Real-time, random-access organ screening for carbapenem-resistant organisms (CRO) reduces CRO-associated, donor-derived infection mortality in lung transplant recipients.

PURPOSE: Donor-derived infection (DDI) has become an important factor affecting the prognosis of lung transplantation patients. The risks versus benefits of using donor organs infected with multidrug-resistant organisms (MDRO), especially carbapenem-resistant organisms (CRO), are frequently debated. Traditional microbial culture and antimicrobial susceptibility testing at present fail to meet the needs of quick CRO determination for donor lungs before acquisition. In this study, we explored a novel screening method by using Xpert® Carba-R assay for CRO in donor lungs in a real-time manner to reduce CRO-associated DDI mortality. METHODS: This study was registered on chictr.org.cn (ChiCTR2100053687) on November 2021. In the Xpert Carba-R screening group, donor lungs were screened for CRO infection by the Xpert Carba-R test on bronchoalveolar fluid (BALF) before acquisition. If the result was negative, donor lung acquisition and subsequent lung transplantation were performed. In the thirty-five potential donors, nine (25.71%) with positive Xpert Carba-R results in BALF were declined for lung transplantation. Twenty-six recipients and the matching CRO-negative donor lungs (74.29%) were included in the Xpert Carba-R screening group. In the control group, nineteen recipients underwent lung transplants without Xpert Carba-R screening. The incidence and mortality of CRO-associated DDI were collected and contrasted between the two groups. RESULTS: Multivariate analysis showed that CRO-related death due to DDI within 60 days was significantly lower in the Xpert Carba-R screening group than that in the control group (OR = 0.05, 95% CI 0.003-0.74, p = 0.03). CONCLUSION: Real-time CRO screening of donor lungs before transplantation at the point of care by the Xpert Carba-R helps clinicians formulate lung transplantation strategies quickly and reduces the risk of subsequent CRO infection improving the prognosis of lung transplantation.

Trichosanthin-derived peptide Tk-PQ attenuates immune rejection in mouse tracheal allotransplant model by suppressing PI3K-Akt and inducing type II immune polarization.

Obliterative bronchiolitis (OB) is one of the main complications affecting long-term survival of post-lung transplantation patients. In this study, we evaluated the efficacy of Tk-PQ (a peptide derived from trichosanthin) in alleviating OB in a mouse ectopic tracheal transplant model. We found that post-transplantation treatment of Tk-PQ significant ameliorated OB symptoms including luminal occlusion, epithelial cells loss and fibrosis in the allograft. In addition, Tk-PQ promoted immune suppressive environment by inducing Th2 polarization and increasing Treg population which in turn led to elevated levels of anti-inflammatory cytokines IL-4, IL-10, IL-33 and decreased levels of pro-inflammatory IL-1ß. Mechanistically, we used transcriptome analysis of splenic T cells from allografted mice to show that Tk-PQ treatment down-regulated the PI3K-Akt signaling pathway. Indeed, the immune suppression phenotypes of Tk-PQ was recapitulated by a PI3K inhibitor LY294002. Taken together, Tk-PQ regulates post-transplantation immuno-rejection by modulating the balance of T cell response via the PI3K-Akt pathway, making it a promising peptide based immune rejection suppressant for patients receiving allotransplant.

The molecular targets of Kangai injection in gastric cancer by in silico network pharmacology approach and experiment confirmation.

INTRODUCTION: This study aimed to identify the phytochemical constituents that could target gastric cancer in Kangai injection using a network pharmacology-based approach. METHODS: Protein-protein interactions (PPI), Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were conducted utilizing String and OmicShare tools. In the in vitro experiments, the related mRNA and protein levels were assessed via real-time quantitative polymerase chain reaction and Western blotting, respectively. Cell proliferation was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay. RESULTS: Kangai injection comprises several compounds, which target multiple substrates and pathways related to gastric cancer. The PPI and Gene Ontology analyses revealed that tumor necrosis factor (TNF) was a hub gene. KEGG pathway enrichment analysis indicated that the the TNF pathway was significantly enriched. Kangai injection decreased the mRNA levels of TNFR2, TRAF2, PI3K, AKT, and IκBα and inhibited the phosphorylation of PI3K, AKT, and IκBα phosphorylations. Kangai injection inhibited cell proliferation, while TNFR2 overexpression or treatment with the PI3K activator 740 Y-P partially restored it. CONCLUSION: Kangai injection operates through multiple targets and pathways in gastric cancer, with the TNFR2/PI3K/AKT/NF-κB pathway playing a crucial role in its mechanism against gastric cancer.

