Impact of wildfire ash on skin and gut microbiomes and survival of Rana dybowskii.

Climate change and human activities escalate the frequency and intensity of wildfires, threatening amphibian habitats and survival; yet, research on these impacts remains limited. Wildfire ash alters water quality, introduces contaminants, and may disrupt microbial communities, impacting gut and skin microbiota; however, the effects on gut and skin microbiota remain unclear. Rana dybowskii were exposed to five concentrations (0 g L-1, 1.25 g L-1, 2.5 g L-1, 5 g L-1, and 10 g L-1) of aqueous extracts of wildfire ashes (AEAs) for 30 days to assess AEAs' metal content, survival, and microbiota diversity via Illumina sequencing. Our results showed that the major elements in ash were Ca > K > Mg > Al > Fe > Na > Mn, while in AEA they were K > Ca > Na > Mg > As > Al > Cu. A significant decrease in amphibian survival rates with increased AEA concentration was shown. The beta diversity analysis revealed distinct shifts in microbiota composition. Notably, bacterial genera associated with potential health risks showed increased abundance in skin microbiota, emphasising the potential for ash exposure to affect amphibian health. Functional prediction analyses revealed significant shifts in metabolic pathways related to health and disease, indicating that wildfire ash exposure may influence amphibian health through changes in microbial functions. This study highlights the urgent need for strategies to mitigate wildfire ash impacts on amphibians, as it significantly alters microbiota and affects their survival and health.

Rice straw ash and amphibian health: A deep dive into microbiota changes and potential ecological consequences.

Rice straw is burned as a result of agricultural practices and technical limitations, generating significant volumes of ash that might have environmental and ecological consequences; however, the effects on organisms have not been researched. Amphibians depend on their gut and skin microbiomes. Ash exposure may cause inflammation and changes in microbial diversity and function in frogs' skin and gut microbiota due to its chemical composition and physical presence, but the implications remain unclear. Rana dybowskii were exposed to five aqueous extracts of ashes (AEA) concentrations for 30 days to study survival, metal concentrations, and microbial diversity, analyzing the microbiota of the cutaneous and gut microbiota using Illumina sequencing. Dominant elements in ash: K > Ca > Mg > Na > Al > Fe. In AEA, K > Na > Ca > Mg > As > Cu. Increased AEA concentrations significantly reduced frog survival. Skin microbiota alpha diversity varied significantly among all treatment groups, but not gut microbiota. Skin microbiota differed significantly across treatments via Bray-Curtis and weighted UniFrac; gut microbiota was only affected by Bray-Curtis. Skin microbiota varied significantly with AEA levels in Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes, while the gut microbiota's dominant phyla, Firmicutes, Bacteroidetes, and Proteobacteria, remained consistent across all groups. Lastly, the functional prediction showed that the skin microbiota had big differences in how it worked and looked, which were linked to different health and environmental adaptation pathways. The gut microbiota, on the other hand, had smaller differences. In conclusion, AEA exposure affects R. dybowskii survival and skin microbiota diversity, indicating potential health and ecological impacts, with less effect on gut microbiota.

Tumorigenesis of basal muscle invasive bladder cancer was mediated by PTEN protein degradation resulting from SNHG1 upregulation.

