ABSTRACT: Rhododendron fortune belongs to a scented Rhododendron species native to China, which produces fragrant flowers of great ornamental and environmental values for landscaping or indoor beautification. However, the scents in Rhododendron fortuneihave not yet been investigated, let alone the mechanism of the formation of these fragrance in the flowers. In this study, we measured the scents in terms of its volatile components and contents (VOC) in Rhododendron fortuneiat four different flowering stages and in different tissues of the plant by headspace solid-phase micro-extraction combined (HS-SPME) with gas chromatography-mass spectrometry (GC-MS). Then the characteristic aromatic values, which reflects the degree of scent perception by human, of each VOC in the plant was calculated according to its respective aromatic thresholds. Results showed that three main VOCs measured from highest to lowest are methyl benzoates, terpenes and fatty acid derivatives. Their content increased after the flower bud opening and reached the highest at half to full blossom. In a flower most VOC contents were measured in petals and only trace amount in other tissues such as stamen, pistil. A small amount of VOCs was determined in leaves as well.All aromatic values were almost corresponded to the contents of three main VOCs, indicating that the flower fragrance arises truly from these VOC components. S-adenosyl-L-methionine: benzoic acid carboxyl methyl transferase (BAMT) catalyzes the final step to form methyl benzoates. To understand the mechanism of the formation of this main type fragrance and its regulation, we firstly isolate a gene of RfBAMT from petal of Rhododendron fortuneiby using homologous cloning and RACE technology. The full length of its cDNA was 1383 bp，with an open reading frame of 1104 bp, encoding a total of 368 amino acids. The phylogenetic tree analysis showed that RfBAMT was the closest to the BSMT of Camellia japonica, belonging to methyltransferases family. Then we measured the expression level of RfBAMT again at four flower developmental stages and in different flower tissues and leaves. The results showed that the expression level of this gene was highly positively correlated with the emitted content of methyl benzoates in the flowering, implying that RfBAMT plays a pivotal role in the formation and regulation of methyl benzoates in Rhododendron fortune.Thisresearchshowed that the RfBAMT was cloned and identified in our study and its expression level was highly positively correlated with the emitted content of methyl benzoates in the flowers and leaves, which indicated this gene may play an important role on regulation of methyl benzoate synthesis in Rhododendron fortunei.

RESUMO: Rhododendron fortunei pertence a uma espécie de rododendro perfumada nativa da China, que produz flores perfumadas de grande valor ornamental e ambiental para paisagismo ou embelezamento de interiores. No entanto, os aromas em Rhododendron fortunei ainda não foram investigados, muito menos o mecanismo de formação dessas fragrâncias nas flores. Neste estudo, medimos os aromas em termos de seus componentes e conteúdos voláteis (VOC) em Rhododendron fortunei em quatro diferentes estágios de floração e em diferentes tecidos da planta por microextração em fase sólida headspace combinada com cromatografia gasosa-espectrometria de massa. Em seguida, foram calculados os valores aromáticos característicos, que refletem o grau de percepção olfativa pelo ser humano, de cada VOC na planta de acordo com seus respectivos limiares aromáticos. Os resultados mostraram que três COVs principais medidos do mais alto ao mais baixo são benzoatos de metila, terpenos e derivados de ácidos graxos. Seu conteúdo aumentou após a abertura do botão floral e atingiu o máximo na metade da floração total. Em uma flor, a maioria dos teores de COV foram medidos em pétalas e apenas traços em outros tecidos, como estame, pistilo. Uma pequena quantidade de COVs foi determinada nas folhas também. Todos os valores aromáticos foram quase correspondentes aos teores de três COVs principais, indicando que a fragrância da flor surge verdadeiramente desses componentes de COV. Para entender o mecanismo de formação deste tipo principal de fragrância e sua regulação, primeiramente isolamos um gene de RfBAMT da pétala de Rhododendron fortunei usando clonagem homóloga e tecnologia RACE. O comprimento total de seu cDNA era de 1383 bp, com um quadro de leitura aberto de 1104 bp, codificando um total de 368 aminoácidos. A análise da árvore filogenética mostrou que RfBAMT foi o mais próximo do BSMT de Camellia japonica, pertencente à família das metiltransferases. Em seguida, medimos o nível de expressão de RfBAMT novamente em quatro estágios de desenvolvimento da flor e em diferentes tecidos e folhas de flores. Os resultados mostraram que o nível de expressão deste gene foi altamente correlacionado positivamente com o conteúdo emitido de benzoatos de metila na floração, implicando que RfBAMT desempenha um papel fundamental na formação e regulação de benzoatos de metila em Rhododendron fortune foi clonado e identificado em nosso estudo e seu nível de expressão foi altamente correlacionado positivamente com o conteúdo emitido de benzoatos de metila nas flores e folhas, o que indicou que este gene pode desempenhar um papel importante na regulação da síntese de benzoato de metila em Rhododendron fortunei.

