Glycoprotein non-metastatic melanoma protein B (GPNMB), a biomarker of Parkinson's disease (PD), remains at trace levels in clinical samples, which poses a major obstacle for early diagnosis. Herein, a fiber-optic surface plasmon resonance (FO-SPR) sensor based on MOF@Pt-AuPt plasmonic scaffolds is developed for ultrasensitive detection of GPNMB. In-situ growth of Pt nanoparticles on two-dimensional porphyrinic Cu-TCPP MOF forms a Schottky barrier that narrows the optical band gap from 2.53 eV to 2.38 eV, promoting photogenerated charge separation. Subsequent electrostatic assembly of Au@Pt core-shell satellites generates dense localized surface plasmon resonance "hot spots" that strongly couple with the surface plasmon wave on the Au film, and finite-difference time-domain simulations confirm that the interfacial electric field intensity increases from 3.29 to 11.5 (arbitrary units, a.u.). The sensor exhibits excellent refractive index sensing performance, achieving a sensitivity of 3768.64 nm/RIU, which represents a 131.99% improvement over conventional Au-coated optical fibers. For GPNMB detection, the sensor achieves a limit of detection as low as 0.06 pg/mL, more than sevenfold enhancement. Moreover, the sensor is capable of unambiguously discriminating PD patients from healthy controls while exhibiting superior hydrophilicity, antifouling capability, and selectivity. This method provides a practical and versatile detection strategy for the early diagnosis of Parkinson's disease.

PARthanatos is a form of programmed cell death increasingly implicated in neurodegenerative diseases and ischemic stroke. Although classical PARP-1 inhibitors can interrupt this pathway, their prolonged use carries a risk of genomic instability. Screening an NMPA-approved compound library identified compound Bifendate (DDB), a clinical used anti-hepatitis drug, as a effective PARthanatos suppressor. In HeLa and SH-SY5Y cells exposed to the PARthanatos inducer MNNG, DDB increased cell viability by approximately 30 % and 70 %, respectively. This inhibitory effect was not attributable to altered protein levels of PARP-1, AIF, or MIF, but to the blockade of AIF translocation from mitochondria to the cytoplasm and nucleus. Mechanistic analyses revealed that DDB treatment can significantly activate MEK, ERK and then phosphorylate Bad. This activation helps to maintain the mitochondrial membrane potential and permeability, and prevent the release of AIF, and thereby blocks the PARthanatos cascade downstream of PARP-1 activation. Unlike PARP-1 inhibitors, DDB does not interfere with PARP-1 enzymatic activity. Instead, DDB exerts its effects by modulating ERK signaling and enhancing ERK activation. Collectively, these findings provide novel insights into the development of neuroprotective drugs that inhibit PARthanatos without compromising PARP-1 function, highlighting DDB as a promising therapeutic candidate for neurological disorders.

Ischemic stroke currently lacks evidence-based neuroprotective agents, primarily due to the challenge of timely intervention, which often occurs after the onset of irreversible neuronal damage. To address this, this study investigates the PARthanatos pathway, a form of regulated cell death triggered by DNA damage. Utilizing MNNG-induced cellular PARthanatos models, we screened a library of 2,939 traditional Chinese medicine monomers and identified Bruceine E, a natural product derived from Brucea javanica (bitterwood), as a potent inhibitor of PARthanatos at nanomolar concentrations, acting via the inhibition of PARP-1 overactivation. Bruceine E effectively prevents the accumulation of PAR-modified proteins, mitigates mitochondrial membrane potential collapse, and inhibits AIF nuclear translocation. Mechanistically, molecular docking, molecular dynamics simulations, surface plasmon resonance (SPR), and thermal stability assays demonstrate that Bruceine E interacts with the NAD + catalytic pocket of PARP-1 through six hydrogen bonds, exhibiting fast-binding and slow-dissociation kinetics. Furthermore, PARP1 overexpression rescue experiments confirmed that PARP1 overexpression markedly reversed the neuroprotective effect of Bruceine E, indicating that its pharmacological action is specifically dependent on PARP1 regulation. In a permanent distal middle cerebral artery occlusion (pdMCAO) model of C57BL/6 mice, a single intraperitoneal injection of 10 mg/kg Bruceine E administered 4.5 h after occlusion reduced infarct volume by approximately 80.2%, histological evidence confirmed that a single intraperitoneal administration of BE provided effective neuroprotection against ischemic brain injury.

