Fine mapping and functional validation of the maize nicosulfuron-resistance gene CYP81A9.

Nicosulfuron, a widely utilized herbicide, is detrimental to some maize varieties due to their sensitivity. Developing tolerant varieties with resistance genes is an economical and effective way to alleviate phytotoxicity. In this study, map-based cloning revealed that the maize resistance gene to nicosulfuron is Zm00001eb214410 (CYP81A9), which encodes a cytochrome P450 monooxygenase. qRT- PCR results showed that CYP81A9 expression in the susceptible line JS188 was significantly reduced compared to the resistant line B73 during 0-192 hours following 80 mg/L nicosulfuron spraying. Meanwhile, a CYP81A9 overexpression line exhibited normal growth under a 20-fold nicosulfuron concentration (1600 mg/L), while the transgenic acceptor background material Zong31 did not survive. Correspondingly, silencing CYP81A9 through CRISPR/Cas9 mutagenesis and premature transcription termination mutant EMS4-06e182 resulted in the loss of nicosulfuron resistance in maize. Acetolactate Synthase (ALS), the target enzyme of nicosulfuron, exhibited significantly reduced activity in the roots, stems, and leaves of susceptible maize post-nicosulfuron spraying. The CYP81A9 expression in the susceptible material was positively correlated with ALS activity in vivo. Therefore, this study identified CYP81A9 as the key gene regulating nicosulfuron resistance in maize and discovered three distinct haplotypes of CYP81A9, thereby laying a solid foundation for further exploration of the underlying resistance mechanisms.

Necrotizing pneumonia associated with macrolide-resistant Mycoplasma in a child.

We reported a pediatric case of necrotizing pneumonia due to macrolide-resistant Mycoplasma pneumoniae, an uncommon presentation of a common disease. Acquisition of resistance does not increase virulence, but it leads to more difficult treatment and potential complications. Macrolide-resistant M. pneumoniae requires extended antibiotic therapy with the addition of a second-line agent and an immunomodulator to promote clinical improvement with minimal sequelae.

Wine- and stir-frying processing of Cuscutae Semen enhance its ability to alleviate oxidative stress and apoptosis via the Keap 1-Nrf2/HO-1 and PI3K/AKT pathways in H2O2-challenged KGN human granulosa cell line.

BACKGROUND: Cuscutae Semen (CS) has been prescribed in traditional Chinese medicine (TCM) for millennia as an aging inhibitor, an anti-inflammatory agent, a pain reliever, and an aphrodisiac. Its three main forms include crude Cuscutae Semen (CCS), wine-processed CS (WCS), and stir-frying-processed CS (SFCS). Premature ovarian insufficiency (POI) is a globally occurring medical condition. The present work sought a highly efficacious multi-target therapeutic approach against POI with minimal side effects. Finally, it analyzed the relative differences among CCS, WCS and SFCS in terms of their therapeutic efficacy and modes of action against H2O2-challenged KGN human granulosa cell line. METHODS: In this study, ultrahigh-performance liquid chromatography (UPLC)-Q-ExactiveTM Orbitrap-mass spectrometry (MS), oxidative stress indices, reactive oxygen species (ROS), Mitochondrial membrane potential (MMP), real-time PCR, Western blotting, and molecular docking were used to investigate the protective effect of CCS, WCS and SFCS on KGN cells oxidative stress and apoptosis mechanisms. RESULTS: The results confirmed that pretreatment with CCS, WCS and SFCS reduced H2O2-induced oxidative damage, accompanied by declining ROS levels and malondialdehyde (MDA) accumulation in the KGN cells. CCS, WCS and SFCS upregulated the expression of antioxidative levels (GSH, GSH/GSSG ratio, SOD, T-AOC),mitochondrial membrane potential (MMP) and the relative mRNA(Nrf2, Keap1, NQO-1, HO-1, SOD-1, CAT). They inhibited apoptosis by upregulating Bcl-2, downregulating Bax, cleaved caspase-9, and cleaved caspase-3, and lowering the Bax/Bcl-2 ratio. They also exerted antioxidant efficacy by partially activating the PI3K/Akt and Keap1-Nrf2/HO-1 signaling pathways. CONCLUSIONS: The results of the present work demonstrated the inhibitory efficacy of CCS, WCS and SFCS against H2O2-induced oxidative stress and apoptosis in KGN cells and showed that the associated mechanisms included Keap1-Nrf2/HO-1 activation, P-PI3K upregulation, and P-Akt-mediated PI3K-Akt pathway induction.

cGAS Deficiency Regulates the Phenotypic Polarization and Glycolysis of Microglia Through Lactylation in Hypoxic-Ischemic Encephalopathy Cell Model.

