Objective To investigate the distribution and sequence conservation of genes encoding the outer membrane lipoprotein D(LPD)of nontypeable Haemophilus influenzae(NTHi)isolates and to ana-lyze the immunogenicity and the immunoprotective effects of the expressed recombinant LPD(rLPD). Meth-ods PCR analysis was used to detect the genes encoding LPD of NTHi isolates. The PCR products were se-quenced after T-A cloning. A prokaryotic expression system for genes encoding LPD was established to ex-press the rLPD. Ni-NTA affinity chromatography was used for purification. SDS-PAGE and Bio-Rad Gel Im-age Analyzer were used to detect the expression and the yield of rLPD. The antigenicity and immunoreactivity of rLPD were detected by ELISA and Western blot assay. The immunoprotective effects of rLPD against lethal dose of NTHi were evaluated in a mouse model. Results All of the tested NTHi isolates were positive for the genes encoding LPD. They shared 98. 0% -99. 4% homologies in nucleotide sequences and 98. 5% -100% homologies in amino acid sequences. The established prokaryotic expression system expressed rLPD with a high yield. High levels of antibody in rabbits were induced by the rLPD. The anti-NTHi antiserum samples from rabbits and children could recognize and react with the rLPD. The result of ELISA indicated that 93. 6%(58 / 62)and 53. 2%(32 / 62)of the serum samples from children with NTHi infection were positive for rLPD-IgM and rLPD-IgG,respectively. The rLPD at concentrations of 100 μg and 200 μg could respectively protect 60. 0% and 73. 3% of mice from lethal NTHi infection. Conclusion The genes enco-ding LPD were extensively distributed in NTHi isolates with high sequence conservation. The expressed rLPD could be used as a potential candidate antigen in the development of genetic engineering vaccine against NTHi infection considering its high immunogenicity and immunoprotective effects.

Objective To observe the clinical effect of the maintenance for the liver and kidney function by extra corporeal membrane oxygenation (ECMO) in brain death donor with severe hemodynamic instability.Methods Ninety-nine brain death donors maintained by ECMO were followed up.The criteria for using the ECMO to protect the organ function were as follow:cardiopulmonary resuscitation history (cardiac compression ＞ 20 min);mean arterial pressure (MAP),for Adult ＜60-70 mmHg,for child ＜50-60 mmHg,and for infant ＜40-50 mmHg;cardiac index ＜2 L/(m2 ·min) (3 h);Large doses of vasoactive drugs,for doparnine 20μg/(kg·min),for (norepinephrine) epinephrine 1.0 μg/(kg· min) (3 h),and for oliguria ＜0.5 mL/(kg · h);blood biochemical indexes,moderate,severe impairment on acute hepatic and renal function;others,ST-T significant changes in electrocardiogram,and difficult to correct the metabolic acidosis (3 h).The organs were evaluated during their retrieval and as well their evolution after transplantation was evaluated.Results ECMO allowed for the maintenance of hemodynamic stability before organ procurement.A total of 99 cases receiving ECMO maintenance were collected,equal to100 ％ of the total donation cases (100％).198 kidneys,and 99 livers were procured from these donors meanwhile 15 kidneys and 42 livers respectively were discarded as theywere shown in a macroscopic evaluation.177 of the procured kidneys were transplanted.DGF of kidney transplantation was observed in 20.9％of the cases.Acute rejection incidence was 12.99％.Transplanted kidneys and recipient survival rate was 96.1％/99.3％ for one year,94.7％/97.8％ for 3 years,and 93.6/97.8％ for 4 years,respectively.There was no significant difference in patient or graft survival between the group with ECMO and the group without ECMO.Conclusion ECMO in the brain dead donors with severe circulatory dysfunction allows to avoid organ donors loss and obtain good quality kidneys and livers with excellent graft survival after transplantation.