Dihydroxyphenylalanine decarboxylase (DDC) is essential for the synthesis of neurotransmitters and serves as an important biomarker reflecting the progression of Parkinson's disease (PD). Early detection and diagnosis of PD are significantly enhanced by the sensitive and accurate quantification of DDC in serum. This study presents a simple and sensitive electrochemical sandwich sensor for DDC detection in serum, leveraging the unique binding capabilities of lipopolysaccharide binding protein (LBP) and the enhanced electrochemical performance of polyethylenimine (PEI)-stabilized MXene. LBP possessing novel structural domains complementary to DDC was utilized to specifically capture the target analyte. The synergistic combination of these materials enabled the construction of an electrochemical sandwich sensor with highly sensitive and specific for DDC detection. The sensor demonstrated a broad linear detection range from 1 ng/mL to 200 ng/mL, calculating a limit of detection (LOD) as low as 88.96 pg/mL. In addition, the binding affinity of LBP and DDC was verified by molecular docking and surface plasmon resonance (SPR), providing a solid foundation for the subsequent clinical diagnosis of Parkinson's disease. This high-sensitivity detection strategy effectively distinguished between PD patients and healthy individuals, demonstrating its potential as a powerful tool for early detection and clinical analysis of neurological diseases assessment.

Glycoprotein non-metastatic melanoma protein B (GPNMB) is a candidate biomarker of Parkinson’s disease (PD). The development of a novel method for highly sensitive and specific detection of GPNMB is essential for PD’s early and accurate diagnosis. An electrochemical biosensor was developed utilizing silver nanoparticle-loaded mesoporous silica (Ag@MSNs) with 4-mercaptophenylboronic acid (4-MPBA) modification for the sensitive and selective detection of GPNMB. MSNs were utilized as an excellent loading substrate for silver nanoparticles. 4-MPBA at the surface of Ag@MSNs can bind GPNMB specifically through the boronic acid group and multi-glycosylation sites of GPNMB. The finally prepared Ag@MSNs-MPBA exhibited a core–shell structure, which served as a highly efficient electrochemical signal amplifier and selective binding reporter. The enhanced electrochemical signal sensitivity was attributed to the output of the electrochemical signal of Ag through preventing the agglomeration of AgNPs and efficient capture of GPNMB by Ag@MSNs-MPBA. The analysis results demonstrated that the developed electrochemical sensor successfully detected GPNMB with an LOD as low as 0.9011 ng/mL (S/N = 3) in the detection range 1 to 100 ng/mL. The low-cost Ag@MSNs-MPBA replaces the conventional secondary antibody and can integrate signal labels and signal amplification. This work highlights the significant improvement in electrochemical sensing and detection performance achieved by utilizing Ag@MSNs-MPBA, providing a straightforward and general approach for low-cost and sensitive detection of glycoproteins through electrochemical measurement.

Melittin (MEL) is the main bioactive component of bee venom and has been reported to have various pharmacological effects. This study investigates the protective effect of MEL on MPP+-injured HT22 cells and the possible mechanisms involved. We treated the cells with 4 mM MPP+ for 24 h to induce a cellular injury model. HT22 cells were pretreated with 0.1 mu M MEL for 6 h and then exposed to mM MPP+ for 24 h. We measured cell viability, the expression of Bax, the indicators and protein levels associated with apoptosis and parthanatos, and the co-localisation of MEL and mitochondria, and mitochondrial function-related indices such as the mitochondrial membrane potential (MMP) and mito-SOX. We show that PAR protein expression was significantly increased in the MPP+-treated cell model and that the parthanatos inhibitor DPQ significantly reduced MPP+-induced cell death, suggesting that MPP+can cause PARP1-dependent cell death. MEL significantly inhibited cell death, increased cell viability as well as NAD+ and ATP levels, increased the expression of Bcl-2 and suppressed the activation of Bax, cleaved-caspase3, and cleaved-PARP1. Moreover, MEL was found to be localised on the mitochondria of HT22 cells and to improve mitochondrial functions including increased MMP and decreased mitochondrial reactive oxygen species. We speculate that MEL may protect neurons against MPP+-induced HT22 cell injury by inhibiting Bax activation, suppressing changes in mitochondrial permeability, and improving mitochondrial function, thereby preventing cell parthanatos and apoptosis.

