[Effects of immunosuppressive treatment in prevention of calcification in aortic valved homograft: experiment with rats].

OBJECTIVE: To investigate the effects of immunosuppressive treatment in prevention of calcification in aortic valved homograft (AVH). METHODS: 120 Wistar rats were randomly divided into 4 equal groups: Group A (allogene group) undergoing incision of the abdominal aorta and implantation of the AVH with myocardial cuff from SD rats, Group B, injected with cyclosporine A intraperitoneally one day after the implantation, Group C, injected intraperitoneally with anti-dendritic cell monoclonal antibody (DCmAb) one day after the implantation, and Group D (isogenenic or control group), receiving the AVH of another Wistar rats. All groups were further subdivided into 5 equal subgroups to be sacrificed at different time points: 2, 4, 8, 12, and 16 weeks postoperatively. Blood samples were obtained from the vena cava to detect the expression of T-cell antigen receptor (TCR)-alpha and beta and CD28 by flow cytometry. AVH specimens were obtained to observe the changes of endotheliocytes and smooth muscle cells with light and electron microscopy. The expression of CD54 was detected by immunohistochemistry. The calcium content of the AVH tissue after transplantation was examined by flame atomic absorption spectrophotometry. RESULTS: (1) Compared with the isogenic group, the expression levels of TCR-alpha and beta and CD28 in the allogener groups were all significantly higher at all time points (all P < 0.01), peaked 2 approximately 4 weeks after operation, then gradually decreased, and approached the level of the controls 12 weeks after the implantation. Specifically, the expression levels of TCR-alpha and TCR-beta 2 and 4 weeks postoperatively of Group A were 52.4% +/- 3.3% and 43.8% +/- 6.4% respectively, significantly higher than those of Group B [(34.5 +/- 3.5)% and (31.6 +/- 2.6)% respectively], Group C [(31.6 +/- 2.3)% and (29.5 +/- 3.0)% respectively), and Group D (23.2 +/- 1.3)% and (21.6 +/- 2.3)% (all P < 0.01)]; and the CD28 expression level 2 approximately 4 weeks after operation of Group A were (51.7 +/- 7.5)% and (66.3 +/- 4.4)% respectively, both significantly higher than those of Group B [(41.2 +/- 1.6)% and (55.1 +/- 5.1)% respectively], Group C [(36.6 +/- 3.6)% and (51.8 +/- 5.6)% respectively], and Group D [30.7 +/- 1.4)% and (33.3 +/- 0.9)% respectively)] [all P < 0.01 except those levels 12 and 16 weeks after the operation in each subgroup (P > 0.05)] And the levels of TCR-alpha and TCR-beta and CD28 of the 2 treatment groups were all significantly lower than those of the untreated group (Group A) (all P < 0.01). (2) The calcium contents of the AVH tissues of Group A, B, and C significantly increased 4 weeks after the operation and peaked 12 and 16 weeks after operation. No significant difference in calcium level was found in Group D at different time points (all P > 0.05). The calcium contents in AVH tissues 4 and 8 weeks postoperatively of Groups A, B, and C were (2856 +/- 79) microg/g and (3587 +/- 168) microg/g respectively, (2518 +/- 73) microg/g, (3237 +/- 187) microg/g; and (2176 +/- 210) microg/g and (3089 +/- 176) microg/g; all significantly higher than those of Group D (860 +/- 60) microg/g and (870 +/- 50) microg/g respectively, all P < 0.01. CONCLUSION: Immunosuppressive treatment obviously reduces the immune rejection and delays the course of AVH calcification.

[Clinical values of detection of serum levels of S100B and NSE in diagnosis of brain injuries at early period after cardiopulmonary bypass].

