Genome-wide identification and expression analysis of the ADH gene family under diverse stresses in tobacco (Nicotiana tabacum L.).

BACKGROUND: Alcohol dehydrogenases (ADHs) are the crucial enzymes that can convert ethanol into acetaldehyde. In tobacco, members of ADH gene family are involved in various stresses tolerance reactions, lipid metabolism and pathways related to plant development. It will be of great application significance to analyze the ADH gene family and expression profile under various stresses in tobacco. RESULTS: A total of 53 ADH genes were identified in tobacco (Nicotiana tabacum L.) genome and were grouped into 6 subfamilies based on phylogenetic analysis. Gene structure (exon/intron) and protein motifs were highly conserved among the NtADH genes, especially the members within the same subfamily. A total of 5 gene pairs of tandem duplication, and 3 gene pairs of segmental duplication were identified based on the analysis of gene duplication events. Cis-regulatory elements of the NtADH promoters participated in cell development, plant hormones, environmental stress, and light responsiveness. The analysis of expression profile showed that NtADH genes were widely expressed in topping stress and leaf senescence. However, the expression patterns of different members appeared to be diverse. The qRT-PCR analysis of 13 NtADH genes displayed their differential expression pattern in response to the bacterial pathogen Ralstonia solanacearum L. INFECTION: Metabolomics analysis revealed that NtADH genes were primarily associated with carbohydrate metabolism, and moreover, four NtADH genes (NtADH20/24/48/51) were notably involved in the pathway of alpha-linolenic acid metabolism which related to the up-regulation of 9-hydroxy-12-oxo-10(E), 15(Z)-octadecadienoic acid and 9-hydroxy-12-oxo-15(Z)-octadecenoic acid. CONCLUSION: The genome-wide identification, evolutionary analysis, expression profiling, and exploration of related metabolites and metabolic pathways associated with NtADH genes have yielded valuable insights into the roles of these genes in response to various stresses. Our results could provide a basis for functional analysis of NtADH gene family under stressful conditions.

Functional and evolutionary comparative analysis of the DIR gene family in Nicotiana tabacum L. and Solanum tuberosum L.

BACKGROUND: The dirigent (DIR) genes encode proteins that act as crucial regulators of plant lignin biosynthesis. In Solanaceae species, members of the DIR gene family are intricately related to plant growth and development, playing a key role in responding to various biotic and abiotic stresses. It will be of great application significance to analyze the DIR gene family and expression profile under various pathogen stresses in Solanaceae species. RESULTS: A total of 57 tobacco NtDIRs and 33 potato StDIRs were identified based on their respective genome sequences. Phylogenetic analysis of DIR genes in tobacco, potato, eggplant and Arabidopsis thaliana revealed three distinct subgroups (DIR-a, DIR-b/d and DIR-e). Gene structure and conserved motif analysis showed that a high degree of conservation in both exon/intron organization and protein motifs among tobacco and potato DIR genes, especially within members of the same subfamily. Total 8 pairs of tandem duplication genes (3 pairs in tobacco, 5 pairs in potato) and 13 pairs of segmental duplication genes (6 pairs in tobacco, 7 pairs in potato) were identified based on the analysis of gene duplication events. Cis-regulatory elements of the DIR promoters participated in hormone response, stress responses, circadian control, endosperm expression, and meristem expression. Transcriptomic data analysis under biotic stress revealed diverse response patterns among DIR gene family members to pathogens, indicating their functional divergence. After 96 h post-inoculation with Ralstonia solanacearum L. (Ras), tobacco seedlings exhibited typical symptoms of tobacco bacterial wilt. The qRT-PCR analysis of 11 selected NtDIR genes displayed differential expression pattern in response to the bacterial pathogen Ras infection. Using line 392278 of potato as material, typical symptoms of potato late blight manifested on the seedling leaves under Phytophthora infestans infection. The qRT-PCR analysis of 5 selected StDIR genes showed up-regulation in response to pathogen infection. Notably, three clustered genes (NtDIR2, NtDIR4, StDIR3) exhibited a robust response to pathogen infection, highlighting their essential roles in disease resistance. CONCLUSION: The genome-wide identification, evolutionary analysis, and expression profiling of DIR genes in response to various pathogen infection in tobacco and potato have provided valuable insights into the roles of these genes under various stress conditions. Our results could provide a basis for further functional analysis of the DIR gene family under pathogen infection conditions.

