Analytical Modeling of Riveting Squeezing Force Considering Non-Uniform Deformation of Rivets in Aeronautical Structures.

Analytical modeling of the squeezing force for aircraft wings and fuselage panels in the existing literature usually assumes uniform deformation of the rivets, while in reality, the deformation of the rivets is non-uniform. To achieve high-quality squeezing force modeling, this paper introduces Coulomb's friction and four critical adjustments to the original equation: the non-uniform rivet/sheet interference along the sheet's hole axial ordinate; the barreling effect when calculating the driven head's volume; the spring-back of the driven head's dimensions; the modified height of the driven head; and the modified sheet-hole expanded diameter considering the convex structure of the driven head. The calculated values of the proposed new model demonstrate an improved level of accuracy, particularly under squeeze ratios commonly encountered in the aerospace industry.

Regulating Schwann Cell Growth by Nanosecond Pulsed Electric Field for Peripheral Nerve Regeneration In Vitro.

Schwann cells (SCs) are myelinating cells of the peripheral nervous system, playing a crucial role in peripheral nerve regeneration. Nanosecond Pulse Electric Field (nsPEF) is an emerging method applicable in nerve electrical stimulation that has been demonstrated to be effective in stimulating cell proliferation and other biological processes. Aiming to assess whether SCs undergo significant changes under nsPEF and help explore the potential for new peripheral nerve regeneration methods, cultured RSC96 cells were subjected to nsPEF stimulation at 5 kV and 10 kV, followed by continued cultivation for 3-4 days. Subsequently, some relevant factors expressed by SCs were assessed to demonstrate the successful stimulation, including the specific marker protein, neurotrophic factor, transcription factor, and myelination regulator. The representative results showed that nsPEF significantly enhanced the proliferation and migration of SCs and the ability to synthesize relevant factors that contribute positively to the regeneration of peripheral nerves. Simultaneously, lower expression of GFAP indicated the benign prognosis of peripheral nerve injuries. All these outcomes show that nsPEF has great potential as an efficient treatment method for peripheral nerve injuries by stimulating SCs.

A simple and sensitive ultra-high performance liquid chromatography tandem mass spectrometry method for the quantitative analysis of VX-548 in monkey plasma: Method validation and application to pharmacokinetic study.

VX-548 is an orally active and highly selective NaV 1.8 inhibitor that is undergoing development for the treatment of acute pain. The aim of this study was to develop a liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the measurement of VX-548 in monkey plasma. VX-548 was extracted from the plasma using acetonitrile-mediated protein precipitation. The quantitative analysis was performed on a Thermo Vantage TSQ mass spectrometer with ibrutinib as an internal standard. Chromatography was performed on a Waters ACQUITY UPLC BEH C18 column with 0.1% aqueous formic acid and acetonitrile as mobile phase. The precursor-to-product ion transitions were m/z 474.2 > 165.0 and m/z 441.2 > 138.1 for VX-548 and internal standard, respectively. This developed method was successfully validated in the concentration range of 1-1000 ng/mL. The calibration curve showed excellent linearity with a correlation coefficient of >0.999. The precision expressed as relative standard deviation (RSD) was <8.4%, whereas the accuracy denoted as relative error (RE) ranged from -5.0% to 9.1%. The mean recovery was >84%. VX-548 was stable in monkey plasma after storage under certain conditions. The validated method was successfully applied to the pharmacokinetic study of VX-548 in monkey plasma after single oral (2 mg/kg) and intravenous (1 mg/kg) administrations.

Relationship between resting heart rate and long-term outcomes in stabilized patients with myocardial infarction: A prospective community-based cohort study.

