A new method of alkalinity remediation for Cd-contaminated groundwater by PAAS-modified MgCO3/Mg(OH)2 colloid.

Mg(OH)2 dissolves slowly and can provide a long-term source of alkalinity, thus a promising alternative reagent for the in situ remediation of heavy metal polluted groundwater. Unfortunately, it exhibits a relatively poor stabilization effect on heavy metal Cd due to the higher solubility of the resulting stabilized product, Cd(OH)2. To overcome this limitation, we investigated the use of MgCO3/Mg(OH)2 colloid modified by sodium polyacrylate (PAAS) to remove Cd from groundwater. Through ultrasonic dispersion, the molecular chains of PAAS are broken, causing a transformation from flocculation to surface modification, resulting in the production of a stable colloid. The colloidal particles of MgCO3/Mg(OH)2 have a smaller size and a negatively charged surface, which significantly enhances their migration ability in aquifers. The combination of MgCO3 and Mg(OH)2 provides a complementary effect, where MgCO3 effectively precipitates Cd in the aquifer while Mg(OH)2 maintains the required pH level for stabilization. The optimal compounding ratio of MgCO3 to Mg(OH)2 for achieving the best stabilization effect on Cd is found to be 1:1. Column experiments demonstrate that the injection of MgCO3/Mg(OH)2 colloid substantially enhances Cd stability, reducing the exchangeable fraction of Cd in aquifer media from 88.61% to a range of 22.50-34.38%. Based on these results, the MgCO3/Mg(OH)2 colloid shows great potential as a reactive medium for remediating Cd-contaminated groundwater.

Site-Directed Mutagenesis of VvCYP76F14 (Cytochrome P450) Unveils Its Potential for Selection in Wine Grape Varieties Linked to the Development of Wine Bouquet.

Bouquet is a fascinating wine characteristic that serves as an indicator of wine quality, developing during the aging process. The multifunctional monoterpenol oxidase VvCYP76F14 in wine grapes sequentially catalyzes three reactions to produce (E)-8-carboxylinalool, a crucial precursor for wine bouquet. Previous studies indicated that the activity of VvCYP76F14 derived from different wine grape varieties did not correlate with the amino acid sequence differences. In this study, 54 wine grape varieties were categorized into neutral, aromatic, and full-bodied types based on the sequence differences of VvCYP76F14, closely correlated with the content of wine lactone precursors. Computer modeling and molecular docking analysis of the full-bodied CYP76F14 revealed 17, 19, and 18 amino acid residues in the VvCYP76F14-linalool, VvCYP76F14-(E)-8-hydroxylinalool, and VvCYP76F14-(E)-8-oxolinalool complexes, respectively. Site-directed mutagenesis and in vitro enzyme activity analysis confirmed the substitutions of the key amino acid residues in neutral and aromatic varieties. Notably, the D299 mutation of VvCYP76F14 resulted in the complete loss of (E)-8-oxolinalool and (E)-8-carboxylinalool activities, aligning with the undetectable levels of (E)-8-oxolinalool and (E)-8-carboxylinalool in "Yantai 2-3-37", which harbors the D299T substitution. Favorably, VvCYP76F14 could serve as a cost-effective fingerprint marker for screening superior hybrid offspring with the desired levels of wine lactone precursors.

The haplotype-resolved T2T genome of teinturier cultivar Yan73 reveals the genetic basis of anthocyanin biosynthesis in grapes.

