Youths' awareness and attitudes towards raising the minimum legal age of smoking and passive smoking in Singapore.

Objectives: Early smoking initiation has been associated with a higher risk of developing long-term smoking habit. There is a growing global consensus that demands raising the minimum legal age (MLA) for smoking as an approach to address this problem. Singapore successfully raised the MLA from 18 to 21 years in 2021. This study aimed to evaluate the awareness and attitude of multi-ethnic Asian youth (aged 15-24) on raising MLA to 21 and passive smoking. Methods: A cross-sectional survey comprising of 23 items was circulated via a secure internet-based platform, FORMSG between September and November 2022. Data were analyzed for descriptive statistics. Categorical variables were compared for association with receptivity toward change in MLA using Chi-Squared test and multivariable logistic regression analysis using Rstudio. Post-hoc Bonferroni correction were further utilized for pairwise comparison. Results: Majority (80.3%) of the 608 participants expressed their support for MLA 21 implementation. Participants' age was a significant variable as those aged 15-17 years old (OR = 2.1, 95%CI = 1.01-4.32, p = 0.048) showed a higher likelihood of supporting MLA implementation compared to those aged 21 and above. In addition, majority (89.8%) of them were also aware of the harmful effects of passive smoking. When it came to discouraging smoking among youth, family influence (64%) and school education (55.6%) emerged as the top strategies. Conclusion: Most of the youth express strong support for raising the MLA to 21, with over 80% in favor of such change, reflects a significant harmony among youth in favor of tobacco-free environment.

Proteomic Identification of Small Extracellular Vesicle Proteins LAMB1 and Histone H4 for Prostate Cancer Diagnosis and Risk Stratification.

Diagnosis and stratification of prostate cancer (PCa) patients using the prostate-specific antigen (PSA) test is challenging. Extracellular vesicles (EVs), as a new star of liquid biopsy, has attracted interest to complement inaccurate PSA screening and invasiveness of tissue biopsy. In this study, a panel of potential small EV (sEV) protein biomarkers is identified from PCa cell lines using label-free LC-MS/MS proteomics. These biomarkers underwent further validation with plasma and urine samples from different PCa stages through parallel reaction monitoring-based targeted proteomics, western blotting, and ELISA. Additionally, a tissue microarray containing cancerous and noncancerous tissues is screened to provide additional evidence of selected sEV proteins associated with cancer origin. Results indicate that sEV protein LAMB1 is highly expressed in human plasma of metastatic PCa patients compared with localised PCa patients and control subjects, while sEV protein Histone H4 is highly expressed in human urine of high-risk PCa patients compared to low-risk PCa patients and control subjects. These two sEV proteins demonstrate higher specificity and sensitivity than the PSA test and show promise for metastatic PCa diagnosis, progression monitoring, and risk stratification.

Corneal Sub-Basal Nerve Plexus Regeneration Pattern following Implantable Collamer Lens Implantation for Myopia: A Prospective Longitudinal In Vivo Confocal Microscopy Study.

Implantable Collamer Lens (ICL) surgery has increasingly been adopted for myopia correction in recent decades. This study, employing in vivo confocal microscopy (IVCM), aimed to assess the impact of corneal incision during ICL surgery on the corneal sub-basal nerve plexus (SNP) and adjacent immune dendritiform cells (DCs). In this longitudinal study, eyes from 53 patients undergoing ICL surgery were assessed preoperatively and postoperatively over a twelve-month period. Quantification of seven SNP parameters was performed using ACCMetrics V.2 software. Ultimately, the final analysis was restricted to one eye from each of the 37 patients who completed a minimum of three months' postoperative follow-up. Preoperative investigations revealed a positive correlation of DC density with patient age and a negative association with corneal nerve fiber density (CNFD). Additionally, both DCs and CNFD were positively linked to spherical equivalent refraction (SER) and inversely related to axial length (AL). Intriguingly, preoperative DC density demonstrated an indirect relationship with both baseline and postoperative CNFD changes. Post-surgery, an initial surge in DC density was observed, which normalized subsequently. Meanwhile, parameters like CNFD, corneal nerve fiber length (CNFL), and corneal nerve fractal dimension (CNFrD) initially showed a decline following surgery. However, at one-year follow-up, CNFL and CNFrD displayed significant recovery, while CNFD did not return to its baseline level. This study thus delineates the regeneration pattern of SNP and alterations in DC density post-ICL surgery, highlighting that CNFD in the central cornea does not completely revert to preoperative levels within a year. Given these findings, practitioners are advised to exercise caution in older patients, those with high myopia, or elevated preoperative DCs who may undergo delayed SNP regeneration.

