Influence of plasma collection tubes on N-glycome in human blood samples.

Background and aims: Quantitative analysis of plasma N-glycome is a promising method for identifying disease biomarkers. This study aimed to investigate the impact of using blood collection tubes with different anticoagulants on plasma N-glycome. Materials and methods: We used a robust mass spectrometry method to profile plasma N-glycomes in two cohorts of healthy volunteers (cohort 1, n = 16; cohort 2, n = 53). The influence of three commonly used blood collection tubes on fully characterized N-glycomic profiles were explored. Results: Principal component analysis revealed distinct clustering of blood samples based on the collection tubes. Pairwise comparisons demonstrated significant differences between EDTA and heparin plasma in 55 out of 82 quantified N-glycan traits, and between EDTA and citrate plasma in 62 out of 82 traits. These differences encompassed various N-glycan features, including glycan type, sialylation, galactosylation, fucosylation, and bisection. Trends in N-glycan variations in citrate and heparin plasma were largely consistent compared to EDTA plasma. In correlation analysis (EDTA vs. heparin; EDTA vs. citrate), Pearson's correlation coefficients were consistently higher than 0.7 for the majority of N-glycan traits. Conclusion: Sample matrix variations impact plasma N-glycome measurements. Caution is crucial when comparing samples from different plasma collection tubes in glycomics projects.

Bacterial outer membrane vesicles in cancer: Biogenesis, pathogenesis, and clinical application.

Outer membrane vesicles (OMVs) are spherical, nano-sized particles of bilayer lipid structure secreted by Gram-negative bacteria. They contain a series of cargos from bacteria and are important messengers for communication between bacteria and their environment. OMVs play multiple roles in bacterial survival and adaptation and can affect host physiological functions and disease development by acting on host cell membranes and altering host cell signaling pathways. This paper summarizes the mechanisms of OMV genesis and the multiple roles of OMVs in the tumor microenvironment. Also, this paper discusses the prospects of OMVs for a wide range of applications in drug delivery, tumor diagnosis, and therapy.

Phenotypic and genetic characteristics of 130 patients with mucopolysaccharidosis type II: A single-center retrospective study in China.

Background: Mucopolysaccharidosis Type II (MPS II) is a rare, progressive and ultimately fatal X-linked lysosomal storage disorder caused by mutations in the iduronate-2-sulfatase (IDS) gene. This report conducted a retrospective analysis to investigate the clinical characteristics, genotypes and management strategies in a large cohort of Chinese patients with MPS II. Methods: In this study, we explored 130 Chinese patients with MPS II between September 2008 and April 2022. Clinical manifestations, auxiliary examination, IDS pathogenic gene variants and IDS enzyme activity, surgical history were analysed in the study. Results: A total of 130 patients were enrolled and the mean age at diagnosis was 5 years old. This study found the most common symptoms in our patients were claw-like hands, followed by coarse facial features, birthmarks (Mongolian spot), delayed development, inguinal or umbilical hernia. The most commonly cardiac manifestations were valve abnormalities, which were mitral/tricuspid valve regurgitation (71.9%) and aortic/pulmonary valve regurgitation (36.8%). We had found 43 different IDS pathogenic gene variants in 55 patients, included 16 novel variants. The variants were concentrated in exon 9 (20% = 11/55), exon 3 (20% = 11/55) and exon 8 (15% = 8/55). A total of 50 patients (38.5%) underwent surgical treatment, receiving a total of 63 surgeries. The average age of first surgery was 2.6 years, and the majority of surgery (85.7%, 54/63) was operated before 4 years old. The most common and earliest surgery was hernia repair. Three patients were died of respiratory failure. Conclusion: This study provided additional information on the clinical, cardiac ultrasound and surgical procedure in MPS II patients. Our study expanded the genotype spectrum of MPS II. Based on these data, characterization of MPS II patients group could be used to early diagnosis and treatment of the disease.

Clinical nurses' moral courage and related factors: an empowerment perspective.