Teinturier grapes are characterized by the typical accumulation of anthocyanins in grape skin, flesh, and vegetative tissues, endowing them with high utility value in red wine blending and nutrient-enriched foods developing. However, due to the lack of genome information, the mechanism involved in regulating teinturier grape coloring has not yet been elucidated and their genetic utilization research is still insufficient. Here, the cultivar 'Yan73' was used for assembling the telomere-to-telomere (T2T) genome of teinturier grapes by combining the High Fidelity (HiFi)， Hi-C and ultralong Oxford Nanopore Technologies (ONT) reads. Two haplotype genomes were assembled, at the sizes of 501.68 Mb and 493.38 Mb, respectively. In the haplotype 1 genome, the transposable elements (TEs) contained 32.77% of long terminal repeats (LTRs), while in the haplotype 2 genome, 31.53% of LTRs were detected in TEs. Furthermore, obvious inversions were identified in chromosome 18 between the two haplotypes. Transcriptome profiling suggested that the gene expression patterns in 'Cabernet Sauvignon' and 'Yan73' were diverse depending on tissues, developmental stages, and varieties. The transcription program of genes in the anthocyanins biosynthesis pathway between the two cultivars exhibited high similarity in different tissues and developmental stages, whereas the expression levels of numerous genes showed significant differences. Compared with other genes, the expression levels of VvMYBA1 and VvUFGT4 in all samples, VvCHS2 except in young shoots and VvPAL9 except in the E-L23 stage of 'Yan73' were higher than those of 'Cabernet Sauvignon'. Further sequence alignments revealed potential variant gene loci and structure variations of anthocyanins biosynthesis related genes and a 816 bp sequence insertion was found in the promoter of VvMYBA1 of 'Yan73' haplotype 2 genome. The 'Yan73' T2T genome assembly and comparative analysis provided valuable foundations for further revealing the coloring mechanism of teinturier grapes and the genetic improvement of grape coloring traits.

A prognostic nomogram and risk classification system of elderly patients with extraosseous plasmacytoma: a SEER database analysis.

BACKGROUND: The survival trends and prognostic factors of patients with extraosseous plasmacytoma (EOP) or extramedullary plasmacytoma (EMP) have not been reported in recent years. The objective of this study was to develop a novel nomogram and risk stratification system for predicting the overall survival (OS) of elderly patients with EOP based on the Surveillance, Epidemiology, and End Results (SEER) database. METHODS: The demographic characteristics of 900 patients aged 60 years and above, diagnosed with EOP between 2000 and 2019, were extracted from the SEER database. The patient population was randomly divided into a training cohort and an internal validation cohort in a ratio of 7:3. Univariate and multivariate Cox regression analyses were conducted to identify independent predictors of prognosis in elderly EOP patients, followed by developing a nomogram for prognostic assessment. The performance of the model was evaluated through receiver-operating characteristic (ROC) curves, C-index, calibration curves for calibration accuracy assessment, and decision curve analysis (DCA) to assess its clinical utility. All elderly EOP patients were stratified into three risk subgroups by cutoff value utilizing X-tile software based on their total OS scores for comparative analysis purposes. Kaplan-Meier (K-M) survival curve analysis was employed to validate any observed differences in OS among these three risk groups. RESULTS: Six factors including age, year of diagnosis, marital status, primary site, surgery, and prior tumor history were identified to be independently predictive of the OS of elderly patients with EOP, and these predictors were included in the construction of the nomogram. The 1-, 3-, and 5-year area under the curves (AUCs) for OS were 0.717, 0.754, and 0.734 in the training cohort and 0.740, 0.730, and 0.765 in the validation cohort, respectively. The C-index values in the two cohorts were 0.695 and 0.690. The calibration curves and DCA exhibit commendable consistency and validity, respectively, thereby demonstrating their robust performance. The training set was stratified into low-, medium-, and high-risk subgroups based on the optimal cutoff points (167.8 and 264.8) identified. The K-M curve and cumulative risk curve exhibited statistically significant disparities in survival rates among the groups. CONCLUSIONS: We developed a nomogram and risk classification system, which can serve as an intuitive and effective tool for clinicians to enhance the prediction of OS in elderly EOP patients, thereby facilitating the formulation of more rational and personalized treatment strategies.

Association between urine caffeine metabolites and bone mineral density: A population-based study.