Unlocking the Potential of Pterostilbene: A Pharmaceutical Element for Aptamer-Based Nanomedicine.

Natural compounds like pterostilbene (PTE) have gained recognition for their various biological activities and potential health benefits. However, challenges related to bioavailability and limited clinical efficacy have prompted efforts to strengthen their therapeutic potential. To meet these challenges, we herein rationally designed and successfully synthesized a pharmaceutical phosphoramidite that allows for the programmable incorporation of PTE into oligonucleotides. The resultant aptamer-PTE conjugate can selectively bind to cancer cells, leading to a specific internalization and drug release. Moreover, compared with free PTE, the conjugate exhibits superior cytotoxicity in cancer cells. Specifically, in a zebrafish xenograft model, the nanomedicine effectively inhibits tumor growth and neovascularization, highlighting its potential for targeted antitumor therapy. This approach presents a promising avenue for harnessing the therapeutic potential of natural compounds via a nanomedicine solution.

Effect of different iron ratios on interaction and thermodynamic stability of bound whey protein isolate.

Whey protein isolates (WPI) are known to have mineral-binding capacity to promote iron absorption. The aim of this study was to investigate the effect of iron ratio on the conformational structure of iron-bound whey protein isolate (WPI-Fe) and its thermodynamic stability. It was shown that the iron to protein ratio affects both the iron binding capacity of WPI and the iron valence state on the surface of WPI-Fe complexes. As the iron content increases, aggregation between protein molecules occurs. In addition, WPI-Fe nanoparticles have thermodynamic stability and Fe2+ has a high affinity with WPI for spontaneous exothermic reactions. This study demonstrates that WPI-Fe complexes can be used to efficiently deliver high-quality iron source (Fe2+) for future iron supplements.

Proteomic-based stratification of intermediate-risk prostate cancer patients.

Gleason grading is an important prognostic indicator for prostate adenocarcinoma and is crucial for patient treatment decisions. However, intermediate-risk patients diagnosed in the Gleason grade group (GG) 2 and GG3 can harbour either aggressive or non-aggressive disease, resulting in under- or overtreatment of a significant number of patients. Here, we performed proteomic, differential expression, machine learning, and survival analyses for 1,348 matched tumour and benign sample runs from 278 patients. Three proteins (F5, TMEM126B, and EARS2) were identified as candidate biomarkers in patients with biochemical recurrence. Multivariate Cox regression yielded 18 proteins, from which a risk score was constructed to dichotomize prostate cancer patients into low- and high-risk groups. This 18-protein signature is prognostic for the risk of biochemical recurrence and completely independent of the intermediate GG. Our results suggest that markers generated by computational proteomic profiling have the potential for clinical applications including integration into prostate cancer management.

A pathway activity-based proteomic classifier stratifies prostate tumors into two subtypes.

Prostate cancer (PCa) is the second most common cancer in males worldwide. The risk stratification of PCa is mainly based on morphological examination. Here we analyzed the proteome of 667 tumor samples from 487 Chinese PCa patients and characterized 9576 protein groups by PulseDIA mass spectrometry. Then we developed a pathway activity-based classifier concerning 13 proteins from seven pathways, and dichotomized the PCa patients into two subtypes, namely PPS1 and PPS2. PPS1 is featured with enhanced innate immunity, while PPS2 with suppressed innate immunity. This classifier exhibited a correlation with PCa progression in our cohort and was further validated by two published transcriptome datasets. Notably, PPS2 was significantly correlated with poor biochemical recurrence (BCR)/metastasis-free survival (log-rank P-value < 0.05). The PPS2 was also featured with cell proliferation activation. Together, our study presents a novel pathway activity-based stratification scheme for PCa.

