ABSTRACT
BACKGROUND: Reducing avoidable hospital admissions is a global healthcare priority, with optimal primary care recognised as pivotal for achieving this objective. However, in developing systems like China, where primary care is evolving without compulsory gatekeeping, the relationship between patient-perceived primary care quality and hospital utilisation remains underexplored. OBJECTIVES: This study aimed to explore the association between patient-perceived primary care quality and self-reported hospital utilisation in China. METHODS: Data were collected from 16 primary care settings. Patient-perceived quality of primary care was measured using the Assessment Survey of Primary Care scale across six domains (first-contact care, continuity, comprehensiveness, accessibility, coordination, and patient-centredness). Hospital utilisation included patient self-reported outpatient visits, hospital admissions, and emergency department (ED) visits in the last six months. Logistic regression analyses were examined associations between self-reported hospital utilisation and perceived primary care quality adjusted for potential confounders. RESULTS: Of 1,185 patients recruited, 398 (33.6%) reported hospital utilisation. Logistic regression analyses showed that higher total scores for patient-perceived quality of primary care were associated with decreased odds of hospital utilisation (adjusted odds ratio(AOR): 0.417, 95% confidence interval (CI): 0.308-0.565), outpatient visits (AOR: 0.394, 95% CI: 0.275-0.566) and hospital admissions (AOR: 0.496, 95% CI: 0.276-0.891). However, continuity of care was positively associated with ED visits (AOR: 2.252, 95% CI: 1.051-4.825). CONCLUSION: Enhanced patient-perceived quality of primary care in China is associated with a reduction in self-reported overall hospital utilisation, including outpatient visits and hospital admissions. However, better continuity of care may be associated with increased ED visits. Further research is warranted for precise insights and validation of these findings.
Improved patient-perceived primary care quality in China was linked to lower self-reported hospital utilisation, including outpatient visits and hospital admissions.Better primary care accessibility was associated with decreased self-reported emergency department visits and outpatient visits.Continuity of care showed a positive association with emergency department visits, warranting further research.
Subject(s)
Hospitals , Primary Health Care , Humans , Self Report , Cross-Sectional Studies , ChinaABSTRACT
Oral squamous cell carcinoma (OSCC) has become a global health problem due to its increasing incidence and high mortality rate. Early intervention through monitoring of the diagnostic biomarker levels during OSCC treatment is critical. Extracellular vesicles (EVs) are emerging surrogates in intercellular communication through transporting biomolecule cargo and have recently been identified as a potential source of biomarkers such as phosphoproteins for many diseases. Here, we developed a multiple reaction monitoring cubed (MRM3) method coupled with a novel sample preparation strategy, extracellular vesicles to phosphoproteins (EVTOP), to quantify phosphoproteins using a minimal amount of saliva (50 µL) samples from OSCC patients with high specificity and sensitivity. Our results established differential patterns in the phosphopeptide content of healthy, presurgery, and postsurgery OSCC patient groups. Notably, we discovered significantly increased salivary phosphorylated alpha-amylase (AMY) in the postsurgery group compared to the presurgery group. We hereby present the first targeted MS method with extremely high sensitivity for measuring endogenous phosphoproteins in human saliva EVs.
Subject(s)
Carcinoma, Squamous Cell , Extracellular Vesicles , Head and Neck Neoplasms , Mouth Neoplasms , Humans , Carcinoma, Squamous Cell/diagnosis , Biomarkers, Tumor/analysis , Saliva/chemistry , Mouth Neoplasms/diagnosis , Extracellular Vesicles/pathology , Squamous Cell Carcinoma of Head and Neck , Phosphoproteins/analysisABSTRACT
Extracellular vesicles (EVs) from biofluids have recently gained significant attention in the field of liquid biopsy. Released by almost every type of cell, they provide a real-time snapshot of host cells and contain a wealth of molecular information, including proteins, in particular those with post-translational modifications (PTMs) such as phosphorylation, as the main player of cellular functions and disease onset and progression. However, the isolation of EVs from biofluids remains challenging due to low yields and impurities from current EV isolation methods, making the downstream analysis of EV cargo, such as EV phosphoproteins, difficult. Here, we describe a rapid and effective EV isolation method based on functionalized magnetic beads for EV isolation from biofluids such as human urine and downstream proteomics and phosphoproteomics analysis following EV isolation. The protocol enabled a high recovery yield of urinary EVs and sensitive profiles of EV proteome and phosphoproteome. Furthermore, the versatility of this protocol and relevant technical considerations are also addressed here.
