The maize transcription factor CCT regulates drought tolerance by interacting with Fra a 1, E3 ligase WIPF2, and auxin response factor Aux/IAA8.

Plants are commonly exposed to abiotic stressors, which can affect their growth, productivity, and quality. Previously, the maize transcription factor ZmCCT was shown to be involved in the photoperiod response, delayed flowering, and quantitative resistance to Gibberella stalk rot. In this study, we demonstrate that ZmCCT can regulate plant responses to drought. ZmCCT physically interacted with ZmFra a 1, ZmWIPF2, and ZmAux/IAA8, which localized to the cell membrane, cytoplasm, and nucleus, respectively, both in vitro and in vivo in a yeast two-hybrid screen in response to abiotic stress. Notably, ZmCCT recruits ZmWIPF2 to the nucleus, which has strong E3 self-ubiquitination activity dependent on its RING-H2 finger domain in vitro. When treated with higher indole-3-acetic acid/abscisic acid ratios, the height and root length of Y331-ΔTE maize plants increased. Y331-ΔTE plants exhibited increased responses to exogenously applied auxin or ABA compared to Y331 plants, indicating that ZmCCT may be a negative regulator of ABA signalling in maize. In vivo, ZmCCT promoted indole-3-acetic acid biosynthesis in ZmCCT-overexpressing Arabidopsis. RNA-sequencing and DNA affinity purification-sequencing analyses showed that ZmCCT can regulate the expression of ZmRD17, ZmAFP3, ZmPP2C, and ZmARR16 under drought. Our findings provide a detailed overview of the molecular mechanism controlling ZmCCT functions and highlight that ZmCCT has multiple roles in promoting abiotic stress tolerance.

AtCNGC2 is involved in jasmonic acid-induced calcium mobilization.

Calcium (Ca(2+)) mobilization is a central theme in various plant signal transduction pathways. We demonstrate that Arabidopsis thaliana cyclic nucleotide-gated channel 2 (AtCNGC2) is involved in jasmonic acid (JA)-induced apoplastic Ca(2+) influx in Arabidopsis epidermal cells. Ca(2+) imaging results showed that JA can induce an elevation in the cytosolic cAMP concentration ([cAMP]cyt), reaching a maximum within 3 min. Dibutyryl cAMP (db-cAMP), a cell membrane-permeable analogue of cAMP, induced an increase in the cytosolic Ca(2+) concentration ([Ca(2+)]cyt), with a peak at 4 min. This [Ca(2+)]cyt increase was triggered by the JA-induced increase in [cAMP]cyt. W-7[N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide], an antagonist of calmodulin, positively modulated the JA-induced increase in [Ca(2+)]cyt, while W-5[N-(6-aminohexyl)-1-naphthalenesulfonamide], an inactive antagonist of calmodulin, had no apparent effect. db-cAMP and JA positively induced the expression of primary (i.e. JAZ1 and MYC2) and secondary (i.e. VSP1) response genes in the JA signalling pathway in wild-type Arabidopsis thaliana, whereas they had no significant effect in the AtCNGC2 mutant 'defense, no death (dnd1) plants. These data provide evidence that JA first induces the elevation of cAMP, and cAMP, as an activating ligand, activates the AtCNGC2 channel, resulting in apoplastic Ca(2+) influx through AtCNGC2.

Identification of 7 stress-related NAC transcription factor members in maize (Zea mays L.) and characterization of the expression pattern of these genes.

NAC proteins are plant-specific transcription factors that play essential roles in plant development and various abiotic stress responses. A comprehensive analysis of maize NAC genes was performed in this study. A total of 157 non-redundant maize NAC genes including seven membrane-bound members were identified and found to be unevenly distributed on 10 maize chromosomes. Motif composition analysis indicated that the maize NAC proteins share three relatively conserved motifs in the NAC domain within the N-terminal region. Phylogenetic analysis of 157 maize NAC proteins accompanied by 117 NAC proteins from Arabidopsis and 151 from rice were presented. The NAC proteins evaluated were divided into two large groups including 18 subgroups. Gene duplication analysis indicated that gene loss occurred during maize evolution. Seven NAC members that belong to the same clade of maize NAC domain genes were isolated, and overlapping expression patterns were observed under various abiotic stresses, including low temperature, high salinity and dehydration, and phytohormone abscisic acid treatments. This suggested that NAC members function as stress-responsive transcription factors in ABA-dependent signaling pathways. Relatively higher expression levels of these selected maize NAC genes were detected in roots. The stress responsive NAC genes may have applications in molecular breeding to improve crop stress tolerance.

