The management of antibiotic-resistant, bacterial biofilm infections in skin wounds poses an increasingly challenging clinical scenario. Pseudomonas aeruginosa infection is difficult to eradicate because of biofilm formation and antibiotic resistance. In this study, we identified a new benzothiazole derivative compound, SN12 (IC₅₀ = 43.3 nmol/L), demonstrating remarkable biofilm inhibition at nanomolar concentrations in vitro. In further activity assays and mechanistic studies, we formulated an unconventional strategy for combating P. aeruginosa-derived infections by targeting the two-component (Gac/Rsm) system. Furthermore, SN12 slowed the development of ciprofloxacin and tobramycin resistance. By using murine skin wound infection models, we observed that SN12 significantly augmented the antibacterial effects of three widely used antibiotics-tobramycin (100-fold), vancomycin (200-fold), and ciprofloxacin (1000-fold)-compared with single-dose antibiotic treatments for P. aeruginosa infection in vivo. The findings of this study suggest the potential of SN12 as a promising antibacterial synergist, highlighting the effectiveness of targeting the two-component system in treating challenging bacterial biofilm infections in humans.

Autophagy is a highly conserved mechanism for material degradation and recycling in eukaryote cells, and plays important roles in growth, development, stress tolerance and immune responses. ATG10 plays a key role in autophagosome formation. To understand the function of ATG10 in soybean, two homologous GmATG10 genes, namely GmATG10a and GmATG10b, were silenced simultaneously by bean pod mottle virus (BPMV) induced gene silencing. The carbon starvation induced by dark treatment and Western blotting analysis of GmATG8 accumulation level indicated that concurrent silencing GmATG10a/10b resulted in the impairment of autophagy in soybean; disease resistance and kinase assays demonstrated that GmATG10a/10b participated in the immune responses by negatively regulating the activation of GmMPK3/6, indicating that GmATG10a/10b plays a negative regulatory role in immune response in soybean.
Soybeans/genetics* ; Immunity

Objective:To investigate the role of IL-6 trans-signaling pathway in perioperative neurocognitive disorder in mice.Methods:Eighty-four SPF healthy male C57BL/6 wild-type mice and 84 SPF healthy male IL-6R -/- mice, aged 12-14 weeks, weighing 25-35 g, were used.The 84 wild-type mice were divided into 4 groups ( n=21 each) using a random number table method: sham group (SH group), surgery group (S group), sgp130Fc (specific IL-6 trans-signaling pathway blocker) group (F group), and sgp130Fc+ surgery group (FS group). In S group and FS group, internal fixation was performed under general anesthesia with sevoflurane after tibial fracture.Mice only received anaesthesia with sevoflurane in SH group and F group.In FS group and F group, sgp130Fc 10 mg/kg was intraperitoneally injected before anesthesia.Blood samples were collected from the celiac vein at 24 h after surgery for determination of the concentrations of interleukin 6 (IL-6), IL-1β and tumor necrosis factor (TNF)-α in plasma by enzyme-linked immunosorbent assay (ELISA). Then the mice were sacrificed, brains were removed, and hippocampal tissues were obtained for measurement of the contents of IL-6, IL-1β and TNF-α (by ELISA) and for observation of activation of microglias in the hippocampal DG region (by immunofluorescence staining, n=6). Cognitive function was evaluated by contextual fear conditioning test ( n=15) on 3 days after surgery.Eighty-four IL-6R -/- mice were randomly divided into 4 groups ( n=21 each): sham group (KO-SH group), surgery group (KO-S group), saline group (KO-C group), and hyper IL-6 (specific IL-6 trans-signaling pathway activator) group (KO-H group). The treatment in KO-SH group and KO-S group was the same as those previously described in SH group and S group, respectively.0.9% NaCl solution 100 μl was intraperitoneally injected in KO-C group, 100 μl hyper IL-6 40 μg/kg was intraperitoneally injected in KO-H group, and 24 h later blood was collected from the celiac vein for measurement of the concentrations of IL-6, IL-1β and TNF-α in plasma by ELISA.Then the mice were sacrificed, brains were removed, and hippocampal tissues were obtained for determination of the contents of IL-6, IL-1β and TNF-α (by ELISA) and for observation of activation of microglias in the hippocampal DG region (by immunofluorescence staining, n=6). Cognitive function was evaluated by contextual fear conditioning test ( n=15) on 3 days after surgery. Results:Compared with SH group, the percentage of freezing time in the contextual fear conditioning test was significantly decreased, and the activation of microglias in the hippocampal DG region and levels of IL-6, IL-1β and TNF-α in plasma and hippocampi were increased in S group ( P<0.05). Compared with S group, the percentage of freezing time in the contextual fear conditioning test was significantly increased, and the activation of microglias in the hippocampal DG region and levels of IL-6, IL-1β and TNF-α in plasma and hippocampus were decreased in FS group ( P<0.05). There were no significant differences in the percentage of freezing time, activation of microglias in the hippocampal DG region, and levels of IL-6, IL-1β and TNF-α in plasma and hippocampi between KO-S group and KO-SH group ( P>0.05). Compared with KO-C group, the percentage of freezing time in the contextual fear conditioning test was significantly decreased, and the activiation of microglias in the hippocampal DG region and levels of IL-6, IL-1β and TNF-α in plasma and hippocampus were increased in KO-H group ( P<0.05). Conclusions:IL-6 trans-signaling pathway is involved in the process of perioperative neurocognitive disorder in mice.