Subject(s)
Extracellular Vesicles , Proteomics , Humans , Proteomics/methods , Extracellular Vesicles/metabolism , Phosphoproteins/metabolism , Protein Processing, Post-Translational , Proteome/analysisABSTRACT
Extracellular vesicles (EVs) have emerged as a promising source of disease biomarkers for noninvasive early stage diagnoses, but a bottleneck in EV sample processing restricts their immense potential in clinical applications. Existing methods are limited by a low EV yield and integrity, slow processing speeds, low sample capacity, and poor recovery efficiency. We aimed to address these issues with a high-throughput automated workflow for EV isolation, EV lysis, protein extraction, and protein denaturation. The automation can process clinical urine samples in parallel, resulting in protein-covered beads ready for various analytical methods, including immunoassays, protein quantitation assays, and mass spectrometry. Compared to the standard manual lysis method for contamination levels, efficiency, and consistency of EV isolation, the automated protocol shows reproducible and robust proteomic quantitation with less than a 10% median coefficient of variation. When we applied the method to clinical samples, we identified a total 3,793 unique proteins and 40,380 unique peptides, with 992 significantly upregulated proteins in kidney cancer patients versus healthy controls. These upregulated proteins were found to be involved in several important kidney cancer metabolic pathways also identified with a manual control. This hands-free workflow represents a practical EV extraction and profiling approach that can benefit both clinical and research applications, streamlining biomarker discovery, tumor monitoring, and early cancer diagnoses.
Subject(s)
Extracellular Vesicles , Kidney Neoplasms , Humans , Workflow , Proteomics/methods , Proteins/analysis , Extracellular Vesicles/chemistry , Extracellular Vesicles/metabolismABSTRACT
Reversible protein S-nitrosylation regulates a wide range of biological functions and physiological activities in plants. However, it is challenging to quantitively determine the S-nitrosylation targets and dynamics in vivo. In this study, we develop a highly sensitive and efficient fluorous affinity tag-switch (FAT-switch) chemical proteomics approach for S-nitrosylation peptide enrichment and detection. We quantitatively compare the global S-nitrosylation profiles in wild-type Arabidopsis and gsnor1/hot5/par2 mutant using this approach, and identify 2,121 S-nitrosylation peptides in 1,595 protein groups, including many previously unrevealed S-nitrosylated proteins. These are 408 S-nitrosylated sites in 360 protein groups showing an accumulation in hot5-4 mutant when compared to wild type. Biochemical and genetic validation reveal that S-nitrosylation at Cys337 in ER OXIDOREDUCTASE 1 (ERO1) causes the rearrangement of disulfide, resulting in enhanced ERO1 activity. This study offers a powerful and applicable tool for S-nitrosylation research, which provides valuable resources for studies on S-nitrosylation-regulated ER functions in plants.
Subject(s)
Arabidopsis Proteins , Arabidopsis , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Nitric Oxide/metabolism , Arabidopsis/genetics , Arabidopsis/metabolism , Cysteine/metabolismABSTRACT
Many biological processes are regulated through dynamic protein phosphorylation. Monitoring disease-relevant phosphorylation events in circulating biofluids is highly appealing but also technically challenging. We introduce here a functionally tunable material and a strategy, extracellular vesicles to phosphoproteins (EVTOP), which achieves one-pot extracellular vesicles (EVs) isolation, extraction, and digestion of EV proteins, and enrichment of phosphopeptides, with only a trace amount of starting biofluids. EVs are efficiently isolated by magnetic beads functionalized with TiIV ions and a membrane-penetrating peptide, octa-arginine R8 + , which also provides the hydrophilic surface to retain EV proteins during lysis. Subsequent on-bead digestion concurrently converts EVTOP to TiIV ion-only surface for efficient enrichment of phosphopeptides for phosphoproteomic analyses. The streamlined, ultra-sensitive platform enabled us to quantify 500â unique EV phosphopeptides with only a few µL of plasma and over 1200â phosphopeptides with 100â µL of cerebrospinal fluid (CSF). We explored its clinical application of monitoring the outcome of chemotherapy of primary central nervous system lymphoma (PCNSL) patients with a small volume of CSF, presenting a powerful tool for broad clinical applications.