The association between coffee intake and bone mineral density (BMD) remains a subject of debate in epidemiological research. Furthermore, the potential relationship between BMD and urine caffeine or caffeine metabolites has not yet been explored. Therefore, the present study aimed to investigate the possible association between BMD and urine caffeine and its metabolites in U.S. adults. We employed multivariate linear and logistic regression models to analyze the relationship between urine caffeine and caffeine metabolites and lumbar BMD using data from the National Health and Nutrition Examination Survey (NHANES) from 2009 to 2014. Additionally, fitted smoothing curves and generalized additive models were used. After adjusting for several factors, we found no significant association between urine caffeine and its metabolites and BMD. However, subgroup analyses stratified by gender and ethnicity showed that the relationship between urine caffeine and its metabolites and lumbar BMD remained consistent. Our investigation revealed that the inflection points for the U-shaped relationship between urinary theophylline and paraxanthine and BMD were observed at levels of 0.006 mmol/L for theophylline and 0.052 mmol/L for paraxanthine. In this cross-sectional study, we found no significant correlation between urine caffeine and its metabolites and BMD. However, more research is required to confirm our findings, as well as to investigate the underlying mechanisms.

Association of family income to poverty ratio and vibration-controlled transient elastography quantified degree of hepatic steatosis in U.S. adolescents.

Introduction: Inequality in socioeconomic status plays an important role in the prevalence of metabolic diseases in adolescents. The purpose of this study was to explore the association between family income and the degree of hepatic steatosis quantified by vibration-controlled transient elastography (VCTE) among U.S. adolescents. Methods: This cross-sectional study included two cycles of the National Health and Nutrition Examination Survey (NHANES) 2017-2020. Multivariate linear regression and smoothing curve fitting were used to investigate the linear and nonlinear relationship between PIR and hepatic steatosis, respectively. Subgroup analysis and interaction tests were used to test whether this relationship was stable across groups. Results: Of the 1,574 adolescent participants, 456 lived in poor households and 307 lived in wealthy households. After adjusting for all covariates, PIR (Ratio of family income to poverty) was significantly negatively associated with the degree of hepatic steatosis [-4.78 (-7.39, -2.17)], and this remained stable after converting PIR to a categorical variable. In addition, this significant negative association was more pronounced in women [-7.62 (-11.38, -3.87)], non-Hispanic blacks [-7.19 (-14.43, 0.06)], Mexican Americans [-6.80 (-13.63, 0.03)], and participants with BMI >30 cm2 [-10.83 (-19.70, -1.96)]. Conclusions: PIR was significantly and negatively associated with the degree of hepatic steatosis in US adolescents. Additional prospective studies are needed to confirm our findings.

Global trends in research on oxidative stress related to heart failure from 2012 to 2021: a bibliometric analysis and suggestion to researchers.

Background: Oxidative stress leads to an increase in reactive oxygen in the body. During heart failure (HF), when the body's antioxidant defense system fails to remove excessive reactive oxygen species, myocardial cells will be damaged or even die. Over the past ten years, the number of research publications on oxidative stress related to HF has increased. Methods: We searched publications published in 2012-2021 and the Web of Science Core Collection (WoSCC) recording information. Based on the VOSviewer and CiteSpace, we conducted a bibliometric analysis of the overall distribution of journals, keywords, authors, major countries, annual output, active institutions, and cocited literature. The Global Citation Score (GCS) was used to evaluate the impact and quality of highly cited papers. Results: We retrieved 5,616 articles and reviews. Over the past ten years, the number of annual publications on oxidative stress related to HF has increased. USA has published the largest number of articles and obtained the highest number of citations (NC) and H-index. The University of California and PLoS One are the most productive affiliations and journals in terms of publications on oxidative stress related to HF. The GCS of articles written by Paulus WJ in 2013 was 1,632, which was the top ranking. The most frequent keywords are "oxidative stress", "heart failure", "inflammation", "dysfunction" and "apoptosis". The top three authors are Kang Yuming, Ren Jun and Okoshi Katashi. "Impact", "induced myocardial infarction", "cardiovascular outcome", "empagliflozin", "sglt2 inhibitor", "protect", and "Na+/H+ exchanger" have become popular research topics. Conclusions: Our research shows the research focus and development trends of oxidative stress related to HF in the past decade. Understanding the most important indicators of oxidative stress related to HF and the hot spots in the field of oxidative stress research related to HF can assist scholars, countries and policy-makers in the field in better understanding oxidative stress related to HF and can also lead to better decisions in oxidative stress treatment.