BACKGROUND: Resting heart rate (RHR) during hospitalization has been shown to be associated with adverse outcomes in patients with myocardial infarction (MI). This study aimed to evaluate the long-term prognostic effect of RHR during the stable phase after MI in post-MI patients. METHODS: Patients who had prior or new-onset MI and RHR measurements during the stable period after MI between 2006 and 2018 in the community-based Kailuan Study were enrolled. RHR was divided into four groups based on quartiles. Cox regression analysis was used to analyze the association of RHR with primary composite outcome of all-cause death, hospitalization for heart failure (HF), stroke, and recurrent MI and its components. RESULTS: A total of 4447 post-MI patients were included. During a median follow-up of 7.5 years, 1813 patients (40.8%) developed primary outcomes. Compared to RHR ≤67 bpm, patients with 72 < RHR ≤80 bpm and RHR >80 bpm had increased risks of primary outcome, with adjusted hazard ratios (95% confidence intervals) of 1.23 (1.08-1.40) and 1.35 (1.18-1.55), respectively. The risk of primary outcome increased by 12% (1.07-1.17) for each 10-bpm increase in RHR. Similar results were observed in all-cause death and hospitalization for HF. Restricted cubic splines revealed a linear relationship between RHR and primary outcome, all-cause death, and hospitalization for HF (P for nonlinearity >0.05). CONCLUSIONS: RHR during the stable phase after MI was an independent predictor for primary outcome and all-cause death in post-MI patients, and RHR >72 bpm was associated with increased risk for primary outcome and all-cause death.

Influence of Pre-Heat Treatment on the Deformation Behaviors, Microstructural Characteristics, and Mechanical Properties of a Continuously Cast Al-Cu-Mg Alloy during Continuous Extrusion Process.

The presence of a second phase in Al-Cu-MG alloys, with various sizes and supersaturation-solid-solubility, which can be changed by pre-heat-treatment, could have remarkable influence on hot workability and mechanical performance. In the present work, a continuously cast 2024 Al alloy was homogenized and then subjected to hot compression and continuous extrusion (Conform) along with the initial as-cast alloy. The results showed that the 2024 Al alloy specimen with pre-heat treatment had a higher resistance to deformation and dynamic recovery (DRV) during hot compression process compared with the as-cast sample. Meanwhile, dynamic recrystallization (DRX) was advanced in the pre-heat-treated sample. After the Conform Process, the pre-heat-treated sample also attained better mechanical properties without additional solid solution treatment. The higher supersaturation solid solubility and dispersoids generated during pre-heat treatment was proved to play a key role in restricting boundary migration, tangling dislocation motion and promoting the precipitation of S phase, which raised resistance to DRV and plastic deformation and enhanced the mechanical properties.

Correction: Noncovalent wedging effect catalyzed the cis to syn transformation of a surface-adsorbed polymer backbone toward an unusual thermodynamically stable supramolecular product.

Correction for 'Noncovalent wedging effect catalyzed the cis to syn transformation of a surface-adsorbed polymer backbone toward an unusual thermodynamically stable supramolecular product' by Zhi-Xuan Liu et al., Phys. Chem. Chem. Phys., 2022, 24, 30010-30016, https://doi.org/10.1039/D2CP04184G.

New advances in treatment of skin malignant tumors with nanosecond pulsed electric field: A literature review.

BACKGROUND: Nanosecond pulsed electric field, with its unique bioelectric effect, has shown broad application potential in the field of tumor therapy, especially in malignant tumors and skin tumors. MAIN BODY: In this paper, we discuss the therapeutic effects and mechanisms of nanosecond pulsed electric field on three common skin cancers, namely, malignant melanoma, squamous cell carcinoma and basal cell carcinoma, as well as its application to other benign skin diseases and future development and improvement directions. CONCLUSION: In general, nanosecond pulsed electric field mainly exerts its ablative effect on tumors through subcellular membrane electroporation effect. It is cell type-specific, has less thermal damage, and can have synergistic effect with chemotherapy drugs, making it a very promising new method for tumor treatment.

Construction and Validation of a Protein-associated Prognostic Model for Gastrointestinal Cancer.

