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Objective:To evaluate the clinical value of intraoperative transesophageal echocardiography (TEE) in monitoring left ventricular assist device (LVAD) implantation.Methods:Between March 2019 and November 2021, 23 consecutive patients from TEDA International Cardiovascular Hospital, including 21 cases with dilated cardiomyopathy, 1 case with myocardial noncompaction and 1 case with ischemic cardiomyopathy, underwent HeartCon blood pump, a type of third generation LVAD implantation for severe heart failure. TEE was preformed in all cases before and after cardiopulmonary bypass. The dimensions of left-sided and right-sided cardiac chamber, ventricular function, de-airing, interventricular septal position, inlet cannulae position and the function of device were observed and recorded during LVAD implantation. Paired t test was used for statistical analysis of left-sided and right-sided heart parameters in pre- and post-operative measurements. Results:The left heart was dilated significantly and coexistent with right heart enlargement in some degree before LVAD implantation in total 23 cases. More than moderate mitral regurgitation (MR) in 16 cases and less than moderate MR in 7 cases were present. Mild or trace aortic regurgitation (AR) existed in 13 cases. More than moderate tricuspid regurgitation (TR) in 4 cases and less than moderate TR in 16 cases were observed. Left atrial appendage thrombosis was detected in 2 cases. After LVAD implantation, TEE revealed that the left ventricular end-diastolic diameter reduced significantly (42 mm/m 2 vs 32.8 mm/m 2, P<0.05) and left ventricular ejection fraction increased accordingly (22.2% vs 34.0%, P<0.05). There were no significant differences in right ventricular diameter and fractional area change between pre- and post-operative findings(all P>0.05). The ratio of left ventricular inner diameter to right ventricular inner diameter (2.09 vs 1.69, P<0.05) decreased in total 23 cases after LVAD implantation.Interventricular septal position became neutral position instead of pre-oprative rightward position. The severity of MR decreased in varying degrees in total 23 cases after LVAD implantation. All patients underwent tricuspid valvuloplasty with residual mild regurgitation in 8 cases. Conclusions:HeartCon blood pump can effectively unload the left ventricle with sufficient cardiac output in patients with severe congestive heart failure. TEE plays a major role in the clinical decision making during LVAD implantation, which can evaluate pre-operative cardiac abnormalities, intra-operative air embolism, inlet cannulae position, cannulas patency and cardiac function, especially blood volume status and the balance between double ventricles, which is critical for optimal functioning of the device.
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Objective:To investigate and evaluate the efficacy of HeartCon left ventricular assist device (LVAD) in the treatment of adult patients with end-stage heart failure (ESHF).Methods:A prospective and observational study was conducted. Patients with ESHF who underwent LVAD implantation in the department of cardiac surgery of Teda International Cardiovascular Hospital from September 2020 to August 2021 were selected. The left ventricular end-diastolic diameter (LVEDD), left ventricular ejection fraction (LVEF), New York Heart Association (NYHA) classification, N-terminal pro-B type natriuretic peptide (NT-proBNP), and six minute walk distance (6MWD) before operation and 90 days after operation were compared. The incidence of equipment failures and major adverse events within 90 days after operation were recorded.Results:A total of 20 patients with ESHF were included, with 15 males and 5 females. Patients' age ranged from 20 to 67 years old, with an average of (50.2±13.6) years old. The range of body weight was 49.8-106.1 kg, with an average of (67.9±15.5) kg, and the body surface area (BSA) was from 1.49 to 2.32 m 2, with an average of (17.6±0.22) m 2. The operation process of all the patients were successful. The length of hospital stay ranged from 33 to 90 days, and the average was 56.0 (42.8, 75.0) days. Complications within 90 days after operation as follows, 2 cases with pericardial tamponade (10%), 1 case with cerebral hemorrhage (5%), 1 case with mediastinum infection (5%), 3 cases with acute renal injury (AKI, 15%), 5 cases with gastrointestinal bleeding (25%). There were no mechanical failure of LVAD and hemolysis events, right ventricular failure (RVF), cerebral infarction and death occurred. Compared with preoperative, the LVEDD significantly decreased (mm: 67.50±13.98 vs. 77.40±9.73), LVEF significantly increased (%: 34.80±9.76 vs. 22.70±5.62), NT-proBNP significantly decreased (ng/L: 2 028.65±1 752.05 vs. 4 796.45±4 355.40), 6MWD significantly increased (m: 385.20±144.12 vs. 85.81±63.50) at 90 days after operation, and the differences were statistically significant (all P < 0.05). 18 cases (90%) of the 20 patients reached NYHA classification Ⅰ and 2 cases (10%) reached NYHA classification Ⅱ, which were significantly improved compared with those before surgery (all patients' NYHA classification were Ⅳ before surgery). Conclusion:HeartCon LVAD can effectively improve the life quality of patients with ESHF, which has been proved safe and effective in clinical trials, but its long-term effects and complications need further observation and study.