Coping target checklist for home-based older adults living with disabilities and their spousal caregivers.

BACKGROUND: The substantial rise in the population of older adults living with disabilities is a prominent concern, presenting a profound challenge for healthcare and social welfare systems. Community-based home care is seen as an effective approach to meet the care needs of older adults living with disabilities. OBJECTIVE: To construct a coping target checklist for home-based older adults living with disabilities and their spousal caregivers. METHODS: The initial draft was developed based on a comprehensive literature review, followed by two rounds of Delphi correspondence final version. RESULTS: A comprehensive literature review resulted in the development of 7 modules, 20 topics. After round 1, 3 items were removed, 3 sections, 1 topic and 1 objective were new additions, 16 items were modified, split or combined. Four sections, 3 sections (Individual coping target for spousal caregivers, Individual coping target for older adults living with disabilities, and Shared coping target), 7 modules, 18 topics and 49 objectives were finally identified in round 2. The content of the list tool is derived from three perspectives: self-management strategies for older adults living with disabilities, caregiving strategies for spousal caregivers, and combined. CONCLUSIONS: The coping target checklist was intended to be evidence-based and reflective of a practical direction for home-based older adults living with disabilities and their spousal caregivers living at home.

Health empowerment and health-related quality of life in older individuals with disabilities and their spouse carers: A cross-sectional study.

The stress of disability significantly impacts an individual's quality of life and that of a spouse. Health empowerment, based on the idea that individuals may be successful despite disability, may be meaningful to disabled persons and their spouse carers. This cross-sectional survey study aimed to explore the effect of health empowerment on the health-related quality of life (HRQOL) of older individuals with disabilities and their spouse carers on both a personal (actor effect) and interpersonal level (partner effect). A total of 1092 dyads of older individuals with disabilities and their spouse carers residing in communities were recruited from seven provinces in China. Two separate Actor-Partner Interdependence Model analyses were conducted to examine the impact of health empowerment on the two domains of HRQOL: the Physical Component Score (PCS) and the Mental Component Score (MCS). The results revealed that health empowerment had actor effects on the PCS and MCS of older individuals with disabilities, as well as on the PCS and MCS of their spouse carers. However, no significant partner effects of health empowerment on the PCS and MCS of either the individuals with disabilities or their spouse caregivers were observed. Empowering individuals with disabilities and their spouse carers may help them enhance their own HRQOL, both physically and mentally. However, more research is required to determine the interpersonal effect of health empowerment on the HRQOL.

Overexpression of MdZAT5, an C2H2-Type Zinc Finger Protein, Regulates Anthocyanin Accumulation and Salt Stress Response in Apple Calli and Arabidopsis.

Zinc finger proteins are widely involved and play an important role in plant growth and abiotic stress. In this research, MdZAT5, a gene encoding C2H2-type zinc finger protein, was cloned and investigated. The MdZAT5 was highly expressed in flower tissues by qRT-PCR analyses and GUS staining. Promoter analysis showed that MdZAT5 contained multiple response elements, and the expression levels of MdZAT5 were induced by various abiotic stress treatments. Overexpression of MdZAT5 in apple calli positively regulated anthocyanin accumulation by activating the expressions of anthocyanin biosynthesis-related genes. Overexpression of MdZAT5 in Arabidopsis also enhanced the accumulation of anthocyanin. In addition, MdZAT5 increased the sensitivity to salt stress in apple calli. Ectopic expression of MdZAT5 in Arabidopsis reduced the expression of salt-stress-related genes (AtNHX1 and AtABI1) and improved the sensitivity to salt stress. In conclusion, these results suggest that MdZAT5 plays a positive regulatory role in anthocyanin accumulation and negatively regulates salt resistance.

