Vascular units as advanced living materials for bottom-up engineering of perfusable 3D microvascular networks

The timely establishment of functional neo-vasculature is pivotal for successful tissue development and regen-eration,remaining a central challenge in tissue engineering.In this study,we present a novel(micro)vascular-ization strategy that explores the use of specialized“vascular units”(VUs)as building blocks to initiate blood vessel formation and create perfusable,stroma-embedded 3D microvascular networks from the bottom-up.We demonstrate that VUs composed of endothelial progenitor cells and organ-specific fibroblasts exhibit high angiogenic potential when embedded in fibrin hydrogels.This leads to the formation of VUs-derived capillaries,which fuse with adjacent capillaries to form stable microvascular beds within a supportive,extracellular matrix-rich fibroblastic microenvironment.Using a custom-designed biomimetic fibrin-based vessel-on-chip(VoC),we show that VUs-derived capillaries can inosculate with endothelialized microfluidic channels in the VoC and become perfused.Moreover,VUs can establish capillary bridges between channels,extending the microvascular network throughout the entire device.When VUs and intestinal organoids(IOs)are combined within the VoC,the VUs-derived capillaries and the intestinal fibroblasts progressively reach and envelop the IOs.This promotes the formation of a supportive vascularized stroma around multiple IOs in a single device.These findings un-derscore the remarkable potential of VUs as building blocks for engineering microvascular networks,with ver-satile applications spanning from regenerative medicine to advanced in vitro models.

The 5-HT2c receptor gene Cys23Ser polymorphism influences the intravaginal ejaculation latency time in Dutch Caucasian men with lifelong premature ejaculation

It has been postulated that the persistent short intravaginal ejaculation latency time （IELT） of men with lifelong premature ejaculation （LPE） is related to 5-hydroxytryptamine （HT）2c receptor functioning. The aim of this study was to investigate the relationship of Cys23Ser 5-HT2c receptor gene polymorphism and the duration of IELT in men with LPE. Therefore, a prospective study was conducted in 64 Dutch Caucasian men with LPE. Baseline IELT during coitus was assessed by stopwatch over a 1-month period. All men were genotyped for Cys23Ser 5-HT2c receptor gene polymorphism. Allele frequencies and genotypes of Cys and Ser variants of 5-HT2c receptor gene polymorphism were determined. Association between Cys/Cys and Ser/Ser genotypes and the natural logarithm of the IELT in men with LPE were.investigated. As a result, the geometric mean, median and natural mean IELT were 25.2, 27.0, 33.9s, respectively. Of all men, 20.0%, 10.8%, 23.1% and 41.5% ejaculated within 10, 10-20, 20-30 and 30-60s after vaginal penetration. Of the 64 men, the Cys/Cys and Ser/Ser genotype frequency for the Cys23Ser polymorphism of the 5-HT2c receptor gene was 81% and 19%, respectively. The geometric mean IELT of the wildtypes （Cys/Cys） is significantly lower （22.6s; 95% CI 18.3-27.8s） than in male homozygous mutants （Ser/Ser） （40.4s; 95% CI 20.3-80.4s） （P = 0.03）. It is concluded that Cys23Ser 5-HT2c receptor gene polymorphism is associated with the IELT in men with LPE. Men with Cys/Cys genotype have shorter IELTs than men with Ser/Ser genotypes.

Potential therapeutic benefits stemming from the thermal nature of irreversible electropora tion of solid cancers

To the Editor：Irreversible electroporation （IRE） is a CE- and FDA- approved treatment modality for pancreatic and liver tumors that is based on the site-confined destruction of tumor tissue by multiple short, high-intensity electrical pulses.

以吡嗪酰胺用量作为评估结核病漏报的方法

目的:为了建立确认荷兰医生对活动性结核病报告的方法。 方法:以服用化疗药物吡嗪酰胺作为结核病发生的标志。根据发给门诊病人吡嗪酰胺的每日剂量(DDD)来估计1994-1998年期间荷兰结核病患者的数量。DDD是一个测量的技术单位,不必须反映推荐或者实际应用的剂量。通常它的主要指标是以具有正常器官功能成人的平均每日剂量为基础。荷兰医疗保险部(CVZ)的药物信息规划部门(GIP)提供DDD资料。以通报给荷兰结核病登记处(NTR)的结核病患者为基础,按照这些病人的体重计算他们应该服用多少吡嗪酰胺(测量DDD)。 结果:按照GIP药房记录的DDD数量与通告给NTR计算出的DDD的数量只相差8％;登记的病人数应该是6889,而不是6349。 结论:GIP和NTR测量的吡嗪酰胺用量的非常接近,有力地证明荷兰报告的结核病与应报告的疾病指标相符。这种方法使用简便并且值得在其他国家推广。

TREM-1 multimerization is essential for its activation on monocytes and neutrophils