Fabrication of cobalt-zinc bimetallic oxides@polypyrrole composites for high-performance electromagnetic wave absorption.

Composite materials that combine magnetic and dielectric losses offer a potential solution to enhance impedance match and significantly improve microwave absorption. In this study, Co3O4/ZnCo2O4 and ZnCo2O4/ZnO with varying metal oxide compositions are successfully synthesized, which are achieved by modifying the ratios of Co2+ and Zn2+ ions in the CoZn bimetallic metal-organic framework (MOF) precursor, followed by a high-temperature oxidative calcination process. Subsequently, a layer of polypyrrole (PPy) is coated onto the composite surfaces, resulting in the formation of core-shell structures known as Co3O4/ZnCo2O4@PPy (CZCP) and ZnCo2O4/ZnO@PPy (ZCZP) composites. The proposed method allows for rapid adjustments to the metal oxide composition within the inner shell, enabling the creation of composites with varying degrees of magnetic losses. The inclusion of PPy in the outer shell serves to enhance the bonding strength of the entire composite structure while contributing to conductive and dielectric losses. In specific experimental conditions, when the loading is set at 50 wt%, the CZCP composite exhibits an effective absorption bandwidth (EAB) of 5.58 GHz (12.42 GHz-18 GHz) at a thickness of 1.53 mm. Meanwhile, the ZCZP composite demonstrates an impressive minimum reflection loss (RLmin) of -71.2 dB at 13.04 GHz, with a thickness of 1.84 mm. This study offers a synthesis strategy for designing absorbent composites that possess light weight and excellent absorptive properties, thereby contributing to the advancement of electromagnetic wave absorbing materials.

Fabrication of hierarchical reduced graphene oxide decorated with core-shell Fe3O4@polypyrrole heterostructures for excellent electromagnetic wave absorption.

The design of heterostructures with reasonable chemical composition and spatial structure is one of the effective strategies to achieve high performances electromagnetic wave (EMW) absorption. Herein, reduced graphene oxide (rGO) nanosheets decorated with hollow core-shell Fe3O4@PPy (FP) microspheres have been prepared by the combination of hydrothermal method, in situ polymerization method, directional freeze-drying and hydrazine vapor reduction. FP acting as traps can consume EMW trapped into their interior through the magnetic and dielectric losses. RGO nanosheets forming the conductive network are served as multi-reflected layers. Moreover, the impedance matching is optimized by the synergistic effect between FP and rGO. As expected, the synthetic Fe3O4@PPy/rGO (FPG) composite shows excellent EMW absorption performances with the minimum reflect loss (RLmin) of -61.20 dB at 1.89 mm and the effective absorption bandwidth (EAB) of 5.26 GHz at 1.71 mm. The excellent performances for the heterostructure are attributed to the synergistic effect of conductive loss, dielectric loss, magnetic loss, multiple reflection loss, and optimized impedance matching. This work provides a simple and effective strategy for the fabrication of lightweight, thin and high-performances EMW absorbing materials.

The inhibitory effect and mechanism of Resina Draconis on the proliferation of MCF-7 breast cancer cells: a network pharmacology-based analysis.

