Introduction: Metabolic health status plays a central role in the development of a broad spectrum of chronic diseases. Yet existing assessment approaches rely on comparatively complex measurement techniques, which constrain their widespread adoption and limits their utility in population-based metabolic health management. Methods: The research employed a two-phase approach to construct a metabolic health index. We conducted a Delphi consultation to define and prioritize key metabolic health indicators, then drew on data from the China Chronic Disease and Risk Factor Surveillance, linked to the national mortality surveillance system, and built Cox proportional hazards models to estimate indicator-specific weights. Using these weights, we developed the Metabolic Health Index (MHI) and established a classification system to stratify metabolic health subtypes. We subsequently analyzed MHI score distributions and subtype prevalence across population subgroups. Results: The Delphi process identified 11 indicators for inclusion in the model: age, smoking, alcohol consumption, body-mass index, waist-to-height ratio, fasting blood glucose, blood pressure, blood lipids, serum uric acid, metabolic-associated fatty liver disease, and family history of metabolic diseases. Applying the classification system, we identified 13 metabolic health subtypes across 7 categories. MHI scores declined with age, and females consistently outscored males across all age groups. Over half of the participants (54.5%) had relatively high MHI scores, and the proportion with higher scores decreased with age. Conclusion: The MHI converts population health data into a quantitative metric for assessing metabolic health in adults and provides a practical approach for public health practice across diverse regions and settings.

IntroductionSepsis remains a leading cause of mortality in critical care, with dysregulated inflammatory responses driving disease progression. However, the role of necroptosis in sepsis pathogenesis remains incompletely understood.MethodsHere, through integration of multi-center cohort data (n = 1,265) and weighted gene co-expression network analysis (WGCNA), we constructed a six-gene necroptosis signature (CEBPD, CEBPB, MARCKS, SOCS3, PIM3, and JUNB) that correlated with sepsis severity and outcomes. Subsequently, we detected the expression of these genes in whole blood using qPCR. Furthermore, machine learning models incorporating this signature were evaluated across independent cohorts. Single-cell data analysis and flow cytometric analysis were further performed to characterize CD177+ neutrophils in sepsis.ResultsAll six Model-score genes were upregulated in sepsis patients, with four of them showing significant differences. Machine learning models incorporating this signature achieved robust diagnostic performance across independent cohorts. At the transcriptomic level, necroptosis activation showed a strong correlation with both the IL-6/STAT3 and TNF-α/NF-κB inflammatory pathways and distinct myeloid subsets. Single-cell data analysis further revealed that CD177+ neutrophils were significantly enriched in non-surviving sepsis patients and exhibited the highest necroptosis transcriptional score. Flow cytometric analysis revealed a significant increase in neutrophils, particularly the CD177+ subset, in the whole blood of septic patients. Furthermore, CD177+ neutrophils displayed blunted interferon responses alongside heightened production of inflammatory mediators, with nitric oxide (NO) potentially serving as an associated factor.DiscussionCollectively, our findings suggest that CD177+ neutrophils may be involved in necroptosis-related inflammation in sepsis and provide a clinically relevant gene signature for patient stratification, offering new perspectives for potential therapeutic exploration in sepsis management.

