Large animal models for Huntington's disease research

Huntington'sdisease(HD)isahereditary neurodegenerative disorder for which there is currently no effectivetreatmentavailable.Consequently,the development of appropriate disease models is critical to thoroughly investigate disease progression.The genetic basis of HD involves the abnormal expansion of CAG repeats in the huntingtin(HTT)gene,leading to the expansion of a polyglutamine repeat in the HTT protein.Mutant HTT carrying the expanded polyglutamine repeat undergoes misfolding and forms aggregates in the brain,which precipitate selective neuronal loss in specific brain regions.Animal models play an important role in elucidating the pathogenesis of neurodegenerative disorders such as HD and in identifying potential therapeutic targets.Due to the marked species differences between rodents and larger animals,substantial efforts have been directed toward establishing large animal models for HD research.These models are pivotal for advancing the discovery of novel therapeutic targets,enhancing effective drug delivery methods,and improving treatment outcomes.We have explored the advantages of utilizing large animal models,particularly pigs,in previous reviews.Since then,however,significant progress has been made in developing more sophisticated animal models that faithfully replicate the typical pathology of HD.In the current review,we provide a comprehensive overview of large animal models of HD,incorporating recent findings regarding the establishment of HD knock-in(KI)pigs and their genetic therapy.We also explore the utilization of large animal models in HD research,with a focus on sheep,non-human primates(NHPs),and pigs.Our objective is to provide valuable insights into the application of these large animal models for the investigation and treatment of neurodegenerative disorders.

长链非编码RNA HOXA远端转录本在口腔鳞状细胞癌中的表达及生物学意义

目的:探讨长链非编码RNA HOXA远端转录本(HOXA transcript at the distal tip,HOTTIP)在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)中的表达及对OSCC细胞生物学功能的影响。方法:收集2020年1月至2021年6月在郑州大学第一附属医院接受手术治疗的52例OSCC患者,采用RT-qPCR检测HOTTIP在OSCC组织和癌旁组织以及人OSCC细胞株HSC-4、SCC-9和CAL-27和人正常口腔角质细胞株HOK中的表达水平,并分析其与OSCC患者临床病理资料及总生存期(overall survival,OS)的关系。采用CCK-8、流式细胞术、Transwell和Western blot分析下调HOTTIP或共下调HOTTIP和miR-637对CAL-27细胞增殖、凋亡、迁移、侵袭和上皮间质转化(epithelial-mesenchymal transition,EMT)能力的影响。采用双荧光素酶报告基因实验分析HOTTIP和miR-637的靶向关系。结果:与癌旁组织相比,OSCC组织中HOTTIP的表达水平明显升高(P<0.05),且与肿瘤大小、TNM分期、分化程度、淋巴结转移及OS相关(P<0.05)。与人正常口腔角质HOK细胞相比,HSC-4、SCC-9和CAL-27细胞中HOTTIP的表达明显升高(P<0.05)。下调HOTTIP后,CAL-27细胞的增殖、迁移、侵袭及EMT能力明显减弱,细胞凋亡率明显升高(P<0.05)。HOTTIP与miR-637具有靶向关系,且下调HOTTIP后,CAL-27细胞中miR-637表达明显升高(P<0.05)。同时下调HOTTIP和miR-637后,CAL-27细胞的增殖、迁移、侵袭与EMT能力明显增强,细胞凋亡率明显降低(P<0.05)。结论:HOTTIP在OSCC组织和细胞中高表达,可靶向miR-637调控OSCC细胞的增殖、凋亡、迁移、侵袭和EMT能力。

Current applications and future prospects of nanotechnology in cancer immunotherapy

