High-throughput identification of off-targets for the mechanistic study of severe adverse drug reactions induced by analgesics

Drugs may induce adverse drug reactions (ADRs) when they unexpectedly bind to proteins other than their therapeutic targets. Identification of these undesired protein binding partners, called off-targets, can facilitate toxicity assessment in the early stages of drug development. In this study, a computational framework was introduced for the exploration of idiosyncratic mechanisms underlying analgesic-induced severe adverse drug reactions (SADRs). The putative analgesic-target interactions were predicted by performing reverse docking of analgesics or their active metabolites against human/mammal protein structures in a high-throughput manner. Subsequently, bioinformatics analyses were undertaken to identify ADR-associated proteins (ADRAPs) and pathways. Using the pathways and ADRAPs that this analysis identified, the mechanisms of SADRs such as cardiac disorders were explored. For instance, 53 putative ADRAPs and 24 pathways were linked with cardiac disorders, of which 10 ADRAPs were confirmed by previous experiments. Moreover, it was inferred that pathways such as base excision repair, glycolysis/glyconeogenesis, ErbB signaling, calcium signaling, and phosphatidyl inositol signaling likely play pivotal roles in drug-induced cardiac disorders. In conclusion, our framework offers an opportunity to globally understand SADRs at the molecular level, which has been difficult to realize through experiments. It also provides some valuable clues for drug repurposing.     

Unusual hyperpigmented patches,an undeveloped breast and a cataract in a female with incontinentia pigmenti

A 19‐year‐old female and her mother visited our department with a history of unusual reticular brown‐black patches on their trunks and limbs since infancy. Besides the obviously fulsome hyperpigmentation, the daughter also showed other unusual clinical manifestations such as generalized hypohidrosis, especially on the hyperpigmented patches, cicatricial alopecia, a cataract on her right eye, abnormal teeth and a right dysplastic breast. The mother had a median diastema between her maxillary central incisors, hypoplasia of the enamel, hypohidrosis and hyperpigmented patches on her left thoracic region. Analysis of the NEMO (NF‐κB essential modulator) gene in the patient and her mother revealed a deletion of exons 4–10. Their EDA and EDAR genes were normal.     

脑梗死并发低钠血症与营养相关因素的分析

目的:分析脑梗死病人低钠血症的发生情况以及与营养相关因素的关系,旨在提高临床综合治疗效果。方法:对240例脑梗死病人,于入院后第2天清晨空腹抽取静脉血,检测血清钠、血红蛋白、总蛋白、清蛋白、C-反应蛋白和淋巴细胞计数,并进行统计学分析。结果:240例脑梗死病人发生低钠血症者为61例,占25.42%。低钠组病人血清总蛋白、清蛋白、血红蛋白、淋巴细胞计数均明显低于血钠正常组,C-反应蛋白显著高于血钠正常组(P<0.05)。脑梗死病人血清钠与总蛋白、清蛋白、血红蛋白和C-反应蛋白之间具有明显相关性(P<0.05)。结论:脑梗死病人低钠血症与机体营养不良因素密切相关,在纠正低钠血症的同时,要加强病人营养支持,从而提高临床综合治疗的效果。     

The Value of Routinely Performed Ultrasonography in Patients with Crohn Disease

Background: In active Crohn disease (CD), abdominal ultrasound can demonstrate morphological changes in the bowel wall. By contrast, the role of ultrasonography in periods with no disease activity has never been evaluated. In this prospective study we investigated the outcome of routinely performed abdominal ultrasonography in CD patients irrespective of symptoms and disease activity. Methods: 255 consecutive patients with chronic CD (117 M, 138 F, mean age ( ± s ) 38 ± 14) were evaluated by high-resolution ultrasonography of the bowel wall and abdomen. The findings were graded with respect to further diagnostic and/or therapeutic implications. In addition, the CD activity index (CDAI) was determined. In patients with pathological sonographic findings, complementary procedures (e.g. endoscopy, computed tomography, biopsy or operation) were performed to validate the suspected diagnosis. Results: In 46/255 (18%) patients with CD, the diagnosis of a transmural inflammatory reaction (TMR) with or without fistula was made by ultrasonography. In respect of CDAI, 29/46 (63%) of these patients were graded as active disease (CDAI >150) and 17/46 (37%) as inactive disease (CDAI &#104 150). Of the 17 patients with a TMR and CDAI &#104 150, 4 patients revealed interenteric fistula, 7 patients mesenteric or perirectal fistula, whereas 6 patients presented with a transmural mesenteric inflammatory reaction without fistula. All fistulae were confirmed by radiography. Ultrasonography of the remaining abdominal organs revealed pathological findings with further diagnostic implications in 25/255 (10%) patients and with therapeutic implications in 9/255 (4%) patients. Conclusion: Routinely performed ultrasonography of the abdomen reveals pathological findings with therapeutic implications not only in symptomatic but also in asymptomatic patients with CD. It can therefore be recommended as a screening tool for this group of patients.     

