The role of sequestosome 1/p62 protein in amyotrophic lateral sclerosis and frontotemporal dementia pathogenesis

Amyotrophic lateral sclerosis and frontotemporal lobar degeneration are multifaceted diseases with genotypic,pathological and clinical overlap.One such overlap is the presence of SQSTM1/p62 mutations.While traditionally mutations manifesting in the ubiquitin-associated domain of p62 were associated with Paget’s disease of bone,mutations affecting all functional domains of p62 have now been identified in amyotrophic lateral sclerosis and frontotemporal lobar degeneration patients.p62 is a multifunctional protein that facilitates protein degradation through autophagy and the ubiquitin-proteasome system,and also regulates cell survival via the Nrf2 antioxidant response pathway,the nuclear factor-kappa B signaling pathway and apoptosis.Dysfunction in these signaling and protein degradation pathways have been observed in amyotrophic lateral sclerosis and frontotemporal lobar degeneration,and mutations that affect the role of p62 in these pathways may contribute to disease pathogenesis.In this review we discuss the role of p62 in these pathways,the effects of p62 mutations and the effect of mutations in the p62 modulator TANK-binding kinase 1,in relation to amyotrophic lateral sclerosis-frontotemporal lobar degeneration pathogenesis.

The development and application of Laboratory Animal Science in China

The 14th Scientific Congress of the Chinese Association for Laboratory Animal Science was held on October 11‐12,2018 in Qingdao,China.During this congress,an international forum on the development of Laboratory Animal Sciences(LAS)worldwide was held in which participants learnt about the development of new LAS resources and technologies,as well as the progress of LAS in China.The main points that were discussed are as follows.After nearly a century of development,there are more than 200 species of experimental animals,and more than 20 000 inbred lines,outbred lines,genetic engineered animals,animal models of diseases,and other resources all over the world,which provide abundant experimental animal resources for scientific research.These resources are widely used in life science research.

Control of nuclear-cytoplasmic shuttling of Ankrd54 by PKCδ

AIM To identify and characterize the effect of phosphorylation on the subcellular localization of Ankrd54.METHODS HEK293 T cells were treated with calyculin A, staurosporin or phorbol 12-myristate 13-acetate(PMA). Cells were transfected with eG FP-tagged Ankrd54 with or without Lyn tyrosine kinase(wild-type, Y397 F mutant, or Y508 F mutant). The subcellular localization was assessed by immunofluorescence imaging of cells, immunoblotting of subcellular fractionations. The phosphorylation of Ankrd54 was monitored using Phos-tagT M gel retardation. Phosphorylated peptides were analysed by multiplereaction-monitoring(MRM) proteomic analysis.RESULTS Activation of PKC kinases using PMA promoted nuclear export of Ankrd54 and correlated with increased Ankrd54 phosphorylation, assayed using Phos-tag TM gel retardation. Co-expression of an active form of the PKCδisoform specifically promoted both phosphorylation and cytoplasmic localization of Ankrd54, while PKCδ, Akt and PKA did not. Alanine mutation of several serine residues in the amino-terminal region of Ankrd54(Ser14, Ser17, Ser18, Ser19) reduced both PMA induced cytoplasmic localization and phosphorylation of Ankrd54. Using MRM proteomic analysis, phosphorylation of the Ser18 residue of Ankrd54 was readily detectable in response to PMA stimulation. PMA stimulation of cells co-expressing Ankrd54 and Lyn tyrosine kinase displayed increased coimmunoprecipitation and enhanced co-localization in the cytoplasm.CONCLUSION We identify phosphorylation by PKCδ as a major regulator of nuclear-cytoplasmic shuttling of Ankrd54, and its interaction with the tyrosine kinase Lyn.