Background Gastrointestinal cancer (GIC) is a prevalent and lethal malignant tumor. It is obligatory to investigate innovative biomarkers for the diagnosis and prognosis. Proteins play a crucial role in regulating the occurrence and progression of GIC. However, the prognostic value of proteins is unclear in GIC. OBJECTIVE: This paper aims to identify the hub prognosis-related proteins (PAPs) and construct a prognosis model for GIC patients for clinical application. METHODS: Protein expression data of GIC was obtained from The Cancer Proteome Atlas (TCPA) and downloaded the clinicopathological data from The Cancer Genome Atlas database (TCGA). Besides, hub proteins were filtrated via univariate and multivariate Cox regression analysis. Moreover, survival analysis and nomogram were used to predict overall survival (OS). We used the calibration curves to assess the consistency of predictive and actual survival rates. The consistency index (C-index) was used to evaluate the prognostic ability of the predictive model. Furthermore, functional enrichment analysis and protein co-expression of PAPs were used to explore their roles in GIC. RESULTS: Finally, a prognosis model was conducted based on ten PAPs (CYCLIND1, DVL3, NCADHERIN, SYK, ANNEXIN VII, CD20, CMET, RB, TFRC, and PREX1). The risk score calculated by the model was an independent prognostic predictor. Compared with the high-risk subgroup, the low-risk subgroup had better OS. In the TCGA cohort, the area under the curve value of the receiver operating characteristic curve of the prognostic model was 0.692. The expression of proteins and risk score had a significant association with the clinicopathological characteristics of GIC. Besides, a nomogram based on GIC clinicopathological features and risk scores could properly predict the OS of individual GIC patients. The C-index is 0.71 in the TCGA cohort and 0.73 in the GEO cohort. CONCLUSION: The results indicate that the risk score is an independent prognostic biomarker and is related to the malignant clinical features of GIC patients. Besides, several PAPs associated with the survival and clinicopathological characteristics of GIC might be potential biomarkers for GIC diagnosis and treatment.

UPLC-Q-TOF-MS/MS analysis on the chemical composition of malts under different germination cycles and prepared with different processing methods.

OBJECTIVE: To investigate changes in the chemical composition of malts under different germination cycles and prepared with different processing methods, thus providing a reference for the clinical application of malt in disease treatment. METHODS: Nine malt samples were analyzed by ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS), and the MS fragmentation pathway of 4 compounds (including hordenine, gramine, N-methyltyramine and catechin) were also analyzed. RESULTS: By database comparison and literature search, we detected 31 compounds in raw barley and 33 compounds in both raw malt and roasted malt. Nonetheless, the most of these 33 compounds were detected higher contents in raw malt than in roasted malt. Besides, we detected 15 compounds in brown malt. At Day1 of germination, 31 compounds were detected in malt, without two alkaloids (representative: hordenine). At Day2-5, 33 compounds were detected, with different contents as shown by the peak area comparison; hordenine had a gradually increasing abundance; and nearly one third of the chemical components in barley increased gradually, one third decreased gradually, and one third tended to be stable. CONCLUSION: Malts under different germination cycles and prepared with different processing methods have varying active ingredients, and especially brown malt exhibits a serious loss of compounds. The tight association between the chemical composition and clinical application of malt offers a basis to the clinically scientific and reasonable selection of Chinese medicinal materials for treatment purposes.

Improved Insulating Properties of Polymer Dielectric by Constructing Interfacial Composite Coatings.

Polymeric dielectrics exhibit remarkable dielectric characteristics and wide applicability, rendering them extensively employed within the domain of electrical insulation. Nevertheless, the electrical strength has always been a bottleneck, preventing its further utilization. Nanocomposite materials can effectively improve insulation strength, but uniform doping of nanofillers in engineering applications is a challenge. Consequently, a nanocomposite interfacial coating was meticulously designed to interpose between the electrode and the polymer, which can significantly improve DC breakdown performance. Subsequently, the effects of filler concentration and coating duration on DC breakdown performance, high field conductivity, and trap distribution characteristics were analyzed. The results indicate that the composite coating introduces deep traps between the electrode-polymer interface, which enhances the carrier confinement, resulting in reduced conductivity and enhanced DC breakdown strength. The incorporation of a composite coating at the interface between the electrode and polymer presents novel avenues for enhancing the dielectric insulation of polymers.

