Andrographolide inhibits NF-kappaBeta activation and attenuates neointimal hyperplasia in arterial restenosis.

The NF-kappaBeta transcription factors modulate the expression of tissue factor (TF), E-selectin (CD62E) and vascular cell adhesion molecule-1 (VCAM-1), which are essential for thrombosis and inflammation. We have previously shown that andrographolide (Andro) covalently modifies the reduced cysteine(62) of p50 - a major subunit of NF-kappaBeta transcription factors, thus blocking the binding of NF-kappaBeta transcription factors to the promoters of their target genes, preventing NF-kappaBeta activation and inhibiting inflammation in vitro and in vivo. Here we report that Andro, but not its inactive structural analog 4H-Andro, significantly suppressed the proliferation of arterial neointima ( approximately 60% reduction) in a murine model of arterial restenosis. Consistently, p50(-/-) mice manifested attenuated neointimal hyperplasia upon arterial ligation. Notably, the same dosage of Andro did not further reduce neointimal formation in p50(-/-) mice, which implicates the specificity of Andro on p50 for treating experimental arterial restenosis. The upregulation of NF-kappaBeta target genes, including TF, E-selectin and VCAM-1, and the increased deposition of leukocytes (mainly CD68+ macrophages) were clearly detected within the injured arterial walls, all of which were significantly abolished by treatment with Andro or genetic deletion of p50. The expression of TF, E-selectin and VCAM-1 was also markedly upregulated in the patient sample of thrombotic vasculitis, indicating the clinical relevance of NF-kappaBeta activation in the pathogeneses of occlusive arterial diseases. Our data thus indicate that, by the downregulation of the NF-kappaBeta target genes that are critical in thrombosis and inflammation, specific inhibitors of p50, such as Andro, may be therapeutically valuable for preventing and treating thrombotic arterial diseases, including neointimal hyperplasia in arterial restenosis.

Surgical repair of sinus of Valsalva aneurysm in Asian patients.

BACKGROUND: Sinus of Valsalva aneurysm is a rare cardiac anomaly, and the difference between Asian and Western countries in its occurrence is not well established. This study was designed to investigate the difference between Asian and Western patients. METHODS: Between September 1988 and February 2006, 83 patients with sinus of Valsalva aneurysm underwent surgical repair in our institute. The aneurysms originated from the right and noncoronary sinus in 74 and 9, respectively, and ruptured into the right ventricle in 52 patients, the right atrium in 30, and the left ventricle in 1. Ventricular septal defect (n = 38), aortic regurgitation (n = 21), and bicuspid aortic valve (n = 4) were the common coexisting anomalies. To compare the differences between Asian and Western patients in sinus of Valsalva aneurysm, 1049 cases (654 Asian patients versus 395 Western) were collected from the literature. RESULTS: Sixty-six patients were followed up for 9.6 +/- 3.8 years. The cardiac function of 15 patients with aortic regurgitation was worse than that of those with no aortic regurgitation (p < 0.05). There was no difference between the direct closure and the patch closure (p > 0.05). Analysis of all collected cases revealed that aneurysm of the sinus of Valsalva in Asian patients compared with Western series is characterized by a higher incidence, more aneurysms originating from the right coronary sinus (85.8% versus 67.9%), more aneurysm rupture into the right ventricle (72.5% versus 60%), a higher incidence of association with ventricular septal defect (52.4% versus 37.5%), and lower incidence of association with bicuspid aortic valve (0.6% versus 7.8%). However, both Asian and Western patient series have similar incidence of combination with aortic regurgitation (33.6% versus 32.7%). CONCLUSIONS: Long-term results of ruptured sinus of Valsalva aneurysm are associated with preoperative aortic regurgitation. The difference between Asian and Western patients with ruptured aneurysm of the sinus of Valsalva is significant.

[The effects of coronary abnormalities on reconstruction of right ventricular outflow tract in patients with complex congenital heart disease].

