Neuro death through autophagy via the acetylation of FoxO1 by SIRT2 in the hippocampus of mice in a autism spectrum disorder mice model.

Autism Spectrum Disorder (ASD) is a series of complex neurodevelopmental disorders, which can affect children's social, behavioral and communication abilities. A member of the Sirtuins family of NAD + dependent deacetylases called SIRT2 could regulate the inflammation progress during stress, but the relevant mechanism has not been clearly defined. In the present study, the ASD model of wild type and SIRT2 knock out mice was established to evaluate the impact on the homeostasis of neurons in the hippocampus using western blotting, immunofluorescence and Nissl staining. The results showed that the amplification of neuronal richness was significantly decreased and neuroinflammation increased in the hippocampus following ASD due to autophagy, caused by enhancing the acetylation of FoxO1 using SIRT2 gene deletion and indicating this should be the target for ASD or other psychological stress treatment.

Repeated social defeat stress inhibits development of hippocampus neurons through mitophagy and autophagy.

The social defeat stress model is commonly used to study depression and anxiety disorder, which can significantly affect the structure and function of neurons in the hippocampus; however, the relevant mechanism in neuronal loss has not been clearly defined. In the present study, a social defeat stress model was established in mice to evaluate the impact of social defeat stress on the structure of neurons in the hippocampus using Western blotting, immunofluorescence, Nissl staining, Golgi staining and transmission electron microscopy. The results demonstrated that social defeat stress leads to disruption of homeostasis in the hippocampus and the integrity of mitochondria in hippocampal neurons was markedly affected by enhanced mitophagy and autophagy resulting in inhibition of development and growth. These findings provide new insights into the mechanisms of neuronal development and growth due to social defeat stress, which should help in the development of new strategies to combat the effects of depression and anxiety disorder.

[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 of 24 supracardiac total anomalous pulmonary venous connection treated with surgery].

OBJECTIVE: To review the surgical treatment of supracardiac type of total anomalous pulmonary venous connection (TAPVC). METHODS: Twenty-four patients with supracardiac total anomalous pulmonary venous connection underwent surgical correction from July 1995 to June 2004. There were 11 males, 13 females. The patients aged from 40 days to 35 years (mean 4.5 years). Twenty-three patients were under 6 years old. The weight was from 3.8 to 54.0 kg (mean 17.5 kg). Ten patients were through right atrial incision, 8 through double-atrium incision to anastomose the left atria and the common pulmonary veins. Two used large patches separating the portal of the common pulmonary veins and the enlarged atrial septal defects to the left atria. The left atrium and the common pulmonary veins were anastomosed in one case using a large patch separating the portal of the common pulmonary vein and the enlarged atrial septal defect to the left atrium. Three anastomosed the top of the left atria and the common pulmonary veins. RESULTS: One patient died postoperatively of low cardiac output syndrome. One with cardiac tamponade after operation was cured by reoperation. Six with arrhythia were all cured. One with acute pulmonary edema was cured by reopen of the vertical vein and closed later. The postoperative follow-up period ranged from 3 months to 7 years (mean 2.5 years). One complicated severe supracardiac arrhythia 2 weeks later. One with late pulmonary occlusion was cured by reoperation. There was no long term complication of the others. CONCLUSIONS: The operative methods of TAPVC depend on the cardiac deformation. Proper surgery timely treatment of the postoperative complications may achieve a satisfactory result. The method of only use patch in the right atrium should select proper repair material. Anastomosing the top of the left atrium and the common pulmonary vein is a good method. It is easy for exposure and operating, meanwhile it has low occurrence of arrhythmias and should be recommended.

Adenovirus mediated endothelial nitric oxide synthase gene transfer prevents restenosis of vein grafts.

The possibility of endothelial nitric oxide synthase (eNOS) gene transfer, which prevents restenosis of vein grafts, was explored in 16 goats. The recombinant adenoviral vector coding endothelial nitric oxide synthase (AdCMVeNOS) and adenoviral vector (AdCMV) were constructed. A total of 6 cm jugular vein was removed, cut into two equal lengths for vein grafts, and infected with AdCMVeNOS or AdCMV in vitro. One segment (2 cm) of each carotid artery was removed. The vein graft that had been infected with AdCMVeNOS was anastomosed to the right carotid artery, and the vein graft that had been infected with AdCMV was anastomosed to the left. The functional expression of eNOS in vein grafts was assessed by the immunohistochemical staining and measurement of NO concentration. The inhibition of intimal hyperplasia in vein grafts was evaluated by the assay of 3H-TDR incorporation, histologic analysis, measurement of intimal thickness, and percent area stenosis. Adenovirus mediated eNOS gene transfer to goat vein grafts resulted in functional transgene expression with increased NO release. Increased local NO production could inhibit intimal hyperplasia and increase the patent rate of vein grafts.