Optimization path of primary public health service talent team construction: a largescale survey in Huaihai Economic Zone, China.

Background: The primary public health service system is indispensable for the implementation of the "Healthy China 2030" strategy, and primary healthcare workers, as the key drivers of this system, play a pivotal role in its development and establishment to ensure population well-being. In developing countries, such as China, primary public health systems are still weak, and in order to address this phenomenon, health system reform is needed, and primary public health personnel are crucial to health system reform. The current situation of primary public health workers in low-income and developing countries is characterized by varying degrees of problems that need improvement. Objectives: The purpose of this study is to understand the current situation of primary public health service workforce building, analyze the existing problems of the workforce, put forward suggestions for improvement and explore countermeasures, and provide Chinese wisdom and a reference basis for primary public health workforce building in the world, especially in developing countries. Methods: Combining the Work-Family Conflict Scale, Copenhagen Burnout Inventory, Minnesota Satisfaction Questionnaire, and Turnover Intention Scale, a relevant survey questionnaire was designed to quantitatively investigate the baseline characteristics of primary public health service institutions and their staff in four representative cities in the Huaihai Economic Zone: Xuzhou in Jiangsu Province, Linyi in Shandong Province, Shangqiu in Henan Province, and Huaibei in Anhui Province. The collected data were analyzed and processed using SPSS 25.0 statistical analysis software through univariate analysis and logistic regression analyses. Methods such as one-way ANOVA, Logistic regression analysis, and independent samples t-test were used to analyze the influencing factors of primary public health workforce development. Results: The current work intensity at the primary public health level is currently high, the salary and benefits cannot meet the needs of most primary public health personnel, and the competition between work and family in terms of time and resources is pronounced, and the majority of primary public health personnel are dissatisfied with the status quo of "doing more work for less reward" and the poor social security. Emotional exhaustion, depersonalization, and a sense of personal accomplishment were positively correlated with the tendency to leave (all p < 0.01), and the burnout and emotional exhaustion of primary public health workers were intense. Conclusion: Primary public health personnel play an important role in providing primary public health services. However, the current working conditions of junior public health personnel in the Huaihai Economic Zone are influenced by factors such as workload, income level, and employment situation improvement, leading to low job satisfaction, significant work-family conflicts, and high turnover intention. In this context, based on the opinions of grassroots administrative departments and internationally relevant experiences, a series of suggestions have been proposed to improve the professional service level, job satisfaction, and occupational identity of staff members. These suggestions make valuable contributions to both the Huaihai Economic Zone and countries worldwide in safeguarding individual health and promoting national primary healthcare reform.

Effects of work-family conflict, social support and burnout on job satisfaction among primary care physicians in Huaihai economic zone.

Background: Primary care physicians (PCPs) are doctors in primary health care institutions, namely village clinics, township health centers and community health service centers (stations) who are the main providers of primary health care services in primary health care settings. Improving the overall health status of the population requires the support of a large number of primary care physicians; however, the job satisfaction of this group has not been sufficiently emphasized and recognized. Objective: The purpose of this study was to examine the effects of primary care physicians' work-family conflict on their job satisfaction, as well as the mediating role of burnout and the moderating role of social support. Methods: This cross-sectional study was conducted from February 2023 to March 2023. Participants were 749 primary care physicians from four cities of Xuzhou, Linyi, Huaibei, and Shangqiu in the Huaihai Economic Zone of China. SPSS statistical analysis was used to evaluate the relationship between work-family conflict, social support, burnout and job satisfaction among medical workers. Results: Work-family conflict had a significant negative effect on job satisfaction (ß = -0.36, p< 0.001), after adding burnout in the model, work-family conflict also negatively predicted job satisfaction (ß = -0.32, p< 0.001). Social support had a moderating effect on the direct effect of burnout on job satisfaction (ß = 0.00, t = 2.66, p< 0.01, 95%CI [0.001, 0.007]), the predictive effect of burnout on job satisfaction at high level of social support (ß = -0.45, p< 0.001) was higher than a low level of social support (ß = -0.33, p< 0.001). Conclusions: This study demonstrated the negative impact of work-family conflict on primary care physicians' job satisfaction, as well as the mediating role of burnout and the moderating role of social support on burnout and job satisfaction, which are important for improving primary care physicians' job satisfaction and enhancing the quality of primary care in the future.