OBJECTIVE: To explore the effects of coronary abnormalities on reconstruction of right ventricular outflow tract in patients with complex congenital heart disease. METHODS: Eighty patients, including tetralogy of Fallot 14 cases and double outlet right ventricle (DORV) 4 cases with anomalous coronary artery, underwent surgical treatment from April 1989 to May 2004. The coronary diseases included single left or right coronary artery in 11 cases, left anterior descending coronary artery originating from right coronary artery in 6 cases, vascular plexus on right outflow tract in 1 case. One-stage correction was undergone in 17 cases, palliative procedure in 1 case. The reconstruction methods were as follows: mobilizing coronary artery and expanding incision under coronary artery with pericardial patches in 4 cases; repairing ventricular septal defect (VSD) via the incision beneath the coronary artery and expanding right ventricular outlet tract (RVOT) and pulmonary via longitudinal incision over the coronary artery in 3 cases. VSD were repaired via right atrium and minimal incision on RVOT plus incision on pulmonary were made to expand RVOT. Trunk of pulmonary were anastomosis with RVOT in 3 cases. Homograft valved aorta were used in 3 cases. RESULTS: One case died of serious low cardiac output syndrome postoperatively. There was no critical complication of hemorrhage and respiratory tract. Oxygen saturation rose from 68.0% to 82.0% after treated by palliative procedure. Seventeen cases were followed from 10 months to 8 years, 1 case suddenly died with no clear cause. Three cases were with residual leak, 3 with residual obstruction. CONCLUSIONS: Surgical procedure should be selected according to the characteristics of coronary disease with complex congenital heart disease.

[Construction of vector tandem expressing rat ventricular myocyte Kir2.1 shRNA and its effect in vitro].

OBJECTIVE: To construct a recombinant vector that expresses 5 shRNAs targeting on the rat ventricular myocyte Kir2.1 gene in tandem and its effect in vitro METHODS: Ventricular myocytes were collected from newborn Wistar rats and cultured. Five sites targeting on the rat Kir2.1 gene were selected. Accordingly 5 pairs of oligonucleotide fragments were designed, synthesized, and annealed to obtain double-stranded DNAs. The 5 pairs of oligonucleotide were then cloned into the vector pGenesil-1 by repeated excision and ligation successively. The tandem recombinant vector pEGFP6-1Kir2.1 was thus constructed and transfected into the cultured rat myocytes. RT-PCR and Western blotting were used to detect the mRNA and protein expression of Kir2.1 in the myocytes. Sequence not related to Kir2.1 sequence with mismatched bases was designed and used as control. RESULTS: A recombinant vector that expresses 5 shRNAs targeting on the rat ventricular myocyte Kir2.1 gene in tandem was constructed. 96 hours after the transfection RT-PCR showed that the Kir2.1 mRNA transcription was suppressed by 83, 6%, and Western blotting showed that the Kir2.1 protein transcription was suppressed by 68.1% in comparison with the control. CONCLUSION: The vector that expresses the 5 shRNAs targeting on the rat ventricular myocyte Kir2.1 gene in tandem is able to suppress the expression of Kir2.1 in rat ventricular myocytes. Application of such vector may be a new method to produce a new type of heart biological pacemaker.

[Clinical analysis for 70 sinus of valsalva aneurysm].

OBJECTIVE: To review retrospectively the experience of surgical repair of sinus of valsalva aneurysm (SVA) in 70 patients. METHODS: Between September, 1988 and October, 2003, Seventy patients with SVA underwent surgical repair by the aid of general anesthesia and cardiopulmonary bypass, comprised 1.4% (70/4960) of all open-heart operation. Forty-five were male and 25 female. Age ranged from 3 to 69 years old [mean (29 +/- 15) years]. The aneurysms ruptured into the right ventricle in 46 patients, right atrium in 23 and left ventricle in 1 respectively. The aneurysms originated from right and noncoronary sinus in 61 patients (87%) and 9 patients (13%) respectively. The most common associated cardiovascular lesions were ventricular septal defect (VSD, n = 34) and aortic valve incompetence (n = 21). Repairs were achieved through an incision in right atriotomy, right ventriculotomy or aortotomy only or both aortotomy and right atriotomy (or right ventriculotomy). The defects in the sinus of valsalva was repaired with either direct sutures (n = 43) or a patch (n = 27). The aortic valve was replaced in 6 patients. RESULTS: There were no deaths in early time after repair. Postoperative hospital stay was 8 approximately 33 days [mean +/- standard deviation, (14.3 +/- 6.4) days] before 1997 and 6 approximately 15 days [mean +/- standard deviation, (9.1 +/- 2.6) days] after 1997 respectively. Complications included infection (n = 4), hemorrhage (n = 4), pneumothorax (n = 1), arrhythmia (n = 4) and residual shunt (n = 1) of VSD. Fifty-three (76%) patients (2 months approximately 13 years) were followed-up [mean +/- standard deviation, (6.6 +/- 3.8) years]. All patients survived except that one died of rupture of dissecting aortic aneurysm 7 years after operation. CONCLUSIONS: The ruptured sinus of valsalva aneurysm and unruptured sinus of valsalva aneurysm with ventricle septal defect or(and) aortic valve regurgitation should be repaired surgically as soon as the diagnosis was confirmed. Long-term results are associated with preoperative aortic valve regurgitation.