Objective To study the effects of D-Ala2-Leu5-enkephalin(DADLE) preconditioning on rabbit donor heart preservation and protective mechanism.Methods 24 rabbits were randomly divided into 4 groups. In group B, after preconditioning with DADLE(1mg?kg -1 ), the donor hearts were then arrested and preserved with Krebs-Henseleit buffer at 4℃ for 4h,and group A without conditioning as controls. Then the donor hearts were transplanted into the abdomen of recipient rabbits. Recovery situation of contraction of the donor heart was compared. The left ventricular tissues were obtained from the donor hearts after 2h of transplantation.The contents of free radicals and ATPase activity were determined and cardiac ultrastructure were observed.Results After 4h cold storage, the heart undergoing DADLE preconditioning in group B showed better recovery of left ventricular development pressure (LVDP).The activity of sodium-potassium ATPase in group B were higher than that in group A. DADLE increased free radical production 2-fold versus group A. Group B showed slighter injury in transmission electron microscopy observation than that in group A.Conclusion Our study demonstrated opioid preconditioning has protective function to rabbit heart injury caused by long term cold storage. The protective mechanism maybe related to increase of free radical content.

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Objective To evaluate the effects of flurbiprofen axetil administered at different time points on oxygenation in the patients undergoing one-lung ventilation (OLV).Methods Ninety patients of both sexes,aged 45-64 yr,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,undergoing elective thoracoscope-assisted pulmonary lobectomy,were assigned into 3 groups (n =30 each) using a random number table:control group (group C),preoperative administration group (group F1) and intraoperative administration group (group F2).Flurbiprofen axetil (10 mg/ml) and fat emulsion 10 ml were injected intravenously at 15 min before operation in F1 and C groups,respectively.Flurbiprofen axetil 10 ml was intravenously injected immediately after the beginning of OLV in group F2.At 15 min before operation (T1),15 and 30 min of OLV (T2,3),and 15 min after restoration of two-lung ventilation (T4),airway peak pressure (Ppeak) and dynamic lung compliance (Cdyn) were recorded,arterial blood samples were collected for blood gas analysis.The arterial oxygen partial pressure (PaO2) was recorded,and the oxygenation index (OI) and intrapulmonary shunt (Qs/Qt) were calculated.The concentrations of thromboxane B2 (TXB2) and 6-keto-prostaglandin F1α (6-K-PGF1α) in serum were measured by enzyme-linked immunosorbent assay,and TXB2/6-K-PGF1α ratio was calculated.The development of interrupting OLV due to SpO2＜90％ and postoperative dyspnea,pulmonary infection,atelectasis and length of hospital stay were recorded.Results Compared with group C,PaO2 and OI were significantly increased,and Qs/Qt was decreased at T2,3,the serum concentrations of TXB2 and 6-K-PGF1α were decreased,and TXB2/6-K-PGF1α ratio was increased at T2-4,the incidence of interrupting OLV was decreased (P＜0.05),and no significant change was found in the parameters mentioned above in group F2 (P＞0.05).Compared with group F1,PaO2 and OI were significantly decreased at T2,3,Qs/Qt was increased at T2,and the serum concentrations of TXB2 and 6-K-PGF1α were increased,and TXB2/6-K-PGF1α ratio was decreased at T2-4 in group F2 (P＜0.05).There was no significant difference in the incidence of postoperative dyspnea,pulmonary infection and atelectasis and length of hospital stay between the three groups (P＞0.05).Conclusion Flurbiprofen axetil injected at 15 min before operation can significantly improve oxygenation and prevent the development of hyoxemia in the patients undergoing OLV,however,flurbiprofen axetil administered immediately after the beginning of OLV has no such effect.

The use of the breathing machine undoubtedly represents the progress in the field of respiratory insufficien-cy treatment,although it has saved many lives,but its side effects are also as everyone knows. Ventilator-associated pneumonia is the most important complication of mechanical ventilation ,it can make the cost of hospitalization and the mortality of patients increased significantly. Pathological biopsy and histological training as the “gold standard ”for di-agnosis of ventilator-associated pneumonia ,but it is the invasive operation and it difficult to fully implement , recently the diagnosis is still controversial. Emerged in recent years the biomarkers for the diagnosis research , but the diver-gence of the corresponding research conclusion is wide. Now , biomarkers that would be useful for diagnosis of ventila-tor-associated pneumonia are reviewed so as to it can improve people ’s understanding of the value of biomarkers in the diagnosis of ventilator-associated pneumonia.