Objective:To evaluate the test-retest reliability of MRI criteria in the 2019 Bosniak classification of cystic renal masses (CRMs) and to analyze the impact of lesions′ property, size and readers′ experience on the test-retest reliability.Methods:From January 2009 to June 2019, 207 patients with 207 CRMs were included in this retrospective study. All of them underwent renal MRI and surgical-pathologic examination. According to Bosniak classification, version 2019, all CRMs were independently classified twice by eight radiologists with different levels of experience. All radiologists were blinded to the pathology of the lesions. By using intraclass correlation coefficient (ICC), test-retest reliability was evaluated for all CRMs and for subgroups with different pathological properties (benign and malignant) and different sizes (≤40 mm and>40 mm). The test-retest reliability of 4 senior readers (≥10 years of experience) and 4 junior readers (<10 years of experience) were evaluated respectively. The comparison of ICC was performed using Z test. Results:The 207 CRMs included 111 benign lesions (83 benign cysts, 28 benign tumors) and 96 malignant tumors. There were 87 lesions with maximum diameter ≤40 mm and 120 with maximum diameter>40 mm. The test-retest reliability (ICC) of each reader for all lesions was 0.776-0.888, the overall ICC was 0.848 (95%CI 0.821-0.872). The ICCs of senior and junior readers were 0.853 (95%CI 0.824-0.880) and 0.843 (95%CI 0.811-0.871) respectively, without significant difference between the two groups ( Z=0.85, P=0.374). The ICC of all readers was 0.827 for benign lesions and 0.654 for malignant lesions, showing significant difference ( Z=2.80, P=0.005). The ICC was 0.770 for lesions ≤40 mm and 0.876 for lesions>40 mm, which was significantly different ( Z=-2.36, P=0.018). For CRM subgroups with different pathological properties and different sizes, there was no significant difference in test-retest reliability between senior and junior readers (all P>0.05). Conclusion:The test-retest reliability of MRI criteria in the 2019 Bosniak classification of CRMs is excellent and unaffected by readers′ experience. The reliabilities are not consistent among CRMs of different pathological properties and different sizes, but all reached the level of good and above.