Pyroptotic-Spatiotemporally Selective Delivery of siRNA against Pyroptosis and Autoimmune Diseases.

Small-interfering RNAs (siRNAs) offer promising prospects for treating pyroptosis-related autoimmune diseases. However, poor stability and off-target effects during in vivo transportation hinder their practical clinical applications. Precision delivery and adaptive release of siRNAs into inflamed tissues and immune cells could unleash their full therapeutic potential. This study establishes a pyroptotic-spatiotemporally selective siRNA delivery system (PMRC@siGSDME) that selectively targets inflammatory tissues, responds to pyroptosis, and exhibits remarkable therapeutic efficacy against various autoimmune diseases. Novel hybrid nanovesicles (NVs) are designed as a combination of pyroptotic macrophage membranes (PMs) and R8-cardiolipin-containing nanovesicles (RC-NVs). Evidence provides that PM-derived proteins involved in cell-cell interactions and membrane trafficking may contribute to the specificity of NVs to inflammatory tissue. In addition, cardiolipin anchored in the hybrid NVs increases its affinity for activated gasdermin E (GSDME) and achieves pyroptosis-adaptive release of siGSDME for the spatiotemporally selective suppression of immune responses. More importantly, PMRC@siGSDME displays significant anti-inflammatory and therapeutic effects in multiple mouse autoimmune disease models, including arthritis and inflammatory bowel disease (IBD). Collectively, an innovative siRNA delivery strategy precisely tailored for pyroptotic cells has been developed, paving the way for new treatments for autoimmune inflammatory diseases with minimal side effects and wide clinical applicability.

[Neutrophil extracellular traps extrusion from neutrophils stably adhered to ICAM-1 by lipoteichoic acid stimulation].

The effect of neutrophil extracellular traps (NETs) on promoting intravascular microthrombi formation and exacerbating the severity of sepsis in patients has gained extensive attention. However, in sepsis, the mechanisms and key signaling molecules mediating NET formation during direct interactions of endothelial cells and neutrophils still need further explored. Herein, we utilized lipoteichoic acid (LTA), a component shared by Gram-positive bacteria, to induce NET extrusion from neutrophils firmly adhered to the glass slides coated with intercellular adhesion molecule-1(ICAM-1). We also used Sytox green to label NET-DNA and Flou-4 AM as the intracellular Ca 2+ signaling indicator to observe the NET formation and fluctuation of Ca 2+ signaling. Our results illustrated that LTA was able to induce NET release from neutrophils firmly attached to ICAM-1-coated glass slides, and the process was time-dependent. In addition, our study indicated that LTA-induced NET release by neutrophils stably adhered to ICAM-1 depended on Ca 2+ signaling but not intracellular reactive oxygen species (ROS). This study reveals NET formation mediated by direct interactions between endothelial ICAM-1 and neutrophils under LTA stimulation and key signaling molecules involved, providing the theoretical basis for medicine development and clinical treatment for related diseases.

Structural and functional regulation of Chlamydomonas lysosome-related organelles during environmental changes.

Lysosome-related organelles (LROs) are a class of heterogeneous organelles conserved in eukaryotes that primarily play a role in storage and secretion. An important function of LROs is to mediate metal homeostasis. Chlamydomonas reinhardtii is a model organism for studying metal ion metabolism; however, structural and functional analyses of LROs in C. reinhardtii are insufficient. Here, we optimized a method for purifying these organelles from 2 populations of cells: stationary phase or overloaded with iron. The morphology, elemental content, and lysosomal activities differed between the 2 preparations, even though both have phosphorus and metal ion storage functions. LROs in stationary phase cells had multiple non-membrane-bound polyphosphate granules to store phosphorus. Those in iron-overloaded cells were similar to acidocalcisomes (ACs), which have a boundary membrane and contain 1 or 2 large polyphosphate granules to store more phosphorus. We established a method for quantifying the capacity of LROs to sequester individual trace metals. Based on a comparative proteomic analysis of these 2 types of LROs, we present a comprehensive AC proteome and identified 113 putative AC proteins. The methods and protein inventories provide a framework for studying the biogenesis and modification of LROs and the mechanisms by which they participate in regulating metal ion metabolism.