根除幽门螺杆菌，从合理使用抗生素做起

幽门螺杆菌根除失败的主要原因是对抗生素产生耐药。由于目前我国对治疗幽门螺杆菌的认识不统一、治疗不规范,随着抗生素的广泛使用,导致幽门螺杆菌对多种抗生素耐药率呈上升趋势,根除率下降。为了提高幽门螺杆菌的根除率,减少抗生素耐药,本文对治疗幽门螺杆菌的七种抗生素的药理机制和幽门螺杆菌对抗生素的耐药机制进行了概述,从而加深临床医生对治疗幽门螺杆菌的抗生素的认识,促使其治疗规范化。同时呼吁临床医生在治疗前进行胃黏膜细菌培养和药敏试验,从而科学规范用药,减少耐药菌株的出现,缩短治疗进程,避免资源浪费,提高幽门螺杆菌的根除率,降低消化性溃疡的复发率,延缓胃黏膜相关淋巴组织(MALT)淋巴瘤的进程,预防胃癌的产生,造福患者。

Multipotent pancreas progenitors: Inconclusive but pivotal topic

The establishment of multipotent pancreas progenitors(MPP) should have a significant impact not only on the ontology of the pancreas, but also for the translational research of glucose-responding endocrine b-cells. Deficiency of the latter may lead to the pandemic type 1 or type 2 diabetes mellitus, a metabolic disorder. An ideal treatment of which would potentially be the replacement of destroyed or failed b-cells, by restoring function of endogenous pancreatic endocrine cells or by transplantation of donor islets or in vitro generated insulin-secreting cells. Thus, considerable research efforts have been devoted to identify MPP candidates in the preand post-natal pancreas for the endogenous neogenesis or regeneration of endocrine insulin-secreting cells. In order to advance this inconclusive but critical field, we here review the emerging concepts, recent literature and newest developments of potential MPP and propose measures that would assist its forward progression.

Insulin-secreting β cells require a post-genomic concept

Pancreatic insulin-secretingβcells are essential in maintaining normal glucose homeostasis accomplished byhighly specialized transcription of insulin gene,of which occupies up to 40%their transcriptome.Deficiency of these cells causes diabetes mellitus,a global public health problem.Although tremendous endeavors have been made to generate insulin-secreting cells from human pluripotent stem cells(i.e.,primitive cells capable of giving rise to all cell types in the body),a regenerative therapy to diabetes has not yet been established.Furthermore,the nomenclature ofβcells has become inconsistent,confusing and controversial due to the lack of standardized positive controls of developmental stagematched in vivo cells.In order to minimize this negative impact and facilitate critical research in this field,a postgenomic concept of pancreaticβcells might be helpful.In this review article,we will briefly describe howβcells were discovered and islet lineage is developed that may help understand the cause of nomenclatural controversy,suggest a post-genomic definition and finally provide a conclusive remark on future research of this pivotal cell.

Strategies for generating mouse model resources of human disease

The number of genetically modified mouse models that mimic human disease is growing rapidly,but only a tiny fraction has been commonly used.According to The Knockout Mouse Program(Lloyd,2011),a public resource of mouse embryonic stem cells containing a null mutation in every gene in the mouse genome,8,916 mutant mice lines were phenotyped up to 19 July 2022.Due to the poor correlation between the genomic responses in the mouse models and those responses in human disease,and since humans differ significantly in their genetic vulnerability to common diseases,we still need better mouse models,especially for common and chronic human diseases,including cancer,pulmonary and cardiovascular diseases,obesity and diabetes,behavioral disorders,and neurodegenerative diseases.These new models will be placed into a public repository,The China National Human Disease Animal Model Resource Center(NAMR).This project is funded by Ministry of Science and Technology of China and specializes in the creation,introduction,collection,preservation,and supply of animal model resources forhuman diseases.