Promoting the M2 phenotype polarization of microglia is of great significance in alleviating hypoxic-ischemic encephalopathy (HIE). The umbilical artery blood sample was collected to evaluate the expression of cGAS, and the aberrant expressed cGAS was verified in the oxygen glucose deprivation (OGD) microglia which was established to mimic HIE in vitro. Then the regulating role of cGAS on the transformation of microglia M2 phenotype polarization and glycolysis was investigated. Moreover, the lactylation of cGAS in OGD treated microglia was evaluated by western blot. cGAS was found to be highly expressed in umbilical artery blood of HIE group, and OGD treated microglia. OGD interference activated microglia into M1 phenotype by enhancing CD86 and suppressing CD206 levels; meanwhile, the microglia in OGD group highly expressed IL-1ß, iNOS and TNF-α, and lowly expressed IL-4, IL-10, and Arg-1. Inhibition of cGAS promotes the transformation of microglia from M1 to M2 phenotype. Meanwhile, OGD increased ECAR and decreased OCR to regulate glycolysis, cGAS deficiency inhibits glycolysis in OGD treated microglia. Moreover, the pan lysine lactylation (Pan-Kla) levels and lactated cGAS levels in microglia were upregulated in the OGD group. Lactate reversed the effects of cGAS knockdown on microglia polarization and glycolysis. The present study reveals that the cGAS-mediated neuron injury is associated with high level of cGAS lactylation. Inhibition of cGAS promotes the M2 phenotype polarization of microglia and suppress glycolysis. Thereby, targeting cGAS provides a new strategy for the development of therapeutic agents against HIE.

Comparison of proprioception recovery following anterior cruciate ligament reconstruction using an artificial graft versus an autograft.

BACKGROUND: To compare proprioception recovery after anterior cruciate ligament reconstruction (ACLR) with a hamstring tendon autograft versus the artificial Ligament Advanced Reinforcement System (LARS). MATERIAL AND METHODS: Forty patients (9 females, 31 males) with anterior cruciate ligament (ACL) rupture were enrolled in this prospective study. Patients were randomized to two groups, 1) ACLR using a hamstring tendon autograft (n = 20) or 2) ACLR using artificial LARS (n = 20). Proprioception was assessed with knee joint position sense (JPS) passive-passive test at 45° and 75° flexions, with the contralateral healthy knee as a control baseline to calculate the JPS error. Knee JPS absolute error was used as the main outcome variable and defined as the absolute difference between the reproduction and target angles. RESULTS: JPS error in both groups at 3 months after ACLR was significantly higher than that at 12 months. However, no significant difference in JPS error was detected between the LARS and autograft groups at either 3 or 12 months after ACLR. Analyzing JPS data by grouping patients according to whether ACLR was performed more or less than 1 year following injury regardless of graft type showed a statistically significant difference between the groups at 3 months, but not at 12 months, after ACLR. Patients receiving the graft within 1 year of injury had a lower JPS error than those receiving the graft more than 1 year after injury at 3 months. No complications were associated with either ACLR method. CONCLUSION: ACLR with a hamstring tendon autograft or LARS artificial graft is similarly safe and effective for recovering knee proprioception.

3D-printed hemipelvic prosthesis combined with a dual mobility bearing in patients with primary malignant neoplasm involving the acetabulum: clinical outcomes and finite element analysis.

