Search

This study aimed to explore the role of p53/p21 in osteoarthritis (OA). OA animal model was established by the anterior cruciate ligamentotomy (ACLT). 24 rats were randomly divided into control, OA, OA+p53 inhibitor and OA+pyroptosis inducer groups (n = 6). In the knee joint tissue, microstructural changes were analysed by Micro-CT. Histopathological changes were stained by HE and safranin-fast green. NLRP3 and Caspase-1 were detected by immunohistochemistry. The chondrocytes C-28I2 were divided into control, LPS+ ATP and p53 inhibitor groups. The cell viability, apoptosis, and LDH release were measured by MTT assay, TUNEL staining and LDH kit. The expression of p53/p21 and pyroptosis pathways were examined by western blot. The p53 inhibitor reduced the relative volume of trabecular bone (BV/TV) and trabecular bone thickness (Tb.Th), while increased trabecular separation (Tb.Sp). Moreover, the p53 inhibitor improved histopathological changes in the knee joint, attenuated cartilage damage, and reduced the expression of p53/p21 and pyroptosis pathways-related proteins. In vitro assay showed that the p53 inhibitor increased C-28I2 cell activity, reduced LDH release and apoptosis and reduced p53/p21 and pyroptosis pathways-related proteins. Totally, p53 inhibitors improved the cartilage tissue and chondrocyte damage, inhibited cell pyroptosis and the progression of OA.

Purpose This study aims to explore the innovative strategies of business model of the free-to-fee switch, the relationship between the business model innovation and customer knowledge and further develop a conceptual model. Design/methodology/approach This study adopts a multiple case study method with abductive research logic, following the replication logic to select samples. A total of eight outstanding companies with altogether 312 free-to-fee switch events were selected from 1998 to 2021. Findings A strategic matrix with four innovative business models for the free-to-fee switch is generated. The parallelism between the models and customer knowledge orientations is also found. Further, the study develops the conceptual model regarding customer knowledge orientation as a key mediation. Research limitations/implications The study highlights the conceptualization definition of customer knowledge orientation and its mediation effect to the business model innovation of free-to-fee switch, which is a new issue compared with previous research. Furthermore, it reveals that there exists organizational ambidexterity, which brings a new definition of customer knowledge orientation. Practical implications This study suggests how to integrate customer knowledge orientations to support the marketing process of the business model of free-to-fee switch. It also proposes a specific mechanism to conduct the free-to-fee switch with the introduction of four innovative strategic models and eight evolutional paths. Originality/value This study creatively proposes the strategic matrix and the conceptual model of business model innovation of free-to-fee switch. Moreover, a new conceptual definition of customer knowledge orientation is specified.

This study aimed to investigate the oncologic outcomes of video-assisted thoracoscopic (VATS) left upper trisegmentectomy (LTS) vs left upper lobectomy (LUL) for patients presenting with stage I non-small cell lung cancer (NSCLC).A retrospective analysis identified 1543 consecutive patients presenting to Shanghai Pulmonary Hospital (Shanghai, China) with NSCLC for VATS LTS or LUL from 2013 to 2017. After propensity-score matching for patient demographics and tumor characteristics, 273 pairs were identified. Disease-free survival (DFS) and overall survival (OS) were estimated by the Kaplan-Meier method and compared using the log-rank test.The median follow-up time was 51.5 months. There were no significant differences in operative duration (2.11 ± 0.64 hours vs 2.49 ± 5.96 hours; P = .30), total blood loss (106.19 ± 170.83 mL vs 97.07 ± 149.34 mL; P = .51), and operative complications (10% vs 8%; P = .37) between the LUL and LTS groups. Patients undergoing LUL had longer postoperative hospital stays (5.55 ± 3.00 days vs 4.87 ± 2.33 days; P = .003), greater tumor margin distance (3.3 ± 1.2 cm vs 3.1 ± 0.9 cm; P.001), and greater number of lymph nodes harvested (8.0 ± 3.2 vs 6.8 ± 3.3; P.001) than patients undergoing LTS, but the margin-to-tumor ratio was not statistically different (2.5 ± 1.6 vs 2.3 ± 1.1; P = 0.11). Median DFS (49.5 months vs 54.3 months; P = .77) and OS (49.5 months vs 55.0 months; P = .88) were not significantly different between patients undergoing LTS and those undergoing LUL, and similar outcomes were noted across subgroups of patients stratified by tumor stage, pathologic type, and radiographic manifestations.VATS LTS and LUL had comparable oncologic outcomes for stage I NSCLC, regardless of tumor pathologic types and radiologic findings, as long as negative margins were confirmed.