The genetic reference population “Collaborative Cross”is a powerful resource for genetic discoveries and understanding complex genetic traits

The Collaborative Cross( CC) is a powerful genetic resource with a wide range of applications in medical research. It is a multiparental genetic reference population,composed of 200 recombinant inbred mouse strains. It was designed to be a resource for rapid mapping of genes that mediate complex traits. The CC has been produced following an intense breeding program that had taken over ten years to complete. It is now becoming available to the research community. While advances in human genetic technologies over the last decade have been substantial,the CC provides the ability to make discoveries not possible with other methods or resources. It can be applied to any subject that can be measured or modelled in the mouse. Here,we discuss a range of applications for which the CC can be used,with examples taken from the early characterization of this powerful resource.

Novel hybrid biocomposites for tendon grafts:The addition of silk to polydioxanone and poly(lactide-co-caprolactone)enhances material properties,in vitro and in vivo biocompatibility

Biopolymers play a critical role as scaffolds used in tendon and ligament(TL)regeneration.Although advanced biopolymer materials have been proposed with optimised mechanical properties,biocompatibility,degradation,and processability,it is still challenging to find the right balance between these properties.Here,we aim to develop novel hybrid biocomposites based on poly(p-dioxanone)(PDO),poly(lactide-co-caprolactone)(LCL)and silk to produce high-performance grafts suitable for TL tissue repair.Biocomposites containing 1-15%of silk were studied through a range of characterisation techniques.We then explored biocompatibility through in vitro and in vivo studies using a mouse model.We found that adding up to 5%silk increases the tensile properties,degradation rate and miscibility between PDO and LCL phases without agglomeration of silk inside the composites.Furthermore,addition of silk increases surface roughness and hydrophilicity.In vitro experiments show that the silk improved attachment of tendon-derived stem cells and proliferation over 72 h,while in vivo studies indicate that the silk can reduce the expression of pro-inflammatory cytokines after six weeks of implantation.Finally,we selected a promising biocomposite and created a prototype TL graft based on extruded fibres.We found that the tensile properties of both individual fibres and braided grafts could be suitable for anterior cruciate ligament(ACL)repair applications.

An objective measure for the assessment and management of fluid shifts in acute major burns

Background: Major burns are life threatening. Fluid resuscitation is required for survival to maintain intravascular volumes and prevent hypovolemic shock. Bioimpedance spectroscopy (BIS) has been recognised as a potential method of monitoring fluid shifts after burn and in other disease states. The aims of this study were to examine the reliability of BIS across different dressing conditions and electrode positions, establish the influence of Acticoat?on BIS variable measures and determine the validity of whole-body BIS to assess net fluid shift in the presence of moderate to major burns. Methods: An observational longitudinal cohort study was conducted from December 2014 to February 2016. Patients with over 15% total body surface area (TBSA) burns and injury less than 48 h were enrolled in the study. BIS triplicate measures were collected in an open wound and with an ActicoatTMdressing (at 5 half hour intervals). Standard and alternate electrode placements were utilised for the reliability analysis and standard placement only for determining the validity of BIS in moderate to major burns. The ImpediMde SFB7 was used to collect whole-body and segmental BIS measures. Stata statistical software, release 14 was utilised to analyse all results. Descriptive analyses were performed and were reported using the means and standard deviations (SD). Results: BIS-repeated measures established BIS raw resistance (R), and predicted volume variables were reliable in any condition (intra-class correlation coefficient (ICC) 0.996-0.999, 95% confidence intervals (CI) 0.996-0.999) without a systematic difference. Acticoat?dressings significantly influenced all BIS-predicted volumes (p≤0.01) as determined by multilevel mixed effects (MLME) linear regression analysis. Validity of BIS was demonstrated by resistance variables significantly decreasing with increasing net ionic fluid shift and increased TBSA (severity of injury) and calculated fluid volumes increasing with increasing net fluid shift and TBSA. BIS resistance also decreased with time as oedema reduced. For clinical use, a calculator was developed to adjust BIS variables when an Acticoat?dressing is in situ, thus facilitating BIS variable change estimates in real time, with dressings intact. Conclusion: BIS may be used clinically to monitor fluid volume change in major acute burns.

