Enhanced anticancer efficacy of TRAIL-conjugated and odanacatib-loaded PLGA nanoparticles in TRAIL resistant cancer.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) demonstrates unique characteristics in anticancer therapies as it selectively induces apoptosis in cancer cells. However, most cancer cells are TRAIL-resistant. Odanacatib (ODN), a cathepsin K inhibitor, is considered a novel sensitizer for cancer treatment. Combination therapy between TRAIL and sensitizers is considered a potent platform that improves TRAIL-based anticancer therapies beyond TRAIL monotherapy. Herein, we developed ODN loaded poly(lactic-co-glycolic) nanoparticles conjugated to GST-TRAIL (TRAIL-ODN-PLGA-NPs) to target and treat TRAIL-resistant cancer. TRAIL-ODN-PLGA-NPs demonstrated a significant increase in cellular uptake via death receptors (DR5 and DR4) on surface of cancer cells. TRAIL-ODN-PLGA-NPs exposure destroyed more TRAIL-resistant cells compared to a single treatment with free drugs. The released ODN decreased the Raptor protein, thereby increasing damage to mitochondria by elevating reactive oxygen species (ROS) generation. Additionally, Bim protein stabilization improved TRAIL-resistant cell sensitization to TRAIL-induced apoptosis. The in vivo biodistribution study revealed that TRAIL-ODN-PLGA-NPs demonstrated high location and retention in tumor sites via the intravenous route. Furthermore, TRAIL-ODN-PLGA-NPs significantly inhibited xenograft tumor models of TRAIL-resistant Caki-1 and TRAIL-sensitive MDA-MB-231 cells.The inhibition was associated with apoptosis activation, Raptor protein stabilizing Bim protein downregulation, Bax accumulation, and mitochondrial ROS generation elevation. Additionally, TRAIL-ODN-PLGA-NPs affected the tumor microenvironment by increasing tumor necrosis factor-α and reducing interleukin-6. In conclusion, we evealed that our formulation demonstrated synergistic effects against TRAIL compared with the combination of free drug in vitro and in vivo models. Therefore, TRAIL-ODN-PLGA-NPs may be a novel candidate for TRAIL-induced apoptosis in cancer treatment.

Characteristics and phytotoxicity of hydrochar-derived dissolved organic matter: Effects of feedstock type and hydrothermal temperature.

The dissolved organic matter (DOM) with high mobility and reactivity plays a crucial role in soil. In this study, the characteristics and phytotoxicity of DOM released from the hydrochars prepared from different feedstocks (cow manure, corn stalk and Myriophyllum aquaticum) under three hydrothermal carbonization (HTC) temperatures (180, 200 and 220°C) were evaluated. The results showed that the hydrochars had high dissolved organic carbon content (20.15 to 37.65 mg/g) and its content showed a gradual reduction as HTC temperature increased. Three fluorescent components including mixed substance of fulvic acid-like and humic acid-like substances (C1, 30.92%-58.32%), UVA humic acid-like substance (C2, 25.27%-29.94%) and protein-like substance (C3, 11.74%-41.92%) were identified in hydrochar DOM by excitation emission matrix spectra coupled with parallel factor analysis. High HTC temperature increased the relative proportion of aromatic substances (C1+C2) and humification degree of hydrochar DOM from cow manure, while it presented adverse effects on the hydrochar DOM from corn stalk and Myriophyllum. aquaticum. The principal component analysis suggested that feedstock type and HTC temperature posed significant effects on the characteristics of hydrochar DOM. Additionally, seed germination test of all hydrochar DOM demonstrated that the root length was reduced by 8.88%-26.43% in contrast with control, and the germination index values were 73.57%-91.12%. These findings provided new insights into the potential environmental effects for hydrochar application in soil.

Unintentional emissions of polychlorinated naphthalenes in China: Sources, composition, and historical trends.