A super-hydrophilic partially reduced graphene oxide membrane with improved stability and antibacterial properties.

In order to improve stability and antibacterial property, a novel super-hydrophilic partially reduced graphene oxide membrane was prepared by interfacial polymerization of piperazine and partially reduced graphene oxide as aqueous solution and trimesoyl chloride as organic solution. Fourier transform infrared spectroscopy, scanning electron microscope, and contact angle measurement were conducted to probe the morphology and properties of the membranes. The modified membrane possessed super-hydrophilicity, improved durability and swelling resistance. The optimized membrane had a molecular weight cut off of about 674 Da and possessed a pure water permeability of 49.86 L·m-2·h-1·MPa-1. The retention order of salts was Na2SO4 > MgSO4 > MgCl2 > Na2CO3 > CaCl2 > NaCl, while the rejection for four kinds of pharmaceuticals followed the order of ibuprofen (92%) > carbamazepine (87%) > amlodipine (80%) > atenolol (76%), indicating that the negatively charged membrane could improve the retention performance by the electrostatic repulsive effect. Moreover, the enhanced antibacterial performance of membrane attributed to the dual effects of the super-hydrophilicity and the tea polyphenols antibacterial material loading, which may alter the charge distribution on and within the membrane, leading to loss of cell viability.

A high stability GO nanofiltration membrane preparation by co-deposition and crosslinking polydopamine for rejecting dyes.

In order to improve the stability of nanofiltration membrane in separation and purification, a novel polyelectrolyte multilayer nanofiltration membrane was facilely prepared by co-deposition of polydopamine (PDA) and polyethyleneimine (PEI) on the polyethersulfone (PES) ultrafiltration membrane substrate, followed by immersing graphene oxide (GO) solution, and crosslinking PDA. The modified surfaces were characterized using scanning electron microscopy (SEM) and atomic force microscopy (AFM), water contact angle, their saline flux and ability to reject salt and dye were determined. The results also exhibited salt rejection ability as Na2SO4 > K2SO4 > MgSO4 > NaCl > KCl > MgCl2, suggesting the higher rejection of divalent anion. Also, the retention order of the dye by the GO modified membrane is DY86 > DB19 > AG27 > DY142 > DB56 > AR151 > VB5, indicating that the GO modified membrane has better rejection of negatively charged dyes as well as higher molecular weight dyes. Ethanol and hypochlorite resistance tests under different pH conditions showed the membranes coated GO enhanced stability in regard to salt rejection properties. Significantly, the anti-biological test confirmed the growth rate of microalgae on the GO introduced membrane was decreased greatly due to enhanced stability and lower roughness.

Thalidomide targets EGFL6 to inhibit EGFL6/PAX6 axis-driven angiogenesis in small bowel vascular malformation.

BACKGROUND: Small bowel vascular malformation disease (SBVM) is the most common cause of obscure gastrointestinal bleeding (OGIB). Several studies suggested that EGFL6 was able to promote the growth of tumor endothelial cells by forming tumor vessels. To date, it remains unclear how EGFL6 promotes pathological angiogenesis in SBVM and whether EGFL6 is a target of thalidomide. METHODS: We took advantage of SBVM plasma and tissue samples and compared the expression of EGFL6 between SBVM patients and healthy people via ELISA and Immunohistochemistry. We elucidated the underlying function of EGFL6 in SBVM in vitro and by generating a zebrafish model that overexpresses EGFL6, The cycloheximide (CHX)-chase experiment and CoIP assays were conducted to demonstrate that thalidomide can promote the degradation of EGFL6 by targeting CRBN. RESULTS: The analysis of SBVM plasma and tissue samples revealed that EGFL6 was overexpressed in the patients compared to healthy people. Using in vitro and in vivo assays, we demonstrated that an EMT pathway triggered by the EGFL6/PAX6 axis is involved in the pathogenesis of SBVM. Furthermore, through in vitro and in vivo assays, we elucidated that thalidomide can function as anti-angiogenesis medicine through the regulation of EGFL6 in a proteasome-dependent manner. Finally, we found that CRBN can mediate the effect of thalidomide on EGFL6 expression and that the CRBN protein interacts with EGFL6 via a Lon N-terminal peptide. CONCLUSION: Our findings revealed a key role for EGFL6 in SBVM pathogenesis and provided a mechanism explaining why thalidomide can cure small bowel bleeding resulting from SBVM.

miR-194 inhibits liver cancer stem cell expansion by regulating RAC1 pathway.