Background:Sepsis-associated acute kidney injury (SA-AKI) is a common and high-mortality complication in the intensive care unit (ICU), and renal replacement therapy (RRT) is frequently used, yet its survival benefits remain unclear. This study aims to assess survival differences via restricted mean survival time (RMST) in patients with SA-AKI. Methods:This retrospective cohort study included 577 patients diagnosed with SA-AKI from the ICU of Shenzhen People's Hospital between January 2017 and July 2019. All patients were categorized into RRT group (n=343) and non-RRT group (n=234) according to whether they need to received RRT. Survival analyses were conducted using Kaplan-Meier curves. Univariate and multivariable logistic regression analyses were employed for the selection of covariates. RMST analysis was performed to quantify the survival benefits associated with RRT. Results:Patients who need to received RRT had greater baseline illness severity and more frequent organ support (eg, higher SOFA scores and higher rates of shock and mechanical ventilation), suggesting potential confounding by indication. In unadjusted analyses, 28d mortality was higher in the RRT group than in the non-RRT group (44% vs 20%). After adjustment for multidrug-resistant (MDR) infection and SOFA score (both P < 0.05), the RMST difference between the RRT and non-RRT groups was 3.25 days (95% CI: 0.67 to 5.84; p = 0.01), which was statistically significant. Conclusion:In this retrospective cohort study, after adjusting for SOFA score and MDR infection, the potential survival benefits of RRT in patients with SA-AKI were demonstrated. These findings suggest that poor outcomes in RRT recipients may be attributable to underlying risk factors rather than RRT itself. However, given the observational design, these results are hypothesis-generating and require prospective validation.

Traditionally, the AJCC TNM staging system has been the primary tool for assessing the severity and prognosis of laryngeal cancer. Although several studies have demonstrated that the log odds of positive lymph nodes (LODDS) offers superior predictive accuracy compared to the TNM staging for other cancers, there is limited research for laryngeal cancer. This study analyzed data from SEER database (2000-2019). Independent risk factors for survival were identified using univariate and multivariate Cox regression analyses, and different prognostic models were constructed based on the multivariate analysis results. The predictive performance of these models was evaluated using receiver operating characteristic (ROC) curves and area under the curve (AUC) values. The results indicated that LODDS subgroup, age, marital status, histologic grade, T-stage, and N-stage were consistent independent prognostic factors for overall survival (OS) and cancer-specific survival (CSS). Assessment metrics showed that the multivariate model, which incorporated both LODDS and N staging, outperformed the individual N staging and LODDS models in predicting postoperative prognosis in laryngeal cancer patients. Overall, the multivariate model constructed in this study is a superior tool for predicting the postoperative status of laryngeal cancer.

Aim: Identifying the common functional single-nucleotide polymorphisms (SNPs) that can both affect the susceptibility to idiopathic pulmonary fibrosis (IPF) and silicosis. Methods: We first integrated the genome-wide association studies (GWASs) of IPF and silicosis to obtain the shared SNPs. Following this, functional expression quantitative trait locus (eQTL)-SNPs were identified by the GTEx database. This was followed by the validation of the correlation between these eQTL-SNPs and silicosis susceptibility through an additional case-control study including 194 silicosis cases and 235 healthy controls. Results: A total of 10 eQTL-SNPs that may affect silicosis susceptibility (P < 0.05) were obtained after the integration of the GWASs of IPF and silicosis, and a series of rigorous selection principles. Subsequently, the results of integrating the validation stage and the screening stage indicated that the variant T allele of rs1620530 located in the MAD1L1 (additive model: OR= 1.56, 95 % CI = 1.21-2.01, P = 0.001) and the variant G allele of rs2070063 located in the SERTAD2 (additive model: OR= 1.60, 95 % CI = 1.24-2.06, P < 0.001) were associated with increased silicosis susceptibility. The joint analysis indicated the risk of developing silicosis was higher in individuals who carried more unfavorable alleles of rs1620530 and rs2070063. Conclusions: The rs1620530 and rs2070063 may affect the silicosis susceptibility by regulating the expression of the MAD1L1 and SERTAD2, respectively. Further biological experiments are warranted to elucidate the underlying biological mechanisms between these two SNPs and the increased susceptibility to silicosis.

Thiols have interesting bio-chemical properties and can be found in a number of approved drugs. However, some thiols exhibited poor plasma stability and microsomal stability, leading to poor in vivo activity and poor oral bio-availability, in spite of their potent activity in vitro. Prodrug is a classic strategy to improve drug pharmacokinetics. In this study, we designed and synthesized 25 prodrug derivatives of a potent thiol-based HDAC inhibitor, IYS-15, to explore the structure-plasma stability relationships and structure-microsomal stability relationships in these series. We also tried to identify the main metabolic enzymes participated in the metabolism of some representative thiol-based prodrug derivatives. This work thus presents a comparison between different prodrugs based on the same model thiol, giving insights into the stability profile of the synthesized prodrug derivatives in human plasma and human liver microsomes, and more importantly, might also provide structural guidance to medicinal chemists in the design of thiol-based prodrugs and other novel prodrugs with thiol-based linkers.

