AeWRKY32 from okra regulates anthocyanin accumulation and cold tolerance in Arabidopsis.

Okra (Abelmoschus esculentus L.) is a tropical crop species, and its growth and development are severely affected by cold stress. Recent studies have identified a potential association between WRKY transcription factors and the cold response mechanism of crops. In this study, the AeWRKY32 transcription factor that encodes 482 amino acids was amplified from A. esculentus, and its expression level was found to be the highest in the okra flower. AeWRKY32 localized to the nucleus and displayed transcriptional activation capability. Under normal conditions, overexpression of AeWRKY32 induced anthocyanin accumulation, with higher expression levels of AtCHS1, AtCHI4, AtF3H1, and AtDFR2 in transgenic Arabidopsis. Under cold stress, anthocyanin levels were further elevated in transgenic Arabidopsis plants. At the same time, AeWRKY32 overexpression promoted ABA biosynthesis, inhibited H2O2 and O2- generation, induced stomatal closure, reduced electrolyte leakage, and thus improved the cold resistance of transgenic Arabidopsis. Furthermore, under cold stress, the expression profiles of AtCOR413, AtCOR15B, AtCBF1, and AtCBF2 were upregulated in transgenic Arabidopsis. Overall, our study provides evidence that AeWRKY32 serves as a crucial regulator in both anthocyanin accumulation and cold tolerance of transgenic Arabidopsis. Our findings could provide insights into the molecular mechanism linking AeWRKYs to plant cold tolerance.

AgMYB5, an MYB transcription factor from celery, enhanced ß-carotene synthesis and promoted drought tolerance in transgenic Arabidopsis.

BACKGROUND: Water shortage caused by global warming seriously affects the yield and quality of vegetable crops. ß-carotene, the lipid-soluble natural product with important pharmacological value, is abundant in celery. Transcription factor MYB family extensively disperses in plants and plays regulatory roles in carotenoid metabolism and water scarcity response. RESULTS: Here, the AgMYB5 gene encoding 196 amino acids was amplified from celery cv. 'Jinnanshiqin'. In celery, the expression of AgMYB5 exhibited transactivation activity, tissue specificity, and drought-condition responsiveness. Further analysis proved that ectopic expression of AgMYB5 increased ß-carotene content and promoted drought tolerance in transgenic Arabidopsis thaliana. Moreover, AgMYB5 expression promoted ß-carotene biosynthesis by triggering the expression of AtCRTISO and AtLCYB, which in turn increased antioxidant enzyme activities, and led to the decreased contents of H2O2 and MDA, and the inhibition of O2- generation. Meanwhile, ß-carotene accumulation promoted endogenous ABA biosynthesis of transgenic Arabidopsis, which resulted in ABA-induced stomatal closing and delayed water loss. In addition, ectopic expression of AgMYB5 increased expression levels of AtERD1, AtP5CS1, AtRD22, and AtRD29. CONCLUSIONS: The findings indicated that AgMYB5 up-regulated ß-carotene biosynthesis and drought tolerance of Arabidopsis.

TREM2 as a Prognostic Biomarker for Osteosarcoma Microenvironment Remodeling.

The tumor microenvironment (TME) acts as a crucial role in the occurrence and development of osteosarcoma (OS). Despite this, the mechanism controlling the components of immunity and stroma in the tumor microenvironment remains a mystery. To conduct this study, we download and collate transcriptome data from the TARGET database, whose full name is Therapeutically Applicable Research to Generate Effective Treatments, as well as available clinical information of OS. The CIBERSORT and ESTIMATE methodology are used to acquire the proportions of components of immunity and stroma and tumor-infiltrating immune cells (TICs). Protein-protein interaction (PPI) networks and Cox regression analysis are used to select differentially expressed genes (DEGs). A prognostic biomarker is determined by intersecting univariate COX and PPI results, which lead to the finding of Triggering receptor expressed on myeloid cells-2 (TREM2). Based on the next analysis, TREM2 expression is positively correlated with OS survival time. Immune function-related genes have enrichment in the group with high expression of TREM2, according to gene set enrichment analysis (GSEA). The percentage of TICs by CIBERSORT methodology revealed that the expression of TREM2 is positively associated with follicular helper T cells, CD8-positive T cells, and M2 macrophages and negatively correlated with plasma cells, M0 macrophages, and naive CD4-positive T cells. All results suggest a possible integral role of TREM2 in the immune-related events of TME. Therefore, TREM2 may be a potential indicator of remodeling of TME in osteosarcoma, which is useful and helpful in predicting the clinical prognostic outcome of OS patients and provide a unique perspective for immunotherapy for OS.

