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.

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.

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

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

Objective: To evaluate the safety of controlled hypotension produced by human ?-calcitonin gene-related peptide(?-hCGRP)and sodium nitroprusside. Method: Twenty SD rats were randomly divided into ?-hCGRP(C) and sodium nitroprusside(S) groups. Animals in group C were infused intravenously with 0.001％ ?-hCGRP and those in group S with 0.01％ sodium nitroprusside. The mean blood pressure of rat was reduced to 6.7kPa and maintained for 30 min. Cerebral glucose,lactate,ATP, and phosphocreatine levels were measured at 0 and 30th min of hypotension. Result: Cerebral glucose level increased slightly,lactate level decreased significantly,ATP and phosphocreatine contents did not change in group C at 30th min of hypotension compared with those at 0 min of hypotension,respectively.Cerebral glucose level increased slightly,lactate concentration increased significantly,ATP and phosphocreatine contents decreased significantly in group S at 30th min of hypotension in comparison with those at 0 min of hypotension, respectively. Conclusion: Hypotension induced with alpha-hCGRP can maintain adequate cerebral oxygen supply,but with sodium nitroprusside may cause cerebral hypoxia.

Objective: Effects of human ?-calcitonin gene-related peptide (?-hCGRP) on renal function and renin-angiotensin system were studied during controlled hypotension. Method:Twenty SD rats were divided into ?-hCGRP (C) and sodium nitroprusside (S) groups. The mean blood pressure of rats was reduced to and maintained at 6.7kPa for 1h. The glomerular filtration rate (GFR),effective renal plasma flow (ERPF),plasma renal activity (PRA),angiotensin Ⅱ(AⅡ), urine volume, heart rate (HR)and the excretory rates of sodium,chloride and potassium of rats were measured beforeand 1h following hypotension. Result:GFR and ERPF increased significantly at the end of 1h hypotension when compared to pre-hypotension in the group C. GFR and ERPF decreased mark during hypotension period in group S. In both groups during hypotension,PRA and A Ⅱ increased by eight-folds or so,the excretory rates of sodium and chloride decreased significantly and the excretory rate of potassium increased slightly, urine volume de creased and HR increased markly. Conclusion:The ?-hCGRP,a potent vasodilator,has a protective effect on renal function as a controlled hypotensive agent. Sodium nitroprusside may harm renal function. The increased excitation of renin-angiotensin system may be responsible for the changes of urine volume,heart rate and the excretory rates of sodium ,chloride and potassium during hypotension.

To evaluate the influence of human ?-caleitonin gene related peptide (?-hCGRP) used in the controlled hypotension on hepatic blood flow, oxygen supply and oxygen consumption in rats. Method: Alpha-hCGRP and sodium nitroprusside were administered intravenously to reduce the blood pressure of rats to 6.7 kPa maintained for 1 h. Radiomierospheres were employd to measure hepatic arterial blood flow (HABF), portal venous blood flow (PVBF), and total hepatic blood flow (TABF). The arterial0portal,and hepatic venous blood gases were analyzed. The hepatic oxygen supply and oxygen consumption were calculated according to values of blood flow and gases. Result: HABF, TABF, and hepatic oxygen supply increased significantly, and PVBF decreased slightly during ?-hCGRPinduced hypotension. HABF, TABF and hepatic oxygen consumption increased significantly, and hepatic oxygen supply decreased significantly during sodium nitroprusside hypotension. Rebound hypertension occurred after the discontinua tion of sodium nitroprusside rather than ?-hCGRP. Conclusion: Alpha-hCGRP does not cause hepatic hypoxia during controlled hypotension, while sodium nitroprusside may result in hepatic hypoxia. Alpha hCGRP is more adequate to be used to controlled hypotension than sodium nitroprusside. No rebound hypotension is another advantage of ?-hCGRP.