Subject(s)
Extracellular Vesicles , Phosphopeptides , Humans , Phosphopeptides/metabolism , Extracellular Vesicles/chemistry , Proteome/metabolism , Phosphoproteins/metabolismABSTRACT
Translating the research capability and knowledge in cancer signaling into clinical settings has been slow and ineffective. Recently, extracellular vesicles (EVs) have emerged as a promising source for developing disease phosphoprotein markers to monitor disease status. This study focuses on the development of a robust data-independent acquisition (DIA) using mass spectrometry to profile urinary EV phosphoproteomics for renal cell cancer (RCC) grades differentiation. We examined gas-phase fractionated library, direct DIA (library-free), forbidden zones, and several different windowing schemes. After the development of a DIA mass spectrometry method for EV phosphoproteomics, we applied the strategy to identify and quantify urinary EV phosphoproteomes from 57 individuals representing low-grade clear cell RCC, high-grade clear cell RCC, chronic kidney disease, and healthy control individuals. Urinary EVs were efficiently isolated by functional magnetic beads, and EV phosphopeptides were subsequently enriched by PolyMAC. We quantified 2584 unique phosphosites and observed that multiple prominent cancer-related pathways, such as ErbB signaling, renal cell carcinoma, and regulation of actin cytoskeleton, were only upregulated in high-grade clear cell RCC. These results show that EV phosphoproteome analysis utilizing our optimized procedure of EV isolation, phosphopeptide enrichment, and DIA method provides a powerful tool for future clinical applications.
Subject(s)
Carcinoma, Renal Cell , Extracellular Vesicles , Kidney Neoplasms , Humans , Carcinoma, Renal Cell/metabolism , Chromatography, Affinity/methods , Signal Transduction , Kidney Neoplasms/metabolism , Extracellular Vesicles/metabolismABSTRACT
Background: Measuring quality of primary care has attracted much attention around the world. Our team has developed and validated an Assessment Survey of Primary Care (ASPC) for evaluating quality of primary care in China. To facilitate the daily use of ASPC, this study aimed to develop and validate a rapid assessment version of ASPC (RA-ASPC) in China. Methods: This is a multi-phase study on 21 experts and 1,184 patients from 12 primary care facilities in 10 cities in China. Importance, representativeness, easy understanding, and general applicability of each item in ASPC scale were rated to select the top two ranked items for constituting RA-ASPC. Reliability of RA-ASPC was tested by calculating both Cronbach's alpha and McDonald's omega coefficients. Structural validity was assessed by exploratory and confirmatory factor analysis (EFA and CFA). Concurrent validity was performed by analyzing the relationship between RA-ASPC and patient satisfaction. Discriminant validity was tested by assessing the difference of RA-ASPC scores between patients with or without family doctors. Results: Ten items were selected for RA-ASPC. Both Cronbach's alpha (0.732) and McDonald's omega (0.729) suggested satisfactory internal consistency. In EFA, explained variance of RA-ASPC (72.6%) indicated its ability to measure quality of primary care in China. CFA indicators showed convincing goodness-of-fit (GFI = 0.996, AGFI = 0.992, CFI = 1.000, NFI = 0.980, RMR = 0.022, and the RMSEA = 0.000) for RA-ASPC. Positive association between RA-ASPC and patient satisfaction supported the concurrent validity of RA-ASPC. Patients with family doctors perceived higher quality of primary care than those without family doctors, indicating good discriminant validity of RA-ASPC. Conclusion: The theoretical framework of RA-ASPC was in line with internationally recognized core functions of primary care. Good psychometric properties of RA-ASPC proved its appropriateness in assessing quality of primary care from patients' perspectives in China.
