冷灌注过程中器官温度随时间变化的传热特征,探求提供与传统生物保存方法所不同的传热传质条件非常重要。用高倍率红外热象仪、热电偶探针、超声血流计等,对猪整体肾脏冷灌注过程进行了实时测试,得出动脉血管作为冷源时的肾脏温度动态变化曲线。实验结果表明,在1℃冷却液匀速灌注过程中肾脏温度的降低表现为非均匀变化,究其原因是由于肾脏内特定的组织结构引起。实验研究将对肾脏器官的三维温度场数值计算提供宝贵的实验依据。     

Monitoring of the influence of long‐term oxidative stress and ischemia on the condition of kidneys using solid‐phase microextraction chemical biopsy coupled with liquid chromatography–high‐resolution mass spectrometry

The limiting factor in conventional quality assessments of transplanted organs, namely the invasiveness of tissue sample collection, has prompted much research on the field of transplantology to focus on the development of alternative evaluation methods of organ quality. In the present project, we undertake the challenge to address the need for a new analytical solution for graft quality assessments by using a novel metabolomic diagnostic protocol based on low‐invasive solid‐phase microextraction. Solid‐phase microextraction probes of ca. 0.2 mm coated with 4 mm long mixed‐mode extraction phase were inserted into rabbit kidneys immediately following euthanasia and after 2, 4, 6, and 21 h of preservation. Liquid chromatography–mass spectrometry analysis of the extracts was performed with the use of a reversed phase column and a Q‐Exactive Focus mass spectrometer operated in positive ionization mode. Statistical analysis of significantly changing compounds revealed metabolic profile changes in kidneys induced by ischemia and oxidative stress as a function of the duration of cold storage. The most pronounced alterations were reflected in levels of essential amino acids and purine nucleosides. Our findings demonstrate that the proposed approach may be successfully used to monitor changes in the metabolic profile of organs over time of preservation.     

超高效液相色谱-串联质谱法检测猪肉和猪肾中残留的10种镇静剂类药物

建立了猪肉和猪肾中安眠酮、氯丙嗪、异丙嗪、地西泮、硝西泮、奥沙西泮、替马西泮、咪达唑仑、三唑仑和唑吡旦10种镇静剂类药物残留检测的超高效液相色谱-串联质谱分析方法。样品经酶解后,调节pH值呈碱性,分别用乙酸乙酯和叔丁基甲醚进行萃取,高速冷冻离心去除脂肪等杂质。提取液经Waters Acquity UPLC BEH C18色谱柱分离,以0.1%甲酸乙腈溶液和0.1%甲酸水溶液为流动相进行梯度洗脱。电喷雾正离子(ESI+)模式电离,多反应监测(MRM)模式检测,基质匹配标准溶液法进行定量。结果表明: 10种镇静剂在2～100 μg/L范围内呈良好的线性关系,相关系数(r)均大于0.998; 10种镇静剂的检出限(S/N＞3)为0.5 μg/kg,定量限(S/N＞10)为1 μg/kg。3个添加水平的回收率为64.5%～111.4%,批内、批间相对标准偏差(RSD)均小于15%。该方法具有简便快捷、灵敏度高、定性准确等特点。     

Characterization of monumental carbonate stones by thermal analysis (TG, DTG and DSC)

Thermogravimetry (TG), derivative thermogravimetry (DTG) and differential scanning calorimetry (DSC), were found to be suitable instrumental techniques for the study of monumental rocks because they need small amounts of sample and provide extensive qualitative and quantitative information. From DTG curves, the calcite/dolomite ratio in the samples as well as the differences between limestones and dolomites can be quantitatively determined. DSC curves are adequate for the identification of the degradation products in the monumental stones since they are usually salts (sometimes hydrates) which present first-order processes under 600°C. This technique makes it possible to carry out quantitative and semiquantitative analysis of the degradation of monumental rocks since it provides data about its mineralogic composition.