BACKGROUND: Moral courage as part of the moral competence of nurses has received increasing attention. Determination of the factors affecting moral courage is important in improving the quality of care. The purpose of this study was to investigate moral courage and related factors among frontline nurses from an empowerment perspective. METHODS: A cross-sectional study was conducted using data collection instruments comprising four main parts: the self-designed form of demographic characteristics, Conditions for Work Effectiveness II (CWEQ-II), Spreitzer's Psychological Empowerment Scale (PES) and Nurses' Moral Courage Scale (NMCS). Data were collected from 226 nurses in a tertiary hospital between February and March 2022 in Wuhan, the capital city of Hubei Province in central China. Descriptive statistics and multiple linear regression were used to analyze the data. RESULTS: The means of the total scores for the CWEQ-II, PES and the NMCS were 3.52 (SD = 0.69), 3.85 (SD = 0.53) and 3.90 (SD = 0.67), respectively. All the dimensions and the total scores of the CWEQ-II and PES were significantly correlated with the NMCS (p < 0.001). According to the multivariate stepwise regression analysis, CWEQ-II and PES were determined to be factors affecting NMCS. These variables explained 35.9% of the total variance in the moral courage scores of nurses. CONCLUSION: The level of moral courage among nurses is above average. Structural empowerment and psychological empowerment were the key factors affecting the promotion of moral courage. Hospital and organizational administrations should be conscious of the role of attach structural empowerment and psychological empowerment in the nursing workplace in increasing moral courage.

Application of patient decision aids in treatment selection of cardiac surgery patients: a scoping review.

BACKGROUND: The choice of treatment is an unavoidable challenge faced in the day to day medical decision making pertaining to patients with organic heart disease. As a professional discipline, cardiac surgery focuses on creating and using the most advanced evidence-based patient decision aids (PtDAs) to achieve high-quality decision-making. OBJECTIVES: To describe the basic situation, influencing factors, and the outcome of indicators of PtDAs among cardiac surgery patients. METHODS: Seven electronic databases were systematically searched for relevant reviews on the application of PtDAs among cardiac surgery patients. The methodological framework proposed by Arskey and O'Malley was used to guide the scoping review. The extracted data was analyzed qualitatively and quantitatively. RESULTS: After dual, blinded screening of titles and abstracts, 12 articles were included in the review. 10 were quantitative studies, 1 was a mixed study, 1 was a qualitative study. CONCLUSIONS: Compared with the burden of heart disease and the huge evidence base, the application of PtDAs in cardiac surgery is obviously insufficient. The published literature mainly provide information about the factors to be solved from the perspective of researchers, and also summarize obstacle factors. This is the basis for the application and construction of PtDAs in cardiac surgery patients.

Influence of Foot Ulceration on All-Cause and Cardiovascular Mortality in Diabetic Patients: A Case-Control Study.

PURPOSE: The purpose of this study was to analyze the influence of a diabetic foot ulcer on all-cause and cardiovascular disease (CVD) mortality. DESIGN: Retrospective case-control study. SUBJECTS AND SETTING OUTPATIENTS: Eighty-eight patients with new-onset diabetic foot ulceration (DFU) were paired with 176 patients without DFU (controls). The study setting was the Department of Endocrinology, Sun Yat-sen Memorial Hospital, located in Guangzhou, China. METHODS: Cause-specific mortality was recorded during a median follow-up duration of 6.20 years up to 1 March 2016. Records review dates were from January 1, 2004, to December 31, 2010. RESULTS: The all-cause mortality rate for the DFU group and the control group was 48.9% and 22.7%, respectively. The risk of all-cause death in the DFU group was 3.126 times higher than that in the control group (risk ratio [RR]= 3.126; 95% CI, 1.998-4.891; P = .000). The CVD mortality rate of the DFU group and the control group was 12.5% and 6.8%, respectively. The risk of CVD death in the DFU group was 3.277 times higher than that in the control group (RR = 3.277; 95% CI, 1.392-7.715; P = .007). CONCLUSIONS: Development of a diabetic foot ulcer was associated with a significantly higher all-cause and CVD-related death risk than that in a control group of persons with diabetes mellitus without DFU.

Comparative metagenomic analysis of the vaginal microbiome in healthy women.

