ObjectiveThe research provided the constructive ideas for evaluation and training of deans of tertiary hospital by analyzing the leadership status quo through using the situational judgment test based on the leadership contingency theory.MethodsBased on the leadership contingency theory,we designed situational judgment test for the dean of tertiary hospital.There were 215 participants involved in the leadership evaluation. Results The coaching leadership style occupied the mainly leadership style of deans of tertiary hospital.In the management situation matched by the coaching leadership style,the leadership of the deans was significantly higher than other management situations.Conclusion The main conclusion included,firstly,the coaching leadership style was the mainly leadership style of deans of tertiary hospital.It was strongly correlated with the deans＇ background thatthey were mainly from clinical professionals.Secondly,the leadership of the deans was at middle level;therefore,it needed to increase the capacity building of deans through management training.

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 investigate the effects of Leptospira interrogans ( L. interrogans) infec-tion on the activities of NADPH oxidase ( nicotinamide adenine dinucleotide phosphate-oxidase) and the lev-els of reactive oxygen species (ROS) in THP-1 and J774A. 1 cells and to understand the bactericidal mecha-nisms of macrophages in different hosts against L. interrogans. Methods Human mononuclear macrophage cell line (THP-1 cells) and murine mononuclear macrophage cell line (J774A. 1 cells) were infected with L. interrogans strain 56601. The activities of NADPH oxidase and the levels of superoxide ion ( O-2 ) were measured with spectrophotography. Changes in the levels of ROS were detected with immunofluorescence as-say. Results Compared with the normal cells, the activities of NADPH oxidase in L. interrogans-infected J774A. 1 cells changed from 0. 619 0 μmol · min-1 · mg-1 to 0. 305 5 μmol · min-1 · mg-1 , 6. 141 5μmol·min-1 ·mg-1 , 1. 487 1μmol·min-1 ·mg-1 and 0. 964 6μmol·min-1 ·mg-1 after 2, 4, 12 and 24 hours of infection, respectively (P<0. 05), while the activities of NADPH oxidase in L. interrogans-infected THP-1 cells were up-regulated from 0. 723 5μmol·min-1 ·mg-1 to 0. 884 2μmol·min-1 ·mg-1 , 1. 897 1μmol·min-1 ·mg-1 , 1. 125 4 μmol·min-1 ·mg-1 and 0. 562 7 μmol·min-1 ·mg-1 , respectively ( P<0. 05). The levels of O-2 in L. interrogans-infected J774A. 1 cells at the time points of 2 h, 4 h, 12 h and 24 h after infection increased from 0. 189 0μmol/L to 0. 236 3μmol/L, 0. 297 7μmol/L, 0. 324 0μmol/L and 0. 305 7 μmol/L, respectively (P<0. 05), while the levels of O-2 in L. interrogans-infected THP-1 cells rose from 0. 123 7 μmol/L to 0. 149 3 μmol/ L, 0. 249 0 μmol/ L, 0. 270 0 μmol/ L and 0. 272 7μmol/L, respectively (P<0. 05). The fluorescence intensity of ROS in THP-1 and J774A. 1 cells increased gradually after infection with L. interrogans for 2 h and decreased after reaching the peak at 24 h. Conclu-sion Both the activities of NADPH oxidase and the levels of O-2 in J774A. 1 and THP-1 cells were signifi-cantly upregulated after infected with L. interrogans, especially in J774A. 1 cells. The results of this study provided references for further elucidating the bactericidal mechanisms of macrophages in different hosts against L. interrogans.