The triggering receptor expressed on myeloid cells-1(TREM-1)is a receptor expressed on innate immune cells.By promoting the amplification of inflammatory signals that are initially triggered by Toll-like receptors(TLRs),TREM-1 has been characterized as a major player in the pathophysiology of acute and chronic inflammatory diseases,such as septic shock,myocardial infarction,atherosclerosis,and inflammatory bowel diseases.However,the molecular events leading to the activation of TREM-1 in innate immune cells remain unknown.Here,we show that TREM-1 is activated by multimerization and that the levels of intracellular Ca 2+release,reactive oxygen species,and cytokine production correlate with the degree of TREM-1 aggregation.TREM-1 activation on primary human monocytes by LPS required a two-step process consisting of upregulation followed by clustering of TREM-1 at the cell surface,in contrast to primary human neutrophils,where LPS induced a rapid cell membrane reorganization of TREM-1,which confirmed that TREM-1 is regulated differently in primary human neutrophils and monocytes.In addition,we show that the ectodomain of TREM-1 is able to homooligomerize in a concentration-dependent manner,which suggests that the clustering of TREM-1 on the membrane promotes its oligomerization.We further show that the adapter protein DAP12 stabilizes TREM-1 surface expression and multimerization.TREM-1 multimerization at the cell surface is also mediated by its endogenous ligand,a conclusion supported by the ability of the TREM-1 inhibitor LR12 to limit TREM-1 multimerization.These results provide evidence for ligand-induced,receptor-mediated dimerization of TREM-1.Collectively,our findings uncover the mechanisms necessary for TREM-1 activation in monocytes and neutrophils.

Carboxylesterase 1 family knockout alters drug disposition and lipid metabolism

The mammalian carboxylesterase 1(Ces1/CES1)family comprises several enzymes that hydrolyze many xenobiotic chemicals and endogenous lipids.To investigate the pharmacological and physiological roles of Ces1/CES1,we generated Ces1 cluster knockout(Ces1^(-/-))mice,and a hepatic human CES1 transgenic model in the Ces1^(-/-)background(TgCES1).Ces1^(-/-)mice displayed profoundly decreased conversion of the anticancer prodrug irinotecan to SN-38 in plasma and tissues.TgCES1 mice exhibited enhanced metabolism of irinotecan to SN-38 in liver and kidney.Ces1 and hCES1 activity increased irinotecan toxicity,likely by enhancing the formation of pharmacodynamically active SN-38.Ces1^(-/-)mice also showed markedly increased capecitabine plasma exposure,which was moderately decreased in TgCES1 mice.Ces1^(-/-)mice were overweight with increased adipose tissue,white adipose tissue inflammation(in males),a higher lipid load in brown adipose tissue,and impaired blood glucose tolerance(in males).These phenotypes were mostly reversed in TgCES1 mice.TgCES1 mice displayed increased triglyceride secretion from liver to plasma,together with higher triglyceride levels in the male liver.These results indicate that the carboxylesterase 1 family plays essential roles in drug and lipid metabolism and detoxification.Ces1^(-/-)and TgCES1 mice will provide excellent tools for further study of the in vivo functions of Ces1/CES1 enzymes.

Identification of common and distinct origins of human serum and breastmilk IgA1 by mass spectrometry-based clonal profiling

The most abundant immunoglobulin present in the human body is IgA. It has the highest concentrations at the mucosal lining and in biofluids such as milk and is the second most abundant class of antibodies in serum. We assessed the structural diversity and clonal repertoire of IgA1-containing molecular assemblies longitudinally in human serum and milk from three donors using a mass spectrometry-based approach. IgA-containing molecules purified from serum or milk were assessed by the release and subsequent analysis of their Fab fragments. Our data revealed that serum IgA1 consists of two distinct structural populations, namely monomeric IgA1 (∼80%) and dimeric joining (J-) chain coupled IgA1 (∼20%). Also, we confirmed that IgA1 in milk is present solely as secretory (S)IgA, consisting of two (∼50%), three (∼33%) or four (∼17%) IgA1 molecules assembled with a J-chain and secretory component (SC). Interestingly, the serum and milk IgA1-Fab repertoires were distinct between monomeric, and J-chain coupled dimeric IgA1. The serum dimeric J-chain coupled IgA1 repertoire contained several abundant clones also observed in the milk IgA1 repertoire. The latter repertoire had little to no overlap with the serum monomeric IgA1 repertoire. This suggests that human IgA1s have (at least) two distinct origins;one of these produces dimeric J-chain coupled IgA1 molecules, shared in human serum and milk, and another produces monomeric IgA1 ending up exclusively in serum.

Neuronal toll-like receptors and neuro-immunity in Parkinson’s disease,Alzheimer’s disease and stroke

Toll-like receptors(TLRs)are part of the innate immune system and can initiate an immune response upon exposure to harmful microorganisms.Neuronal TLRs are considered to be part of an established framework of interactions between the immune system and the nervous system,the major sensing systems in mammals.TLRs in the nervous system and neuronal TLRs are suspected to be important during inflammation and neurodegenerative diseases.The aim of this review is to offer an overview of the current knowledge about TLRs in neurodegenerative pathologies,with a focus on Parkinson’s disease.More research focusing on the role of TLRs in health and disease of the nervous system is needed and remains to be explored.

临床试验中的非特异性复合终点评价的危害

复合终点这一方法看似能够增加统计学效能，但是却能掩盖真正的疗效。 根据国际指南，临床试验中终点的评价应该说明治疗的风险与获益，应该与患者具有相关性，常用，且使临床试验更切实可行。为了达到这一目标，研究者们选择的终点主要包括全因病死率、所有住院率或任何不良事件。由于这些终点将多个特因死亡的终点综合在一起，所以把他们叫做复合终点。全因病死率是一种非常流行的终点评价方法，