The composition of these vaginal microbiome has a significant impact on women's health. However, few studies have characterized the vaginal microbiome of healthy Chinese women using metagenomic sequencing. Here, we carried out a comparative metagenomic analysis to survey taxonomic, functional levels, and microbial communities' genome content in healthy women's vaginal microbiome. Overall, we observed a total of 111 species, including all dominant vaginal Lactobacillus species, such as L. iners, L. crispatus, L. gasseri, and L. jensenii. Unlike microbial taxa, several pathways were ubiquitous and prevalent across individuals, including adenosine ribonucleotides de novo biosynthesis and pyruvate fermentation to acetate and lactate II. Notably, our diversity analysis confirmed a significant difference in healthy women from different ethnic groups. Moreover, we binned vaginal assemblies into 62 high-quality genomes, including 9 L. iners, 7 A. vaginae, 5 L. jensenii, and 5 L. crispatus. We identified the pan and core genomes of L. iners and A. vaginae and revealed the genetic diversity. Primary differences between strains were the hypothetical genes and mobile element-like genes. Our results provide a framework for understanding the implications of the female reproductive tract's composition and functional potential and highlight the importance of genome-resolved metagenomic analysis to further understand the human vaginal microbiome.

Urine Proteomics Profiling and Functional Characterization of Knee Osteoarthritis Using iTRAQ Technology.

Osteoarthritis (OA) is a degenerative chronic disease affecting the whole joint structures. With the increment in life expectancy and aging population, OA has become one of the largest socioeconomic burdens, associated with pain and loss of joint function. However, early laboratory tests of OA are still lacking. Therefore, new diagnostic tests for this disease are urgently needed. In this study, to gain an insight into the pathogenesis and the potential biomarkers of OA, we implemented a comparative urine proteomics study on OA patients and health people using iTRAQ-based mass spectrometry technology. Western blotting was used to validate the relative changes in urine protein levels for four of the identified proteins. We constructed a comprehensive urine proteome profile of the OA patients and identified 102 proteins differently changed in abundance. Forty-six proteins were upregulated and 56 proteins were significantly downregulated in OA patients. Furthermore, the proteins, COL-4, MMP9, adiponectin, and BBOX1 were validated through Western blots, which can serve as valuable candidate biomarkers and help to illustrate the pathogenesis of OA. These findings may provide clues for promising biomarkers for the early diagnosis and also offer a theoretical basis for the early treatment of OA.

Cobalt-Catalyzed Cross-Dehydrogenative C(sp2 )-C(sp3 ) Coupling of Oxazole/Thiazole with Ether or Cycloalkane.

Direct C5-alkylation of oxazole/thiazole with ether or cycloalkane has been achieved through a cobalt-catalyzed cross-dehydrogenative coupling (CDC) process in moderate to good yields. This transformation represents the first C(sp2 )-C(sp3 ) cross-coupling at the C5-position of the oxazole/thiazole via double C-H bond cleavages. Various functional groups on oxazole/thiazole substrates, as well as water and air, are well-tolerated with this concise and practical protocol, constituting straightforward access to heterocycles with great medicinal significance. A preliminary mechanism involving a radical process has also been proposed.

DDQ-promoted direct C5-alkylation of oxazoles with alkylboronic acids via palladium-catalysed C-H bond activation.

A novel and concise C5-alkylation of oxazoles using alkylboronic acids as alkyl donors via Pd(ii)-catalysed C-H bond activation has been achieved in moderate to good yields with satisfactory functional group tolerance. Instead of commonly used BQ as a key promoter, DDQ was discovered to be a better additive that significantly promoted this alkylation. This efficient and advanced method represents the first C(sp2)-C(sp3) cross-coupling reaction at the C5-position of oxazoles, which is particularly attractive due to its potential applications in the late-stage functionalization of oxazole-containing bioactive molecules.

Capsaicin mediates cell cycle arrest and apoptosis in human colon cancer cells via stabilizing and activating p53.