Polychlorinated naphthalenes (PCNs) are detrimental to human health and the environment. With the commercial production of PCNs banned, unintentional releases have emerged as a significant environmental source. However, relevant information is still scarce. In this study, provincial emissions for eight PCNs homologues from 37 sources in the Chinese mainland during the period of 1960-2019 were estimated based on a source-specific and time-varying emission factor database. The results showed that the total PCNs emissions in 2019 reached 757.0 kg with Hebei ranked at the top among all the provinces and iron & steel industry as the biggest source. Low-chlorinated PCNs comprised 90% of emissions by mass, while highly chlorinated PCNs dominated in terms of toxicity, highlighting divergent priorities for mitigating emissions and safeguarding human health. The emissions showed an overall upward trend from 1960 to 2019 driven by emission increase from iron & steel industry in terms of source, and from North China and East China in terms of geographic area. Per-capita emissions followed an inverted U-shaped environmental Kuznets curve while emission intensities decreased with increasing per-capita Gross Domestic Product (GDP) following a nearly linear pattern when log-transformed.

Ultrasonic humidifier aerosols: Observed high heavy metal enrichment and a new emission control method.

Ultrasonic humidifiers are commonly used in households to maintain indoor humidity and generate a large number of droplets or spray aerosols. However, there have been various health concerns associated with humidifier use, largely due to aerosols generated during operation. Here, we investigated the size distribution, chemical composition, and charged fraction of aerosol particles emitted from commercial ultrasonic humidifiers. Heavy metals in water used for humidifiers were found to be highly enriched in the ultrasonic humidifier aerosols (UHA), with the enrichment factors ranging from 102 to 107. This enrichment may pose health concerns for the building occupants, as UHA concentrations of up to 106 particles/cm3 or 3 mg/m3 were observed. Furthermore, approximately 90% of UHA were observed to be electrically charged, for the first time according to our knowledge. Based on this discovery, we proposed and tested a new method to remove UHA by using a simple electrical field. The designed electrical field in this work can efficiently remove 81.4% of UHA. Therefore, applying this electrical field could be an effective method to significantly reduce the health risks by UHA.

Small molecule induced interfacial defect healing to construct inverted perovskite solar cells with high fill factor and stability.

Chemical defects at the surface and grain boundaries of perovskite crystals cause deterioration of conversion efficiency and stability of perovskite solar cells (PSCs). In this study, a multifunctional additive, 5-fluoro-2-pyrimidine carbonitrile (FPDCN) molecule, is added into the perovskite precursor solution in order to passivate the uncoordinated Pb2+ by the cyanogen (-CN) group and pyrimidine N in FPDCN. Interestingly, fluorine (F) atoms interact with FA+ to form hydrogen bonds, which could regulate the perovskite crystallization process for the formation of high-quality perovskite crystals. Besides, the F atoms in FPDCN increase the water contact angle of perovskite films. As a result, the carrier extraction and transport in the perovskite film are significantly enhanced, and the non-radiative recombination is suppressed. The corresponding devices achieve a champion photovoltaic conversion efficiency (PCE) of 20.7 % and a fill factor (FF) of over 83 %. The device based on FPDCN shows long-term stability under a high-humidity atmospheric environment by maintaining 85 % of the initial efficiency after aging of 700 h in the glove box. This study provides a simple and convenient method to prepare stable and efficient PSCs by optimizing the perovskite precursor solution.

Metabolic signatures of metabolites of the purine degradation pathway in human plasma using HILIC UHPLC-HRMS.

The metabolic disorders in the purine degradation pathway have proven to be closely associated with several human diseases. However, the etiology is not yet fully understood. Profile assay of purine intermediates and uric acid involved in the metabolic pathway can provide additional insight into the nature and severity of related diseases. Purine metabolites are endogenous chemicals with high hydrophilicity, polarity, and similar structures, thus there is a great need for a specific method to quantify them directly in biological fluids with a short running time. Herein, eight purine degradation pathway metabolites, including xanthine, hypoxanthine, guanine, xanthosine, inosine, guanosine, adenosine and uric acid, in human plasma were quantitatively measured using hydrophilic interaction chromatography-tandem high-resolution mass spectrometry (HILIC-HRMS) in a short running time of 10 min. The method was systematically validated for specificity, linearity of the calibration curve, the limit of detection, the limit of quantification, the lower limit of quantification, precision, accuracy, extraction recovery, matrix effect, and stability. The results showed that the method was linear (R2 > 0.99), accurate (the intra- and inter-day recoveries of all analytes ranged from 90.0 % to 110.0 %), and precise (the intra- and inter-day precisions were less than 6.7 % and 8.9 %, respectively) with the lower limits of quantification ranging from 3 to 10,000 ng/mL. The extraction recoveries and matrix effects were repeatable and stable. All the analytes were stable in the autosampler and could be subject to three freeze-thaw cycles. The developed method was ultimately applied to 100 plasma specimens from healthy individuals. The results showed that the concentrations of different purine metabolites varied dramatically in plasma specimens. Diet and body mass index (BMI) were the most significant factors determining purine levels, followed by drinking and sex. Age, smoking and bedtime showed a very weak correlation with purine metabolism. The findings of the present work reveal the characteristics of purine metabolism in human plasma under non-pathological conditions. The results also highlight the factors that can cause changes in purine metabolism, which are useful in developing effective treatment strategies for metabolic disorders of purines, particularly for those caused by lifestyle factors.