OBJECTIVE: To investigate the clinical values of detecting the blood serum levels of S100B and neuron-specific enolase (NSE) in diagnosis of brain injuries at early period after cardiopulmonary bypass (CPB). METHODS: Forty-eight patients with heart disease were divided into 2 groups: CPB group (n = 40) and off-CPB group (n = 8). Before operation, and 24 hours and 48 hours after CPB specimens of peripheral blood were collected and ELISA was used to detect the serum levels of S100B and NSE. Forty-eight hours after the operation brain damage quotient (DQ) was calculated. RESULTS: The serum levels of S100B 24 and 48 hours after operation of the CPB group were 0.61 microg/L +/- 0.18 microg/L and 0.37 microg/L +/- 0.12 microg/L respectively, both significantly higher than that before the operation (0.05 microg/L +/- 0.03 microg/L, P < 0.001). The serum levels of NSE 24 and 48 hours after operation of the CPB group were 10.14 microg/L +/- 3.87 microg/L and 5.77microg/L +/- 2.31 microg/L respectively, both significantly higher than that before operation (2.98 microg/L +/- 1.49 microg/L, P < 0.001). The serum levels of S100B 24 and 48 hours after operation of the off-CPB group were 0.05 microg/L +/- 0.03 microg/L and 0.04 microg/L +/- 0.03 microg/L respectively, both not significantly different from that before operation (0.04 microg/L +/- 0.03 microg/L, P > 0.05). The serum levels of NSE 24 and 48 hours after operation of the off-CPB group were 2.91 microg/L +/- 1.56 microg/L and 2.87 microg/L +/- 1.41 microg/L respectively, both not significantly different from that before operation (2.76 microg/L +/- 1.23 microg/L, P > 0.05). The levels of S100B and NSE 24 hours after CPB were positively correlated with age, CPB time, and cross-clamp time (all P < 0.05). The levels of S100B and NSE 48 hours after CPB were positively correlated with the brain DQ (r = 0.739 P < 0.01, r = 0.371 P < 0.05). The multiple correlation coefficient square (R2) of detection of the levels of both S100B and NSE was 0.851, significantly higher than that of mere detection of S100B (R2 = 0.703) and that of mere detection of NSE (R2 = 0.482) (both P < 0.01). CONCLUSION: Both serum S100B and serum NSE are sensitive markers in the early diagnosis of brain injuries after CPB. Detection of both S100B and NSE is the most specific, and mere detection of S100B comes behind.

[Effects of blood washing and autotransfusion during cardiopulmonary bypass on erythrocyte immune and kidney function].

OBJECTIVE: To study changes of erythrocyte immune and kidney function after autotransfusion washed red blood cells during cardiopulmonary bypass (CPB). METHODS: Thirty-two patients undergoing valve replacement with CPB were randomly divided into study group and control group (16 in each group). In study group, the blood in operative field and the residual blood in the extracorporal machine were collected, centrifuged, washed and retransfused to patients. Patients in control group were transfused with the residual blood in the extracorporal machine without any disposal or banked blood. All patients were used with membrane oxygenator. Before CPB, 12 h, 24 h, 72 h and 7 d after CPB, whole blood were taken, then the erythrocyte immune function (C3bRR, RICR) and level of plasma free hemoglobin (FHB) were assayed, and post-operation renal function was compared between the two groups. Moreover, total volume of banked blood transfused to patients after CPB was recorded. RESULTS: (1) After 12 hours, 24 hours, 72 hours, 7 days of CPB, the RBC-C3bRR (14.3% +/- 4.7%, 15.9% +/- 3.6%, 16.6% +/- 2.8%, 19.9% +/- 4.1%) and RBC-ICR (8.7% +/- 1.9%, 9.2% +/- 2.0%, 9.5% +/- 2.6%, 12.0% +/- 2.0%) in study group were significantly elevated than that in control group (RBC-C3bRR 10.7% +/- 2.4%, 11.3% +/- 3.0%, 12.3% +/- 3.5%, 14.5% +/- 2.0%, RBC-ICR 5.9% +/- 1.4%, 6.0% +/- 1.8%, 7.0% +/- 1.7%, 8.7% +/- 2.7%). The erythrocyte immune function after CPB was better and restored faster in study group than that in control group (P < 0.05 in all). (2) After 12 hours, 24 hours of CPB, the levels of FHB (0.41 g/L +/- 0.13 g/L, 0.03 g/L +/- 0.02 g/L) in study group were significantly lower than that in control group (1.02 g/L +/- 0.23 g/L, 0.54 g/L +/- 0.09 g/L) (P < 0.01). After 24 hours of CPB, the level of urinary protein excretion (0.19 g/d +/- 0.08 g/d) in study group was significantly lower than that in control group (0.32 g/d +/- 0.07 g/d) (P < 0.05). (3) After 24 hours of CPB, the level of 24 h creatinine clearance was significantly elevated in study group (68 ml x min(-1) x 1.73 m(-2) +/- 10 ml x min(-1) x 1.73 m(-2)) than that in control group (45 ml x min(-1) x 1.73 m(-2) +/- 4 ml x min(-1) x 1.73 m(-2)) (P < 0.01). (4) The total volume of banked RBCs transfused after CPB were fewer in study group (2.0 U +/- 1.1 U) than that in control group (7.4 U +/- 2.3 U) (P < 0.01). CONCLUSION: Autotransfusion of washed red blood cells during CPB may improve significantly the erythrocyte immune function and protect kidney function better than transfusion of residual blood in the extracorporal machine or banked blood.

