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Two homologous genes, Brassica campestris Male Fertility 23a (BcMF23a) and Brassica campestris Male Fertility 23b (BcMF23b), encoding putative pectin methylesterases (PMEs) were isolated from Brassica campestris ssp. chinensis (syn. Brassica rapa ssp. chinensis). These two genes sharing high sequence identity with each other were highly expressed in the fertile flower buds but silenced in the sterile ones of genic male sterile line system ('Bcajh97-01A/B'). Results of RT-PCR and in situ hybridization suggested that BcMF23a and BcMF23b were pollen-expressed genes, whose transcripts were first detected at the binucleate pollen and maintained throughout to the mature pollen grains. Western blot indicated that both of the putative BcMF23a and BcMF23b proteins are approximately 40 kDa, which exhibited extracellular localization revealed by transient expression analysis in the onion epidermal cells. The promoter of BcMF23a was active specifically in pollen during the late pollen developmental stages, while, in addition to the pollen, BcMF23b promoter drove an extra gene expression in the valve margins, abscission layer at the base of the first true leaves, taproot and lateral roots in seedlings.

Taking advantage of the abundant redox active C-type cytochromes on their outer membrane (OMCs), the genus Shewanella enables extracellular electron transfer (EET), which is applicable in various bioelectrical devices. For practical applications, high efficient EET is always desirable. Here, we reveal that tailoring the surface wettability of electrodes can drastically alter the EET activity of microbial Shewanella loihica PV-4: the EET current on a superhydrophilic electrode is over 3 and 10 times higher than that on normal hydrophilic and hydrophobic electrodes, respectively. Worth noting is that a cell suspension with a rather small initial cell density was preferentially used, which can exclude the influence of other unfavorable factors in such a dynamic and flexible living system. It is proposed that a hydrophilic electrode favors a reduced state of OMCs, and consequently both the EET activity and cell proliferation are highly facilitated.

Angiomotin (Amot) is a multifunctional protein involved in endothelial cell migration and tube formation and angiogenesis. However, the biological role and molecular mechanism for the abnormal expression of Amot in breast cancer is poorly understood. The aim of the present study was to examine the function of and relationship between Amot and the Hippo-Yes-associated protein (YAP) pathway. The expression and location of Amot was examined in breast cancer tissues and cell lines using immunohistochemistry, real-time polymerase chain reaction analysis (RT-PCR), western blotting and immunofluorescence. ANOVA, Student's t-test, Wilcoxon and Chi-square tests were utilized to determine the association of Amot expression with clinically relevant parameters. Stable Amot knockdown MCF-7 cells (MCF-7 Amot KD) were generated to investigate the impact of Amot downregulation on the growth and invasion of MCF-7 cells in vitro. Western blotting was applied to detect the expression of the Hippo-YAP pathway protein in the MCF-7 cells. It was observed that Amot was highly expressed in breast cancer tissues, but weakly expressed in adjacent non-cancerous tissues. Additionally, the expression level of Amot was correlated with that of Ki-67. In MCF-7 cells, Amot downregulation resulted in a significant decrease of cell proliferation and invasiveness. Following Amot knockdown in MCF-7 cells, the expression of YAP, YAP/TAZ and LATS1 was decreased. In particular, the expression of YAP was markedly reduced in the nucleoprotein. The results suggested that Amot was highly expressed in breast cancer tissues and was important in the promotion of breast cancer cell proliferation and invasion. In addition, there was a more intimate connection between Amot and Hippo-YAP pathway.

Self-assembly of nanowires into micro-scale patterns, especially in a controlled manner, has received increasing research interest because of the wide variety of potential applications, including micro-optics and electronic devices, as well as nanomaterials-based energy conversion systems. In this contribution, a novel laser-assisted solution spreading method was developed to fabricate and self-assemble alumina nanowires (ANWs) into large-scale 3-dimensional (3D) micro-patterned surfaces in one step. Here, sodium hydroxide (NaOH) solution played a dual role, both chemically etching the anodic aluminum oxide template (AAO) into ANWs and self-assembling the as-obtained ANWs into micro-patterns under capillary force. It is notable that the micro-scale patterns can be artificially controlled by introducing laser points before solution spreading on the AAO template, and thus the laser-etched area will act as the fixation point during the ANW assembly process. Moreover, the as-prepared micro-patterned ANW film exhibits typical micro-/nano-hierarchical surface topology and shows superhydrophilicity. The film can be transformed into a superhydrophobic surface by chemical modification with 1H,1H,2H,2H-perfluorodecyltriethoxysilane (FAS). Here, by taking advantage of wetting and dewetting processes of a solution on an AAO template, we propose a facile method that enables the fabrication of 3D micro-patterned ANW surfaces, which have superwetting properties. We envisage that this method could shed new light on the fabrication of functional micro-patterned devices where a one-dimensional nano-material and solution phase are involved.

