INTRODUCTION:Homeobox genes are highly conserved and play critical roles in brain development. Recently, we have found that mammals have an additional fragment of approximately 20 amino acids in Emx1 and a poly-(AL)6-7 in Emx2, compared to other amniotes. It has been shown that Emx1 and Emx2 have synergistic actions in the brain development. These reports raise an interesting issue whether the differences of Emx1 and Emx2 between mammals and non-mammals are concerned with the organization and evolution of amniote pallium. METHODS:Lentiviruses expressing mouse Emx1 and Emx2 (mEmx1/2) with additional fragments were injected into the ventricle of the chick telencephalon at embryonic day 3 to study the effects of mEmx1/2 on the development of chick pallium, whereas injections of lentiviruses containing chick Emx1 and Emx2 (cEmx1/2), no targeted gene insert or saline were as controls. The expressions of reelin, vimentin, GABA and MAP2, neurogenesis patterns for calbindin (CB) and parvalbumin (PV) neurons and the sizes of anterior commissure (AC) were then studied by immuohistochemical staining, and open-field tests were performed to assess locomotor activities and curious or exploratory behaviors of the chicks. RESULTS:Following the injections of lentiviruses expressing mEmx1/2, the expressions of reelin, vimentin, GABA, and MAP2 increased in most parts of Wulst (W) and mesopallium (M), but not most of nidopallium (N). Neurogenesis patterns for CB and PV neurons changed toward mammalian inside-out one, and the sizes of AC staining for neurofilament were significantly larger. In addition, post-hatchling chicks showed higher rates of passive avoidance after training, but no significant differences in the total distance traveled and the percentage of time spent in the central rectangle, compared to those in the control groups. CONCLUSION:The present study indicated that mEmx1/2 had effects on the development of chick pallium, suggesting that they are probably involved in the organization and evolution of amniote pallium.

Objective: To explore the effects of Brazilin on the Human periodontal ligament stem cells (hPDLSCs) mediating periodontal tissue regeneration in periodontitis and the Yes-associated protein (YAP) pathway for periodontitis. Methods: The anti-inflammatory effects and osteogenic effects of Brazilin in the inflammatory microenvironment were mainly evaluated by polymerase chain reaction (PCR), western blot (WB), staining, and the blocking and deletion of the YAP pathway in vitro by verteporfin (VP) and shRNA to evaluated the effect on the YAP pathway. A rat periodontitis model was established and the therapeutic effects were evaluated by Micro-CT, hematoxylin and eosin (HE) staining, Masson staining, and immunohistochemistry. Results: Brazilin effectively down-regulated the expression of inflammatory factors, showed deeper staining, and showed up-regulation of related osteogenic factors, and inhibition of the YAP pathway is followed by inhibition of the osteogenic effects of Brazilin. Brazilin was effective in reducing inflammation and inhibiting alveolar bone loss in the rat periodontitis model. Conclusion: Brazilin can promote periodontal tissue regeneration in periodontitis through the YAP pathway, providing new ideas for the treatment of clinical periodontitis.

There is obvious sexual dimorphism in the song control system of songbirds. In the higher vocal center (HVC), cell proliferation and neuronal differentiation contribute to the net addition of neurons. However, the mechanism underlying these changes is unclear. Given that Wnt, Bmp, and Notch pathways are involved in cell proliferation and neuronal differentiation, no reports are available to study the role of the three pathways in the song control system. To address the issue, we studied cell proliferation in the ventricle zone overlying the developing HVC and neural differentiation within the HVC of Bengalese finches (Lonchura striata) at posthatching day 15 when HVC progenitor cells are generated on a large scale and differentiate into neurons, after Wnt and Bmp pathways were activated by using a pharmacological agonist (LiCl) or Bmp4, respectively, and the Notch pathway was inhibited by an inhibitor (N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester), DAPT). The results indicated that both cell proliferation and neural differentiation toward HVC neurons increased significantly after activation of the Wnt signaling pathway or inhibition of the Notch signaling pathway. Although cell proliferation was increased, neural differentiation was inhibited after treatment with Bmp4. There was obvious synergetic enhancement in the number of proliferating cells after the coregulation of two or three signaling pathways. In addition, synergetic enhancement was also found in the Wnt and Notch pathways in neural differentiation toward neurons within HVC. These results suggest that the three signaling pathways are involved in cell proliferation and neural differentiation of HVC.