Subject(s)
Primary Health Care , China , Humans , Psychometrics , Reproducibility of Results , Surveys and QuestionnairesABSTRACT
BACKGROUND: Patient-centred care is a core attribute of primary care. Not much is known about the relationship between patient-centred care and doctor-patient familiarity. This study aimed to explore the association between general practitioner (GP) perceived doctor-patient familiarity and the provision of patient-centred care during GP consultations. METHODS: This is a direct observational study conducted in eight community health centres in China. Level of familiarity was rated by GPs using a dichotomized variable (Yes/No). The provision of patient-centred care during GP consultations was measured by coding audiotapes using a modified Davis Observation Code (DOC) interactional instrument. Eight individual codes in the modified DOC were selected for measuring the provision of patient-centred care, including 'family information', 'treatment effects', 'nutrition guidance', 'exercise guidance', 'health knowledge', 'patient question', 'chatting', and 'counseling'. Multivariate analyses of covariance were adopted to evaluate the association between GP perceived doctor-patient familiarity and patient-centred care. RESULTS: A total of 445 audiotaped consultations were collected, with 243 in the familiar group and 202 in the unfamiliar group. No significant difference was detected in overall patient-centred care between the two groups. For components of patient-centred care, the number of intervals (1.36 vs 0.88, p = 0.026) and time length (7.26 vs. 4.40 s, p = 0.030) that GPs spent in 'health knowledge', as well as time length (13.0 vs. 8.34 s, p = 0.019) spent in 'patient question' were significantly higher in unfamiliar group. The percentage of 'chatting' (11.9% vs. 7.34%, p = 0.012) was significantly higher in the familiar group. CONCLUSIONS: This study suggested that GP perceived doctor-patient familiarity may not be associated with GPs' provision of patient-centred care during consultations in the context of China. Not unexpectedly, patients would show more health knowledge and ask more questions when GPs were not familiar with them. Further research is needed to confirm and expand on these findings.
Subject(s)
General Practitioners , China , Humans , Physician-Patient Relations , Primary Health Care , Referral and ConsultationABSTRACT
BACKGROUND: General practitioner (GP) consultation has long been considered an important component of general practice, but few studies have focused on its characteristics in China. OBJECTIVE: This study aimed to explore the content and elucidate the characteristics of GP consultations in general practice in China. METHODS: A multimethod investigation of GP consultations in eight community health centres in Guangzhou and Shenzhen, China was conducted between July 2018 and January 2019. Data from 445 GP consultations were collected by direct observation and audio tape and analysed by a modified Davis Observation Code with indicators for frequencies and detailed time durations. GP and patient characteristics were collected by post-visit surveys. RESULTS: The mean visit duration was approximately 5.4 minutes. GPs spent the most time on treatment planning, history taking, negotiating, notetaking and physical examination and less time on health promotion, family information collecting, discussing substance use, procedures and counselling. The time spent on procedures ranked first (66 seconds), followed by history taking (65 seconds) and treatment planning (63 seconds). Besides, patients were very active in the consultation, specifically for topics related to medicine ordering and drug costs. CONCLUSIONS: This study described the profile of GP consultations and illustrated the complexity of care provided by GPs in China. As patient activation in GP consultations becomes increasingly important, future studies need to explore how to promote the engagement of patients in the whole consultation process other than just requesting for medicine.