S-Scheme Porphyrin Covalent Organic Framework Heterojunction for Boosted Photoelectrochemical Immunoassays in Myocardial Infarction Diagnosis.

Cardiac troponin I (cTnI) is an extremely sensitive biomarker for early indication of acute myocardial infarction (AMI). However, it still remains a tough challenge for many newly developed cTnI biosensors to achieve superior sensing performance including high sensitivity, rapid detection, and resistance to interference in clinical serum samples. Herein, a novel photocathodic immunosensor toward cTnI sensing has been successfully developed by designing a unique S-scheme heterojunction based on the porphyrin-based covalent organic frameworks (p-COFs) and p-type silicon nanowire arrays (p-SiNWs). In the novel heterojunction, the p-SiNWs are employed as the photocathode platform to acquire a strong photocurrent response. The in situ-grown p-COFs can accelerate the spatial migration rate of charge carriers by forming proper band alignment with the p-SiNWs. The crystalline π-conjugated network of p-COFs with abundant amino groups also promotes the electron transfer and anti-cTnI immobilizing process. The developed photocathodic immunosensor demonstrates a broad detection range of 5 pg/mL-10 ng/mL and a low limit of detection (LOD) of 1.36 pg/mL in clinical serum samples. Besides, the PEC sensor owns several advantages including good stability and superior anti-interference ability. By comparing our results with that of the commercial ELISA method, the relative deviations range from 0.06 to 0.18% (n = 3), and the recovery rates range from 95.4 to 109.5%. This work displays a novel strategy to design efficient and stable PEC sensing platforms for cTnI detection in real-life serums and provides guidance in future clinical diagnosis.

Long-term follow up and comparison between conservative and interventional therapy in postoperative bronchopleural fistula-a cohort study.

Background: Bronchopleural fistula (BPF) is a relatively rare postoperative complication with high mortality. The management is tough and controversial. The aim of this study was to compare the short and long-term outcomes between the conservative and the interventional therapy in postoperative BPF. We also concluded our own strategy and experience of treatment in postoperative BPF. Methods: Postoperative BPF patients with malignancies, aged from 18 to 80 years old who had undergone thoracic surgery between June 2011 and June 2020, were included in this study and followed up from 20 months to 10 years. They were retrospectively reviewed and analyzed. Results: Ninety-two BPF patients were included in this study, 39 of whom underwent interventional treatment. Significant differences were found in the 28-day and the 90-day survival rates between the conservative and the interventional therapy (P=0.001, 43.40% vs. 76.92%; P=0.006, 35.85% vs. 66.67%). Simple conservative therapy was independently associated with 90-day mortality between the groups in postoperative BPF [P=0.002, hazards ratio (HR) =2.913, 95% confidence interval (CI): 1.480-5.731]. Conclusions: Postoperative BPF is notorious for its high mortality. Surgical and bronchoscopic interventions are recommendable in postoperative BPF as they guarantee better short and long-term outcomes compared with the conservative therapy.

Pharmacokinetic Evaluation of Tacrolimus in Chinese Adult Patients during the Early Stages Post-Lung Transplantation.