Smoking is one of the most recognized risk factors for pulmonary fibrosis (PF). However, the underlying mechanism is not well understood. This study reveals smoking increases the risk of developing idiopathic PF (IPF) and that smoked IPF patients exhibit higher levels of senescence markers than non-smoker IPF patients. Moreover, smoking enhances bleomycin (Bleo)-induced PF, along with obvious senescence of type II alveolar (AT2) cells. RNA-seq assay identifies cigarette downregulates protection of telomeres 1 (POT1), which is then validated to decrease in smoked PF patients and mice via upregulating the methyltransferase MECP2. Mechanistically, MECP2 binds to the DNA methyltransferases (DNMTs)-induced methylated CpG island in the POT1 promoter, and smoking inhibits the transcriptional activity of the CpG island. The transcription factor FOXP2 could bind to this CpG island to promote POT1 transcription. However, this process is inhibited by forming a MECP2-FOXP2 complex, which blunts the FOXP2-POT1 DNA binding. siRNA-mediated POT1 knockdown promoted AT2 cell senescence in a p-ATM and p-ATR-dependent manner and secreted inflammatory and profibrotic factors, further promoting fibrotic response in fibroblasts. In vivo, delivery of the adeno-associated virus 9-POT1 (AAV9-POT1) vector inhibits cigarette-induced cell senescence and effectively alleviates PF in mice. These findings demonstrate that POT1 is an essential protector in PF by protecting against AT2 cell senescence.

BACKGROUND:This study aimed to evaluate the safety and efficacy of Venastent for the patients with iliac vein compression syndrome (IVCS). METHODS:A multicenter retrospective clinical trial enrolling 199 IVCS patients was conducted by the department of vascular surgery of 3 hospitals from October 2021 to January 2023. Among them, 91 were treated with a new type of iliac vein stent and defined as the experimental group, and 108 cases in the control group were using Zilver Vena venous stent (Cook Medical). Twelve-month follow-up was performed for the following primary end points: the patency rate, the venous clinical severity score (VCSS), and the Clinical, Etiology, Anatomy, and Pathology (CEAP) classification. RESULTS:To validate the role of Venastent in the prevention of thrombosis, 91 patients with Venastent versus 108 patients with control stent were compared. No difference was found in the demographic data or accompanying symptoms or disease severity scores between the 2 groups of patients. The patency rates of the contralateral deep vein at 24 months after the surgery were 100.00% vs. 94.5%, respectively (P = 0.043). Moreover, no difference was found on the VCSS score and CEAP classification of the both groups. CONCLUSION:Venastent with a large corolla-like mesh and an adequate radial force shows a good patency in the ipsilateral iliac vein and could prevent thrombosis in the contralateral iliac vein.

Cadmium (Cd) is a highly toxic metal in human body, and therefore understanding the immunotoxicity of Cd is significant for public health. The aim of this study was to investigate the role of hematopoietic stem cells (HSC) in regulating the immunotoxicity of Cd. After exposure to 10 ppm Cd via drinking water for up to 9 months, C57BL/ 6 mice had a suppressed adaptive immune system at day 135 but had an enhanced adaptive immune system at day 270 during Cd exposure. The biphasic impacts of Cd on the adaptive immune system were correlated to the mitochondrial (MT) activation of HSC. Mechanistically, a direct action of Cd activated the non-canonical Wnt signaling to increase MT activation in HSC in the bone marrow (BM) at day 90, thus resulting in an impaired adaptive immune system in mice at day 135 during Cd exposure; conversely, Cd reduced the production of thrombopoietin (TPO) by osteoblasts in the BM to suppress MT activation in HSC at day 180, which in turn caused an enhanced adaptive immune system in mice at day 270 during Cd exposure. Thus, Cd biphasically impacts the adaptive immune system via regulating MT activation of HSC, providing a novel angle for understanding the immunotoxicology of metals.