Objective To construct lentiviral vector of late endosomal／lysosomal adaptor, mitogen-activated protein kinase and mammalian target of rapamycin activator 2 ( lamtor2) gene, and to explore its regulatory role on inflammatory response of macrophages after Klebsiella pneumoniae infection.Methods Two pairs of mouse lamtor2 short hairpin RNA (shRNA) were designed and sub-cloned into PLKO.1-puro to construct lentiviral vector, and were transfected into the murine RAW264.7 macrophage.There were two experimental groups including pLKO.1-puro shlamtor 2-1(sh1 group) and pLKO.1-puro shlamtor 2-2 (sh2 group), and the RAW264.7 cells transfected with non-treated pLKO.1-puro was set as control.The expession of lamtor2 were detected by real-time quantitative polymerase chain reaction ( RT-qPCR ) and Western blot.The levels of inflammatory factors including interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-αsecreted by the cells were detected by RT-qPCR.T test was used for comparison between groups.Results The recombinant lentiviral vector PLKO.1-shlamtor 2 transfected RAW264.7 cells successfully.The relative expressions of lamtor2 mRNA in the control group, the sh1 group and the sh2 group were 1.000 ±0.000, 0.596 ±0.125 and 0.120 ±0.080, respectively.The expression of lamtor2 in the sh2 group was significantly lower than that in the sh 1 group (t＝3.399, P＝0.015), and they were both significantly lower than the control group ( t ＝3.333 and 9.734, respectively, both P ＜0.05).After infection with Klebsiella pneumoniae, expression levels of IL-1β( t ＝15.20), IL-6 (t＝43.30) and TNF-α(t＝12.67) were significantly higher than those in the control group (all P＜0.01).Conclusion The lentiviral vector of lamtor2 can stably down-regulate the expression of lamtor2 gene in macrophages through RNA interference mechanism , which has a significant effect on the secretion of inflammatory factors of macrophages that are infected with Klebsiella pneumoniae.

Objective: To construct lentiviral vector of late endosomal/lysosomal adaptor, mitogen-activated protein kinase and mammalian target of rapamycin activator 2 (lamtor2) gene, and to explore its regulatory role on inflammatory response of macrophages after Klebsiella pneumoniae infection. Methods: Two pairs of mouse lamtor2 short hairpin RNA (shRNA) were designed and sub-cloned into PLKO.1-puro to construct lentiviral vector, and were transfected into the murine RAW264.7 macrophage. There were two experimental groups including pLKO.1-puro shlamtor 2-1(sh1 group) and pLKO.1-puro shlamtor 2-2 (sh2 group), and the RAW264.7 cells transfected with non-treated pLKO.1-puro was set as control. The expession of lamtor2 were detected by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot. The levels of inflammatory factors including interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α secreted by the cells were detected by RT-qPCR. T test was used for comparison between groups. Results: The recombinant lentiviral vector PLKO.1-shlamtor 2 transfected RAW264.7 cells successfully. The relative expressions of lamtor2 mRNA in the control group, the sh1 group and the sh2 group were 1.000±0.000, 0.596±0.125 and 0.120±0.080, respectively. The expression of lamtor2 in the sh2 group was significantly lower than that in the sh1 group (t=3.399, P=0.015), and they were both significantly lower than the control group (t=3.333 and 9.734, respectively, both P< 0.05). After infection with Klebsiella pneumoniae, expression levels of IL-1β (t=15.20), IL-6 (t=43.30) and TNF-α (t=12.67) were significantly higher than those in the control group (all P<0.01). Conclusion The lentiviral vector of lamtor2 can stably down-regulate the expression of lamtor2 gene in macrophages through RNA interference mechanism, which has a significant effect on the secretion of inflammatory factors of macrophages that are infected with Klebsiella pneumoniae.