Objective:To investigate the construction of ostomy nurse-led clinics in tertiary hospitals in China.Methods:Using the convenient method, managers of ostomy nurse-led clinics in tertiary hospitals in 31 provinces, autonomous regions and municipalities in China in August 2020 were selected as the survey objects. Questionnaire for Survey on Status Quo of Ostomy nurse-led clinics in ClassⅢ Grade A hospital in China was used to investigate. A total of 367 questionnaires were recovered in this survey, 313 valid questionnaires were recovered, and the effective recovery rate was 85.29%.Results:A total of 313 tertiary hospitals have set up ostomy nurse-led clinics. Among the ostomy nurse-led clinics, 86.58% (271/313) of the ostomy specialist nursing outpatient visits were independent visits by nurses. The opening period of stoma clinic was (6.27±4.08) years and the weekly opening time was (5.30±3.93) half-days. The average monthly outpatient number was 100, with a minimum of 4 and a maximum of 3 000. In terms of job management, 50.48% (158/313) of the stoma specialist nursing outpatient jobs were part-time, managers were mostly undergraduates (74.44%, 233/313) , the professional title of managers was mostly nurse in charge (48.24%, 151/313) , 38.98% (122/313) of hospitals had promotion system and 36.74% (115/313) of hospitals had performance management system. There were large differences in environmental layout, facilities and equipment, goods management and specialist management and there was no unified standard.Conclusions:There are great differences in the construction of ostomy nurse-led clinics in tertiary hospitals in China, which need to be further standardized and unified. It is suggested that relevant departments should introduce relevant policies to encourage the establishment of full-time nurse posts in ostomy nurse-led clinics, guarantee the training, promotion and performance reward of visiting nurses and fully encourage the subjective initiative of ostomy specialist nurses to promote their career development.

Ethylene response factor (ERF) Group VII members generally function in regulating plant growth and development, abiotic stress responses, and plant immunity in Arabidopsis; however, the details of the regulatory mechanism by which Group VII ERFs mediate plant immune responses remain elusive. Here, we characterized one such member, ERF72, as a positive regulator that mediates resistance to the necrotrophic pathogen Botrytis cinerea. Compared with the wild-type (WT), the erf72 mutant showed lower camalexin concentration and was more susceptible to B. cinerea, while complementation of ERF72 in erf72 rescued the susceptibility phenotype. Moreover, overexpression of ERF72 in the WT promoted camalexin biosynthesis and increased resistance to B. cinerea. We identified the camalexin-biosynthesis genes PAD3 and CYP71A13 and the transcription factor WRKY33 as target genes of ERF72. We also determined that MPK3 and MPK6 phosphorylated ERF72 at Ser151 and improved its transactivation activity, resulting in increased camalexin concentration and increased resistance to B. cinerea. Thus, ERF72 acts in plant immunity to coordinate camalexin biosynthesis both directly by regulating the expression of biosynthetic genes and indirectly by targeting WRKK33.

Objective:To compile the post quality standard scale for wound, ostomy and continence nurses in China and to test the reliability and validity of the scale.Methods:The first draft of the scale was formed through literature review, semi-structured interview and Delphi expert letter consultation. Using the purposive sampling method, a total of 796 wound, ostomy and continence nurses from 31 provinces, municipalities and autonomous regions of China were selected from April to May 2019 to test the reliability and validity of the scale. A total of 796 questionnaires were sent out and 693 were effectively received, with effective recovery of 87.1%.Results:The post quality standard scale for wound, ostomy and continence nurses included 3 first-level items and 77 second-level items. Three common factors were extracted from exploratory factor analysis and the cumulative variance contribution rate was 72.466%. The correlation coefficient between the first-level items was 0.709-0.898 and the correlation coefficient between the first-level item and the total scale was 0.859-0.937. The Cronbach's α coefficient of the total amount table was 0.992 and the half-fold reliability coefficient was 0.917 ( P<0.01) . Conclusions:The post quality standard scale for wound, ostomy and continence nurses has good reliability and validity, which can be used to evaluate the post quality of wound, ostomy and continence nurses.