BACKGROUND: Although tacrolimus has been widely used in patients undergoing lung transplantation, few studies have reported the pharmacokinetics of tacrolimus in Chinese patients after lung transplantation. Thus, we aimed to investigate the pharmacokinetics and influential factors in this patient cohort in the early stage after lung transplantation. METHODS: We enrolled 14 adult lung transplant recipients who were treated with tacrolimus and then intensively collected blood samples within a 12-h dosing interval. The pharmacokinetic parameters of tacrolimus were calculated using non-compartmental analysis, and the influence of pathophysiological characteristics and CYP3A5*3 and CYP3A4*1G genotypes on the pharmacokinetics of tacrolimus was assessed. Using linear regression analysis, we investigated the correlation between tacrolimus concentration at different sampling points and measured the area under the time-concentration curve (AUC0-12h). RESULTS: Geometric mean of apparent clearance (CL/F) was 18.13 ± 1.65 L/h in non-CYP3A5*3/*3 carriers, five times higher than that in CYP3A5*3/*3 carriers (p < 0.001). Furthermore, the tacrolimus concentration 4 h after administration had the strongest correlation with AUC0-12h (R2 = 0.979). CONCLUSION: The pharmacokinetics of tacrolimus varied largely between patients during the early stage post-transplantation, which could be partially explained by CYP3A5*3 genetic polymorphisms.

A novel photoelectrochemical sensor based on SiNWs@PDA for efficient detection of myocardial infarction.

Based on the necessity and urgency of cardiac troponin I (cTnI) detection for the diagnosis of myocardial infarction, a novel unlabeled photoelectrochemical (PEC) immunosensor has been developed to detect cTnI rapidly and sensitively. Silicon nanowire arrays (SiNWs) were prepared via metal-assisted chemical etching. To improve the stability and sensitivity towards cTnI sensing, the surface of silicon nanowire arrays were coated with polydopamine by an in situ growth method. PDA was uniformly modified on the nanowire surface to provide a reliable active site for antibody binding. The linear dynamic range of the cTnI detection method was 0.005-10 ng mL-1, and the detection limit was 1.47 pg mL-1. The designed PEC immunosensor exhibited good sensitivity, selectivity, stability and reproducibility. The electrode enabled label-free detection and provided a new route to realize point-of-care testing of cTnI.

Extraction optimization and screening of antioxidant peptides from grass carp meat and synergistic-antagonistic effect.

Grass carp (Ctenopharyngodon idellus) is one of the three most cultivated freshwater fish around the world, but it is mainly consumed afresh, so only a small part of them are processed into salted fish or snack food. This research was performed to prepare and screen antioxidant peptides from grass carp muscle to promote its high-value utilization. The parameters of double-enzyme two-step hydrolysis were optimized, the peptides with the highest ABTS.+ scavenging ability were enriched and identified by Sephadex G-25 and LC-Q-Orbitrap-MS/MS. The synergistic-antagonistic effect among identified peptides was also investigated. The optimized conditions were hydrolyzed with protamex (10,000 U/g) at pH 8.0, 50°C for 3 h, followed by hydrolysis with alcalase (6,000 U/g) at pH 9.0, 50 °C for 2 h, and the protein-liquid ratio was 4%. The hydrolysates were further fractionated to obtain five fractions, in which fraction 3 (F3) exhibited the strongest ABTS.+ and O 2 · - scavenging ability with the IC50 values of 0.11 and 0.47 mg/ml, respectively. Twelve novel antioxidant peptides were identified, in which VAGW possessed the highest activity (139.77 µmol GSH/g). Significantly synergistic effects were observed on the two and three peptides' combination among VAGW, APPAMW, LFGY, FYYGK, and LLLYK, while the C-terminal tryptophan (Trp) played an important role in the synergism. This study found that grass carp muscle hydrolysates can be potential natural antioxidants in functional products. The synergistic effects among peptides may provide a perspective for the combined application of peptides.

Normothermic ex vivo lung perfusion outperforms conventional cold preservation in a deceased rat lung.