Resina Draconis (RD) is known as the "holy medicine for promoting blood circulation" and possesses antitumor properties against various types of cancer, including breast cancer (BC); however, the underlying mechanism is not well understood. To explore the potential mechanism of RD against BC using network pharmacology and experimental validation, data on bioactive compounds, potential targets of RD, and related genes of BC were obtained from multiple public databases. Gene Ontology (GO) and KEGG pathway analyses were performed via the DAVID database. Protein interactions were downloaded from the STRING database. The mRNA and protein expression levels and survival analysis of the hub targets were analyzed using the UALCAN, HPA, Kaplan‒Meier mapper, and cBioPortal databases. Subsequently, molecular docking was used to verify the selected key ingredients and hub targets. Finally, the predicted results of network pharmacology methods were verified by cell experiments. In total, 160 active ingredients were obtained, and 148 RD target genes for the treatment of BC were identified. KEGG pathway analysis indicated that RD exerted its therapeutic effects on BC by regulating multiple pathways. Of these, the PI3K-AKT pathway was indicated to play an important role. In addition, RD treatment of BC seemed to involve the regulation of hub targets that were identified based on PPI interaction network analysis. Validation in different databases showed that AKT1, ESR1, HSP90AA1, CASP3, SRC and MDM2 may be involved in the carcinogenesis and progression of BC and that ESR1, IGF1 and HSP90AA1 were correlated with worse overall survival (OS) in BC patients. Molecular docking results showed that 103 active compounds have good binding activity with the hub targets, among which flavonoid compounds were the most important active components. Therefore, the sanguis draconis flavones (SDF) were selected for subsequent cell experiments. The experimental results showed that SDF significantly inhibited the cell cycle and cell proliferation of MCF-7 cells through the PI3K/AKT pathway and induced MCF-7 cell apoptosis. This study has preliminarily reported on the active ingredients, potential targets, and molecular mechanism of RD against BC, and RD was shown to exert its therapeutic effects on BC by regulating the PI3K/AKT pathway and related gene targets. Importantly, our work could provide a theoretical basis for further study of the complex anti-BC mechanism of RD.

Efficacy and safety of EGFR­TKIs plus Shenqi Fuzheng injection for non-small cell lung cancer patients with EGFR-sensitive mutations.

PURPOSE: The aim of this retrospective study is to evaluate the impact on efficacy and safety between epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) alone and in combination with Shenqi Fuzheng injection (SFI) in patients with advanced NSCLC harboring epidermal growth factor receptor (EGFR) activating mutations. METHODS: Retrospectively, information of 88 patients receiving EGFR-TKIs as first-line targeted treatment or in combination with SFI in the Affiliated Drum Tower Hospital of Nanjing University Medical College and the Affiliated Cancer Hospital of Anhui University of Science and Technology was collected. The primary endpoint was to assess progression-free survival (PFS) and safety of EGFR-TKIs alone or in combination with SFI. RESULTS: Between January 2016 and December 2019, a total of 88 patients were enrolled in this research, including 50 cases in the EGFR-TKIs single agent therapy group and 38 cases in the SFI combined with EGFR-TKIs targeted-therapy group. The median PFS (mPFS) of monotherapy group was 10.50 months (95%CI 9.81-11.19), and 14.30 months (95%CI 10.22-18.38) in the combination therapy group. Compared to the single EGFR-TKIs administration, combinational regimen with SFI exhibited a lower incidence of rash and diarrhea in patients and was even better tolerated. CONCLUSIONS: SFI combined with the first-generation EGFR-TKIs are more efficient, can prominently prolong the PFS and attenuate the adverse reactions in patients with advanced NSCLC with EGFR-sensitive mutations.

Effect of incision location on with-the-rule astigmatism correction after SMILE.

PURPOSE: To explore the influence of two different incision positions of small incision lenticule extraction (SMILE) on astigmatism correction. METHODS: A total of 86 eyes of 86 patients who underwent SMILE surgery were included in this retrospective study. The criteria included that the preoperative astigmatism axis to be within 165° to 180° or 5° to 15°. All eyes were divided into two groups according to the incision position. The incisions of 34 eyes were either located on the superior temporal side of the right eye or superior nasal side of the left eye (120° group), and 52 eyes were located on the superior side (90° group). Vector analysis was used to analyze pre- and postoperative 3-month astigmatism, while subgroup analysis was applied according to the preoperative astigmatism. RESULTS: No significant difference of preoperative parameters was found between the two groups (P > 0.05). The efficacy and safety indexes were 1.11 ± 0.16 and 1.15 ± 0.16 in the 120° group, the numbers were 1.15 ± 0.17 and 1.16 ± 0.14 in the 90° group (P = 0.629 and P = 0.871). There was no significant difference in target-induced astigmatism (TIA), TIA axis, surgically induced astigmatism (SIA) magnitude, SIA axis, distance vision (DV) magnitude, DV axis, correction index (CI), angle of error (AofE), |AofE|, magnitude of error (MofE), index of success (IOS), or flattening index (FI) between 120° group and 90° group (P > 0.05). No significant difference was found in the subgroup analysis (P > 0.05). CONCLUSIONS: The incision position has limited astigmatic effect, and the operation-induced astigmatism of SMILE surgery after 3 months was little.