Noncovalent wedging effect catalyzed the cis to syn transformation of a surface-adsorbed polymer backbone toward an unusual thermodynamically stable supramolecular product.

The significant influence of noncovalent interactions on catalytic processes has been recently appreciated but is still in its infancy. In this report, it is found that wedging Me-PTCDI (small-molecule) between the alkyl chains of PffBT4T-2OD (polymer) and a graphite substrate can reduce the energy barrier of flipping over the surface-adsorbed alkylthiophene group from the cis to syn conformation, revealing the catalytic role of Me-PTCDI via a noncovalent wedging effect. The wedging of Me-PTCDI brings the interactions between the alkyl chains and substrate to a very weak level by lifting up the alkyl chains, which eliminates the major hindrance of the flipping process to one main factor: the torsion of the dihedral angles of the thiophene group. The Me-PTCDI/syn PffBT4T-2OD arrangement shows unusual stability compared to the cis one because the syn conformation allows the alkyl chains to construct dense lamella and facilitates interactions between Me-PTCDI and the syn PffBT4T-2OD backbones. The results are helpful for boosting the development of noncovalent catalysis and bottom-up fabrications toward devices functionalized at a molecular level.

Pulsed-Field Ablation Using a Novel Ablation-Mapping Integrated System for Pulmonary Vein Isolation-A Preliminary Animal Study.

OBJECTIVE: The purpose of this study is to evaluate the preliminary safety and effect of a pulsed electric field (PEF) ablation system. METHODS: The pulmonary veins (PVs) and superior vena cava (SVC) were isolated with the pulsed field ablation (PFA) system, which included a PEF generator and an electrode. The effects of PFA were investigated in six porcines using a novel circular catheter with combined functions (mapping/ablation) designed to work with a cardiac mapping system. The PEF generator delivered a train of biphasic pulsed electric pulses with a high amplitude (800-2000 V) and short pulse duration. The voltage mapping, PVs and SVC potentials, ostial diameters, and phrenic nerve and esophagus viability data were collected 4 weeks later, after which the animals were subsequently euthanized for gross histopathology analysis. RESULTS: PFA 100% isolated the PVs and SVC with four applications with a mean pulse number of 100-150 pulses, causing no muscle convulsion. PFA does not cause PV stenosis or phrenic nerve dysfunction. Histological analysis confirmed 100% transmurally without any venous stenoses or phrenic injuries. Pathology follow-up showed that PFA had selectively ablated cardiomyocytes but spared blood vessels, the esophagus, and phrenic nerves; after ablation, the myocardial tissue showed homogeneous fibrosis. CONCLUSION: The PFA system is safe and feasible in the preliminary porcine model, which can effectively isolate PVs and SVCs. Transmural tissue damage can be achieved without phrenic palsy or stenosis.

Study of necrotic apoptosis by pulsed electric field ablation in rabbit left ventricular myocardium.