Background: Utilization rate of donor lungs in China is constrained by the inefficiency of conventional cold preservation (COLD) in lung transplantation. We assessed the performance of normothermic ex vivo lung perfusion (EVLP) in lung donors following cardiac death (DCD) with fresh EVLP perfusate. Methods: A total of 24 adult Sprague-Dawley (SD) rats were randomized into three groups (EVLP, COLD, and control groups) with 8 rats each. Fresh EVLP perfusate was prepared for lung perfusion. The pH, electrolyte concentration, perfusate penetration pressure during irrigation, physiological function, integrity of barrier function, pathological changes, and expressions of inflammation-related cytokines of donor lungs were examined. Results: The pH, electrolyte concentration, osmotic pressure of the perfusate were kept stable for 4 hours after EVLP was administered. Pulmonary arterial pressure (PAP), pulmonary venous pressure (PVP), pulmonary venous oxygenation index (PaO2/FiO2), and end carbon dioxide partial pressure (PCO2) in the EVLP group were all within acceptable limits. After 4 hours, the oxygenation index was significantly higher in the EVLP group compared with that of the COLD group (P=0.0032). The EVLP group had a decreased wet/dry weight ratio (P=0.0155) and Evans blue dye content (P=0.0075) compared to that of the COLD group. Pathological examination showed more obvious lung damage in the COLD group than in the EVLP group. However, there was no difference in protein expression of high mobility group box 1 (HMGB1) and toll-like receptor 4 (TLR4) between the EVLP group and the COLD group. Conclusions: Fresh lung perfusate could be used for EVLP to protect and repair donor lungs. Normothermic EVLP performed better than conventional cold preservation. EVLP might be powerful for lung quality maintenance and may have potential for anti-inflammation.

IL-13 Derived Type 2 Innate Lymphocytes Ameliorates Cardiomyocyte Apoptosis Through STAT3 Signaling Pathway.

The involvement of cardiomyopathy during sepsis means higher mortality and prolonged length of hospital stay. Many efforts have been made to alleviate the apoptosis of cardiomyocytes in sepsis. The huge potential of IL-13 in tissue repair has attracted increasing attention. In the present study, we used LPS-treated mice or primary cardiomyocytes as a sepsis model to explore the anti-apoptotic ability of IL-13. It was found that an increased level of exogenous IL-13 was beneficial to the recovery of heart function in sepsis, and this anti-apoptotic effect of IL-13 was probably through enhancing the phosphorylation of STAT3 Ser727. In addition, we identified that the heart protective effect of IL-13 was associated with type 2 innate lymphocytes (ILC2). All these findings may provide a potential promising treatment for sepsis-induced cardiomyopathy.

Engineered Biomimetic Nanoplatform Protects the Myocardium Against Ischemia/Reperfusion Injury by Inhibiting Pyroptosis.

Protection of cardiomyocytes against oxidative stress is vital to alleviate myocardial ischemia/reperfusion injury (MI/RI). However, antioxidative treatment is hampered by the lack of safe and effective therapeutics. Polydopamine (PDA), as a biodegradable class of nanomaterial with excellent antioxidant properties, has shown great potential in treating MI/RI. To achieve site-specific antioxidative efficacy, we established a PDA-based biomimetic nanoplatform (PDA@M), which consisted of a polydopamine core and a macrophage membrane shell to form a shell-core structure. By inheriting the inherent migration capability of macrophages, PDA@M was able to target the infarcted myocardium and exert an antioxidative effect to protect the myocardium. The results demonstrated that the accumulation of the membrane-wrapped nanoparticles (NPs) in the infarcted myocardium was greatly increased as compared with PDA alone, which effectively relieved the MI/RI-induced oxidative stress. PDA@M largely decreased the infarct size and improved the cardiac function post-MI/RI. Our study revealed that PDA@M could inhibit cell pyroptosis by suppressing the NLRP3/caspase-1 pathway, which is known to play a significant role in the antioxidant signaling pathway. In summary, PDA@M can target the infarcted myocardium and exert antioxidative and antipyroptosis functions to protect the myocardium against MI/RI-induced oxidative stress, suggesting that it may prove to be a potential therapeutic agent for MI/RI.

Risk factors for postoperative sepsis-induced cardiomyopathy in patients undergoing general thoracic surgery: a single center experience.