Capsaicin is the major pungent ingredient in red peppers which is world widely consumed. Except its potent pain relieving efficacy as reported, capsaicin also exerted its antitumor activity in several tumor models. Here, we reported that capsaicin had a profound anti-proliferative effect on human colon cancer cells via inducing cell cycle G0/G1 phase arrest and apoptosis, which was associated with an increase of p21, Bax and cleaved PARP. The underlying mechanism of capsaicin's antitumor potency was mainly attributed to the stabilization and activation of p53. Capsaicin substantially prolonged the half-life of p53 and significantly elevated the transcriptional activity of p53. Through suppressing the interaction between p53 and MDM2, MDM2-mediated p53 ubiquitination was remarkably decreased after capsaicin treatment, which resulted in the stabilization and accumulation of p53. The results of p53-shRNA experiment further demonstrated that p53 knockdown severely impaired the sensitivity of tested cells to capsaicin, G0/G1 phase arrest and the apoptosis induced by capsaicin in p53-knockdown cells was also dramatically decreased, implicating the important role of p53 played in capsaicin's antitumor activity. In summary, our data suggested that capsaicin, or a related analogue, may have a role in the management of human colon cancer.

Fast fixing and comprehensive identification to help improve real-time ligands discovery based on formaldehyde crosslinking, immunoprecipitation and SDS-PAGE separation.

BACKGROUND: Fast Fixation is necessary to study real-time protein-protein interactions under physiological conditions. Fast formaldehyde cross-linking can fix transient and weak protein interactions, thereby reducing the number of false negatives but producing great complexity. To reduce this complexity, immunoaffinity purification can Fish out complexes that include particular target proteins, but affinity-based co-purification has a limited capacity to eliminate nonspecific binding to beads and/or antibodies. To Filter out these complexes, SDS-PAGE is used to disrupt non-covalent bonds, thereby eliminating uncross-linked complexes and simultaneously providing molecular weight information for identification. RESULTS: We described a 4 F strategy to help improve real-time ligands discovery based on formaldehyde crosslinking, immunoprecipitation and SDS-PAGE separation: Fast Fix, Fish, and Filter, using albumin interactome as an example. The use of gel excision without staining makes this strategy comprehensive and sensitive. The target protein must be identified in the same slice as its ligands. The ligands must be identified in slices for the experimental group but not in the corresponding control slices. Only proteins that appear in the range of molecular weights equal to or greater than the sum of the proteins' theoretical molecular weights, together with the target, are considered ligands. In this study, 5 s of cross-linking with 10% formaldehyde was achieved in human blood. The use of this strategy identified 35 ligands for albumin. Comparison with four major previous studies of the albuminome revealed that 68.57% of the 35 ligands identified in our study were identified in these other studies. CONCLUSIONS: Fast cross-linking was achieved. The 4 F strategy can be used to identify real-time in situ interactions without prior intervention and to comprehensively identify ligands of particular target proteins with fewer false positives.

Identification and characterization of cow's milk proteins from the rat intestinal lymph using a proteomic strategy.

Food proteins were considered to be absorbed into the body after being digested to amino acids, dipeptides, and tripeptides. However, there are studies indicating that some proteins can pass through the intestinal epithelium under normal physiological conditions, perhaps not in sufficient quantities to be of nutritional importance, but in quantities that may be antigenically or biologically active. In the present study, rat intestinal lymph samples were collected using a modified lymph fistula rat model in fasting and cow's milk postprandial states. Low molecular weight proteins were enriched by ultrafiltration and differential solubilization, separated by 1D-SDS-PAGE, digested in-gel based on molecular weight, and identified using nano-LC-MS/MS. In the postprandial rat intestinal lymph, nine bovine-specific proteins (false discovery rate ≤1%) were identified in different molecular weight regions. Most proteins identified in lymph were highly abundant proteins in the milk, such as ß-lactoglobulin and caseins. Seven of the nine identified bovine-specific proteins are allergens in milk. This strategy can be used to search for proteins that can enter the intestinal lymph and analyze their common features. Understanding the common features of these proteins might help to develop protein drugs taken orally, so that therapeutic proteins might embody fusion domains for cross-barrier transport or translocation.

Differential protein expression in perfusates from metastasized rat livers.