Objective: We investigate the characteristics of histological damage to myocardial cells in the ablation region and surrounding areas of the left ventricular epicardium in rabbits using our self-developed cardiac pulsed electric field (PEF) ablation instrument and ablation catheter. Methods: Forty eight New Zealand rabbits underwent ablation on the left ventricular myocardium after open-heart exposure with a cardiac arrhythmia PEF ablation device and ablation catheter developed by the Medical Translation Laboratory of Pulsed Electric Field Technology in Zhejiang Province. The ablation parameters were set as biphasic electrical pulses; voltage, ±800 V; pulse width, 10 µs; interphase delay, 500 us. Six rabbits were included in the sham group and 42 other rabbits were randomly divided into immediately, 6-h, 1-, 3-day, 1-, 2-, and 4-week post-ablation groups, with six rabbits in each group. Creatine kinase- (CK)-MB isoenzyme (CK-MB), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) levels were measured before and at different time points after PEF ablation to analyze their dynamic evolution. Masson staining of tissue block sections of left ventricular myocardial ablation and adjacent tissue heart specimens was performed, and the occurrence of TUNEL apoptosis in myocardium tissue was analyzed. Results: All rabbits completed the PEF ablation procedure and the follow-up process. After PEF ablation, the levels of cardiac enzymes, including CK-MB, CK, and AST, increased significantly, peaking 1-3 days after the procedure. In particular, those of CK and CK-MB increased by 15-20 times but returned to the preoperative level after 2 weeks. Based on general observation, it was found that the myocardium in the ablation area was swollen immediately after PEF ablation. Masson staining analysis revealed that cardiomyocytes were broken and infiltrated by erythrocytes after 6 h. After 1 day, the cells started to experience atrophy and necrosis; after 3 days, fibrotic replacement of the necrotic area became obvious. Then, by 4 weeks, the myocardial cells were completely replaced by hyperplasia. Apoptosis occurred significantly at 6 h and peaked at 24 h post-ablation, demonstrating a 37.7-fold increase; apoptotic cell counts decreased significantly at 3 days post-ablation, and no significant apoptotic cardiomyocytes were seen after 1 week. Conclusion: After PEF ablation, cardiomyocytes showed apoptotic process and dyed, at least partially, through a secondary necrosis, the ablation boundary was clear, the ablation area was replaced by structurally intact fibroblasts, no island myocardium tissue were seen, and the ablation area vessels and nerves were not affected.

TLR4 inhibition suppresses growth in oestrogen-induced prolactinoma models.

Prolactinomas have harmful effects on human health. Bromocriptine is the only commercially available drug in China, but about 25% of prolactinoma patients do not respond to it in clinic, its pathogenesis remains unknown. Thus, its pathogenesis needs to be determined to develop new therapeutic methods for prolactinomas. The expression of ERß, TLR4, and prolactin (PRL) in the pituitary gland of C57BL/6 mice and human prolactinoma specimen was examined by immunofluorescence or immunohistochemistry. The role of TLR4 in prolactinoma was determined using estradiol-induced models of C57BL/6 wild-type and TLR4-/- mice. MMQ cells were treated with estradiol, fulvestrant, and lipopolysaccharide (LPS) or transfected with TLR4 siRNA to study the expression of ERß, TLR4, and PRL in these cells. Furthermore, the interaction between ERß and TLR4 was investigated by immunoprecipitation analysis. The expression of PRL and TLR4 was co-located and increased in the pituitary gland of mice and human prolactinoma specimen compared to that in the control specimen. Meanwhile, TLR4 knockout or treatment with the TLR4 inhibitor TAK242 not only significantly inhibited tumor overgrowth but also decreased the expression of PRL in estradiol-treated mice through p38 MAPK pathway regulation. However, MMQ treated with estradiol and LPS enhanced PRL expression than treated with estradiol or LPS alone. Finally, ERß or TLR4 inhibition prevented the estradiol-induced PRL increase by regulating the TLR4/p38 MAPK pathway in vitro. Estradiol promoted prolactinoma development by activating the TLR4/p38 MAPK pathway through ERß, and TLR4 is a potential therapeutic target for prolactinoma treatment.

Role of gasdermin family proteins in the occurrence and progression of hepatocellular carcinoma.

Primary liver cancer is the sixth most common cancer and the third leading cause of cancer mortality worldwide, hepatocellular carcinoma (HCC) is the most common type of liver cancer, accounting for 75%-85% of cases. The occurrence and progression of HCC involve multiple events. Pyroptosis is a gasdermins mediated programmed cell death and is intricately associated with cancerogenesis, including HCC. This review mainly concerns the recent research advances of the gasdermin family members in HCC. The biological roles and specific expression patterns of the family members are discussed, especially those that are involved in the regulatory pathways in the occurrence and progression of HCC. We provide the latest progress into the distinct molecular mechanisms of gasdermin family members involved in the occurrence and development of HCC.