Subject(s)
General Practitioners , China , Family Practice , Humans , Physician-Patient Relations , Referral and ConsultationABSTRACT
BACKGROUND: Since 2009, the Chinese government has been reforming the healthcare system and has committed to reinforcing increased use of primary care. To date, however, the Chinese healthcare system is still heavily reliant on hospital-based specialty care. Studies consistently show an association between primary care and improved health outcomes, and the same association is also found among the disadvantaged population. Due to the "hukou" system, interprovincial migrants in the urban districts are put in a weak position and become the disadvantaged. Therefore, the aim of this study is to investigate whether greater supply and utilization of primary care was associated with reduced child mortality among the entire population and the interprovincial migrants in urban districts of Guangdong province, China. METHODS: An ecological study was conducted using a 3-year panel data with repeated measurements within urban districts in Guangdong province from 2014 to 2016, with 178 observations in total. Multilevel linear mixed effects models were applied to explore the associations. RESULTS: Higher visit proportion to primary care was associated with reductions in the infant mortality rate and the under-five mortality rate in both the entire population and the interprovincial migrants (p < 0.05) in the full models. The association between visit proportion to primary care and reduced neonatal mortality rate was significant among the entire population (p < 0.05) while it was insignificant among the interprovincial migrants (p > 0.05). CONCLUSIONS: Our ecological study based in urban districts of Guangdong province found consistent associations between higher visit proportion to primary care and improvements in child health among the entire population and the interprovincial migrants, suggesting that China should continue to strengthen and develop the primary care system. The findings from China adds to the previously reported evidence on the association between primary care and improved health, especially that of the disadvantaged.
Subject(s)
Child Health/statistics & numerical data , Community Health Services/supply & distribution , Community Health Services/statistics & numerical data , Patient Acceptance of Health Care/statistics & numerical data , Primary Health Care/statistics & numerical data , Child Mortality/trends , Child, Preschool , China/epidemiology , Datasets as Topic , Delivery of Health Care/organization & administration , Hospitals , Humans , Infant , Infant Mortality/trends , Transients and Migrants/statistics & numerical data , Urban Population/statistics & numerical dataABSTRACT
BACKGROUND: The Chinese government is ambitious regarding strengthening the primary care system for women and children. Primary care contributes to better health outcomes among neonates, infants, children and pregnant women, especially for vulnerable groups. However, few published studies have examined this issue in China. OBJECTIVE: This study examined whether greater supply and utilization of primary care was associated with improved health outcomes among targeted populations in the total and interprovincial migrant populations in the rural counties of Guangdong Province, China. METHODS: This ecological study analysed annual panel data from all 63 rural counties in Guangdong Province from 2014 to 2016 (n = 189). A linear random-effects panel data model was applied. RESULTS: Higher proportions of primary care visits were significantly associated with reduced incidences of low birth weight (P < 0.05) and preterm birth rates (P < 0.05) for the total population, and were significantly associated with reduced infant (P < 0.1) and under-five (P < 0.01) mortality rates for migrants. Greater primary care physician supply was significantly associated with reduced maternal mortality (P < 0.1) rates among migrants. However, primary care indicators were insignificant for both the total and migrant populations regarding neonatal mortality rates, as well as the infant and under-five mortality rates in the total population (P > 0.1). CONCLUSIONS: These findings support existing evidence regarding associations between primary care and improved health outcomes among newborns, children and pregnant women, especially for disadvantaged populations. However, associations were not significant for all studied health outcomes, implying the need for further study.
Subject(s)
Maternal Health , Premature Birth , Child , China/epidemiology , Female , Humans , Infant , Infant, Newborn , Pregnancy , Primary Health Care , Rural PopulationABSTRACT
In recent years, the incidence and mortality of cancer have witnessed a dramatic increase. Cancer has already caused severe economic burdens on society, especially in developing countries and has become a major public health concern. This study evaluates the medical economic burden, including total current curative expenditure (CCE) and catastrophic health expenditure (CHE) on cancer in Liaoning Province, China. A total of 252 medical institutions were investigated with multistage stratified cluster random sampling. We established a standardized database of 3 532 517 samples. "System of Health Account 2011", a new internationally recognized accounting system, was established to analyze the CCE on six most common cancers. CHE were estimated from the extracted 1344 patients with cancer, which performed a cross-sectional study. The association of individual and contextual factors with CHE was evaluated using logistic regression models. CCE for all the patients with the six types of cancer was 2801.38 million CNY in Liaoning Province, the highest of which was lung cancer. The incidence of CHE was 42.78%, while the threshold was 40%. The average and relative distance were 10.41% and 24.32%, respectively. Influencing factors were length of stay, type of health insurance, location of household, etc. Our findings highlight the need to address medical economic burden in the cancer population. Households with the cancer are more likely to incur CHE. Financial intervention to prevent it should target on poor households. We provide suggestions in aspects of health insurance and health service management to reduce CHE.