C-type cytochromes located on the outer membrane (OMCs) of genus Shewanella act as the main redox-active species to mediate extracellular electron transfer (EET) from the inside of the outer membrane to the external environment: the central challenge that must be met for successful EET. The redox states of OMCs play a crucial role in dictating the rate and extent of EET. Here, we report that the surface wettability of the electrodes strongly influences the EET activity of living organisms of Shewanella loihica PV-4 at a fixed external potential: the EET activity on a hydrophilic electrode is more than five times higher than that on a hydrophobic one. We propose that the redox state of OMCs varies significantly at electrodes with different wettability, resulting in different EET activities.

PECTATE LYASE-LIKE10 (PLL10) was previously identified as one of the differentially expressed genes both in microspores during the late pollen developmental stages and in pistils during the fertilization process in Chinese cabbage (Brassica campestris ssp. chinensis). Here, antisense-RNA was used to study the functions of BcPLL10 in Chinese cabbage. Abnormal pollen was identified in the transgenic lines (bcpll10-4, -5, and -6). In fertilization experiments, fewer seeds were harvested when the antisense-RNA lines were used as pollen donor. In vivo and in vitro pollen germination assays less germinated pollen tubes were observed in bcpll10 lines. Scanning electron microscopy observation verified that the tryphine materials were over accumulated around the pollen surface and sticked them together in bcpll10. Moreover, transmission electron microscopy observation revealed that the internal endintine was overdeveloped and predominantly occupied the intine, and disturbed the normal proportional distribution of the two layers in the non-germinal furrow region; and no obvious demarcation existed between them in the germinal furrow region in the bcpll10 pollen. Collectively, this study presented a novel PLL gene that played an important role during the pollen wall development in B. campestris, which may also possess potential importance for male sterility usage in agriculture.

Splenic metastasis of ovarian cancer appears to be more common in serous cystadenocarcinomas. Splenic metastasis usually occurs postoperatively and simultaneously with dissemination to the greater omentum and pelvic cavity. Compared with other ovarian cancer subtypes, ovarian clear cell adenocarcinoma (OCCA) is rare, accounting for

The aim of this study was to explore the clinical characteristics of and treatment strategies for interstitial pneumonia induced by gefitinib in patients with advanced non-small cell lung cancer (NSCLC). The detailed clinical data of one patient with NSCLC and gefitinib-induced interstitial pneumonia were compiled and a review of relevant previous studies was performed. Based on this case report and the review, the clinical characteristics, mechanisms and treatment strategies of this rare disease were analyzed. The analyses showed that older, male patients with a long smoking history, high smoking index and adenocarcinoma (particularly bronchoalveolar carcinoma) were more likely to suffer from interstitial pneumonia while taking gefitinib. The onset time of interstitial pneumonia was 1–2 months subsequent to gefitinib administration. The clinical manifestations included chest tightness, shortness of breath, progressive dyspnea, severe hypoxemia and respiratory failure. Diffuse infiltration and alveolar interstitial shadows were observed on the chest tomography scan. In such circumstances, a timely judgment is required, in addition to the withdrawal of gefitinib treatment and the administration of high-dose glucocorticoids, as well as oxygen inhalation and anti-infective therapies, in order to relieve the symptoms. In conclusion, following the onset of gefitinib-induced interstitial pneumonia, the discontinuation of gefitinib is likely to alleviate the suffering of the majority of patients. Early interstitial pneumonia is not an absolute index for the permanent discontinuation of gefitinib treatment. It is necessary to comprehensively consider the benefits and hazards of gefitinib for the patients.

In the present study, a polygalacturonase (PG) gene was isolated from Brassica campestris L. ssp. chinensis Makino (syn. Brassica rapa ssp. chinensis). Sequence analysis indicated that it contained three classical conserved domains (I, II, and IV) of PG proteins. Phylogenetic analysis demonstrated that it was clustered into the clade E of PG family with several other PGs from different plant species. The results obtained from quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and in situ hybridization showed that the gene, B. campestris Male Fertility 24 (BcMF24), was specifically expressed in the microspores, in the partly degraded tapetum at binucleate stage and in the pollen grains at mature pollen stage. Bioinformatics analysis of a 1,489-bp sequence in the BcMF24 5′-upstream region illustrated that the said gene contained several typical cis-acting elements and pollen-specific elements. Transient expression analysis revealed that BcMF24 can drive the green fluorescent protein (GFP) expression in onion epidermal cells. β-glucuronidase (GUS) assay of the BcMF24 promoter–GUS constructs in transgenic Arabidopsis thaliana showed that it could specifically drive gene expression in the anther of flower buds at binucleate and mature pollen stages. To the best of our knowledge, the current study is the first demonstration of a PG gene, which belongs to clade E that is involved in pollen development.

Plant β-galactosidases (EC 3.2.1.23; BGAL) are associated with cell wall biogenesis and modification. β-Galactosidase isozymes are encoded by the BGAL multigene family in plants. In this study, 16 BcBGAL genes from the Brassica campestris ssp. chinensis (syn. Brassica rapa ssp. chinensis) genome were investigated and compared with those from Arabidopsis thaliana. A comprehensive genome-wide analysis of BcBGAL genes in B. campestris is presented, including their phylogeny, genome localization, gene structure, and expression patterns. Phylogenetic analysis of 16 BcBGAL and 17 AtBGAL proteins from Arabidopsis indicate that the BcBGALs clustered into four major groups, such that genes within the same group had similar exon–intron structures. The 5′-upstream region of BcBGAL genes contained a group of putative cis-acting elements related to stress and plant hormone response. Expression analysis of BcBGAL genes using quantitative reverse transcription–polymerase chain reaction demonstrates that all BcBGAL transcripts could be detected in at least one type of tissue. Some of these transcripts exhibited tissue-specific expression patterns. Our data provided insights on the evolution of the BGAL gene family and a useful reference for further functional analysis in B. campestris.

