Objective To investigate the expression of inducible nitric oxide synthase ( iNOS) and the levels of nitric oxide (NO) in THP-1 and J774A. 1 cells during Leptospira interrogans (L. interrogans) infection for a better understanding of the mechanism of macrophages involved defense against L. interrogans strains in different hosts. Methods The human mononuclear macrophages (THP-1) and the murine mono-nuclear macrophages (J774A. 1) were infected with L. interrogans strain 56601. The expression of iNOS at mRNA and protein levels were determined by using real-time RT-PCR and flow cytometry analysis. The lev-els of NO were detected with Griess test. Results The expression of iNOS at mRNA level in J774A. 1 and THP-1 cells infected with L. interrogans strains for 2, 4, 12 and 24 hours were respectively 1. 37, 2. 82, 25. 76, 27. 47 times and 1. 59, 3. 98, 3. 89, 8. 81 times than that of cells without infection (P<0. 05). The expression rates of iNOS protein in J774A. 1 cells were increased from 34. 16% to 85. 85%, 93. 82%, 91. 77% and 93. 65% along with the increased time of infection time (P<0. 05). The expression rates of iNOS protein in THP-1 cells were up-regulated from 22. 08% to 72. 64%, 81. 33%, 80. 03% and 65. 72%after 2, 4, 12 and 24 hours of infection (P<0. 05), respectively. Results of the Griess test indicated that the levels of NO in J774A. 1 and THP-1 cells were respectively increased from 0. 1588 μmol/L to 0. 2208μmol/L, 0. 2668μmol/L, 0. 3808μmol/L, 0. 3828μmol/L and from 0. 0988μmol/L to 0. 2848μmol/L,0. 3228 μmol/L, 0. 2608μmol/L and 0. 3308μmol/L after infection with L. interrogans strains for 2, 4, 12 and 24 hours (P<0. 05). Conclusion The expression of iNOS and the levels of NO in J774A. 1 and THP-1 cells were significantly increased during L. interrogans infection. This study might help to explain the bactericidal mechanism of macrophages derived from different hosts against L. interrogans infection.

Objective To analyze the effects of Leptospira interrogans ( L. interrogans) infection on the activation of NLRP3 in THP-1 and J774A. 1 cells and to further understand the mechanism of inflam-mation caused by L. interrogans in different hosts. Methods Human mononuclear macrophage cell line (THP-1) and murine mononuclear macrophage cell line (J774A. 1) were infected with L. interrogans strain 56601. The expression of NLRP3 at mRNA and protein levels were measured by using real-time RT-PCR and flow cytometry analysis, respectively. The NLRP3-mediated secretion of IL-1β, IL-18 and IL-33 was detec-ted by ELISA combined with the NLRP3 inhibitory test. Results Compared with the normal cells, the ex-pression of NLRP3 at mRNA level in L. interrogans-infected THP-1 cells was respectively increased by 4. 05, 0. 34, 0. 33, 0. 06 and 1. 66 times at the time points of 1 h, 2 h, 4 h, 12 h and 24 h after infection ( P<0. 05), while that in L. interrogans-infected J774A. 1 cells was respectively increased by 12. 98, 16. 19, 10. 68, 5. 8 and 0. 57 times (P<0. 05). The expression rates of NLRP3 protein in THP-1 and J774A. 1 cells respectively increased from 9. 26% to 94. 01%, 89. 24%, 31. 80%, 19. 74%, 11. 28% and from 18. 71%to 58. 78%, 43. 64%, 36. 42%, 76. 46%, 85. 21% at the time points of 1 h, 2 h, 4 h, 12 h and 24 h af-ter L. interrogans infection (P<0. 05). The level of IL-1β in L. interrogans-infected THP-1 cells was 73. 07 pg/ml, 939. 24 pg/ml, 939. 24 pg/ml, 843. 22 pg/ml and 851. 06 pg/ml at the time points of 1 h, 2 h, 4 h, 12 h and 24 h, respectively (P<0. 05), while the level of IL-1β in L. interrogans-infected J774A. 1 cells began to rise at the time point of 12 h from 191. 17 pg/ml to 254. 4 pg/mL at the time point of 24 h (P<0. 05). The level of IL-18 in L. interrogans-infected THP-1 cells was 913. 89 pg/ml, 808. 19 pg/ml, 483. 54 pg/ml, 204. 19 pg/ml and 189. 09 pg/ml at the time points of 1 h, 2 h, 4 h, 12 h and 24 h, re-spectively (P<0. 05), while the level of IL-18 in L. interrogans-infected J774A. 1 cells increased at the time point of 24 h, which was 113. 37 pg/ml (P<0. 05). A slight increase in the level of IL-33 was detected in L. interrogans-infected J774A. 1 cells at the time points of 12 h and 24 h to 201. 14 pg/ml and 155. 68 pg/ml, respectively (P<0. 05), but no significant change was detected in L. interrogans-infected THP-1 cells (P>0. 05). Results of the inhibitory test showed that the up-regulation of IL-1β , IL-18 and IL-33 in THP-1 and J774A. 1 cells were effectively inhibited by the specific inhibitor of NLRP3. Conclusion NL-RP3 inflammasome was activated and involved in the production of specific inflammatory cytokines IL-1βand IL-18 in both THP-1 and J774A. 1 cells after L. interrogans infection, but the inflammatory cytokines induced by L. interrogans infection varied in different cells. L. interrogans induced earlier and higher level of IL-1βand IL-18 production in human macrophages than in murine macrophages.

Objective To discuss the contribution of clinical pharmacists in anti-infection treatment of a patient with multiple organ dysfunction syndrome ( MODS) undergoing continuous renal replacement therapy ( CRRT). Methods Pharmacists participated in the anti-infection treatment of a MODS patient undergoing CRRT.Pharmacists assisted physicians in optimizing the therapeutic regimen based on treatment guidelines and relative information. Results Physicians accepted the advice of pharmacists after comprehensive evaluation.Ten days later,the patient recovered from shock,as the infection indexes were improved significantly.He then moved back to the general ward. Conclusion Pharmacists should positively participate in clinical treatment with physicians,in order to play a critical role in ensuring the safety and efficacy of the medication.