Construction of ecological security pattern in Ningbo based on remote sensing ecological index and graph theory knowledge.

Constructing ecological security pattern and identifying ecological important areas are the focus of current research on regional ecological security. With Ningbo City as a case study area, we identified ecological sources by remote sensing ecological index, the ecological corridors and pinch point by circuit theory model, and the minimum spanning tree and cuts by graph theory algorithm. The results showed that there were 203 ecological sources in Ningbo, and that the main type of land cover was forest, including a small amount of paddy fields and flooded vegetation. There were 368 ecological corridors with a total length of 573.42 km, being dense in the southwest and sparse in the northeast. There were 91 ecological pinch points, which mainly distributed between coastal areas and closely related ecological sources. According to current situation, we put forward the optimization strategy with 187 primary corridors, 181 secondary corridors, 50 ecological restoration priority areas and 59 long-term ecological restoration areas. The optimization strategy combined with graph theory and circuit theory model would provide a refe-rence for the constructing of ecological security pattern.

Meta-analysis of risk factors associated with atherosclerosis in patients with Kawasaki disease.

BACKGROUND: Kawasaki disease (KD) has now become the leading cause of acquired heart disease among children in developed countries. This study investigated whether patients with KD have an increased risk of atherosclerosis. METHODS: Electronic databases, including PubMed, Embase and Springer link, were searched through June 1, 2015, for eligible studies. Studies were included when they met the following criteria: 1) an observational study focusing on evaluating the risk factors for atherosclerosis in patients with KD; 2) KD was diagnosed clinically according to the Japan Kawasaki Disease Research Committee or American Heart Association's diagnostic criteria; 3) the study subjects were KD patients without coronary heart disease or related cardiovascular disease (KD group) and non-KD patients as control (control group), and 4) investigation of important atherosclerosis risk factors, total cholesterol (TC), low-density lipoprotein cholesterol (LDL), triglycerides (TG), systolic blood pressure (SBP), and flowmediated dilatation (FMD). The methodological quality of the included studies was evaluated using the Newcastle- Ottawa Scale. Mean difference (MD) and relative risk (RR) and corresponding 95% confidence intervals (CI) were used to calculate the pooled results. RESULTS: Sixteen studies were included with a total of 870 patients, including 421 KD patients and 449 non-KD controls. Differences in TG and SBP between KD patients and controls were not significant; in contrast, TC and LDL levels were significantly higher in KD patients than the controls, whereas FMD in the KD patients was significantly lower. CONCLUSIONS: KD patients may have an increased risk of developing atherosclerosis.

Mibefradil suppresses the proliferation of pulmonary artery smooth muscle cells.

Intracellular Ca(2+) levels play a critical role in the regulation of vasodilation and vasoconstriction by stimulating pulmonary artery smooth muscle cell (PASMC) proliferation, which is important in the pathogenesis of pulmonary arterial hypertension (PAH); however, L-type Ca(2+) channel antagonists are useful in only few patients with PAH. The present study sought to assess the effect of mibefradil, which blocks T-type Ca(2+) channels, on PASMC proliferation and Ca(2+) channel profile. Human PASMCs were stimulated with 25 ng/mL platelet-derived growth factor-BB (PDGF-BB) with and without 10 µM mibefradil or 100 nM sildenafil. After 48 or 72 h, PASMC proliferation and Ca(2+) channel expression were assessed by MTT assays and western blot analysis, respectively. PDGF-BB-induced PASMC proliferation at 72 h (p<0.01), which was inhibited by both sildenafil and mibefradil (p<0.01). Transient receptor potential Ca(2+) channel 6 (TRPC6) expression was significantly increased with PDGF-BB stimulation (p=0.009); however, no changes in TRPC1, TRPC3, CAV1.2, and CAV3.2 levels were observed. Although both TRPC1 and CAV1.2 expression levels were increased in PDGF-stimulated PASMCs on mibefradil and sildenafil treatment, it was not statistically significant (p=0.086 and 1.000, respectively). Mibefradil inhibits PDGF-BB-stimulated PASMC proliferation; however, the mechanism through which it functions remains to be determined. Further studies are required to elucidate the full therapeutic value of mibefradil for PAH.

Coronary artery fistula between single right coronary artery and right pulmonary artery: a case report and literature review.

BACKGROUND: Coronary artery fistula and single coronary artery are two different rare congenital anomalies. The cases with co-existed the two anomalies are more rare. To the best of our knowledge with literature review, the coronary artery fistula between single right coronary artery and right pulmonary artery has not been previously reported. CASE PRESENTATION: In the present article, we report a Chinese patient (a 8-month-old male) who presented cyanosis when cried and heart murmur. The cardiac angiography confirmed coronary artery fistula between single coronary artery arising from the right aortic sinus and right pulmonary artery. Furthermore, the right pulmonary artery was interrupted with main pulmonary artery and the pulmonary blood supplied by single right coronary artery. Following the surgical procedure, the anomalous fistula vessel was cut and sutured. The right pulmonary artery was reconstructed to connect with main pulmonary artery. The patient had an uneventful postoperative course and discharged. Then we reviewed the related literature with Medline and Pubmed databases for further details. CONCLUSION: We believe our patient is the very particular case about coronary artery fistula combined with single coronary artery, and it is first reported with our literature review. As other coronary anomalies, coronary or aortic root angiography is the gold standard method for the diagnosis. Furthermore, early surgery is an optimal treatment in our case.

