Structural characterization of cocktail-like targeting polysaccharides from Ecklonia kurome Okam and their anti-SARS-CoV-2 activities in vitro

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent responsible for the worldwide coronavirus disease 2019 (COVID-19) outbreak. Investigation has confirmed that polysaccharide heparan sulfate can bind to the spike protein and block SARS-CoV-2 infection. Theoretically, similar structure of nature polysaccharides may also have the impact on the virus. Indeed, some marine polysaccharide has been reported to inhibit SARS-Cov-2 infection in vitro, however the convinced targets and mechanism are still vague. By high throughput screening to target 3CLpro enzyme, a key enzyme that plays a pivotal role in the viral replication and transcription using nature polysaccharides library, we discover the mixture polysaccharide 375 from seaweed Ecklonia kurome Okam completely block 3Clpro enzymatic activity (IC50, 0.48 {micro}M). Further, the homogeneous polysaccharide 37502 from the 375 may bind to 3CLpro molecule well (kD value : 4.23 x 10-6). Very interestingly, 37502 also can potently disturb spike protein binding to ACE2 receptor (EC50, 2.01 {micro}M). Importantly, polysaccharide 375 shows good anti-SARS-CoV-2 infection activity in cell culture with EC50 values of 27 nM (99.9% inhibiting rate at the concentration of 20 {micro}g/mL), low toxicity (LD50: 136 mg/Kg on mice). By DEAE ion-exchange chromatography, 37501, 37502 and 37503 polysaccharides are purified from native 375. Bioactivity test show that 37501 and 37503 may impede SARS-Cov-2 infection and virus replication, however their individual impact on the virus is significantly less that of 375. Surprisingly, polysaccharide 37502 has no inhibition effect on SARS-Cov-2. The structure study based on monosaccharide composition, methylation, NMR spectrum analysis suggest that 375 contains guluronic acid, mannuronic acid, mannose, rhamnose, glucouronic acid, galacturonic acid, glucose, galactose, xylose and fucose with ratio of 1.86 : 9.56 : 6.81 : 1.69 : 1.00 : 1.75 : 1.19 : 11.06 : 4.31 : 23.06. However, polysaccharide 37502 is an aginate which composed of mannuronic acid (89.3 %) and guluronic acid (10.7 %), with the molecular weight (Mw) of 27.9 kDa. These results imply that mixture polysaccharides 375 works better than the individual polysaccharide on SARS-Cov-2 may be the cocktail-like polysaccharide synergistic function through targeting multiple key molecules implicated in the virus infection and replication. The results also suggest that 375 may be a potential drug candidate against SARS-CoV-2.

The role of erlotinib in the acute lung injury and the expression of surfactant protein A / 中华急诊医学杂志

Objective To investigate the role of erlotinib in the expression of surfactant protein A (SP-A) in LPS-induced acute lung injury (ALI) of mice model.Methods C57BL/6 mice were randomly (random number)divided into control group (n=6),ER group (n=6),LPS group (n=6),and ER+LPS group (n=6).In the LPS group,2 mg/kg LPS was instilled into trachea of mice to induce lung injury.In control group,normal saline was instilled into trachea of mice instead.In the ER+LPS group and ER group,100 mg/kg of edotinib was instilled into stomach of mice,and one hour later.2 mg/kg LPS was instilled into trachea of mice in ER+PLS group to induce lung injury.Twenty-four hours later,bronchoalveolar lavage fluid (BALF) and lung tissue of mice in four groups were collected.HE staining were used for evaluating pathological changes of lung injury.Lung wet/dry weight ratio,protein concentrations and total cell numbers in the BALF were measured to determine the degree of pulmonary edema.Immunohistochemical staining and Western Blot were used for testing the protein expression of SP-A,Data of multiple groups were analyzed by one way variance (ANOVA) and inter-group comparisons were made by the least significant difference (LSD) tests.Results There was no significant difference in lung injury score (LIS) between control group (0.056±0.008) and ER (0.064±0.037) group,The LIS in LPS group (0.846-±0.047) was higher than that in control group,however the LIS in ER+LPS group (0.279±0.020) was significant lower than that in LPS group (P ＜ 0.05).Lung wet/dry weight,SP-A concentration and total cell numbers in the bronchoalveolar lavage fluid revealed that the degree of pulmonary edema in LPS group was higher than that in control group,and this pulmonary edema was reversed by erlotinib treatment.Immunohistochemical staining and Western blot showed that the expression of SP-A in LPS group was decreased compared with control group,but it was recovered after erlotinib treatment (P ＜ 0.05).Conclusions Erlotinib could protect the LPS-induced ALI,and it may be related to the regulation of SP-A.

