High throughput sequencing of herbaceous peony small RNAs to screen thermo-tolerance related microRNAs

Zhaojun Hao,Ding Liu,Saijie Gong,Daqiu Zhao,Jun Tao 한국유전학회 2017 Genes & Genomics Vol.39 No.4

Herbaceous peony (Paeonia lactiflora Pall.) is an ornamental flower with wide applications for landscape. However, its application potential is greatly restricted because it is prone to heat stress injury. Although there are some studies about thermo-tolerance of P. lactiflora which focused on physiological and transcriptional level, the information on post-transcriptional level is lacked. Here, we chose a thermo-tolerant cultivar ‘Zifengyu’ and a moderately thermo-tolerant cultivar ‘Hongyanzhenghui’ under the natural long summer heat to investigate heat responsive miRNAs by miRNA sequencing. A total of 24,008,974 and 21,981,360 raw reads were generated from ‘Zifengyu’ and ‘Hongyanzhenghui’, respectively, as well as 45,016,796 total sRNAs and 6,297,111 unique sRNAs were obtained. Then 271 conserved and 9 novel miRNAs were identified in ‘Zifengyu’, while 236 conserved and 11 novel miRNAs were identified in ‘Hongyanzhenghui’. Among them, 71 known miRNAs with 272 potential target genes and 3 novel miRNAs with 4 potential target genes were significantly differentially expressed. Expression analysis of the 7 candidate miRNAs previously reported to be regulated by heat stress and their target genes were performed. It showed that miR172c-3p, miR395a, miR397a, miR408-5p and miR827 were up-regulated by heat stress, and expressed much higher in thermo-tolerant cultivar ‘Zifengyu’, suggesting that they might be involved in heat stress response at the post-transcriptional level. These results could provide a better understanding about the thermos-tolerant mechanism of P. lactiflora on post-transcriptional level.

The bifunctional effect of propofol on thromboxane agonist (U46619)-induced vasoconstriction in isolated human pulmonary artery

Hao, Ning,Wang, Zhaojun,Kuang, Sujuan,Zhang, Guangyan,Deng, Chunyu,Ma, Jue,Cui, Jianxiu The Korean Society of Pharmacology 2017 The Korean Journal of Physiology & Pharmacology Vol.21 No.6

Propofol is known to cause vasorelaxation of several systemic vascular beds. However, its effect on the pulmonary vasculature remains controversial. In the present study, we investigated the effects of propofol on human pulmonary arteries obtained from patients who had undergone surgery. Arterial rings were mounted in a Multi-Myograph system for measurement of isometric forces. U46619 was used to induce sustained contraction of the intrapulmonary arteries, and propofol was then applied (in increments from $10-300{\mu}m$). Arteries denuded of endothelium, preincubated or not with indomethacin, were used to investigate the effects of propofol on isolated arteries. Propofol exhibited a bifunctional effect on isolated human pulmonary arteries contracted by U46619, evoking constriction at low concentrations ($10-100{\mu}m$) followed by secondary relaxation (at $100-300{\mu}m$). The extent of constriction induced by propofol was higher in an endothelium-denuded group than in an endothelium-intact group. Preincubation with indomethacin abolished constriction and potentiated relaxation. The maximal relaxation was greater in the endothelium-intact than the endothelium-denuded group. Propofol also suppressed $CaCl_2$-induced constriction in the 60 mM $K^+$-containing $Ca^{2+}$-free solution in a dose-dependent manner. Fluorescent imaging of $Ca^{2+}$ using fluo-4 showed that a 10 min incubation with propofol ($10-300{\mu}m$) inhibited the $Ca^{2+}$ influx into human pulmonary arterial smooth muscle cells induced by a 60 mM $K^+$-containing $Ca^{2+}$-free solution. In conclusion, propofol-induced arterial constriction appears to involve prostaglandin production by cyclooxygenase in pulmonary artery smooth muscle cells and the relaxation depends in part on endothelial function, principally on the inhibition of calcium influx through L-type voltage-operated calcium channels.