LMNA gene single nucleotide polymorphisms in dilated cardiomyopathy of Han children.

OBJECTIVE: To investigate whether LMNA gene mutation is associated with dilated cardiomyopathy (DCM) in Chinese Han Race children. METHODS: DNA was isolated from 78 patients with DCM and 100 healthy Chinese children who served as controls. 12 exons in the functional regions and the adjacent part of introns of the LMNA gene were amplified with polymerase chain reactions (PCR) and the PCR products were sequenced with DNA sequencer. We compared the DNA sequence with Blast software online PubMed website. The differences of allele and genotype between the groups were detected by χ(2) test. RESULTS: No disease-causing mutation in LMNA gene was found in all DCM patients. Three nonsense single nucleotide polymorphisms (SNPs) were identified. ① The first is c.1908C>T (H566H, rs4641) which was located at exon 10 of LMNA gene. It was found in 29 DCM cases and 15 control subjects. Compared to healthy controls, the frequency of TT and TC genotypes, and the C allele were significantly increased in DCM patients (P<0.05). ② The second was c.861C>T (A287A, rs5380) which was located at exon 5 of LMNA gene. It was found in 9 DCM cases and 2 control subjects. The frequency of TC genotype was significantly increased in DCM patients (P<0.05). ③ The third was c.1338C>T (D446D, rs5058) which located at exon 7 of LMNA gene. It was found in 8 DCM cases and 3 control subjects. The frequency of TC genotype was significantly increased in DCM patients (P<0.05). CONCLUSION: The SNP of LMNA gene may be associated with the susceptivity of DCM in Chinese Han children.

Evaluating the role of connexin43 in congenital heart disease: Screening for mutations in patients with outflow tract anomalies and the analysis of knock-in mouse models.

BACKGROUND: GJA1 gene encodes a gap junction protein known as connexin 43 (Cx43). Cx43 is abundantly expressed in the ventricular myocardium and in cardiac neural crest cells. Cx43 is proposed to play an important role in human congenital heart disease, as GJA1 knock-out mice die neonatally from outflow tract obstruction. In addition, patients with visceroatrial heterotaxia or hypoplastic left heart syndrome were reported to have point mutations in GJA1 at residues that affect protein kinase phosphorylation and gating of the gap junction channel. However, as these clinical findings were not replicated in subsequent studies, the question remains about the contribution of GJA1 mutations in human congenital heart disease (CHD). MATERIALS AND METHODS: We analyzed the GJA1 coding sequence in 300 patients with CHD from two clinical centers, focusing on outflow tract anomalies. This included 152 with Tetralogy of Fallot from over 200 patients exhibiting outflow tract anomalies, as well as other structural heart defects including atrioventricular septal defects and other valvar anomalies. Our sequencing analysis revealed only two silent nucleotide substitutions in 8 patients. To further assess the possible role of Cx43 in CHD, we also generated two knock-in mouse models with point mutations at serine residues subject to protein kinase C or casein kinase phosphorylation, sites that are known to regulate gating and trafficking of Cx43, respectively. RESULTS: Both heterozygous and homozygous knock-in mice were long term viable and did not exhibit overt CHD. CONCLUSION: The combined clinical and knock-in mouse mutant studies indicate GJA1 mutation is not likely a major contributor to CHD, especially those involving outflow tract anomalies.

[Spatio-temporal expression of connexion (Cx) 40 and Cx45 in Cx43 knockout embryonic mouse hearts].

OBJECTIVE: To investigate the spatio-temporal expression of connexin (Cx) 40 and Cx45 genes in Cx43 knockout embryonic mouse hearts. METHODS: Cx43 knockout heterozygous mice were raised. PCR was performed to identify genotypes of their offsprings. The homozygote (Cx43-/-) was used for study and the wild type (Cx43+/+) was used as control. Immuno-histochemistry was used to detect the Cx40 and Cx45 expression in different parts of the fetal hearts at the embryonic days (EDs) 10.5, 11.5, 12.5, 13.5, 14.5, and 15.5, respectively. SCIM microscopic image analytic system was used for quantitative analysis of staining intensity. RESULTS: (1) Cx40 expression was detected in ventricles of Cx43+/+ fetal heart as early as ED10.5 with the intensity represented by A value of 8.6. Subsequently it was distributed in the atria and ventricles with the peak expression observed at ED14.5 (A value = 94.8), and faded afterwards. Less Cx40 expression was observed in the Cx43-/- fetal hearts as compared with Cx43+/+ although its expression pattern was similar in both groups. (2) Cx45 expression was detected in ventricles at ED 10.5 (A value = 20.0). It was subsequently distributed in the atria and ventricles with the peak expression observed at ED12.5 (A value = 49.6), and then faded. Less Cx45 expression was observed in the Cx43-/- fetal hearts as compared with Cx43+/+ although its expression pattern was similar as well in both groups. CONCLUSION: Down-regulated expression of the genes Cx40 and Cx45 may be associated with the abnormal heart development in Cx43 knockout animals.