Marine thermal fluctuation induced gluconeogenesis by the transcriptional regulation of CgCREBL2 in Pacific oysters.

Marine thermal fluctuation profoundly influences energy metabolism, physiology, and survival of marine life. In the present study, short-term and long-term high-temperature stresses were found to affect gluconeogenesis by inhibiting PEPCK activity in the Pacific oyster (Crassostrea gigas), which is a globally distributed species that encounters significant marine thermal fluctuations in intertidal zones worldwide. CgCREBL2, a key molecule in the regulation of gluconeogenesis, plays a critical role in the transcriptional regulation of PEPCK in gluconeogenesis against high-temperature stress. CgCREBL2 was able to increase the transcription of CgPEPCK by either binding the promoter of CgPEPCK gene or activating CgPGC-1α and CgHNF-4α after short-term (6 h) high-temperature stress, while only by binding CgPEPCK after long-term (60 h) high-temperature stress. These findings will further our understanding of the effect of marine thermal fluctuation on energy metabolism on marine organisms.

FGF20 modulates gut microbiota to mitigate dextran sodium sulfate-induced ulcerative colitis in mouse models.

Ulcerative colitis (UC), a prevalent form of inflammatory bowel disease (IBD), presents a significant clinical challenge due to the lack of optimal therapeutic strategies. Emerging evidence suggests that fibroblast growth factor 20 (FGF20) may play a crucial role in mitigating UC symptoms, though the mechanistic underpinnings remain elusive. In this study, a mouse model of UC was established using dextran sodium sulfate (DSS) to investigate the potential role of FGF20. Our findings revealed a marked reduction in FGF20 expression in the serum and colonic tissues of DSS-treated mice. Furthermore, FGF20 knockout did not exacerbate colonic damage in these mice. Conversely, overexpression of FGF20 via adeno-associated virus (AAV) significantly alleviated UC-associated symptoms. This alleviation was evidenced by attenuated intestinal shortening, mitigated weight loss, increased colonic goblet cell density and crypt formation, reduced inflammation severity and inflammatory cell infiltration, and enhanced expression of tight junction and mucin proteins. Moreover, FGF20 significantly ameliorated the dysbiosis of gut microbiota in DSS-treated mice by increasing the abundance of beneficial bacteria and decreasing the abundance of harmful bacteria. The beneficial effects of FGF20 were notably attenuated following gut microbiota depletion with an antibiotic regimen. Fecal microbiota transplantation experiments further supported the critical role of gut microbiota in mediating the effects of FGF20 on DSS-treated mice. In conclusion, these findings highlight the potential involvement of gut microbiota in the therapeutic effects of FGF20 in UC.

Meningeal lymphatic function promotes oligodendrocyte survival and brain myelination.

The precise neurophysiological changes prompted by meningeal lymphatic dysfunction remain unclear. Here, we showed that inducing meningeal lymphatic vessel ablation in adult mice led to gene expression changes in glial cells, followed by reductions in mature oligodendrocyte numbers and specific lipid species in the brain. These phenomena were accompanied by altered meningeal adaptive immunity and brain myeloid cell activation. During brain remyelination, meningeal lymphatic dysfunction provoked a state of immunosuppression in the brain that contributed to delayed spontaneous oligodendrocyte replenishment and axonal loss. The deficiencies in mature oligodendrocytes and neuroinflammation due to impaired meningeal lymphatic function were solely recapitulated in immunocompetent mice. Patients diagnosed with multiple sclerosis presented reduced vascular endothelial growth factor C in the cerebrospinal fluid, particularly shortly after clinical relapses, possibly indicative of poor meningeal lymphatic function. These data demonstrate that meningeal lymphatics regulate oligodendrocyte function and brain myelination, which might have implications for human demyelinating diseases.