Clinical features of infants with SARS-CoV-2 infection: a systematic review and meta-analysis.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) leads to coronavirus disease 2019 (COVID-19) and is a public health problem. This meta-analysis reviewed the clinical features of SARS-CoV-2 infection among infants. METHODS: PubMed, Scopus, Web of Science, and the Cochrane Library were searched for studies on clinical features of infants with SARS-CoV-2 published before May 1, 2022. Two authors screened and extracted data on the number of infants with SARS-CoV-2 infection, clinical features, and number of clinical features. The proportion of asymptomatic infection, mild symptoms, moderate symptoms, severe symptoms, and the clinical features were analyzed. RESULTS: Forty-four studies with 6,304 infants with SARS-CoV-2 infections were included in this study. The proportion of asymptomatic infection was 20% (95% CI: 11-28%, I2=97%, P<0.01) in infants with SARS-CoV-2 infections. The proportion of infants with mild, moderate, and severe symptoms was 48% (95% CI: 30-65%, I2=96%, P<0.01), 27% (95% CI: 10-44%, I2=93%, P<0.01), and 8% (95% CI: 0-16%, I2=90%, P<0.01), respectively. Notably, the most common clinical features of infants with SARS-CoV-2 infection were fever (64%), cough (34%), and nasal symptoms (31%). CONCLUSIONS: This meta-analysis found that 20% of infants with SARS-CoV-2 infections were asymptomatic, while most infants with COVID-19 presented with mild symptoms.

Diagnostic utility of rapid sequencing in critically ill infants: a systematic review and meta-analysis.

BACKGROUND: Genetic disorders are a major cause of death in critically ill infants. Several studies have assessed the diagnostic yield of rapid genomic sequencing in critically ill infants. This meta-analysis aimed to summarize the diagnostic utility of rapid genomic sequencing in critically ill infants. METHODS: PubMed, Scopus, Web of Science, and Cochrane Library, were searched before 1 July 2022. Studies reported diagnostic rate of rapid genomic sequencing in critically ill infants were selected. Two authors screened and extracted data regarding the method of genetic test, total number of patients, and number of diagnosed patients. RESULTS: Twenty-three studies, comprising 1567 critically ill infants were included in the meta-analysis. In the overall analysis, the pooled diagnostic utility of rapid genomic sequencing was 0.42 (95% CI: 0.37-0.49, I2 = 79%, P < 0.1). Moreover, the pooled diagnostic rates of rapid whole-exome and rapid whole-genome sequencing were 0.50 (95% CI: 0.41-0.61; I2 = 74%; P < 0.01) and 0.37 (95% CI: 0.30-0.46; I2 = 77%; P < 0.01), respectively. Sensitive analysis showed that the results were stable in the overall analysis. Additionally, publication bias was not observed in the overall analysis. CONCLUSIONS: This meta-analysis proved that rapid genomic sequencing has a good diagnostic utility for critically ill infants.

Estimation of hereditary fructose intolerance prevalence in the Chinese population.

BACKGROUND: Hereditary fructose intolerance (HFI) caused by aldolase B reduction or deficiency that results in fructose metabolism disorder. The disease prevalence in the Chinese population is unknown, which impedes the formulation of HFI screening and diagnosis strategies. MATERIALS AND METHODS: By searching a local cohort (Chinese Children's Rare Disease Genetic Testing Clinical Collaboration System, CCGT) and public databases (ClinVar and Human Gene Mutation Database) and reviewing HFI-related literature, we manually curated ALDOB pathogenic or likely pathogenic (P/LP) variants according to ACMG guidelines. Allele frequency (AF) information from the local database CCGT and the public databases HuaBiao and gnomAD for ALDOB P/LP variants was used to estimate and the HFI prevalence in the Chinese population and other populations by the Bayesian framework. We collected the genotype and clinical characteristics of HFI patients from the CCGT database and published literature to study genotype-phenotype relationships. RESULT: In total, 81 variants of ALDOB were curated as P/LP. The estimated Chinese HFI prevalence was approximately 1/504,678, which was much lower than that for non-Finland European (1/23,147), Finnish in Finland (1/55,539), admixed American (1/132,801) and Ashkenazi Jewish (1/263,150) populations. By analyzing the genetic characteristics of ALDOB in the Chinese population, two variants (A338V, A338G) had significantly higher AFs in the Chinese population than in the non-Finland European population from gnomAD (all P values < 0.05). Five variants (A150P, A175D, N335K, R60*, R304Q) had significantly lower AFs (all P values < 0.1). The genotype-phenotype association analyses were based on 68 reported HFI patients from a literature review and the CCGT database. The results showed that patients carrying homozygous variant sites (especially A150P) were more likely to present nausea, and patients carrying two missense variant sites were more likely to present aversion to sweets and fruit (all P values < 0.05). Our research reveals that some gastrointestinal symptoms seem to be associated with certain genotypes. CONCLUSION: The prevalence of HFI in the Chinese population is extremely low, and there is no need to add HFI testing to the current newborn screening programs if medical costs are considered. A genetic testing strategy is suggested for early diagnosis of HFI.