Subcision is one of the most frequently used surgical options in improving acne scarring. Concentrated growth factors (CGF), with the potential to assist in the repair and regeneration of tissues, have been extended to the field of cosmetic plastic surgery. Here, we investigate the safety and efficacy of combining CGF (the latest generation of platelet concentrates) with subcision as a novel treatment method. Sixty patients with clinically diagnosed acne scars received either subcision combined with CGF (n = 24) or subcision monotherapy (n = 36). Clinical characteristics and treatment outcomes were documented through photographs, questionnaires, and medical records, with side effects recorded. Both groups showed significant scar improvements, but the combination therapy demonstrated superior outcomes in ECCA score reduction compared to subcision treatment alone. Patients in the combination therapy group reported higher satisfaction levels, and side effects were minimal, except for increased ecchymosis and pain levels which resolved within an acceptable recovery time range for the patients. This combination approach efficiently treats severe acne scars without significant safety risks.

Alzheimer’s disease (AD) and ischemic stroke (IS) are prevalent neurological disorders that frequently co-occur in the same individuals. Recent studies have demonstrated that AD and IS share several common risk factors and pathogenic elements, including an overlapping genomic architecture. However, the relationship between IS risk gene polymorphisms and AD has been less extensively studied. We aimed at determining whether IS risk gene polymorphisms were associated with the risk of AD and the severity of AD in AD patients. We utilized data of AD patients and normal controls (NCs) sourced from the Alzheimer’s Disease Neuroimaging Initiative (ADNI) cohort. IS risk single nucleotide polymorphisms (SNPs) were identified through the most recent and largest IS genome-wide association study (GWAS) meta-analysis. Subsequently, we conducted SNP-based association analysis of IS-risk SNPs with the risk of AD, along with amyloid, tau, and neuroimaging for AD. The generalized multifactor dimensionality reduction (GMDR) model was used to assess the interactions among IS-risk SNPs and apolipoprotein E (ApoE) ε4. Protein–protein interactions (PPIs) of the IS-risk genes product and APOE were explored using the STRING database. Seven IS-risk SNPs were involved in the study. Five SNPs were found to be associated with at least one measurement of cerebrospinal fluid (CSF) levels of amyloid-beta 1–42 (Aβ42), total tau (t-tau), and phosphorylated tau 181 (p-tau181), as well as the volumes of the hippocampus, whole brain, entorhinal cortex, and mid-temporal regions. After multiple testing corrections, we found that T allele of rs1487504 contributed to an increased risk of AD in non-ApoE ε4 carriers. The combination of rs1487504 and ApoE ε4 emerged as the optimal two-factor model, and its interaction was significantly related to the risk of AD. Additionally, C allele of rs880315 was significantly associated with elevated levels of CSF Aβ42 in AD patients, and A allele of rs10774625 was significantly related to a reduction in the volume of the entorhinal cortex in AD patients. This study found that IS risk SNPs were associated with both the risk of AD and AD major indicators in the ADNI cohort. These findings elucidated the role of IS in AD from a genetic perspective and provided an innovative approach to predict AD through IS-risk SNPs.