BACKGROUND: The current study aimed to investigate the incidence of sepsis-induced cardiomyopathy (SICM) in patients who received general thoracic surgery, along with the risk factors and management strategies for this complication. METHODS: The clinical records of 163 patients with postoperative sepsis were retrospectively reviewed. After propensity score matching, 144 patients were divided into 2 groups by stroke volume: the SICM group (n=72) and the non-SICM group (n=72). RESULTS: The overall incidence of postoperative SICM was 53.99%. Multiple logistic regression analysis showed that stroke volume and C-reactive protein were independent predictors of mortality in patients with postoperative sepsis. Statistical analysis by t-test and χ2 test indicated that mortality (P=0.000), B-type natriuretic peptide (P=0.001), left ventricular ejection fraction (P=0.000), the mitral peak velocity of early filling/early diastolic mitral annular velocity (E/e') (P=0.049), C-reactive protein (P=0.016), procalcitonin (P=0.013), serum creatinine (P=0.016), platelets (P=0.028), and lactic acid (P=0.002) were significantly associated with the occurrence of postoperative SICM. Among these parameters, B-type natriuretic peptide was identified as the best biomarker for predicting SICM by receiver operating characteristic (ROC) curve analysis. CONCLUSIONS: It is vital to improve the diagnosis and standard management of SICM. A combined strategy comprising early detection of suspected infection, adequate use of antibiotics, close monitoring, effective drainage, and supportive care may improve the outcomes of patients with postoperative SICM.

Glycation of ß-lactoglobulin combined by sonication pretreatment reduce its allergenic potential.

ß-lactoglobulin (ß-Lg) was treated through different ultrasonic power and subsequently glycated with galactose to investigate its structural changes and immunological properties, and then evaluated by high-resolution mass spectrometry, enzyme-linked immunosorbent assay and basophil histamine release test. Ultrasonication combined with glycation (UCG) modification significantly reduced the IgE/IgG-binding capacity, and the release of ß-hexosaminidase, histamine and interleukin-6, accompanied with changes in the secondary and tertiary structures. The decrease in the allergenicity of ß-Lg depended not only on the glycation of K47, 60, 83, 91 and 135 within the linear epitopes, but also on the denaturation of conformational epitopes, which was supported by the glycation-induced alterations of the secondary and tertiary structures. This study confirmed that UCG modification is a promising method for decreasing the allergenic potential of allergic proteins, which is likely to develop a practical technology to produce hypo-allergenic milk.

Antioxidant, metabolic enzymes inhibitory ability of Torreya grandis kernels, and phytochemical profiling identified by HPLC-QTOF-MS/MS.

In this study, the antioxidant activities, α-glucosidase and tyrosinase inhibitory ability of Torreya grandis kernels (TGK) were performed. Samples were extracted with various polarity of ethanol, and the major phytochemical profile was characterized. The results showed that 70% of ethanol extract gave the richest phenolics and flavonoids. The strongest DPPH· and ABTS·+ scavenging ability, as well as the best inhibition on tyrosinase and α-glucosidase was also detected on 70% of ethanol extract. Among the fractions of 70% of ethanol extract, the ethyl acetate fraction (EAF) owned the highest phenolics, flavonoids, and the best DPPH· and ABTS·+ scavenging ability, and tyrosinase inhibition. Unexpectedly, the dichloromethane fraction possessed the strongest inhibition on α-glucosidase, which was much greater than that of acarbose. HPLC-QTOF-MS/MS analysis result to the characterization of 19 compounds from EAF. The results implied that TGK can be a potential source of natural antioxidants, α-glucosidase and tyrosinase inhibitors. Practical applications The kernels of T. grandis are one of the precious nuts in the world, and the extracts were advertised to show a variety of biological activities and pharmacological effects. However, researches on the phytochemical constituents and bioactivities are fewer. In this study, TGK was found to show good potency in antioxidant, α-glucosidase and tyrosinase inhibitory activities. The 70% ethanol is the best solvent for extracting above mentioned active components, and ethyl acetate can be the suitable enriching solvent. In addition, the predominant phytochemical compounds in EAF were characterized. Therefore, this research can help to the performance of further research and application of TGK in functional products.

The Stroke Stigma Scale: a reliable and valid stigma measure in patients with stroke.