Application Effect of Whole-Process Seamless Nursing Model Based on Smart Healthcare Mode in Perioperative Period of Patients Undergoing Hematoma Removal.

OBJECTIVE: To explore the application effect of a whole-process seamless nursing model based on the smart healthcare mode in the perioperative period of patients undergoing hematoma removal. METHODS: In this retrospective study, 50 patients with hematoma removal admitted to our hospital from August 2018 to August 2019 were included as the control group (CG), while 50 patients with hematoma removal admitted to our hospital from September 2019 to September 2020 were included as the experimental group (EG). During the period of hematoma removal, CG received routine perioperative nursing, while EG received the whole-process seamless nursing model based on the smart healthcare mode. The perioperative indexes, hemodynamic indexes, and the incidence of postoperative complications were compared between the two groups, and the incidence of nursing staff's work omissions in different periods was analyzed. RESULTS: Notable differences were observed in surgical time, intraoperative blood loss, hematoma clearance rates, length of ICU stay, hospitalization time, removal time of ventricular drainage tube, and cerebral edema volume at 1 week after surgery between EG and CG (P < 0.05). Compared with CG, EG achieved obviously better hemodynamic indexes (P < 0.001) and a lower incidence of bedsore, muscle atrophy, and eating/swallowing disorders (P < 0.05). During the implementation of smart healthcare, the incidence of nursing staff's work omissions was remarkably reduced (P < 0.05). CONCLUSION: Under the smart healthcare, the incidence of nursing staff's work omissions is lower, and the effect of the whole-process seamless nursing is better, which can optimize the perioperative indexes of patients, stabilize the postoperative hemodynamics, and reduce the incidence of complications. Therefore, the whole-process seamless nursing model based on the smart healthcare mode has promotion value in clinic.

Paeoniflorin ameliorates chronic stress-induced depression-like behavior in mice model by affecting ERK1/2 pathway.

Depression is a mental and emotional disorder that has made an opening great burden to the society. Paeoniflorin showed remarkable antidepressant-like effects in multiple animal models with depressive disorders. However, the molecule of paeoniflorin on depression is less studied. This study aims to explore the effect and the molecular mechanism of paeoniflorin on depression in a chronic restraint stress (CRS) mice model. CRS model of C57BL/6 J mice was set up. Sucrose preference test (SPT), tail suspension test (TST), open field test (OFT) and forced swimming test (FST) were used to assess depression symptoms. Immunofluorescence staining, quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting were implemented to detect the expression changes of the proteins involved in extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway. Results showed that paeoniflorin treatment decreased the degree of depression in the CRS mice. Further analysis showed that the expression of ERK1/2 proteins was significantly downregulated, while paeoniflorin could elevate the expression of ERK1/2 proteins in CRS mice. Finally, it showed that inhibiting signaling ERK1/2 pathway could aggravate the depressive behavior when treatment with ERK-specific inhibitor U0126, while the condition could be partially relieved when treated with paeoniflorin. In conclusion, the present study demonstrated that paeoniflorin attenuated chronic stress-induced depression-like behavior in mice by affecting the ERK1/2 pathway. These findings provided the basis for the molecular mechanism of paeoniflorin on the effect of depression, which support paeoniflorin might act as an important drug in the treatment of depression.

