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Objective:To evaluate the relationship between intestinal mucosal mast cells and intestinal flora during intestinal ischemia-reperfusion(I/R) in mice.Methods:Forty-eight healthy male C57BL/6 mice, aged 9-12 weeks, weighing 20-25 g, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (group Sham), sham operation plus mast cell membrane stabilizer cromolyn sodium (CS) group (group Sham+ CS), I/R group and I/R+ CS group.The intestinal I/R injury was induced by clamping the superior mesenteric artery for 45 min followed by 4 h of reperfusion in I/R and I/R+ CS groups.The superior mesenteric artery was only exposed but not occluded in Sham and Sham+ CS groups.CS 50 mg/kg was intraperitoneally injected once a day for two weeks before surgery in Sham+ CS and I/R+ CS groups.The equal volume of normal saline was intraperitoneally injected once a day for two weeks before surgery in Sham and I/R groups.Mice were then sacrificed and intestinal tissues were harvested for examination of the pathological changes of intestinal mucosa with a light microscope after HE staining, and the damage to intestinal tissues was assessed and scored according to Chiu.The expression of mast cell tryptase was detected using the immunohistochemical SP staining method, and the mast cells were counted.The intestinal contents were collected, the total amount of bacteria in intestinal flora was detected by 1% agarose gel electrophoresis, and the diversity (Chao1 index and Shannon index) and relative abundance of intestinal microbial bacteria were detected by 16S rDNA sequencing method. Results:Compared with Sham group, the Chiu′s score and mast cell count were significantly increased, the expression of mast cell tryptase was up-regulated, the total amount of intestinal flora bacteria was increased, Chao1 index and Shannon index were decreased, the relative abundance of Bacteroidetes, Proteobacteria, Bacteroides and Enterobacteriaceae were increased, and the relative abundance of Firmicutes, Lactobacillus and Bifidobacteria was reduced in I/R group ( P<0.05 or 0.01), and no significant change was found in the parameters mentioned above in Sham+ CS group ( P>0.05). Compared with I/R group, the Chiu′s score and mast cell count were significantly decreased, the expression of mast cell tryptase was down-regulated, the total amount of intestinal flora bacteria was decreased, Chao1 index and Shannon index were increased, the relative abundance of Bacteroidetes, Proteobacteria, Bacteroides and Enterobacteriaceae were reduced, and the relative abundance of Firmicutes, Lactobacillus and Bifidobacteria was increased in I/R+ CS group ( P<0.05 or 0.01). Conclusion:The activation of intestinal mucosal mast cell can lead to imbalance of intestinal flora, decrease the number of probiotics and increase the number of potential pathogenic bacteria, and thus be involved in the pathophysiological mechanism of intestinal I/R injury in mice.
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Objective To evaluate the role of tryptase in intestinal ischemia-reperfusion (Ⅰ/R) injury in rats and the relationship with inflammatory responses,lipid peroxidation and cell apoptosis.Methods Thirty clean-grade healthy male Sprague-Dawley rats,aged 2-3 months,weighing 150-270 g,were divided into 3 groups (n =10 each) using a random number table method:sham operation group (Sham group),intestinal Ⅰ/R group (Ⅱ/R group) and tryptase inhibitor group (PRTM group).Intestinal Ⅰ/R was produced by occlusion of the superior mesenteric artery for 75 min followed by 4-h reperfusion in Ⅱ/R and PRTM groups.The superior mesenteric artery was only exposed but not occluded in Sham group.Protamine 2.5 mg/kg was injected via the caudal vein at 5 min of before reperfusion in PRTM group.The rats were sacrificed at the end of reperfusion,and the small intestinal tissues 1 cm in length 5 cm away from the terminal ileum were removed for examination of the pathological changes (with a light microscope) and for determination of intestinal mucosal mast cell (IMMC) count and expression of tryptase in mast cells (by immunohistochemical SP staining),malondialdehyde (MDA) content (using thiobarbituric acid assay),superoxide dismutase (SOD) activity (by xanthine oxidase method),expression of caspase-3 (by Western blot) and contents of tumor necrosis factor alpha (TNF-ot) and interleukin-6 (IL-6) (by enzyme-linked immunosorbent assay).The degree of intestinal tissue damage was graded using Chiu's scoring system.Results Compared with Sham group,the pathological changes were significantly accentuated,Chiu's score was increased,the expression of tryptase was up-regulated,the IMMC count and contents of TNF-α,IL-6 and MDA were increased,the activity of SOD was decreased,and the expression of caspase-3 was up-regulated in Ⅱ/R and PRTM groups (P＜0.05).Compared with Ⅱ/R group,the pathological changes were significantly attenuated,Chiu's score was decreased,the expression of tryptase was down-regulated,the IMMC count and contents of TNF-α,IL-6 and MDA were decreased,the activity of SOD was increased,and the expression of caspase-3 was down-regulated in PRTM group (P＜0.05).Conclusion Tryptase is involved in the process of intestinal Ⅰ/R injury though promoting inflammatory responses,lipid peroxidation and cell apoptosis in rats.