Cancer immunotherapy is an artificial stimulation of the immune system to recognize cancer cells and activate specific immune cells to target and attack cancer cells.In clinical trials, immunotherapy has recently shown impressive results in the treatment of multiple cancers.Thus, cancer immunotherapy has gained a lot of attention for its unique advantages and promising future.With extensive research on cancer immunotherapy, its safety and effectiveness has gradually been revealed.However, it is still a huge challenge to expand and drive this therapy while maintaining low toxicity, high specificity, and long-lasting efficacy.As a unique technology, nanotechnology has been applied in many fields, the advantages of which will promote the development of cancer immunotherapies.Researchers have tried to apply nanomaterials to cancer immunotherapy due to their advantageous properties,such as large specific surface areas, effective drug delivery, and controlled surface chemistry, to improve treatment efficacy.Here,we briefly introduce the current applications of nanomaterials in cancer immunotherapy, including adoptive cell therapy(ACT),therapeutic cancer vaccines, and monoclonal antibodies, and throw light on future directions of nanotechnology-based cancer immunotherapy.

Differential development and electrophysiological activity in cultured cortical neurons from the mouse and cynomolgus monkey

In vitro cultures of primary cortical neurons are widely used to investigate neuronal function.However,it has yet to be fully investigated whether there are significant differences in development and function between cultured rodent and primate cortical neurons,and whether these differences influence the utilization of cultured cortical neurons to model pathological conditions.Using in vitro culture techniques combined with immunofluorescence and electrophysiological methods,our study found that the development and maturation of primary cerebral cortical neurons from cynomolgus monkeys were slower than those from mice.We used a microelectrode array technique to compare the electrophysiological differences in cortical neurons,and found that primary cortical neurons from the mouse brain began to show electrical activity earlier than those from the cynomolgus monkey.Although cultured monkey cortical neurons developed slowly in vitro,they exhibited typical pathological features-revealed by immunofluorescent staining-when infected with adeno-associated viral vectors expressing mutant huntingtin(HTT),the Huntington’s disease protein.A quantitative analysis of the cultured monkey cortical neurons also confirmed that mutant HTT significantly reduced the length of neurites.Therefore,compared with the primary cortical neurons of mice,cultured monkey cortical neurons have longer developmental and survival times and greater sustained physiological activity,such as electrophysiological activity.Our findings also suggest that primary cynomolgus monkey neurons cultured in vitro can simulate a cell model of human neurodegenerative disease,and may be useful for investigating time-dependent neuronal death as well as treatment via neuronal regeneration.All mouse experiments and protocols were approved by the Animal Care and Use Committee of Jinan University of China(IACUC Approval No.20200512-04)on May 12,2020.All monkey experiments were approved by the IACUC protocol(IACUC Approval No.LDACU 20190820-01)on August 23,2019 for animal management and use.

Sonic Hedgehog stimulates migration of MCF-7 breast cancer cells through Rac1

As one of the most common tumors in women, breast cancer has drawn considerable interest from investigators and clinicians in recent years. Despite early diagnosis and best therapeutic regimens available, the prognosis of malignant or metastatic breast cancer patients is still not optimistic. Hedgehog signaling, a classical pathway indispensable to embryonic development, participates in the growth of a variety of tumors. In the present study,the effect of Sonic Hedgehog(Shh) on breast cancer cells was investigated. We identified that Shh signal stimulated the migration of MCF-7 breast cancer cells. Smo and Gli1 were involved in Shh-stimulated migration of MCF-7 cells. Activating Smo and Gli1 induced cell migration, which was blocked by their specific antagonists.The effect of Shh signaling on MCF-7 cells was independent of Wnt5 a, Dvl2 and Rab35, but directly dependent on Rac1. In conclusion, our study suggested that Shh promotes breast cancer cell migration via Rac1 independently of the non-canonical Wnt signaling pathway, which may represent a rational molecular target for combination medication in breast cancer.

