Hongying Cao

Immunological memory triggered recurrence is the critical challenge in treating psoriasis. Tissue resident memory T (Trm) cells is the primary pacemaker in recurrent psoriasiform dermatitis, but the mechanism of activating Trm cells is still unclear. In this study, imiquimod induced recurrent psoriatic mice were established to reveal the phenotype of different Trm cells. Then, CD8⁺/CD4⁺ Tcm cells injection and NF-κB inhibitor/agonist were used to investigate the role and mechanism of Trm in recurrent psoriasis. Mice displayed severe dermatitis, high PASI scores and inflammatory cytokines after repeated imiquimod treatment. The CD8⁺ Trm cells ,but not CD4⁺ Trm cells, were elevated in skin of recurrent psoriatic mice. The CD8⁺ Tcm cells injection elicited more severe psoriatic symptom than imiquimod treatment alone, which indicate CD8⁺ Trm cells is critical participant in psoriatic recurrence. Meanwhile, the phosphorylated of NF-κB p52 and NF-κB p65 (RELA) both were enhanced in skin of imiquimod induced recurrent psoriatic mice. On the contrary, NF-κB inhibitor/agonist significantly suppressed/restore the dermatitis and CD8⁺ Trm cells in recurrent psoriatic mice. Hence, canonical and non-canonical NF-κB signaling both may contribute to activate CD8⁺ Trm cells in recurrent psoriasis.

Background: Diabetic cystopathy (DCP) is linked to bladder nerve conduction disorders, with diabetes-induced neuropathy impairing nerve signal transmission and causing bladder dysfunction. Myosin 5a, vital for neuronal transport, has been linked to neurological disorders, though its role in DCP remains unclear. The objective of this study was to investigate whether Myosin 5a plays a potential regulatory role in Diabetic Cystopathy. Methods: Bladder strips from diabetic rats were use to assess heightened responsiveness to external stimuli. Urodynamic assessments were conducted to track the progression of bladder voiding dysfunction over time, following streptozotocin (STZ) injection. Single-cell RNA-Seq mining was employed to identify associations between Myosin 5a and bladder overactivity. Cellular and tissue analyses were performed to determine the co-localization of Myosin 5a with neurotransmitter-related proteins. The impact of Myosin 5a knockdown on ChAT and SP expression in bladder neurons was also evaluated. Additionally, Myosin 5a-deficient DBA mice were studied for voiding function and sensitivity to stimuli. Student's t-test (two-tailed) or Mann–Whitney’s U test analysis of variance was used to analyze the difference between groups. Results: Bladder strips from diabetic rats exhibit increased responsiveness to external stimuli, with urodynamic assessments showing a progressive decline in bladder function, culminating in overactivity by the fourth week post-STZ injection. Co-localization of Myosin 5a with neurotransmitter-related proteins was observed, and the knockdown of Myosin 5a in bladder neurons led to a significant reduction in ChAT and SP expression. Myosin 5a-deficient DBA mice exhibited abnormal voiding function and reduced sensitivity to stimuli, along with significant downregulation of SLC17A9. Single-cell RNA-Seq analysis revealed a significant link between Myosin 5a and bladder overactivity, with Myosin 5a expression escalating in tandem with the severity of bladder dysfunction. Conclusions: Myosin 5a's dysregulation in diabetic rats may worsen bladder overactivity, suggesting its potential as a therapeutic target for diabetic OAB.

Overactive bladder (OAB) is a common, long-term symptom complex with a high prevalence in women worldwide. OAB has caused a social burden, and effective treatments are urgently needed. However, the pathogenesis of OAB has yet to be elucidated. Model rats underwent bladder outlet obstruction surgery. In the 2nd, 3rd, and 4th weeks after surgery, metabolic cages were used to detect the 12 h urine volume of rats in the sham and model groups. The urodynamic parameters bladder leak point pressure (BPLL), maximum voiding pressure (MVP), residual volume (RV), maximum bladder capacity (MBC), bladder compliance (BC), voided efficiency (VE), and non-voiding contractions (NVCs) were also detected. Moreover, the contractile responses of isolated detrusor muscles to electrical and carbachol stimulation were examined at the abovementioned time points. At the 4th week after surgery, the bladders of both groups were obtained for hematoxylin–eosin (H&E) and Masson’s trichrome staining. Real-time qPCR and Western blot were performed to quantify the expression of choline acetyltransferase (ChAT) and solute carrier family 17 member 9 (SLC17A9). At week 4, compared with the sham group, the 12 h urine volume of PBOO group increased significantly. The BLPP, MVP, VE, MBC, and NVCs increased significantly, and the VE was significantly reduced in 4-week PBOO group. The contractile responses of isolated detrusor muscles to electrical and carbachol stimulation significantly increased in 4-week PBOO group. In the 4-week PBOO group, the bladder wall and the ratio of bladder muscle to collagen within the bladder smooth muscle layer wall were significantly higher than those in the sham group. ChAT and SLC17A9 mRNA and protein expression in the OAB model rats significantly increased. At 4 weeks after PBOO, the OAB model was successfully established. The gene and protein expression levels of ChAT and SLC17A9 increased in the bladder of the OAB model, suggesting that OAB may be related to increased excitatory purinergic and cholinergic expression.

