This study aimed to evaluate the effect of sanguinarine on biomechanical properties of rat airway smooth muscle cells (rASMCs) including stiffness, traction force and cytoskeletal stress fiber organization. To do so, rASMCs cultured in vitro were treated with sanguinarine solution at different concentrations (0.005~5 μmol/L) for 12 h, 24 h, 36 h, and 48 h, respectively. Subsequently, the cells were tested for their viability, stiffness, traction force, migration and microfilament distribution by using methylthiazolyldiphenyl-tetrazolium bromide assay, optical magnetic twisting cytometry, Fourier transform traction microscopy, scratch wound healing method, and immunofluorescence microscopy, respectively. The results showed that at concentration below 0.5 μmol/L sanguinarine had no effect on cell viability, but caused dose and time dependent effect on cell biomechanics. Specifically, rASMCs treated with sanguinarine at 0.05 μmol/L and 0.5 μmol/L for 12 and 24 h exhibited significant reduction in stiffness, traction force and migration speed, together with disorganization of the cytoskeletal stress fibers. Considering the essential role of airway smooth muscle cells (ASMCs) biomechanics in the airway hyperresponsiveness (AHR) of asthma, these findings suggest that sanguinarine may ameliorate AHR via alteration of ASMCs biomechanical properties, thus providing a novel approach for asthma drug development.
ObjectiveTo explore the clinical value of next-generation sequencing (NGS) in the diagnosis of Pneumocystis jirovecii pneumonia (PCP).MethodsTwo patients with Pneumocystis jirovecii pneumonia after lung transplantation were detected by NGS in the sputum and bronchoalveolar lavage fluid. The clinical data, imaging features, laboratory examination and treatment of the two patients were reported. A systematic literature review was performed for similar published cases in PubMed database, using the keywords "lung transplantation/solid organ transplantation" and "Pneumocystis jirovecii".ResultsThere were six references based on the keywords of "lung transplantation " and " Pneumocystis jirovecii ", of which three were case report. Sixty-six lung transplant patients were complicated with Pneumocystis jirovecii in total. The clinical manifestations of Pneumocystis jirovecii pneumonia were fever and dyspnea of different degrees. The diffuse "ground glass" infiltration could be shown on imaging. Computer tomography scan of chest was a sensitive method to detect PCP. Combined immunofluorescence microscope/PCR/serum 3-β-D-glucan could effectively improve the accuracy of microbiology detection. In addition, NGS could quickly and accurately identify pathogenic bacteria, give guidance for treatment and improve prognosis so as to benefit patients well. Trimethoprim/sulfamethoxazole (TMP/SMZ) was the preferred choice for the treatment of PCP patients.ConclusionsPneumocystis jirovecii pneumonia is more common in patients with immunodeficiency or immunosuppression. NGS can help rapid and accurate diagnosis, and the treatment should be early and sufficient.