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 conduct an epidemiological investigation of two leptospirosis death cases reported in Guizhou Province in 2014. Methods: The information of the patients were investigated and analyzed. The serological detection, samples of the two patients was detected using ELISA and microscopic agglutination test (MAT). Leptospira carrier status of murine host animal in the living environment of the two patients was investigated in October and November of 2014. Leptospires in the kidney were cultured and isolated, the isolates were identified using Leptospira specific PCR and further identified with serogroup specific PCR and the conventional MAT. The relativity between the carrier status of murine and the death cases of human leptospirosis was analyzed. Results: The two death cases of human leptospirosis came from Liping County and the clinical symptoms were consistent with the diagnosis criteria for Leptospirosis. The results of ELISA detection showed that the anti-Leptospira antibody was positive for one of the death cases, MAT identified the serum reacted with sera-group icterohaemorrhagiae Leptospira, while the serum sample of the other case was failed to perform antibody detection due to hemolysis. 1 600 traps were placed in the living environment of the two death cases and 183 murine rodents were trapped. The murine density was 11.44% (183/1 600); 40 leptospirea suspected strains were isolated and all of them were isolated from Apodemus agrarius. The positive rate was 21.86% (40/183); 95 Apodemus agrarius were trapped and the murine density was 5.93% (95/1 600). Species specific PCR identified all the 40 strains as Leptospire. Serogroup specific PCR further identification showed that they were iterohaemorrahgiae serogroup Leptospria. interrogans. Conclusion Anti-iterohaemorrahgiae Leptospira antibody was detected from one of the two patients. 40 strains of iterohaemorrahgiae serogroup Leptospira interrogans were isolated and all of them were isolated from Apodemus agrarius in the living environment and the serogroup of the Leptospira matched with the serological detection results from patients, which indicated that the two death cases were caused by the infection of iterohaemorrahgiae serogroup Leptospira interrogans, and Apodemus agrarius were the potential source of infection.

Objective:To understand the etiological characteristics of the patients with fever of unknown origin in Guizhou province through the isolation and identification of Leptospira interrogans and provide evidence for the control, prevention and treatment of human leptospirosis. Methods:Blood and urine samples were collected from patients with fever symptoms in Qiandongnan, an epidemic area, in Guizhou. The suspected Leptospira strains were primarily identified using pathogenic Leptospira specific G1/G2-PCR, and subsequently identified by using Leptospira serogroups specific PCR. The Leptospira strains were then genotyped with multiple locus sequence typing. MLST data based cluster analysis on the isolates and Leptospira reference strains of common serogroups were analyzed by using software NTsys 2.10e. Results:Three suspected strains of Leptospira were isolated from human blood samples, the isolation rate was 8.６%, which were designated as strain 17BX002, 17BX003 and 17AJX008. Strain 17BX002 was further identified as serogroup grippotyphosa by using Leptospira serogroup specific PCR, while the other two strains were negative (excluded as iterohaemorrhagiae, sejroe, canicola, autumnalis, grippotyphosa and hebdomadis). MLST genotyping showed that strain 17BX002 was typed as ST106, most closely clustered with Leptospira grippotyphosa, while strain 17BX003 and 17AJX008 were typed as ST96, the same as serogroup badaviae. Conclusion:There are leptospirosis cases in epidemic area of Guizhou in high incidence season, grippotyphosa and bataviae are the newly discovered serogroups of Leptospira in Guizhou.

Objective:To explore the effect of early quantitative pulmonary rehabilitation assessment in adult Intensive Care Unit (ICU) patients with mechanical ventilation in high altitude area.Methods:From March 2019 to October 2021, convenience sampling was used to select 287 adult ICU patients with mechanical ventilation of Qinghai Red Cross Hospital as the research object. According to the time of admission, the patients were divided into the control group (142 cases) and the experimental group (145 cases) . The control group was given the routine pulmonary rehabilitation, and the experimental group received the early pulmonary rehabilitation based on quantitative assessment. The Acute Physiology and Chronic Health EvaluationⅡ (APACHEⅡ) score and the Intensive Care Units Mobility Scale (IMS) score were compared between the two groups before enrollment, on the eighth and sixteenth days of pulmonary rehabilitation. The oxygenation index of the two groups of patients before enrollment and on the first, fourth, sixth, eighth and sixteenth days of pulmonary rehabilitation, the time of ICU stay, the time of mechanical ventilation, the success rate of ventilator removal and the complications of the two groups of patients with mechanical ventilation were also compared.Results:On the eighth and sixteenth days of pulmonary rehabilitation, the APACHE Ⅱ score of the experimental group was lower than that of the control group, and the IMS score was higher than that of the control group, with statistical differences ( P<0.05) . On the sixth, eighth and sixteenth days of pulmonary rehabilitation, the oxygenation index of the experimental group was higher than that of the control group, and the difference was statistically significant ( P<0.05) . The ICU stay time and mechanical ventilation time in the experimental group were lower than those in the control group, and the success rate of ventilator removal in the experimental group was higher than that in the control group, with statistical differences ( P<0.05) . Conclusions:Implementing early pulmonary rehabilitation for adult ICU patients with mechanical ventilation in high altitude area is conducive to promoting pulmonary rehabilitation of patients, improving the success rate of ventilator removal, and reducing patients' ICU stay time, mechanical ventilation time and the occurrence of complications.

