Screening and analysis on the differentially expression genes between diploid and autotetraploid watermelon by using of digital gene expression profile / Triagem e análise da expressão diferencial dos genes entre melancia diplóide e autotetraplóide através do perfil de expressão gênica digital

Abstract Synthetic polyploids are key breeding materials for watermelon. Compared with diploid watermelon, the tetraploid watermelon often exhibit wide phenotypic differences and differential gene expression. Digital gene expression (DGE) profile technique was performed in this study to present gene expression patterns in an autotetraploid and its progenitor diploid watermelon, and deferentially expressed genes (DEGs) related to the abiotic and biotic stress were also addressed. Altogether, 4,985 DEGs were obtained in the autotetraploid against its progenitor diploid, and 66.02% DEGs is up-regulated. GO analysis shows that these DEGs mainly distributed in 'metabolic process', 'cell' and 'catalytic activity'. KEGG analysis revealed that these DEGs mainly cover 'metabolic pathways', 'secondary metabolites' and 'ribosome'. Moreover, 134 tolerance related DEGs were identified which cover osmotic adjustment substance, protective enzymes/protein, signaling proteins and pathogenesis-related proteins. This study present the differential expression of stress related genes and global gene expression patterns at background level in autotetraploid watermelons. These new evidences could supplement the molecular theoretical basis for the better resistance after the genome doubling in the gourd family.

Resumo Poliploides sintéticos são materias fundamentais para melhoramento genético da melancia. Comparativamente ao seu homólogo diploide, a melancia tetraploide apresenta amplas diferenças genotípica e fenotípica e diferença de expressão gênica. A expressão gênica digital ou DGE (digital gene expression) foi utilizada neste estudo para representar o perfil de expressão gênica da melancia autotetraploide e seu progenitor diploide e a expressão diferencial de genes relacionados ao estresse biótico e abiótico. Os resultados mostraram que 4.985 DEGs foram observados no organismo autotetraploide, sendo que, deste total, 66.02%foram supra-regulados. A análise de ontologia gênica (GO) mostrou que estes DEGs estão relacionados principalmente com processos metabólicas, célula e atividade catalítica, abrangendo de acordo com a análise de genes e genoma (KEGG) rotas metabólicas, metabolismo secundário e ribossomos. Além disso, 134 genes de defesa foram identificados, abrangendo substâncias de ajuste osmótico, enzimas/proteínas de proteção, proteínas sinalizadoras e proteínas relacionadas à patogênese. Este estudo mostrou a expressão diferencial de genes relacionados ao estresse e o perfil global de expressão gênica de melancia autotetraploide, estes resultados podem complementar, a nível molecular, o entendimento do fator resistência após a duplicação do genoma em cucurbitáceas.

Ginsenoside Rg1 protects human renal tubular epithelial cells from lipopolysaccharide-induced apoptosis and inflammation damage

Ginsenoside Rg1, one of the most notable active components of Panax ginseng, has been widely reported to exert anti-inflammatory actions. This study aimed to reveal whether ginsenoside Rg1 also exhibits beneficial roles against lipopolysaccharide (LPS)-induced apoptosis and inflammation in human renal tubular epithelial cells, and to evaluate the potential role of the component on tubulointerstitial nephritis treatment. HK-2 cells were treated with various doses of ginsenoside Rg1 (0, 50, 100, 150, and 200 μM) in the absence or presence of 5 μg/mL LPS. Thereafter, CCK-8 assay, flow cytometry, western blot, migration assay, reactive oxygen species (ROS) assay, and ELISA were carried out to respectively assess cell viability, apoptosis, migration, ROS activity, and the release of inflammatory cytokines. As a result, ginsenoside Rg1 protected HK-2 cells from LPS-induced injury, as cell viability was increased, cell apoptosis was decreased, and the release of MCP-1, IL-1β, IL-6, and TNF-α was reduced. Ginsenoside Rg1 functioned to HK-2 cells in a dose-dependent manner, and the 150 μM dose exhibited the most protective functions. Ginsenoside Rg1 had no significant impact on cell migration and ROS activity, while it alleviated LPS-induced ROS release and migration impairment. Furthermore, the down-regulations of p-PI3K, p-AKT, and up-regulations of PTEN, p-IκBα, p-p65, Bcl-3 induced by LPS were recovered to some extent after ginsenoside Rg1 treatment. In conclusion, ginsenoside Rg1 protects HK-2 cells against LPS-induced inflammation and apoptosis via activation of the PI3K/AKT pathway and suppression of NF-κB pathway.