The progress and promise of lineage reprogramming strategies for liver regeneration.

The liver exhibits remarkable regenerative capacity. However, the limited ability of primary human hepatocytes to proliferate in vitro, combined with a compromised regenerative capacity induced by pathological conditions in vivo, presents significant obstacles to effective liver regeneration following liver injuries and diseases. Developing strategies to compensate for the loss of endogenous hepatocytes is crucial for overcoming these challenges, and this remains an active area of investigation. Lineage reprogramming, the process of directly converting one cell type into another bypassing the intermediate pluripotent state, has emerged as a promising method for generating specific cell types for therapeutic purposes in regenerative medicine. Here, we discuss the recent progress and emergent technologies in lineage reprogramming into hepatic cells, and their potential applications in enhancing liver regeneration or treating liver disease models. We also address controversies and challenges that confront this field.

Effects of Platelet-Activating Factor (PAF) on the Mechanical Activities of Lower Urinary Tract and Genital Smooth Muscles.

Since its first discovery as a bioactive phospholipid inducing potent platelet aggregation, platelet-activating factor (PAF) has been shown to be involved in a wide variety of inflammatory and allergic disease states. Many pharmacological studies in the 1980s and 1990s also showed that PAF induces endothelium-dependent vascular relaxation and contraction of various smooth muscles (SMs), including those in the airway, gastrointestinal organs, and uterus. However, since the late 1990s, there have been few reports on the SM contractions induced by PAF. The lower urinary tract (LUT), particularly the urinary bladder (UB) has attracted recent attention in SM pharmacology research because patients with LUT dysfunctions including overactive bladder are increasing as the population ages. In addition, recent clinical studies have implicated the substantial role of PAF in the inflammatory state in LUT because its production increases with smoking and with cancer. However, the effects of PAF on mechanical activities of LUT SMs including UBSM have not been investigated to date. Recently, we found that PAF very strongly increased mechanical activities of UBSM in guinea pigs and mice, and partly elucidated the possible mechanisms underlying these actions of PAF. In this review, we describe the effects of PAF on LUT SMs by introducing our recent findings obtained in isolated UBSMs and discuss the physiological and pathophysiological significance. We also introduce our data showing the effects of PAF on the SM mechanical activities of genital tissues (prostate and vas deferens).

Collagen-binding bone morphogenetic protein-2 designed for use in bone tissue engineering.

Bone tissue engineering using biodegradable porous scaffolds is a promising approach for restoring oral and maxillofacial bone defects. Recently, attempts have been made to incorporate proteins such as growth factors to create bioactive scaffolds that can engage cells to promote tissue formation. Collagen-based scaffolds containing bone morphogenetic protein-2 (BMP2) have been studied for bone formation. However, controlling the initial burst of BMP2 remains difficult. Here we designed a functional chimeric protein composed of BMP2 and a collagen-binding domain (CBD), specifically the A3 domain of von Willebrand factor, to sustain BMP2 release from collagen-based scaffolds. Based on the results of computer-based structural prediction, we prepared a chimeric protein consisting of CBD and BMP2 in this order with a peptide tag for affinity purification. The chimeric protein had a collagen-binding capacity and enhanced osteogenic differentiation of human mesenchymal stem cells. These results are consistent with insights from in silico structural prediction.

Climate risk and green total factor productivity in agriculture: The moderating role of climate policy uncertainty.