Long-segment tracheal resection is technically challenging due to its high tension during reconstruction. Therefore, tracheal release maneuvers, including pulmonary hilar release and pericardial dissection, were required to reduce the anastomotic tension. Traditional hilar release is performed via thoracotomy; however, this approach is invasive. We report a case of bilateral hilar and pericardial release via a video-assisted thoracoscopic surgery (VATS) approach in resecting a long segment tracheal adenoid cystic carcinoma that spanned 50% of the trachea. The bilateral hilar and pericardial releases were performed under general anesthesia through biportal VATS, which contributed to a total of 2.5 cm of additional mobility to the trachea, and the infrahyoid release was then performed through a cervical collar incision. The tumor-involved tracheal segments were removed via median sternotomy, totaling 6.0 cm in length, and the remaining trachea could be successfully reconstructed with a tension-free anastomosis. The total operative duration was 4.5 hours. The patient suffered a transient swallowing dysfunction during the postoperative course, with a good luminal patency in the trachea after 1 month postoperatively. Therefore, bilateral hilar release via VATS can be considered to be a less invasive, avoiding the potential complications related to a thoracotomy, but similarly effective release maneuver for long-segment tracheal resections.

Formation of epitaxial heterostructures via post-growth self-assembly is important in the design and preparation of functional hybrid systems combining unique properties of the constituents. This is particularly attractive for the construction of metal halide perovskite heterostructures, since their conventional solution synthesis usually leads to non-uniformity in composition, crystal phase and dimensionality. Herein, we demonstrate that a series of two-dimensional and three-dimensional perovskites of different composition and crystal phase can form epitaxial heterostructures through a ligand-assisted welding process at room temperature. Using the CsPbBr

Pulmonary aspergilloma is chronic and invasive, potentially leading to life-threatening massive hemoptysis. The role of surgery for treating pulmonary aspergilloma and its effect on long-term survival needs more study.We reviewed 166 patients with aspergillomas treated at Shanghai Pulmonary Hospital from 2004 to 2017. Surgery indications included destroyed lung parenchyma, recurrent hemoptysis, despite appropriate medical treatment, and isolated pulmonary nodules suspected to be aspergilloma. Pulmonary aspergillomas are classified as simple (in an isolated thin-walled cavity, ≤3 mm) or complex (a thick-walled cyst,3 mm) based on computed tomographic scan findings.Aspergilloma was complex in 100 patients (60.2%) and simple in 66 (39.8%). The median (interquartile range) size of complex aspergillomas (2.5 [0.3-8.0] cm) was larger than that (2.0 [0.2-6.0] cm) of simple types (P.001). Hemoptysis occurred in 72 patients (72%) with complex disease and in 35 (53%) with simple disease (P = .014). Video-assisted thoracoscopic surgery was performed in 42 patients (63.6%) with simple aspergillomas, and 75 patients (75%) with of complex aspergillomas underwent thoracotomy. Prolonged air leakage (7 days) was the most common (17 [10.2%]) postoperative complication. A bronchopleural fistula developed postoperatively in 1 patient (0.6%). One patient (0.6%) died of respiratory failure within 30 days postoperatively. Two patients (1.2%) experienced recurrence during follow-up. The overall 10-year survival rates were 87.7% for complex aspergillomas and 94.97% for simple aspergillomas (P = .478). Diabetes (12 [7.2%]; hazard ratio, 13.15; 95% CI, 1.12-154.46) was associated with a worse prognosis.The perioperative morbidity and mortality of pulmonary aspergillomas are acceptable. Overall survival rates of simple and complex types are comparable.

