Pathogens across kingdoms converge on hijacking plant immune hubs, but why certain host proteins emerge as evolutionary "hotspots" remains elusive. Here, we reveal that the transcription factor TCP14, a prime effector target, serves as a circadian-immune integrator. The clock components CCA1 and ZTL orchestrate TCP14 levels through transcriptional activation and proteasomal degradation, respectively, generating diurnal oscillations in TCP14 activity. Reciprocally, TCP14 promotes the transcription of several clock genes by directly binding to their promoters. Time-course RNA-seq and genetic analyses demonstrated that TCP14 plays a broad role in regulating rhythmic gene expression, hypocotyl growth, and time-of-day-specific defense responses. Moreover, a pathogen effector, HaRxL21, directly interacts with TCP14 and transcriptional repressors TPL/TPR1 to suppress defense genes, perturbing time-gated defense. Our work establishes TCP14 as a chrono-immune nexus whose vulnerability arises from its role in coupling circadian timing to defense prioritization, offering a paradigm for how pathogens disrupt host rhythms through hub sabotage.

We previously reported that integrin alpha 7 (ITGA7) was downregulated in colorectal cancer (CRC) tissues and CRC cell lines and that the lower expression of ITGA7 in CRC tissues was correlated with distant metastasis, suggesting that ITGA7 may function as a suppressor in CRC. The present research was conducted to further investigate the role and mechanisms of ITGA7 in CRC progression. First, bisulfite modification and genomic sequencing (BSP) results showed that the methylation rate of ITGA7 promoter was higher in 10 CRC tissues than in the matched normal tissues. Additionally, 5-Aza-CdR treatment increased ITGA7 expression in CRC cells. Gain-of-function assays revealed the inhibitory role of ITGA7 in CRC cell proliferation and migration. Mechanistically, RNA sequencing, RT-qPCR, and cytoplasm and nuclear separation and rescue assays indicated that knockdown of ITGA7 activated the transcription of MMP9, SETD7, and ADAM15 by enhancing the nuclear translocation of NF-κB. Moreover, CoIP and Western blot suggested a mechanistic model in which ITGA7 binds to CKAP4 to block the interaction of CKAP4 and PI3K p85α and thereby suppress the PI3K/AKT/NF-κB pathway. Accordingly, the current study suggests that ITGA7 functions as a suppressor in CRC progression and that its expression is controlled by promoter methylation.

CXXC5, a member of the CXXC family of zinc-finger proteins, is associated with numerous pathological processes. However, the pathophysiological function of CXXC5 has not been clearly established. Herein, we found that CXXC5 interacts with the CRL4B and NuRD complexes. Screening of transcriptional targets downstream of the CXXC5-CRL4B- NuRD complex by next-generation sequencing (chromatin immunoprecipitation sequencing) revealed that the complex regulates the transcriptional repression process of a cohort of genes, including TSC1 (tuberous sclerosis complex subunit 1), which play important roles in cell growth and mammalian target of rapamycin signaling pathway regulation, and whose abnormal regulation results in the activation of programmed expression increased after stimulation with vitamin B2 but decreased after vitamin D treatment. We also found that the CXXC5-CRL4B-NuRD complex promotes the proliferation of tumor cells in vitro and accelerates the growth of breast cancer in vivo. The expression of CXXC5, CUL4B, and MTA1 increased during the occurrence and development of breast cancer, and correspondingly, TSC1 expression decreased. Meanwhile, a high expression of CXXC5 was positively correlated with the histological grade of high malignancy and poor survival of patients. In conclusion, our study revealed that CXXC5-mediated TSC1 suppression activates the mammalian target of rapamycin pathway, reduces autophagic cell death, induces PD-L1-mediated immune suppression, and results in tumor development, shedding light on the mechanism of the pathophysiological function of CXXC5.