Analysis on Drug Resistance of Enterobacteriaceae Bacteria to Carbapenems Antibiotics in Our Hospital / 中国药房

OBJECTIVE:To provide reference for rational use of carbapenem antibiotics in the clinic. METHODS:Enterobac-teriaceae bacteria were collected from our hospital during Jan. 2014-Dec. 2015;semiautomatic microbiological assay instrument was used for strain culture,identification and drug sensitive tests. Modified Hodge test and K-B test were adopted to confirm Klebsiella pneumoniae carbapenemases (KPC)-producing and ESBLs-producing drug resistant strains. RESULTS:During 2014-2015,1 035 strains of Enterobacteriaceae bacteria were detected in our hospital,among which there were 732 strains of Escherichia coli,157 strains of K. pneumonia,136 strains of Enterobacter cloacae and 10 strains of Serratia marcescens. Citrobacter freundii was not found. E. coli and K. pneumonia were highly sensitive to amikacin and carbapenems,but slightly sensitive to most cephalosporin. A total of 64 strains of carbapenems-resistant Enterobacteriaceae bacteria(6.18%)were detected,among which there were 31 strains of carbapenems-resistant E. coli(4.23%),30 strains of carbapenems-resistant K. pneumonia(19.11%),1 strain of carbapenems-re-sistant E. cloacae(0.74%)and 2 strains of carbapenems-resistant S. marcescens(20.00%). The samples were mainly from sputum and urine specimens,which were mainly from neonatal department and ICU. Of 64 drug resistant strains,there were 59 KPC-pro-ducing strains (92.19%) and 3 ESBLs-producing strains (4.69%). CONCLUSIONS:E. coli occupies high proportion among En-terobacteriaceae bacteria,and the number of carbapenems-resistant E. coli and carbapenems-resistant K. pneumoniae is in high lev-el. Drug resistance of Enterobacteriaceae bacteria to carbopenems may be associated with KPC and ESBLs producing. Carbapenem antibiotics should be selected rationally in accordance with medication indications and the results of drug sensitivity test.

HPLC method for determining serum methotrexate concentration in children patients / 国际检验医学杂志

Objective To establish a HPLC method for determining the serum methotrexate(MTX)concentration in children patients for monitoring the blood drug concentration in clinic.Methods The chromatographic analytical column was the Shimaduz C-18 colum(250 mm×4.6 mm,5 μm),the mobile phase was 0.025 mol/L NaH2 PO4 (pH 5.41)-methanol(76∶24,V/V).The flow rate was 1 .0 mL/min.The detection wavelength was 313 nm.The column temperature was set at 35 ℃.The plasma samples were deposited down the protein with 10% HClO4 ,the supernatant liquid 130 μL added with 1 mol/L NaOH 10 μL was directly injected for determination.Results The mass concentration of MTX in the range of 0.00-4.84 μmol/L and 4.84-10.00 μmol/L showed the good linear relation with the peak area,r 1 =1 ,r2 =1 .The mean recovery rates were 61.67%and 71.83%.Intra-day and inter-day precision were <10% and 12%,respectively(n =3).Conclusion The used method is simple,accurate and reliable with high sensitivity,good reproducibility and wide linearity range,which is suitable for the monitoring blood MTX concentration.

Using bone marrow mesenchymal stem cells to treat liver diseases / 中华肝胆外科杂志

Primary hepatocellular carcinoma progresses from liver fibrosis and cirrhosis to eventually result in liver failure and distant metastasis.Surgical resection is the preferred method of treatment for liver cancer while interventional treatment and liver transplantation are the choices to treat end-stage liver cancer.Unfortunately,partial hepatectomy and interventional treatment are not ideal due to the resulting consequence of hepatocyte dysfunction.Extensive clinical application of liver transplants is limited by the lack of available donors and high costs.Over the past decade,researches on bone marrow mesenchymal stem cells (BMSCs)have made remarkable achievements in the medical field.In this review,we summarize the recent progress of BMSCs in the treatment of liver diseases.