OBJECTIVE: The objective of the study is to analyse the psychometric properties of the Stroke Stigma Scale, a novel scale to assess perceived stigma of patients with stroke. DESIGN: This is a psychometric study. SETTING: Neurology or rehabilitation units in three hospitals in China. SUBJECTS: A total of 288 patients with stroke. INTERVENTIONS: None. MEASURES: The content validity of the Stroke Stigma Scale was assessed through expert consultation. Criterion validity was evaluated based on the scale's relationships with the Stigma Scale for Chronic Illness and the Self-rating Depression Scale. Construct validity was assessed using exploratory factor analysis, and internal consistency was tested with Cronbach's α. RESULTS: The final version Stroke Stigma Scale consists of 16 items. It showed strong positive correlations with both the Stigma Scale for Chronic Illness (ρ = 0.89, P < 0.001) and the Self-rating Depression Scale (ρ = 0.82, P < 0.001). The exploratory factor analysis revealed four components of the Stroke Stigma Scale: internalized stigma, physical impairment, discrimination experience, and social isolation, which were strongly associated with our perceived stroke stigma model. Cronbach's α for the total scale was 0.92, and that of each subscale was 0.77-0.86. The test-retest reliability with intra-class correlation coefficients of the total scale was 0.92 (P < 0.001), and intra-class correlation coefficients of each subscale were 0.74-0.89 (P < 0.001). CONCLUSIONS: The Stroke Stigma Scale is a reliable and valid measure of perceived stigma in patients with stroke, which may be useful in stigma prevention and stroke rehabilitation.

Stigma experienced by Chinese patients with stroke during inpatient rehabilitation and its correlated factors: a cross-sectional study.

Background: Stroke-related stigma can have a negative effect on patients, and is associated with stereotyping, prejudice, and discrimination; however, the exact stigma experienced by patients remains ambiguous. Objectives: To evaluate the stigma experienced by patients with stroke, determine associated factors, and explore relationships between stigma and early rehabilitation. Methods: Overall, we examined 288 patients with stroke. Patient characteristics were determined through medical records and investigations, while stigma status (comprising total stigma, internalized stigma, and enacted stigma), depression level, functional independence, and functional outcome were also assessed. Demographic and clinical characteristics were evaluated through univariate analysis, and significant variables were further analyzed through linear regression. The relationships between stigma and early rehabilitation (depression, functional independence, and functional outcomes) were also analyzed. Results: The sample's total stigma, internalized stigma, and enacted stigma scores were 47.76 ± 18.00, 30.07 ± 12.25, and 17.69 ± 6.37, respectively. Employment status before stroke, caregivers, physical impairment, and number of impairments were all relevant to all types of stigma (P< .05). The regression analysis showed that caregivers and physical impairment are the two main predictors of total, internalized, and enacted stigma (P< .01). The correlation analysis revealed that stigma is associated with depression (r = 0.671 ~ 0.690, P< .001), functional independence (r = -0.562~-0.707, P< .001), and functional outcomes (r = 0.436 ~ 0.637, P< .001). Conclusions: Stigma was moderate, and internalized stigma may be more apparent; therefore, physicians should pay more attention to patients who report or show signs of experiencing stroke-related stigma.

Up-regulation of growth factor independence 1 in endotoxin tolerant macrophages with low secretion of TNF-alpha and IL-6.

OBJECTIVE: Endotoxin tolerance refers to a low response to lipopolysaccharide (LPS). We hypothesized that growth factor independence 1 (Gfi1) involves in the endotoxin tolerance in macrophages. METHODS: Endotoxin tolerance was induced in the RAW264.7 cell line and thioglycolate-elicited murine peritoneal macrophages by incubation with 100 ng/ml LPS for 20 h. Macrophages without the pretreatment were set as control. Both endotoxin tolerant and control cells were then stimulated with 1,000 ng/ml LPS for indicated period of incubation. Gfi1 mRNA expression and protein production were investigated by real-time PCR and Western blotting, respectively. ELISA was performed to quantify the secretion of TNF-alpha and IL-6. RESULT: Compared with non-endotoxin tolerant macrophages, endotoxin tolerant cells secreted a lower amount of TNF-alpha and IL-6. The mRNA expression of Gfi1 in endotoxin tolerant macrophages was higher than that of control in both RAW264.7 cells and thioglycolate-elicited murine peritoneal macrophages. The protein production was accordingly up-regulated in endotoxin tolerant RAW264.7 cells. CONCLUSION: In in vitro endotoxin tolerant macrophages, the expression of Gfi1 mRNA and protein were up-regulated after high dose LPS stimulation, accompanied with a blunted TNF-alpha and IL-6 secretion. Gfi1 might participate in the mechanism of endotoxin tolerance.