Extracellular magnetic labeling of biomimetic hydrogel-induced human mesenchymal stem cell spheroids with ferumoxytol for MRI tracking

Labeling of mesenchymal stem cells(MSCs)with superparamagnetic iron oxide nanoparticles(SPIONs)has emerged as a potential method for magnetic resonance imaging(MRI)tracking of transplanted cells in tissue repair studies and clinical trials.Labeling of MSCs using clinically approved SPIONs(ferumoxytol)requires the use of transfection reagents or magnetic field,which largely limits their clinical application.To overcome this obstacle,we established a novel and highly effective method for magnetic labeling of MSC spheroids using ferumoxytol.Unlike conventional methods,ferumoxytol labeling was done in the formation of a mechanically tunable biomimetic hydrogel-induced MSC spheroids.Moreover,the labeled MSC spheroids exhibited strong MRI T2 signals and good biosafety.Strikingly,the encapsulated ferumoxytol was localized in the extracellular matrix(ECM)of the spheroids instead of the cytoplasm,minimizing the cytotoxicity of ferumoxytol and maintaining the viability and stemness properties of biomimetic hydrogel-induced MSC spheroids.This demonstrates the potential of this method for post-transplantation MRI tracking in the clinic.

A MOF/poly(thioctic acid)composite for enhanced gold extraction from water matrices

With the fast generation of electronic waste(e-waste)and the increasing depletion of metal resources,“urban mining”that can selectively recover gold from secondary resources has attracted great interest.Construction of materials with high extraction capacity and satisfying selectivity in complex aqueous-based matrices still remains challenging.Here,a novel metal-organic framework/polymer composite(Fe-BTC/poly(thioctic acid),denoted as Fe-BTC/pTA)has been newly synthesized and applied for selective gold recovery in different matrices(river water,seawater,and leaching solution of e-waste).Benefiting from the high specific surface area and suitable pore sizes as well as the rational design of active sites,the composite exhibits high adsorption capacity(920 mg/g),high removal efficiency(>99%),fast kinetics(below 0.1 ppb within 10 min),and good applicability in complex matrices,which are better than those of most reported sulfur-containing adsorbents.Solid-state metallic gold with high purity can be effectively enriched due to the high recyclability and long-term stability of the composite.The material after adsorption can be further applied as a heterogeneous catalyst for water remediation due to the in situ generated gold nanoparticles by the redox reaction between Au(III)ions and the S-containing groups in the composites.

Pre-mating nitenpyram exposure in male mice leads to depression-like behavior in offspring by affecting tryptophan metabolism in gut microbiota

Several studies have confirmed that the health status of the paternal affects the health of the offspring,however,it remains unknown whether paternal exposure to pesticides affect the offspring health.Here,we used untargeted metabolomics and 16S rRNA sequencing technology,combined with tail suspension test and RT-qPCR to explore the effects of paternal exposure to nitenpyram on the neurotoxicity of offspring.Our results found that the paternal exposure to nitenpyram led to the offspring’s depressive-like behaviors,accompanied by the reduction of tryptophan content and the disorder of microbial abundance in the gut of the offspring.Further,we determined the expression of tryptophan metabolism-related genes tryptophanase(tnaA)and tryptophan hydroxylase 1(TpH1)in gut bacteria and colonic tissues.We found that tryptophan is metabolized to indoles rather than being absorbed into colonocytes,which coursed the reduce of tryptophan availability after nitenpyram exposure.In conclusion,our study deepens our understanding of the intergenerational toxic effects of pesticides.

Optical responses of metallic plasmonic arrays under the localized excitation

The metallic plasmonic array that can support both propagating surface plasmon polaritons(PSPPs)and localized surface plasmon resonance(LSPR)possesses rich optical properties and remarkable optical performance,making it a powerful platform for applications in photonics,chemistry,and materials.For practical applications,the excitation spot is usually smaller than the area of metal arrays.It is thus imperative to address“how many array units are enough?”towards a rational design of plasmonic nanostructures.Herein,we employed focused ion beam(FIB)to precisely fabricate a series of plasmonic array structures with increased unit number.By utilizing photoluminescence(PL)and surface-enhanced Raman spectroscopy(SERS),we found that the array units outside the excitation spot still have a significant impact on the optical response within the spot.Combined with the numerical simulation,we found that the boundary of the finite array leads to the loss of PSPP outside the excitation point,which subsequently affects the coupling of PSPP and LSPR in the excitation spot,leading to variations in PL and SERS intensity.Based on the findings,we further tuned the LSPR mode of the metal arrays by electrodeposition to obtain strong near-field enhancement without any influence on the PSPP mode.This work advances the understanding of near-field and far-field optical behavior in finite-size array structures and provides guidance for designing highly-efficient photonic devices.

