Unified calibration of D-dimer can improve the uniformity of different detection systems.

Background: D-dimer at a low level is important evidence for excluding the onset and progression of thrombosis. It is readily detectable and yields rapid results, although significant variability exists among different detection systems. Our study aims to enhance the consistency across various detection systems. Methods: Twelve detection systems were included in our study. We sought to address this inconsistency by using various calibrators (two supplied by manufacturers and two comprising pooled human plasma diluted with different diluents) to standardize D-dimer measurements. We categorized the data into three groups according to D-dimer concentration levels: low (≤0.5 mg/L), medium (>0.5 mg/L - <3 mg/L), and high (≥3 mg/L). We then analyzed the data focusing on range, consistency, comparability, negative coincidence rate, and false negative rate. Results: Calibrating with pooled human plasma led to narrower result ranges in the low and medium groups (P < 0.05). In the low group, consistency improved from weak to strong (ICC 0.4-0.7, P﹤0.05), while it remained excellent in the other groups and overall (ICCï¹¥0.75, P﹤0.05). The percentage of pairwise comparability increased in both the low and high groups. Additionally, there was an increase in the negative coincidence rate. Conclusion: These findings demonstrate that uniform calibration of D-dimer can significantly enhance the consistency of results across different detection systems.

Protein profile of circulating extracellular vesicles reveals biomarker candidates for diagnosis of post-traumatic deep vein thrombosis.

BACKGROUND AND OBJECTIVE: Deep vein thrombosis (DVT) is a common complication after trauma and mostly without specific symptoms. Timely diagnosis and early appropriate treatment measures can prevent further development of thrombosis for patients with traumatic lower extremity fractures. Although extracellular vesicles (EVs) are confirmed as promising disease biomarkers, little is known about the role of altered levels and composition in the diagnosis of post-traumatic DVT. METHOD: The levels of circulating EVs subgroups were measured using flow cytometry. Isolated EVs were characterized and subjected to proteomics analysis to screen for differentially expressed proteins (DEPs) between DVT and non-DVT patients. Regularized logistic regression analysis based on L2 penalty terms using R's caret package was applied to build a model for DVT diagnosis. RESULTS: Compared to non-DVT patients, DVT patients had higher circulating hepatocyte-derived EVs (hEVs) with good predictive value for post-traumatic DVT diagnosis. The results of the proteomic analysis showed that differentially expressed proteins (DEPs) of circulating EVs between the DVT group and non-DVT group were enriched in the complement and coagulation cascade. Finally, an integrated model of five biomarkers including SERPING1, C8G, CFH, FIX, and hEVs level was established for post-traumatic DVT diagnosis with robust identification of the traumatic patients with and without DVT (AUC 0.972). CONCLUSION: Post-traumatic DVT patients had changed levels and composition of circulating EVs compared to non-DVT patients and healthy controls. Circulating EVs may acquire pathological protein signatures and become potential biomarkers for identifying subjects' post-traumatic DVT.

Relationship between patient-perceived quality of primary care and self-reported hospital utilisation in China: A cross-sectional study.

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.

Discovery of novel serum biomarkers for diagnosing and predicting postmenopausal osteoporosis patients by 4D-label free protein omics.

We aimed to identify protein biomarkers that could rapidly and accurately diagnose osteoporosis patients (OPs) using a highly sensitive proteomic immunoassay. Four-dimensional (4D) label-free proteomics analysis was performed to determine the differentially expressed proteins in serum collected from 10 postmenopausal osteoporosis patients and 6 non-osteoporosis patients. The ELISA method was used to select the predicted proteins for verification. Serum was taken from 36 postmenopausal osteoporosis patients and 36 healthy individuals from normal postmenopausal women. Receiver operating characteristic (ROC) curves were used to determine the diagnostic potential of this method. We validated the expression of these six proteins using ELISA. The CDH1, IGFBP2, and VWF of osteoporosis patients were significantly higher than those of the normal group. PNP was significantly lower than that in the normal group. And using ROC curve calculation, serum CDH1 had a cut-off of 3.78 ng/mL with a sensitivity of 84.4%, and PNP had a cut-off of 944.32 ng/mL with 88.9% sensitivity. These outcomes suggest that serum-level CHD1 and PNP have the potential power as effective indicators for the diagnosis of PMOP. Our results suggest that CHD1 and PNP might be associated with the pathogenesis of OP and would be helpful in diagnosing OP. Therefore, CHD1 and PNP may act as potential key markers in OP.

Determination of Vitamin K1, MK-4, MK-7, and D Levels in Human Serum of Postmenopausal Osteoporosis Women Based on High Stability LC-MS/MS: MK-7 May Be a New Marker of Bone Metabolism.