In light of the escalating global warming and the escalating frequency of extreme weather events, the agricultural sector, being a fundamental and pivotal industry worldwide, is encountering substantial challenges due to climate change. Using Chinese provincial panel data for 2000-2021, this paper utilizes a two-way fixed-effect model to investigate the impact of Climate Risk (CR) on green total factor productivity in agriculture (AGTFP), with China's climate policy uncertainty (CPU) being introduced as a moderating variable within the research framework to scrutinize its influence in this context. The findings reveal a noteworthy adverse effect of CR on AGTFP, further exacerbated by CPU. Heterogeneity analysis results show that there is a clear regional variation in the effect of CR on AGTFP across different Chinese regions, with CR significantly inhibiting AGTFP development in the northern regions and provinces in major grain producing regions. Consequently, there is a pressing necessity to bolster the establishment of climate change monitoring infrastructures, devise tailored climate adaptation strategies at a regional level, and enhance the clarity and predictability of climate policies to fortify the resilience and sustainability of agricultural production systems.

Immune-mediated impairment of tonic immobility defensive behavior in an experimental model of colonic inflammation.

Ulcerative colitis has been associated with psychological distress and an aberrant immune response. The immunomodulatory role of systemic cytokines produced during experimental intestinal inflammation in tonic immobility (TI) defensive behavior remains unknown. The present study characterized the TI defensive behavior of guinea pigs subjected to colitis induction at the acute stage and after recovery from intestinal mucosa injury. Moreover, we investigated whether inflammatory mediators (tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-8, IL-10, and prostaglandins) act on the mesencephalic nucleus, periaqueductal gray matter (PAG). Colitis was induced in guinea pigs by intrarectal administration of acetic acid. The TI defensive behavior, histology, cytokine production, and expression of c-FOS, IBA-1, and cyclooxygenase (COX)-2 in PAG were evaluated. Colitis reduced the duration of TI episodes from the first day, persisting throughout the 7-day experimental period. Neuronal c-FOS immunoreactivity was augmented in both columns of the PAG (ventrolateral (vlPAG) and dorsal), but there were no changes in IBA-1 expression. Dexamethasone, infliximab, and parecoxib treatments increased the duration of TI episodes, suggesting a modulatory role of peripheral inflammatory mediators in this behavior. Immunoneutralization of TNF-α, IL-1ß, and IL-8 in the vlPAG reversed all effects produced by colitis. In contrast, IL-10 neutralization further reduced the duration of TI episodes. Our results reveal that peripherally produced inflammatory mediators during colitis may modulate neuronal functioning in mesencephalic structures such as vlPAG.

Effect of smart city construction on an urban transition to a low-carbon economy: evidence from China.

China is currently in a new era of an urban transition to a low-carbon economy and digital economic development. Smart cities, as an advanced form of information-based urban development, may be the key to the urban transition to low-carbon emissions. This paper examined the effect of smart city construction (SCC) on urban low-carbon transitions and its transmission mechanisms in China from the dual perspectives of reducing urban total carbon emissions (TCE) and improving urban total-factor carbon emission efficiency (TFCEE). Utilizing a multi-period difference in differences (DID) method, this study was conducted based on panel data of 245 Chinese prefecture-level cities from 2003 to 2021. The results demonstrated that SCC both reduced TCE and enhanced TFCEE. The effects of SCC were stronger in cities with more stringent environmental regulations. SCC achieved the dual effect of reducing TCE and enhancing urban TFCEE by promoting green technological progress and a low-carbon transformation of city residents' lifestyles. Moreover, optimization of the industrial structure was also a transmission mechanism for SCC to improve TFCEE. These conclusions provide an empirical basis for the SCC to empower low-carbon transitions of cities and help countries in different regions to transform the extensive urban development mode and promote urban low-carbon economic development.

miR­155 promotes an inflammatory response in HaCaT cells via the IRF2BP2/KLF2/NF­κB pathway in psoriasis.