Cell senescence-associated genes predict the malignant characteristics of glioblastoma.

BACKGROUND: Glioblastoma (GBM) is the most malignant, aggressive and recurrent primary brain tumor. Cell senescence can cause irreversible cessation of cell division in normally proliferating cells. According to studies, senescence is a primary anti-tumor mechanism that may be seen in a variety of tumor types. It halts the growth and spread of tumors. Tumor suppressive functions held by cellular senescence provide new directions and pathways to promote cancer therapy. METHODS: We comprehensively analyzed the cell senescence-associated genes expression patterns. The potential molecular subtypes were acquired based on unsupervised cluster analysis. The tumor immune microenvironment (TME) variations, immune cell infiltration, and stemness index between 3 subtypes were analyzed. To identify genes linked with GBM prognosis and build a risk score model, we used weighted gene co-expression network analysis (WGCNA), univariate Cox regression, Least absolute shrinkage and selection operator regression (LASSO), and multivariate Cox regression analysis. And the correlation between risk scores and clinical traits, TME, GBM subtypes, as well as immunotherapy responses were estimated. Immunohistochemistry (IHC) and cellular experiments were performed to evaluate the expression and function of representative genes. Then the 2 risk scoring models were constructed based on the same method of calculation whose samples were acquired from the CGGA dataset and TCGA datasets to verify the rationality and the reliability of the risk scoring model. Finally, we conducted a pan-cancer analysis of the risk score, assessed drug sensitivity based on risk scores, and analyzed the pathways of sensitive drug action. RESULTS: The 3 potential molecular subtypes were acquired based on cell senescence-associated genes expression. The Log-rank test showed the difference in GBM patient survival between 3 potential molecular subtypes (P = 0.0027). Then, 11 cell senescence-associated genes were obtained to construct a risk-scoring model, which was systematically randomized to distinguish the train set (n = 293) and the test set (n = 292). The Kaplan-Meier (K-M) analyses indicated that the high-risk score in the train set (P < 0.0001), as well as the test set (P = 0.0053), corresponded with poorer survival. In addition, the high-risk score group showed a poor response to immunotherapy. The reliability and credibility of the risk scoring model were confirmed according to the CGGA dataset, TCGA datasets, and Pan-cancer analysis. According to drug sensitivity analysis, it was discovered that LJI308, a potent selective inhibitor of RSK pathways, has the highest drug sensitivity. Moreover, the GBM patients with higher risk scores may potentially be more beneficial from drugs that target cell cycle, mitosis, microtubule, DNA replication and apoptosis regulation signaling. CONCLUSION: We identified potential associations between clinical characteristics, TME, stemness, subtypes, and immunotherapy, and we clarified the therapeutic usefulness of cell senescence-associated genes.

Identification and validation of an anoikis-associated gene signature to predict clinical character, stemness, IDH mutation, and immune filtration in glioblastoma.

Background: Glioblastoma (GBM) is the most prominent and aggressive primary brain tumor in adults. Anoikis is a specific form of programmed cell death that plays a key role in tumor invasion and metastasis. The presence of anti-anoikis factors is associated with tumor aggressiveness and drug resistance. Methods: The non-negative matrix factorization algorithm was used for effective dimension reduction for integrated datasets. Differences in the tumor microenvironment (TME), stemness indices, and clinical characteristics between the two clusters were analyzed. Difference analysis, weighted gene coexpression network analysis (WGCNA), univariate Cox regression, and least absolute shrinkage and selection operator regression were leveraged to screen prognosis-related genes and construct a risk score model. Immunohistochemistry was performed to evaluate the expression of representative genes in clinical specimens. The relationship between the risk score and the TME, stemness, clinical traits, and immunotherapy response was assessed in GBM and pancancer. Results: Two definite clusters were identified on the basis of anoikis-related gene expression. Patients with GBM assigned to C1 were characterized by shortened overall survival, higher suppressive immune infiltration levels, and lower stemness indices. We further constructed a risk scoring model to quantify the regulatory patterns of anoikis-related genes. The higher risk score group was characterized by a poor prognosis, the infiltration of suppressive immune cells and a differentiated phenotype, whereas the lower risk score group exhibited the opposite effects. In addition, patients in the lower risk score group exhibited a higher frequency of isocitrate dehydrogenase (IDH) mutations and a more sensitive response to immunotherapy. Drug sensitivity analysis was performed, revealing that the higher risk group may benefit more from drugs targeting the PI3K/mTOR signaling pathway. Conclusion: We revealed potential relationships between anoikis-related genes and clinical features, TME, stemness, IDH mutation, and immunotherapy and elucidated their therapeutic value.