INTRODUCTION: Vitamin K (VK) as well as vitamin D (VD) plays an important role in osteoporosis. Vitamin K1 (VK1), vitamin K2 (VK2, menaquinone-4 (MK-4), and menaquinone-7 (MK-7)) are significant for the metabolism of skeletal muscle. 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 (25(OH)D2 and 25(OH)D3) reflect circulating VD levels. More sensitive measurements remain to be developed. In the present study, a new high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of VK1, VK2 (MK-4 and MK-7), as well as 25(OH)D2 and 25(OH)D3 levels in human serum. METHODS: We developed a simple LC-MS/MS method for the determination of VK1, MK-4, MK-7, 25(OH)D2, and 25(OH)D3 levels in human serum and validated the method in a study cohort of 200 patients divided into the premenopausal women group and postmenopausal osteoporosis patient group. RESULTS: The overall precision (coefficient of variation) ranged from 2.66 to 10.11% in the specified working range (0.05-5 ng/mL) for VK1, MK-4, and MK-7. Serum VK1, MK-4, and MK-7 levels in postmenopausal women with osteoporosis were 1.187 ± 0.094 ng/mL, 0.058 ± 0.009 ng/mL, and 0.885 ± 0.064 ng/mL, respectively (mean ± standard deviation). Serum VK1, MK-4, and MK-7 levels in premenopausal women were 1.143 ± 0.103 ng/mL, 0.028 ± 0.003 ng/mL, and 1.553 ± 0.226 ng/mL, respectively. Serum 25(OH)D2 and 25(OH)D3 levels in postmenopausal women with osteoporosis were 0.757 ± 0.056 ng/mL and 11.72 ± 0.632 ng/mL, respectively. Serum 25(OH)D2 and 25(OH)D3 levels in premenopausal were 1.793 ± 0.575 ng/mL and 12.42 ± 1.069 ng/mL, respectively. CONCLUSION: A new LC-MS/MS method for determination of serum VK and VD levels was evaluated and validated. MK-7 in plasma decreased earlier than VD in postmenopausal osteoporosis patients. MK-7 status is significantly associated with osteoporosis and could be considered a predictable biomarker in the diagnosis of osteoporosis in postmenopausal women.

Hepatocyte-derived Microparticles as Novel Biomarkers for the Diagnosis of Deep Venous Thrombosis in Trauma Patients.

Venous thromboembolism is a common complication following trauma. We investigated the dynamics of plasma microparticles (MPs) levels and explored their potential as biomarkers of deep vein thromboembolism (DVT) after trauma. A total of 775 patients with traumatic fractures were recruited in this nested study. About 106 trauma patients (53 DVT subjects and 53 age-, sex-, and fracture site-matched non-DVT subjects) and 53 healthy volunteers met the enrollment criteria. MPs were characterized by transmission electron microscope, nanoparticle tracking analysis, and western blotting. Circulating levels of MPs were measured using a flow cytometer. Meanwhile, routine laboratory parameters were examined in all patients. Compared to non-DVT patients, DVT patients had higher circulating phosphatidylserine (PS) + MPs, hepatocyte-derived MPs (HMPs), PS + HMPs, and platelet-derived MPs (PMPs). Notably, PS + HMPs had the best predictive value for DVT diagnosis in trauma patients (area under the curve [AUC] 0.8939, 95% CI 0.8326 to 0.9552), which was superior to d-dimer (AUC 0.5881). The Hepatic Procoagulant Index combined plasma levels of PS + HMPs and albumin, increasing the AUC to 0.8978 (95% CI 0.8396 to 0.9561). This is the first study that addressed circulating PS + HMPs are promising biomarkers with high performance in diagnosing DVT. The Hepatic Procoagulant Index is a potential predictor of DVT in trauma patients.

Development and Validation of a Rapid Assessment Version of the Assessment Survey of Primary Care in China.

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.

Association between doctor-patient familiarity and patient-centred care during general practitioner's consultations: a direct observational study in Chinese primary care practice.

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.

Involvement of TLR4 signaling regulated-COX2/PGE2 axis in liver fibrosis induced by Schistosoma japonicum infection.