Screening and analysis on the differentially expression genes between diploid and autotetraploid watermelon by using of digital gene expression profile

Abstract Synthetic polyploids are key breeding materials for watermelon. Compared with diploid watermelon, the tetraploid watermelon often exhibit wide phenotypic differences and differential gene expression. Digital gene expression (DGE) profile technique was performed in this study to present gene expression patterns in an autotetraploid and its progenitor diploid watermelon, and deferentially expressed genes (DEGs) related to the abiotic and biotic stress were also addressed. Altogether, 4,985 DEGs were obtained in the autotetraploid against its progenitor diploid, and 66.02% DEGs is up-regulated. GO analysis shows that these DEGs mainly distributed in metabolic process, cell and catalytic activity. KEGG analysis revealed that these DEGs mainly cover metabolic pathways, secondary metabolites and ribosome. Moreover, 134 tolerance related DEGs were identified which cover osmotic adjustment substance, protective enzymes/protein, signaling proteins and pathogenesis-related proteins. This study present the differential expression of stress related genes and global gene expression patterns at background level in autotetraploid watermelons. These new evidences could supplement the molecular theoretical basis for the better resistance after the genome doubling in the gourd family.

Resumo Poliploides sintéticos são materias fundamentais para melhoramento genético da melancia. Comparativamente ao seu homólogo diploide, a melancia tetraploide apresenta amplas diferenças genotípica e fenotípica e diferença de expressão gênica. A expressão gênica digital ou DGE (digital gene expression) foi utilizada neste estudo para representar o perfil de expressão gênica da melancia autotetraploide e seu progenitor diploide e a expressão diferencial de genes relacionados ao estresse biótico e abiótico. Os resultados mostraram que 4.985 DEGs foram observados no organismo autotetraploide, sendo que, deste total, 66.02%foram supra-regulados. A análise de ontologia gênica (GO) mostrou que estes DEGs estão relacionados principalmente com processos metabólicas, célula e atividade catalítica, abrangendo de acordo com a análise de genes e genoma (KEGG) rotas metabólicas, metabolismo secundário e ribossomos. Além disso, 134 genes de defesa foram identificados, abrangendo substâncias de ajuste osmótico, enzimas/proteínas de proteção, proteínas sinalizadoras e proteínas relacionadas à patogênese. Este estudo mostrou a expressão diferencial de genes relacionados ao estresse e o perfil global de expressão gênica de melancia autotetraploide, estes resultados podem complementar, a nível molecular, o entendimento do fator resistência após a duplicação do genoma em cucurbitáceas.

Fetal development of regulatory mechanisms for body fluid homeostasis

The balance of body fluids is critical to health and the development of diseases. Although quite a few review papers have shown that several mechanisms, including hormonal and behavioral regulation, play an important role in body fluid homeostasis in adults, there is limited information on the development of regulatory mechanisms for fetal body fluid balance. Hormonal, renal, and behavioral control of body fluids function to some extent in utero. Hormonal mechanisms including the renin-angiotensin system, aldosterone, and vasopressin are involved in modifying fetal renal excretion, reabsorption of sodium and water, and regulation of vascular volume. In utero behavioral changes, such as fetal swallowing, have been suggested to be early functional development in response to dipsogens. Since diseases, such as hypertension, can be traced to fetal origin, it is important to understand the development of fetal regulatory mechanisms for body fluid homeostasis in this early stage of life. This review focuses on fetal hormonal, behavioral, and renal development related to regulation of body fluids in utero.

Effect of supplementation of green tea polyphenols on the developmental competence of bovine oocytes in vitro

The objective of the present study was to examine the effect of green tea polyphenols (GTPs) supplementation during in vitro maturation, in vitro fertilization, and in vitro culture on the developmental competence of bovine oocytes. Cumulus-oocyte complexes aspirated from the ovaries were matured in vitro (38.5°C for 24 h) and fertilized (38.5°C for 15-18 h) and embryos were cultured (38.5°C for 192 h) in a defined conditioned medium with or without GTPs supplementation. The GTPs used in the present study contained 99 percent catechin derivatives, with the major components being 50 percent (-)-epigallocatechin gallate, 22 percent (-)-epicatechin gallate, 18 percent (-)-epigallocatechin, and 10 percent (-)-epicatechin. Four replicate trials were done for each type of experiment. GTPs supplementation (15 æM) of the maturation medium led to a significant increase in the rate of blastocyst formation (34.0 vs 21.4 percent, P < 0.05). However, the rate of blastocyst formation was not improved when higher GTPs concentrations (20 or 25 æM) were added to the in vitro maturation medium. During in vitro fertilization, supplementation with higher GTPs concentrations (20 or 25 æM) significantly reduced the rate of blastocyst formation (P < 0.05). Supplementation of the culture medium with 15 æM GTPs improved the rate of blastocyst formation, while higher GTPs concentrations (25 æM) significantly reduced embryo development (P < 0.05). In conclusion, these results demonstrate that supplementation with GTPs at low concentration (15 æM) during in vitro maturation and in vitro culture improved the developmental competence of bovine oocytes.