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

Abstract: Objective To investigate the preventive and therapeutic effects of anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin on neonatal mice infected with human rotavirus (HRV), and to analyze the underlying mechanism. Methods Hybridoma technology was used to prepare Ab2β of lactadherin. One hundred and twenty 7-day-old Kunming mice were randomly divided into groups A, B, C and control, each consisting of 30 mice. Groups A, B, and C were all infected with HRV via oral gavage. Group A received no treatment, group B was orally administered lactadherin for 7 days prior to infection, and group C was orally administered lactadherin for 7 days after infection, the control group was orally administered cell culture medium that did not contain the virus. The clinical manifestations (diarrhea, body weight) at different time points after infection of the neonatal mice in each group were observed, and the content of rotavirus antigen in the feces of neonatal mice was detected by enzyme-linked immunosorbent assay (ELISA). After HRV infection for 7 days, immunohistochemical staining was used to examine the expression level of intercellular adhesion molecule-1 (ICAM-1) in mouse small intestinal tissues in each group. Results No diarrhea occurred in the control group at any time point. Groups A, B, and C showed diarrhea symptoms after HRV challenge for 1 day. The degree of diarrhea in groups B and C was lower than that in group A at 2-4 days after HRV challenge, and the difference was statistically significant (P<0.05). The HRV antigen content in the feces of the neonatal mice in groups B and C was lower than that in group A at 1-5 days after HRV challenge, and the difference was statistically significant (P<0.05). There was no significant difference in the degree of diarrhea and HRV antigen content between groups B and C at each time point (P>0.05). There was no significant difference in the body weight of the neonatal mice in each group before infection and 1 day after infection (P>0.05); the weight of neonatal mice in groups B and C was higher than that in group A at 3, 5 and 7 days after HRV challenge, and the difference was statistically significant (P<0.05), and there was no significant difference in body weight between groups B and C at each time point after HRV challenge (P>0.05). The number of ICAM-1 expressing cells in the small intestine of the three groups A, B, and C was higher than that of the control group after HRV challenge for 7 days, and the difference was statistically significant (P<0.05). The cell number and gray value of ICAM-1 expressing cells in groups B and C were lower than those in group A, and the difference was statistically significant (P<0.05). Conclusions Anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin has a good preventive and therapeutic effects on human rotavirus infection in neonatal mice, and can significantly improve diarrhea symptoms and reduce HRV viral load. Its specific mechanism may be related to the inhibition of ICAM-1.

Abstract: Objective To investigate the preventive and therapeutic effects of anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin on neonatal mice infected with human rotavirus (HRV), and to analyze the underlying mechanism. Methods Hybridoma technology was used to prepare Ab2β of lactadherin. One hundred and twenty 7-day-old Kunming mice were randomly divided into groups A, B, C and control, each consisting of 30 mice. Groups A, B, and C were all infected with HRV via oral gavage. Group A received no treatment, group B was orally administered lactadherin for 7 days prior to infection, and group C was orally administered lactadherin for 7 days after infection, the control group was orally administered cell culture medium that did not contain the virus. The clinical manifestations (diarrhea, body weight) at different time points after infection of the neonatal mice in each group were observed, and the content of rotavirus antigen in the feces of neonatal mice was detected by enzyme-linked immunosorbent assay (ELISA). After HRV infection for 7 days, immunohistochemical staining was used to examine the expression level of intercellular adhesion molecule-1 (ICAM-1) in mouse small intestinal tissues in each group. Results No diarrhea occurred in the control group at any time point. Groups A, B, and C showed diarrhea symptoms after HRV challenge for 1 day. The degree of diarrhea in groups B and C was lower than that in group A at 2-4 days after HRV challenge, and the difference was statistically significant (P<0.05). The HRV antigen content in the feces of the neonatal mice in groups B and C was lower than that in group A at 1-5 days after HRV challenge, and the difference was statistically significant (P<0.05). There was no significant difference in the degree of diarrhea and HRV antigen content between groups B and C at each time point (P>0.05). There was no significant difference in the body weight of the neonatal mice in each group before infection and 1 day after infection (P>0.05); the weight of neonatal mice in groups B and C was higher than that in group A at 3, 5 and 7 days after HRV challenge, and the difference was statistically significant (P<0.05), and there was no significant difference in body weight between groups B and C at each time point after HRV challenge (P>0.05). The number of ICAM-1 expressing cells in the small intestine of the three groups A, B, and C was higher than that of the control group after HRV challenge for 7 days, and the difference was statistically significant (P<0.05). The cell number and gray value of ICAM-1 expressing cells in groups B and C were lower than those in group A, and the difference was statistically significant (P<0.05). Conclusions Anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin has a good preventive and therapeutic effects on human rotavirus infection in neonatal mice, and can significantly improve diarrhea symptoms and reduce HRV viral load. Its specific mechanism may be related to the inhibition of ICAM-1.