Liver cancer stem cells (CSCs) contribute to tumorigenesis, progression, drug resistance and recurrence of hepatocellular carcinoma (HCC). However, the underlying mechanism for the propagation of liver CSCs remains unclear. Herein, we observed low expression of miR-194 in chemoresistant HCC cells. A remarkable decrease of miR-194 was detected in EpCAM or CD133-positive liver CSCs and CSC-enriched hepatoma spheres. Interference miR-194 facilitated liver CSCs expansion by enhancing the self-renewal of liver CSCs. While up-regulating miR-194 inhibited liver CSCs expansion by suppressing the self-renewal of liver CSCs. Furthermore, hepatoma cells with miR-194 overexpression performed more sensitivity to sorafenib treatment. Mechanistically, functional studies found that Ras-related C3 botulinum toxin substrate 1 (RAC1) was a direct target of miR-194. Overexpression of miR-194 inhibited the expression of RAC1 in liver CSCs. Special RAC1 siRNA diminished the discrepancy in liver CSC proportion and the self-renewal capacity between miR-194 overexpression hepatoma cells and control cells, which further confirmed that RAC1 was required in miR-194-inhibited liver CSCs expansion. More importantly, downregulated expression of miR-194 was a predictor of poor prognosis of HCC patients.

EGFL6 promotes cell proliferation in colorectal cancer via regulation of the WNT/ß-catenin pathway.

Epidermal growth factor-like protein 6 (EGFL6) serves as an exocrine protein promoting proliferation and migration during carcinogenesis in ovarian cancer. However, its function and mechanisms in colorectal cancer (CRC) have not been completely explored. To investigate the role of EGFL6 in CRC cell growth, in vitro CCK8, colony formation assays, flow cytometric analysis of the cell cycle and apoptosis, and an in vivo tumor xenograft model were utilized. Additionally, Western blotting and luciferase reporter assays were used to investigate the underlying mechanisms of EGFL6 function on the WNT/ß-catenin signaling pathway. Immunohistochemical staining showed that EGFL6 is overexpressed in CRCs and this overexpression was highly correlated with advanced T classification, N classification, distant metastasis, and poor survival. Knocking down EGFL6 in CRC cell lines induced the inhibition of cell growth, cell cycle arrest at the G0/G1 phase, and apoptosis. Further, knockdown of EGFL6 blocked WNT/ß-catenin signaling as measured by Western blotting and luciferase reporter assay. Results also showed that recombinant EGFL6 (rEGFL6) induced ß-catenin in a concentration- and time-dependent manner. Further experiments showed that administration of rEGFL6 to cell cultures with EGFL6 knocked down or treated with the WNT/ß-catenin inhibitor ICG-001 increased ß-catenin and its downstream protein CyclinD1. The CCK8 assay showed that EGFL6 promoted CRC cell growth partly by the promotion of TCF7L2 expression. These findings suggest that EGFL6 plays a crucial role in the progression of CRC by regulation of the WNT/ß-catenin signaling pathway.

Pseudopod-associated protein KIF20B promotes Gli1-induced epithelial-mesenchymal transition modulated by pseudopodial actin dynamic in human colorectal cancer.