A Secure and Traceable Vehicles and Parts System Based on Blockchain and Smart Contract.

As society advances, so does the total number of vehicles on the road, creating a massive consumer market for automobiles. According to statistics, a major portion of today's traffic difficulties are caused by accidents caused by subpar cars and auto parts. As a result, each country has, over time, enacted equivalent rules and regulations to prevent such tragedies. However, in the face of profit, some people are desperate enough to employ illegal parts and illegally modified cars, and auto fraud is rampant. As a result, we employ the blockchain of the symmetrical Blockchain's digital ledger and smart contract technology to build a decentralized supply chain system that can identify specific parts. In this study, we design and discuss the proposed system framework by user functions and the flow of parts based on blockchain, and we discuss communication protocols that use the symmetry and asymmetry cryptography, algorithms, properties, and security of the mechanism while providing related analysis and comparing the properties and costs of the system with other studies. Overall, the proposed method has the potential to successfully address the issue of automobile fraud.

Comparative transcriptome analysis provides insight into nitric oxide suppressing lignin accumulation of postharvest okra (Abelmoschus esculentus L.) during cold storage.

In plants, NO has been proved the function of improving abiotic stress resistance. However, the role of NO in the lignin metabolism of okra under cold stress has not been clarified. Here, histochemical staining and lignin content analysis showed that cold stress promoted the lignin accumulation of cold stored okra pods, and NO inhibited the lignin accumulation and delayed lignification process. To better understand the roles of NO in okra cold stress resistance mechanism, the full-length transcriptome data of 'Hokkaido' was analyzed. The SNP-treated okra transcriptome and cPTIO-treated okra transcriptome were obtained. A total of 41957 unigenes were screened out from three groups at 10 d, among which, 33, 78 and 18 DEGs were found in ddH2O-treat, SNP-treat and cPTIO-treat group, respectively. Transcriptomic data suggested that the genes involved in lignin biosynthesis showed downregulation under SNP treatment. Transcriptomic data and enzyme activity showed that exogenous NO significantly promoted the biosynthesis of endogenous NO by enhancing NOS activity. Transcriptomic data and plant hormone data showed that NO played an important role in the process of inhibiting the ethylene and ABA synthesis mechanism of okra and thereby reducing the endogenous ethylene and ABA content under chilling stress. Relevant physiological data showed that NO helped to the protection of ROS scavenging system and removed the MDA and H2O2 induced by cold stress. These results provided a reference for studying the molecular mechanism of nitric oxide delaying the lignification of okra, and also provided a theoretical basis for postharvest storage of vegetables.

TMBIM1 is an inhibitor of adipogenesis and its depletion promotes adipocyte hyperplasia and improves obesity-related metabolic disease.

Obesity is characterized by the excessive accumulation of the white adipose tissue (WAT), but healthy expansion of WAT via adipocyte hyperplasia can offset the negative metabolic effects of obesity. Thus, identification of novel adipogenesis regulators that promote hyperplasia may lead to effective therapies for obesity-induced metabolic disorders. Using transcriptomic approaches, we identified transmembrane BAX inhibitor motif-containing 1 (TMBIM1) as an inhibitor of adipogenesis. Gain or loss of function of TMBIM1 in preadipocytes inhibited or promoted adipogenesis, respectively. In vivo, in response to caloric excess, adipocyte precursor (AP)-specific Tmbim1 knockout (KO) mice displayed WAT hyperplasia and improved systemic metabolic health, while overexpression of Tmbim1 in transgenic mice showed the opposite effects. Moreover, mature adipocyte-specific Tmbim1 KO did not affect WAT cellularity or nutrient homeostasis. Mechanistically, TMBIM1 binds to and promotes the autoubiquitination and degradation of NEDD4, which is an E3 ligase that stabilizes PPARγ. Our data show that TMBIM1 is a potent repressor of adipogenesis and a potential therapeutic target for obesity-related metabolic disease.