Diagnostic Value of Color Doppler Ultrasonography in Subacute Thyroiditis.

In order to explore the clinical effect of color Doppler ultrasonography in the diagnosis of subacute thyroiditis, a method for the diagnosis of subacute thyroiditis by color Doppler ultrasonography was proposed. From November 2019 to November 2020, 90 patients with subacute thyroiditis in our hospital were selected as the experimental group; 90 healthy people were selected as the control group during the same period. Both groups were diagnosed by color Doppler ultrasonography and compared. The experimental results showed that patients with subacute thyroiditis showed mild to moderate enlargement of the involved thyroid gland, and local or diffuse inhomogeneous hypoechoic areas may appear in bilateral or unilateral thyroid glands: irregular edges, unclear boundaries, no "ball feel," mottled changes, and accompanied by tenderness. The blood flow signal around the hypoechoic area is rich, and the internal blood flow signal is less. There was no significant increase in the blood flow velocity of the superior thyroid artery on the affected side. Color Doppler ultrasound not only is simple, economical, and non-invasive but also has a good diagnostic accuracy for subacute thyroiditis, which can provide an important basis for clinical diagnosis and treatment and is worthy of popularization and application.

Total Barley Maiya Alkaloids Prevent Increased Prolactin Levels Caused by Antipsychotic Drugs and Reduce Dopamine Receptor D2 via Epigenetic Mechanisms.

Background: The dopamine D2 receptor (DRD2) plays an important role in the increased prolactin (PRL) levels associated with the pathogenesis of antipsychotic drugs (ADs). Elevated prolactin levels can affect people's quality of life. Maiya alkaloids has been used to treat diseases associated with high PRL levels. Maiya, is a processed product of the mature fruits of Hordeum vulgare L. (a gramineous plant) after sprouting and drying and also a common Chinese herbal drug used in the clinic, is traditionally used to treat abnormal lactation, and is currently used clinically for the treatment of abnormal PRL levels. Aims: Epigenetic mechanisms can be related to DRD2 expression. We investigated the role of DRD2 methylation in the induction of PRL expression by ADs and the mechanism underlying the effects of total barley maiya alkaloids (TBMA) on this induction. Methods: The methylation rate of DRD2 in 46 people with schizophrenia who took risperidone was detected by MassARRAY sequencing. Humans were long term users of Ris. Seventy Sprague Dawley female rats were divided into seven groups. A rat model of risperidone-induced PRL was established, and the potential protective effects of TBMA and its components [e.g., hordenine (Hor)] on these increased PRL levels were investigated. The PRL concentration was detected by Enzyme-linked immunosorbent assay. PRL, DRD2, and DNA methyltransferase (DNMT1, DNMT3α, and DNMT3ß) protein and mRNA expression were detected by western blotting and real-time polymerase chain reaction (RT-PCR), respectively. The positive rate of methylation in the DRD2 promoter region of rats was detected by MassARRAY sequencing. Results: Clinical studies showed that the positive rate of DRD2 methylation associated with increased PRL levels induced by ADs was significantly higher than in the normal prolactinemia (NPRL) group. In vivo and vitro, TBMA and Hor inhibited this induction of PRL expression and increased DRD2 expression by inhibiting the expression of the DNMTs. Conclusions: TBMA and hordenine increased DRD2 expression by inhibiting DNMT-dependent DRD2 methylation.

Modification of benzoindenothiophene-based organic dye with fused thiophenes for efficient dye-sensitized solar cells.