Psoriasis is a chronic inflammatory skin condition with numerous causes, including genetic, immunological and infectious factors. The course of psoriasis is long and recurrence is common; pathogenesis is not completely understood. However, there is an association between advancement of psoriasis and aberrant microRNA (miR or miRNA)­155 expression. Through bioinformatics, the present study aimed to analyze the differentially expressed genes and miRNAs in psoriasis and its biological mechanism and function psoriatic inflammation. First of all, differentially expressed genes (DEGs) and miRNAs (DEMs) in patients with psoriasis were identified using GEO2R interactive web application. A psoriasis inflammatory model was established using lipopolysaccharide (LPS)­treated HaCaT keratinocytes, which were transfected with miR­155 mimic or inhibitor. Cell Counting Kit­8 was used for the assessment of cell viability and proliferation, and changes in the cell cycle were examined using flow cytometry. ELISA and reverse transcription­quantitative PCR (RT­qPCR) were used to detect the expression levels of the inflammatory factors IL­1ß and IL­6. The dual­luciferase reporter assay was used to verify the targeting association between miR­155­5p and IFN regulatory factor 2 binding protein 2 (IRF2BP2). To verify the targeting association of miR­155 and the IRF2BP2/kruppel­like factor 2 (KLF2)/NF­κB signaling pathway, expression levels of IRF2BP2, KLF2 and p65 were identified by RT­qPCR and western blotting. IRF2BP2 levels were also confirmed by immunofluorescence, in conjunction with bioinformatics database analysis. Overexpression of miR­155 inhibited proliferation of HaCaT cells and increased the number of cells in S phase and decreasing number of cells in G1 and G2 phase. In the LPS­induced inflammatory state, miR­155 overexpression heightened the inflammatory response of HaCaT cells while inhibition of miR­155 lessened it. Suppression of inflammatory cytokine expression by miR­155­5p inhibitor was reversed by knockdown of IRF2BP2. miR­155 was shown to interact with IRF2BP2 to negatively regulate its expression, leading to decreased KLF2 expression and increased p65 expression and secretion of inflammatory factors, intensifying the inflammatory response of HaCaT cells. Therefore, miR­155 may contribute to development of psoriasis by inducing tissue and cell damage by increasing the inflammatory response of HaCaT cells via the IRF2BP2/KLF2/NF­κB pathway. In conclusion, the results of the present study offer novel perspectives on the role of miR­155 in the onset and progression of psoriasis.

Prognostic value of neutrophil to lymphocyte ratio in patients with advanced pancreatic ductal adenocarcinoma treated with systemic chemotherapy.

OBJECTIVES: Although systemic chemotherapy for pancreatic ductal adenocarcinoma (PDAC) has made progress, ensuring long-term survival remains difficult. There are several reports on the usefulness of neutrophil-to-lymphocyte ratio (NLR) in predicting the prognosis of PDAC, but few reports in systemic chemotherapy. We hereby investigated the usefulness of NLR in systemic chemotherapy for PDAC. MATERIALS AND METHODS: A retrospective study was conducted on patients with advanced PDAC treated with first-line systemic chemotherapy. Cox regression hazards models were performed to analyze the association between baseline patient characteristics and the initial treatment response, and overall survival (OS). RESULTS: A total of 60 patients with PDAC were enrolled. At baseline, there were significant differences in NLR and carbohydrate antigen 19-9 (CA19-9), as well as the selection rate of combination chemotherapy, between patients with partial response or stable disease and those with progressive disease. Univariate and multivariate analysis showed that NLR < 3.10, combination chemotherapy, and CA19-9 < 1011 U/mL were significant and independent predictive factors of the initial treatment response. Meanwhile, NLR < 3.10 and combination chemotherapy were independently associated with longer OS. Moreover, OS was significantly prolonged in patients with NLR < 3.10, regardless of whether combination chemotherapy or monotherapy. Patients with NLR < 3.10 at baseline had a significantly higher conversion rate to third-line chemotherapy and a longer duration of total chemotherapy. CONCLUSIONS: This study suggests that NLR may be a useful marker for predicting the initial treatment response to first-line chemotherapy and the prognosis for patients with advanced PDAC.

Three-dimensional spectrochromatographic determination of chlorogenic acid in Melampyrum stenophyllum Boiss. extracts by parallel factor analysis.

