Objective To investigate the effect of cold autologous blood cardioplegia and HTK solution on changes of superoxide dismutase (SOD) and Matrix metalloproteinase-2 (MMP-2) in infant coronary sinus vein blood,which underwent ventricular septal defect suture in extracorporeal circulation,in order to reveal their protective effect of autologous cold blood cardioplegia on immature myocardium.Methods Sixty cases with ventricular septal defect,aged less 1-year-old,were randomly divided into experimental group (n =30) and control group(n =30).Autologous cold blood cardioplegia was obtained on aortic blood before aortic root draw in extracorporeal circulation with K + concentration of 20 mmol/L and 4 ℃ temperature.The experimental group was used cold autologous blood cardioplegia as arresting and protecting cardioplegia,while the control group used HTK solution.Blood sample from coronary sinus and vein was obtained at immediately before aortic cross-clamping,1 minute and 15 minutes after aortic cross-opening.SOD and MMP-2 levels were determined by enzyme-Linked Immuno Sorbent Assay (ELISA).Results The levels of SOD and MMP-2 levels within the group,between the groups and interaction were significant difference (P ＜ 0.05).After aortic cross-opening,the level of SOD and MMP-2 of the experimental group and the control group were increased (P ＜ 0.05).Before aortic cross-clamping,The activity of SOD in experiment was(85.37 ± 16.82) mU/L,as same as that in the control group((91.51 ± 15.02) mU/L,P ＞ 0.05).The MMP-2 concentration was in experiment group was (362.29 ±29.52) μg/L,as same as that in the control group((372.32 ± 31.42) μg/L,P ＞ 0.05).At 1 minute after aortic cross-opening,there were significant differences regarding of SOD and MMP-2 levels between the both groups(SOD ∶ (106.97 ± 17.46) mU/L vs.(98.74 12.54) mU/L,P ＜ 0.05 ; MMP-2 ∶ (439.48 ± 51.62) μg/L vs.(465.49 ±48.83) μg/L,P ＜0.05) ;The same trend was seen at 15 minutes after aortic cross-opening in two groups in terms of SOD and MMP-2 level (SOD:(104.03 ± 12.63) mU/L vs.(97.94 10.87) mU/L,P ＜0.05; MMP-2:(390.16 ±45.63) μg/L vs.(425.21 ±48.24) μg/L,P ＜0.05).Compare to group B,arrhythmia incidence(x2 =8.223,P ＜ 0.05) and positive inotropic drug dependent degree was lower in group A (x2 =4.022,P ＜ 0.05).Conclusion The cold autologous blood cardioplegia could promote the release of SOD in immature myocardium and reduce the production of MMP-2,which has endogenous protective effect for the infant Imature myocardium.

Objective To study the protective effect of intraaortic protamine injection on lung in infants undergwent opening heart operation by cardiopulmonary bypass surgery. Methods Sixty infants (age ≤ 1 year,weight ≤ 10 kg)who accepted opening heart operation by cardiopulmonary bypass surgery were randomly assigned into 2 groups ( n = 30 in each group) reciving intra-aortic and intra-venous protamine injection respectively. P-peak, P-plate, CL, Oxygenation Index, the number of WBC and neutrophil segregated in lungs were compared between two groups before injecting protamine and 10 minutes, 1 hour, 3 hours after injecting protamine. The time of mechanical ventilation were compared as well. Results P-peak, P-plate, the number of WBC and neutrophil segregated in lungs of intra-aortic injection group significantly decreased than intra-venous injection group at 1 hour, 3 hours after injecting protamine (t =2.743, 3.512; 3.218, 3.469; 3.716, 5.243; 3.853,4. 783 respectively, Ps ＜ 0. 05 ), while the CL and Oxygenation Index increased significantly ( t = 3. 976,4. 267; 4. 557,4. 265 respectively, P ＜ 0. 05 ). The duration of mechanical ventilation follow operation in intraaortic injection group ( [8. 03 ± 5. 14] h ) was shorter compared with intra-venous injection group ( [10. 56 ±6.95]h) (t =2.599,P＜0.05). Conclusion By intra-aortic protamine injection the lung injury decreased significantly. It shows good protective effect on lung in infants underwent opening heart operation by cardiopulmonary bypass surgery.

Objective: To investigate the effect of cold self-blood cardioplegia with ulinastatin on immature myocardial cell apoptosis and protein expressions of Bcl-2, Bax in ventricular septal defect (VSD) infants.
Methods: A total of 60 infants received VSD repairing operation with cardiopulmonary bypass (CPB) in our hospital were summarized. The patients were randomly divided into 2 groups:Test group, the infants received cold self-blood cardioplegia with ulinastatin when aortic cross-clamp was closed. Control group, the infants received cold self-blood cardioplegia when aortic cross-clamp was closed. n=30 in each group. The right atrium tissue was collected before CPB and 10 min after releasing aortic cross-clamp. The index of myocardial cell apoptosis was observed by TUNEL method, and the protein expressions of Bcl-2, Bax were examined by immunohistological method.
Results: Both groups showed the higher index of myocardial cell apoptosis at 10 min after releasing aortic cross-clamp than 5 min before CPB, and the apoptosis index in Test group was lower than that in Control group, all P<0.05. The protein expressions of Bcl-2 and Bax were obviously increased at 10 min after releasing aortic cross-clamp than 5 min before CPB in both groups. Compared with Control group, Test group presented the higher Bcl-2 protein expression and lower Bax protein expression, all P<0.05.
Conclusion: Cold self-blood cardioplegia with ulinastatin could protect immature myocardum from ischemia-reperfusion injury in VSD infants during CPB operation in clinical practice.