PINK1 kinase dysfunction triggers neurodegeneration in the primate brain without impacting mitochondrial homeostasis

In vitro studies have established the prevalent theory that the mitochondrial kinase PINK1 protects neurodegeneration by removing damaged mitochondria in Parkinson's disease(PD).However,difficulty in detecting endogenous PINK1 protein in rodent brains and cell lines has prevented the rigorous investigation of the in vivo role of PINK1.Here we report that PINK1 kinase form is selectively expressed in the human and monkey brains.CRISPR/Cas9-mediated deficiency of PINK1 causes similar neurodegeneration in the brains of fetal and adult monkeys as well as cultured monkey neurons without affecting mitochondrial protein expression and morphology.Importantly,PINK1 mutations in the primate brain and human cells reduce protein phosphorylation that is important for neuronal function and survival.Our findings suggest that PINK1 kinase activity rather than its mitochondrial function is essential for the neuronal survival in the primate brains and that its kinase dysfunction could be involved in the pathogenesis of PD.

Novel magnetic silk fibroin scaffolds with delayed degradation for potential long-distance vascular repair

Although with the good biological properties,silk fibroin(SF)is immensely restrained in long-distance vascular defect repair due to its relatively fast degradation and inferior mechanical properties.It is necessary to construct a multifunctional composite scaffold based on SF.In this study,a novel magnetic SF scaffold(MSFCs)was prepared by an improved infiltration method.Compared with SF scaffold(SFC),MSFCs were found to have better crystallinity,magnetocaloric properties,and mechanical strength,which was ascribed to the rational introduction of iron-based magnetic nanoparticles(MNPs).Moreover,in vivo and in vitro experiments demonstrated that the degradation of MSFCs was significantly extended.The mechanism of delayed degradation was correlated with the dual effect that was the newly formed hydrogen bonds between SFC and MNPs and the complexing to tyrosine(Try)to inhibit hydrolase by internal iron atoms.Besides,theβ-crystallization of protein in MSFCs was increased with the rise of iron concentration,proving the beneficial effect after MNPS doped.Furthermore,although macrophages could phagocytose the released MNPs,it did not affect their function,and even a reasonable level might cause some cytokines to be upregulated.Finally,in vitro and in vivo studies demonstrated that MSFCs showed excellent biocompatibility and the growth promotion effect on CD34-labeled vascular endothelial cells(VECs).In conclusion,we confirm that the doping of MNPs can significantly reduce the degradation of SFC and thus provide an innovative perspective of multifunctional biocomposites for tissue engineering.

Maintaining the balance of TDP-43,mitochondria,and autophagy:a promising therapeutic strategy for neurodegenerative diseases

Mitochondria are the energy center of cell operations and are involved in physiological functions and maintenance of metabolic balance and homeostasis in the body.Alterations of mitochondrial function are associated with a variety of degenerative and acute diseases.As mitochondria age in cells,they gradually become inefficient and potentially toxic Acute injury can trigger the permeability of mitochondrial membranes,which can lead to apoptosis or necrosis.Transactive response DNA-binding protein 43 kDa(TDP-43)is a protein widely present in cells.It can bind to RNA,regulate a variety of RNA processes,and play a role in the formation of multi-protein/RNA complexes.Thus,the normal physiological functions of TDP-43 are particularly important for cell survival.Normal TDP-43 is located in various subcellular structures including mitochondria,mitochondrial-associated membrane,RNA particles and stress granules to regulate the endoplasmic reticulum-mitochondrial binding,mitochondrial protein translation,and mRNA transport and translation.Importantly,TDP-43 is associated with a variety of neurodegenerative diseases,including amyotrophic lateral sclerosis,frontotemporal dementia and Alzheimer's disease,which are characterized by abnormal phosphorylation,ubiquitination,lysis or nuclear depletion of TDP-43 in neurons and glial cells.Although the pathogenesis of TDP-43 proteinopathy remains unknown,the presence of pathological TDP-43 inside or outside of mitochondria and the functional involvement of TDP-43 in the regulation of mitochondrial morphology,transport,and function suggest that mitochondria are associated with TDP-43-related diseases.Autophagy is a basic physiological process that maintains the homeostasis of cells,including targeted clearance of abnormally aggregated proteins and damaged organelles in the cytoplasm;therefore,it is considered protective against neurodegenerative diseases.However,the combination of abnormal TDP-43 aggregation,mitochondrial dysfunction,and insufficient autophagy can lead to a variety of aging-related pathologies.In this review,we describe the current knowledge on the associations of mitochondria with TDP-43 and the role of autophagy in the clearance of abnormally aggregated TDP-43 and dysfunctional mitochondria.Finally,we discuss a novel approach for neurodegenerative treatment based on the knowledge.