BACKGROUND: Hepatic stellate cell (HSC) activation plays a pivotal role in hepatic inflammation and liver fibrosis. TLR4 pathway activation has been reported to be involved in mice liver fibrosis induced by hepatitis virus infection, alcohol abuse, biliary ligation, carbon tetrachloride 4 treatment, and Schistosoma japonicum (Sj) infection. The effect and mechanisms of the cyclooxygenase 2 (COX2)/prostanoid E2 (PGE2) axis on liver fibrosis induced by Sj are still unclear. METHODS: Mice liver fibrosis were induced by cutaneous infection of Sj cercariae. COX-2 inhibitor, NS398 were injected from week 5 to week 7, while TLR4 inhibitor TAK242 were injected from week 4 to week 8 post Sj infection. Human HSCs line, LX-2 cells were cultured and exposed to LPS or synthetic PGE2, or pretreated by TAK242, TLR4-siRNA or NS398. Liver tissue and serum or in vitro cultured cell lysaste were collected at indicated time courses for exploring the relationship between TLR4 and COX2-PGE2 axis through qPCR, western blot, immunohistochemical assay, ect. One-way analysis of variance among multiple groups followed by Uncorrected Fisher's LSD-t test or paired comparisons through t test were performed to tell the statistical differences. RESULTS: This study investigated the link between the COX2/PGE2 axis and TLR4 signaling in the induction of liver fibrogenesis in mice during Sj infection and in vitro culture of HSC strain-LX-2. The COX2/PGE2 axis was positively associated with Sj-induced liver fibrosis. TLR4 pathway activation stimulated the COX2/PGE2 axis in Sj-infected mice and in lipopolysaccharide (LPS)-exposed cultured HSCs. Synthetic PGE2 activated cultured HSCs through upregulation of alpha smooth muscle actin (α-SMA) expression. In LPS-triggered HSCs, NS398, a COX2 inhibitor, led to suppression of PGE2 synthesis and reduced expression of α-SMA and type I collagen (COL I). CONCLUSIONS: These results indicate firstly the positive association of the COX2/PGE2 axis with liver fibrosis induced by Sj infection. TLR4 signaling may at least partially control the COX2/PGE2 axis in Sj-infected mice liver and in vitro cultured HSCs. The COX2/PGE2-EP2/EP4 axis might be a good drug target against liver fibrosis induced by Sj infection.

Effect of Family Practice Contract Services on the Perceived Quality of Primary Care among Patients with Multimorbidity: A Cross-Sectional Study in Guangdong, China.

Family practice contract services, an important primary-care reform policy for improving primary healthcare quality in China, incorporate patients with multiple chronic conditions into the priority coverage groups and focus on their management. This study aims to explore the family practice contract services' effectiveness in improving the quality of primary care experienced by this population. A cross-sectional study using a three-stage sampling was conducted from January to March 2019 in Guangdong, China. A multivariable linear regression, including interaction terms, was applied to examine the associations between the contract services and primary care quality among people with different chronic conditions. The process quality of primary care was measured in six dimensions using the validated assessment survey of primary care (ASPC) scale. People with contract services scored higher in terms of quality of primary care than those without contract services. Contract services moderated the association between chronic condition status and primary care quality. Significantly positive interactions were observed in the patient-centred care dimension and negative interactions were reflected in the accessibility dimension. Our findings suggest that family practice contract services play a crucial role in improving patient-perceived primary care quality and provide emerging evidence that patients with multimorbidity tend to benefit more from the services, especially in patient-centred care.

Major Pollen Allergen Components and CCD Detection in Bermuda Grass Sensitized Patients in Guangzhou, China.

OBJECTIVE: Bermuda grass pollen is a common inhaled allergen. The aim of this study was to investigate the molecular sensitization patterns to major pollen allergens (Bermuda grass, Mugwort and Timothy grass) and cross-reactive carbohydrate determinants (CCD) in Bermuda grass sensitized patients in southern China. METHODS: Serum specific IgE (sIgE) levels of Bermuda grass allergen components (Cyn d 1 and Cyn d 12), Timothy grass allergen components (Phl p 1, Phl p 4, Phl p 5, Phl p 7 and Phl p 12), Mugwort allergen components (Art v 1, Art v 3 and Art v 4) and CCD were detected in 78 patients sensitized to Bermuda grass via EUROBlotMaster system. RESULTS: Compared with CCD-positive patients, those with negative CCD results had significant higher positive rates of Cyn d 1 (47.8% vs 14.5%), Phl p 1 (26.1% vs 7.3%), Phl p 12 (21.7% vs 3.6%) and Art v 4 (26.1% vs 3.6%) (all p < 0.05). Patients <18 years old had the highest positive rate of Cyn d 1 (40.7%). Additionally, rhinitis patients had the highest positive rate of Cyn d 1 (60.0%), and all patients with Cyn d 12 sensitization (17.2%) were asthmatic patients. Optimal scale analysis showed that Phl p 1 and Cyn d 1 were closely related (Cronbach's alpha = 85.1%). CONCLUSION: The highest positive rate of pollen allergen components was Cyn d 1 in Bermuda grass sensitized patients in southern China. Most patients were sensitized to CCD alone, and CCD may have less interference in the detection of Cyn d 1, Art v 4, Phl p 1 and Phl p 12. The sensitization patterns of pollen allergen components varied in different ages and diseases, and the diagnostic strategy of pollen allergen needs to be considered in the future.