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Objective To evaluate the changes in the expression of small intestinal thioredoxin 2 (Trx2) during different periods after orthotopic liver autotransplantation (OLAT) in rats.Methods Forty Sprague-Dawley rats,aged 8-10 weeks,weighing 210-260 g,were randomly divided into 2 groups using a random number table:sham operation group (group S,n =8) and OLAT group (n =32).Intestinal tissues were removed at 4,8,16 and 24 h after OLAT for microscopic examination and for determination of the levels of superoxide anion (O2--),hydrogen peroxide (H2 O2),glutathione peroxidase (GSH-Px),reduced glutathione (GSH) and Trx2.Intestinal damage was assessed and scored according to Chiu.Results Compared with S group,the Chiu's score and O2--activity at 4,8 and 16 h after OLAT and H2O2 content at 4 and 8 h after OLAT were significantly increased,and the levels of GSH-Px and GSH and expression of Trx2 at 4 and 8 h after OLAT were decreased in OLAT group (P ＜ 0.05).Chiu' s score at 4,16 and 24 h after OLAT and H2O2 content at 16 and 24 h after OLAT were significantly lower than those at 8 h in OLAT group (P ＜ 0.05).Conclusion The rats undergo decreased antioxidant capacity in the early phase and recovery in the late phase mediated by small intestinal Trx2 after OLAT.
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Objective To evaluate the effect of lipo-alprostadil on lung injury in patients undergoing orthotopic liver transplantation.Methods Forty-eight ASA Ⅱ-Ⅳ patients of both sexes,aged 45-64 yr,weighing 45-70 kg,scheduled for elective orthotopic liver transplantation,were randomly assigned to one of 2 groups (n =24 each):control group (group C) and lipo-alprostadil group (group A).Anesthesia was induced with midazolam,propofol,fentanyl and vecuronium and maintained with sevoflurane,sufentanil and vecuronium.The patients were tracheal intubated and mechanically ventilated.Lipo-alprostadil 5 μg in 10 ml of normal saline was infused intravenously and slowly over 30 min before induction of anesthesia and at 1 h of neohepatic phase in group A.Lipoalprostadil was not administrated in group C.Peak inspiratory pressure (PIP),mean inspiratory pressure (Pmean),dynamic lung compliance (Cd),oxygenation index (OI),respiratory index (RI) and the concentrations of inflammatory cytokines in exhaled breath condensate (EBC) were recorded immediately before operation,at the end of operation,and at 24 h after operation.The occurrence of pulmonary complications was recorded within 7days after operation.Results Compared with group C,PIP,Pmean,RI,and TNF-α and IL-8 concentrations in EBC were significantly decreased,while Cd and OI were increased at the end of operation and 24 h after operation,and the incidence of acute lung injury and pulmonary infection were decreased within 7 days after operation (P ＜0.05),and no significant change in the other indexes was found in group A (P ＞ 0.05).Conclusion Lipo-alprostadil has protective effect on lung in patients undergoing orthotopic liver transplantation.
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Objective To evaluate the cardiac function of the patients with liver cirrhosis before orthotopic liver transplantation(OLT)using Swan-Ganz catheter.Methods Sixty ASAⅡ-Ⅳ patients aged 45-64 yr with liver cirrhosis scheduled for OLT without veno-venous bypass were allocated into 2 groups according to preoperative liver function:compensated group(group C,n=28)and decompensated group(group H,n=32).Anesthesia was induced with midazolam 3-5 mg,fentanyl 0.15-0.2 mg,propofol 1 mg/kg and vecuronium 0.1 mg/ks and maintained with 0.5%-3.0% isoflurane,fentanyl 0.05-0.10 mg and vecuronium 4 mg/h.The patients were mechanically ventilated after tracheal intubation,and P_(ET)CO_2 was maintained at 30-45 mm Hg.Radial artery was cannulated and Swan-Ganz catheter was placed via right internal jugular vein for monitoring of mean arterial pressure(MAP),cardiac output(co),cardiac index(CI),right ventricular ejection fraction(RVEF),mean pulmonary arterial pressure(MPAP),pulmonary arterial wedge pressure(PAWP),right atrial pressure(RAP),right ventricular end-diastolic volume(RVEDV),fight ventricular end-systolic volume(RVESV)and stroke volume index(SVI).Right and left ventricular stroke work index(RVSWI,LVSWI)and systolic and pulmonary vascular resistance(SVR,PVR)were calculated.Results CO,CI,SVI,MPAP,PAWP,RVEDV,RVESV,RVSWI and LVSWI were significantly elevated in group H as compared with group C indicating hyper-hemodynamic state.The SVR and PVR were significantly decreased in group H.There was no significant difference in HR,MAP,RAP and RVEF between the two groups.Conclusion The patients with decompensated liver function before OLT are in a hyper-hemodynamic state.More attention should be paid to perioperative myocardial protection.