Effective Optical Zone Following Small Incision Lenticule Extraction for Myopia Calculated With Two Novel Methods.

PURPOSE: To evaluate the effective optical zone (EOZ) following small incision lenticule extraction in myopic eyes using two novel methods and investigate factors influencing postoperative EOZ. METHODS: In this prospective study, 45 patients (45 eyes) with a mean spherical equivalent of -5.82 ± 1.58 diopters underwent SMILE and were observed during a 6-month follow-up. Postoperative EOZ was calculated using custom software that automatically distinguishes EOZ on the tangential curvature difference map (EOZc) and total corneal refractive power map (EOZp) of the Pentacam HR (Oculus Optikgeräte GmbH). The agreement between the two methods, the difference between postoperative EOZ and programmed optical zone (POZ), and its relationship with parameters including corrected spheres, cylinders, ablation ratio (ablation depth/central cornea thickness), and Q-value change were investigated. RESULTS: The EOZc area was 20.76 ± 2.43 mm2 (diameter: 5.04 ± 0.60 mm) and the EOZp area was 20.22 ± 4.70 mm2 (diameter: 5.13 ± 0.30 mm). Both were significantly smaller than POZ (P < .001). Bland-Altman plots showed 4.44% (2/45) points located outside the 95% limits of agreement. EOZc and EOZp reductions were negatively related to corrected cylinders (r = -0.631, P < .001 and r = -0.594, P < .001, respectively). EOZp reduction was positively correlated with corrected spheres (r = 0.336, P = .024). Subgroup analysis revealed significant differences in EOZc and EOZp reduction between low and high astigmatism groups despite myopia degree. CONCLUSIONS: EOZ after incision lenticule extraction, measured using two novel methods, was smaller than POZ. The reduction of EOZ was negatively correlated with the corrected cylinders. [J Refract Surg. 2022;38(7):414-421.].

Lelliottia steviae sp. nov. isolated from Stevia rebaudiana Bertoni.

A Gram-negative, aerobic, chemoheterotrophic, rod-shaped, and motile bacterium, designated as LST-1T, was isolated from wild Stevia rebaudiana Bertoni and subjected to a polyphasic taxonomic analysis. The LST-1 strain grew optimally at 37 °C and pH 6.0-7.0 in the presence of 0.5% (w/v) NaCl. Phylogenetic analysis based on the 16S rDNA sequence indicated that LST-1 is closely related to Lelliottia jeotgali PFL01T (99.85%), Lelliottia nimipressuralis LMG10245T (98.82%), and Lelliottia amnigena LMG2784T (98.54%). Multi-locus sequence typing of concatenated partial atpD, infB, gyrB, and rpoB genes was performed to improve the resolution, and clear distinctions between the closest related type strains were observed. The results of average nucleotide identify analyses and DNA-DNA hybridization with four species (16S rDNA similarity > 98.65%) were less than 90 and 40%, respectively, verifying the distinct characteristics from other species of Lelliottia. The cellular fatty acid profile of the strain consisted of C16:0, Summed Feature3, and Summed Feature8 (possibly 16:1 w6c/16:1 w7c and 18:1 w6c) as major components. The major polar lipids included phosphatidylethanolamine, phosphatidylglycerol, an aminophospholipid, three non-characteristic phospholipids, and a non-characteristic lipid. The genome of LST-1T was 4,611,055 bp in size, with a G + C content of 55.02%. The unique combination of several phenotypic, chemotaxonomic, and genomic characteristics proved that strain LST-1T belongs to a novel species, for which the name Lelliottia steviae sp. nov. is proposed. The type strain is LST-1T (= CGMCC 1.19175T = JCM 34938T).Repositories: The genbank accession numbers for the 16S rRNA gene and genome sequences of strain LST-1T are MZ497264 and CP063663, respectively.