HIGD­1B inhibits hypoxia­induced mitochondrial fragmentation by regulating OPA1 cleavage in cardiomyocytes.

The dynamic regulation of mitochondrial morphology is key for eukaryotic cells to manage physiological challenges. Therefore, it is important to understand the molecular basis of mitochondrial dynamic regulation. The aim of the present study was to explore the role of HIG1 hypoxia inducible domain family member 1B (HIGD­1B) in hypoxia­induced mitochondrial fragmentation. Protein expression was determined via western blotting. Immunofluorescence assays were performed to detect the subcellular location of HIGD­1B. Cell Counting Kit­8 assays and flow cytometry were carried out to measure cell viability and apoptosis, respectively. Protein interactions were evaluated by co­immunoprecipitation. In the present study, it was found that HIGD­1B serves a role in cell survival by maintaining the integrity of the mitochondria under hypoxic conditions. Knockdown of HIGD­1B promoted mitochondrial fragmentation, while overexpression of HIGD­1B increased survival by preventing activation of caspase­3 and ­9. HIGD­1B expression was associated with cell viability and apoptosis in cardiomyocytes. Furthermore, HIGD­1B delayed the cleavage process of optic atrophy 1 (OPA1) and stabilized mitochondrial morphology by interacting with OPA1. Collectively, the results from the present study identified a role for HIGD­1B as an inhibitor of the mitochondrial fission in cardiomyocytes.

Site-directed mutagenesis identified the key active site residues of alcohol acyltransferase PpAAT1 responsible for aroma biosynthesis in peach fruits.

The aroma of peach fruit is predominantly determined by the accumulation of γ-decalactone and ester compounds. A previous study showed that the biosynthesis of these aroma compounds in peach fruit is catalyzed by PpAAT1, an alcohol acyltransferase. In this work, we investigated the key active site residues responsible for γ-decalactone and ester biosynthesis. A total of 14 candidate amino acid residues possibly involved in internal esterification and 9 candidate amino acid residues possibly involved in esterification of PpAAT1 were assessed via site-directed mutagenesis. Analyses of the in vitro enzyme activities of PpAAT1 and its site-directed mutant proteins (PpAAT1-SMs) with different amino acid residue mutations as well as the contents of γ-decalactone in transgenic tobacco leaves and peach fruits transiently expressing PpAAT1 and PpAAT1-SMs revealed that site-directed mutation of H165 in the conserved HxxxD motif led to lost enzymatic activity of PpAAT1 in both internal esterification and its reactions, whereas mutation of the key amino acid residue D376 led to the total loss of γ-decalactone biosynthesis activity of PpAAT1. Mutations of 9 and 7 other amino acid residues also dramatically affected the enzymatic activity of PpAAT1 in the internal esterification and esterification reactions, respectively. Our findings provide a biochemical foundation for the mechanical biosynthesis of γ-decalactone and ester compounds catalyzed by PpAAT1 in peach fruits, which could be used to guide the molecular breeding of new peach species with more favorable aromas for consumers.

The effects of umbilical cord-derived macrophage exosomes loaded with cisplatin on the growth and drug resistance of ovarian cancer cells.

Objective: To assess the feasibility of an exosome-based drug delivery platform for the potent chemotherapeutic agent cisplatin to treat ovarian cancer.Significance: Exosomes have recently been used as drug delivery vehicles because of their natural advantages. Platinum-resistant forms of ovarian cancer require novel drug delivery methods to improve patient outcomes.Methods: We developed and compared different methods of loading exosomes released by mononuclear M1 and M2 macrophages from umbilical cord blood with cisplatin. We characterized the morphology, drug capacity, method of cellular entry, and antitumor efficacy of the exosomes in vitro.Results: Disruption of the exosomal membrane by sonication facilitated a high loading efficiency. Importantly, incorporation of cisplatin into umbilical cord blood-derived M1 macrophage exosomes increased its cytotoxicity 3.3× in drug-resistant A2780/DDP cells and 1.4× in drug-sensitive A2780 cells over chemotherapy alone. Loading of cisplatin into M2 exosomes increased its cytotoxicity by nearly 1.7× in drug-resistant A2780/DDP cells and 1.4× in drug-sensitive A2780 cells.Conclusions: We conclude that cisplatin-loaded M1 exosomes are potentially powerful new tools for the delivery of chemotherapeutics to treat cancers regardless of drug resistance.