[Analysis of the factors which influence allograft calcification after aorta transplantation of allogenetic rat].

AIM: To explore the factors which influence the calcification of homograft after aorta transplantation of allogenetic rat. METHODS: The research was devided into 2 groups: allogene group and isogenenic group. Allogene group: SD -->Wistar. Isogenenic group: Wistar to Wistar. Aortic valve homograft was heterotopically allografted onto abdominal aorta. The rats were sacrificed in batches at 2, 4, 8, 12 and 16 weeks postoperatively. Blood samples were obtained for accessing the expression of CD25, CD71, and AVH was obtained for accessing the calcium level and the expression of CD40. At the same time, the change of endotheliocyte and smooth muscle cells were observed with transmission electron microscope. RESULTS: (1)Compared with the isogene group, the expression of CD40, CD25 and CD71 in allogene group was much higher at each time point and reached peak at 2-4 weeks after operation. (2)The calcium level in allogene group increased at 4 weeks after operation and reached the peak at 12 weeks after operation. No significant difference in calcium level was found in isogenenic group over 5 different periods. (3)Exfoliation of endotheliacytes as well as necrosis of smooth muscle cells were observed in the graft in allogene group. CONCLUSION: The calcium level of homograft had a relation with immunological rejection. The calcification began at 4 weeks postoperation; the calcium level of homogaft increased gradually and reached the peak at 12 weeks postoperation and then maintained on a stable level.

[Study on the effect of adrenomedulin and urotensin-II on pulmonary hypertension of patients with congenital heart disease].