Drought and salt are two major abiotic stresses that severely impact plant growth and development, as well as crop production. A previous study showed that OsOSCA1.4, one of eleven rice OSCAs (OsOSCAs), complements hyperosmolality-induced [Ca2+](cyt) increases (OICIcyt), salt stress-induced [Ca2+](cyt) increases (SICIcyt) and the associated growth phenotype in Arabidopsis osca1 (reduced hyperosmolality-induced [Ca2+](cyt) increase 1). In this study, Except for OsOSCA2.3 and OsOSCA4.1, we generated independent transgenic lines overexpressing eight other OsOSCAs in the osca1 to explore their functions in osmotic Ca2+ signalling, stomatal movement, leaf water loss, and root growth in response to hyperosmolality and salt stress. Similar to OsOSCA1.4, overexpression of OsOSCA1.1 or OsOSCA2.2 in osca1 complemented OICIcyt and SICIcyt, as well as stomatal closure and root growth in response to hyperosmolality and salt stress treatments, and drought-related leaf water loss. In addition, overexpression of OsOSCA1.2, OsOSCA1.3 or OsOSCA2.1 in osca1 restored OICIcyt and SICIcyt, whereas overexpression of OsOSCA2.5 or OsOSCA3.1 did not. Moreover, osca1 overexpressing these five OsOSCAs exhibited various abiotic stress-associated growth phenotypes. However, overexpression of OsOSCA2.4 did not have any of these effects. These results indicated that multiple members of the OsOSCA family have redundant functions in osmotic sensing and diverse roles in stress adaption.

Nitrogen discharged from urban areas greatly deteriorates the water quality of downstream surface water. In this study, sub-hourly high-frequency samples of urban runoff during six rainfall events were collected at the outfall of the stormwater network in an urban watershed in Beijing to explore nitrate export and transportation. Isotopic values of local road dust, soil, and network sediment were measured and used as the sources of nitrate to better elucidate the sources of the urban watershed. The contributions of various sources over and during three rainfall events with different grades were quantified and compared. The results showed that the contribution of sources changed dramatically over and during rainfall events. Along with the increase in the total rainfall amount and the going on of rainfall events, the wash-off effect in the atmosphere and on land surfaces played a more important role in nitrate output. Atmospheric deposition was the dominant contributor of nitrate in heavy and storm events (mean 59.3% and 64.8%, respectively). Network sediment contributed large proportions of nitrate in moderate and heavy events (mean 35.6% and 15.9%, respectively). The contribution of soil increased substantially in the storm event (mean 26.1%). Road dust and network sediment contributed greatly in the early stage of the heavy event. The contribution of fertilizer in heavy events was mainly because of the wash-off of road dust. The changing pattern of sources from atmospheric deposition to inorganic N fertilizer existed during the process of the storm event. The contribution of NO3- fertilizer from soil surfaces increased substantially in the later stage of the storm event. These results provide valuable references for urban nutrient management and mitigation measures implementation.

Genetically Encoded Calcium Indicators (GECIs) are powerful molecular tools for monitoring calcium (Ca2+) signaling in the cytosol and organellar compartments. However, currently available ratiometric indicators that allow measurements of resting Ca2+ levels have limitations in long-term Ca2+ imaging. They either are ultraviolet (UV)-excited ones with strong photo-toxicity, or have poor performance. To overcome this hurdle, we developed a set of visible light excited ratiometric-GECIs (VR-GECIs) based on existing mono-colored GECIs. With performance comparable to their corresponding mono-color prototypes, this set of VR-GECIs enables long-term measurements of intra-cellular or intra-organellar Ca2+ signals. Using these VR-GECIs together with a newly developed off-line analysis tool, we achieved long-term measurements of Ca2+ homeostasis of moving or dividing cells. Our tools may find broad applications in decoding Ca2+-modulated physiological or pathological processes.