Objective To investigate the effects of ulinastatin-containing autologous cold blood cardioplegic solution on the cardiac function of infants after cardiopulmonary bypass surgery.Methods Sixty infants younger than 10 months old,who underwent ventricular septal defect repair under cardiopulmonary bypass,were randomized into autologous cold blood cardioplegia group (30 patients,Group A)and ulinastatincontaining cold blood cardioplegia group (30 patients,Group B).CI,SI and LCWI were monitored 1 and 6 hours after opening the aorta.The time and rate of cardiac resuscitation,as well as the dependence on the inotropic drugs,were intraoperatively monitored.Results The automatic resuscitation rate in two groups was not siynificantly ( P ＞ 0.05).The time for automatic resuscitation were (34.2 ± 4.7) s and (52.1 ± 6.5 ) s for Group B and Group A,respectively ( P ＜ 0.05 ).The rate of dependence on inotropic drug were 40.0％ (12/30) and 66.7％ (20/30)for Group B and Gro~p A,respectively (P ＜ 0.05).Mter the operation,the CI,SI and LCWI of group B were higher than that of group A ( P ＜0.05 ).Conclusion Ulinastatin-containing autologous cold blood cardioplegic solution is beneficial to the functional cardiac recovery of the infants after heart bypass surgery by protecting the immature myocardium.

Objective To observe and evaluate the changes in plasma C3a and C5a concentration after injecting protamine via two different pathways:ascending aorta and superior vena cava.Methods Sixty children with age under 1-year-old who underwent cardiopulmonary bypass were randomly divided into two groups:experimental group(injecting protamine via ascending aorta,n =30)and control group(injecting protamine viasuperior vena cava,n =30;).The plasma concentration of C3a and C5a were measured by ELISA at prior to protamine injected(Time 1)and 1 hour after the protamine injected(Time 2).Results In experimental group,there was no statistical difference on C3a and C5a concentration before and after injection of protamine[C3a:(18.762±3.792) μg/L vs(19.554±3.453) μg/L,t =-0.846,P =0.20; C5a:(0.843±0.159) μg/L vs (0.825±0.119) μg/L,t =0.496,P =0.31].In control group,C3 a concentration increased from(18.780±3.864) μg/L to(22.961±3.501) g/L,C5a concentration increased from(0.839±0.157) μg/L to(0.979±0.116)μg/L after injection of protamine,and the differences were significant(t =-4.392,-3.928,respectively,P ＜0.01).The level of C3a concentration in experimental group was significantly higher than that in control group[(19.554±3.453) μg/L vs.(22.961±3.501) μμg/L,ι =3.795,P < 0.01]after injection of protamine for 1 h and the level of C5a concentration exhibited the same change[(0.825±0.119) μg/L vs.(0.979±0.116)μg/L,t =-5.075,P ＜0.01].Conclusion The levels of C3a and C5a concentration of infants underwent cardiopulmonary bypass are decreased significantly after protamine injected via ascending aorta compated with via superior vena cava.

OBJECTIVETo establish a novel method to determine specific type of amyloidosis through laser microdissection and mass spectrometry (LMD/MS) based proteomic analysis.

METHODSThere were 138 formalin-fixed and paraffin-embedded (FFPE) biopsy samples of patients who were diagnosed as systemic amyloidosis used in this study. For each case, a 10 μm section stained with congo-red and positive amyloid deposits were identified under fluorescent light, followed by micro-dissection and mass spectrometry analysis. The amyloidosis subtype was confirmed based on the most abundant amyloid protein.

RESULTSThe tissue types of 138 specimens were as following: subcutaneous abdominal fat accounted for 26%, tongue for 19%, gingiva for 11%, kidney for 9%, intestine for 9%, heart for 6% and others for 20%. Specific types of amyloid were accurately detected in 121 cases, including 106 (87.6%) amyloid light chain (AL) type, 7 (5.8%) amyloid trans-thy-retin (ATTR), 2 (1.7%) amyloidogenic protein A (AA), 2 (1.7%) amyloid heavy chain (AH)/AL+AH, 2 (1.7%) fibrinogen alpha chain (AFib), 1(0.8%) amyloid apolipoprotein A-type II (AApoA-II) and one (0.8%) amyloid lysozyme (ALys). Diagnosis of amyloidosis was excluded in 5 cases. The types of twelve cases were indeterminate by LMD/MS. On the whole, LMD/MS reached 91.3% accuracy rate in amyloid typing. Commonly involved organs (for example, heart, kidney and liver) turned out to be suitable sources of FFPE samples with typing success rate of almost 100%. In contrast, MS analysis was successful in only 83.3% of subcutaneous abdominal fat samples.

CONCLUSIONLMD/MS method provided a more direct technique for accurate typing of amyloidosis in a single procedure.

Amyloid ; Amyloidosis ; Humans ; Immunoglobulin Light-chain Amyloidosis ; Mass Spectrometry ; Proteomics