Acute kidney injury (AKI) results in high morbidity and mortality among inpatients, while effective treatment and intervention are still absent. Therefore, this study aims to explore the effects of Scutellarin (Scu) in experimental models in vivo and in vitro.In vivo experiment, we employed a total of 30 Wistar rats, which further were modelled by a bilateral renal pedicle clip for 45 min, then received 50 mg/kg/day Scu. In vitro, HK-2 cells were administered with 20μΜ Scu and then incubated in hypoxia/reoxygenation (H/R) conditions for 24 h. The knockdown of Nrf2 expression was conducted by small interfering RNA (siRNA) transfection.As a result, the AKI model was well established with an increased SCr, BUN, KIM-1 level, and histological injury score, while Scu treatment reduced the levels above and increased the antioxidative enzyme HO-1. H/R induced an increase of ROS in HK-2 cells, while Scu decreased the ROS level. Bioinformatics results showed the transcription factor Nrf2 was a hub protein during the AKI, which also bound to Scu with low binding energy, indicating that the downstream effect of Scu might be mediated by Nrf2. To verify the suppose above, we employed siRNA against Nrf2, which shows a significant increase in ROS after Nrf2 was blocked. Meanwhile, the HO-1 showed similar expression compared with the 'H/R + Nrf2 siRNA' and 'H/R + Nrf2 siRNA + Scu' group, implying the protective effect of Scu was mediated by the Nrf2/HO-1 pathway.Scu led to up-regulation of HO-1 through activating the Nrf2 signalling pathway, protecting the kidneys from ischemia/reperfusion (I/R)-induced oxidative damage.

An organocatalyzed, formal (3+3) cycloaddition reaction is described for the practical synthesis of substituted pyridines. Starting from readily available enamines and enal/ynal/enone substrates, the protocol affords tri- or tetrasubstituted pyridine scaffolds bearing various functional groups. This method was demonstrated on a 50 g scale, enabling the synthesis of 2-isopropyl-4-methylpyridin-3-amine, a raw material used for the manufacture of sotorasib. Mechanistic analysis using two-dimensional nuclear magnetic resonance (NMR) spectrometry revealed the transformation proceeds through the reversible formation of a stable reaction off-cycle species that precedes pyridine formation.

RNA-directed DNA methylation (RdDM) and ROS1-dependent active DNA demethylation pathways are antagonistic processes that dynamically regulate site-specific methylation. In this study, we obtained a mutant with reduced luciferase (LUC) luminescence by genetic screening, which was named rll5-1 (for reduced LUC luminescence 5-1). The rll5-1 mutant showed narrower, frizzled and curly leaves, and the low-LUC-luminescence phenotype in the rll5-1 mutant can be largely restored by DNA methylation inhibitor 5-Aza-2'-deoxycytidine. Map-based cloning coupled with genome resequencing data revealed that a nucleotide substitution of G to A was found at the 124th bp of ORF of At4G10190, leading to an aspartate-to-asparagine change at position 42 in such a protein. Bisulfite sequencing data indicated that DNA methylation of 3' region of the double 35S promoter that drives the LUC expression was appreciably increased. Further analysis revealed that there were 4747 hypo-DMRs and 936 hyper-DMRs found in the rll5-1 genome, and the hypo-DMRs was predominantly distributed on TEs, which appeared to stem from the downregulation of a few RdDM pathway genes and DNA methyltransferase genes. Closer inspection demonstrated that there were 1229 hypo-DMRs commonly shared among rll5-1, nrpd1-3 and nrpe1-11, and a total of 1349 hypo-DMRs were common to rll5-1 and cmt2 mutants. Thus, these studies demonstrate the roles of RLL5 in preventing transgene silencing and in maintaining genome-wide DNA methylation in a direct/indirect or locus-specific manner.