In rice (Oryza sativa), caryopses located on proximal secondary branches (CSBs) have smaller grain size and poorer grain filling than those located on apical primary branches (CPBs), greatly limiting grain yield. However, the molecular mechanism responsible for developmental differences between CPBs and CSBs remains elusive. In this transcriptome-wide expression study, we identified the gene Aspartic Protease 1 (OsAsp1), which reaches an earlier and higher transcriptional peak in CPBs than in CSBs after pollination. Disruption of OsAsp1 expression in the heterozygous T-DNA line asp1-1+/-eliminated developmental differences between CPBs and CSBs. OsAsp1 negatively regulated the transcriptional inhibitor of auxin biosynthesis, OsTAA1 transcriptional inhibition factor 1 (OsTIF1), to preserve indole-3-acetic acid (IAA) apical dominance in CPBs and CSBs. IAA also facilitated OsTIF1 translocation from the endoplasmic reticulum (ER) to the nucleus by releasing the interaction of OsTIF1 with OsAsp1 to regulate caryopses IAA levels via a feedback loop. IAA promoted transcription of OsAsp1 through MADS29 to maintain an OsAsp1 differential between CPBs and CSBs during pollination. Together, these findings provide a mechanistic explanation for the distributed auxin differential between CPBs and CSBs to regulate distinct caryopses development in different rice branches and potential targets for engineering yield improvement in crops.

Obesity and its related metabolic disorders are closely correlated with gut dysbiosis. Montmorillonite is a common medicine used to treat diarrhea. We have previously found that dietary lipid adsorbent-montmorillonite (DLA-M) has an unexpected role in preventing obesity. The aim of this study was to further investigate whether DLA-M regulates intestinal absorption and gut microbiota to prevent obesity-related metabolic disorders. Here, we show that DLA-M absorbs free fatty acids (FFA) and endotoxins in vitro and in vivo. Moreover, the combination of fluorescent tracer technique and polarized light microscopy showed that DLA-M crystals immobilized BODIPY® FL C16 and FITC-LPS, respectively, in the digestive tract in situ. HFD-fed mice treated with DLA-M showed mild changes in the composition of the gut microbiota, particularly increases in short-chain fatty acids (SCFA)-producing Blautia bacteria and decreases in endotoxin-producing Desulfovibrio bacteria, these changes were positively correlated with obesity and inflammation. Our results indicated that DLA-M immobilizes FFA and endotoxins in the digestive tract via the adsorption-excretion axis and DLA-M may potentially be used as a prebiotic to prevent intestinal dysbiosis and obesity-associated metabolic disorders in obese individuals.

Lipid‐filled mature adipocytes are important for the study of lipid metabolism and in the development of obesity, but whether they are capable of reproliferation is still controversial. Here, we monitored lipid droplet dynamics and adipocyte reproliferation in live, differentiated 3T3‐L1 cells using a phase‐contrast microscope in real time. Phase‐contrast microscopy achieves a similar visual effect in situ to that obtained using traditional dyes such as Oil Red O and BODIPY in vitro. Using this method, we captured the process that lipid droplets use for dynamic fusion in living cells. Unexpectedly, we acquired images of the moment that differentiated 3T3‐L1 cells containing lipid droplets entered mitosis. In addition, we observed some binucleated mature adipocytes. This information provides a better understanding of the adipocyte differentiation process.