C反应蛋白在鉴别系统性红斑狼疮活动与合并感染中的意义

目的通过分析系统性红斑狼疮(system ic lupus erythem atosus,SLE)病情活动和合并感染患者的临床和血清学特点,发现对疾病活动和感染有鉴别意义的指标。方法对2003—2004年在我科住院的110例SLE患者进行回顾性分析,比较病情活动未合并感染者87例(未感染组)和合并感染者23例(感染组)的发热、皮疹、口腔溃疡、脏器损害等临床表现,同时分析患者的C反应蛋白(CRP),血沉(ESR),血清蛋白电泳(白蛋白、α1球蛋白、α2球蛋白、β球蛋白、γ球蛋白),补体(C3、C4),免疫球蛋白(IgA、IgM、IgG)及免疫复合物(CIC)的变化。结果感染组CRP、ESR及α1球蛋白高于未感染组(P<0.01)。通过比较CRP、ESR及α1球蛋白与其他临床指标的相关性,发现CR P受感染之外的影响较少,在未感染组CRP浓度的均数在正常水平,合并感染后明显升高。感染组患者的发热、口腔溃疡、雷诺现象、心脏损害及糖皮质激素用量均高于未感染组(P<0.05)。结论譹SLE合并感染时大部分患者的CR P明显升高。譺在狼疮活动时CR P大多正常或略高,但少数患者,尤其合并浆膜炎者的CR P水平可较高。CRP对于鉴别狼疮活动与合并感染有重要的临床应用价值。     

Beneficial effect of a novel pentadecapeptide BPC 157 on gastric lesions induced by restraint stress,ethanol, indomethacin,and capsaicin neurotoxicity

Very recently, the integrity of capsaicin somatosensory neurons and their protection were suggested to be related to the activity in nociception of a newly discovered 15-amino acid peptide, BPC 157, shown to have strong beneficial effect on intestinal and liver lesions. Therefore, from this viewpoint, we have studied the gastroprotective effect of the pentadecapeptide BPC 157, on gastric lesions produced in rats by 96% ethanol, restraint stress, and indomethacin. The possible involvement of sensory neurons in the salutary actions of BPC 157 (10µg/kg, 10 ng/kg intraperitoneally) was studied with capsaicin, which has differential effects on sensory neurons: a high dose in adult (125 mg/kg subcutaneously, 3 months old) or administration (50 mg/kg subcutaneously) to neonatal animals (age of the 7 days) destroys sensory fibers, whereas a low dose (500µg/kg intraperitoneally) activates neurotransmitter release and protective effects on the mucosa. In the absence of capsaicin, BPC 157 protected gastric mucosa against ethanol, restraint, and indomethacin application. In the presence of neurotoxic doses of capsaicin, the negative influence of capsaicin on restraint, ethanol, or indomethacin lesions consistently affected salutary activity of BPC 157. However, BPC 157 protection was still evident in the capsaicin-treated rats (either treated as adults or as newborns) in all of these assays. Interestingly, after neonatal capsaicin treatment, a complete abolition of BPC gastroprotection was noted if BPC 157 was applied as a single nanogram-regimen, but the mucosal protection was fully reversed when the same dose was used daily. In line with the excitatory dose of capsaicin the beneficial effectiveness of BPC 157 appears to be increased as well. Taken together, these data provide evidence for complex synergistic interaction between the beneficial effectiveness of BPC 157 and peptidergic sensory afferent neuron activity.     

Developmental synaptic regulator,TWEAK/Fn14 signaling,is a determinant of synaptic function in models of stroke and neurodegeneration

Identifying molecular mediators of neural circuit development and/or function that contribute to circuit dysfunction when aberrantly reengaged in neurological disorders is of high importance. The role of the TWEAK/Fn14 pathway, which was recently reported to be a microglial/neuronal axis mediating synaptic refinement in experience-dependent visual development, has not been explored in synaptic function within the mature central nervous system. By combining electrophysiological and phosphoproteomic approaches, we show that TWEAK acutely dampens basal synaptic transmission and plasticity through neuronal Fn14 and impacts the phosphorylation state of pre- and postsynaptic proteins in adult mouse hippocampal slices. Importantly, this is relevant in two models featuring synaptic deficits. Blocking TWEAK/Fn14 signaling augments synaptic function in hippocampal slices from amyloid-beta–overexpressing mice. After stroke, genetic or pharmacological inhibition of TWEAK/Fn14 signaling augments basal synaptic transmission and normalizes plasticity. Our data support a glial/neuronal axis that critically modifies synaptic physiology and pathophysiology in different contexts in the mature brain and may be a therapeutic target for improving neurophysiological outcomes.