In recent years, the potassium voltage-gated channel subfamily D (KCND) channels, particularly KCND2 (also known as Kv4.2), have been suggested to play a role in a variety of cancers, but their role in breast cancer has not yet been revealed. We analyzed RNA sequencing data from The Cancer Genome Atlas database and the Genotype-Tissue Expression database to investigate the differential expression of KCND2 in breast cancer and normal breast tissue. In addition, we leveraged GO and KEGG analysis techniques to gain a better understanding of the potential functional enrichment of 500 genes related to KCND2. Our findings were validated using collected tissue samples and clinical data from hospitals showed that KCND2 is a crucial independent factor in the prognosis of breast cancer patients. The higher the expression of KCND2, the shorter the survival time of breast cancer patients. Colony formation assay confirmed that KCND2 promotes the proliferation of breast cancer cells, whereas transwell assay and wound healing assay verified that KCND2 promoted breast cancer invasion and migration. In addition, 5-Ethynyl-2′-deoxyuridine (EdU) and flow cytometry revealed that KCND2 affected the cycle changes of breast cancer cells and contributed to the G1/S phase transition of breast cancer cells. Overall, our study demonstrates that KCND2 holds a promising potential as a significant target for breast cancer diagnosis and therapy.

Background: Although the management of the lumbar disease is highly dependent on the severity of the patient’s condition, optimal surgical techniques to reduce the risk of adjacent degeneration disease (ADS) remain elusive. Based on in vitro biomechanical tests of the cadaver spine, this study aimed to comparatively analyze the kinematic responses of the spine with dynamic and rigid fixations (i.e., Coflex fixation and posterolateral fusion) after single-or double-level lumbar fusion in daily activities. Methods: Six human lumbar specimens (L1-S1) were selected for this experiment, and the sagittal parameters of each lumbar specimen were measured in the 3D model. The specimens were successively reconstructed into five groups of models: intact model, single-level L4-5 Coflex fixation model, single-level L4-5 Fusion (posterior pedicle screw fixation) model, double-level L4-5 Coflex + L5-S1 Fusion model; and double-level L4-5 Fusion + L5-S1 Fusion model. The pure moment was applied to the specimen model to simulate physiological activities in daily life through a custom-built robot testing device with an optical tracking system. Results: For single-level lumbar fusion, compared to the traditional Fusion fixation, the Coflex dynamic fixation mainly restricted the extension of L4-L5, partially retained the range of motion (ROM) of the L4-L5 segment, and reduced the motion compensation of the upper adjacent segment. For the double-level lumbar fixation, the ROM of adjacent segments in the Coflex + Fusion was significantly decreased compared to the Fusion + Fusion fixation, but there was no significant difference. In addition, PT was the only sagittal parameter of the preoperative lumbar associated with the ROM under extension loading. The Coflex fixation had little effect on the original sagittal alignment of the lumbar spine. Conclusion: The Coflex was an effective lumbar surgical technique with a less altering kinematic motion of the lumbar both at the index segment and adjacent segments. However, when the Coflex was combined with the fusion fixation, this ability to protect adjacent segments remained elusive in slowing the accelerated degradation of adjacent segments.

LINC00472 is reported to play a role in suppressing tumors in cancers such as lung cancer and hepatocellular carcinoma, among others. We made investigations into the effects of LINC00472 in oral squamous cell carcinoma (OSCC) progression to explore the underlying molecular mechanisms. By qRT-PCR, we assessed the LINC00472 expression in OSCC tissues and cells and performed functional analysis to investigate how LINC00472/miR-455-3p/ELF3 impacts OSCC cell proliferation, apoptosis, and cell cycle. The role that LINC00472 plays in OSCC tumor growth was examined by establishing a xenograft model. Down-regulation of LINC00472 occurred in tissues and cells of an OSCC tumor. LINC00472 overexpression caused OSCC cell proliferation to be inhibited, cell apoptosis to be promoted, and cell cycle arrest to be induced. As a competing endogenous RNA (ceRNA), LINC00472 can block miR-455-3p function and further promote ELF3 expression. The overexpression of miR-455-3p or ELF3 knockdown was shown to be capable of reversing the anti-tumor effects of LINC00472 in OSCC. In vivo experiments confirmed the tumor-suppressing role of LINC00472 in the progression of OSCC. In short, we found that the novel LINC00472 inhibits OSCC growth via the miR-455-3p/ELF3 axis. LINC00472 and its targeted miR-455-3p/ELF3 axis may represent valuable targets for treating OSCC.