The negative effects of low temperature can readily induce a variety of diseases. We sought to understand the reasons why cold stress induces disease by studying the mechanisms of fine-tuning in macrophages following cold exposure. We found that cold stress triggers increased macrophage activation accompanied by metabolic reprogramming of aerobic glycolysis. The discovery, by genome-wide RNA sequencing, of defective mitochondria in mice macrophages following cold exposure indicated that mitochondrial defects may contribute to this process. In addition, changes in metabolism drive the differentiation of macrophages by affecting histone modifications. Finally, we showed that histone acetylation and lactylation are modulators of macrophage differentiation following cold exposure. Collectively, metabolism-related epigenetic modifications are essential for the differentiation of macrophages in cold-stressed mice, and the regulation of metabolism may be crucial for alleviating the harm induced by cold stress.

Supplementary Figures and Figure Legends

Supplementary Table 1 Primer sequences

Induction of ferroptosis, a recently defined form of nonapoptotic cell death caused by iron-dependent lipid peroxidation, has emerged as an anticancer strategy. Erastin is a ferroptosis activator that promotes cell death that not only depends on the depletion of cellular cysteine but also relies on mitochondrial oxidative metabolism of glutamine. Here, we demonstrate that ASS1, a key enzyme involved in the urea cycle, plays a crucial role in ferroptosis resistance. Loss of ASS1 increased the sensitivity of non–small cell lung cancer (NSCLC) cells to erastin in vitro and decreased tumor growth in vivo. Metabolomics analysis with stable isotope-labeled glutamine showed that ASS1 promotes reductive carboxylation of cytosolic glutamine and compromises the oxidative tricarboxylic acid cycle from glutamine anaplerosis, reducing mitochondrial-derived lipid reactive oxygen species. Moreover, transcriptome sequencing showed that ASS1 activates the mTORC1–SREBP1–SCD5 axis to promote de novo monounsaturated fatty acid synthesis by using acetyl-CoA derived from the glutamine reductive pathway. Treating ASS1-deficient NSCLC cells with erastin combined with arginine deprivation significantly enhanced cell death compared with either treatment alone. Collectively, these results reveal a previously unknown regulatory role of ASS1 in ferroptosis resistance and provide a potential therapeutic target for ASS1-deficient NSCLC.Significance:ASS1 promotes reductive carboxylation of glutamine and confers ferroptosis resistance, providing multiple treatment options for ASS1-deficient non–small cell lung cancer.

BACKGROUND The widespread use of electronic health records in clinical and biomedical fields makes the removal of protected health information (PHI) essential to maintain privacy. However, a significant portion of information is recorded in unstructured textual form posing a challenge to de-identify. In countries like Taiwan, medical records could be written in a mixture of more than one language, referred to as code-mixing (CM). Most current clinical natural language processing techniques are designed for monolingual texts, and there is a need to address the de-identification of CM texts. OBJECTIVE The aim of this study was to investigate the effectiveness and underlying mechanism of fine-tuned PLMs in identifying PHIs in CM context. Additionally, we also aimed to evaluate the potential of prompting LLMs in recognizing PHIs in a zero-shot manner. METHODS We compiled the first clinical CM deidentification dataset consisting of texts written in Chinese and English. We explored the effectiveness of fine-tuning pre-trained language models (PLMs) in recognizing PHIs in CM content, focusing on whether PLMs exploit naming regularity and mention coverage to achieve superior performance by probing the developed models’ outputs to examine their decision-making process. Furthermore, we investigated the potential of prompt-based in-context learning of large language models (LLMs) in recognizing PHIs in CM text. RESULTS The developed methods were evaluated on a CM de-identification corpus of 1,700 discharge summaries. We observed that different PHI types had their preference in their occurrence within the different types of language-mixed sentences, and PLMs could effectively recognize PHIs by exploiting the learned name regularity. However, the models may exhibit suboptimal results when regularity was weak or mentions contain unknown words that the representations cannot generate well. We also found that the availability of CM training instances is essential for the model’s performance. Furthermore, LLM-based de-identification method is a feasible and appealing approach that can be controlled and enhanced through natural language prompts. CONCLUSIONS The study contributes to understanding the underlying mechanism of PLMs in addressing the de-identification process in CM context and highlights the significance of incorporating CM training instances into the model training phase. The LLM-based de-identification method is a feasible approach, but carefully crafted prompts are essential to avoid unwanted output. However, the use of such methods in the hospital setting requires careful consideration of data security and privacy concerns. Further research could explore the augmentation of PLMs and LLMs with external knowledge to improve their strength in recognizing rare PHIs.