BACKGROUND: Limb salvage reconstruction for pelvic tumors, especially periacetabular tumors, is challenging. We combined the use of dual mobility bearing and 3D-printed hemipelvic prosthesis to improve function and reduce the probability of complications after hemi-pelvic resection in patients with primary acetabular malignancy. The purpose of this study was to evaluate the efficacy and safety of this combination. METHODS: Between October 2011 and May 2021, 11 patients with malignancies involving the acetabulum received hemipelvic replacement with a 3D-printed prosthesis and dual mobility bearing. Follow-up of postoperative survival, complications, and Musculoskeletal Tumor Society 93 (MSTS-93) lower limb functional scores were carried out. A finite element model of the postoperative pelvis was developed and input into the finite element analysis software. The Von Mises equivalent stress formula was used to analyze the stress distribution of each part of the pelvis under one gait cycle and the stress distribution at different angles of the hip joint. RESULTS: By the last follow-up, 9 of the 11 patients (81.8%) were still alive, and 2 patients had local tumor recurrence. The complications including 1 deep infection and 1 dislocation of the artificial joint. Excluding 1 amputation patient, the average score of the remaining 8 patients at the last follow-up was 21.4/30 (71.3%) on the MSTS-93. In the reconstructed pelvis, stress distributions were concentrated on the junction between hemipelvic prosthesis and screw and iliac bone on the resected side, and between femoral prosthesis stem and femoral bulb, while the stress of polyethylene lining was small. Before impact, the polyethylene lining will rotate at a small angle, about 3°. The inner stress of polyethylene liner is greater than the outer stress in all conditions. The polyethylene liner has no tendency to slide out. CONCLUSION: Pelvic tumor resection and reconstruction using 3D-printed hemipelvic prosthesis combined with dual mobility bearing was an effective treatment for pelvic tumors. Our patients achieved good early postoperative efficacy and functional recovery. The dual mobility bearing is beneficial to prevent dislocation, and the mechanical distribution and wear of the prosthesis are acceptable.

Bone Marrow Mesenchymal Stem Cell Exosome Attenuates Inflammasome-Related Pyroptosis via Delivering circ_003564 to Improve the Recovery of Spinal Cord Injury.

Bone marrow mesenchymal stem cell (BMSC) is previously reported to present a certain effect on treating spinal cord injury (SCI), while the underlying mechanism is largely uncovered. Therefore, the current study aimed to investigate the involvement of exosome-delivered circRNA profile in the BMSC's effect on pyroptosis for SCI treatment. H2O2 treated rat primary neurons were cultured with normal medium, BMSC, BMSC plus GW4869, and BMSC-derived exosome, respectively, then inflammasome-related pyroptosis markers, and circRNA profiles were detected. Subsequently, circ_003564-knockdown BMSC exosome was transfected into H2O2 treated rat primary neurons and NGF-stimulated PC-12 cells. Furthermore, in vivo validation was conducted. BMSC and BMSC-derived exosome both decreased inflammasome-related pyroptosis markers including cleaved caspase-1, GSDMD, NLRP3, IL-1ß, and IL-18 in H2O2-treated neurons, while exosome-free BMSC (BMSC plus GW4869) did not obviously reduce these factors. Microarray assay revealed that BMSC (vs. exosome-free BMSC) and BMSC-derived exosome (vs. normal medium) greatly regulated circRNA profiles, which were enriched in neuroinflammation pathways (such as neurotrophin, apoptosis, and TNF). Among three functional candidate circRNAs (circ_015525, circ_008876, and circ_003564), circ_003564 was most effective to regulate inflammasome-related pyroptosis. Interestingly, circ_003564-knockdown BMSC exosome showed higher expression of inflammasome-related pyroptosis markers compared to negative-control-knockdown BMSC exosome in H2O2 treated primary neurons/NGF-stimulated PC-12 cells. In vivo, BMSC exosome improved the function recovery and decreased tissue injury and inflammasome-related pyroptosis in SCI rats, whose effect was attenuated by circ_003564 knockdown transfection. BMSC exosome attenuates inflammasome-related pyroptosis via delivering circ_003564, contributing to its treatment efficacy for SCI.

Peptidomics Analysis Reveals Serum Biomarkers in Spinal Cord Injury Patients.