OBJECTIVE: To evaluate the effect and clinical significance of adrenomedulin (ADM) and urotensin-II (UII) on pulmonary hypertension (PH), by detecting their levels of patients with congenital heart disease and pulmonary hypertension. METHODS: 52 patients with congenital heart disease who had left-to-right shunt were selected randomly. 52 patients were divided three groups according to pulmonary artery systolic pressure (PASP): group A was not pulmonary hypertension (PASP < 30 mm Hg, n = 17); group B was mild pulmonary hypertension (PASP30-49 mm Hg, n = 18); group C was moderate and severe pulmonary hypertension (PASP > or = 50 mm Hg, n = 17). The plasma levels of ADM and UII were detected at each period by radioimmunoassay (RIA) method. It was analyzed the changes of their levels within pre-operation, 20 mins and 7 days post-operation and the interrelation between them and PASP. RESULTS: (1) Following the severity degree of pulmonary hypertension, the plasma levels of ADM increase. There is positive correlation between PAP and plasma level of ADM (pre-operation r = 0.8012, P < 0.01; 20 min post-operation r = 0.6325, P < 0.01; 7 d post-operation r = 0.7126, P < 0.01). (2) Following the severity degree of pulmonary hypertension, the plasma levels of UII don't change obviously. There is no correlation between PAP and plasma level of UII (P > 0.05). (3) The plasma levels of ADM: group A (pre-operation: 33 pg/ml +/- 5 pg/ml, 20 mins post-operation: 29 pg/ml +/- 4 pg/ml, 7 d post-operation: 20 pg/ml +/- 3 pg/ml); group B (pre-operation: 44 pg/ml +/- 8 pg/ml, 20 mins post-operation: 40 pg/ml +/- 6 pg/ml, 7 d post-operation: 34 pg/ml +/- 4 pg/ml); group C (pre-operation: 60 pg/ml +/- 10 pg/ml, 20 mins post-operation: 58 pg/ml +/- 8 pg/ml, 7d post-operation: 38 pg/ml +/- 4 pg/ml). Plasma level of ADM of each group after CPB is lower than that of each group before operation. It is statistical difference only 7 days post-operation (group A q = 5.41, P < 0.01; group B q = 4.76, P < 0.01; group C q = 6.32, P < 0.01). (4) The plasma levels of UII: group A (pre-operation: 2.2 pmol/L +/- 0.5 pmol/L, 20 mins post-operation: 2.2 pmol/L +/- 0.44 pmol/L, 7 d post-operation: 2.2 pmol/L +/- 0.6 pmol/L); group B (pre-operation: 2.7 pmol/L +/- 0.6 pmol/L, 20 mins post-operation: 2.6 pmol/L +/- 0.6 pmol/L, 7 d post-operation: 2.6 pmol/L +/- 0.5 pmol/L); group C (pre-operation: 2.9 pmol/L +/- 0.6 pmol/L, 20 mins post-operation: 2.6 pmol/L +/- 0.7 pmol/L, 7 d post-operation: 2.8 pmol/L +/- 0.4 pmol/L). Compared with that of each group before operation, Plasma level of UII of each group after operation is no obvious difference (P > 0.05). CONCLUSION: (1) Following the severity degree of pulmonary hypertension, the plasma levels of ADM increase. ADM plays an important role in the formation of pulmonary hypertension and restructure. (2) Following the severity degree of pulmonary hypertension, the plasma levels of UII don't change obviously. There is no correlation between PAP and plasma level of UII, but UII may be play an important role in the formation of pulmonary hypertension and restructure. (3) Measuring the levels of ADM may be a reliable method to follow the change of pulmonary pressure and worsening of pulmonary hypertension.

[Application of one-way valved patch in treatment of patients with congenital heart disease with severe pulmonary hypertension].

OBJECTIVE: To explore the effect of one-way valved patch used in congenital heart disease with severe pulmonary hypertension. METHODS: One-way valved patch was used in 30 patients of congenital heart disease with severe pulmonary hypertension (PP/PS > 0.75) in operation. Follow-up of 6 approximately 86 months was conducted to observe its effect. RESULTS: The pulmonary artery pressure was significantly decreased without trans-patch shunt in 11 cases postoperatively. Trans-patch shunt was determined in 27 cases within postperative 72 hours. There were 2 postoperative deaths out of these 27 patients: one died of low cardiac output syndrome 72 hours after operation, and the other died of right heart failure 4 weeks after operation. Thirty-six patients were restored to health and discharged. Three-month follow-up showed trans-patch shunt in 7 cases, including right-to left shunt in 4 cases and two-side shunt in other 3 cases. Color Doppler ultrasonography conducted 6 months after operation proved trans-patch shunt in 4 cases, right-to-left shunt in 1 case, two-side shunt in 2 cases, and left-to-right shunt in 1 case (PP/PS = 0.45). CONCLUSION: One-way valved patch is useful in selected patients in which postoperative right heart failure can be anticipated so as to shunt the blood in the right heart to the left heart and increase the blood volume in the left heart system to ensure the left heart output and minimize the risk of postoperative right heart failure, at the expense of systemic low oxygen saturation that is, however, well tolerated.