BACKGROUND: Phosphatase and tensin homolog deleted on chromosome ten (PTEN) serves as a powerful tumor suppressor, and has been found to be downregulated in human bladder cancer (BC) tissues. Despite this observation, the mechanisms contributing to PTEN's downregulation have remained elusive. METHODS: We established targeted genes' knockdown or overexpressed cell lines to explore the mechanism how it drove the malignant transformation of urothelial cells or promoted anchorageindependent growth of human basal muscle invasive BC (BMIBC) cells. The mice model was used to validate the conclusion in vivo. The important findings were also extended to human studies. RESULTS: In this study, we discovered that mice exposed to N-butyl-N-(4-hydroxybu-tyl)nitrosamine (BBN), a specific bladder chemical carcinogen, exhibited primary BMIBC accompanied by a pronounced reduction in PTEN protein expression in vivo. Utilizing a lncRNA deep sequencing high-throughput platform, along with gain- and loss-of-function analyses, we identified small nucleolar RNA host gene 1 (SNHG1) as a critical lncRNA that might drive the formation of primary BMIBCs in BBN-treated mice. Cell culture results further demonstrated that BBN exposure significantly induced SNHG1 in normal human bladder urothelial cell UROtsa. Notably, the ectopic expression of SNHG1 alone was sufficient to induce malignant transformation in human urothelial cells, while SNHG1 knockdown effectively inhibited anchorage-independent growth of human BMIBCs. Our detailed investigation revealed that SNHG1 overexpression led to PTEN protein degradation through its direct interaction with HUR. This interaction reduced HUR binding to ubiquitin-specific peptidase 8 (USP8) mRNA, causing degradation of USP8 mRNA and a subsequent decrease in USP8 protein expression. The downregulation of USP8, in turn, increased PTEN polyubiquitination and degradation, culminating in cell malignant transformation and BMIBC anchorageindependent growth. In vivo studies confirmed the downregulation of PTEN and USP8, as well as their positive correlations in both BBN-treated mouse bladder urothelium and tumor tissues of bladder cancer in nude mice. CONCLUSIONS: Our findings, for the first time, demonstrate that overexpressed SNHG1 competes with USP8 for binding to HUR. This competition attenuates USP8 mRNA stability and protein expression, leading to PTEN protein degradation, consequently, this process drives urothelial cell malignant transformation and fosters BMIBC growth and primary BMIBC formation.

Characteristics and a comparison of the gut microbiota in two frog species at the beginning and end of hibernation.

Season has been suggested to contribute to variation in the gut microbiota of animals. The complicated relationships between amphibians and their gut microbiota and how they change throughout the year require more research. Short-term and long-term hypothermic fasting of amphibians may affect gut microbiota differently; however, these changes have not been explored. In this study, the composition and characteristics of the gut microbiota of Rana amurensis and Rana dybowskii during summer, autumn (short-term fasting) and winter (long-term fasting) were studied by high-throughput Illumina sequencing. Both frog species had higher gut microbiota alpha diversity in summer than autumn and winter, but no significant variations between autumn and spring. The summer, autumn, and spring gut microbiotas of both species differed, as did the autumn and winter microbiomes. In summer, autumn and winter, the dominant phyla in the gut microbiota of both species were Firmicutes, Proteobacteria, Bacteroidetes, and Actinobacteria. All animals have 10 OTUs (>90% of all 52 frogs). Both species had 23 OTUs (>90% of all 28 frogs) in winter, accounting for 47.49 ± 3.84% and 63.17 ± 3.69% of their relative abundance, respectively. PICRUSt2 analysis showed that the predominant functions of the gut microbiota in these two Rana were focused on carbohydrate metabolism, Global and overview maps, Glycan biosynthesis metabolism, membrane transport, and replication and repair, translation. The BugBase analysis estimated that among the seasons in the R. amurensis group, Facultatively_Anaerobic, Forms_Biofilms, Gram_Negative, Gram_Positive, Potentially_Pathogenic were significantly different. However, there was no difference for R. dybowskii. The research will reveal how the gut microbiota of amphibians adapts to environmental changes during hibernation, aid in the conservation of endangered amphibians, particularly those that hibernate, and advance microbiota research by elucidating the role of microbiota under various physiological states and environmental conditions.

The study of double-J stent free mode of flexible ureteroscopy and laser lithotripsy: a single centre experience.

OBJECTIVE: To evaluate the effectiveness and safety of flexible ureteroscopy in the treatment of kidney and upper ureteral calculi under double-J stent free mode. METHODS: Data from patients who underwent flexible ureteroscopy and laser lithotripsy between February 2018 and September 2021 were retrospectively and analysed. Cases were grouped according to pre- or postoperative use of the double-J stent (6 Fr): Post-F group (preoperative double-J stent but no postoperative double-J stent); Pre-F group (no preoperative stenting but with postoperative double-J stent); and Routine group (preoperative and postoperative double-J stenting). RESULTS: A total of 554 patients (390 male and 164 female) were included. The mean operation time was similar between the three groups, with no statistically significant difference. Incidence of grade 0-1 ureteral injury was significantly higher in the Pre-F group versus other groups, but there were no significant between-group differences in other operation-related complications. During follow-up, stent-associated complications were observed in the Pre-F and Routine groups, but not in the Post-F group. Stone clearance rates were similar between all groups at 1, 3 and 6 months following surgery. CONCLUSIONS: Flexible ureteroscopy using double-J stent free mode was found to be safe, feasible and effective in treating renal and upper ureteral calculi.