PECTATE LYASE-LIKE10 (PLL10) was previously identified as one of the differentially expressed genes both in microspores during the late pollen developmental stages and in pistils during the fertilization process in Chinese cabbage (Brassica campestris ssp. chinensis). Here, antisense-RNA was used to study the functions of BcPLL10 in Chinese cabbage. Abnormal pollen was identified in the transgenic lines (bcpll10-4, -5, and -6). In fertilization experiments, fewer seeds were harvested when the antisense-RNA lines were used as pollen donor. In vivo and in vitro pollen germination assays less germinated pollen tubes were observed in bcpll10 lines. Scanning electron microscopy observation verified that the tryphine materials were over accumulated around the pollen surface and sticked them together in bcpll10. Moreover, transmission electron microscopy observation revealed that the internal endintine was overdeveloped and predominantly occupied the intine, and disturbed the normal proportional distribution of the two layers in the non-germinal furrow region; and no obvious demarcation existed between them in the germinal furrow region in the bcpll10 pollen. Collectively, this study presented a novel PLL gene that played an important role during the pollen wall development in B. campestris, which may also possess potential importance for male sterility usage in agriculture.

Polygalacturonase (PG, EC3.2.1.15), one of the hydrolytic enzymes associated with the modification of pectin network in plant cell wall, has an important role in various cell-separation processes that are essential for plant development. PGs are encoded by a large gene family in plants. However, information on this gene family in plant development remains limited. In the present study, 53 and 62 putative members of the PG gene family in cucumber and watermelon genomes, respectively, were identified by genome-wide search to explore the composition, structure, and evolution of the PG family in Cucurbitaceae crops. The results showed that tandem duplication could be an important factor that contributes to the expansion of the PG genes in the two crops. The phylogenetic and evolutionary analyses suggested that PGs could be classified into seven clades, and that the exon/intron structures and intron phases were conserved within but divergent between clades. At least 24 ancestral PGs were detected in the common ancestor of Arabidopsis and Cucumis sativus. Expression profile analysis by quantitative real-time polymerase chain reaction demonstrated that most CsPGs exhibit specific or high expression pattern in one of the organs/tissues. The 16 CsPGs associated with fruit development could be divided into three subsets based on their specific expression patterns and the cis-elements of fruit-specific, endosperm/seed-specific, and ethylene-responsive exhibited in their promoter regions. Our comparative analysis provided some basic information on the PG gene family, which would be valuable for further functional analysis of the PG genes during plant development.

Background microRNAs (miRNAs) are endogenous, noncoding, small RNAs that have essential regulatory functions in plant growth, development, and stress response processes. However, limited information is available about their functions in sexual reproduction of flowering plants. Pollen development is an important process in the life cycle of a flowering plant and is a major factor that affects the yield and quality of crop seeds. Results This study aims to identify miRNAs involved in pollen development. Two independent small RNA libraries were constructed from the flower buds of the male sterile line (Bcajh97-01A) and male fertile line (Bcajh97-01B) of Brassica campestris ssp. chinensis. The libraries were subjected to high-throughput sequencing by using the Illumina Solexa system. Eight novel miRNAs on the other arm of known pre-miRNAs, 54 new conserved miRNAs, and 8 novel miRNA members were identified. Twenty-five pairs of novel miRNA/miRNA* were found. Among all the identified miRNAs, 18 differentially expressed miRNAs with over two-fold change between flower buds of male sterile line (Bcajh97-01A) and male fertile line (Bcajh97-01B) were identified. qRT-PCR analysis revealed that most of the differentially expressed miRNAs were preferentially expressed in flower buds of the male fertile line (Bcajh97-01B). Degradome analysis showed that a total of 15 genes were predicted to be the targets of seven miRNAs. Conclusions Our findings provide an overview of potential miRNAs involved in pollen development and interactions between miRNAs and their corresponding targets, which may provide important clues on the function of miRNAs in pollen development.

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Here, we developed a novel hybrid bio-organic interface with matchable nano-scale topography between a polypyrrole nanowire array (PPy-NA) and the bacterium Shewanella, which enabled a remarkably increased extracellular electron transfer (EET) current from genus Shewanella over a rather long period. PPy-NA thus exhibited outstanding performance in mediating bacterial EET, which was superior to normal electrodes such as carbon plates, Au and tin-doped In₂O₃. It was proposed that the combined effect of the inherent electrochemical nature of PPy and the porous structured bacterial network that was generated on the PPy-NA enabled long-term stability, while the high efficiency was attributed to the enhanced electron transfer rate between PPy-NA and microbes caused by the enhanced local topological interactions.