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OBJECTIVE To explore the application of hydrogen dioxide hypothermic plasma sterilizing system(STERRAD 100S) in operating room,and study the cost.METHODS The cost of sterilizing endoscopic instruments in operating room was analyzed.And the method of using medical material was improved as follows: use the non-woven fabrics and transparent packing bags repeatly to increase the utilization efficiency of medical material.RESULTS Compared with the traditional method,the cost of sterilizing instruments by STERRAD 100S decreased 2-3 times,which brought much economic benefit.CONCLUSIONS 100S Can improve the working efficiency,reduce inventory level of endoscopic instruments and sterilizing cost,which can relief patient economic burden and ensure patient interest.
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Objective To evaluate the perioperative monitoring of adult patients with hepatic encephalopathy under orthotopic liver transplatation (OLT).Methods Combined intravenous and inhalational general anesthesia was used for 18 patients with hepatic encephalopathy from October 2003 to August 2004. Rapid-sequence induction was performed. Propofol, 1 to 1.5 mg/kg, and fentanyl, 4 ?g/kg, were used. Norcuron, 0.1 mg/kg, was added to facilitate tracheal intubation. All patients were subjected to piggyback liver transplantation. Hemodynamics, respiratory, blood gas, blood biochemistry, coagulation function, body temperature, liver and kidney functions, urine output, and bleeding output were monitored during operation. According to the situations of patients, platelet, cryoprecipitate, fibrinogen, coagulation factors and ulinastatin were administrated.Results Eighteen patients tolerated the operation. Only 4 patients died of multiple organ dysfunction syndrome after operation. The survival rate reached was up to 77.8 %. The main blood gas change during perioperative phase was metabolic acidosis and hyposodium, hypokaleamia, hypocalcium. The main hemodynamics change during operation was that HR was increased significantly, and CO was higher than normal value before operation, and CO, CVP, SPAP and DPAP decreased significantly in anhepatic stage. Compared with those before operation, ALT, AST, BUN and Cr were increased significantly in neohepatic stage.Conclusions It is very important to pay more attention to these patients with hepatic encephalopathy during different stages of OLT. Drugs not affecting the function of liver and kidney should be selected. Benzodiazepine should be avoided. Supplementation of coagulation factors, CRBC and electrolyte was necessary. The key point is to protect renal function, maintain enough urine output and treat brain edema.
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AIM: To study the changes of serum levels of nitric oxide (NO) and nitric oxide synthase (NOS) in patients during liver transplantation. METHODS: Samples were obtained from 30 patients in end liver disease at five time points during liver transplantation. NO level and NOS activity were measured by radioimmunoassay and colorimetry, respectively. Arterial and mixed venous blood samples used for blood gas analysis were taken at the same time. Intrapulmonary shunt (Qs/Qt) was calculated according to the standard formula. The hemodynamics parameters including continuous cardic output (CO), HR, MABP, CVP, SVR were measured during liver transplantation. RESULTS: (1) NO_2-/NO_3-level at 10 min before anhepatic period was significantly higher than the baseline level. Compared with NO_2-/NO_3-level at 10 min before anhepatic period, NO_2-/NO_3-level at 30 min after anhepatic period was significantly decreased. NO_2-/NO_3-level at 30 min after neohepatic period was significantly higher than the baseline level and at 30 min after anhepatic period. (2) No significant change of tNOS activity was observed. Compared with the baseline activity of inducible nitric oxide synthase (iNOS), the activity at 10 min before anhepatic period and at 30 min after neohepatic period was significantly increased. The activity at 30 min after neohepatic period was significantly higher than that at 30 min after anhepatic period. (3) MABP decreased significantly when opening the inferior vena cava. CO and CVP decreased in the anhepatic stage and increased in the reperfusion stage. SVR increased during anhepatic stage and decreased significantly during neohepatic period. (4) Qs/Qt decreased significantly during anhepatic stage and increased significantly at 30 min after neohepatic period. CONCLUSIONS: Serum level of NO and NOS activity are significantly changed during liver transplantation. High level of NO may result in low systemic vascular resistance and increasing in intrapulmonary shunt.