Spinal cord injury (SCI) is a severe traumatic disease, always resulting in neuronal injury. In this study, we aimed to exhibit a peptidome profile of serum from patients with SCI. A label-free peptidomics strategy was used to analyze the differentially expressed peptides (DEPs). Then, gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses was used to evaluate the function of the peptides precursors proteins. Also, the protein-protein interaction networks were mapped using STRING database. Finally, parallel reaction monitoring assays were used to validate the expression of candidate peptides. We identified 217 DEPs including 29 upregulated peptides and 188 downregulated peptides in SCI group. Many pathways such as Platelet activation, Complement and coagulation cascades, Focal adhesion were enriched. Seven peptides including PSPRPSP, RPPGFSP, DKPDMAEIEKFDKSKLK, STTAVVTNPKE, GHAGAQGPPGPPG, SMPPAQQQITS and SKVLPIQDNVSK were significantly changed between SCI patients and healthy people. Peptidomics provide a powerful tool to find the variation of SCI. RPPGFSP, DKPDMAEIEKFDKSKLK and SMPPAQQQITS may play important roles in SCI. However, the specific function of these peptides and whether they can be used as therapeutic targets for SCI need to be further investigated.

Immune Checkpoint Inhibitor Therapy for Bone Metastases: Specific Microenvironment and Current Situation.

The treatment of bone metastases is a thorny issue. Immunotherapy may be one of the few hopes for patients with unresectable bone metastases. Immune checkpoint inhibitors are the most commonly used immunotherapy drugs currently. In this review, the characteristics and interaction of bone metastases and their immune microenvironment were systematically discussed, and the relevant research progress of the immunological mechanism of tumor bone metastasis was reviewed. On this basis, we expounded the clinical application of immune checkpoint inhibitors for bone metastasis of common tumors, including non-small-cell lung cancer, renal cell carcinoma, prostate cancer, melanoma, and breast cancer. Then, the deficiencies and limitations in current researches were summarized. In-depth basic research on bone metastases and optimization of clinical treatment is needed.

Long intergenic non-coding RNA 511 correlates with improved prognosis, and hinders osteosarcoma progression both in vitro and in vivo.

BACKGROUND: This study aimed to investigate the correlation of long intergenic non-coding RNA 511 (LINC00511) with clinicopathological characteristics and overall survival (OS) in osteosarcoma patients and to explore its function in osteosarcoma in vitro and in vivo. METHODS: Tumor tissues and adjacent tissues from 45 osteosarcoma patients were acquired, and LINC00511 expression was detected. In vitro, LINC00511 expression was detected in osteosarcoma cell lines and osteoblast cell line. LINC00511 overexpression-treated (OE-LINC00511) and nonsense overexpression-treated (OE-control) MG-63 and Saos-2 cells were cultured, followed by the assessment of cell proliferation, apoptosis, migration, and invasion. In vivo, tumor weight and volume were measured in OE-LINC00511 and OE-control xenografted mice. RESULTS: LINC00511 expression was decreased in tumor tissues compared with adjacent tissues (P < .001), and its high expression correlated with increased tumor cell necrosis rate to neoadjuvant chemotherapy (P = .025) and prolonged OS (P = .010). In vitro, LINC00511 expression was decreased in osteosarcoma cell lines (including MG-63, U-2OS, Saos-2, and HOS) compared with osteoblast cell line (All P < .001). Cell proliferation was decreased at 48 hours (Both P < .01) and 72 hours (Both P < .001) (in MG-63 and Saos-2 cells); cell apoptosis was increased (P < .05) (in Saos-2 cells); cell migration and invasion were decreased (All P < .01) (in MG-63 cells and Saos-2 cells) in OE-LINC00511 compared with OE-control. In vivo, tumor volume was reduced at week 4 (P < .001), week 5 (P < .001), week 6 (P < .001) in OE-LINC00511 compared with OE-control. Tumor weight was declined in OE-LINC00511 than OE-control (P < .001). CONCLUSIONS: LINC00511 acts as a potential biomarker and therapeutic option for osteosarcoma.