A DNA tetrahedron-based ferroptosis-suppressing nanoparticle: superior delivery of curcumin and alleviation of diabetic osteoporosis

Diabetic osteoporosis(DOP)is a significant complication that poses continuous threat to the bone health of patients with diabetes;however,currently,there are no effective treatment strategies.In patients with diabetes,the increased levels of ferroptosis affect the osteogenic commitment and differentiation of bone mesenchymal stem cells(BMSCs),leading to significant skeletal changes.To address this issue,we aimed to target ferroptosis and propose a novel therapeutic approach for the treatment of DOP.We synthesized ferroptosis-suppressing nanoparticles,which could deliver curcumin,a natural compound,to the bone marrow using tetrahedral framework nucleic acid(tFNA).This delivery system demonstrated excellent curcumin bioavailability and stability,as well as synergistic properties with tFNA.Both in vitro and in vivo experiments revealed that nanoparticles could enhance mitochondrial function by activating the nuclear factor E2-related factor 2(NRF2)/glutathione peroxidase 4(GPX4)pathway,inhibiting ferroptosis,promoting the osteogenic differentiation of BMSCs in the diabetic microenvironment,reducing trabecular loss,and increasing bone formation.These findings suggest that curcumin-containing DNA tetrahedron-based ferroptosissuppressing nanoparticles have a promising potential for the treatment of DOP and other ferroptosis-related diseases.

Comprehensive analysis of the gut microbiome and posttranslational modifications elucidates the route involved in microbiota-host interactions

The gut microbiome interacts with the host to maintain body homeostasis,with gut microbial dysbiosis implicated in many diseases.However,the underlying mechanisms of gut microbe regulation of host behavior and brain functions remain unclear.This study aimed to elucidate the influence of gut microbiota on brain functions via post-translational modification mechanisms in the presence or absence of bacteria without any stimulation.We conducted succinylome analysis of hippocampal proteins in germ-free(GF)and specific pathogen-free(SPF)mice and metagenomic analysis of feces from SPF mice.These results were integrated with previously reported hippocampal acetylome and phosphorylome data from the same batch of mice.Subsequent bioinformatics analyses revealed 584 succinylation sites on 455 proteins,including 54 up-regulated succinylation sites on 91 proteins and 99 down-regulated sites on 51 proteins in the GF mice compared to the SPF mice.We constructed a panoramic map of gut microbiota-regulated succinylation,acetylation,and phosphorylation,and identified cross-talk and relative independence between the different types of post-translational modifications in modulating complicated intracellular pathways.Pearson correlation analysis indicated that 13 taxa,predominantly belonging to the Bacteroidetes phylum,were correlated with the biological functions of post-translational modifications.Positive correlations between these taxa and succinylation and negative correlations between these taxa and acetylation were identified in the modulation of intracellular pathways.This study highlights the hippocampal physiological changes induced by the absence of gut microbiota,and proteomic quantification of succinylation,phosphorylation,and acetylation,contributing to our understanding of the role of the gut microbiome in brain function and behavioral phenotypes.

Expert consensus on irrigation and intracanal medication in root canal therapy

Chemical cleaning and disinfection are crucial steps for eliminating infection in root canal treatment. However, irrigant selection or irrigation procedures are far from clear. The vapor lock effect in the apical region has yet to be solved, impeding irrigation efficacy and resulting in residual infections and compromised treatment outcomes.

Expert consensus on difficulty assessment of endodontic therapy

Endodontic diseases are a kind of chronic infectious oral disease. Common endodontic treatment concepts are based on the removal of inflamed or necrotic pulp tissue and the replacement by gutta-percha. However, it is very essential for endodontic treatment to debride the root canal system and prevent the root canal system from bacterial reinfection after root canal therapy(RCT). Recent research, encompassing bacterial etiology and advanced imaging techniques, contributes to our understanding of the root canal system’s anatomy intricacies and the technique sensitivity of RCT. Success in RCT hinges on factors like patients, infection severity, root canal anatomy, and treatment techniques. Therefore, improving disease management is a key issue to combat endodontic diseases and cure periapical lesions. The clinical difficulty assessment system of RCT is established based on patient conditions, tooth conditions, root canal configuration, and root canal needing retreatment, and emphasizes pre-treatment risk assessment for optimal outcomes. The findings suggest that the presence of risk factors may correlate with the challenge of achieving the high standard required for RCT. These insights contribute not only to improve education but also aid practitioners in treatment planning and referral decision-making within the field of endodontics.