IntroductionCerebral ischemic stroke (CIS) is caused by the interruption of cerebral blood circulation due to thrombosis or embolism and is the second-leading cause of mortality worldwide. The neuronal death and motor dysfunction resulting from CIS are primarily attributed to the induction of PARthanatos in neurons at the site of ischemia. Blocking parthanatos is a promising treatment for CIS.MethodsThe effect of medroxyprogesterone treatment on PARthanatos in vitro was examined by CCK8 assay and flow cytometry and the target protein of medroxyprogesterone was then identified by a series of assays, including western blotting, immunofluorescence, cell thermal shift assay and molecular docking. Subsequently, the efficacy of medroxyprogesterone in the treatment of ischemic stroke was evaluated by FJC staining.ResultsIn our study, medroxyprogesterone was able to block the occurrence of PARthanatos in Hela cells induced by MNNG. PARP-1 activity did not change after medroxyprogesterone treatment but prevented MNNG-induced apoptosis inducing factor (AIF) release from mitochondria by improving the stability of phosphorylated extracellular signal-regulated kinase (ERK). In vivo, medroxyprogesterone significantly reduces neuronal death in mouse models of CIS by inhibiting PARthanatos.ConclusionOur findings indicate that medroxyprogesterone effectively inhibits PARthanatos not by affecting the activity of PARP-1, but by directly binding to ERK and stabilizes the active phosphorylated ERK, thereby inhibiting AIF translocation. Furthermore, medroxyprogesterone shows potential as a neuroprotective agent for patients with CIS, potentially enhancing post-stroke recovery and reducing societal burdens.

Cmyc , a proto-oncogene, is expressed at extremely low levels in mature neurons and is traditionally thought to have no function in these cells. However, recent studies suggest that Cmyc may play a crucial role in maintaining the health and function of mature dopaminergic neurons. This study assessed the role of Cmyc in dopaminergic neurons and its significance in Parkinson’s disease. We used a conditional knockout approach to specifically delete Cmyc in substantia nigra dopaminergic neurons of adult mice. Our findings showed that Cmyc deletion led to progressive neuron loss, Parkinson’s disease-like symptoms, downregulation of Klotho, and upregulation of senescence-associated inflammatory factors, along with enhanced oxidative stress and nitrated alpha-synuclein accumulation, ultimately causing neuronal death. In vitro experiments confirmed increased senescence in C-MYC knockout cells, which was partially reversible by KLOTHO overexpression. We conclude that low-level Cmyc expression is essential for maintaining the health of mature dopaminergic neurons and preventing neurodegeneration, and suggest the c-Myc/Klotho axis as a potential therapeutic target for age-related neurodegenerative diseases, including Parkinson’s disease. Our study introduces a novel mouse model for Parkinson’s disease that replicates a condition associated with normal aging, offering a valuable tool for future research into disease mechanisms and therapeutic strategies.

Background Because adverse reactions or drug resistance are often found after current chemotherapies for metastatic colorectal cancer (mCRC), new treatments are still in demand. Shenqi Sanjie Granules (SSG), an antitumor compound preparation of traditional Chinese medicine, has been recognized for its ability in clinical practice of oncotherapy. Nevertheless, the precise effects of SSG in colorectal cancer (CRC) and underlying mechanisms through which SSG inhibits CRC remain uncertain. The current study aimed to evaluate the anti-CRC activity of the Chinese herbal compound preparation SSG and investigate the underlying mechanisms of action. Materials and Methods Initially, nine distinct cancer cell lines, including five CRC cell lines, one breast cancer cell line, two lung adenocarcinoma cell lines and one cervical cancer cell line, were used to evaluate the antitumor activity of SSG, and the mouse CRC cell line CT26 were used for further research. In vitro experiments utilizing diverse assays were conducted to assess the inhibitory effects of the SSG on CT26. Furthermore, subcutaneous syngeneic mouse model and AOM (azoxymethane) / DSS (dextran sodium sulfate) induced in-situ colitis-related mouse CRC model were used to evaluate the antitumor potential and biotoxicity of SSG in vivo. To elucidate the underlying molecular mechanisms, transcriptome sequencing and network pharmacology analysis were performed. Meanwhile, verification is carried out with quantitative real-time PCR (qRT-PCR) and flow cytometry (FCM) analysis. Results Our in vitro inhibition study showed that SSG could effectively inhibit CRC cell line CT26 growth and metastasis, and induce cell death. Neither of apoptosis inhibitor, necroptosis inhibitor, ferroptosis inhibitor, but the combination of the three diminished SSG-induced cell death, suggesting that multiple cell death pathways were involved. Both the syngeneic CRC model and the in-situ CRC model indicated SSG inhibited CRC in vivo with few toxic side effects. Further mechanistic study suggested SSG treatment activated the ferroptosis pathway, particularly mediated by Hmox1, which was upregulated scores of times. Network pharmacology analysis indicated that the active ingredients of SSG, including Quercetin, Luteolin and Kaempferol were potential components directly upregulated Hmox1 expression. Conclusions Collectively, our findings indicate that the administration of SSG has the potential to inhibit CRC both in vitro and in vivo. The mechanism by which this compound preparation exerts its action is, at least partly, the induction of ferroptosis through upregulating Hmox-1 by its three active ingredients Quercetin, Luteolin and Kaempferol.