Human Parathyroid Hormone (1-34) accelerates skin wound healing through inducing cell migration via up-regulating the expression of Rac1.

Delayed wound healing is a public issue that imposes a significant burden on both society and the patients themselves. To date, although numerous methods have been developed to accelerate the speed of wound closure, the therapeutic effects are partially limited due to the complex procedures, high costs, potential side effects, and ethical concerns. While some studies have reported that the in-vivo application of Human Parathyroid Hormone (1-34) (hPTH(1-34)) promotes the wound-healing process, the definitive role and underlying mechanisms through which it regulates the behavior of fibroblasts and keratinocytes remains unclear. Herein, hPTH(1-34)'s role in cell migration is evaluated with a series of in-vitro and in-vivo studies, whereby hPTH(1-34)'s underlying mechanism in activating the two types of cells was detected. The in-vitro study revealed that hPTH(1-34) enhanced the migration of both fibroblasts and HaCaT cells. Ras-associated C3 botulinum toxin subunit 1 (Rac1), a classical member of the Rho family, was upregulated in hPTH(1-34)-treated fibroblasts and HaCaT cells. Further study by silencing the expression of Rac1 with siRNA reversed the hPTH(1-34)-enhanced cell migration, thus confirming that Rac1 was involved in hPTH(1-34)-induced cell behavior. In-vivo study on rat wound models confirmed the effects of hPTH(1-34) on fibroblasts and keratinocytes, with increased collagen deposition, fibroblasts accumulation, and Rac1 expression in the hPTH(1-34)-treated wounds. In summary, the present study demonstrated that hPTH(1-34) accelerated wound healing through enhancing the migration of cells through the up-regulation of Rac1 expression.

Transcriptome analysis in contrasting maize inbred lines and functional analysis of five maize NAC genes under drought stress treatment.

Drought substantially influences crop growth and development. NAC (NAM, ATAF1/2, and CUC2) transcription factors (TFs) have received much attention for their critical roles in drought stress responses. To explore the maize NAC genes in response to drought stress, the transcriptome sequencing data of NAC TFs in two maize inbred lines, the drought tolerance line H082183 and the sensitive line Lv28, were used to screen the differentially expressed genes (DEGs). There were 129 maize NAC protein-coding genes identified, of which 15 and 20 NAC genes were differentially expressed between the two genotypes under MD and SD treatments, respectively. Meanwhile, the phylogenetic relationship of 152 non-redundant NAC family TFs in maize was generated. The maize NAC family proteins were grouped into 13 distinct subfamilies. Five drought stress-responsive NAC family members, which were designed as ZmNAP, ZmNAC19, ZmNAC4, ZmJUB1(JUBGBRUNNEN1), and ZmNAC87, were selected for further study. The expression of ZmNAP, ZmNAC19, ZmNAC4, ZmJUB1, and ZmNAC87 were significantly induced by drought, dehydration, polyethylene glycol (PEG) stress, and abscisic acid (ABA) treatments. The overexpressing Arabidopsis of these five NAC genes was generated for functional characterization, respectively. Under different concentrations of NaCl, D-mannitol stress, and ABA treatments, the sensitivity of ZmNAP-, ZmNAC19-, ZmNAC4-, ZmJUB1-, and ZmNAC87-overexpressing lines was significantly increased at the germination stage compared to the wild-type lines. The overexpression of these five NAC members significantly improved the drought stress tolerance in transgenic Arabidopsis. Yeast two-hybrid screening analysis revealed that ZmNAP may cooperatively interact with 11 proteins including ZmNAC19 to activate the drought stress response. The above results inferred that ZmNAP, ZmNAC19, ZmNAC4, ZmJUB1, and ZmNAC87 may play important roles in the plant response to drought stress and may be useful in bioengineering breeding and drought tolerance improvement.