Experts consensus on the procedure of dental operative microscope in endodontics and operative dentistry

The dental operative microscope has been widely employed in the field of dentistry,particularly in endodontics and operative dentistry,resulting in significant advancements in the effectiveness of root canal therapy,endodontic surgery,and dental restoration.However,the improper use of this microscope continues to be common in clinical settings,primarily due to operators’insufficient understanding and proficiency in both the features and established operating procedures of this equipment.In October 2019,Professor Jingping Liang,Vice Chairman of the Society of Cariology and Endodontology,Chinese Stomatological Association,organized a consensus meeting with Chinese experts in endodontics and operative dentistry.The objective of this meeting was to establish a standard operation procedure for the dental operative microscope.Subsequently,a consensus was reached and officially issued.Over the span of about four years,the content of this consensus has been further developed and improved through practical experience.

Molecular events in the jaw vascular unit:A traditional review of the mechanisms involved in inflammatory jaw bone diseases

Inflammatory jaw bone diseases are common in stomatology,including periodontitis,peri-implantitis,medication-related osteonecrosis of the jaw,radiation osteomyelitis of the jaw,age-related osteoporosis,and other specific infections.These diseases may lead to tooth loss and maxillofacial deformities,severely affecting patients'quality of life.Over the years,the reconstruction of jaw bone deficiency caused by inflammatory diseases has emerged as a medical and socioeconomic challenge.Therefore,exploring the pathogenesis of inflammatory diseases associated with jaw bones is crucial for improving prognosis and developing new targeted therapies.Accumulating evidence indicates that the integrated bone formation and dysfunction arise from complex interactions among a network of multiple cell types,including osteoblast-associated cells,immune cells,blood vessels,and lymphatic vessels.However,the role of these different cells in the inflammatory process and the'rules'with which they interact are still not fully understood.Although many investigations have focused on specific pathological processes and molecular events in inflammatory jaw diseases,few articles offer a perspective of integration.Here,we review the changes and mechanisms of various cell types in inflammatory jaw diseases,with the hope of providing insights to drive future research in this field.

With two episodes of right retromandibular angle subcutaneous emphysema during right upper molar crown preparation:A case report

BACKGROUND Subcutaneous emphysema is a well-known complication of oral surgery,especially during mandibular wisdom tooth extraction.However,subcutaneous emphysema secondary to dental procedures such as crown preparation is rare.The main symptom of emphysema is swelling and crepitus on palpation.Uncontrolled emphysema may spread along the fascial planes and cause deep space infections or a pneumomediastinum.CASE SUMMARY In this paper,we report a 34-year-old female who underwent upper molar tooth preparation for crowns and subsequently developed extensive subcutaneous emphysema on the retromandibular angle on two different occasions.The treatment plan for this patient involved close observation of the airway,and administration of dexamethasone and antibiotics via intravenous drip or orally.Ice bag compression was quickly applied and medication was prescribed to alleviate discomfort and promote healing.Although the main reason is unclear,the presence of a fissure in the molar is an important clue which may contribute to the development of subcutaneous emphysema during crown preparation.It is imperative for dental professionals to recognize such pre-disposing factors in order to minimize the risk of complications.CONCLUSION This case highlights the need for prompt diagnosis and management of subcutaneous emphysema because of the risk of much more serious complications.Awareness of relatively“benign”subcutaneous emphysema during any dental procedure is critical not only for inexperienced dentists,but also for those who work in rural and remote settings as members of surgical teams.In this study,we review the clinical presentation,mechanism,and differential diagnosis of subcutaneous emphysema.