To combat the SARS-CoV-2 pandemic, innovative prevention strategies are needed, including reducing ACE2 expression on respiratory cells. This study screened approved drugs in China for their ability to downregulate ACE2. Daphnetin (DAP) was found to significantly reduce ACE2 mRNA and protein levels in PC9 cells. DAP exerts its inhibitory effects on ACE2 expression by targeting HIF-1α and JAK2, thereby impeding the transcription of the ACE2 gene. The SARS-CoV-2 pseudovirus infection assay confirmed that DAP-treated PC9 cells exhibited decreased susceptibility to viral infection. At therapeutic doses, DAP effectively lowers ACE2 expression in the respiratory systems of mice and humans. This suggests that DAP, already approved for other conditions, could be a new preventive measure against SARS-CoV-2, offering a cost-effective and accessible way to reduce SARS-CoV-2 spread.

This study investigated the feasibility of polypyrimidine tract binding protein 1 knockout mouse spinal cord astrocytes in the treatment of spinal cord injury. We cultured primary astrocytes from the spinal cord of newborn mice, then integrated sh polypyrimidine tract binding protein 1 into the genome by lentiviral transduction and verified their knockout efficiency. Immunofluorescence staining and transcriptome sequencing were performed to test whether the cells differentiated into neurons. The results showed that polypyrimidine tract binding protein 1 knockout could transform spinal cord astrocytes into functional neurons in one step. After whole transcriptome sequencing analysis, gene ontology annotation and Kyoto encyclopedia of genes and genomes pathway enriched differentially expressed genes revealed that mitogen-activated protein kinase, erythroblastic oncogene B and Ras-associated protein-1 signaling pathways may be involved in this transdifferentiating process. Predictive analysis of microRNA target genes revealed microRNAs that may be involved in neural differentiation, as well as long noncoding RNA and circular RNA that indirectly regulate transdifferentiating through these microRNAs. Analysis of the circular RNA-microRNA-messenger RNA network revealed three circular RNA that may play a key role in transdifferentiating, as well as possible regulatory mechanisms in this process. We further constructed a mouse spinal cord injury model and found that 9 w after surgery, the basso mouse scale score of the right hind limb of the sh polypyrimidine tract binding protein 1 lentivirus injection group was significantly higher than that of the scrambled lentivirus injection group. Our results suggest that polypyrimidine tract binding protein 1 knockdown of reactively proliferating spinal cord astrocytes in spinal cord injury promotes neuron generation.