Anesthetic action of volatile anesthetics by using Paramecium as a model / 华中科技大学学报（医学）（英德文版）

Although empirically well understood in their clinical administration, volatile anesthetics are not yet well comprehended in their mechanism studies. A major conundrum emerging from these studies is that there is no validated model to assess the presumed candidate sites of the anesthetics. We undertook this study to test the hypothesis that the single-celled Paramecium could be anesthetized and served as a model organism in the study of anesthetics. We assessed the motion of Paramecium cells with Expert Vision system and the chemoresponse of Paramecium cells with T-maze assays in the presence of four different volatile anesthetics, including isoflurane, sevoflurane, enflurane and ether. Each of those volatiles was dissolved in buffers to give drug concentrations equal to 0.8, 1.0, and 1.2 EC50, respectively, in clinical practice. We could see that after application of volatile anesthetics, the swimming of the Paramecium cells was accelerated and then suppressed, or even stopped eventually, and the index of the chemoresponse of the Paramecium cells (denoted as I ( che )) was decreased. All of the above impacts were found in a concentration-dependent fashion. The biphasic effects of the clinical concentrations of volatile anesthetics on Paramecium simulated the situation of high species in anesthesia, and the inhibition of the chemoresponse also indicated anesthetized. In conclusion, the findings in our studies suggested that the single-celled Paramecium could be anesthetized with clinical concentrations of volatile anesthetics and therefore be utilized as a model organism to study the mechanisms of volatile anesthetics.

Anesthetic action of volatile anesthetics by using Paramecium as a model / 华中科技大学学报（医学）（英德文版）

Although empirically well understood in their clinical administration, volatile anesthetics are not yet well comprehended in their mechanism studies. A major conundrum emerging from these studies is that there is no validated model to assess the presumed candidate sites of the anesthetics. We undertook this study to test the hypothesis that the single-celled Paramecium could be anesthetized and served as a model organism in the study of anesthetics. We assessed the motion of Paramecium cells with Expert Vision system and the chemoresponse of Paramecium cells with T-maze assays in the presence of four different volatile anesthetics, including isoflurane, sevoflurane, enflurane and ether. Each of those volatiles was dissolved in buffers to give drug concentrations equal to 0.8, 1.0, and 1.2 EC50, respectively, in clinical practice. We could see that after application of volatile anesthetics, the swimming of the Paramecium cells was accelerated and then suppressed, or even stopped eventually, and the index of the chemoresponse of the Paramecium cells (denoted as I ( che )) was decreased. All of the above impacts were found in a concentration-dependent fashion. The biphasic effects of the clinical concentrations of volatile anesthetics on Paramecium simulated the situation of high species in anesthesia, and the inhibition of the chemoresponse also indicated anesthetized. In conclusion, the findings in our studies suggested that the single-celled Paramecium could be anesthetized with clinical concentrations of volatile anesthetics and therefore be utilized as a model organism to study the mechanisms of volatile anesthetics.

Preventive effect of gastrodin on cognitive decline after cardiac surgery with cardiopulmonary bypass: A double-blind, randomized controlled study / 华中科技大学学报（医学）（英德文版）

Cognitive decline is a common complication after cardiac surgery with cardiopulmonary bypass (CPB), but as such no pharmacological therapy has been shown to be efficacious in preventing the decline. However, gastrodin has been shown to have multi-pharmacological effects on neurological functions. We undertook this study to test the hypothesis that gastrodin would potentially prevent CPB-associated neurocognitive decline. We randomly assigned 200 patients undergoing mitral valve replacement surgery to receive either gastrodin (40 mg/kg) or saline after the induction of anesthesia and subsequently evaluated cognitive function before surgery, at discharge, and at 3rd month after surgery by using a battery of five neurocognitive tests, or adverse effects of gastrodin postoperatively. Neurocognitive decline in postoperative function was defined as a drop of 1 SD or more in the scores on tests of any one of the four domains of cognitive function. Cognitive decline occurred in 9% of the patients in the gastrodin group in contrast to 42% in the control group (P<0.01) at discharge. Cognitive outcome could be determined at 3rd month in 87 patients in the gastrodin group and 89 in the control group. Cognitive decline was detected in 6% in the gastrodin group and 31% in the control group (P<0.01). The incidences of possible adverse effects were similar between two groups. These results indicate that gastrodin is an effective and a safe drug for the prevention of neurocognitive decline in patients undergoing mitral valve replacement surgery with CPB.