Spatiotemporal characteristics of P-selectin-induced ß2 integrin activation of human neutrophils under flow.

Activation of integrins is crucial for recruitment of flowing leukocytes to inflammatory or injured vascular sites, but their spatiotemporal characteristics are incompletely understood. We discovered that ß2-integrin activation over the entire surface of neutrophils on immobilized P-selectin occurred via mitogen-activated protein kinase (MAPK) or non-MAPK signaling with a minute-level timescale in a force-dependent manner. In flow, MAPK signaling required intracellular Ca2+ release to activate integrin within 2 min. Integrin activation via non-MAPK signaling occurred first locally in the vicinity of ligated P-selectin glycoprotein ligand-1 (PSGL-1) within sub-seconds, and then over the entire cell surface within 1 min in an extracellular Ca2+ influx-dependent manner. The transition from a local (but rapid) to global (but slow) activation mode was triggered by ligating the freshly activated integrin. Lipid rafts, moesin, actin, and talin were involved in non-MAPK signaling. Fluid loads had a slight effect on local integrin activation with a second-level timescale, but served as enhancers of global integrin activation.

[The characteristics of neutrophil extracellular traps produced by all-trans retinoic acid-induced dHL-60 under PMA stimulation].

Extracellular traps released by neutrophils (neutrophil extracellular traps, NETs) are a double-edged sword, and understanding the mechanism of NET formation is of great significance for disease treatment. However, the short lifespan, the large individual differences, and the inability to perform gene editing render it difficult to decipher NET formation using neutrophils. It is necessary to find a model cell to replace neutrophils to study the mechanism of NET formation. In this study, we used different concentrations (0, 0.1, 1, and 10 µmol/L) of all-trans retinoic acid (ATRA) to differentiate HL-60 cells for different days (1, 3, 5, and 7 days). By detecting the cell viability and nuclear morphology of cells, we confirmed that HL-60 cells were differentiated to neutrophil-like cells (dHL-60) after treated with ATRA for at least 5 days. Using immunofluorescence staining to detect the formation of NETs, we demonstrated that dHL-60 cells differentiated for 5 days with 1 µmol/L ATRA could generate NETs comparable to those produced by neutrophils upon phorbol 12-myristate 13-acetate (PMA) stimulation, without histone H3 citrullination. Furthermore, the formation of NETs by dHL-60 cells were NADPH-dependent and PAD4-independent, consistent with neutrophils. Taken together, these observations suggest that dHL-60 cells differentiated with 1 µmol/L ATRA for 5 days can be used as a model cell for neutrophils to study the mechanism of NET formation.

Shear Stress Accumulation Enhances von Willebrand Factor-Induced Platelet P-Selectin Translocation in a PI3K/Akt Pathway-Dependent Manner.

Platelet adhesion and activation through the interaction of von Willebrand factor (VWF) with platelet glycoprotein (GP) Ibα are the early key events in hemostasis and thrombosis especially under high blood shear stress. P-selectin translocation from α granule to the cell surface is a typical platelet function phenotype, which makes the platelet-induced inflammatory response of flowing leukocytes possible and can be induced by either chemical agonists (thrombin, ADP, etc.) or high blood shear stress, but regulations of VWF mutation and blood shear stress on VWF-induced P-selectin translocation remain unclear. With flow cytometry, parallel plate flow chamber, and immunofluorescence staining techniques, we examined the P-selectin translocation of platelets on immobilized wild-type (WT) VWF-A1 domain and its two mutants, the gain-of-function (GOF) mutant R1308L and the loss-of-function (LOF) mutant G1324S, respectively. The results showed that the VWF-A1-induced platelet P-selectin translocation was triggered, accelerated, and enhanced by fluid shear stress and could be correlated with shear stress accumulation (SSA, the product of fluid shear stress and mechanical stimulus time), and the PI3K/Akt axis was involved in the platelet P-selectin translocation. The force-triggered P-selectin translocation occurred quickly on partial platelet surface first and then extended gradually to the whole platelet surface as SSA increased. The P-selectin translocation process would be promoted by the GOF mutation (R1308L) but slowed down by the LOF mutation (G1324S). These findings demonstrated a force-enhanced regulation mechanism for the VWF-induced platelet P-selectin translocation through the PI3K/Akt pathway and provided a novel insight into the mechano-chemical regulation mechanism for the key events, such as platelet activation and functional phenotype change in hemostasis and thrombosis.