[Abnormal development of conotruncal region in Cx43 knockout mice].

OBJECTIVE: To explore the etiology of the conotruncal malformations in Cx43 knockout mice. METHODS: The objects were C57/BL6 mice of E11.5 to 1 day after birth by the mating of 2 month old heterozygous mice which included Cx43 (knockout, KO) homozygotes (Cx43-/-), heterozygotes (Cx43+/-) and wild-types (Cx43+/+) genotyped by PCR method. Microdissection and HE staining were used to examine the structures of hearts. The expression of the alpha-SCA, alpha-SMA, AP-2alpha were detected by immunohistochemistry. AP-2alpha mRNA was detected by in situ hybridization. RESULTS: Cx43-/- mice died within 24 h after birth with a swelling and blockage of the conotruncal region, which led to the obstruction of OFT and enlargement of right ventricle. HE staining showed plenty of abnormal tissues in this region forming many pouches. No apparent malformations were observed in Cx43+/- and Cx43+/+ mice. The expression of alpha-SCA in the proximal OFT septum was delayed obviously in Cx43-/- predominantly at E13.5 and E14.5. The expression of alpha-SMA in the OFT in Cx43+/- and Cx43-/- was stronger than that of Cx43+/+ mice, and mostly located in the hyperplastic conotruncal region especially at E13.5-E15.5 in Cx43-/- mice. The expression could still be observed at the birth day in Cx43-/- mice, which was not observed in Cx43+/+ mice. The expression of AP-2alpha and AP-2alpha mRNA at E13.5 increased in Cx43-/- and abnormally located in the proximal OFT septum. CONCLUSION: Cx43 KO mice are characterized by hyperplasia in conotruncal region. Cx43 KO mice exhibited a delayed myocardialization and the developmental immaturity of cardiomyocytes. The abnormal distribution of cardiac neural crest cells is likely to contribute to the conotruncal malformations in Cx43-deficient mice.

[Histopathological analysis of neonatal mouse hearts with connexin43 gene defects].

OBJECTIVE: To investigate the characteristics of heart morphology in neonatal mice with connexin43 gene defects. METHODS: Two mouse lines were used in this study, which included connexin43 knockout (Cx43KO) mice and CMV43(CT) transgenic mice. PCR analysis was carried out to identify the genotypes of two transgenic mice. Sections with HE staining were analyzed to display the morphologic structures of the hearts in neonatal mice. C57BL6/SJ mice were used as control. RESULTS: All 11 homozygous Cx43KO mice died within 24 hr after birth showing severe right ventricular outflow tract obstruction (RVOTO). Out of 20 homozygous CMV43(CT) transgenic mice, 12 mice died within 48 hr after birth showing not only RVOTO, but also atrial septal defect (ASD), ventricular septal defect (VSD) and other cardiac defects, while the remaining 8 mice were alive without heart defects. Both heterozygous Cx43KO and CMV43(CT) transgenic mice had normal hearts. CONCLUSION: Connexin43 gene defect is obviously associated with abnormal heart morphogenesis. The different types and different dosage of the gene defects may lead to different phenotypes of hearts.

Mutations of connexin43 in fetuses with congenital heart malformations.

BACKGROUND: Gap junction channels formed by connexin43 (Cx43) protein are important in cardiac morphogenesis, and Cx43 gene is thought to be associated with congenital heart malformation (CHM). This study was undertaken to detect the mutations of Cx43 in fetuses with CHM. METHODS: Cx43 extron DNA was amplified by PCR from 16 fetuses with a variety of CHM. The PCR products were analyzed by SSCP and DNA sequencing. Thirty children who had no CHM were selected as controls. RESULTS: Eight homozygous mutations of Cx43 were observed in a fetus with double outlet right ventricule (DORV), five of the 8 mutations were missense mutations including Arg239Trp, Ser251Thr, Ala253Pro, Pro283Leu and Thr290Asn, and the remaining 3 were silent polymorphisms including Gly252Gly, Pro256Pro and Thr275Thr. No mutations were found in other fetuses and the control group. CONCLUSIONS: Mutations of Cx43 may be associated with congenital conotruncal anomalies. PCR-SSCP is an effective method for screening the mutations of Cx43.