Comparison of two techniques for endothacheal intubation in mice / 中国医师杂志

Objective To explore a safe and efficient method for endotracheal intubation in mice.Methods 60 BALB/c mice were random divided into 2 groups, direct intubation under direct vision group, ( n = 30) and direct intubation under light by transillumination group ( n = 30).After successful tracheal intubation at the first judgment, mouse received a tracheal instillation of 3 mg/kg lipopolysaccharide.Bronchoalveolar lavage (BAL) was then performed three times with 0.8 ml sterile saline.Exudate cells were centrifuged and stained with Wright's fluid.Whether the intubation indeed succeeded was judged by the amount of neutrophil infiltrated into the lungs.The mortality after intubation, the time consumed for each successful tracheal intubation and the times of attempting for intubation in two groups were assessed to approve the efficiency of two techniques.Results Eight vs.two mice died within 24 hours in two groups respectively.The time consumed for each successful tracheal intubation and success rate for the first time for two groups were (92.6 ±23.4)s vs (64.0±20.1)s and 53.3% vs 86.7% respectively, which the consumed time was significantly different, while the final success rate almost the same for the two groups.Conclusion Both direct intubation under direct vision and intubation under light by transillumination can successfully intubate the tube into the trachea.But direct intubation under direct vision by transillumination is more efficient and safe in endotracheal intubation in mice.

Inhibition of IL-13-induced collagen I by sIL-13Rα2 in NIH-3T3 fibroblast cells and the profile of sIL-13Rα2/IL-13 in mice with schistosomiasis / 中国人兽共患病学报

To determine the inhibition of IL-13 by recombinant sIL-13Rα2 in NIH-3T3 fibroblast cells for its potential therapeutic value in hepatic fibrosis caused by Schistosoma japanicum in mice . IL-13 and sIL-13Rα2 from liver of BALB/c mice infected with S.japonicum at different infection time (weeks 0,6,8,10 and 12) were analyzed by ELISA and RT-PCR. The recombinant sIL-13Rα2 expression plasmidwas constructed, followed by transfection into NIH-3T3 fibroblast cells. TypeⅠcollagen produced by NIH-3T3 cells were examined by RT-PCR and Western blotting. It was demonstrated that the expression of IL-13 increased gradually after infection, reached peak density (16.1586 pg/mL)at week 8 and then reduced but was still higher than the level of control mice(3.4146 pg/mL;P =0.017 ). The secretion of sIL-13R α2 reached to its peak 10 weeks after infection(4827.426 pg/mL)and then reduced slowly but still higher than normal(4057.112 pg/mL; P=0.021). Meanwhile, the changes in mRNA level of IL-13 and sIL-13R α2 were coincided with that examined by ELISA. Both IL-13 and sIL-13Rα2 reached their peak density (P=0.033) at week 8 and 10 (P=0.025) respectively, and they were followed by a slower degree of decrease. The sIL-13Rα2 could significantly inhibit the effect of IL-13 on NIH-3T3 fibroblast cells, showing decreased mRNA level(P =0.012)and protein level of typeⅠcollagen compared with normal groups(P =0.031). It is concluded that the sIL-13Rα2 can inhibit the effect of IL-13 on NIH-3T3 fibroblast cells which leads to a reduced production of typeⅠcollagen, demonstrating its potential therapeutic value in hepatic fibrosis of schistosomiasis.

The protective effect of propofol on erythrocytes during cardiopulmonary bypass / 华中科技大学学报（医学）（英德文版）

To evaluate the relationship between erythrocyte injury and intracellular calcium ion overload, and the protective effect of propofol on erythrocytes during cardiopulmonary bypass (CPB). 40 children with congenital heart diseases who underwent surgical repair under CPB were included. The patients were randomly divided into two groups: control group (group C) and propofol group (group P). Anesthesia was maintained in the patients with 6 mg/kg/h propofol in Group P, and those in the Group C inhaled 1%-2% isoflurane. The blood samples were taken before CPB, 30 min after CPB, at the end of CPB, and 2 h and 24 h after CPB to measure the content of erythrocyte intracellular calcium ion (E-Ca2+), Ca2+-Mg2+-ATPase and Na+-K+-ATPase activities, index filtration of erythrocytes (IF), mean corpuscular volume (MCV) and the concentration of plasma free hemoglobin (F-Hb). Results showed that in the control group, E-Ca2+, IF, MCV and F-Hb were gradually increased and Ca2+-Mg2+-ATPase and Na+-K+-ATPase activities were decreased. The increase of E-Ca2+ was linearly paralleled to IF, MCV and F-Hb. In propofol group, all the above-mentioned parameters were significantly improved (P<0.05). This study suggests that erythrocyte injury is related to elevation of intracellular calcium during CPB and propofol has a protective effect on erythrocyte injury.