Diethyldithiocarbamate (DDTC) has been employed in the sulfide ore flotation process, due to its excellent collection performance. Herein, we investigated the interfacial adsorption behavior of DDTC on four mineral phases of high-sulfur residue — sulfur, pyrite, sphalerite and lead sulfate. The adsorption behavior — adsorption structure and energy and electron localization function cross section — of DDTC and H2O were explored using density function theory calculation. The result was helpful to construct a co-adsorption model of DDTC and H2O, which was validated by the pure minerals flotation operation and the characterization of fourier transform infrared spectra. The co-adsorption model indicated that the DDTC adsorption on the sulfur, sphalerite and lead sulfate was weak with physical bonding, while its adsorption on the pyrite was strong with chemical bonding. Practical bench-scale operation of high-sulfur residue flotation was performed, whose result was different from that obtained from the pure mineral flotation. To explain the difference, our developed model predictions and the mineral fugacity pattern analysis on the high-sulfur residue flotation were synergistically used. Overall, this work first proposed the co-adsorption model of DDTC and H2O and provided important insights into the interfacial adsorption in the high-sulfur residue flotation.

Supplementary Figure S2 shows binding of REGN3767 to T cells

In the tumor microenvironment, multiple inhibitory checkpoint receptors can suppress T-cell function, thereby enabling tumor immune evasion. Blockade of one of these checkpoint receptors, PD-1, with therapeutic antibodies has produced positive clinical responses in various cancers; however, the efficacy of this approach can be further improved. Simultaneously targeting multiple inhibitory checkpoint receptors has emerged as a promising therapeutic strategy. Here, we report the development and characterization of REGN3767, a fully human IgG4 antibody targeting LAG-3, another inhibitory receptor on T cells. REGN3767 binds human and monkey LAG-3 with high affinity and specificity and blocks the interaction of LAG-3 with its ligand, MHC class II. In an engineered T-cell/antigen-presenting cell bioassay, REGN3767 alone, or in combination with cemiplimab (REGN2810, human anti-PD-1 antibody), blocked inhibitory signaling to T cells mediated by hLAG-3/MHCII in the presence of PD-1/PD-L1. To test the in vivo activity of REGN3767 alone or in combination with cemiplimab, we generated human PD-1xLAG-3 knockin mice, in which the extracellular domains of mouse Pdcd1 and Lag3 were replaced with their human counterparts. In these humanized mice, treatment with cemiplimab and REGN3767 showed increased efficacy in a mouse tumor model and enhanced the secretion of proinflammatory cytokines by tumor-specific T cells. The favorable pharmacokinetics and toxicology of REGN3767 in nonhuman primates, together with enhancement of antitumor efficacy of anti-PD-1 antibody in preclinical tumor models, support its clinical development.