Development of fetal nicotine and muscarinic receptors in utero

The role of acetylcholine in the central and peripheral nervous systems is well established in adults. Cholinergic modulation of vascular functions and body fluid balance has been extensively studied. In the embryo-fetus, cholinergic receptors are widespread in the peripheral and central systems, including smooth muscle and the epithelial lining of the cardiovascular, digestive, and urinary systems, as well as in the brain. Fetal nicotine and muscarinic receptors develop in a pattern (e.g., amount and distribution) related to gestational periods. Cholinergic mechanisms have been found to be relatively intact and functional in the control of vascular homeostasis during fetal life in utero at least during the last third of gestation. This review focuses on the development of fetal nicotine and muscarinic receptors, and provides information indicating that central cholinergic systems are well developed in the control of fetal blood pressure and body fluid balance before birth. Therefore, the development of cholinergic systems in utero plays an important role in fetal vascular regulation, gastrointestinal motility, and urinary control.

Experimental studies on small hooks preceding large hooks in the growth and development of Taenia solium metacestodes.

In the present study, we have determined the growth and development pattern of rostellar hooklets of Taenia solium cysticerci (Zhengzhou and Harbin strains) in three pigs (1 SEM and 2 L-SEM strains) 89-196 days post experimental infection. A total of 3,675 cysticerci were collected from 3 pigs, 3,007 (82%) of 3,675 cysticerci were evaginated by enzyme method. 439 (15%) evaginated cysticerci were carefully examined and measured after dehydration, staining, and mounting on microscopic slides. Among 439 cysticerci, 234 (53%) had pair rostellar hooks, 88 (20%) with unpair hooks, 60 (14%) only small (outer row) hooks, and 57 (13%) no hooks including 34 hooks were completely dropped and 23 no hooks developed. The number ranged from 10 to 17 pairs for pair hooks and 1 to 29 for unpair ones. The length and width of rostallar hooks on the scolex of cysticerci were usually larger in the pig with longer infection time. Moreover, cysticerci with pair and unpair rostellar hooks had only small hooks and no hooks were present on their scolices. However, cysticerci with only large (inner row) hooks were not found. These findings indicate that the growth and development of small hooks precedes that of the large hooks in the formation of the two-row pattern rostellar hook in Cysticercus cellulosae.

Studies on abnormality of metacestodes and adult worms of Taenia solium and Taenia saginata asiatica in rodents and pigs.

Abnormalities are not uncommon in Taenia saginata and T. solium. After examining 328 mature proglottids from 2 adult worms from two experimentally infected hamsters, 13 (4.0%) were found to have no genital pore but with numerous testes and several vas efferents; 1 (0.3%) one genital pore with one reproductive system; 12 (3.7%) one on each side with two sets of reproductive system; 17 (5.2%) two on one side with 2 sets of reproductive system; 8 (2.4%) one on one side and two on the other side with 3 sets of reproductive system; 2 (0.6%) two on each side with 4 sets of reproductive system; 4 (1.2%) three on one side with 3 sets of reproductive system, and 4 one on one side and three on the other side with 4 sets of reproductive system. Nine evaginated abnormal cysticerci of T. s. asiatica from three experimentally infected SCID mice each had two protoscoleces and a big bladder. From two experimentally infected pigs, one abnormal cysticercus was observed to have two invaginated canals each in one end. Another one had a neck-band behind the scolex and a big bladder. This paper is not only the first report of abnormality of T. solium from hamster but also the first one of abnormal cysticerci of T. s. asiatica from pigs and mice.

Studies on clinical manifestations, diagnosis and control of paragonimiasis in China.

Paragonimiasis is a common parasitic zoonosis in China. Its clinical manifestations may be classified into two clinical entities, eg, paragonimiasis westermani type and paragonimiasis szechuanensis type. The latter mainly manifests as trematode larva migrans. The immunodiagnostic methods frequently used in the diagnosis of paragonimiasis are intradermal test, CFT and ELISA. The detection of circulating antigens in patients has been used in China for diagnosis and bithionol and praziquantel are drugs of choice in the treatment of paragonimiasis. Combined measures including mass survey, mass treatment and health education have been found to be effective in the control of paragonimiasis in China.