Clinicopathological study of malignant peripheral nerve sheath tumors in the head and neck:Case reports and review of literature

BACKGROUND Malignant peripheral nerve sheath tumor(MPNST)is a rare and aggressive soft tissue sarcoma that poses a major diagnostic and therapeutic challenge.CASE SUMMARY We retrospectively reviewed patients with head and neck MPNSTs treated in our hospital from 2000 to 2021.The clinical features,pathological manifestations,treatments,and prognoses were summarized.We also reviewed the literature,focusing on MPNST in the mandible and maxilla.The study population consisted of five women and five men aged 22–75 years(mean age,49 years).Of the 10 patients,7 were initial cases and 3 were recurrent cases.All lesions were sporadic.The most common site was the mandible.The most frequently encountered symptoms were a progressive mass and local swelling.Complete or partial loss of trimethylation at lysine 27 of histone H3(H3K27me3)was evident on staining in four of nine cases(one case was excluded due to lack of tissue for evaluation of loss of H3K27me3).The 2-and 5-year disease-specific survival rates were 86%a nd 43%,respectively.The average survival time was 64 mo.CONCLUSION MPNST is a highly malignant tumor with a poor prognosis,prone to a high risk of recurrence and distant metastasis.Complete surgical resection is the main treatment.

Identification of microbial metabolites that accelerate the ubiquitindependent degradation of c-Myc

Myc belongs to a family of proto-oncogenes that encode transcription factors.The overexpression of c-Myc causes many types of cancers.Recently,we established a system for screening c-Myc inhibitors and identified antimycin A by screening the RIKEN NPDepo chemical library.The specific mechanism of promoting tumor cell metastasis by high c-Myc expression remains to be explained.In this study,we screened approximately 5,600 microbial extracts using this system and identified a broth prepared from Streptomyces sp.RK19-A0402 strongly inhibits c-Myc transcriptional activity.After purification of the hit broth,we identified compounds closely related to the aglycone of cytovaricin and had a structure similar to that of oligomycin A.Similar to oligomycin A,the hit compounds inhibited mitochondrial complex V.The mitochondria dysfunction caused by the compounds induced the production of reactive oxygen species(ROS),and the ROS activated GSK3α/βthat phosphorylated c-Myc for ubiquitination.This study provides a successful screening strategy for identifying natural products as potential c-Myc inhibitors as potential anticancer agents.

Expert consensus on digital guided therapy for endodontic diseases

Digital guided therapy(DGT)has been advocated as a contemporary computer-aided technique for treating endodontic diseases in recent decades.The concept of DGT for endodontic diseases is categorized into static guided endodontics(SGE),necessitating a meticulously designed template,and dynamic guided endodontics(DGE),which utilizes an optical triangulation tracking system.Based on cone-beam computed tomography(CBCT)images superimposed with or without oral scan(OS)data,a virtual template is crafted through software and subsequently translated into a 3-dimensional(3D)printing for SGE,while the system guides the drilling path with a real-time navigation in DGE.DGT was reported to resolve a series of challenging endodontic cases,including teeth with pulp obliteration,teeth with anatomical abnormalities,teeth requiring retreatment,posterior teeth needing endodontic microsurgery,and tooth autotransplantation.Case reports and basic researches all demonstrate that DGT stand as a precise,time-saving,and minimally invasive approach in contrast to conventional freehand method.This expert consensus mainly introduces the case selection,general workflow,evaluation,and impact factor of DGT,which could provide an alternative working strategy in endodontic treatment.

Effect and Significance of Comprehensive Training in Infection Prevention and Control on the Psychological Health of Oral Health Staff

Background:Oral health staff have close contact with patients in the process of diagnosis and treatment,and it is inevitable for them to come into contact with patients’secretions.Therefore,oral health staff are at greater risk of infectious diseases in their daily work,and their psychological health is also greatly challenged.Objective:To study the effect and significance of comprehensive training in infection prevention and control on the psychological health of oral health staff.Methods:We selected 400 oral health staff from a tertiary stomatological hospital in Guangzhou,China in this study.The respondents were randomly divided into an intervention group and a control group.After the first round of investigation,the intervention group received comprehensive training in prevention and control of infection for three months,while the control group received no intervention.Results:The comprehensive training in infection prevention and control improved the respondents’psychological health and job satisfaction.Further strengthening infection prevention and control training for oral health staff will increase their self-confidence,improve their mental health,and increase their job satisfaction.Conclusion:For oral health staff,it is particularly important to formulate an effective and operable preventive and control training program and then implement it in a standardized manner.