An overview of general practitioner consultations in China: a direct observational study.

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.

The effects of gatekeeping on the quality of primary care in Guangdong Province, China: a cross-sectional study using primary care assessment tool-adult edition.

BACKGROUND: Developed countries have widely implemented a gatekeeping system as a core policy of primary care, also known as the system of first visit in the community. As gatekeepers, general practitioners are responsible for the diagnosis and treatment of residents in the community health centres, and referring patients to specialists as appropriate. After several years of healthcare reform, gatekeeping policy has achieved remarkable success in China. Shenzhen and Dongguan were the first batch of pilot cities that implemented the policy of gatekeeping. This study aims to examine the effects of gatekeeping on the quality of primary care between the gatekeeping and non-gatekeeping groups in these two pilot cities. METHODS: A cross-sectional survey was conducted in five community health centres in Shenzhen and Dongguan cities, both located within Guangdong Province, China, using a validated Chinese version of the Primary Care Assessment Tool-Adult Edition (PCAT-AE) and carrying out face-to-face interviews with patients 18 years and older. Analyses were grouped according to whether or not patients had gatekeepers. Propensity Score Matching was used to control for confounding factors. A chi-square test was used to compare the factors mentioned above and an independent t-test was performed to compare the eight domains of the core functions of primary care between the two groups of patients. RESULTS: In total, 765 valid questionnaires were collected for analysis, after matching the sample size were 238 pairs. All the confounding factors observed between the gatekeeping and non-gatekeeping groups were balanced. The PCAT-AE scores for first-contact utilisation (3.29 > 2.66, p < 0.001) and coordination (2.06 > 1.95, p < 0.05) were higher in the gatekeeping group after matching, but the domains of accessibility (1.59 < 1.67, p < 0.05) and continuity (2.26 < 2.40, p < 0.05) were lower. The PCAT-AE mean score was slightly higher in gatekeeping group (1.98 > 1.93, p > 0.05) but without statistical significance. CONCLUSION: This study demonstrated that gatekeeping has helped to improve first-contact utilisation and coordination of primary care, but that other goals such as continuity and comprehensiveness have been harmed. To establish a sustainable gatekeeping system and to strengthen the core functions of the community comprehensively, the current gatekeeping system needs refinement.

Positive Feedback Regulation between Transglutaminase 2 and Toll-Like Receptor 4 Signaling in Hepatic Stellate Cells Correlates with Liver Fibrosis Post Schistosoma japonicum Infection.

Liver fibrosis induced by Schistosoma japonicum (Sj) infection is characterized by the accumulation of extracellular matrix (ECM). The activated and differentiated hepatic stellate cells (HSCs) are the predominant ECM-producing cell type in the liver. Toll-like receptor (TLR) 4 pathway activation plays a key role in mice liver fibrosis models induced by alcohol, biliary ligation, and carbon tetrachloride 4. In this work, we found that TLR4 pathway activation correlated with the severity of liver fibrosis post Sj infection. The TLR4 receptor inhibitor TAK242 reduced the extent of liver fibrosis. The increased expression of TLR4, α-smooth muscle actin (α-SMA), and cytoglobin was observed in the HSCs of mouse liver after Sj infection. In response to stimulation with either lipopolysaccharide or Sj's soluble egg antigen (SEA), high levels of TLR4 and α-SMA were induced in HSCs and were inhibited by TAK242 treatment. In previous work, we had reported that a high level of transglutaminase 2 (TGM2) is crucial for liver fibrosis post Sj infection. Herein, we found that TLR4 signaling also controlled Tgm2 expression. Inhibition of TGM2 activity by cystamine (CTM) in Sj-infected mice or in HSCs induced with all-trans-retinoic acid (ATRA) stimulation led to a lowered activation of TLR4 signaling and a reduced α-SMA expression. These results were confirmed by downregulating the Tgm2 gene by specific siRNA. These observations implied the presence of a positive feedback regulation between TGM2 and TLR4 signaling in HSCs that correlated with liver fibrosis post Sj infection. This novel connection between TGM2 and TLR4 pathway activation in liver fibrosis induced by Sj infection enhances our understanding of liver diseases.