[Debridement and bone grafting with internal fixation via anterior approach for the treatment of tuberculosis of lower cervical vertebrae].

OBJECTIVE: To evaluate the clinical effects of debridement and bone grafting with internal fixation via anterior approach in treatment of tuberculosis of lower cervical vertebrae. METHODS: The clinical data of 15 patients with tuberculosis of lower cervical vertebrae who accepted the treatment of one-stage debridement and bone grafting with internal fixation from June 2010 to December 2018 were retrospectively analyzed. There were 9 males and 6 females, aged from 39 to 72 years with an average of (54.67±10.75) years. The lesion segment was C4 to C6. Pre- and post-operative neurologic functions were evaluated by ASIA grade. All the patients underwent the X-ray films of positive and lateral of cervical spine before and after the operation and accepted the periodic review of CT to evaluate the bone grafting. RESULTS: All the 15 operations were successful, no neurological or vascular injury occurred during the operation, and all patients were followed up for 18 to 52 months. The clinical symptoms improved significantly during the follow-up period and CT showed good bone grafting fusion. One patient suffered a relapse of the illness 3 years later, but was healed during the follow-up visit by strengthening the anti tuberculosis therapy. CONCLUSION: For the patients with vertebral destruction and loss of cervical stability, one-stage debridement and bone grafting with internal fixation via anterior approach has definite curative effects. On the basis of standard anti tuberculosis treatment before operation, the long-term standard anti-tuberculosis treatment after operation is the key to healing the tuberculosis of lower cervical vertebrae.

[Application of bone flap pedicled on retrograde branch of radial artery for treatment of old scaphoid bone fractures of type AO-B].

OBJECTIVE: To investigate application of the bone flap pedicled on the retrograde branch of radial artery for treatment of old scaphoid bone fractures of type AO-B. METHODS: From October 2007 to October 2011,41 patients with old scaphoid bone fractures of type AO-B were treated by transplantation of the bone flap pedicled on the retrograde branch of radial artery including 26 males and 15 females with an average of (27.3±4.5) years old ranging from 16 to 43 years old. The courses before operation ranged from 6 to 22 months with an average of 11 months. All fractures belonged to the type B of AO classification, that is old wrist fracture of scaphoid bone. All patients' wrist function (pain, function, motion, grip strength) were evaluated by Cooney's modifiedwrist scoring system before and 6 months after operation,and the conditions of bone healing were observed during the follow-up time. RESULTS: Among them, 36 patients were followed up from 4 to 15 months with an average of 8.3 months. The wounds were healed well without other complications as infection appearing. X-rays or CT confirmed that all fractures were healed completely. The Cooney wrist score was improved from preoperative 53.61±13.97 to postoperative 81.81±8.71 (P<0.01). CONCLUSION: The operation of transplantation of the bone flap pedicled on the retrograde branch of radial artery is an effective method to treat old scaphoid bone fractures,which is scientific and has curative effects, and valuable for clinical application.

Bioinformatic identification and experimental validation of miRNAs from foxtail millet (Setaria italica).

MiRNAs are a novel group of non-coding small RNAs that negatively regulate gene expression. Many miRNAs have been identified and investigated extensively in plant species with sequenced genomes. However, few miRNAs have been identified in foxtail millet (Setaria italica), which is an ancient cereal crop of great importance for dry land agriculture. In this study, 271 foxtail millet miRNAs belonging to 44 families were identified using a bioinformatics approach. Twenty-three pairs of sense/antisense miRNAs belonging to 13 families, and 18 miRNA clusters containing members of 8 families were discovered in foxtail millet. We identified 432 potential targets for 38 miRNA families, most of which were predicted to be involved in plant development, signal transduction, metabolic pathways, disease resistance, and environmental stress responses. Gene ontology (GO) analysis revealed that 101, 56, and 23 target genes were involved in molecular functions, biological processes, and cellular components, respectively. We investigated the expression patterns of 43 selected miRNAs using qRT-PCR analysis. All of the miRNAs were expressed ubiquitously with many exhibiting different expression levels in different tissues. We validated five predicted targets of four miRNAs using the RNA ligase mediated rapid amplification of cDNA end (5'-RLM-RACE) method.