Impact of caregiver burden on caregiver mental health in patients with esophageal cancer: chain mediating effects of benefit finding and rumination, a cross-sectional study.

Background: Cancer caregivers are the main supporter for the tumor patients, they not only need to provide daily nursing to the patients, but also suffering the pressure from economy, emotion and even family members. The mental health of tumor patient is mostly noticed, while not caregivers. The mental health of caregiver greatly affected the nursing quality and even the treatment outcomes. In the current study, the mechanisms underlying the links between caregiver burden, benefit finding, mental health, and rumination in those caring for people with esophageal cancer were examined. Methods: The study was using a convenience sampling, 166 esophageal cancer patients in 2 general hospitals in Jiangsu Province, China, and caregivers of patients were included after excluded the non-conforming patients. Data were collected using investigator-developed questionnaires, the Benefit Finding Scale (BFS), the Event-Related Rumination Inventory (ERRI), the Zarit Burden Interview (ZBI), and the Hospital Anxiety and Depression Scale (HADS) during May 2020 to December 2020. The results were analyzed by SPSS, and the chain mediating effect was analyzed by the the SPSS PROCESS Macro Model. Results: The study comprised 166 caregivers with an average age of (59.96±11.48) years, most of them were female (85.5%). The ZBI was positive correlated with HADS (r=0.882, P<0.01), and negative correlated with BFS (r=-0.873, P<0.01). Intrusive rumination and deliberate rumination in caregivers were negatively correlated (r=-0.901, P<0.01) and positive correlated (r=0.904, P<0.01) with BFS scores, respectively. Furthermore, research have discovered a chain mediation impact of benefit finding and rumination between caregiver burden and psychological well-being among carers of esophageal cancer patients. Conclusions: The findings of this study imply that benefit finding and rumination are crucial components of the coping strategy used to buffer against negative emotion (such as anxiety and depression). Therefore, the mental health of caregivers should also be noticed, and health care professionals should provide targeted interventions to increase the caregiver's level of benefit finding and promote deliberate ruminative thinking.

Value acquisition, value co-creation: The impact of perceived organic grocerant value on customer engagement behavior through brand trust.

Affected by COVID-19, there is a growing trend toward healthy lifestyles and organic food consumption. The literature on organic foods focuses on the factors that influence buying behavior. A thriving organic business requires both sustained consumption and consumer contributions beyond the purchase-customer engagement behavior. The purpose of this study is to examine the factors that may drive member customers to engage with organic grocerants. This study surveyed 280 Chinese member customers of an organic grocerant to explore how to drive customer engagement behavior. Based on value co-creation theory and the customer engagement literature, this study proposed a "value acquisition-value co-creation" framework to explore the relationship between perceived value, brand trust, and customer engagement behavior. The results show that emotional and social value can directly and effectively motivate customer engagement behavior in organic grocerants. However, consumers' perceived quality value and price value will not directly affect customer engagement behavior but instead indirectly affect it through brand trust. Furthermore, improving the perceived value of emotion, quality and price can strengthen brand trust in organic grocerants. The study confirms that brand trust is critical to organic grocerant and customer engagement. Our findings provide a new perspective for understanding the relationship between the value customers receive from organic food consumption and value co-creation through customer engagement behavior.

Increase consumers' willingness to pay a premium for organic food in restaurants: Explore the role of comparative advertising.