Therapeutic effect of propofol in the treatment of endotoxin-induced shock in rats / 华中科技大学学报（医学）（英德文版）

To assess the potential therapeutic effect of propofol in the treatment of endotoxemia, 76 rats were randomly assigned to 5 groups: control group(A), endotoxemic group(B), pre-treatment group(C), simultaneous treatment group(D) and post-treatment group(E). Five h after endotoxin injection, PO2, pH, MAP, plasma concentrations of Nitrite/nitrate (NO2-/NO3-) and mortality rates were assessed in each group. After the rats were sacrificed, lung tissue was sampled to measure myeloperoxidase (MPO) activity and tumor necrosis factor (TNF)-alpha contents. It was found that endotoxin injection produced progressive hypotension, metabolic acidosis, and a large increase in the plasma NO2-/NO3- concentrations and increased mortality rates in 5 h. Endotoxin injection significantly increased MPO activity and TNF-alpha contents in lung tissue (P < 0.01 or P < 0.05). These changes response to endotoxin were significantly attenuated in the groups B, C and D. But these beneficial effects were blunted in the group E. The results suggest that propofol administration may offer advantages in endotoxemia.

Effects of ropivacaine and bupivacaine on rabbit myocardial energetic metabolism and mitochondria oxidation / 华中科技大学学报（医学）（英德文版）

To compare the cardiotoxicity induced by ropivacaine and bupivacaine and to investigate the mechanism of cardiotoxicity, 24 mature New Zealand rabbits were divided randomly into control group (group C), ropivacaine group (group R) and bupivacaine group (group B). Hearts were drawn out rapidly from the anesthetized animals and cardiac perfusion was performed immediately. Ropivacaine 500 ng/ml (group R) or bupivacaine 500 ng/ml (group B) was added to the perfusion solution. Ventricular myocardial ATP, ADP and AMP were measured with high performance liquid chromatogram. The ability of myocardial mitochondria oxidation to pyruvate or palmitoylcarnitine was detected with Clark electrode. Our results showed that myocardial ATP and ADP decreased significantly (P < 0.05) in group R and most significantly (P < 0.01) in group B as compared with group C. Myocardial ATP and ADP decreased most significantly (P < 0.01) in group B as compared with group R. The changes of myocardial AMP revealed significant difference among three groups. The changes of pyruvate oxidation exhibited no significant difference among the three groups. Palmitoylcarnitine oxidation decreased markedly (P < 0.05) in group R and most significantly (P < 0.01) in group B as compared with group C. The present study indicated that the inhibition of lipid substrate oxidation may be responsible for the cardiotoxicity induced by bupivacaine and ropivacaine. The cardiotoxicity induced by ropivacaine is far more less than bupivacaine.

The Protective Effect of Propofol on Erythrocytes during Cardiopulmonary Bypass / 华中科技大学学报（医学）（英德文版）

To evaluate the relationship between erythrocyte injury and intracellular calcium ion overload, and the protective effect of propofol on erythrocytes during cardiopulmonary bypass (CPB), 40 children with congenital heart diseases who underwent surgical repair under CPB were studied. The patients were randomly divided into two groups: control group (group C) and propofol group (group P). Anesthesia was maintained in the patients in group P with 6 mg*kg-1*h-1 propofol, and those in the group C inhaled 1 %-2 % isoflurane. The blood samples were taken before CPB, at the 30th min of CPB, at the end of CPB, and 2 h and 24 h after CPB to measure the content of erythrocyte intracellular calcium ion (E-Ca2+), Ca2+-Mg2+-ATPase and Na+-K+-ATPase activities, index filtration of erythrocytes (IF), mean corpuscular volume (MCV) and the concentration of plasma free hemoglobin (F-HB). Results showed that in the control group, E-Ca2+, IF, MCV and F-Hb were gradually increased and Ca2+-Mg2+-ATPase and Na+-K+-ATPase activities were decreased. The increase of E-Ca2+ was linearly paralleled to IF, MCV and F-Hb. In propofol group, all the above-mentioned parameters were significantly improved (P<0.05). This study suggests that erythrocyte injury is related to elevation of intracellular calcium during CPB and propofol has a protective effect on erythrocyte injury.