Keloid is a typical skin fibrotic disease with unclear mechanisms and limited therapeutic options. Fibroblast-induced fibrogenesis is a crucial cause of KD. However, the types of cells involved in fibroblast fibrogenesis in KD and the specific mechanisms are unclear. This study aimed to investigate the role of melanocyte-secreted melanin in promoting fibroblast fibrogenesis and its mechanism and to evaluate the potential therapeutic effect of intervening melanin in treating keloid. The activity of pigmentation-related pathways in KD melanocytes was examined using single-cell RNA-sequence (scRNA-seq) analysis. Masson-Fontana staining or isolated melanin quantification detected the melanin levels and distribution in the skin and cells. Collagen deposition, wounding healing, and proliferation analysis were employed to integratively assess fibroblast fibrogenesis. After melanin treatment, bulk-seq identified fibroblasts’ differentially expressed genes (DEGs). The iron levels were detected by Perl’s staining or isolated iron quantification. Cell viability, LipidROS, and malondialdehyde assay accessed the ferroptosis levels. The therapeutic potential of ML329 was evaluated in keloid-bearing mice. We found the enriched skin pigmentation-related pathways in the melanocytes of keloid by single-cell RNA-sequence (scRNA-seq) analysis. We further validated increased melanin levels in keloid patients. Additionally, melanin positively correlated with the Keloid Area and Severity Index in keloid. Furthermore, melanocyte-secreted melanin significantly promoted fibroblast proliferation, migration, and collagen synthesis. Mechanically, melanin increased basal cell permeability and inflammation to facilitate its transfer to the dermis, where it further activated fibroblasts by evoking iron overload and ferroptosis resistance. Consistently, iron overload and ferroptosis resistance were validated in primary fibroblasts and skin tissues of keloid patients. Inhibition of iron overload and ferroptosis resistance effectively diminish melanin-induced fibrogenesis. Interestingly, melanin induced iron overload and ferroptosis resistance in melanocytes in an autocrine manner and further stimulated keratinocytes to take up melanin to deepen skin color by upregulating the F2R-like trypsin receptor 1 (F2RL1). In vivo, the delivery of ML329, a microphthalmia-associated transcription factor (MITF) inhibitor, could suppress melanogenesis and alleviate keloid in human keloid-bearing nude mice. Meanwhile, ML329 decreased the iron content and restored the sensitivities of ferroptosis. Collectively, melanin-lowing strategies may appear as a potential new therapeutic target for keloid. Current treatments for keloid are ineffective. Our research demonstrates that melanin levels increase in keloid patients and play a significant role in keloid progression by mediating aberrant keratinocyte − melanocyte − basal − fibroblast cell crosstalk. Importantly, we found that pharmacological inhibition of melanogenesis using an MITF inhibitor, ML329, shows promise in effectively alleviating keloid, offering a potential breakthrough in keloid treatment. The melanin synthesis pathway is abnormally activated in KD melanocytes. Melanin destroys the basal membrane barrier by triggering inflammation and translocates to the dermal layers of the skin in a paracrine manner to induce fibroblast overgrowth, migration, and ECM deposition by inducing iron overload and ferroptosis resistance. Melanin maintains melanocytes’ hyperproliferative and non-immortal properties in an autocrine manner. It further enhances the keratocyte PAR-2 to promote transit to the superficial epidermis skin layers, which may be related to the deepening of skin color. Inhibition of melanin synthesis by ML329 alleviates KD in mice.

Abstract Background Observational studies have elucidated the associations between dietary factors and hypertension. Nevertheless, the exploration of these relationships using Mendelian randomization remains scarce currently. Methods The Mendelian randomization approach investigated the potential causal relationships between 16 dietary factors and hypertension. To achieve this, we identified genetic variants associated with these dietary factors by utilizing data from European-descent genome-wide association studies with a stringent significance threshold (P < 5 × 10 − 8). Subsequently, we obtained genetic associations with hypertension from the extensive FinnGen Study, encompassing 92,462 cases and 265,626 controls. Our primary analytical method was the inverse variance weighted method, and we also conducted assessments for heterogeneity and pleiotropy to ensure the robustness and reliability of our findings. Results The study revealed significant associations with hypertension risk for various dietary factors. Specifically, higher weekly alcohol consumption (OR: 1.53, 95% CI: 1.19–1.96) and more frequent alcohol intake (OR: 1.20, 95% CI: 1.08–1.33) were positively correlated with an increased risk of hypertension. Likewise, increased poultry intake (OR: 3.25, 95% CI: 1.83–5.78) and beef intake (OR: 1.80, 95% CI: 1.09–2.97) were also linked to a higher risk of hypertension. Conversely, there were protective factors associated with a decreased risk of hypertension. These included consuming salad and raw vegetables, dried fruits, cheese, and cereals. It is important to note that no evidence of pleiotropy was detected, underscoring the robustness of these findings. Conclusions This study uncovered causal relationships between various dietary factors and hypertension risk. Specifically, alcohol consumption in terms of drinks per week and intake frequency, as well as poultry and beef intake, were causally associated with an elevated risk of hypertension. In contrast, consuming salad/raw vegetables, dried fruits, cheese, and cereals demonstrated an inverse causal association with hypertension, suggesting a potential protective effect.