Kinesin family member 20B (KIF20B) has been reported to have an oncogenic role in bladder and hepatocellular cancer cells, but its role in colorectal cancer (CRC) progression remains unclear. In this study, we assessed the mRNA and protein levels of KIF20B in CRC tissues using qRT-PCR and immunohistochemistry, respectively. KIF20B was overexpressed in CRC tissues and was associated with cancer invasion and metastasis. Mechanistically, KIF20B overexpression promoted the epithelial-mesenchymal transition (EMT) process mediated by glioma-associated oncogene 1 (Gli1) as well as CRC cell migration and invasion. Interestingly, KIF20B was localized in pseudopod protrusions of CRC cells and influenced the formation of cell protrusions, especially the EMT-related invadopodia. Moreover, intracellular actin dynamic participated in the modulation of the Gli1-mediated EMT and EMT-related cell pseudopod protrusion formation induced by KIF20B. We identified a role for KIF20B in CRC progression and revealed a correlation between KIF20B expression in CRC tissues and patient prognosis. The underlying mechanism was associated with the Gli1-mediated EMT and EMT-related cell protrusion formation modulated by intracellular actin dynamic. Thus, KIF20B may be a potential biomarker and promising treatment target for CRC.

[Research on On-Line Calibration Based Photoacoustic Spectrometry System for Monitoring the Concentration of CO2 in Atmosphere].

Resonate frequency and cell constant of photoacoustic spectrum system are usually calibrated by using standard gas in laboratory, whereas the resonate frequency and cell constant will be changed in-situ, leading to measurement accuracy errors, caused by uncertainties of standard gas, differences between standard and measured gas components and changes in environmental condition, such as temperature and humidity. As to overcome the above problems, we have proposed an on-line atmospheric oxygen-based calibration technology for photoacoustic spectrum system and used in measurement of concentration of carbon dioxide in atmosphere. As the concentration of atmospheric oxygen is kept as constant as 20.96%, the on-line calibration for the photoacoustic spectrum system can be realized by detecting the swept-frequency and peak signal at 763.73 nm. The cell of the PAS has a cavity with length of 100 mm and an inner diameter of 6 mm, and worked in a first longitudinal resonant mode. The influence of environmental temperature and humidity, gas components on the photoacoustic cell's performance has been theoretically analyzed, and meanwhile the resonant frequencies and cell constants were calibrated and acquired respectively using standard gas, indoor air and outdoor air. Compared with calibrated gas analyzer, concentration of carbon dioxide is more accurate by using the resonant frequency and cell constant calculated by oxygen in tested air, of which the relative error is less than 1%, much smaller than that calculated by the standard gas in laboratory. The innovation of this paper is that using atmospheric oxygen as photoacoustic spectrum system's calibration gas effectively reduces the error caused by using standard gas and environmental condition changes, and thus improves the on-line measuring accuracy and reliability of the photoacoustic spectrum system.

Accuracy and influencing factors of Type B2 vessels in predicting the invasion depth of superficial esophageal squamous cell carcinoma under narrow-band imaging magnifying endoscopy.

OBJECTIVE: To evaluate the accuracy of Type B2 based on narrow-band imaging-magnifying endoscopy (NBI-ME) in judging invasion depth of superficial esophageal squamous cell carcinoma (SESCC) and analyze potential influencing factors. METHOD: Data from 113 patients where Type B2 was observed by magnifying endoscopy and confirmed by postoperative pathology as SESCC at the First Affiliated Hospital of Fujian Medical University and Fujian Provincial Hospital from January 2015 to April 2024 were retrospectively analyzed. Patients were divided into correct prediction and incorrect prediction groups according to the postoperative pathological results, and the prediction accuracy was calculated. The incorrect prediction group was further divided into overestimation and underestimation groups to identify the influential factors for overprediction and underprediction, respectively. The independent influential factors for the prediction were assessed using multivariate Cox logistic regression analysis. RESULTS: B2-narrow (Type B2 area ≤ 5 mm) (p < 0.001) and Type B2 around erosion (p = 0.040) were independent risk factors of overpredicting the invasion depth of SESCC based on Type B2. The presence of significant endoscopic features was an independent protective factor for overpredicting the invasion depth of SESCC (p = 0.014), whereas the presence of significant features under endoscopy was an independent risk factor for the underprediction (p = 0.005). CONCLUSION: B2-narrow (Type B2 area ≤ 5 mm), Type B2 vessels around erosion, and non- significant endoscopic features are closely related to overpredicting the invasion depth of SESCC based on Type B2, and the presence of significant endoscopic features is closely associated with underprediction.