Protein phosphatase 4 (PP4) was shown to participate in multiple cellular processes, including DNA damage response, cell cycle and embryo development. Recent studies demonstrated a looming role of PP4 in glucose metabolism. However, whether PP4 is involved in hepatic insulin resistance remains poorly understood. The objective of this study was to estimate the role of PP4 in tumor necrosis factor (TNF)-α-induced hepatic insulin resistance. db/db mice and TNF-α-treated C57BL/6J mice were used as hepatic insulin resistance animal models. In vitro models were established in both HepG2 cells and primary hepatocytes by TNF-α treatment. We found that increased expression and activity of PP4 occurred in the livers of db/db mice and TNF-α-induced hepatic insulin resistance both in vitro and in vivo. Actually, PP4 silencing and suppression of PP4 activity ameliorated TNF-α-induced hepatic insulin resistance, whereas over-expression of PP4 caused insulin resistance. We then further investigated the prodiabetic mechanism of PP4 in TNF-α-induced insulin resistance. We found that PP4 formed a complex with IRS-1 to promote phosphorylation of IRS-1 on serine 307 via JNK activation and reduce the expression of IRS-1. Thus, PP4 is an important regulator in inflammatory related insulin resistance.

NAC with a transmembrane (TM) motif1-like (NTL) transcription factors, containing three regions: the N-terminal NAC domain (ND), the middle regulation region (RR), and the C-terminal TM domain, belong to the tail-anchored proteins. Although these NTLs play numerous essential roles in plants, their subcellular distribution and the mechanism of translocation into the nucleus (NU) remain unclear. In this study, we found that most of the full-length NTLs were localized in the endoplasmic reticulum (ER), with the exception of NTL11 and NTL5, which were restricted to the NU. Furthermore, we found that NTL11 contains a TM domain, whereas NTL5 does not. The ND of all of the NTLs was responsible for nuclear localization in plants. After truncation of the TM domain, NTL8_NR, NTL10_NR and NTL13_NR localized in the cytoplasm (CT) and NU, and other NTL_NRs were only localized in the NU, suggesting that the RR of NTL8, NTL10 and NTL13 contains some inhibitory region to mask the nuclear localization signal sequence in the ND domain and permit their diffusion between CT and NU. Furthermore, the N-terminus of NTL11 was translocated to the NU, but the C-terminus was degraded in Arabidopsis mesophyll protoplasts. The chimeric construct of NTL11_ND with NTL10_RR and TM domain (11ND-10RT) was localized exclusively in the ER, and not in the NU. However, 10ND-11RT was found mainly in the NU. Our results indicated that the TM domain is essential for NTL targeting the ER and the N-terminal fragment, including ND and RR, is translocated into the NU after activation through proteolytic cleavage events upon stimulation by internal and external environmental signals.

MicroRNA167 (miR167), as a conserved miRNA, has been implicated in auxin signalling by regulating the expression of certain auxin response factor (ARF) genes to determine the plant developmental process. Among the 10 MIR167 genes of rice, the precursor structures derived from MIR167a, MIR167b and MIR167c produce miR167 with high efficiency. To explore the biological function of miR167 in rice, four of its predicted target genes, OsARF6, OsARF12, OsARF17 and OsARF25, were identified in vivo. Although the expression levels of miR167 and its target OsARFs did not show an obvious negative correlation, the enhanced miR167 level in transgenic rice overexpressing miR167 resulted in a substantial decrease in mRNA levels of the four OsARF genes. Moreover, the transgenic rice plants were small in stature with remarkably reduced tiller number. These results suggest that miR167 is important for the appropriate expression of at least four OsARFs, which mediate the auxin response, to contribute to the normal growth and development of rice.

The binding properties of [Ru(bpy)(2)(H2IIP)](2+) (1) {bpy = 2,2'-bipyridine, H2IIP = 2-(indole-3-yl)-imidazolo[4,5-f][1,10]phenanthroline} with calf thymus DNA (CT-DNA) and yeast tRNA have been investigated comparatively by different spectroscopic and viscosity measurements. The results suggest that the affinity of complex 1 binding with yeast tRNA is stronger than that of complex 1 binding with CT-DNA, and complex 1 is a better enantioselective binder to yeast tRNA than to CT-DNA. The toxicity of complex 1 was concentration dependent, and HL-60 cells are more sensitive to complex 1 than Hep-G2 cells; complex 1 could induce Hep-G2 cell apoptosis.