Background:Several large-scale observational studies have found deep vein thrombosis (DVT) to be related with coronavirus disease 2019 (COVID-19). However, whether there is a clear causal connection between the two is unknown. Methods:Our primary analytical method was the inverse variance-weighted (IVW) approach, complemented by the Mendelian randomisation-Egger (MR-Egger) and weighted median methods. We also used MR-Egger to examine the presence of pleiotropy and the Mendelian randomisation pleiotropy residual sum and outlier (MR-PRESSO) approach to analyse for heterogeneity in the data. Results:We did not observe a direct causal relationship between COVID-19 susceptibility (odds ratio (OR) = 1.023; 95% confidence interval (CI) = 0.828-1.264, standard error (SE) = 0.108, P = 0.833), hospitalisation (OR = 1.030; 95% CI = 0.943-1.125, SE = 0.374, P = 0.720), severity (OR = 0.994; 95% CI = 0.923-1.071, SE = 0.038, P = 0.877), and DVT. The results of the reverse Mendelian randomisation (MR) for DVT and COVID-19 susceptibility exhibited heterogeneity and horizontal pleiotropy. Even after removing outliers, we detected no direct causal relationship between the two (OR = 1.015; 95% CI = 0.954-1.080, SE = 0.032, P = 0.630). Similarly, we found no direct causal relationship between DVT and COVID-19 hospitalisation (OR = 0.999; 95% CI = 0.907-1.102, SE = 0.050, P = 0.999) or severity (OR = 1.014; 95% CI = 0.893-1.153, SE = 0.065, P = 0.826). Conclusions:In this MR study, we identified no direct causal impact in a European population between DVT and the COVID-19 susceptibility, severity, or hospitalisation.

Backgrounds and Aims: Atrial fibrillation (AF) is the most common cardiac arrhythmia among the older patients (>= 80 years) in clinical practice. The index of neutrophil percentage-to-albumin ratio (NPAR) is a reliable predictor of adverse outcomes in cardiovascular diseases. There is scarce evidence regarding the association between NPAR and mortality among the older patients with AF. Methods: The research was conducted among 1141 patients with AF between January 2015 and June 2020, hospitalized at Huadong Hospital affiliated with Fudan University. The primary outcome were 28-day all-cause and cardiovascular mortality. Cox regression analysis and Kaplan-Meier survival curves were used to explore the correlation between NPAR and 28-day all-cause or cardiovascular mortality. Receiver operating characteristic (ROC) curve and the area under the curve (AUC) were performed for the predictive values of NPAR on prognosis. Results: The 28-day death rate from cardiovascular disease and all-causes were 3.3% and 8.7%, respectively. Continuous NPAR levels were positively associated with all-cause (HR 1.13, 95% CI 1.09, 1.16) and cardiovascular (HR 1.16, 95% CI 1.10, 1.23) mortality after adjustment for confounding variables. Relative to patients in the T1 group, those in higher NPAR tertiles also exhibit elevated risks of all-cause (P < 0.001) and cardiovascular mortality (P < 0.001). Furthermore, both all-cause and cardiovascular mortality rates rose with increasing NPAR in all analyzed subgroups. Conclusion: NPAR values are consistently positively related to 28-day all-cause and cardiovascular mortality rates in patients >= 80 years of age with AF.

