Objective:To explore mechanism of Caspase-11/GSDMD non-classical cell pyroptosis pathway in inflammatory response of RAW264.7 monocyte macrophages induced by ricin toxin.Methods:Cell pyroptosis model induced by LPS+Nigericin was used as a positive control,and cells were induced with 40 ng/ml and 80 ng/ml of ricin toxin for 8 hours.Cell release of lactate dehydro-genase(LDH)was measured by LDH cell release assay;cell viability was assessed by flow cytometry(Annexin Ⅴ/PI double staining);gene expressions of Caspase-11,GSDMD,IL-1β and IL-18 were measured by RT-qPCR;protein levels of Caspase-11 and GSDMD were measured by Western blot;secretion levels of inflammatory cytokines IL-1β and IL-18 in cell supernatant were measured by ELISA.Results:Compared with normal control group,RAW264.7 cells treated with ricin toxin showed swelling,cell membrane rupture,significantly increased LDH release(P<0.001),significantly increased pyroptosis rate(P<0.001),significantly increased expression of pyroptosis-related genes Caspase-11,GSDMD,IL-1β and IL-18(P<0.05),significantly increased protein expressions of Caspase-11 and GSDMD(P<0.05),and significantly increased secretion levels of IL-1β and IL-18 in cell supernatant(P<0.05).Results of ricin toxin treatment group were consistent with LPS+Nigericin cell pyroptosis positive control group.Conclu-sion:Ricin toxin may induce cell pyroptosis in RAW264.7 cells through Caspase-11/GSDMD non-classical pathway,thereby promoting inflammatory response.

Objective:To explore mechanism of Caspase-11/GSDMD non-classical cell pyroptosis pathway in inflammatory response of RAW264.7 monocyte macrophages induced by ricin toxin.Methods:Cell pyroptosis model induced by LPS+Nigericin was used as a positive control,and cells were induced with 40 ng/ml and 80 ng/ml of ricin toxin for 8 hours.Cell release of lactate dehydro-genase(LDH)was measured by LDH cell release assay;cell viability was assessed by flow cytometry(Annexin Ⅴ/PI double staining);gene expressions of Caspase-11,GSDMD,IL-1β and IL-18 were measured by RT-qPCR;protein levels of Caspase-11 and GSDMD were measured by Western blot;secretion levels of inflammatory cytokines IL-1β and IL-18 in cell supernatant were measured by ELISA.Results:Compared with normal control group,RAW264.7 cells treated with ricin toxin showed swelling,cell membrane rupture,significantly increased LDH release(P<0.001),significantly increased pyroptosis rate(P<0.001),significantly increased expression of pyroptosis-related genes Caspase-11,GSDMD,IL-1β and IL-18(P<0.05),significantly increased protein expressions of Caspase-11 and GSDMD(P<0.05),and significantly increased secretion levels of IL-1β and IL-18 in cell supernatant(P<0.05).Results of ricin toxin treatment group were consistent with LPS+Nigericin cell pyroptosis positive control group.Conclu-sion:Ricin toxin may induce cell pyroptosis in RAW264.7 cells through Caspase-11/GSDMD non-classical pathway,thereby promoting inflammatory response.

Objective:To investigate the work requirement and training needs of general practitioners in Shenzhen during the outbreak of coronal virus disease 2019 (COVID-19).Methods:From February 19 to February 29, 2020, the questionnaires of "Coping with COVID-19 epidemic of genereal practitioners in Shenzhen region" were distributed to 4 002 general practitioners in Shenzhen hospitals and communities through the Questionnaire Star software. The changes of job content, education parttern, and the perception of roles before and during the epidemic were surveyed; the different roles of doctors engaged in general medicine before and after 2008 in education and community work were compared; the needs for training in epidemic prevention related knowledge for general practitioners were investigated.Results:A total of 382 questionnaires were recovered, of which 361 were valid, and the effective rate was 94.5%. The 83.1% (300/361) of general practitioners believed that the number of patients during the epidemic was significantly reduced. The main work was to screen community residents in epidemic areas (68.9%, 249/361), telephone counseling (54.0%, 195/361), and screening and follow-up of close contacts (42.9%, 155/361); 98.9% (357/361) of community doctors participated in outpatient education. Experienced doctors (starting work before 2008) were more willing to undertake management work in the epidemic (χ 2=77.370, P<0.05) . General practitioners′ needs for training were psychological interventions for residents under quarantine (80.6%, 291/361), home protection knowledge in the community (74.2%, 268/361), and isolation measures of the family with suspected patients (67.3%, 243/361), wearing and taking off protective gowns (65.1%, 235/361), collecting pharyngeal swab specimens (57.3%, 207/361) and patient referral procedures (52.4%, 189/361). The 60.6% (219/316) of general practitioners preferred information from the government press conference, expert opinion or the intranet of the organization (such as the hospital management department). Conclusion:General practitioners in Shenzhen took important screening, public education and counseling works during the COVID-19 epidemic, meanwile the training in the psychological intervention, the prevention and control knowledge related to the epidemic should be strengthend to better cope with the epidemics.