Continuous synthesis of extremely small-sized iron oxide nanoparticles used for T_(1)-weighted magnetic resonance imaging via a fluidic reactor

Extremely small-sized iron oxide nanoparticles(ESIONPs)with sizes less than 5 nm have shown great promise as T_(1)contrast agents for magnetic resonance imaging(MRI).However,their facile and scalable production with simultaneously endowed biocompatible surface chemistry remains difficult to be realized.In this study,by using the coprecipitation method implemented in a specially designed gas/liquid mixed phase fluidic reactor,polyglucose sorbitol carboxymethyether(PSC)coated ESIONPs were continuously synthesized with controllable particle sizes ranging from 1.8 to 4 nm.Among the differently sized ESIONPs,the 3.7-nm ESIONPs exhibit the best performance as T_(1)MRI contrast agent,featuring a high r_(1) value of 4.11(mmol L^(−1))^(−1)s^(−1)and low r_(2)/r_(1) ratio of 7.90 under a clinical 3 T MR scanning,as well as the excellent T_(1)MRI contrast effect in not only water but also the cellular environment and blood vessel.Furthermore,the ESIONPs possess long-term stability and good dispersity in aqueous dispersions,making them ideal candidates as safe and effective T_(1)-weighted MRI contrast agent for real clinical use.

Experimental study on the acoustic propagation and anisotropy of coal rocks

In order to study the propagation laws of acoustic wave of coal samples from the Upper Permian Xuanwei Formation in the east of Yunnan Province,China,under saturated water and dry conditions,the basic physical parameters,acoustic parameters and anisotropic parameters were obtained through the experiments.Based on FFT and wavelet analysis theory,the spectral characteristics of coal samples under different conditions were studied.The results show that physical parameters of coal samples in different directions have different values,that is,the anisotropy of coal samples is obvious.When the coal samples are saturated with water,the acoustic velocities and the attenuation coefficient increase,whereas the dominant frequency decreases.The signal amplitude of the frequency domain significantly decreases,that is,the internal structure of coal samples is damaged.The P-wave velocity and S-wave velocity increase with the increase of the confining pressure,whereas the anisotropy parameters decrease with the increase of the confining pressure.Overall,this study provides the basis to understand basic acoustic information and anisotropy characteristics of coal samples.

Ceramic-based dielectric metamaterials

Dielectric metamaterials based on ceramics have attracted considerable in-terest in the past few years owing to their low dielectric loss,simple structure,excellent multifield tunability,and good environmental adaptability.They are considered to be promising alternative to metal-based metamaterials and can lead to a new strategy for the development of passive devices.In this review,the recent progress of ceramic-based dielectric metamaterials in electro-magnetic applications,energy applications,non-Hermitian systems,and natural materials with near-zero or negative refraction are summarized.The design principle and mechanism,as well as manufacturing technologies,are also introduced,and the current development trend of ceramic-based dielectric metamaterials are proposed.