Introduction: Atrial fibrillation (AF) is a prevalent heart arrhythmia in elderly adults aged 80 years or older. The red cell distribution width (RDW) to albumin ratio has been acknowledged as a reliable prognostic marker for poor outcomes in a variety of disorders. However, there exists limited scientific evidence on the association of RDW to albumin (RAR) with mortality in geriatric individuals with AF. Methods: From January 2015 to June 2020, a retrospective study was conducted in a tertiary academic institution that diagnosed 1,141 elderly adults with AF. The RAR value was calculated as the ratio of RDW (%) to albumin (g/dL). The potential association between RAR and cardiovascular mortality and the risk of all-cause mortality within 28 days was evaluated by means of multivariable Cox regression analysis. Results: The 28-day all-cause and cardiovascular mortality rates were 8.7% and 3.3%, respectively. Increased RAR tertiles were found to be significantly associated with greater all-cause mortality (T1: 1.6%; T2: 6.2%; T3: 18.1%, p < 0.001) and cardiovascular mortality (T1: 0.8%; T2: 2.9%; T3: 6.3%, p < 0.001) using Kaplan-Meier analysis. Continuous RAR had a positive association with all-cause mortality (hazard ratios [HR] = 1.42, 95% confidence interval [CI] 1.23–1.65) and cardiovascular mortality (HR = 1.31, 95% CI: 1.05–1.64), even after accounting for numerous confounding variables. In comparison to the T1 group, individuals with the highest RAR levels displayed a greater risk of all-cause mortality (HR = 2.73, 95% CI: 1.11–6.74) and cardiovascular mortality (HR = 2.59, 95% CI: 0.69–9.78). Increased RAR levels were related to higher rates of cardiovascular and all-cause mortality across almost all subgroups. Conclusion: RAR is independently correlated with 28-day all-cause mortality and cardiovascular mortality in AF-affected individuals aged ≥80.

In this study, we developed and validated a simple, fast and sensitive LC-MS/MS method for the measurement of tetrodotoxin (TTX) in human plasma. Three HILIC-type solid phase extraction (SPE) carriers (PSA, silica, Siphila i HILIX) with different stationary phase functional groups were compared. The Siphila i HILIX SPE plate containing multi-carboxyl groups was finally selected due to obviously better extraction recovery of TTX (about 80% of recovery from plasma samples) than the other two and no significant matrix effects were observed, which was speculated to have mixed-mode synergistic effects of hydrophilic interaction and ion exchange. 100μL plasma sample was precipitated rapidly with acetonitrile containing 1% trichloroacetic acid, and filtrates were loaded onto Siphila i HILIX 96 well SPE plate. After washed with 95% acetonitrile, TTX was eluted with 200μL of 50% acetonitrile containing 1% trichloroacetic acid. 2μL of elution solution was directly injected into LC-MS/MS and the total run time on a BEH amide column was 4.5 min. The method avoids the evaporation and ultrafiltration processes which is simple and timesaving (<30 min). TTX and internal standard (arginine-15N4) were monitored in positive mode using m/z 320.3→162.2 (quantification transition for TTX), 320.3→284.1 (confirmation transition for TTX) and 179.2→63.0 (transition for IS), respectively. The method was linear in the range of 0.1-20 ng/mL for TTX with the low limit of quantification (S/N > 10) of 0.1 ng/mL; the intra- and inter-assay accuracies were in the range of 98.5%-99.8% (relative standard deviations, RSDs ≤ 5.92%) and 98.8-99.5% (RSDs ≤ 6.23%), respectively. Biases of spiking analysis were ranged from -7.00% to 7.43% for healthy human plasma samples (RSDs ≤ 8.83%) and from -5.00% to 3.93% for hemolytic, high triglyceride, high cholesterol and high bilirubin plasma samples (RSDs ≤ 6.40%), which proved the good anti-interference property of the method. The results showed that the method is sensitive, accurate, specific, reliable, and can be used to monitor the concentration of TTX in plasma to meet the needs of clinical research and poisoning screening.