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Objective:To observe the risk factors of acute renal injury(AKI) after coronary artery bypass grafting(CABG) and the influence of blood pressure on AKI.Methods:980 patients in CABG of Cardiology Department of TEDA International Cardiovascular Hospital were diagnosed with AKI according to the AKIN standard, with 706 males and 274 females, averaged(61.9±8.0)years old. Patients were divided into two groups according to whether AKI occurred: AKI group(86 cases) and non AKI group(894 cases). The baseline clinical data, operation related data were compared between the two groups. At the same time, according to the preoperative mean systolic blood pressure(SBP) level, LSP[mean systolic blood pressure<120 mmHg(1 mmHg=0.133 kPa), 374 cases], MSP(mean systolic blood pressure 120-140 mmHg, 481 cases) and HSP(mean systolic blood pressure≥140 mmHg, 125 cases) were classified as covariates, and the influencing factors of dependent variable AKI were analyzed by multivariate logistic regression.Results:The prevalence of AKI was 8.7%(86/980). Compared with non-AKI group, preoperative SBP[(129.8±13.8)mmHg vs.(124.4±13.3)mmHg, P=0.000], mean arterial pressure[(91.9±8.8)mmHg vs.(88.8±9.1)mmHg, P=0.004], and mean pulse pressure[(56.9±10.7)mmHg vs.(53.2±9.8)mmHg, P=0.001]were increased significantly. After adjusted for other risk factors, preoperative SBP elevation, hypertension history, cardiopulmonary bypass(CPB), use of intra-aortic-balloon-pump(IABP), secondary thoracotomy, preoperative diuresis, intraoperative blood transfusion and baseline low glomerular filtration rate(eGFR) were independent risk factors for AKI after CABG. Compared with LSP group, the relative risk of AKI after CABG in HSP group was 2.743(95% CI: 1.595-4.715). In patients with hypertension history, AKI in HSP group was significantly higher than that in LSP group(18.4% vs. 8.1%, P=0.001). However, the preoperative blood pressure level of patients who denied the history of hypertension had no effect on AKI. Conclusion:Preoperative SBP is a risk factor for AKI after CABG. The incidence of AKI after CABG can be significantly reduced by controlling SBP below 140 mmHg in patients with hypertension.
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Objective To evaluate the effect of Levosimendan on the prognosis in patients with severe coronary heart disease after operation.Methods A total of 485 severe coronary disease patients undergoing coronary artery bypass grafting from Teda International Cardiovascular Hospital and the Cardiac Surgery Department of the First Affiliated Hospital of China Medical University from May 2014 to June 2016 were enrolled.Of them,45 cases receiving Levosimendan postoperatively were assigned to the Levosimendan group,and according to propensity score matching,another 45 cases were selected as the control group in this study.Clinical data before treatment had no difference between the groups (P > 0.1).Postoperative prognosis was compared between the two groups.Results There were significant differences in heart rate,mean arterial pressure,central venous pressure,cardiac output and other hemodynamic parameters between the two groups 48h after operation.The heart ultrasound results showed that the left ventricular ejection fraction(IVEF) was increased [(0.53±0.12) %vs.(0.46±0.09)%,t =2.594,P=0.002],the postoperative ventilation time was reduced [(46.8±11.3) h vs.(58.5±16.3) h,t=-2.031,P=0.045]and the onset of bowel sounds became early [(16.5±5.9) h vs.(18.7±10.1) h,t =1.592,P=0.039]in the levosimendan group than in the control group 48h after operation.The incidences of new-onset acute kidney injury(20 % and 40 %,x2 =6.702,P =0.018),new-onset postoperative atrial fibrillation (15.6% and 44.4%,x2 =6.156,P =0.023) and perioperative myocardial infarction(11.1 % and 33.3%,x2 =6.429,P =0.021) had significant differences between the two groups(P<0.05),but there was no difference in ICU retention time,1-month mortality after operation,malignant arrhythmia incidence and auxiliary equipment use (P > 0.05).Conclusions Levosimendan can improve the early prognosis of severe coronary disease patients undergoing coronary artery bypass grafting and reduce the occurrence of postoperative organ dysfunction.