AFM Imaging Reveals Multiple Conformational States of ADAMTS13.

BACKGROUND: ADAMTS13 (A disintegrin and metalloprotease with a thrombospondin type 1 motif 13) cleaves Von Willebrand factor (VWF) to regulate its size, thereby preventing aberrant platelet aggregation and thrombus. Deficiency of ADAMTS13 caused by either genetic mutations or by inhibitory autoantibodies against ADAMTS13 leads to thrombotic thrombocytopenic purpura (TTP). Recently, ADAMTS13 was reported to adopt a "closed" conformation with lower activity and an "open" one resulting from the engagements of VWF D4-CK domains or antibodies to the distal domains of ADAMTS13, or mutations in its spacer domain. These engagements or mutations increase ADAMTS13 activity by ~ 2.5-fold. However, it is less known whether the conformation of ADAMTS13 is dynamic or stable. RESULTS: Wild type ADAMTS13 (WT-ADAMTS13) and the gain-of-function variant (GOF-ADAMTS13) with five mutations (R568K / F592Y / R660K / Y661F / Y665F) in spacer domain were imaged by atomic force microscopy (AFM) at pH 6 and pH 7.5. The data revealed that at both pH 6 and pH 7.5, WT-ADAMTS13 adopted two distinct conformational states (state I and state II), while an additional state (state III) was observed in GOF-ADAMTS13. In the present study, we propose that state I is the "closed" conformation, state III is the "open" one, and state II is an intermediate one. Comparing to pH 7.5, the percentages of state II of WT-ADAMTS13 and state III of GOF-ADAMTS13 increased at pH 6, with the decrease in the state I for WT-ADAMTS13 and state I and state II for GOF-ADAMTS13, suggesting lower pH extended the conformation of ADAMTS13. CONCLUSION: Both WT- and GOF-ADAMTS13 exist multiple conformational states and lower pH might alter the tertiary structure and/or disrupt the intra-domain interactions, increasing the flexibility of ADAMTS13 molecules.

Effect of ERK inhibitor on pulmonary metastasis of inoculated human adenoid cystic carcinoma cells in nude mice.

In this study, we detected the expression of phosphylated ERK1/2 in human salivary adenoid cystic carcinoma (SACC) by immunohistochemical method, the effect of various concentrations of PD98059 on the growth of ACC-M cells (a cell line of human adenoid cystic carcinoma of the salivary glands) by MTT assay, and the effect of PD98059 on the lung metastasis of inoculated ACC-M cells in nude mice. The results showed that the phosphylated ERK1/2 was overexpressed in SACC, the ACC-M cell line proliferation was inhibited by PD98059, and the lung metastasis level was reduced by PD98059, indicating that ERK may be a key molecule in targeted therapy of adenoid cystic carcinoma.

[Branchial cleft carcinoma from cleft cyst: report of one case].

Branchial cleft carcinoma is a rare malignancy, there still exist some controversies regarding to the differentiation of branchial cleft carcinomas. This article is aimed to familiarize clinicians with the presentation and treatment of the tumour and explores its origin. A 41-year old man who was seen with a large lesion of branchial cleft carcinoma with a slowly growing mass in the area of the left parotid gland was reported. The histological finding of dysplastic epithelium was next to direct invasive carcinoma. The diagnosis of a branchial cleft carcinoma requires the fulfillment of strict criteria. This case supports the origin of the carcinoma as being from an epithelial-lined cyst.