Foliar Spraying with Compound Amino Acid-Iron Fertilizer Increases Leaf Fresh Weight, Photosynthesis, and Fe-S Cluster Gene Expression in Peach (Prunus persica (L.) Batsch).

As one of the most important micronutrients, iron (Fe) plays a critical role in various metabolic processes during plant growth and development. However, the molecular mechanisms towards Fe metabolism and nutrition in fruit trees are largely unknown. In this study, we examined the effects of amino acid-Fe compound fertilizer spraying on leaf development in peach (Prunus persica (L.) Batsch) at different developmental stages. Foliar spraying with amino acid-Fe compound fertilizer did not cause any significant changes in leaf morphology but remarkably increased leaf fresh weights. Fe concentration, photosynthetic parameter, and Fe-S protein analyses revealed that Fe accumulation, total chlorophyll content, net photosynthetic rate (P N), and stomatal conductance (g s), as well as nitrite reductase (NIR) and succinate dehydrogenase (SDH) activities, were significantly higher in the leaves sprayed with amino acid-Fe compound fertilizer than in the control leaves sprayed with distilled water. Further quantitative real-time PCR (qRT-PCR) analyses demonstrated that Fe-S cluster biosynthesis genes were differentially expressed in the leaves at different developmental stages. Foliar spraying with amino acid-Fe compound fertilizer significantly increased the expression of the most tested Fe-S cluster biosynthesis genes. Our findings provide new insights into the understanding of effects of Fe fertilization application on leaf development in perennial woody fruit trees.

Exosomes released from M2 macrophages transfer miR-221-3p contributed to EOC progression through targeting CDKN1B.

In contrast to other solid tumors within the abdominal cavity, epithelial ovarian cancers (EOCs) tend to undergo peritoneal metastasis. Thus, the peritoneal immune microenvironment is crucial for EOC progression. Previous reports indicate that the main immune cells within the peritoneum are M2 macrophages, specifically tumor-associated macrophages (TAMs). The communication between TAMs and tumor cells plays an important role in EOC development, and exosomes, acting as micro-message carriers, occupy an essential position in this process. Microarray analyses of exosomes revealed that miR-221-3p was enriched in M2 exosomes. Furthermore, miR-221-3p suppressed cyclin-dependent kinase inhibitor 1B (CDKN1B) directly. Thus, miR-221-3p contributed to the proliferation and G1/S transition of EOC cells. Additionally, low levels of CDKN1B were associated with EOC progression and poor prognosis. These observations suggest that TAMs-derived exosomal miR-221-3p acts as a regulator of EOC progression by targeting CDKN1B. The results of this study confirm that certain exosomal microRNAs may provide novel diagnostic biomarkers and therapeutic targets for EOC.

The exosomal integrin α5ß1/AEP complex derived from epithelial ovarian cancer cells promotes peritoneal metastasis through regulating mesothelial cell proliferation and migration.