The Protective Effect of Propofol on Erythrocytes during Cardiopulmonary Bypass / 华中科技大学学报（医学）（英德文版）

To evaluate the relationship between erythrocyte injury and intracellular calcium ion overload, and the protective effect of propofol on erythrocytes during cardiopulmonary bypass (CPB), 40 children with congenital heart diseases who underwent surgical repair under CPB were studied. The patients were randomly divided into two groups: control group (group C) and propofol group (group P). Anesthesia was maintained in the patients in group P with 6 mg*kg-1*h-1 propofol, and those in the group C inhaled 1 %-2 % isoflurane. The blood samples were taken before CPB, at the 30th min of CPB, at the end of CPB, and 2 h and 24 h after CPB to measure the content of erythrocyte intracellular calcium ion (E-Ca2+), Ca2+-Mg2+-ATPase and Na+-K+-ATPase activities, index filtration of erythrocytes (IF), mean corpuscular volume (MCV) and the concentration of plasma free hemoglobin (F-HB). Results showed that in the control group, E-Ca2+, IF, MCV and F-Hb were gradually increased and Ca2+-Mg2+-ATPase and Na+-K+-ATPase activities were decreased. The increase of E-Ca2+ was linearly paralleled to IF, MCV and F-Hb. In propofol group, all the above-mentioned parameters were significantly improved (P<0.05). This study suggests that erythrocyte injury is related to elevation of intracellular calcium during CPB and propofol has a protective effect on erythrocyte injury.

Peroxisome proliferator-activated receptor ? agonist attenuates ICAM-1 and CINC-1 expression in lungs of rats with acute lung injury / 中国病理生理杂志

AIM: To investigate the effects of rosiglitazone(ROSI),an agonist of peroxisome proliferator-activated receptor ?(PPAR?),on the lung expression of intercellular adhesion molecule-1(ICAM-1) and cytokine-induced neutrophil chemoattractant(CINC) in rats with acute lung injury. METHODS: Thirty-six male Wistar rats were randomly divided into six groups: control group,ROSI group,GW9662(a PPAR? antagonist) group,lipopolysaccharide(LPS,6 mg/kg,iv) group,ROSI-LPS group(0.3 mg/kg ROSI iv 30 min prior to LPS) and GW9662-ROSI-LPS group(0.3 mg/kg GW9662,iv,20 min before ROSI).Four hours after LPS injection,wet/dry weight(W/D) ratio,myeloperoxidase (MPO) activity,malondialdehyde(MDA) and CINC-1 concentrations were assayed in the lung tissues.Immunohistochemical analysis of ICAM-1 expression was also studied.RESULTS: Pretreatment with ROSI significantly attenuated LPS-induced increases in W/D ratio,MPO activity,MDA and CINC-1 concentrations as well as ICAM-1 expression in the lung tissues.The specific PPAR? antagonist GW9662 antagonized the effects of ROSI.CONCLUSION: Pretreatment with ROSI reduces LPS-induced lung injury in rats.The mechanism involves inhibition of the lung expression of ICAM-1 and CINC-1 by the activation of PPAR?.

Effects of ropivacaine and bupivacaine on energy supply and mitochondrial oxidation of isolated rabbit heart muscle / 中华麻醉学杂志

Objective The purpose of this study was to investigate the effects of ropivacaine and bupivacaine on energy supply and mitochondrial oxidation phosphorylation of isolated rabbit heart muscle in order to compare the cardiotoxicity of the two local anestheties and explain the mechanism Methods Twenty four New Zealand rabbits weighing (2135?345)g were randomly divided into three groups of 8 animals each: control group(C); ropivacaine group(R) and bupivacaine group (B) The animals were anesthetized with intraperitoneal urethane 2g?kg -1 and heparinized (heparin 4mg?kg -1 ) Heart was quickly removed and connected to Langendoff preparation and perfused with oxygenated Krebs Henseleit buffer solution at normothermia(37℃) via aorta After 15 min perfusion, ropivacaine (group R) or bupivacaine(group B) was added to perfusate(500ng?ml -1 ) The heart was then perfused for another 10 min Myocardial tissue was taken from ventricle for determination of ATP, ADP and AMP content with high performance liquid chromatography Myocardial mitochondria was prepared and its oxidation of pyruvate and palmitoylcarnitine was measured using Clark electrode Results Myocardial ATP and ADP decreased significantly in group R and B as compared with those in group C The decrease in ATP and ADP in group B was more than that in group R (P