Offering organic food is a new trend in the hospitality industry seeking sustainable competitiveness. Premiums and information barriers impede continued growth in organic consumption. This study aims to explore the role of comparative advertising (CA) in organic food communication. Three empirical studies were used to verify the effect of CA vs. non-comparative advertising (NCA) on consumers' willingness to pay a premium (WTPP) for organic food, examining how benefit appeals (health vs. environmental) and consumers' organic skepticism affects CA. The results indicate that matching CA and health appeals increase consumers' WTPP, while environmental appeals have no significant differences between the CA and NCA groups (Study 1). Information persuasiveness mediates the interaction between CA and benefit appeal on WTPP (Study 2). CA increases WTPP among consumers with high organic skepticism, while the interaction between CA and health appeal is only effective for low skepticism consumers (Study 3). The findings unravel and explain the mechanics of how CA works in organic products, which can help restaurants, retailers and tourist destinations advertise organic food to increase consumers' WTPP.

Changes in the gut microbiota diversity of brown frogs (Rana dybowskii) after an antibiotic bath.

BACKGROUND: Captive amphibians frequently receive antibiotic baths to control bacterial diseases. The potential collateral effect of these antibiotics on the microbiota of frogs is largely unknown. To date, studies have mainly relied on oral administration to examine the effects of antibiotics on the gut microbiota; in contrast, little is known regarding the effects of bath-applied antibiotics on the gut microbiota. The gut microbiota compositions of the gentamicin, recovery, and control groups were compared by Illumina high-throughput sequencing, and the functional profiles were analysed using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt). Furthermore, the relationship between the structure and predicted functional composition of the gut microbiota was determined. RESULTS: The alpha diversity indices were significantly reduced by the gentamicin bath, illustrating that this treatment significantly changed the composition of the gut microbiota. After 7 days, the gut microbiota of the recovery group was not significantly different from that of the gentamicin group. Forty-four indicator taxa were selected at the genus level, comprising 42 indicators representing the control group and 2 indicators representing the gentamicin and recovery groups. Potential pathogenic bacteria of the genera Aeromonas, Citrobacter, and Chryseobacterium were significantly depleted after the gentamicin bath. There was no significant positive association between the community composition and functional composition of the gut microbiota in the gentamicin or control frogs, indicating that the functional redundancy of the gut bacterial community was high. CONCLUSIONS: Gentamicin significantly changed the structure of the gut microbiota of R. dybowskii, and the gut microbiota exhibited weak resilience. However, the gentamicin bath did not change the functional composition of the gut microbiota of R. dybowskii, and there was no significant correlation between the structural composition and the functional composition of the gut microbiota.

Effects of phosphite as a plant biostimulant on metabolism and stress response for better plant performance in Solanum tuberosum.

Food availability represents a major worldwide concern due to population growth, increased demand, and climate change. Therefore, it is imperative to identify compounds that can improve crop performance. Plant biostimulants have gained prominence because of their potentials to increase germination, productivity and quality of a wide range of horticultural and agronomic crops. Phosphite (Phi), an analog of orthophosphate, is an emerging biostimulant used in horticulture and agronomy. The aim of this study was to uncover the molecular mechanisms through which Phi acts as a biostimulant with potential effects of overall plant growth. Field and greenhouse experiments, using 4 potato cultivars, showed that following Phi applications, plant performance, including several physio-biochemical traits, crop productivity, and quality traits, were significantly improved. RNA sequencing of control and Phi-treated plants of cultivar Xingjia No. 2, at 0 h, 6 h, 24 h, 48 h, 72 h and 96 h after the Phi application for 24 h revealed extensive changes in the gene expression profiles. A total of 2856 differentially expressed genes were identified, suggesting that multiple pathways of primary and secondary metabolism, such as flavonoids biosynthesis, starch and sucrose metabolism, and phenylpropanoid biosynthesis, were strongly influenced by foliar applications of Phi. GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analyses associated with defense responses revealed significant effects of Phi on a plethora of defense mechanisms. These results suggest that Phi acted as a biostimulant by priming the plants, that was, by triggering dynamic changes in gene expression and modulating metabolic fluxes in a way that allowed plants to perform better. Therefore, Phi usage has the potential to improve crop yield and health, alleviating the challenges posed by the need of feeding a growing world population, while minimizing the agricultural impact on human health and environment.

Physiological and transcriptomic responses of water spinach (Ipomoea aquatica) to prolonged heat stress.