The bifunctional effect of propofol on thromboxane agonist (U46619)-induced vasoconstriction in isolated human pulmonary artery

Ning Hao,Zhaojun Wang,Sujuan Kuang,Guangyan Zhang,Chunyu Deng,Jue Ma,Jianxiu Cui 대한생리학회-대한약리학회 2017 The Korean Journal of Physiology & Pharmacology Vol.13 No.3

Propofol is known to cause vasorelaxation of several systemic vascular beds. However, its effect on the pulmonary vasculature remains controversial. In the present study, we investigated the effects of propofol on human pulmonary arteries obtained from patients who had undergone surgery. Arterial rings were mounted in a Multi-Myograph system for measurement of isometric forces. U46619 was used to induce sustained contraction of the intrapulmonary arteries, and propofol was then applied (in increments from 10-300 μM). Arteries denuded of endothelium, preincubated or not with indomethacin, were used to investigate the effects of propofol on isolated arteries. Propofol exhibited a bifunctional effect on isolated human pulmonary arteries contracted by U46619, evoking constriction at low concentrations (10-100 μM) followed by secondary relaxation (at 100-300 μM). The extent of constriction induced by propofol was higher in an endothelium-denuded group than in an endothelium-intact group. Preincubation with indomethacin abolished constriction and potentiated relaxation. The maximal relaxation was greater in the endothelium-intact than the endothelium-denuded group. Propofol also suppressed CaCl₂-induced constriction in the 60 mM K⁺-containing Ca²⁺-free solution in a dose-dependent manner. Fluorescent imaging of Ca²⁺ using fluo-4 showed that a 10 min incubation with propofol (10-300 μM) inhibited the Ca²⁺ influx into human pulmonary arterial smooth muscle cells induced by a 60 mM K⁺-containing Ca²⁺-free solution. In conclusion, propofol-induced arterial constriction appears to involve prostaglandin production by cyclooxygenase in pulmonary artery smooth muscle cells and the relaxation depends in part on endothelial function, principally on the inhibition of calcium influx through L-type voltage-operated calcium channels.

Transcriptome and digital gene expression analysis of herbaceous peony (Paeonia lactiflora Pall.) to screen thermo-tolerant related differently expressed genes

Jun Tao,Zhaojun Hao,Mengran Wei,Saijie Gong,Daqiu Zhao 한국유전학회 2016 Genes & Genomics Vol.38 No.12

Herbaceous peony (Paeonia lactiflora Pall.) is easily injured by heat stress (HS), which greatly restricts its application and promotion. In this study, the thermo-tolerance of three representative P. lactiflora cultivars had been firstly assessed. ‘Zifengyu’ was identified as the thermo-tolerant cultivar with relatively lower values and smaller variations in malondialdehyde, hydrogen peroxide (H2O2) and proline contents under HS. Subsequently, their transcriptomes were sequenced by RNA sequencing (RNAseq) technology to construct a complete database. 81,599 unigenes were obtained, and 34,940 unigenes had been annotated. Moreover, through digital gene expression analysis of thermo-tolerant ‘Zifengyu’ and moderately thermo-tolerant ‘Hongyanzhenghui’, 161 heat stress response genes had been screened involving heat shock protein genes, plant hormone signal transduction related genes, fatty acid synthesis genes, reactive oxygen speciesscavenging genes and secondary metabolites related genes. And the effectively and timely response of these genes to HS could endow thermo-tolerance to ‘Zifengyu’. Among these genes, 11 key thermo-tolerant related genes whose expressions were all significantly up-regulated in ‘Zifengyu’ and ‘Hongyanzhenghui’ during development and the former possessed higher levels could be regarded as the candidate genes, including isoprene synthase gene, 2 peroxidase genes, 3-oxoacyl-acyl carrier protein reductase gene (FabG), 3 transcription factor genes (bHLH, NAC and WRKY), HSP20 and 3 HSP70. These results could provide a better understanding of heat stress response in P. lactiflora, and pave for the breeding of thermo-tolerant cultivars.