Down-regulation of survivin expression by small interfering RNA induces pancreatic cancer cell apoptosis and enhances its radiosensitivity

瞄准：在导致 PC-2 房间 apoptosis 并且提高它的放射敏感度在胰腺的癌症房间线 PC-2 和 siRNA 的角色在 survivin 的表示上调查小介入 RNA (siRNA ) 的禁止的效果。方法：对 survivin 的 siRNA 原生质标志表示向量被构造并且 transfected 进有 Lipofectamine 2000 的 PC-2 房间。下面 survivin 表示的规定被半量的 RT-PCR 和免疫检测组织化学的 SP 方法和在导致 PC-2 房间 apoptosis 并且提高它的放射敏感度的 siRNA 的角色被流动血细胞计数检测。结果：顺序特定的 siRNA 高效地并且明确地下面调整在 mRNA 和蛋白质的 survivin 的表示铺平。表示抑制比率在半量的 RT-PCR 检测的 mRNA 水平是 81.25% ， 74.24% 由免疫在蛋白质水平检测了组织化学的方法。在 transfection 以后的 48 个小时， apoptosis 被 siRNA 在 7.03% 房间导致，在 14.58% ，由 siRNA 的房间与放射结合了。结论：对 survivin 的 siRNA 原生质标志表示向量能高效地并且明确地在 PC-2 房间禁止 survivin 的表示。禁止 survivin 的表示能导致 PC-2 房间的 apoptosis 并且显著地提高它的放射敏感度。对 survivin 的 RNAi 具有在胰腺的癌症的基因治疗的潜在的价值。

Science Letters:Simultaneous determination of fluoride, chloride, sulfate, phosphate, monofluorophosphate, glycerophosphate, sorbate, and saccharin in gargles by ion chromatography

Simple, reliable and sensitive analytical methods to determine anticariogenic agents, preservatives, and artificial sweeteners contained in commercial gargles are necessary for evaluating their effectiveness, safety, and quality. An ion chroma-tography (IC) method has been described to analyze simultaneously eight anions including fluoride, chloride, sulfate, phosphate, monofluorophosphate, glycerophosphate (anticariogenic agents), sorbate (a preservative), and saccharin (an artificial sweetener) in gargles. In this IC system, we applied a mobile phased gradient elution with KOH, separation by IonPac AS18 columns, and suppressed conductivity detection. Optimized analytical conditions were further evaluated for accuracy. The relative standard deviations (RSDs) of the inter-day's retention time and peak area of all species were less than 0.938% and 8.731%, respectively, while RSDs of 5-day retention time and peak area were less than 1.265% and 8.934%, respectively. The correlation coefficients for targeted analytes ranged from 0.999 7 to 1.000 0. The spiked recoveries for the anions were 90%~102.5%. We concluded that the method can be applied for comprehensive evaluation of commercial gargles.

Determination of fluorinated quinolone antibacterials by ion chromatography with fluorescence detection

For preparing fluorinated quinolone antibiotic medicine locally used in stomatology, simultaneous determination of norfloxacin, ciprofloxacin, and enoxacin was carried out by multiphase ion chromatography with fluorescence detection. Quinolone antibiotics were separated by Dionex OmniPac PAX-500 column with an eluent of 15 mmol/L H2SO4 and 35% methanol (v/v) at a flow-rate of 1.0 ml/min and detected with fluorescence with excitation and emission wave lengths of 347 nm and 420 nm respectively. The detection limits (S/N=3) of norfloxacin, ciprofloxacin and enoxacin were 50, 105 and 80 ng/ml respectively. The relative standard deviations of retention time, peak area and peak height were less than 1.1% and good linear relationship resulted. The developed method was applied to pharmaceutical formulations and biological fluids.

Characterization of oral bacterial diversity of irradiated patients by high-throughput sequencing

The objective of this study was to investigate the compositional profiles and microbial shifts of oral microbiota during head-and-neck radiotherapy.Bioinformatic analysis based on 16S rRNA gene pyrosequencing was performed to assess the diversity and variation of oral microbiota of irradiated patients.Eight patients with head and neck cancers were involved in this study.For each patient, supragingival plaque samples were collected at seven time points before and during radiotherapy.A total of 147 232 qualified sequences were obtained through pyrosequencing and bioinformatic analysis,representing 3 460 species level operational taxonomic units(OTUs) and 140 genus level taxa.Temporal variations were observed across different time points and supported by cluster analysis based on weighted UniFrac metrics.Moreover,the low evenness of oral microbial communities in relative abundance was revealed by Lorenz curves.This study contributed to a better understanding of the detailed characterization of oral bacterial diversity of irradiated patients.