Supplementary Table S2 shows PK data for REGN3767 in monkeys

Tab S1-S18

Purpose: Effective therapies for the majority of metastatic acral melanoma patients have not been established. Thus, we investigated genetic aberrations of CDK4 pathway in acral melanoma and evaluated the efficacy of CDK4/6 inhibitors in targeted therapy of acral melanoma.Experimental Design: A total of 514 primary acral melanoma samples were examined for the copy number variations (CNV) of CDK4 pathway-related genes, including Cdk4, Ccnd1, and P16INK4a, by QuantiGenePlex DNA Assay. The sensitivity of established acral melanoma cell lines and patient-derived xenograft (PDX) containing typical CDK4 aberrations to CDK4/6 inhibitors was evaluated.Results: Among the 514 samples, 203 cases, 137 cases, and 310 cases, respectively, showed Cdk4 gain (39.5%), Ccnd1 gain (26.7%), and P16INK4a loss (60.3%). The overall frequency of acral melanomas that contain at least one aberration in Cdk4, Ccnd1, and P16INK4a was 82.7%. The median overall survival time for acral melanoma patients with concurrent Cdk4 gain with P16INK4a loss was significantly shorter than that for patients without such aberrations (P = 0.005). The pan-CDK inhibitor AT7519 and selective CDK4/6 inhibitor PD0332991 could inhibit the cell viability of acral melanoma cells and the tumor growth of PDX with Cdk4 gain plus Ccnd1 gain, Cdk4 gain plus P16INK4a loss, and Ccnd1 gain plus P16INK4a loss.Conclusions: Genetic aberration of CDK4 pathway is a frequent event in acral melanoma. Acral melanoma cell lines and PDX containing CDK4 pathway aberrations are sensitive to CDK4/6 inhibitors. Our study provides evidence for the testing of CDK4/6 inhibitors in acral melanoma patients. Clin Cancer Res; 23(22); 6946–57. ©2017 AACR.

Fig S1-S5

PDF file - 2243K, Figure S3. Genetic mutations in exon 18 of PDGFRA.

Supplementary Figure1-10, Table1-2 Figure S1. The relations of REGγ with Hippo-YAP signal pathway in colon cancer. A. The protein levels of Lats2 and p-Lats1 were unchanged in HCT116 and HT29 human colon cancer cells with REGγ knockdown. The expression of REGγ, Lats2 and p-Lats1 were measured by Western Blot. B. REGγ knockdown did not change the levels of p-YAP (S397) in HCT116 and HT29 human colon cancer cells. Figure S2. REGγ promotes degradation of Lats1. A. Silencing REGγ slowed down degradation of endogenous Lats1. REGγ sh-N or sh-R HCT116 cells were treated with cycloheximide (100μg/ml) for indicated time followed by Western Blotting. Quantitated results were plotted to indicate dynamic changes (Analysis of Variance, n=3, *p0.05) Figure S9. A Model of crosstalk among REGγ, Hippo-Yap, and NF-κB. In human colon cancer cells, overexpression of REGγ promotes the degradation of Lats1, thus activating YAP signaling. YAP activation empowers a reciprocal positive regulation with NF-κB, leading to aberrant cell proliferation and development of inflammationassociated colon cancer. In contrast, cells with REGγ depletion are resistant to Lats1 inactivation, inhibiting YAP activation and its crosstalk to NF-κB to prevent tumor formation Figure S10. P53 was not affected by REGγ deficiency in human colon cancer cells. A. Western Blot analysis for REGγ and p53 in HCT116 sh-N and sh-R human colon cancer cells. β-actin served as a loading control. B. Western Blot analysis of p53 from mouse normal colon and tumor tissues of REGγ+/+ and REGγ-/- mice showed no marked changes. All experiments were repeated three times. Table 1. Sequences of primers used for Q-PCR Table 2. Sequences of primers used for ChIP

Supplementary Data Supplementary Table S1.Clinical pathological parameters of enrolled patients Supplementary Table S2. The sample information of QuantiGenePlex RNA assay Supplementary Table S3. Correlation of CDK4 pathway aberrations withPD-L1 protein expression in melanoma patients.

Supplementary Table S1. Genetic aberrations of cells used in this study Supplementary Table S2. Clinicopathological features and genetic aberrations of established PDX models used in this study Supplementary Table S3. Detailed clinicopathological features and genetic aberrations of patients Supplementary Table S4. Validation of copy number variations of genes related to CDK4 pathway by Q-PCR and mutation status of therapeutic targets Supplementary Table S5. Correlation between copy number variations and protein expression of CDK4 pathway in acral melanoma Supplementary Table S6. Correlation of concurrent CDK4 pathway aberrations to clinicopathologic features of acral melanoma Supplementary Table S7. Univariate and multivariate analysis of risk factors associated with overall survival Supplementary Table S8. Correlation of concurrent CDK4 pathway aberrations to mitotic rate and TILs score of acral melanoma

PDF file - 121K, Supplementary Table S1. Primers and annealing temperatures for the sequencing of PDGFRA.