OBJECTIVETo develop a new method for the determination of the idiosyncratic component, 2,3,5,4'-tetrahydroxy-stilbene-2-O-beta-D-glycoside in Shengfa Powders.

METHODA HPLC method was set up, using Hypersil BDS C18 column (5 microns, 4.6 mm x 150 mm), acetonitrile-water(18:82) as mobile phase, with detection at 320 nm.

RESULTThe calibration curve was linear in the range of 0.45-2.25 micrograms, r = 0.9998, the average recovery was 99.5%, RSD = 1.2%(n = 6).

CONCLUSIONThe active constituent 2,3,5,4'-tetrahydroxy-stilbene-2-O-beta-D-glycoside in Shengfa Powders can be separated effectively. This method is simple, specific and exact.

Drug Combinations ; Drug Stability ; Drugs, Chinese Herbal ; analysis ; Glycosides ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Powders ; chemistry ; Quality Control ; Stilbenes ; analysis

OBJECTIVETo evaluate the protective efficacy of plague subunit vaccine, BALB/c mice, guinea pigs and rabbits were used in this study.

METHODSGroups of mice (10 per group), guinea pigs (14 per group) and rabbits (6 per group) were immunized with F1 + rV270 vaccine, EV76 vaccine and alum adjuvant by intramuscular route, respectively. Serum antibody titres of mice, guinea pigs and rabbits were determined by ELISA and the immunized animals were challenged with 10(6) CFU of Y. pestis strain 141 at the 8th week after the primary immunization.

RESULTSThe immunized mice, guinea pigs or rabbits with subunit vaccine developed anti-F1 IgG titre of 41 587.3 +/- 2.1, 11 543.7 +/- 2.1 or 522.4 +/- 22.4 and elicited statistical anti-F1 IgG titre difference among them (F = 17.58, P < 0.01). The immunized mice, guinea pigs or rabbits with subunit vaccine had anti-rV270 IgG titre of 15 748.7 +/- 1.6, 12.6 +/- 1.4 or 1648.0 +/- 5.0 and induced statistical anti-rV270 IgG titre difference among them (F value was 16.34, P < 0.01). There was significant anti-F1 IgG titre difference among mice, guinea pigs and rabbits immunized with EV76 vaccine that developed anti-F1 IgG titre of 913.4 +/- 4.5, 937.0 +/- 2.0 or 342.0 +/- 12.0 (F = 23.67, P < 0.01), whereas the immunized mice, guinea pigs and rabbits with EV76 vaccine developed anti-rV270 IgG titre of 12.0 +/- 1.0, 447.0 +/- 10.0, 40.0 +/- 11.0 and there was no anti-rV270 IgG titre difference between them (F = 2.20, P = 0.1314). The immunized mice with subunit vaccine developed significantly higher anti-F1 IgG titres than immunized guinea pigs and rabbits (q value was 30.57 and 19.04, respectively, P < 0.01), and there were no anti-F1 IgG titre differences between the immunized guinea pigs and rabbits (q = 0.04, P = 0.8485). The immunized mice with subunit vaccine developed significantly higher anti-rV270 IgG titres than immunized guinea pigs and rabbits (q value was 27.10 and 19.49, respectively, P < 0.01), and there were no anti-rV270 IgG titre differences between the immunized guinea pigs and rabbits with the subunit vaccine (q = 0.25, P = 0.6187). The immunized mice with EV76 elicited higher anti-F1 IgG titres than immunized guinea pigs and rabbits (q value was 40.67 and 29.10, respectively, P < 0.01), whereas there was no difference of F1 IgG titer between immunized guinea pigs and rabbits (q = 0.06, P = 0.8098). The immunized mice, guinea pigs and rabbits with subunit vaccine provided 100% (10/10), 86% (12/14) and 100% (5/5) protection against 10(6) CFU Y. pestis of challenge, respectively. The immunized mice, guinea pigs and rabbits with EV76 vaccine gave 100% (6/6), 93% (13/14) and 100% (6/6) protection against 10(6) CFU Y. pestis of challenge respectively.

CONCLUSIONBALB/c mice is the best small animal model for valuation of protective efficacy of plague subunit vaccine. The guinea pigs showed a high individual variation for this purpose. The rabbits can be used as an alternative model for evaluating plague subunit vaccine.

Animals ; Antibodies, Bacterial ; blood ; Dose-Response Relationship, Immunologic ; Female ; Guinea Pigs ; Immunization ; Immunoglobulin G ; blood ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Plague ; prevention & control ; Plague Vaccine ; immunology ; Rabbits ; Vaccines, Subunit ; immunology

【Objective】To explore the clinical manifestation of COVID- 19 severe cases.【Methods】Clinical data of one severe case with COVID-19 including the clinical characteristic ，laboratory testing results，radiography，treatment，complication and outcome of the patient were retrospectively collected and analyzed.【Results】 The patient with COVID-19 was a 61-year old male，He suffered with underlying disease. His symptoms included fever，cough，myalgia， fatigue，and dyspnea. Laboratory testing results included normal WBC count，decreased lymphocyte cells，elevated LDH and hypoxemia. Radiography findings showed bilateral lung infiltration. His condition deteriorated after intensive treatment for one week. He was intubated and treated with mechanical ventilation because of complicating with severe acute respiratory distress syndrome（ARDS）.【Conclusion】COVID-19 is an emerging acute communicable disease，which lack specific and effective treatment. Most patients have a good prognosis but mortality in severe cases is high. More attention should be paid on the high risk of progression in COVID-19 cases.

【Objective】 To explore the role of NLRP3 inflammasome in methicillin-resistant staphylococcus aureus(MRSA) infection secondary to influenza A virus H1N1(IAV H1N1) in vitro. 【Methods】 Macrophages RAW264.7 were cultured and then infected with only MRSA for 24 h(MRSA group) and with MRSA for 24 h secondary to H1N1 infection for 1 week in advance(MRSA+ H1N1 group), respectively. Fluorescence quantitative PCR was applied to detect the transcription of NLRP3, Caspase-1 and IL-1β, immunofluorescence and western blot were used to detect NLRP3 protein in cells, and the concentration of IL-1β in supernatant was measured by enzyme linked immunosorbent assay(ELISA) . 【Results】 In MRSA group, the transcriptions of NLRP3 and Caspase-1 mRNA, as well as translation of NLRP3, showed no difference compared with control group, while the expression of IL-1β mRNA and the concentration of IL-1β in supernatant were significantly higher than those in control group(both P<0.01). In H1N1+MRSA group, the transcription of NLRP3 and Caspase-1 were significantly higher than those in control group(both P<0.01), the translation of NLRP3 was significantly higher than that in control group(P<0.01), the concentration of IL-1β was significantly higher than that in control group(P<0.01). In H1N1+MRSA group, the transcription and translation of NLRP3 were significantly higher than those in MRSA group(both P<0.01), while the transcription of IL-1β was lower than that in MRSA group, and the concentration of IL-1β in supernatant was significantly lower than that in MRSA group(P<0.01) . 【Conclusions】 Our study suggests that IL-1β secretion induced by MRSA infection is in a NLRP3 inflammasome independent manner in macrophage. It also suggests that influenza A virus H1N1 infection in advance decreases the release of IL-1β induced by secondary MRSA infection ultimately, which may contribute to the mechanism of MRSA pneumonia secondary to IAV infection.