Characteristics of bile microbiota in cholelithiasis, perihilar cholangiocarcinoma, distal cholangiocarcinoma, and pancreatic cancer.

Significant proof suggests an essential role played by the bile microbiota in biliary diseases. This study retrospectively analyzed the differences in biliary microbes among patients with perihilar cholangiocarcinoma (pCCA), distal cholangiocarcinoma (dCCA), pancreatic cancer (PC), and cholelithiasis (CH). Bile samples were obtained from 53 patients who underwent endoscopic retrograde cholangiopancreatography (ERCP), and the bile microbiota was analyzed through 16S rRNA gene analysis and next-generation sequencing. Based on the results of linear discriminant analysis effect size (LEfSe), the top three biomarkers for pCCA at the genus level were Pseudomonas, Sphingomonas, and Halomonas; for dCCA were Streptococcus, Prevotella, and Halomonas; and for PC were Pseudomonas, Chloroplast, and Acinetobacter. The top five genera in the pCCA, dCCA, and PC groups showed predictive values with areas under the receiver operating characteristic curves of 91.56%, 95.56%, and 96.59%, respectively. The PICRUSt2 analysis outcomes displayed the diversities of fifteen pathways between the CH and pCCA groups, 22 pathways between the CH and dCCA groups, and eighteen pathways between the CH and PC groups. As this pilot study identified specific microbial bile markers for patients with CH, pCCA, dCCA, and PC, the clinical implications are vast. Further study focusing on distinct bacterial populations in bile will help differentiate biliary diseases.

Afatinib, an effective treatment for patient with lung squamous cell carcinoma harboring uncommon EGFR G719A and R776C co-mutations.

BACKGROUND: Epidermal growth factor receptor (EGFR) is a crucial driven gene in non-small cell lung cancer (NSCLC), and the EGFR mutation rate in lung squamous cell carcinoma (SCC) is only 3 ~ 6.92%. Uncommon EGFR mutations, such as S768I, L861Q and G719X, accounting for approximately 15% of NSCLC harboring EGFR mutation. Afatinib, a second-generation EGFR-tyrosine kinase inhibitor (TKI), has been approved for NSCLC harboring uncommon mutations by the FDA in 2018. In our report, the lung SCC patient harboring uncommon compound EGFR mutation (G719A and R776C) benefited from afatinib. CASE PRESENTATION: A case of a lung SCC patient harboring uncommon compound EGFR mutation (G719A and R776C) benefited from afatinib, and new MYC amplification was detected by next-generation sequencing (NGS) after disease progression. CONCLUSIONS: This case first identified a patient with lung squamous cell carcinoma harboring uncommon compound EGFR mutation (G719A and R776C) benefited from afatinib and achieved 11 months of progression-free survival (PFS). Then, new MYC amplification was detected after disease progression, indicating that MYC amplification may be one of the reasons for afatinib resistance.

Cerebrospinal fluid metabolic profiling reveals divergent modulation of pentose phosphate pathway by midazolam, propofol and dexmedetomidine in patients with subarachnoid hemorrhage: a cohort study.