Green algae dominance quickly switches to cyanobacteria dominance after nutrient enrichment in greenhouse with high temperature

Wang, Xiaodong,Liu, Xingguo,Qin, Boqiang,Gu, Zhaojun,Wu, Zongfan,Xu, Hao,Zhu, Hao,Cheng, Guofeng,Liu, Huang The Ecological Society of Korea 2015 Journal of Ecology and Environment Vol.38 No.3

In order to understand the mechanisms of conversion between different algal dominance, an experiment was performed in a greenhouse from 22 June to 10 July 2011. The experiment included a treatment group subjected to three instances of nutrient enrichment and a control with no nutrient enrichment. The initial water was dominated by Ankistrodesmus of Chlorophyta. The average water temperature at 08:30 h and 14:00 h during the experiment was $31.6^{\circ}C$ and $34.6^{\circ}C$, respectively. The results showed that the total nitrogen (TN), total phosphorus (TP), dissolved total nitrogen (DTN), dissolved total phosphorus (DTP), and soluble reactive phosphorus (SRP) concentrations in the treatment were significantly higher than in the control (P < 0.05). However, the TN/TP and DTN/DTP in the control was higher than in the treatment (P < 0.05). The dominant algae in the control did not change during the experiment, while the dominant algae in the treatment switched to Planktothrix of Cyanophyta on day 9. The chlorophyll a (Chl-a), wet weight of all algae, wet weight of Cyanophyta, and percentage of Cyanophyta in the control were all significantly lower than in the treatment (P < 0.05). Amounts of zooplankton, especially rotifers, were present at the end of the experimental period. The density of rotifers between the control and treatment was not significantly different (P > 0.05), while the copepod density in the treatment was higher than in the control (P < 0.05). We conclude that green algae dominance quickly switches to cyanobacteria dominance after nutrient enrichment in a greenhouse with elevated temperature.

Green algae dominance quickly switches to cyanobacteria dominance after nutrient enrichment in greenhouse with high temperature

Xiaodong Wang,Xingguo Liu1,Boqiang Qin,Zhaojun Gu,Hao Xu,Hao Zhu,Guofeng Cheng,Huang Liu 한국생태학회 2015 Journal of Ecology and Environment Vol.38 No.3

In order to understand the mechanisms of conversion between different algal dominance, an experiment was performed in a greenhouse from 22 June to 10 July 2011. The experiment included a treatment group subjected to three instances of nutrient enrichment and a control with no nutrient enrichment. The initial water was dominated by Ankistrodesmus of Chlorophyta. The average water temperature at 08:30 h and 14:00 h during the experiment was 31.6°C and 34.6°C, respec¬tively. The results showed that the total nitrogen (TN), total phosphorus (TP), dissolved total nitrogen (DTN), dissolved total phosphorus (DTP), and soluble reactive phosphorus (SRP) concentrations in the treatment were significantly higher than in the control (P < 0.05). However, the TN/TP and DTN/DTP in the control was higher than in the treatment (P < 0.05). The dominant algae in the control did not change during the experiment, while the dominant algae in the treat¬ment switched to Planktothrix of Cyanophyta on day 9. The chlorophyll a (Chl-a), wet weight of all algae, wet weight of Cyanophyta, and percentage of Cyanophyta in the control were all significantly lower than in the treatment (P < 0.05). Amounts of zooplankton, especially rotifers, were present at the end of the experimental period. The density of rotifers between the control and treatment was not significantly different (P > 0.05), while the copepod density in the treatment was higher than in the control (P < 0.05). We conclude that green algae dominance quickly switches to cyanobacteria dominance after nutrient enrichment in a greenhouse with elevated temperature

Effects of a mild heat treatment on mouse testicular gene expression and sperm quality

Zhao, Jun,Zhang, Ying,Hao, Linlin,Wang, Jia,Zhang, Jiabao,Liu, Songcai,Ren, Bingzhong The Korean Society for Integrative Biology 2010 Animal cells and systems Vol.14 No.4

The decrease in sperm quality under heat stress causes a great loss in animal husbandry production. In order to reveal the mechanism underlying the sperm quality decrease caused by heat stress, we first established a mild heat-treated mouse model. Then, the sperm quality was identified. Further, the testicular proteome profile was mapped and compared with the control using 2D electrophoresis and mass spectrometry. Finally, the differential expressed proteins involved in the heat stress response were identified by real-time PCR and Western blotting. The results showed that heat stress caused a significant reduction in mouse sperm quality (P<0.05). Further, 52 protein spots on the 2D gel were found to differ between the heat-shocked tissues and the control. Of these spots, some repair proteins which might provide some explanation for the influence on sperm quality were found. We then focused on Bag-1, Hsp40, Hsp60 and Hsp70, which were found to be differently expressed after heat shock (P<0.05). Further analysis in this heat-shocked model suggests numerous potential mechanisms for heat shock-induced spermatogenic disorders.