BACKGROUND: Water spinach (Ipomoea aquatica) is an important heat-resistant leafy vegetable that can survive under long-time heat stress condition. However, the physiological characteristics and molecular changes in its response to heat stress are poorly understood. RESULTS: In this study the selected water spinach cultivars with different thermo resistance and their physiological response to heat stress were examined. Under prolonged heat stress, plant growth was inhibited in all tested cultivars. This inhibition was accompanied by the reduction of photosynthetic performance. The reactive oxygen species system in terms of superoxide and hydrogen peroxide contents, as well as antioxidant polyphenols, were evaluated. The results showed that prolonged heat stress caused reduced antioxidant capacity, but the role of antioxidant capacity in a prolonged thermotolerance was not predominant. Transcriptomic analysis of the water spinach subjected to heat stress revealed that 4145 transcripts were specifically expressed with 2420 up-regulated and 1725 down-regulated in heat-sensitive and heat-tolerant cultivars treated with 42 °C for 15 days. Enrichment analysis of these differentially expressed genes showed that the main metabolic differences between heat-sensitive and heat-tolerant cultivars were the carbohydrate metabolism and phenylpropanoid biosynthesis. The results of carbohydrate profiles and RT-qPCR also suggested that heat stress altered carbohydrate metabolism and associated changes in transcriptional level of genes involved in sugar transport and metabolic transition. CONCLUSIONS: The prolonged heat stress resulted in a reduced antioxidant capacity while the role of antioxidant capacity in a prolonged thermotolerance of water spinach was not predominant. Transcriptome analysis and the measurement of carbohydrates as well as the gene expression evaluation indicated that the response of the metabolic pathway such as carbohydrate and phenylpropanoid biosynthesis to heat stress may be a key player in thermo resistance.

Genome-wide identification, characterization, and expression analysis related to autotoxicity of the GST gene family in Cucumis melo L.

Glutathione S-transferase (GST) plays an important role in plant resistance to biotic and abiotic stresses. In this paper, the characteristics of melon GST gene family members were analyzed from a genome-wide perspective. Forty-nine GSTs were identified in melon genome, belonging to eight classes. Through the phylogenetic analysis of GST proteins in melon and other plants, it was found that members from the same subfamily in different species clustered together, indicating that the subfamilies of GST have diversified before the divergence within these species. The results of chromosome mapping showed that GSTs were present in all chromosomes except for chromosome 5. Gene replication events played an important role in the expansion and evolution of melon GST gene family. Ten GSTs with significant differential expression were screened in the transcriptome database related to melon autotoxicity stress. The differential expression of these 10 GSTs was detected in roots and leaves of melon seedlings treated with cinnamic acid. The relative expression level of CmGSTU7, CmGSTU10, CmGSTU18, CmGSTF2 and CmGSTL1 in roots of melon seedlings was significantly higher than that in control group. It suggested that the five GSTs might play an important role in cinnamic acid mediated autotoxicity stress in melon. The results of this paper were helpful to reveal the evolution and functional succession of GST family and further understand the response of GST to autotoxicity stress in melon.

Specific response mechanism to autotoxicity in melon (Cucumis melo L.) root revealed by physiological analyses combined with transcriptome profiling.

Melon is of great value in food, medicine and industry. In recent years, the continuous cropping obstacles of melon is increasingly prominent, which seriously affects the cultivation. Autotoxicity is the key factor for the obstacles. Root is the first line against autotoxicity and main organs for autotoxins secretion. Some physiological responses and differentially expressed genes (DEGs) related to autotoxicity are only limited to root system. Considering the lack of relevant research, physiological researches combined with transcriptome sequencing of melon seedling after autotoxicity stress mediated by root exudates (RE) was performed to help characterize the response mechanism to autotoxicity in melon roots. The results showed that autotoxicity inhibited root morphogenesis of melon seedlings, induced the excessive accumulation of reactive oxygen species (ROS) and lipid peroxidation in roots, and activated most antioxidant enzymes. Compared with the control group, the osmoregulation substance content was always at a high level. DEGs response to autotoxicity in roots were distinguished from that in leaves. Functional annotation of these DEGs suggested that autotoxicity affected biological regulation in a negative manner. DEGs were mainly involved in the synthesis of antioxidants, DNA damage and metabolism, and stress response. These setbacks were associated with the deterioration of root morphogenesis, generation of dwarf and slender roots, and ultimately leading to plant death. The results may provide important information for revealing the response mechanism of root to autotoxicity, and provide theoretical basis for solving the continuous cropping obstacles in melon.

