Proteomics reveals potential non-neuronal cholinergic receptor-effectors in endothelial cells

Aim:

The non-neuronal acetylcholine system (NNAS) in endothelial cells participates in modulating endothelial function, vascular tone, angiogenesis and inflammation, thus plays a critical role in cardiovascular diseases. In this study, we used a proteomic approach to study potential downstream receptor-effectors of NNAS that were involved in regulating cellular function in endothelial cells.

Methods:

Human umbilical vein endothelial cells were incubated in the presence of acetylcholine, oxotremorine, pilocarpine or nicotine at the concentration of 10 μmol/L for 12 h, and the expressed proteins in the cells were separated and identified with two-dimensional electrophoresis (2-DE) and LC-MS. The protein spots with the largest changes were identified by LC-MS. Biowork software was used for database search of the peptide mass fingerprints.

Results:

Over 1200 polypeptides were reproducibly detected in 2-DE with a pH range of 3–10. Acetylcholine, oxotremorine, pilocarpine and nicotine treatment caused 16, 9, 8 and 9 protein spots, respectively, expressed differentially. Four protein spots were identified as destrin, FK506 binding protein 1A (FKBP1A), macrophage migration inhibitory factor (MIF) and profilin-1. Western blotting analyses showed that treatment of the cells with cholinergic agonists significantly decreased the expression of destrin, FKBP1A and MIF, and increased the expression of profilin-1.

Conclusion:

A set of proteins differentially expressed in endothelial cells in response to cholinergic agonists may have important implications for the downstream biological effects of NNAS.     

Novel technique for the treatment of large subcapsular renal hematoma: combined use of percutaneous drainage and urokinase injection

Purpose

To evaluate the efficacy and safety of combined use of percutaneous drainage and urokinase injection to treat patients with large subcapsular renal hematoma.

Methods

We retrospectively analyzed the data of 20 patients with large subcapsular renal hematoma who were treated with percutaneous drainage and urokinase at our institutes between 2002 and 2010. Percutaneous drainage of the hematoma was performed after bleeding stopped. Then, urokinase was injected into the hematoma cavity per our protocol every 3 days. During the treatment and follow-up period, the diameter of hematoma was assessed by ultrasonography and enhanced computed tomography (CT) in all cases. Complete blood count, serum creatinine and blood pressure were evaluated as well. Follow-up periods ranged from 12 to 60 months (mean 24.1 ± 11.5 months).

Results

Percutaneous renal hematoma drainage combined with urokinase injection was successfully performed in all the patients. After sufficient drainage, all of the patients were free from their symptoms. The average time to pain relief was 10.8 ± 2.1 days after percutaneous drainage, and the total duration of drainage was 28 ± 5.1 days. Relative to baseline, there was reduction in the diameter of hematoma evaluated by the enhanced CT scan after treatment (from 11.7 ± 3.1 to 2.8 ± 1.0 cm). During long-term follow-up, active bleeding, hypertension, renal function impairment or infectious complications were not found.

Conclusions

This study revealed that combined use of percutaneous drainage and urokinase injection was a safe and effective treatment for patient with large subcapsular renal hematoma.     

Isolating and characterizing three alternatively spliced mu opioid receptor variants: mMOR‐1A,mMOR‐1O,and mMOR‐1P

Extensive alternative pre‐mRNA splicing of the mu opioid receptor gene, OPRM1, has demonstrated an array of splice variants in mice, rats and humans. Three classes of splice variants have been identified: full‐length seven transmembrane (TM) domain variants with C‐terminal splicing, truncated 6TM variants and single TM variants. The current studies isolates and characterizes an additional three full‐length C‐terminal splice variants generated from the mouse OPRM1 gene: mMOR‐1A, mMOR‐1O, and mMOR‐1P. Using RT‐qPCR, we demonstrated differential expression of these variants' mRNAs among selected brain regions, supporting region‐specific alternative splicing. When expressed in Chinese Hamster Ovary cells, all the variants displayed high mu binding affinity and selectivity with subtle differences in the affinities toward some agonists. [³⁵S]γGTP binding assays revealed marked differences in agonist‐induced G protein activation in both potency and efficacy among the variants. Together with the previous studies of mu agonist‐induced phosphorylation and internalization in several carboxyl terminal splice variants, the current studies further suggest the existence of biased signaling of various agonists within each individual variant and/or among different variants. Synapse 68:144–152, 2014 . © 2013 Wiley Periodicals, Inc.     

Is lead chelation therapy effective for chronic kidney disease? A meta‐analysis

The heavy metal lead (Pb) is a major environmental and occupational hazard. Epidemiological studies have demonstrated a strong association between lead exposure and the presence of chronic kidney injury. Some studies have suggested that chelation therapy with calcium disodium ethylenediaminetetraacetic acid (calcium disodium EDTA) might help decrease the progression of chronic kidney disease among patients with measurable body lead burdens. However, calcium disodium EDTA chelation in lead exposure is controversial due to the potential for adverse effects such as acute tubular necrosis. Therefore, we investigated the available randomized controlled trials assessing the renoprotective effects of calcium disodium EDTA chelation therapy. Our meta‐analysis shows that calcium disodium EDTA chelation therapy can effectively delay the progression of chronic kidney disease in patients with measurable body lead burdens reflected by increasing the levels of estimated glomerular filtration rate (eGFR) and creatinine clearance rate (Ccr). There appears to be no conclusive evidence that calcium disodium EDTA can decrease proteinuria.     

直肠癌术后吻合口漏的诊断与治疗进展

随着全直肠系膜切除技术的推广,中低位直肠癌的保肛手术比例大大提高.保肛术后吻合口漏是直肠癌术后最重要的并发症,也是术后发生其他并发症和死亡的主要原因.国内外的外科医师对直肠癌术后吻合口漏的早期诊断、预防及治疗进行了大量的研究,对该并发症有了较深入的认识.     

胃肠间质瘤分子分型的临床实践价值

随着分子靶向治疗在胃肠间质瘤（GIST）领域取得的巨大成功,GIST中的分子事件引起了广泛关注。研究发现．GIST患者的药效反应和预后与其本身的基因改变,即分子分型密切相关。借助分子分型,将肿瘤生物学行为相似的患者归为同一组别,进而制定相应的治疗策略,可能会成为今后GIST临床实践的重要模式。     

肛提肌外腹会阴联合直肠切除术治疗低位直肠癌

传统的腹会阴联合直肠切除术存在环周切缘阳性率高及术中穿孔率较高而影响预后的缺点,肛提肌外腹会阴联合直肠切除术可克服这些缺点．但该术式存在操作复杂、创伤大和并发症多等缺点。通过近几年的临床研究．国内外学者对该术式的适应证、手术方式改良及并发症的防治有了更加深入的认识。本文就经胍提肌外腹会阴联合直肠切除术的特点、手术适应证、研究进展及相关并发症的防治进行讨论。     

直肠癌保肛术后肠道功能障碍研究进展

直肠癌手术理念和技术的进步,使得低位直肠癌保肛率明显提高,而肠道功能障碍是困扰保肛术后患者的主要问题。目前研究从解剖、生理及动力学角度分析排粪功能改变的机制,认为肠道功能障碍与术后解剖改变、神经损伤以及括约肌功能受损等密切相关。本文总结了直肠癌术后肠道功能障碍的流行病学和发生机制,并阐述术后肠道功能障碍发生机制对预防和治疗术后肠道功能障碍的重要意义。