[Reconstruction of calcaneal thalamus and subtalar arthrodesis to treat old antiquated intra-articular calcaneal fractures of Sanders type III].

OBJECTIVE: To observe the outcome of treatment for serious old intra-articular calcaneal fracture by reconstruction of calcaneal thalamus and subtalar arthrodesis and to summarize the operative indications and its attention points. METHODS: From March 2006 to July 2011,26 patients with Sanders type III old intra-articular calcaneal fracture were treated including 15 males and 11 females with an average age of 34 years old ranging from 27 to 45 years old. The clinical courses ranged from 7 to 24 months with an average of 18 months. Before the operation,X-ray and CT showed that Gissane angle increased while Böhler angle decreased,and calcaneus broadened,bulging on both sides. After the operations,all patients tested by imaging examination, and the width of calcaneus, the height of calcaneal thalamus, the Böhler angle and Gissane angle were measured to compare with the preoperative data. Besides, for identification of improvement of the operation,the functions of patient's feet after the operation were graded according to AOFAS Ankle-Hindfoot Scale to compare with preoperative data. RESULTS: Among them, 24 patients were followed-up from 5 to 26 months with an average of 19 months. X-rays confirmed that all cases healed successfully. There were no serious infection, and only one skin necrosis case. Sural neurocutaneous island flap was used to repair the skin with success. According to AOFAS Ankle-Hindfoot Scale, the total score increased from preoperative (41.00 +/- 8.22) to postoperative (79.04 +/- 7.46). There were 3 cases of excellent result, 15 cases of good result,and 6 cases of fair result. Postoperative width of calcaneus, the height of thalamus, Böhler angle and Gissane angle were better than that of preoperative data, and had statistical significance between two groups. CONCLUSION: Subtalar arthrodesis with the reconstruction of calcaneal thalamus is an effective way to treat old intra-articular calcaneal fractures. It can correct the calcaneal deformity, restore the shape of foot and improve the function of hind foot.

Influx of extracellular Ca2+ involved in jasmonic-acid-induced elevation of [Ca2+]cyt and JR1 expression in Arabidopsis thaliana.

The changes in cytosolic Ca2+ levels play important roles in the signal transduction pathways of many environmental and developmental stimuli in plants and animals. We demonstrated that the increase in cytosolic free Ca2+ concentration ([Ca2+]cyt) of Arabidopsis thaliana leaf cells was induced by exogenous application of jasmonic acid (JA). The elevation of [Ca2+]cyt was detected within 1 min after JA treatment by the fluorescence intensity using laser scanning confocal microscopy, and the elevated level of fluorescence was maintained during measuring time. With pretreatment of nifedipine (Nif), a nonpermeable L-type channel blocker, the fluorescence of [Ca2+]cyt induced by JA was inhibited in a dose-dependent manner. In contrast, verapamil, another L-type channel blocker, had no significant effect. Furthermore, Nif repressed JA-induced gene expression of JR1 but verapamil did not. JA-induced gene expression could be mimicked by higher concentration of extracellular Ca2+. W-7 [N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide], an antagonist of calmodulin (CaM), blocked the JA induction of JR1 expression while W-5 [N-(6-aminohexyl)-1-naphthalenesulfonamide], its inactive antagonist, had no apparent effect. These data provide the evidence that the influx of extracellular Ca2+ through Nif sensitive plasma membrane Ca2+ channel may be responsible for JA-induced elevation of [Ca2+]cyt and downstream gene expression, CaM may be also involved in JA signaling pathway.