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Objective To analyze the risk factors and pathogen distribution of ventilator-associated pneumonia (VAP) in patients following coronary artery bypass grafting (CABG).Methods The clinical data of 1414 patients undergoing CABG in Teda International Cardiovascular Hospital from June 2014 to June 2016 were retrospectively analyzed .The VAP developed following CABG in 42 patients ( VAP group ) and not occurred in 1372 patients ( non-VAP group) .Multivariate logistic regression was used to analyze the risk factors of VAP in patients after CABG , and the microbial culture and drug sensitivity test were performed in VAP patients.Results The incidence rate of VAP after CABG was 2.97% (42/1414).Multivariate logistic regression analysis showed that history of smoking ( OR =2.216, 95% CI 1.018 -4.825, P <0.05), mechanical ventilation time >48 h (OR=7.457, 95% CI 3.443-16.161, P<0.01), LVEF<40%(OR=3.524 , 95% CI 1.203-10.325, P<0.05) and postoperative acute kidney injury (OR=16.239, 95% CI 7.551 -34.924, P <0.01) were independent risk factors for VAP in patients after CABG.A total of 42 pathogen strains were detected in 42 patients with VAP, including 37 strains of Gram-negative bacteria, 2 strains of Gram-positive bacteria, and 3 strains of fungus.Gram-negative bacteria mainly were Klebsiella pneumoniae subspecies ( n =23, 54.76%) and Burkholderia cepacia ( n =6, 14.27%);the Gram-positive bacteria were Staphylococcus aureus ( n =2, 4.76%);the fungus was Candida albicans ( n =3, 7.14%).Klebsiella pneumoniae was sensitive to many antibiotics;and the resistance rate to amikacin , aztreonam , meropenem , and levofloxacin was <10%, the resistance rate to ceftazidime and piperacillin was <25%. Burkholderia cepacia was naturally resistant to amikacin , ampicillin, aztreonam, cefazolin, gentamicin and sulfamethoxazole .Conclusion The incidence of VAP was higher in patients after CABG , and the involved pathogens were mainly Gram-negative bacteria .Clinically , it is necessary to take necessary measures to prevent and treat VAP in order to improve the prognosis of patients undergoing CABG .
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Objective To investigate the antagonism of third generation dihydropyridine calcium channel antagonist beni-dipine in contraction induced by potassium chloride(KCl)in human internal mammary artery(IMA),and in order to explore the possibility of using benidipine as an antispastic agent during coronary artery bypass grafting(CABG). Methods The IMAs were taken from 19 patients undergoing CABG. The vasodilator effect(n = 7)on KCl-induced contraction and the inhibition effect(n = 6)after incubation with benidipine were studied in a myograph. The caveolin 1. 2 was detected(n = 3,2 cases re-spectively)by enzyme-linked immunosorbent assay (ELISA). Results The maximum relaxation caused benidipine was (87. 7 ± 4. 9)% . Incubation with benidipine significantly depressed the contraction by KCl[from( 16. 9 ± 4. 4)mN to (9. 1 ± 3. 4 )mN,P < 0. 01]. The concentration of caveolin 1. 2 was down-regulated after incubation with benidipine(P < 0. 05). Conclusion Benidipine has a potent inhibitory effect on KCl-induced vasoconstriction. The relaxation may be related with the expression of caveolin 1. 2. Using benidipine in patients undergoing CABG may provide antispastic effects in grafts.