The oncogenic role of Ladinin-1 (LAD1), an anchoring filament protein, is largely unknown. In this study, we conducted a series of studies on the oncogenic role of LAD1 in lung adenocarcinoma (LUAD). Firstly, we analyzed the aberrant expression of LAD1 in LUAD and its correlation with patient survival, tumor immune infiltration, and the activation of cancer signaling pathways. Furthermore, the relationship between LAD1 expression and K-Ras and EGF signaling activation, tumor cell proliferation, migration, and colony formation was studied by gene knockout/knockout methods. We found that LAD1 was frequently overexpressed in LUAD, and high LAD1 expression predicts a poor prognosis. LAD1 exhibits promoter hypomethylation in LUAD, which may contribute to its mRNA upregulation. Single-sample gene set enrichment analysis (ssGSEA) showed that acquired immunity was negatively correlated with LAD1 expression, which was verified by the downregulated GO terms of "Immunoglobulin receptor binding" and "Immunoglobulin complex circulating" in the LAD1 high-expression group through Gene Set Variation Analysis (GSVA). Notably, the Ras-dependent signature was the most activated signaling in the LAD1 high-expression group, and the phosphorylation of downstream effectors, such as ERK and c-jun, was strongly inhibited by LAD1 deficiency. Moreover, we demonstrated that LAD1 depletion significantly inhibited the proliferation, migration, and cell-cycle progression of LUAD cells and promoted sensitivity to Gefitinib, K-Ras inhibitor, and paclitaxel treatments. We also confirmed that LAD1 deficiency remarkably retarded tumor growth in the xenograft model. Conclusively, LAD1 is a critical prognostic biomarker for LUAD and has potential as an intervention target.

Brain natriuretic peptide (BNP) has been a disease marker in the diagnosis of heart failure. In this study, gold nanoparticles modified with Hemin (H-AuNPs) as nanozymes were used to oxidize ABST and MB to amplified colorimetric and electrochemical redox signals respectively. BNP was combined with H-AuNPs (BNP-H-AuNPs) through electrostatic adsorption to construct competitive nanozyme probes. Target BNP in the sample compete with BNP-H-AuNPs to bind the antibody-modified magnetic nanoparticles (AntiBNP-MNPs). Due to the excellent catalytic performance of the nanozyme, BNP can be observed well by colorimetric and electrochemical assays. Electrochemical method ensured more accurate detection of BNP with a wide detection range (1-200 pg/mL) and a low detection of limit (0.03 pg/mL). Meanwhile, the results of the experiment can be easily observed with the naked eye by simple colorimetric method with a range from 5 ng/mL to 25 ng/mL and a limit of detection down to 80.3 pg/mL. Thus, based on the important role of H-AuNPs, this assay has exhibited potential value of detection the other small proteins through this competitive nanozyme method.

Nitrated α-syn (nitro-α-syn) is a biomarker for Parkinson's desease (PD), and its sensitive detection in serum is of great importance for early PD diagnosis. Silver-coated copper MOF (Cu-MOF@Ag) with outstanding oxidase activity and electrochemical response property was designed and synthesized. Cu-MOF@Ag exhibited excellent oxidase activity with a low Km value (0.568 mM), avoiding the addition of strong oxidant to catalyze chromogenic substrate, which enhanced the colorimetric stability. Silver nanoparticles as an electrochemical signal reporter can be easily decorated on the surface of Cu-MOF with bifunctional groups (-SH and -NH2) material, which can increase the electrochemical signal output. The α-syn antibody modified Cu-MOF@Ag and nitro-α-syn modified magnetic nanoparticle were used as immunoprobes to specifically capture nitro-α-syn. A dual-modal immunosensor was fabricated for the simple and reliable detection of nitro-α-syn based on Cu-MOF@Ag. Combing colorimetric and electrochemical detection, nitro-α-syn can be determined quantitatively within a wide linear range (10–350 ng/mL) with low detection limit (0.5 ng/mL). The ability of the sensor with magnetic separation and dual signal analysis allowed to successfully detect nitro-α-syn and distinguish PD patients from healthy people (P < 0.005). Thanks to its excellent selectivity, stability, and the precision of 2.69

Tex264 is an endoplasmic reticulum (ER) membrane protein that was recently demonstrated to act as an ER-phagy receptor under starvation conditions to mediate endoplasmic reticulum autophagy. However, how Tex264 functions in the central nervous system (CNS) and tumors is unclear. Here, we identified 89 proteins from the rat brain that may specifically interact with Tex264 and confirmed the interaction between sorting nexin 27 (SNX27) and Tex264 by coimmunoprecipitation and immunofluorescence. Our results indicated that Tex264 may promote recycling of membrane proteins from endosomes to the cell plasma membrane by recruiting SNX27 retromer vesicles. siRNA-mediated knockdown of TEX264 in HeLa cells did not affect cell proliferation but did significantly inhibit cell migration through a mechanism that may involve a reduction in SNX27-mediated Itgα5 receptor membrane recycling. Results of this study helped identify potential binding Tex264 partners and provide insights into Tex264 functions in the CNS and in tumors.