Demonstration of the test-retest reliability and sensitivity of the Lower Limb Functional Index-10 as a measure of functional recovery post burn injury:a cross-sectional repeated measures study design

Background:Lower limb burns can significantly delay recovery of function.Measuring lower limb functional outcomes is challenging in the unique burn patient population and necessitates the use of reliable and valid tools.The aims of this study were to examine the test-retest reliability,sensitivity,and internal consistency of Sections 1 and 3 of the Lower Limb Functional Index-10(LLFI-10)questionnaire for measuring functional ability in patients with lower limb burns over time.Methods:Twenty-nine adult patients who had sustained a lower limb burn injury in the previous 12 months completed the test-retest procedure of the study.In addition,the minimal detectable change(MDC)was calculated for Section 1 and 3 of the LLFI-10.Section 1 is focused on the activity limitations experienced by patients with a lower limb disorder whereas Section 3 involves patients indicating their current percentage of pre-injury duties.Results:Section 1 of the LLFI-10 demonstrated excellent test-retest reliability(intra-class correlation coefficient(ICC)0.98,95%CI 0.96–0.99)whilst Section 3 demonstrated high test-retest reliability(ICC 0.88,95%CI 0.79–0.94).MDC scores for Sections 1 and 3 were 1.27 points and 30.22%,respectively.Internal consistency was demonstrated with a significant negative association(rs=?0.83)between Sections 1 and 3 of the LLFI-10(p<0.001).Conclusions:This study demonstrates that Section 1 and 3 of the LLFI-10 are reliable for measuring functional ability in patients who have sustained lower limb burns in the previous 12 months,and furthermore,Section 1 is sensitive to changes in patient function over time.

Natural,synthetic and commercially-available biopolymers used to regenerate tendons and ligaments

Tendon and ligament(TL)injuries affect millions of people annually.Biopolymers play a significant role in TL tissue repair,whether the treatment relies on tissue engineering strategies or using artificial tendon grafts.The biopolymer governs the mechanical properties,biocompatibility,degradation,and fabrication method of the TL scaffold.Many natural,synthetic and hybrid biopolymers have been studied in TL regeneration,often combined with therapeutic agents and minerals to engineer novel scaffold systems.However,most of the advanced biopolymers have not advanced to clinical use yet.Here,we aim to review recent biopolymers and discuss their features for TL tissue engineering.After introducing the properties of the native tissue,we discuss different types of natural,synthetic and hybrid biopolymers used in TL tissue engineering.Then,we review biopolymers used in commercial absorbable and non-absorbable TL grafts.Finally,we explain the challenges and future directions for the development of novel biopolymers in TL regenerative treatment.

Pharmacologic RNA splicing modulation: a novel mechanism to enhance neoantigen-directed anti-tumor immunity and immunotherapy response

A recent study by Lu et al.1 suggests a novel approach to increasing responsiveness to immune checkpoint blockade (ICB) therapy. ICB is a promising form of cancer immunotherapy that aims to boost the anti-tumoral immune response. This response is primarily driven by the presentation of different types of antigens at the cancer cell membrane via the type I major histocompatibility complex (MHC I). Neoantigens are tumor-specific antigens that are small (mostly 9-mers) mutated peptides that generally result from somatic mutations. High tumor mutational burden, which results in the formation of many neoantigens, is currently one of the main biomarkers for ICB response prediction. Because ICB is only effective in a minority of patients, new therapeutic strategies are required to augment this response.

Cancer susceptibility genes: update and systematic perspectives

A cancer susceptibility gene(CSG)is a gene whose variants can increase the cancer risk of those bearing such variants.1 By defining an individual’s CSG profile,it may be possible to assess their cancer risk and may provide insights for future prevention and treatment of cancer.The methods used for discovery of CSGs have evolved from single candidate gene analysis to population-based next-generation sequencing.