Neural circuit patterning, refinement, and plasticity are enabled by the dynamic strengthening, weakening, and pruning of chemical synapses in response to circuit activity. However, synapse loss and reduced plasticity are early hallmarks of chronic neurological disorders such as autism, schizophrenia and Alzheimer’s disease (AD) (1–3). It is therefore hypothesized that the underlying molecular mechanisms of pruning, although normally balanced in health, are dysregulated in disease. Particularly interesting is the notion that the mechanisms responsible for the reduction in functional synapses in disease reflect the aberrant reactivation of pathways important for synapse elimination in development. For example, in an AD model, synapse elimination was shown to be mediated by the complement pathway in the hippocampus (HC), reflecting aberrant reactivation of complement-dependent synapse elimination that occurs in the dorsal lateral geniculate nucleus (dLGN) of the thalamus during visual development (4). In such a paradigm, the reactivation of developmental mechanisms enables pathways that can act universally across different ages, circuits, and brain regions. Thus, the mechanisms underlying normal circuit development and their potential reactivation as key contributors to neurological diseases are areas of deep interest.In addition to chronic neurological disorders, circuitry changes also occur in acute ischemic stroke, the second leading cause of death worldwide and a cause of debilitating long-term disability. Interruptions in blood flow that deprive neurons of oxygen and nutrients result in significant cell death, followed by deficits in neurophysiological activity that are associated with poor motor recovery (5). Remarkably, the adult brain can undergo some degree of spontaneous poststroke recovery, apparently by engaging neuroplasticity mechanisms including remapping, synaptogenesis, and synaptic strengthening (5, 6). Despite these adaptations, over half of ischemic stroke patients fail to recover completely and continue to experience persistent long-term disability (7). The underlying signaling pathways that regulate synaptic physiology after stroke are an active topic of investigation.TNF-like weak inducer of apoptosis (TWEAK) protein, originally discovered as a cytokine produced by macrophages (8), signals through its injury-inducible transmembrane receptor, FGF-inducible molecule-14 (Fn14) (9). Consequently, the function of TWEAK/Fn14 signaling was elucidated as a driver of tissue remodeling in contexts of injury and disease in a variety of organ systems (10). Recently, findings have suggested a role for the TWEAK/Fn14 pathway in the central nervous system (CNS). Namely, several compelling observations indicate that TWEAK signaling through Fn14 might be a key molecular modulator of synaptic function in contexts of neurological challenge. TWEAK and Fn14 are up-regulated in the CNS in AD (11, 12, 13 and SI Appendix, Fig. S6A) and after ischemic stroke in humans and mice (14–16). Importantly, TWEAK/Fn14 signaling was also recently shown to be a pathway necessary for synapse maturation during experience-dependent visual development. Light-induced up-regulation of Fn14 in thalamocortical excitatory neurons and corresponding up-regulation of TWEAK in microglia mediate the elimination of weak synapses and strengthening of remaining synapses in the dLGN (17, 18). Indeed, the communication between neurons and supporting microglia has emerged as a key mechanism regulating neuronal circuitry, with microglia deploying their ramified processes to continuously survey and refine synapses in response to neural activity. Interestingly, TWEAK expression has also been shown to be microglia-enriched in the mouse cortex (19), suggesting that it may play a role in multiple brain regions. Thus, like the complement pathway, the TWEAK/Fn14 pathway could be an important regulator of synapse biology in visual development which is re-engaged and acts generally in different ages and brain regions to contribute to pathology.The involvement of TWEAK/Fn14 signaling in synapse physiology or pathophysiology outside of the developing visual system is unknown. We considered it to be a strong candidate modifier of synaptic function in adults given that Fn14 is up-regulated and required for synaptic refinement in experience-dependent visual development, and TWEAK and Fn14 are up-regulated in contexts of neurological injury/disease, suggesting that the TWEAK/Fn14 system is tuned to periods of substantial change in neuronal activity levels or environment (e.g., eye opening, ischemic stroke). We employed HC slices to test the hypothesis that the TWEAK/Fn14 pathway regulates synaptic function in adult mice and in different disease contexts and delineate its mechanism of action. Herein, we reveal that TWEAK, through neuronal Fn14, mediates acute dampening of basal synaptic transmission and synaptic plasticity in hippocampal slices from mature mice. Furthermore, we demonstrate that TWEAK/Fn14 signaling broadly impacts the phosphorylation state of critical synaptic proteins, suggesting a general role in synapse modulation. Finally, we show that pathway deficiency or pharmacological inhibition of TWEAK/Fn14 signaling augments synaptic transmission and plasticity in amyloid-beta (Aβ)–overexpressing mice and post ischemic stroke animals, two model systems featuring synaptic functional deficits. Thus, our results support that TWEAK/Fn14 constitutes a synaptic regulatory pathway with therapeutic potential for CNS disorders in the adult brain.     

CD40 在癫痫大鼠大脑皮层及海马中的表达变化

目的：研究CD40炎症分子在癫痫持续状态(status epilepticus,SE)后大鼠大脑皮层及海马的表达。方法：采 用免疫组织化学及免疫荧光双标记方法,观察锂-匹罗卡品癫痫大鼠大脑皮层及海马的不同区域、不同时间点、不同 细胞中CD40的表达情况。结果：癫痫发作显著增加了CD40阳性细胞,以在海马组织中增加为著; SE后CD40主要在 激活的小胶质细胞上表达;CD40阳性细胞在SE后3 d增加达到一个高峰,在SE 7 d后恢复到SE前稍高的水平。结论： CD40在激活的小胶质细胞上高表达,促进了SE大鼠海马炎症反应,提示CD40介导的信号通路参与SE后海马组织的炎 症病理过程。