ABSTRACT The response mechanism and interaction patterns of HIF-1α and p53 in animals in an hypoxic environment are crucial for their hypoxic tolerance and adaptation. Many studies have shown that underground rodents have better hypoxic adaptation characteristics. However, the mechanism by which HIF-1α and p53 in underground rodents respond to hypoxic environments compared with in ground rodents remains unclear. Further, whether a synergy between HIF-1α and p53 enables animals tolerate extremely hypoxic environments is unclear. We studied HIF-1α and p53 expression in the brain tissue and cell apoptosis in the hippocampal CA1 region during 6 hours of acute hypoxia (5% oxygen) in Lasiopodomys mandarinus (Milne-Edwards, 1871) and Lasiopodomys brandtii (Radde, 1861), two closely related small rodents with different life characteristics (underground and aboveground, respectively), using a comparative biology method to determine the mechanisms underlying their adaptation to this environment. Our results indicate that HIF-1α and p53 expression is more rapid in L. mandarinus than in L. brandtii under acute hypoxic environments, resulting in a significant synergistic effect in L. mandarinus. Correlation analysis revealed that HIF-1α expression and the apoptotic index of the hippocampal CA1 regions of the brain tissues of L. mandarinus and L. brandtii, both under hypoxia, were significantly negatively and positively correlated, respectively. Long-term existence in underground burrow systems could enable better adaptation to hypoxia in L. mandarinus than in L. brandtii. We speculate that L. mandarinus can quickly eliminate resulting damage via the synergistic effect of p53 and HIF-1α in response to acute hypoxic environments, helping the organism quickly return to a normal state after the stress.

Abstract Objective: This study aims to replicate the phenotype of Ltbp1 knockout mice in zebrafish, and to address the function of LTBP1 in craniofacial development. Methods: Whole mount in situ hybridization (WISH) of ltbp1 was performed at critical periods of zebrafish craniofacial development to explore the spatial-temporal expression pattern. Furthermore, we generated morpholino based knockdown model of ltbp1 to study the craniofacial phenotype. Results: WISH of ltbp1 was mainly detected in the mandibular jaw region, brain trunk, and internal organs such as pancreas and gallbladder. And ltbp1 colocalized with both sox9a and ckma in mandibular region. Morpholino based knockdown of ltbp1 results in severe jaw malformation. Alcian blue staining revealed severe deformity of Meckel's cartilage along with the absence of ceratobranchial. Three-dimension measurements of ltbp1 morphants jaws showed decrease in both mandible length and width and increase in open mouth distance. Expression of cartilage marker sox9a and muscle marker ckma was decreased in ltbp1 morphants. Conclusions: Our experiments found that ltbp1 was expressed in zebrafish mandibular jaw cartilages and the surrounding muscles. The ltbp1 knockdown zebrafish exhibited phenotypes consistent with Ltbp1 knockout mice. And loss of ltbp1 function lead to significant mandibular jaw defects and affect both jaw cartilages and surrounding muscles.

Left atrial diameter (LAD) has been considered an independent risk factor for atrial fibrillation (AF) relapse after pulmonary vein isolation (PVI). However, whether LAD or other factors are more predictive of late recurrence in patients with paroxysmal AF remains unclear. We aimed to evaluate the value of pulmonary vein (PV) parameters for predicting AF relapse 1 year after patients underwent cryoablation for paroxysmal AF. Ninety-seven patients with paroxysmal AF who underwent PVI successfully were included. PV parameters were measured through computed tomography scans prior to PVI. A total of 28 patients had recurrence of AF at one-year follow-up. The impact of several variables on recurrence was evaluated in multivariate analyses. LAD and the time from first diagnosis of AF to ablation maintained its significance in predicting the relapse of AF after relevant adjustments in multivariate analysis. When major diameter of right inferior pulmonary vein (RIPV) (net reclassification improvement (NRI) 0.179, CI=0.031–0.326, P<0.05) and cross-sectional area (CSA) of RIPV (NRI: 0.122, CI=0.004–0.240, P<0.05) entered the AF risk model separately, the added predictive capacity was large. The accuracy of the two parameters in predicting recurrence of AF were not inferior (AUC: 0.665 and 0.659, respectively) to echocardiographic LAD (AUC: 0.663). The inclusion of either RIPV major diameter or CSA of RIPV in the model increased the C-index (0.766 and 0.758, respectively). We concluded that major diameter of RIPV had predictive capacity similar to or even better than that of LAD for predicting AF relapse after cryoablation PVI.

ABSTRACT Folium Eriobotryae, the dried leaves of loquat (Eriobotrya japonica, (Thunb.) Lindl., Rosaceae), is a traditional Chinese medicine used to treat cough with phlegm in China. Fallen and growing loquat leaves were tested for their effect on coughing and expectoration in mice. HPLC-ELSD and HPLC-MS analyses of aqueous and ethanol extracts of fallen or growing leaves were used to identify the chemical components responsible for this effect. Both the aqueous and ethanol extracts of growing and fallen leaves of loquat contained antitussive and expectorant activities. Moreover, an aqueous extract of growing loquat leaves with a higher flavonoid content displayed a stronger expectorant activity while the ethanol extract of fallen loquat leaves that contained a higher content of triterpenoid acids induced a stronger antitussive activity.