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Innovation education was introduced in medical microbiology teaching practice,including updating the innovative education concept,reforming teaching methods and means,constructing the teaching content system and practice platform adapting to innovation education.
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Objective To discuss clinic effect of pulmonary artery valvuloplasty for right ventricle outflow tract (RVOT) reconstruction in patients with repaired tetralogy of fallot (TOF). Methods A total of 93 cases TOF were randomly divided into Group A (n = 49) or Group B (n = 44) to perform the procedure with repaired tetralogy of fallot Group A received autogenous pericardium to enlarge RVOT and pulmonary valvuloplasty.Group B received autogenous pericardium to enlarge RVOT by routine therapy. Pulmonary regurgitation index (PRi) and the ratio between pulmonary regurgitant jet width and pulmonary annulus diameter were measured with echoeardiography. Results The mean follow-up was 3. 1 ± 0. 2 years. The PRi and the ratio between pulmonary regurgitant jet width and pulmonary annulus diameter in Group A were-significantly lower than Group B (0. 55 ± 0. 13 vs. 0. 61 ± 0. 10, t = 2. 685, P < 0.01) and ([52.0 ± 10.4] % vs. [57.1 ± 10. 5]% ,t = 2. 349, P < 0.05) . Three-dimensional ultrasound examination showed that 69% (34/69) of pulmonary valves in Group A was developed well. Conclusion Pulmonary valvuloplasty during transannular patch for repaired TOF may prevent fre'e pulmonary regurgitation and can obtain good clinical outcome.
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AIM: To explore the effect of complement on the cerebral ischemia/reperfusion injury in rat and the protection by sCR1-SCR15-18. METHODS: 75 male SD rats were randomly divided into three groups: sham operation group (SO, n=15), middle cerebral artery occlusion and reperfusion (MCAO) without treatment group (I/R, n=30); MCAO treated with sCR1-SCR15-18 group (sCR1-SCR15-18, n=30). After the MCAO for 2 h, then reperfusion for 24 h, the scores of neural behavioral functional deficits were determined. Infarction area was measured by TTC staining. Activity of MPO in cerebral cortex was detected. C3b deposition and pathological change were observed by immunohistochemial staining and HE staining, respectively. RESULTS: After reperfusion for 24 h, the neurological deficits score, infarction area and activity of MPO in sCR1-SCR15-18 group were decreased compared to I/R group. In sCR1-SCR15-18 group, C3b deposition in ischemic area was decreased and pathological injury was improved compared to I/R group. CONCLUSION: Complement plays a role in cerebral ischemia-reperfusion injury and sCR1-SCR15-18 exerts a protective effect by inhibiting the excessive activation of complement.
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Objective To construct and express a small complement receptor type 1 (CR1) deriv-ative which contained two functional domains. Methods Total RNA was isolated from the peripheral blood mononuclear cell. The functional fragment Ⅰ of CR1 was amplified using the RT-PCR. The functional frag-ment Ⅱ was amplified with the plasmid of pET-32a-CR1-SCR15-18 as template which had been already con-structed in our laboratory. Then the chimeric gene that contained the two fragments was constructed with spli-cing overlap extention PCR. The chimeric gene was then inserted into the plasmid of pET-32a (+) and transformed into E. coli Rosetta(DE3). The inserted gene was verified by enzyme digestion and DNA se-quencing. After induced by IPTG, the expressed protein was analyzed by SDS-PAGE and Western blot. Then the fusion protein was purified by Ni-NTA affinity column and renatured through dialysis. The comple-ment inhibition activity was determined by CH50 method. Results The chimeric gene was successfully cloned into pET-32a(+). The result of SDS-PAGE and Western blot confirmed the expressed protein and showed that the molecule mass(Mr) of the expressed protein was 63×103. The purity of the recombinant protein was up to 92% after Ni-NTA column affinity chromatography. The bioactivity assay showed the fusion protein had a concentration-dependent complement inhibition activity within the concentration range of 0-200 μg/ml. Conclusion The two functional domains contained small CR1 derivative was successfully construc-ted and expressed in E. coli Rosetta. The fusion protein had a relative high bioactivity, providing a basis for further function experiment in vivo.