Isolation and hypoglycemic effects of water extracts from mulberry leaves in Northeast China.

Diabetes is the main chronic disease that greatly affects human life. Up to now, many measures have been taken to cope with the disease, among which natural products with hypoglycemic effects have aroused great interest. The objective of this study was to evaluate the hypoglycemic effects of Morus abla L. cv. longsang 1 leaf-derived water extract in vitro and in vivo. These leaves were firstly subjected to water extraction, and the obtained products were further isolated for polysaccharides, flavonoids and alkaloids. The α-glucosidase activity and anti-protein glycosylation activity of the aqueous extracts were examined in vitro. Hyperglycemic mouse models were used to evaluate the hypoglycemic effects of the aqueous extract by blood biochemical parameters, intestinal microbiota, and pathological changes to the kidneys. The results showed that the main hypoglycemic components in the aqueous extracts were flavonoids and alkaloids and their inhibition rates against α-glucosidase activity were 86.12 ± 1.79% and 87.29 ± 1.32%, respectively. High-dose mulberry leaf water extracts can reduce the blood glucose of diabetic mice by 28.17% and improve glucose tolerance by 19.02%. Furthermore, mulberry leaf water extracts could reduce the serum free fatty acid (FFA), tumor necrosis factor-α (TNF-α), insulin and glycated serum protein content, while alleviating kidney damage and improving intestinal microbiota. These results indicated that the synergistic effects among the different components of mulberry leaves might explain their alleviating effects on diabetic syndrome and thus provide a simple, convenient way to obtain the hypoglycemic components from mulberry leaves.

Phosphite Application Alleviates Pythophthora infestans by Modulation of Photosynthetic and Physio-Biochemical Metabolites in Potato Leaves.

Potato late blight (Phytophtora infestans) is among the most severely damaging diseases of potato (Solanum tuberusom L.) worldwide, causing serious damages in potato leaves and tubers. In the present study, the effects of potassium phosphite (KPhi) applications on photosynthetic parameters, enzymatic and non-enzymatic antioxidant properties, hydrogen peroxide (H2O2) and malondialdehyde (MDA), total protein and total carbohydrate of potato leaves challenged with P. infestans pathogen were investigated. Potato leaves were sprayed five times with KPhi (0.5%) during the growing season prior to inoculation with P. infestans. The potato leaves were artificially infected by the LC06-44 pathogen isolate. The leaves were sampled at 0, 24, 48, 72 and 96 h after the infection for evaluations. P. infestans infection reduced chlorophyll (Chl) pigments contents, chlorophyll fluorescence, carotenoid (Car) and anthocyanin contents and increased the accumulation of H2O2 and MDA. Meanwhile, our result showed that KPhi treatment alleviated adverse effect of late blight in potato leaves. KPhi application also increased plant tolerance to the pathogen with improved photosynthetic parameters Chl a, b, total Chl, Car, and anthocyanin compare to controls. Moreover, the increased oxidative enzymes activity of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APx), and non-enzymatic substances such as phenolics, flavonoids and proline were found in KPhi treated plants, compared to untreated plants after inoculation. In addition, KPhi application followed by P. infestans infection also decreased the content of H2O2 and MDA, but increased the total protein and total carbohydrate contents in potato leaves. The consequence of current research indicated that KPhi played a vital role in pathogen tolerance, protecting the functions of photosynthetic apparatus by improved oxidative levels and physio-biochemical compounds in potato leaves.

Exogenous phosphite application alleviates the adverse effects of heat stress and improves thermotolerance of potato (Solanum tuberosum L.) seedlings.