Abstract Background: The effectiveness of comprehensive geriatric assessment (CGA) has been confirmed to reduce perioperative complications of geriatric patients. But there is no universally accepted standardization of CGA for orthopedic surgery. In this study, a novel CGA strategy was applied to evaluate the conditions of elderly patients undergoing orthopedic surgery from a broad view and to identify potential risk factors for postoperative complications. Methods: A prospective cohort study was conducted from March 2019 to December 2020.The study enrolled patients for elective or confined orthopedic surgery if they were over the age of 75 years old. All patients were enrolled and treated with help from a multidisciplinary team. A structured CGA was conducted to identify high-risk elderly patients and to facilitate coordinated multidisciplinary team care by a geriatric team. The basic patient characteristics and CGA results were collected in addition to postoperative complication and mortality rates. Multivariate logistic regression analysis was used to identify risk factors for postoperative complications. Results: A total of 214 patients with an age of 81.07 ± 4.78 (range, 75–100) years were prospectively enrolled in this study. In total, 66 (30.8%) complications were registered, including one death from myocardial infarction (mortality rate, 0.5%). Poor ADL and IADL were accompanied by frailty, worse American Society of Anesthesiologists (ASA) score, visual analogue scale score, and nutritional status. Poor ADL was also associated with higher risks of falling, polypharmacy, and cardiac and respiration complications. Poor IADL was associated with higher risk of cardiac and respiration complications. Higher stroke risk was accompanied by higher risks of cardiac complications, delirium, and hemorrhage. Worse ASA scale was associated with worse ADL, IADL, and frailty, and higher delirium risk. Multivariate logistic regression analysis showed that spinal fusion (odds ratio [OR], 0.73; 95% confidence interval [CI], 0.65 to 0.83; p = 0.0214), blood loss(OR, 1.68; 95% CI, 1.31 to 2.01; p = 0.0168), ADL (severe dysfunction or worse) (OR, 1.45; 95% CI, 1.16 to 1.81; p = 0.0413), IADL (serious dependence) (OR, 1.08; 95% CI, 1.33 to 1.63; p = 0.0436), renal function (CKD ≥ stage 3a) (OR, 2.01; 95% CI, 1.54 to 2.55; p = 0.0133), and malnutrition(OR, 2.11; 95% CI, 1.74 to 2.56; p = 0.0101) were independent risk factors for postoperative complications. All P <0.05.Conclusion: Spinal fusion, blood loss, ADL (severe dysfunction or worse), IADL (serious dependence), renal function (CKD ≥ stage 3a) and nutrition MNA (malnourished) were independent risk factors of postoperative complications following orthopaedic surgery in elderly patients.

Multiple causes may contribute to osteoporosis, characterized by a loss in bone mass and density as a consequence of the degradation of bone microstructure and a resultant rise in bone fragility. Recently, increasing attention has been paid to the role of necroptosis in the development of osteoporosis. Necroptosis is orchestrated by a set of proteins known as receptor-interacting protein kinase (RIPK)1, RIPK3, and mixed lineage kinase domain-like protein (MLKL). A necrosome is formed by MLKL, RIPK1, RIPK3, and RIPK3-RIPK3. A dissociated MLKL forms pores in the plasma membrane and eventually leads to necroptosis after translocating from the necrosome. In this review, we discuss a detailed understanding of necroptosis and its associated processes, a better understanding of its interactions with osteoclasts, osteoblasts, and osteocytes, and the associations between necroptosis and diabetic osteoporosis, steroid-induced osteoporosis, and postmenopausal osteoporosis. In addition, a variety of experimental medicines capable of modulating crucial necroptosis processes are highlighted. It's important to note that this is the first review paper to consolidate current data on the role of necroptosis in osteoporosis, and it offers fresh hope for the future treatment of this disease.

Receptor-interacting protein kinase 3 (RIP3) is a critical initiator in mediating necroptosis induced by tumor necrosis factor alpha (TNFα) in L929 cells, so knockdown of RIP3 inhibits TNFα-induced L929 cell necroptosis. However, RIP3 knockdown was shown to switch TNFα-induced necroptosis to apoptosis in L929 cells in other studies. Therefore, whether RIP3 knockdown blocks the TNFα-induced death of L929 cells is controversial. In this study, TNFα activated caspase pathway and induced cell death in RIP3 knockdown L929 cells, and the RIP3-independent cell death had been blocked by Z-VAD-FMK (pan-caspase inhibitor) or caspase 8 knockdown, demonstrating that RIP3 knockdown switched TNFα-induced necroptosis to caspase-dependent apoptosis. Although both TNF receptor type 1-associated death domain protein (TRADD) and RIP1 have been reported to mediate TNFα-induced apoptosis, the knockdown of TRADD, but not RIP1, suppressed TNFα-induced activation of the caspase pathway and subsequent apoptosis in RIP3 knockdown L929 cells. In addition, TRADD bound and activated caspase 8 during the RIP3-independent apoptosis process, indicating that TRADD initiates RIP3-independent apoptosis by activating the caspase pathway. Collectively, we identified the target and mechanism underlying RIP3-independent apoptosis and elucidated the coordinated roles of RIP3 and TRADD in mediating the programmed cell death of L929 cells following TNFα stimulation.