BACKGROUND: Agitation is common in subarachnoid hemorrhage (SAH), and sedation with midazolam, propofol and dexmedetomidine is essential in agitation management. Previous research shows the tendency of dexmedetomidine and propofol in improving long-term outcome of SAH patients, whereas midazolam might be detrimental. Brain metabolism derangement after SAH might be interfered by sedatives. However, how sedatives work and whether the drugs interfere with patient outcome by altering cerebral metabolism is unclear, and the comprehensive view of how sedatives regulate brain metabolism remains to be elucidated. METHODS: For cerebrospinal fluid (CSF) and extracellular space of the brain exchange instantly, we performed a cohort study, applying CSF of SAH patients utilizing different sedatives or no sedation to metabolomics. Baseline CSF metabolome was corrected by selecting patients of the same SAH and agitation severity. CSF components were analyzed to identify the most affected metabolic pathways and sensitive biomarkers of each sedative. Markers might represent the outcome of the patients were also investigated. RESULTS: Pentose phosphate pathway was the most significantly interfered (upregulated) pathway in midazolam (p = 0.0000107, impact = 0.35348) and propofol (p = 0.00000000000746, impact = 0.41604) groups. On the contrary, dexmedetomidine decreased levels of sedoheptulose 7-phosphate (p = 0.002) and NADP (p = 0.024), and NADP is the key metabolite and regulator in pentose phosphate pathway. Midazolam additionally augmented purine synthesis (p = 0.00175, impact = 0.13481) and propofol enhanced pyrimidine synthesis (p = 0.000203, impact = 0.20046), whereas dexmedetomidine weakened pyrimidine synthesis (p = 0.000000000594, impact = 0.24922). Reduced guanosine diphosphate (AUC of ROC 0.857, 95%CI 0.617-1, p = 0.00506) was the significant CSF biomarker for midazolam, and uridine diphosphate glucose (AUC of ROC 0.877, 95%CI 0.631-1, p = 0.00980) for propofol, and succinyl-CoA (AUC of ROC 0.923, 95%CI 0.785-1, p = 0.000810) plus adenosine triphosphate (AUC of ROC 0.908, 95%CI 0.6921, p = 0.00315) for dexmedetomidine. Down-regulated CSF succinyl-CoA was also associated with favorable outcome (AUC of ROC 0.708, 95% CI: 0.524-0.865, p = 0.029333). CONCLUSION: Pentose phosphate pathway was a crucial target for sedatives which alter brain metabolism. Midazolam and propofol enhanced the pentose phosphate pathway and nucleotide synthesis in poor-grade SAH patients, as presented in the CSF. The situation of dexmedetomidine was the opposite. The divergent modulation of cerebral metabolism might further explain sedative pharmacology and how sedatives affect the outcome of SAH patients.

PPARα agonist fenofibrate relieves acquired resistance to gefitinib in non-small cell lung cancer by promoting apoptosis via PPARα/AMPK/AKT/FoxO1 pathway.

Recent studies show that intracellular accumulation of cholesterol leads to acquired resistance to gefitinib in non-small cell lung cancer (NSCLC) cells. In this study we investigated how to regulate the cholesterol levels in gefitinib-resistant NSCLC cells. We showed that intracellular cholesterol levels in gefitinib-resistant cell lines (PC-9/GR, H1975, H1650, and A549) were significantly higher than that in gefitinib-sensitive cell line (PC-9). Treatment with gefitinib (5 µM) significantly increased intracellular cholesterol levels in PC-9/GR, H1975, and H1650 cells. Gefitinib treatment downregulated the expression of PPARα, LXRα, and ABCA1, leading to dysregulation of cholesterol efflux pathway. We found that a lipid-lowering drug fenofibrate (20, 40 µM) dose-dependently increased the expression of PPARα, LXRα, and ABCA1, decreased the intracellular cholesterol levels, and enhanced the antiproliferative effects of gefitinib in PC-9/GR, H1975, and H1650 cells. We revealed that fenofibrate increased the gefitinib-induced apoptosis via regulating the key proteins involved in the intrinsic apoptosis pathway. In PC-9/GR, H1975 and H1650 cells, fenofibrate dose-dependently increased the expression of AMPK, FoxO1, and decreased the expression of AKT, which were remarkably weakened by knockdown of PPARα. In PC-9/GR cell xenograft mice, combined administration of gefitinib (25 mg · kg-1 · d-1) and fenofibrate (100 mg · kg-1 · d-1) caused remarkable inhibition on tumor growth as compared to treatment with either drug alone. All the results suggest that fenofibrate relieves acquired resistance to gefitinib in NSCLC by promoting apoptosis via regulating PPARα/AMPK/AKT/FoxO1 pathway. We propose that combination of gefitinib and fenofibrate is a potential strategy for overcoming the gefitinib resistance in NSCLC.