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Objective: To screen the best genetic engineering bacterium for the production of peptide antibiotic hPAB-? and evaluate its fermentation level in bottle. Methods:After analysis of the interest fusion protein expression levels of 8 recombinant bacteria containing 1-8 copies of human peptide antibiotic hPAB-? expressing plasmid respectively,2-5 copies expressing bacteria were chosen for the further study of their bacteria yield,expression forms of the target protein, dissolution of the inclusion bodies and the efficiency of fusion protein purification by affinity chromatography, then the best engineering bacterium with the certain copies of interest peptide expressing plasmid was screened out and its optimal fermentation parameters in bottle were also studied. Results:The recombinant bacterium transformed by 3 copies of interest peptide expressing plasmid was the best candidate for its bacteria yield (3.153 g/L) and fusion protein expression level (27.7%) were the highest among 1-8 copies candidates. The inclusion bodies of 3 copies target fusion protein could be easily dissolved by 8 mol/L urea and captured by Ni-NTA column. The elution of the fusion protein could be directly cleaved to monomer by adding 2 mol/L hydroxylamine, adjusting pH to 9.0 and incubating at 45℃ for 2 h. The optimal fermentation conditions of the selected recombinant bacteria were: culture the organisms with modified M9-CAA media at 37℃ and 160 r/min to (A 600 )≈2.5, then add IPTG to the final concentration 100 ?mol/L to induce the expression of target fusion protein for 5 h. Conclusion:The engineering bacterium containing 3 copies interest peptide recombinant expressing plasmid is the best candidate for the production of peptide antibiotic hPAB-?,and its fermentation parameters are confirmed.
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Objective To study the roles of protein kinase C (PKC) in the signal transduction pathway of LPS induced nuclear transcription factor (NF) ?B activation of alveolar macrophages(AMs). Methods NF ?B level in nuclear protein extraction of AMs was detected by sandwich ELISA. Intranuclear translocation of NF ?B was observed by immunocytochemical staining. Results LPS could induce NF ?B activation of AMs in time and dose dependent manners. Immunohistochemical staining revealed intranuclear translocation following LPS induction. Specific calphostin C (Cal C) and pyrrolidine dithiocarbamate (PDTC) could inhibit LPS induced NF ?B activation. Conclusion PKC, as a up stream messenger, is involved in the signal transduction pathway of LPS induced NF ?B activation.
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Objective To clone and express the gene encoding the binding domain of human soluble complement receptor type 1 (sCR1). Methods sCR1-SCR15-18 cDNA was amplified using RT-PCR from human monocytes of peripheral blood and sequenced using vector pMD-18T. Recombinant pET32-sCR1-SCR15-18 was constructed using prokaryotic expression vector pET32 and transformed into bacterium BL21. IPTG was used to induce gene expression and the obtained expression product was identified by immunoblotting. Results The gene segment that specifically encodes sCR1 was synthesized, the sequence of which was consistent with that of sCR1-SCR15-18 cDNA as registered at GenBank. A prokaryotic expression recombinant pET32-sCR1-SCR15-18 was constructed. The amount of target protein accounted for 40% of the total bacterial proteins and inclusion bodies were present in the bacteria. Immunoblotting showed a single positive band at the site of 43?10~(3). Conclusion The gene encoding sCR1-SCR15-18 was cloned from human monocytes and efficiently expressed in E.coli.
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Objective To prepare highly expressed, purified and refolded SCR15-18 of human soluble complement receptor type 1 (sCR1-SCR15-18) protein. Methods The expression of recombinant pET32-sCR1-SCR15-18 in E.coli.. BL21 was induced by IPTG of different concentrations for different time period under different temperatures and the bacteria were split by sonication. The sCR1-SCR15-18 protein was purified by Ni2+-NTA resin affinity chromatography. The purified protein was refolded under different conditions. Then the bioactivity of the protein was analyzed. Results The sCR1-SCR15-18 protein of high expression, purity and bioactivity was attained. Conclusion The parameters of expression, purification and refolding of sCR1-SCR15-18 protein were optimized, which may pave a way for further studies.