INTRODUCTION: Co-elution is a common challenge in phytochemical chromatography. Full chromatographic separation often requires extensive optimization, long analysis times, and excessive solvent use. A viable alternative could be mathematical elution of analytes using three-dimensional decomposition. OBJECTIVES: This study aimed to develop a method to determine chlorogenic acid in Melampyrum stenophyllum Boiss. extracts without complete chromatographic separation, to validate the method, and to cross-validate assay results against a classical ultra-performance liquid chromatography (UPLC) method. METHODOLOGY: Ultra-performance liquid chromatography-photodiode array (UPLC-PDA) spectrochromatograms were arranged into a three-way data cube with dimensions of time, wavelength, and sample and then decomposed using parallel factor analysis to reveal chromatographic, spectral, and concentration profiles. The chromatographic and spectral profiles were used to identify chlorogenic acid in overlapping signals. The relative concentration profile was used to quantify it in the plant extract. The assay results were statistically compared with those from an in-house classical UPLC method. RESULTS: Chlorogenic acid was co-eluted at 1.45 min and quantified as 16.11 mg per gram dry weight of Melampyrum stenophyllum extracts (SD = 0.28), despite significant interference in a 4-min runtime. The analytical validity was confirmed by recovery calculations from standard solutions and standard addition samples (RSD < 2%), and the t-test resulted in a p-value of 0.09 (α = 0.05), indicating no significant difference between the results obtained from mathematical elution and chromatographic separation. CONCLUSION: Chlorogenic acid was quantified from plant material accurately despite the co-elution. Validation and cross-validation results support the method's applicability.

Synergistic Interaction Between PbbZIP88 and PbSRK2E Enhances Drought Resistance in Pear Through Regulation of PbATL18 Expression and Stomatal Closure.

Drought poses significant challenges to agricultural production, ecological stability and global food security. While wild pear trees exhibit strong drought resistance, cultivated varieties show weaker drought tolerance. This study aims to elucidate the molecular mechanisms underlying pear trees' response to drought stress. We identified a drought resistance-related transcription factor, PbbZIP88, which binds to and activates the expression of the drought-responsive gene PbATL18. Overexpression of PbbZIP88 in Arabidopsis and pear seedlings resulted in enhanced drought resistance and significantly improved physiological parameters under drought stress. We discovered that PbbZIP88 interacts with the key protein PbSRK2E in the ABA signalling pathway. This interaction enhances PbbZIP88's ability to activate PbATL18 expression, leading to higher levels of PbATL18. Furthermore, the PbbZIP88 and PbSRK2E interaction accelerates the regulation of stomatal closure under ABA treatment conditions, reducing water loss more effectively. Experimental evidence showed that silencing PbbZIP88 and PbSRK2E genes significantly decreased drought resistance in pear seedlings. In conclusion, this study reveals the synergistic role of PbbZIP88 and PbSRK2E in enhancing drought resistance in pear trees, particularly in the upregulation of PbATL18 expression, and the accelerated promotion of stomatal closure. These findings provide new candidate genes for breeding drought-resistant varieties and offer a theoretical foundation and technical support for achieving sustainable agriculture.

Empathy ability and influencing factors among pediatric residents in China: a mixed-methods study.

BACKGROUND: Empathy is one of the fundamental factors enhancing the therapeutic effects of physician-patient relationships, but there has been no relevant research in China on the pediatric resident physicians' capacity for empathy or the influencing factors. METHODS: A mixed-methods study was undertaken. The student version of the Jefferson Scale of Empathy was used to assess 181 postgraduate residents at Shanghai Children's Medical Center and Shanghai Children's Hospital. Differences in empathy ability among pediatric resident physicians of different genders and specialties were analyzed using independent sample t-tests and Mann-Whitney U tests. A one-way analysis of variance was used to analyze the differences in empathy ability at different educational levels and years of medical residency training. Seven third-year postgraduate pediatric residents from Shanghai Children's Medical Center participated in semi-structured interviews exploring the influencing factors. We analyzed the interview transcripts using thematic analysis. RESULTS: The scale was completed by 154 pediatric residents. No statistically significant differences in empathy were found between educational level, postgraduate year, gender, or specialty. The factors influencing empathy in doctor-patient communication included the person who accompanied the child to see the doctor, how the children cooperated with doctors for medical treatment, the volume of pediatric outpatient and emergency visits, and the physician's ability to withstand pressure. All interviewed resident physicians regarded learning empathy as important but rarely spent extra time learning it. CONCLUSIONS: The evaluation results of resident physicians on changes in empathy after improving clinical abilities vary according to their understanding of empathy, and the work environment has an important impact on pediatricians' empathy ability. Their empathy score is relatively low, and this requires exploration and intervention.