An in vitro study on the efficacy of removing calcium hydroxide from curved root canal systems in root canal therapy

To compare the efficacy of various irrigants(citric acid, ethylenediaminetetraacetic acid(EDTA) and Na OCl) and techniques in removing Ca(OH)2in two types of curved root canal systems, simulated root canals with specific curvatures were used to investigate the effects of different irrigants and instruments on Ca(OH)2removal. The optimal methods were verified on extracted human teeth. Simulated root canals were assigned to one of two groups based on the irrigation solution: 10% citric acid or2.5% Na OCl. Each group was divided into four subgroups according to the technique used to remove Ca(OH)2. The percentage of Ca(OH)2removal in different sections of root canals was calculated. On the basis of the results obtained for the simulated canals, 10% citric acid and 17% EDTA were applied to remove Ca(OH)2from the extracted human teeth with curved root canal systems. The teeth were scanned by micro computed tomography to calculate the percentage of Ca(OH)2removal in the canals.In simulated root canals, we found that 10% citric acid removed more Ca(OH)2than 2.5% NaOCl in the 0–1 mm group from the apex level(Po0.05). Ultrasonic and Endo Activator activation significantly removed more Ca(OH)2than a size 30 K file in the apical third(Po0.05). However, there were no significant differences in any sections of the canals for 10% citric acid or 17%EDTA in removing Ca(OH)2in extracted human teeth. We concluded that it was effective to remove residual Ca(OH)2using the decalcifying solution with Endo Activator or Passive Ultrasonic Irrigation in a curved root canal system. A protocol for Ca(OH)2removal was provided based on the conclusions of this study and the methods recommended in previous studies.

On the road to smart biomaterials for bone research: definitions, concepts, advances, and outlook

The demand for biomaterials that promote the repair,replacement,or restoration of hard and soft tissues continues to grow as the population ages.Traditionally,smart biomaterials have been thought as those that respond to stimuli.However,the continuous evolution of the field warrants a fresh look at the concept of smartness of biomaterials.This review presents a redefinition of the term“Smart Biomaterial”and discusses recent advances in and applications of smart biomaterials for hard tissue restoration and regeneration.To clarify the use of the term“smart biomaterials”,we propose four degrees of smartness according to the level of interaction of the biomaterials with the bio-environment and the biological/cellular responses they elicit,defining these materials as inert,active,responsive,and autonomous.Then,we present an up-to-date survey of applications of smart biomaterials for hard tissues,based on the materials’responses(external and internal stimuli)and their use as immune-modulatory biomaterials.Finally,we discuss the limitations and obstacles to the translation from basic research(bench)to clinical utilization that is required for the development of clinically relevant applications of these technologies.

c-Fos expression within the L_5 spinal cord dorsal horn after spinal nerve ligation in rats Is intraplantar administration of glutamate different from intrathecal administration?