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AIM: To investigate the influences of bypass-activated complement and TNF-? on intercellular adhesion molecule 1(ICAM-1) expression in endothelial cells, and polymorphonuclear neutrophil-endothelial cell (PMN-EC) adhesion. METHODS: In vitro, zymosan-activated human serum(ZAHS) and/or TNF-? directly stimulated the HUVECS monolayers. PMN-EC adhesion percentage was measured, and modified whole-cell ELISA was used for measurement of ICAM-1. RESULTS: ZAHS alone resulted in no significant change in the degree of PMN adhesion and the level of ICAM-1. Activation of HUVECS with TNF-? followed by ZAHS stimulation resulted in greater increase in PMN-EC adhesion and ICAM-1 expression, as compared with the activation with TNF-? alone. CONCLUSION: Bypass-activated complement can promote ICAM-1 expression and neutrophil -endothelium adhesion induced by TNF-?, and so it is effective for promoting inflammation.
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Objective To express the short consensus repeat 15-18 (SCR15-18) domain of human complement receptor 1 (CR1) in Pichia pastoris as a secreted protein in order to found a base for large-scale industry fermentation. Methods The gene fragment of CR1-SCR15-18 was amplified by PCR from plasmid pET32a-sCR1-SCR15-18. The obtained sequence was subcloned into Pichia pastoris secretory expression vector pPIC9. After identification the recombinant plasmid pPIC9-CR1-SCR15-18 was electrotransported into the yeast. Positive clones were identified using colony PCR. Positive recombinants were fermented in shake flaskes and induced with methanol. The recombinant proteins were identified with SDS-PAGE and Western blot analysis. After the recombinant protein was purified by Ni-NTA agarose metal chelate affinity chromatography, inhibiting complement hemolysis testing was used to detect the biological activity. Results Recombinant expression plasmid pPIC9-CR1-SCR15-18 was successfully constructed. SDS-PAGE and Western blot analysis revealed that the target gene was successfully expressed in the yeast and the recombinant protein was successful secreted in the culture supernatant. After purification, the protein inhibited complement hemolysis in vitro. Conclusion CR1-SCR15-18 is successful expressed in Pichia Pastoris with high activity of inhibiting complement hemolysis.
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In order to verify that bypass-activated complement can enable polymorphonuclear neutrophils (PMN) to lose their bactericidal capacity,3 experiments were performed as follows:(DA monolayer culture of human PMN was added with zymosan-activated human serum (ZAHS).then the superoxide ion (O2-,the specific granules,and the intracellular bactericidal activity of the cultured PMNs were signigicantly decreased and reached the lowest point in the 6th hour after the adding of ZAHS.0.05 ml of ZAHS showed the maximal effects on the PMNs.(2)Mice were injected intravenously with 0.5 ml of ZAHS and were killed 6 hours later.All the 3 parameters mentioned above dropped markedly in the PMNs isolated from the blood and the lungs of the mice.Pathologically,the lungs of the rats showed acute interstitial inflammation,focal edema,hemorrhage and atelectasis.and subcellular damages on the pulmonary blood-barrier.(3)ZAHS,after neutralization in vitro with antiserum against human C3 and C5,lost most of its harmful effects mentioned above.The findings suggest that the harmful effects of ZAHS originate from the fragments of C3 and Cs and they are most effective when its dosage is suitable and there must be a sufficient length of time to react before the phagocytosis of PMNs.The possible mechanism of the origination of the harmful effects of ZAHS is as follows:Before PMNs undertake phagocytosis,they releas the bactericidal and inflamnagenic (O2-and specific granules extra-cellularly.So the intracellular bactericidal activity of PMNs is decreased.The accumulation of PMNs in the pulmonary tissues can damage the PMNs themselves and the adjacent pulmonary blood-barrier,which will leads to the occurrence of infection and multiple organ failure.