The mechanism of erythrocyte injury during cardiopulmonary bypass in children and the protective effect of propofol / 中华麻醉学杂志

Objective To evaluate the relationship between erythrocyte injury and intracellular calcium ion overload and the protective effect of propoful on erythrocytes during cardiopulmonary bypass(CPB) Methods Thirty children (male 21,female 9) with congential heart disease scheduled for elective open heart surgery with CPB were randomly divided into two groups : control group (C) and propoful group (P) The congential heart disease included ventricular septal defect (17 patients) and atrial septal defect (13 patients) The age ranged from 2 10 yr and body weight 10 35kg Anesthesia was induced with fentanyl 20 ?g/kg and vecuronime 0 1mg/kg in both groups and maintained with low concentration of isoflurane inhalation in group C and propofol intravenous infusion at a rate of 6mg?kg -1 ?h -1 in group P Blood samples were taken before and 20 min after CPB was began, at the end of CPB and 2 and 4h after CPB for determination of intracellular calcium ion content (E Ca 2+ ), Ca 2+ ,Mg 2+ ATPase and Na +, K + ATPase activities, erythrocyte filtration index(IF), mean corpuscular volume (MCV) and plasma free hemoglobin concentration (F Hb) Results In control group E Ca 2+ ,IF,MCV and F Hb gradually increased and Ca 2+ ,Mg 2+ ATPase and Na +,K + ATPase activities decreased during CPB The increase in E Ca 2+ was linearly paralleled to IF,MCV and F Hb There was no significant change in all the above mentioned parameters The difference in these parameters was significant between the two groups Conclusions The study suggests that erythrocyte injury is related to the increase in intracellular calcium ion during CPB and propoful has a protective effect on erythrocyte against injury due to its abilities to scavenge free radical and inhibit calcium channel

Effects of cardiopuimonary bypass on platelet counts and platelet release reaction / 中华麻醉学杂志

To investigate the effects of cardiopulmonary bypass (CPB)on the aquired platelet damage and the relationship between the aquired platelet damage and non-surgical postoperative bleeding. Method: Platelet counts (BPC), alphagranule membrane protein (GMP-140), ?-thromboglobulin (?-TG), platelet factor 4 (PF_4), and 5-hydrooxytryptamine levels were measured in 20 patients undergoing cardiac surgery before CPB, 30 min during CPB, 10 min after CPB, and 2, 12, and 24 hours postoperatively. The numbers of patients with bleeding volume over 200 ml within 24 hours postoperatively were counted. Result: BPC decreased markedly during CPB, but never decreased to the degree of 50?10~9/L. GMP-140, ?-TG, PF_4 and 5-HT levels were significantly increased during CPB until 12 hours postoperatively. Eight patients(40%) got the bleeding volume over 200 ml within 24 hours postoperatively. Conclusion: Platelet release reaction is violent during cardiac surgery with CPB. A large number of platelets dysfunctioned because of granula releasing or damage may be the main cause of non-surgical postoperative bleeding.

Antioxidant potential of propofol during cardiopulmonary bypass in adults / 中华麻醉学杂志

Objective To investigate the antioxidative potential of propofol in adults undergoing open-heart surgery with cardiopulmonary bypass (CPB) Methods Thirty patients scheduled for cardiac procedures with CPB were selected Propofol (01 mg?kg -1?min -1, group A) or fentanyl (5?g?kg -1?min -1, group B) was administered to maintain anesthesia after aorta was cross-clamped Blood samples were drawn pre-anesthesia, pre-CPB, 30 min following CPB, at the end of CPB, 1h after CPB, at the end of operation, 12 and 24 h postoperatively RBC suspension was prepared to measure the erythrocyte glucose-6-phosphate dehydrogenase (G-6-PD) and phosphofructokinase (PFK) activities, total erythrocyte reduced glutathione (GSH) and oxidized GSH (GSSG) and GSH/GSSG ratio was calculatedResults In group A, G-6-PD and PFK activities and GSH/GSSG ratio remained unchanged significantly during CPB and postoperatively In group B, G-6-PD activity increased and PFK activity and GSH/GSSG ratio decreased significantly from 30th min of CPB to 12th h postoperativelyConclusions Propofol attenuates free radical activity during CPB to reduce the free radical-induced injury, while fentanyl has no effect on free radical reduction during CPB