BACKGROUND： Injection of glutamate （Glu） in normal animals can cause neuronal c-Fos expression; however, whether Glu can induce spinal neuronal c-Fos expression in pain models is unclear. OBJECTIVE： To examine the effects of intraplantar and intrathecal injection of Glu on c-Fos expression in the L5 spinal cord dorsal horn Ⅰ/Ⅱ and Ⅲ/Ⅳ layers after spinal nerve ligation, and to study the effects of the N-methyI-D-aspartic acid （NMDA） receptor antagonist, D-2-amino-5-phosphonopentanoate （D-AP5）, and a selective group I mGluR antagonist, 7-hydroyiminocyclo propan[a]chromen-lacarboxylic acid ethyl ester （cpccoEt）. DESIGN, TIME AND SETTING： A randomized, controlled animal study was performed at the Department of Pharmacology, Oral Anatomy, and Neurobiology, Osaka University Graduate School of Dentistry, from December 2005 to December 2006. MATERIALS： Glu （5 μmol）, D-AP5 （50 nmot） and cpccoEt （250 nmol） were provided by Wako Pure Chemical Industries, Osaka, Japan, and diluted in saline （50 μL）. The pH of all solutions was adjusted to 7.4. METHODS： Twelve rats were randomly divided into sham operation （n = 6） and spinal nerve ligation （SNL; n = 6） groups for behavioral assessments of neuropathic pain after ligation surgery of the left L5-6 nerve segment. Another 60 rats were randomly divided into sham operation, SNL, saline-intraplantar, saline-intrathecal, Glu-intraplantar, Glu-intrathecal, D-AP5-intrathecal, Glu-D-AP5-intrathecal, cpccoEt-intrathecal, and Glu-cpccoEt-intrathecal groups, with 6 rats in each group. All groups except sham operation group received a similar SNL. On day 14, rats received a 50-μL injection of saline, Glu, D-AP5, and/or cpccoEt into the left intraplantar or intrathecal L5-4 segments. MAIN OUTCOME MEASURES： The number of c-Fos positive neurons in both Ⅰ/Ⅱ and Ⅲ/Ⅳ spinal layers at L6 was observed using immunohistochemistry 2 hours after administration. RESULTS： （1） SNL increased the level of c-Fos expression in two sides of the spinal cord, particularly on Ⅲ/Ⅳ spinal layers of the ligated side. （2） Intraplantar or intrathecal administration of saline significantly increased the c-Fos labeled neurons in Ⅰ/Ⅱ spinal layers of the ligated side, compared with SNL alone （P 〈 0.01）. （3） Intraplantar Glu （5 μmol） increased the number of c-Fos positive neurons in Ⅰ/Ⅱ spinal layers compared with intraplantar saline （P〈 0.01）. （4） The number of c-Fos neurons in Ⅰ/Ⅱ spinal layers on both the ipsilateral and contralateral side after intraplantar Glu was lower than intrathecal Glu （P〈 0.01）, with a 3-fold higher induction by intrathecal Glu. （5） Co-administration of D-AP5 or cpccoEt reduced the effects of intrathecal Glu （P 〈 0.01）. CONCLUSION： Intrathecal Glu increases c-Fos expression more than intraplantar Glu. Antagonists of NMDA and group I mGluRs block this effect.

Human β-defensin 3 gene modification promotes the osteogenic differentiation of human periodontal ligament cells and bone repair in periodontitis

Efforts to control inflammation and achieve better tissue repair in the treatment of periodontitis have been ongoing for years.Humanβ-defensin 3,a broad-spectrum antimicrobial peptide has been proven to have a variety of biological functions in periodontitis;however,relatively few reports have addressed the effects of human periodontal ligament cells(h PDLCs)on osteogenic differentiation.In this study,we evaluated the osteogenic effects of h PDLCs with an adenoviral vector encoding humanβ-defensin 3 in an inflammatory microenvironment.Then humanβ-defensin 3 gene-modified rat periodontal ligament cells were transplanted into rats with experimental periodontitis to observe their effects on periodontal bone repair.We found that the humanβ-defensin 3 gene-modified h PDLCs presented with high levels of osteogenesis-related gene expression and calcium deposition.Furthermore,the p38 MAPK pathway was activated in this process.In vivo,humanβ-defensin 3 gene-transfected rat PDLCs promoted bone repair in SD rats with periodontitis,and the p38 mitogen-activated protein kinase(MAPK)pathway might also have been involved.These findings demonstrate that humanβ-defensin 3 accelerates osteogenesis and that humanβ-defensin 3 gene modification may offer a potential approach to promote bone repair in patients with periodontitis.

Sustained delivery of vascular endothelial growth factor mediated by bioactive methacrylic anhydride hydrogel accelerates peripheral nerve regeneration after crush injury

Neurotrophic factors,currently administered orally or by intravenous drip or intramuscular injection,are the main method for the treatment of peripheral nerve crush injury.However,the low effective drug concentration arriving at the injury site results in unsatisfactory outcomes.Therefore,there is an urgent need for a treatment method that can increase the effective drug concentration in the injured area.In this study,we first fabricated a gelatin modified by methacrylic anhydride hydrogel and loaded it with vascular endothelial growth factor that allowed the controlled release of the neurotrophic factor.This modified gelatin exhibited good physical and chemical properties,biocompatibility and supported the adhesion and proliferation of RSC96 cells and human umbilical vein endothelial cells.When injected into the epineurium of crushed nerves,the composite hydrogel in the rat sciatic nerve crush injury model promoted nerve regeneration,functional recovery and vascularization.The results showed that the modified gelatin gave sustained delivery of vascular endothelial growth factors and accelerated the repair of crushed peripheral nerves.