Flexible organic integrated electronics for self-powered multiplexed ocular monitoring

Smart contact lens has drawn extensive research interests due to the noninvasive real-time detection of the human body to provide biomedical information for health management.However,it has been difficult to accurately measure the physiological signals in tears,and the use of external power source has also hindered the future applications.Here,we demonstrated an organic electrochemical transistor based multiplexed sensors self-powered by the organic solar cells(OSCs).The integrated device was fabricated via simple process including solution blade-coating and thermal evaporation.OSCs were optimized to provide optimal operation voltage for the sensors that exhibit semilog-linear response to the glucose and calcium ions in tear fluids without any peripheral circuits.The sensing signals can be transmitted to the laptop wirelessly through a near filed communication unit.This integrated self-powered multiplexed sensing device will provide real-time monitoring of the biomarkers in tears,prospected to be installed on the smart contact lens for the early detection and diagnosis of diabetes.

Discovery of a highly specific ^(18)F-labeled PET ligand for phosphodiesterase 10A enabled by novel spirocyclic iodonium ylide radiofluorination

As a member of cyclic nucleotide phosphodiesterase(PDE)enzyme family,PDE10A is in charge of the degradation of cyclic adenosine(cAMP)and guanosine monophosphates(cGMP).While PDE10A is primarily expressed in the medium spiny neurons of the striatum,it has been implicated in a variety of neurological disorders.Indeed,inhibition of PDE10A has proven to be of potential use for the treatment of central nervous system(CNS)pathologies caused by dysfunction of the basal ganglia–of which the striatum constitutes the largest component.A PDE10A-targeted positron emission tomography(PET)radioligand would enable a better assessment of the pathophysiologic role of PDE10A,as well as confirm the relationship between target occupancy and administrated dose of a given drug candidate,thus accelerating the development of effective PDE10A inhibitors.In this study,we designed and synthesized a novel ^(18)F-aryl PDE10A PET radioligand,codenamed[^(18)F]P10A-1910([^(18)F]9),in high radiochemical yield and molar activity via spirocyclic iodonium ylide-mediated radiofluorination.[^(18)F]9 possessed good in vitro binding affinity(IC_(50)=2.1 nmol/L)and selectivity towards PDE10A.Further,[^(18)F]9 exhibited reasonable lipophilicity(logD=3.50)and brain permeability(P_(app)>10×10^(−6) cm/s in MDCK-MDR1 cells).PET imaging studies of[^(18)F]9 revealed high striatal uptake and excellent in vivo specificity with reversible tracer kinetics.Preclinical studies in rodents revealed an improved plasma and brain stability of[^(18)F]9 when compared to the current reference standard for PDE10A-targeted PET,[^(18)F]MNI659.Further,dose–response experiments with a series of escalating doses of PDE10A inhibitor 1 in rhesus monkey brains confirmed the utility of[^(18)F]9 for evaluating target occupancy in vivo in higher species.In conclusion,our results indicated that[^(18)F]9 is a promising PDE10A PET radioligand for clinical translation.

Generation of inactivated IL2RG and RAG1 monkeys with severe combined immunodeficiency using base editing

Severe combined immunodeficiency(SCiD)encompasses a range of inherited disorders that lead to a profound deterioration of the immune system.Among the pivotal genes associated with SCID,RAG1 and IL2RG play crucial roles.IL2RG is essential for the development,differentiation,and functioning of T,B,and NK cells,while RAG1 critically contributes to adaptive immunity by facilitating V(D)J recombination during the maturation of lymphocytes.Animal models carrying mutations in these genes exhibit notable deficiencies in their immune systems.

Tauopathy promotes spinal cord-dependent production of toxic amyloid-beta in transgenic monkeys

Tauopathy,characterized by the hyperphosphorylation and accumulation of the microtubule-associated protein tau,and the accumulation of Aβ oligomers,constitute the major pathological hallmarks of Alzheimer's disease.However,the relationship and causal roles of these two pathological changes in neurodegeneration remain to be defined,even though they occur together or independently in several neurodegenerative diseases associated with cognitive and movement impairment.