Phosphite (Phi), an analog of phosphate (Pi) anion, is emerging as a potential biostimulator, fungicide and insecticide. Here, we reported that Phi also significantly enhanced thermotolerance in potatoes under heat stress. Potato plants with and without Phi pretreatment were exposed to heat stress and their heat tolerance was examined by assessing the morphological characteristics, photosynthetic pigment content, photosystem II (PS II) efficiency, levels of oxidative stress, and level of DNA damage. In addition, RNA-sequencing (RNA-Seq) was adopted to investigate the roles of Phi signals and the underlying heat resistance mechanism. RNA-Seq revealed that Phi orchestrated plant immune responses against heat stress by reprograming global gene expressions. Results from physiological data combined with RNA-Seq suggested that the supply of Phi not only was essential for the better plant performance, but also improved thermotolerance of the plants by alleviating oxidative stress and DNA damage, and improved biosynthesis of osmolytes and defense metabolites when exposed to unfavorable thermal conditions. This is the first study to explore the role of Phi in thermotolerance in plants, and the work can be applied to other crops under the challenging environment.

Prevalence of cardiovascular risk factors in non-menopausal and postmenopausal inpatients with type 2 diabetes mellitus in China.

BACKGROUND: To investigate the prevalence of cardiovascular disease (CVD) risk factors and assess the 10-year risk of CVD in non-menopausal and postmenopausal women with type 2 diabetes mellitus (T2DM). METHODS: A total of 569 patients with T2DM at a Chinese tertiary hospital were investigated using the Framingham Risk Score (FRS). We evaluated the 10-year risk of CVD, clinical and menopause characteristics in all subjects. RESULTS: Among the 569 diabetic patients, the incidence of smoking, dyslipidemia, hypertension, overweight or obesity, and nonalcoholic fatty liver disease (NAFLD) was 0.7, 36.2, 38.1 56.6 and 58.2%, respectively. The usage rate of hypoglycemic agents, antihypertensive agents, lipid modulators and antithrombotic drugs was 88.6, 78.3, 50.0 and 27.1%, respectively. However, only 1.2% of inpatients achieved the three target goals for the control of blood glucose (HbA1c < 7%), blood pressure (systolic blood pressure < 130 mmHg, diastolic blood pressure < 80 mmHg), and blood lipids (total cholesterol < 174 mg/dL). The 10-year risk of CVD was (1.6 ± 1.5%) and tended to increase along with age (F = 27.726, P <  0.001). For all subjects (n = 569), multiple linear regression analysis showed that menopause (ß = 0.275, P <  0.001), low-density lipoprotein cholesterol (LDL-C) (ß = 0.212, P <  0.001), fasting plasma glucose (FPG) (ß = 0.093, P = 0.018) and waist-to-hip-ratio (ß = - 0.078, P = 0.047) were risk factors of 10-year risk of CVD, which may explain the variance of 14.3%. In the postmenopausal group (n = 397), LDL-C (ß = 0.227, P <  0.001), FPG (ß = 0.139, P = 0.003) and time since menopause (ß = 0.230, P <  0.001) were found to be associated with CVD, which may explain the variance of 14.6%. CONCLUSION: The incidence of dyslipidmia, hypertension, overweight or obesity and NAFLD is high. The level of control of blood glucose, blood pressure, and blood lipids was found to be extremely low and the treatment status was not ideal. Besides menopause, LDL-C, FPG and time since menopause were found to be independent risk factors for the 10-year risk of CVD. Therefore, it is necessary to focus on comprehensive control of multiple risk factors, such as plasma glucose, blood pressure and serum lipid.

siRNA-mediated NCAM1 gene silencing suppresses oxidative stress in pre-eclampsia by inhibiting the p38MAPK signaling pathway.