Half-Brain Delineation for Prediction of Radiation-Induced Temporal Lobe Injury in Nasopharyngeal Carcinoma Receiving Intensity-Modulated Radiotherapy.

PURPOSE: To investigate the role of half-brain delineation in the prediction of radiation-induced temporal lobe injury (TLI) in nasopharyngeal carcinoma (NPC) receiving intensity-modulated radiotherapy (IMRT). METHODS AND MATERIALS: A total of 220 NPC cases treated with IMRT and concurrent platinum-based chemotherapy were retrospectively analyzed. Dosimetric parameters of temporal lobes, half-brains, and brains included maximum dose (Dmax), doses covering certain volume (DV) from 0.03 to 20 cc and absolute volumes receiving specific dose (VD) from 40 to 80 Gy. Inter-structure variability was assessed by coefficients of variation (CV) and paired samples t-tests. Receiver operating characteristic curve (ROC) and Youden index were used for screening dosimetric parameters to predict TLI. Dose/volume response curve was calculated using the logistic dose/volume response model. RESULTS: CVs of brains, left/right half-brains, and left/right temporal lobes were 9.72%, 9.96%, 9.77%, 27.85%, and 28.34%, respectively. Each DV in temporal lobe was significantly smaller than that in half-brain (P < 0.001), and the reduction ranged from 3.10% to 45.98%. The area under the curve (AUC) of DV and VD showed an "increase-maximum-decline" behavior with a peak as the volume or dose increased. The maximal AUCs of DVs in brain, half-brain and temporal lobe were 0.808 (D2cc), 0.828 (D1.2cc) and 0.806 (D0.6cc), respectively, and the maximal AUCs of VDs were 0.818 (D75Gy), 0.834 (V72Gy) and 0.814 (V70Gy), respectively. The cutoffs of V70Gy (0.86 cc), V71Gy (0.72 cc), V72Gy (0.60 cc), and V73Gy (0.45 cc) in half-brain had better Youden index. TD5/5 and TD50/5 of D1.2cc were 58.7 and 80.0 Gy, respectively. The probability of TLI was higher than >13% when V72Gy>0 cc, and equal to 50% when V72Gy = 7.66 cc. CONCLUSION: Half-brain delineation is a convenient and stable method which could reduce contouring variation and could be used in NPC patients. D1.2cc and V72Gy of half-brain are feasible for TLI prediction model. The dose below 70 Gy may be relatively safe for half-brain. The cutoff points of V70-73Gy could be considered when the high dose is inevitable.

Landscape pattern evolution along terrain gradient in Fuzhou City, Fujian Province, China.

Terrain is an important factor in land landscape pattern change. To reveal the spatial and temporal characteristics and variation of landscape pattern along a terrain gradient, we used three remote sensing images and digital elevation model (DEM) data of Fuzhou in 1995, 2005 and 2015 to investigate the topographic gradient effect of landscape pattern and the cause of formation based on terrain index, land use distribution index, geo-informatics map analysis and landscape index. The results showed that forestland was mainly distributed in middle-low, middle-high and higher level terrain, while farmland, water body, construction land and unused lands were mainly distributed in the low rank terrain. From 1995 to 2015, the area of forestland, farmland, and unused land in Fuzhou decreased, while that of construction land and water body increased. The change of landscape type was stable, which was mainly distributed in middle-low, middle-high, and high grade terrain gradient region. In addition, landscape pattern changes were obvious across the terrain gra-dient. The landscape type of low terrain area mainly shifted to the construction land, but farmland and forestland had an alternation change in the middle-low and middle-high terrain area. The characteristics of landscape pattern fragmentation, landscape heterogeneity and landscape diversity in the study area had been increasing year by year, but they had decreased with the elevation of terrain.

[Influence of ENSO Events on the Hydrogen (δ2H) and Oxygen (δ18O) Isotopic Values of Precipitation in Shanghai].