Inflammatory caspase-11 senses and is activated by intracellular lipopolysaccharide (LPS) leading to pyroptosis that has critical role in defensing against bacterial infection, whereas its excess activation under pathogenic circumstances may cause various inflammatory diseases. However, there are few known drugs that can control caspase-11 activation. We report here that scutellarin, a flavonoid from Erigeron breviscapus, acted as an inhibitor for caspase-11 activation in macrophages. Scutellarin dose-dependently inhibited intracellular LPS-induced release of caspase-11p26 (indicative of caspase-11 activation) and generation of N-terminal fragment of gasdermin D (GSDMD-NT), leading to reduced pyroptosis. It also suppressed the activation of non-canonical nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome as evidenced by reduced apoptosis-associated speck-like protein containing a CARD (ASC) speck formation and decreased interleukin-1 beta (IL-1β) and caspase-1p10 secretion, whereas the NLRP3-specific inhibitor MCC950 only inhibited IL-1β and caspase-1p10 release and ASC speck formation but not pyroptosis. Scutellarin also suppressed LPS-induced caspase-11 activation and pyroptosis in RAW 264.7 cells lacking ASC expression. Moreover, scutellarin treatment increased Ser/Thr phosphorylation of caspase-11 at protein kinase A (PKA)-specific sites, and its inhibitory action on caspase-11 activation was largely abrogated by PKA inhibitor H89 or by adenylyl cyclase inhibitor MDL12330A. Collectively, our data indicate that scutellarin inhibited caspase-11 activation and pyroptosis in macrophages at least partly via regulating the PKA signaling pathway.

Dynamic variations of the cell microenvironment can affect cell differentiation, cell signaling pathways, individual growth, and disease. Optogenetics combines gene-encoded protein expression with optical controlling, and offers a novel, reversible, non-invasive and spatiotemporal-specific research tool to dynamically or reversibly regulate cell signaling pathways, subcellular localization and gene expression. This review summarizes the types of optogenetic components and the involved cellular signaling pathways, and explores the application and future prospects of the light-controlled cell signaling pathways.
Cell Differentiation ; Light ; Optogenetics ; Proteins ; Signal Transduction

Objectives To evaluate the role of PCSK9 (proprotein convertase subtilisin kexin type 9) on the lipid accumulation and kidney injury of C57BL/6 mice. Methods The 24 h urine of 12 weeks old wide type C57BL/6 mice and PCSK9 knockout (KO) mice were collected through a metabolic cage, followed by perfusion and sacrifice. Urinary microalbumin?to?creatinine ratio (UACr), total cholesterol and triglyceride in kidney tissues were measured by ELISA. BODIPY 493/503 staining and standard transmission electron microscopy (TEM) of kidney tissues was performed for evaluating lipid accumulation and podocyte foot effacement in the kidney. Kidney tissues were also evaluated by PAS stain and TUNNEL stain. PCSK9, podocin and nephrin were quantified through real?time PCR, and the Bcl?2, Bax and cleaved caspase 3 were evaluated by Western blotting. Results Total cholesterol and triglyceride contents were higher in the kidneys of PCSK9 KO mice than controls (P<0.05). The level of lipid accumulation in glomeruli and tubules through BODIPY 493/503 stain, and the amount of lipid drop in TEM were more serious in PCSK9 KO mice. UACr and podocyte foot process effacement were increased, and the transcription of podocin and nephrin were decreased in the kidneys of PCSK9 KO mice (all P<0.05). The expression of Bcl?2 was decreased, and Bax and cleavedcaspase 3 were increased in the kidney samples of PCSK9 KO mice. Conclusion PCSK9 might be reversely involved in lipid homeostasis and accumulation, resulting in injury and apoptosis in the kidneys of C57BL/6 mice.

Although still in its infant stage, synthetic biology has achieved remarkable development and progress during the past decade. Synthetic biology applies engineering principles to design and construct gene circuits uploaded into living cells or organisms to perform novel or improved functions, and it has been widely used in many fields. In this review, we describe the recent advances of mammalian synthetic biology for the treatment of diseases. We introduce common tools and design principles of synthetic gene circuits, and then we demonstrate open-loop gene circuits induced by different trigger molecules used in disease diagnosis and close-loop gene circuits used for biomedical applications. Finally, we discuss the perspectives and potential challenges of synthetic biology for clinical applications.