PURPOSE: Epithelial ovarian cancer (EOC) is one of the most malignant cancers in the gynecologic system. Many patients are diagnosed at an advanced stage with disseminated intra-peritoneal metastases. EOC spreads via both direct extension and trans-coelomic spread. However, the interplay between human peritoneal mesothelial cells (HPMCs) and EOC cells is still ambiguous. We hypothesize that integrins (ITG) in HPMCs may play important roles in EOC metastasis. METHODS: The expression of different integrin subtypes from HPMCs was assessed using Western blotting. The expression of integrin α5ß1 (ITGA5B1) and its co-localization with asparaginyl endopeptidase (AEP) in HPMCs derived from EOC patients (EOC-HPMCs) were assessed using immunofluorescence. The role and mechanism of the exosomal ITGA5B1/AEP complex in HPMCs was assessed using both in vitro and in vivo assays. A retrospective study involving 234 cases was carried out to assess ITGA5B1 and AEP levels in circulating sera and ascites of EOC patients, as well as associations between ITGA5B1/AEP expression and overall survival. RESULTS: We found that ITGA5B1was highly expressed and co-localized with AEP in EOC cells, and that the exosomal ITGA5B1/AEP complex secreted by EOC cells played an important role in the proliferation and migration of HPMCs. High levels of exosomal ITGA5B1/AEP were also found in circulating sera and ascites of EOC patients, and the expression of ITGA5B1/AEP in EOC tissues was found to be negatively associated with overall survival. CONCLUSIONS: Our data indicate that EOCs may regulate the function of HPMCs through exosomal ITGA5B1/AEP, which may be crucial for peritoneal metastasis.

Role and mechanism of nursing cooperation and tetramethylpyrazine application in post-operative pain in patients undergoing total knee arthroplasty.

The aim of the present study was to investigate the effect of nursing cooperation on the post-operative complication of pain following total knee arthroplasty (TKA) and to explore the effects of tetramethylpyrazine (TMP) application on post-TKA pain. A total of 26 patients who received TKA between June 2014 and March 2016 were enrolled in this study. Nursing cooperation was provided to the patients during the TKA surgery, and pain was evaluated based on the visual analog scale (VAS). In addition, 40 male Sprague Dawley rats were used for the TKA model construction. The rats were randomly separated into 4 groups (sham, TKA, TKA+TMP and TKA+TMP+Interferon γ). Pain tolerance in rats was evaluated by mechanical stimulation. Inflammatory cytokine levels in TKA rat tissue were detected using ELISA. mRNA and protein expression of Janus kinase/signal transducer and activator of transcription 3 (JAK/STAT3) was detected using reverse transcription-polymerase chain reaction and western blot analysis, respectively. The results indicated that nursing cooperation serves a critical function during TKA and was associated with a lower level of pain compared with the control (P<0.05). Furthermore, TMP treatment reduced the level of inflammatory cytokines in the rat tissues, including interleukin (IL)-6, IL-10 and tumor necrosis factor-α in post-TKA (P<0.01). TMP was indicated to alleviate pain in post-TKA through suppressing the JAK/STAT3 signaling pathway. The results of the present study suggest that nursing cooperation is critical to TKA, and TMP may alleviate post-TKA pain through inhibiting the JAK/STAT3 signaling pathway.

[Effect of LINE1-ORF1p overexpression on the proliferation of nephroblastoma WT_CLS1 cells].

OBJECTIVE: To prepare the LINE1-ORF1p polyclonal antibody, and to study the effect of LINE1-ORF1p on the proliferation of nephroblastoma WT_CLS1 cells. METHODS: A genetic engineering method was used to achieve prokaryotic expression of LINE1-ORF1p, and rabbits were immunized with LINE1-ORF1p to prepare polyclonal antibody. Indirect ELISA was used to evaluate antibody titer, and Western blot and immunohistochemistry were used to evaluate the specific ability of antibody to recognize LINE1-ORF1p. The eukaryotic expression vector pEGFP-N1-LINE1-ORF1 was constructed and used to transfect WT_CLS1 cells. Western blot and qRT-PCR were used to measure the protein and mRNA expression of LINE1-ORF1, respectively, and cell proliferation assay and colony-forming assay were used to evaluate the effect of LINE1-ORF1p on the proliferation of WT_CLS1 cells and the formation of tumor cell clone. RESULTS: The LINE1-ORF1p antibody prepared had a titer of >1:16 000 and could specifically recognize LINE1-ORF1p in cells and tumor tissue. WT_CLS1 cells transfected with pEGFP-N1-LINE1-ORF1 had significant increases in the mRNA and protein expression of LINE1-ORF1 and significantly enhanced cell proliferation ability and colony formation ability (P<0.05). CONCLUSIONS: LINE1-ORF1p can promote the growth of nephroblastoma cells and the formation of tumor cell clone, and may be involved in the pathogenesis of nephroblastoma.