An analysis of the effect of ENSO events with different strengths on the isotopic composition of precipitation is conducted based on test data for 206 precipitation samples collected from January 2012 to February 2017 in Shanghai coupled with the archives for Nanjing, Wuhan, Fuzhou, and Hong Kong from 1961 to 2012 from the Global Network of Isotopes in Precipitation (GNIP) database. During the research periods, the δD and δ18O values in precipitation are lower in summer and autumn but higher in winter and spring. The slope and intercept of the atmospheric precipitation lines during El Niño events are larger than during other times, while anti-temperature, precipitation amount, and vapor pressure effects are more significant than during La Niña events. The δ18O and deuterium excess values (value of d) of rainfall in Shanghai during El Niño and La Niña events of varied strengths have an obvious negative correlation with the oceanic Niño index (ONI), sea surface temperature anomaly (SSTA), and the extreme and cumulative values of ONI. Moreover, ENSO events are closely intertwined with the correlation between δ18O isotopic value in precipitation, ONI, and SSTA.

[Characteristics of Stable Isotope in Precipitation and Its Relationship with ENSO in Shanghai].

A total of 70 precipitation samples were collected from August 2014 to August 2015 in Shanghai and the oxygen and hydrogen isotope of precipitation were analyzed. The relationship between the δ18O and deuterium excess of precipitation and ENSO events was addressed. The results showed that δD and δ18O of precipitation in Shanghai were higher in winter and spring than in summer and fall. The slope of meteoric water line with these data was a little smaller than the global meteoric water line. The main reason could be the unbalanced secondary evaporation in the process of precipitation. The correlation between the δ18O in precipitation and temperature and precipitation varied in different time scales. On a seasonal scale, this work revealed that there was no significant temperature effect in winter, but significant precipitation effect was found in the annual scales, which was obviously affected by the atmospheric circulation. δ18O values and deuterium excess in precipitation recorded the La Niña and El Niño conversion process. During the La Niña event, the δ18O and d values were more negative than those in the El Niño event.

Distribution of As, Cd, and Pb in seafood in Southern China and their oral bioavailability in mice.

The distribution of the toxic elements As, Cd, and Pb in nine different types of seafood from Shenzhen, China, was investigated by using inductively coupled plasma mass spectrometry (ICP-MS). The results revealed that the concentrations of arsenic (As) in fish (Lutjanus erythropterus, Paralichthys olivaceus) and in bivalve (Meretrix meretrix) and cadmium (Cd) in scallop (Argopecten irradians) exceed the limits established by food safety regulations in China and EU (European Union). Furthermore, the bioavailability of As, Cd, and lead (Pb) in mice after 20-day oral ingestion of Crassostrea rivularis was investigated, and the total rate of absorption of toxic elements in samples from the liver and kidney tissues and blood was determined. The results of this in vivo trial indicated that the oral bioavailability of As, Cd, and Pb was approximately 0.33, 0.45, and 0.74%, respectively.

Treatment responses of procaterol and CD38 inhibitors in an ozone-induced airway hyperresponsiveness mice model.

Airway hyperresponsiveness (AHR) and airway inflammation are key pathophysiological features of many respiratory diseases, such as asthma and chronic obstructive pulmonary disease (COPD). To evaluate the treatment responses of procaterol and CD38 inhibitors in an ozone-induced AHR mice model, we hypothesized that procaterol and two synthetic CD38 inhibitors (Compounds T and H) might have therapeutic effects on the ozone-induced AHR mice model, and the nuclear factor-kappaB (NF-κB) pathway and the CD38 enzymatic activity might be involved in the mechanisms. With the exception of the Control group, ozone exposure was used to establish an AHR model. Male Kunming mice in the Procaterol and CD38 inhibitors groups were treated with an emulsifier of procaterol hydrochloride, Compound T or H. Results indicated that (1) no drug showed severe toxicity in this study; (2) ozone exposure induced airway inflammation and AHR; (3) intragastric treatment with procaterol and Compound T achieved potent therapeutic effects, but Compound H did not show any therapeutic effect; (4) the NF-κB pathway was involved in both the pathogenic mechanisms of ozone and therapeutic mechanisms of procaterol and Compound T; (5) however, the in vivo effect of Compound T was not caused by its inhibitory activity on CD38. Taken together, procaterol and Compound T are potentially good drugs to treat asthma and COPD complicated with ozone exposure.