Pre-eclampsia (PE), whose pathophysiology and etiology remain undefined, represents a leading consequence of fetal and maternal mortality and morbidity. Oxidative stress (OS) is recognized to involve in this disorder. In this study, we hypothesized that neural cell adhesion molecule 1 (NCAM1) gene silencing would suppress the OS in the pregnancy complicated by PE. Initially, clinical samples were collected for determination of NCAM1 expression in placental tissues and levels of OS products in blood. To assess the regulatory mechanism of NCAM1 knockdown on OS, we used small interfering RNA (siRNA) to silence NCAM1 expression in human umbilical vein endothelial cells (HUVECs). Next, cells were treated with or without hypoxia/reoxygenation to observe the level changes of OS products and p38 mitogen-activated protein kinase (p38MAPK) pathway-related genes. Finally, an evaluation of HUVEC migration and invasion abilities was conducted by wound-healing and transwell assays. Placenta of pregnancy with PE presented significantly increased NCAM1 expression in comparison to placenta of normal pregnancy. Meanwhile, enhanced OS in blood of pregnant women with PE was observed relative to women with normal pregnancy. siRNA-mediated knockdown of NCAM1 gene could inhibit the p38MAPK signaling pathway, repress OS, and promote cell migration and invasion in HUVECs, indicating that NCAM1 inhibition could reduce the influence of PE. Importantly, blocking the p38MAPK signaling pathway reversed the inhibitory role of NCAM1 gene silencing on PE. Collectively, this study defines potential role of NCAM1 gene silencing as a therapeutic target in PE through inhibiting OS and enhancing HUVEC migration and invasion by disrupting the p38MAPK signaling pathway.

Comparative transcriptome analyses revealed different heat stress responses in high- and low-GS Brassica alboglabra sprouts.

BACKGROUND: Chinese kale (Brassica alboglabra) contains high nutritional elements and functional molecules, especially anticarcinogenic and antioxidant glucosinolates (GS), which was highly affected by environment temperature. To investigate the link of GS biosynthesis with heat stress response in Chinese kale, global transcription profiles of high-GS line (HG), low-GS line (LG), high-GS line under heat stress (HGT) and low-GS line under heat stress (LGT) were analyzed. RESULTS: Based on three biological replicates of each RNA sequencing data, 3901, 4062 and 2396 differentially expressed genes in HG vs HGT, LG vs LGT and HGT vs LGT were obtained, respectively. GO annotation, KEGG pathway analysis and a comprehensive analysis of DEGs showed a strong correlation between the GS biosynthesis and heat stress response. It was noticed that 11 differentially expressed genes tied to the GS biosynthesis were down-regulated, 23 heat shock transcription factors and 61 heat shock proteins were up-regulated upon the heat treatment. Another two Chinese kale varieties Cuibao and Shunbao with high- and low- GS content respectively, were used to validate the relationship of GS content and heat-response, and the results showed that high-GS content variety were more thermotolerant than the low-GS content one although GS significantly decreased in both varieties under heat stress. In addition, HSP100/ClpB, HSP90, HSP70 and sHSPs were differentially expressed in high- and low-GS varieties. Notably, HSP90 and sHSPs showed an obviously early response to heat stress than other related genes. CONCLUSION: The higher heat resistance of high-GS Chinese kale and the sharp decrease of glucosinolate content under heat stress indicated a strong relationship of GS accumulation and heat stress response. Combined with the previous report on the low expression of HSP90 at elevated temperatures in GS-deficient mutant TU8 of Arabidopsis, the differential expression pattern of HSP90 in high- and low- GS varieties and its early heat response implied it might be a key regulator in GS metabolism and heat-resistance in Chinese kale.

Selection of Reference Genes for Real-time Quantitative PCR Normalization in the Process of Gaeumannomyces graminis var. tritici Infecting Wheat.

Gaeumannomyces graminis var. tritici is a soil borne pathogenic fungus associated with wheat roots. The accurate quantification of gene expression during the process of infection might be helpful to understand the pathogenic molecular mechanism. However, this method requires suitable reference genes for transcript normalization. In this study, nine candidate reference genes were chosen, and the specificity of the primers were investigated by melting curves of PCR products. The expression stability of these nine candidates was determined with three programs-geNorm, Norm Finder, and Best Keeper. TUBß was identified as the most stable reference gene. Furthermore, the exopolygalacturonase gene (ExoPG) was selected to verify the reliability of TUBß expression. The expression profile of ExoPG assessed using TUBß agreed with the results of digital gene expression analysis by RNA-Seq. This study is the first systematic exploration of the optimal reference genes in the infection process of Gaeumannomyces graminis var. tritici.