Abstract Introduction The role of extra-hypothalamic thyrotropin-releasing hormone (TRH) has been investigated by pharmacological studies using TRH or its analogues and found to produce a wide array of effects in the central nervous system. Methods Immunofluorescence, In situ labeling of DNA (TUNEL), in situ hybridization chain reaction and quantitative real-time polymerase chain reaction were used in this study. Results We found that the granular cells of the dentate gyrus expressed transiently a significant amount of TRH-like immunoreactivity and TRH mRNA during the 6–24 h period following global cerebral ischemia/reperfusion injury. TUNEL showed that apoptosis of neurons in the CA1 region occurred from 48 h and almost disappeared at 7 days. TRH administration 30 min before or 24 h after the injury could partially inhibit neuronal loss, and improve the survival of neurons in the CA1 region. Conclusion These data suggest that endogenous TRH expressed transiently in the dentate gyrus of the hippocampus may play an important role in the survival of neurons during the early stage of ischemia/reperfusion injury and that delayed application of TRH still produced neuroprotection. This delayed application of TRH has a promising therapeutic significance for clinical situations.

Alzheimer's disease (AD) is one of the most prevalent neurodegenerative disease. Plasma total phosphorylated tau (P-tau) with multi-phosphorylation sites is a potentially efficient biomarker for the diagnosis and prognosis of AD. Single phosphorylation site recognition methods lead to the false positive detection of P-tau and decrease the accuracy of AD analysis. In this work, a simple and precise electrochemical immunoassay was developed for the specific detection of P-tau. Calixarene derivative ProLinker was utilized to orient Anti-tau onto the surface of gold electrode. ProLinker provide abundant binding sites for immobilized antibodies via host-guest recognitions and ensure Anti-tau activity through non-covalent interactions. Flower-shaped TiO2 (F-TiO2) forming coordination bonds with the phosphate group of phosphorylated protein were utilized as signal amplification tags to construct a simple sandwich assay for multiple phosphorylated sites sensing of P-tau. The combination of F-TiO2 and Anti-tau ensured the bispecific and precise recognition of P-tau with a wide detection range (1-200 ng/mL) and a low detection of limit (1.774 pg/mL). With large specific surface area, high electron hinder ability, and great stability, F-TiO2 as an amplification molecule can improve the performance of the immunosensor and produce sensitive signal changes according to the concentration of P-tau in the clinical samples. Hence, this P-tau immunoassay paves the way for the development of efficient detection tools for clinical diagnosis and monitoring.

How to use bioinformatics methods to quickly and accurately locate the effective targets of traditional Chinese medicine monomer (TCM) is still an urgent problem needing to be solved. Here, we used high-throughput sequencing to identify the genes that were up-regulated after cells were treated with TCM monomers and used bioinformatics methods to analyze which transcription factors activated these genes. Then, the binding proteins of these transcription factors were analyzed and cross-analyzed with the docking proteins predicted by small molecule reverse docking software to quickly and accurately determine the monomer’s targets. Followeding this method, we predicted that the TCM monomer Daphnoretin (DT) directly binds to JAK2 with a binding energy of −5.43 kcal/mol, and activates the JAK2/STAT3 signaling transduction pathway. Subsequent Western blotting and in vitro binding and kinase experiments further validated our bioinformatics predictions. Our method provides a new approach for quickly and accurately locating the effective targets of TCM monomers, and we also have discovered for the first time that TCM monomer DT is an agonist of JAK2.

PER1 is a core component of the internal time-keeping system. In the suprachiasmatic nucleus, it serves as the primary circadian pacemaker in mammalian brains. PER1 functions with other clock components to generate a feedback loop involving the transcriptional repression of gene expression to produce a circadian rhythm with an approximately 24-hour cycle. Post-transcriptional modifications (PTMs) are a basic regulatory mechanism that both perpetuate self-sustained oscillations and interpret metabolic input into circadian physiology by affecting factors such as protein stability, interactions, localization, and activity. Here we examined whether the serine/threonine protein kinase WNK3, which is expressed in a circadian rhythm, can interact and colocalize with PER1 in the SCN. In rats, WNK3 knockdown in the SCN is associated with altered sleep patterns. Moreover, WNK3 can phosphorylate PER1 to promote its degradation and is associated with circadian oscillations when PER1 is expressed in vitro.