BACKGROUND: Liver perfusates exhibit theoretical advantages regarding the discovery of disease biomarkers because they contain proteins that readily enter the blood-stream, and perfusion preserves the disease state in its natural context. The purpose of the study is to explore the value of liver perfusate proteome in the biomarker discovery of liver diseases. RESULTS: In this study, 86 differentially expressed proteins were identified in perfusates from isolated rat livers metastasized by Walker-256 tumor cells. Among these proteins, 27 were predicted to be secreted, and 59 were intracellular or membrane proteins. Most of the secretory proteins (70.4%) were decreased in metastasized liver perfusates. The main canonical ingenuity pathway to which these secretory proteins belonged was acute phase response, which indicated that the liver-associated immune reaction was damaged by the metastasis. In contrast, most of the intracellular or membrane proteins (86.4%) exhibited higher relative abundances in the metastasized liver perfusates. Some of these proteins, including Rpl21, Atic, Eif3s2, Echs1, Eps15 and Ywhab, have previously been reported to be involved in cancer genesis and progression. As a member of the 14-3-3 protein family, Ywhab plays a key role in cellular proliferation and oncogenic transformation and has been reported to be involved in the development of breast cancer. Its abundance was elevated by 3.5-fold in the metastasized perfusates. Validation by Western blotting revealed a 3.7-fold increase in the abundance of this protein in metastasized plasma. CONCLUSIONS: These results show that perfusate proteome can be used as an alternative initial resource for biomarker identification, which ultimately requires validation in serum.

A comparison of E15.5 fetus and newborn rat serum proteomes.

BACKGROUND: Serum proteins carry out several functions in the circulation, including transfer, immunological functions, messenger functions, coagulation, and regulation of homeostasis. To investigate changes in serum proteins that occur during development, the serum proteomes of embryonic 15.5 (E15.5) fetuses and newborn rats were compared using LC-MS/MS. RESULTS: A total of 958 proteins were identified in the serum of rats at both developmental stages. The serum proteome pattern of newborn rats was compared to E15.5 fetuses by relative quantitation. The expression patterns of hemoglobin subunits were different at the two stages, with most of the subunits having decreased expression in newborn rats compared to E15.5 fetuses. In addition, 8 of 12 apolipoproteins were significantly decreased and 10 of 11 identified complement molecules were increased, with 4 exhibiting a significant increase. Moreover, 11 of 14 of the significantly increased enzyme regulators were inhibitors. The serum proteome patterns of different littermates from both developmental stages were also compared. We found that the levels of many highly abundant serum proteins varied between littermates, and the variations were larger than the variations of the technical control. CONCLUSIONS: The serum proteomes of newborn rats and E15.5 fetuses were compared. The expression patterns of hemoglobin subunits were different at the two developmental stages, with most of the subunits having decreased expression. The majority of apolipoproteins had significantly decreased expression, while almost all identified complement proteins had increased expression. The levels of several highly abundant serum proteins also varied among littermates at these two developmental stages. This is the first study using LC-MS/MS to investigate serum proteome development.

A tool for biomarker discovery in the urinary proteome: a manually curated human and animal urine protein biomarker database.

Urine is an important source of biomarkers. A single proteomics assay can identify hundreds of differentially expressed proteins between disease and control samples; however, the ability to select biomarker candidates with the most promise for further validation study remains difficult. A bioinformatics tool that allows accurate and convenient comparison of all of the existing related studies can markedly aid the development of this area. In this study, we constructed the Urinary Protein Biomarker (UPB) database to collect existing studies of urinary protein biomarkers from published literature. To ensure the quality of data collection, all literature was manually curated. The website (http://122.70.220.102/biomarker) allows users to browse the database by disease categories and search by protein IDs in bulk. Researchers can easily determine whether a biomarker candidate has already been identified by another group for the same disease or for other diseases, which allows for the confidence and disease specificity of their biomarker candidate to be evaluated. Additionally, the pathophysiological processes of the diseases can be studied using our database with the hypothesis that diseases that share biomarkers may have the same pathophysiological processes. Because of the natural relationship between urinary proteins and the urinary system, this database may be especially suitable for studying the pathogenesis of urological diseases. Currently, the database contains 553 and 275 records compiled from 174 and 31 publications of human and animal studies, respectively. We found that biomarkers identified by different proteomic methods had a poor overlap with each other. The differences between sample preparation and separation methods, mass spectrometers, and data analysis algorithms may be influencing factors. Biomarkers identified from animal models also overlapped poorly with those from human samples, but the overlap rate was not lower than that of human proteomics studies. Therefore, it is not clear how well the animal models mimic human diseases.