We aimed to investigate the effects of CX-C chemokine receptor type 4 (CXCR4) on transforming growth factor (TGF)-β1-induced cardiac fibrosis in Human cardiac fibroblasts (HCFs). HCFs were stimulated with TGF-β1, and the level of α-smooth muscle actin (α-SMA) was assessed by immunofluorescence assay. The expression of CXCR4 was detected by Western blotting. Then the cells were incubated with CXCR4 antagonist AMD 3465. Cell viability was measured by CCK-8 assay. The expression of α-SMA, proliferating cell nuclear antigen (PCNA) and Ki67 were examined. Collagen synthesis was detected by sirius red staining. Moreover, the expression of phpspho-Smad2 (p-Smad2) and p-Smad3 were determined. We found that the level of α-SMA was increased after induction with TGF-β1. The expression of CXCR4 was upregulated in TGF-β1-treated HCFs. Following treatment with AMD 3465, cell proliferation was inhibited coupled with a decrease in PCNA and Ki67 expression. Additionally, the expression of α-SMA was decreased after being intervened with AMD 3465. Concurrently, the levels of collagen were reduced accompanied by downregulation of Collagen I and III. Furthermore, AMD 3465 treatment decreased the expression of p-Smad2 and p-Smad3. Our findings suggested that CXCR4 antagonist AMD 3465 could alleviate cardiac fibrosis via blocking TGF-β1-induced activation of Smad2/3 in HCFs.

Effect of atorvastatin inhibition of endoplasmic reticulum stress and amelioration of atherosclerosis through AMPK pathway were studied. Eight-week-old male apolipoprotein E-deficient (ApoE(-)/(-)) mice were fed with high-fat diet for 2 weeks and randomly divided into two groups: Atorvastatin treatment group was given atorvastatin (5 mg/kg/day) injection for a total of 6 weeks; control group was given the same dose of PBS through intraperitoneal injection for a total of 6 weeks. H&E staining was used to detect plaque size; immunohistochemical staining was used to detect T cells, macrophages and phospho-protein kinase-like ER kinase (phospho-PERK) in localized plaques. Proteins were extracted from mouse thoracic and abdominal aortic tissues. Western blot analysis was used to detect the protein expression levels of endoplasmic reticulum stress-related molecules phospho-eukaryotic initiation factor-2 alpha (p-eIF2 alpha), eukaryotic initiation factor (eIF2a), and sliced x-box binding protein 1 (sXBP-1). Cultured human umbilical vein endothelial cells (HUVECs), induced endoplasmic reticulum stress with human oxidized low density lipoprotein (ox-LDL), were treated with atorvastatin, AMPK agonist 5-amino-4-imidazolecarboxamide riboside-I-beta-D-ribofuranoside (AICAR) and AMPK-DN that expressed a dominant-negative mutant of AMPK. Western blot analysis was used to test the expression levels of endoplasmic reticulum stress-related molecules p-elF2a and sXBP-1. The area of aortic plaques in atorvastatin group was obviously decreased, and the infiltrations of CD3(+) T cells and macrophages in the localized plaques were reduced. The endoplasmic reticulum stress-related proteins sXBP-1 and p-eIF2a were significantly reduced. The results of immunohistochemistry also showed a significant decrease in the level of phospho-PERK (p-PERK) in atorvastatin group. The results in ox-LDL-induced HUVECs showed that atorvastatin inhibited ox-LDL-induced endoplasmic reticulum stress, and the AMPK agonist AICAR also had the same effect, which was offset by DN-AMPK treatment. Atorvastatin inhibits ER stress both in vitro and in vivo and this protective effect is mediated by AMPK activation.

An experimental study of the interaction between short wind-waves and internal wave is reported. It is shown that at low wind speed internal wave excited by moving object in stratified fluid changes the distribution of short wind-waves by changing surface current field. This phenomenon is well explained by a wave-current interaction mode. It implies that the wake of under-water moving object can be distinguished from the changes of the distribution of short wind-waves.