Objective:To explore the effect of early quantitative pulmonary rehabilitation assessment in adult Intensive Care Unit (ICU) patients with mechanical ventilation in high altitude area.Methods:From March 2019 to October 2021, convenience sampling was used to select 287 adult ICU patients with mechanical ventilation of Qinghai Red Cross Hospital as the research object. According to the time of admission, the patients were divided into the control group (142 cases) and the experimental group (145 cases) . The control group was given the routine pulmonary rehabilitation, and the experimental group received the early pulmonary rehabilitation based on quantitative assessment. The Acute Physiology and Chronic Health EvaluationⅡ (APACHEⅡ) score and the Intensive Care Units Mobility Scale (IMS) score were compared between the two groups before enrollment, on the eighth and sixteenth days of pulmonary rehabilitation. The oxygenation index of the two groups of patients before enrollment and on the first, fourth, sixth, eighth and sixteenth days of pulmonary rehabilitation, the time of ICU stay, the time of mechanical ventilation, the success rate of ventilator removal and the complications of the two groups of patients with mechanical ventilation were also compared.Results:On the eighth and sixteenth days of pulmonary rehabilitation, the APACHE Ⅱ score of the experimental group was lower than that of the control group, and the IMS score was higher than that of the control group, with statistical differences ( P<0.05) . On the sixth, eighth and sixteenth days of pulmonary rehabilitation, the oxygenation index of the experimental group was higher than that of the control group, and the difference was statistically significant ( P<0.05) . The ICU stay time and mechanical ventilation time in the experimental group were lower than those in the control group, and the success rate of ventilator removal in the experimental group was higher than that in the control group, with statistical differences ( P<0.05) . Conclusions:Implementing early pulmonary rehabilitation for adult ICU patients with mechanical ventilation in high altitude area is conducive to promoting pulmonary rehabilitation of patients, improving the success rate of ventilator removal, and reducing patients' ICU stay time, mechanical ventilation time and the occurrence of complications.

Objective To understand the genetic and epidemiologic characteristic of Brucella (B.) melitensis strains isolated in Guizhou province in 2010-2012. Methods B. genus specific BCSP31-PCR and species-specific AMOS-PCR were used to identify the bacteria strain,while the identified strains were analyzed under MLVA-16 and cluster analysis of B. melitensis strains. The strains were isolated from Guizhou and other provinces. Results Six B. melitensis strains were identified as B. melitensis using the BCSP31-PCR and AMOS-PCR. Data from the MLVA-16 analysis revealed the differences of repeated numbers at parts of the VNTR locus in the six strains isolated in Guizhou province. The six strains from Guizhou province and 105 B. melitensis strains from other province could be divided into 72 MLVA types(MT). Strain ZY and ZA from Guizhou province were typed as MT63,and LL3,LL4 and LL11 were typed as MT67,while strain SQ was typed as MT72. Data from the clustering analysis showed that ZY,ZA,LL3,LL4 and LL9 were most closely clustered with B. melitensis isolates from Yunnan,Fujian and Guangdong provinces,but strain SQ was genetically remote from other strains. Conclusion PCR methods,combined with MLVA-16, identified the six B. melitensis strains isolated in Guizhou province in 2010-2012 as B. melitensis biovar 3,with the genetic diversity of the strains showed. Six strains were closely related to the B. melitensis strains from Yunnan,Fujian and Guangdong provinces. The results of this study provided scientific basis for the control and prevention of Brucellosis in Guizhou province.