Objective To identify the role of phosphatidylinositol-3-kinase(PI3K) in mediating necroptosis induced by tumor necrosis factor alpha (TNFα) and the involved mechanism.Methods Knockdown of p110α,receptor-interacting protein 1(RIP1) or both p110αand RIP1 was mediated by the specific short hairpin RNA (shRNA) lentivirus and verified by RT-PCR or Western blotting .In addition , Western blotting was used to detect phosphorylation of mixed lineage kinase domain-like protein(MLKL) and protein kinase B(AKT) or tetramerization of MLKL.Cell death was measured by micros-copy and flow cytometry.Results AKT phosphorylation and TNFα-induced necroptosis of L929 cells were suppressed by the inhibitors of PI3K or AKT, as well as p110αknockdown.Moreover, RIP1 knockdown did not inhibit L929 cell death induced by TNFαplus Z-VAD, but the RIP1-independent necroptosis was inhibited by p 110αknockdown.In addition, p110αknockdown suppressed MLKL phosphorylation and tetramerization induced by TNFαwith Z-VAD in L929 cells. Conclusion PI3K mediates necroptosis of L929 cells induced by TNFαby activating AKT and MLKL, respectively.

Propofol can be protective against various insults in many systems. However, the exact mechanisms of propofol protection are still unknown. In the present study, we showed that propofol decreased p53 level through upregulating miR-6983-5p. This might be a possible mechanism of propofol promoting cell survival. In addition, propofol prevented neurons against staurosporine, etoposid, glutamate and serum deprivation treatments. Our study provides a possible mechanism that propofol regulating p53 level through miRNA to prevent cell death.

Macrophage cholesterol efflux is important in maintaining cellular lipid homeostasis and preventing the formation of lipid‑laden foam cells. Although radioactive [3H]‑cholesterol is widely used as a tracer in cholesterol efflux assays, the lengthy and labor‑intensive assay procedure, and the radioactivity disposal procedure limit the use of this assay for high‑throughput screening. In the present study, a novel procedure using fluorescent N‑(7‑nitrobenz‑2‑oxa‑1,3‑diazol‑4‑yl)amino)‑23,24‑bisnor‑5‑xholen‑3β‑ol (NBD)‑cholesterol was developed as a substitute for [3H]‑cholesterol for the measurement of cholesterol efflux in THP‑1‑derived macrophages. NBD‑cholesterol uptake and metabolism in the THP‑1 cells were characterized using fluorescent microscopy and spectrophotometry. NBD‑cholesterol distributed rapidly into the cell organelles, with the exception of the nucleus. The uptake of NBD‑cholesterol in the THP‑1 macrophages was concentration‑ and time‑dependent, and reached a plateau following 4 h incubation. The present study subsequently investigated whether NBD‑cholesterol efflux was correlated with [3H]‑cholesterol efflux in THP‑1 derived macrophages and in human peripheral blood mononuclear cells (PBMCs). The results demonstrated that the percentage of efflux of NBD‑cholesterol in the THP‑1 cells was significantly correlated with that of [3H]‑cholesterol, at various concentrations of HDL or apoA‑1 as lipid acceptors (R2=0.876 for HDL; R2=0.837 for apoA‑1; P<0.001). In the PBMCs, NBD‑cholesterol efflux also correlated significantly with [3H]‑cholesterol efflux (R2=0.887 for HDL; R2=0.872 for apoA‑1; P<0.001). Furthermore, NBD‑cholesterol efflux in the THP‑1 cells exhibited a similar trend to that obseved in the PBMCs. In conclusion, the results of the present study suggested that fluorescent NBD‑cholesterol can be used as a sensitive and specific probe in cholesterol efflux assays in THP‑1‑derived macrophages.