Persistent Helicobacter pylori infection causes a variety of gastrointestinal diseases and even gastric cancer. H. pylori invades gastric epithelial cells to survive and proliferate, which is one of the key factors in persistent colonization. A Published study has confirmed that cells can eliminate intracellular H. pylori through xenophagy to maintain intracellular balance. However, a growing body of evidences indicate that H. pylori can inhibit xenophagy by miRNA through regulating the expression of key autophagy-related genes. Through western blot analysis, mRFP-GFP-LC3 transfection assay, and transmission electron microscopy, we found that H. pylori infection obstructed autophagy flux degradation stage in GES-1 cell lines. Gentamicin protection assay confirmed that inhibit xenophagy is benefit for intracellular H. pylori survive. miR-30c-1-3p and miR-30c-5p were upregulated in GES-1 cell lines after infecting with H. pylori, resulting in the negative regulation on xenophagy. Further studies through bioinformatics analysis and dual-luciferase reporter assays confirmed that ATG14 and ULK1 were the target genes of miR-30c-1-3p and that ATG12 was the target gene of miR-30c-5p. The overexpression of miR-30c-1-3p and miR-30c-5p reduces the expression of ATG14, ULK1, and ATG12 at mRNA level and also decreased intracellular H. pylori elimination in GES-1 cells. The above results suggested that the inhibition on xenophagy by miR-30c-1-3p and miR-30c-5p through ATG14, ULK1, and ATG12 targeting benefitted intracellular H. pylori in the evasion of xenophagy clearance.

Aim: This research aimed to investigate the neurotoxicity of low-dose cyclophosphamide (CYP) on the urinary bladder of rats by in vivo and in vitro studies.Methods: To establish CYP-induced cystitis rat model, rats were treated with three intraperitoneal injections of CYP (25 mg/kg) in a week. During treatment, the up-down method was used to assess the mechanical withdrawal threshold. On day 8, urodynamic test and bladder smooth muscle contractility study, including the contraction of bladder strips to electrical field stimulation (EFS, 2–64 Hz), carbachol (CCh, 10^–8–10^–5 M) and KCl (120 mM), were performed to evaluate the function of bladder function. Body weight and bladder weight were also recorded. Morphometric analysis using an optical microscope and transmission electron microscope was performed to observe the changes of microstructure and submicrostructure of the bladder. The major pelvic neurons were isolated and treated with acrolein (the main CYP metabolite) to assess apoptosis in vitro. RT-PCR assays were used to quantify the mRNA expression levels of Nlrp6, Asc, Casp11 and Casp1 in bladder tissues and primary neurons.Results: After CYP injections, the body weights decreased, but the bladder weights increased in the model group. The mechanical withdrawal threshold of the cystitis model remained at a low level. The morphometric analysis suggested bladder inflammation and neuroinflammation in the bladder of the cystitis rat model. Urodynamic test revealed that, the amplitude, the pressure baseline, the peak pressure and pressure threshold of model rats significantly increased after CYP treatment. The muscle strips of model rats exhibited significantly higher contractility caused by EFS and CCh than the controls. Apoptotic cells appeared at the highest concentration group (100 μM acrolein) after 6 h of acrolein incubation in apoptosis assay of primary neurons. The mRNA expression levels of Nlrp6 and Casp11 were significantly increased in the cystitis rat model and in the acrolein-treated neurons.Conclusions: Low-dose CYP treatment was confirmed to induce nerve injury, which leading to bladder pain and overactive bladder in female rats, and the up-regulation of Nlrp6 and Casp11 may contribute to these pathological changes.