Recently low-energy-gap benzoindenothiophene (BIT)-based organic dyes have been experimentally sensitized to dye-sensitized solar cells (DSSCs) with impressive 10.9% power conversion efficiency. This paper presents a computational study of the modification of BIT-based dyes with fused thiophene moieties to build novel low band gap sensitizers. Density functional theory (DFT), tight-binding DFT, and time dependent DFT (TDDFT) approaches are used to demonstrate the electronic and optical properties of the BIT dyes and dye/(TiO2)46 complexes. Our calculations show that the structural modification by using fused thiophenes can effectively lower the band gap of the BIT dyes by 0.07-0.12 eV and affect the optical properties of BIT dyes. Enlarging the thiophene unit in BIT with thienothiophene and dithienothiophene improves the oscillator strength by 14%-25%, while the lowest-energy absorption peak basically remains at 559 nm. The incorporation of cyclopentadithiophene unit leads to a significant 47 nm red-shift of absorption peak and a 25% enhanced oscillator strength, compared to the original BIT dye. Those fused thiophenes modified BIT dyes also demonstrate ideal molecular orbital distribution patterns and ultra-fast injection time at the dye/(TiO2)46 interface. Our calculations provide useful guidance for the molecular design of novel naphthalene-based dyes for DSSC optimizations.

Construction and validation of m6 A RNA methylation regulators associated prognostic model for gastrointestinal cancer.

N6-methyladenosine (m6 A) RNA methylation is correlated with carcinogenesis and dynamically possessed through the m6 A RNA methylation regulators. This paper aimed to explore 13 m6 A RNA methylation regulators' role in gastrointestinal cancer (GIC) and determine the risk model and prognosis value of m6 A RNA methylation regulators in GIC. We used several bioinformatics methods to identify the differential expression of m6 A RNA methylation regulators in GIC, constructed a prognostic model, and carried out functional enrichment analysis. Eleven of 13 m6 A RNA methylation regulators were differentially expressed in different clinicopathological characteristics of GIC, and m6 A RNA methylation regulators were nearly associated with GIC. We constructed a risk model based on five m6 A RNA methylation regulators (METTL3, FTO, YTHDF1, ZC3H13, and WTAP); the risk score is an independent prognosis biomarker. Moreover, the five m6 A RNA methylation regulators can also forecast the 1-, 3- and 5-year overall survival through a nomogram. Furthermore, four hallmarks of oxidative phosphorylation, glycolysis, fatty acid metabolism, and cholesterol homoeostasis gene sets were significantly enriched in GIC. m6 A RNA methylation regulators were related to the malignant clinicopathological characteristics of GIC and may be used for prognostic stratification and development of therapeutic strategies.

Cytochrome P450 3A5 polymorphism affects the metabolism of sorafenib and its toxicity for hepatocellular carcinoma cells in vitro.

OBJECTIVE: Cytochrome P450 3A5 (CYP3A5) is a highly polymorphic gene and the encoded protein variants differ in catalytic activity, leading to inter-individual variation in metabolic ability. The aim of the current study was to investigate the effects of seven allelic variants on the ability of CYP3A5 to metabolize sorafenib in vitro and further explore the impacts of CYP3A5 polymorphism on the proliferation and apoptosis of hepatocellular carcinoma cell line (HepG2) induced by sorafenib. METHODS: Wild-type and variant CYP3A5 enzymes were expressed in Spodoptera frugiperda insect cells using a baculovirus dual-expression system, and protein expression was checked by western blot. The enzymes were incubated with sorafenib at 37°C for 30 min, and formation of the major metabolite sorafenib N-oxide was assayed using ultra-performance liquid chromatography and tandem mass spectrometry. Intrinsic clearance values (Vmax/Km) were calculated for each enzyme. Additionally, recombinant HepG2 cells transfecting with CYP3A5 variants were used to investigate the effects of sorafenib on the proliferation of HepG2 cells. RESULTS: Intrinsic clearance of the six variants CYP3A5*2, CYP3A5*3A, CYP3A5*3C, CYP3A5*4, CYP3A5*5, and CYP3A5*7 was 26.41-71.04% of the wild-type (CYP3A5*1) value. In contrast, the clearance value of the variant CYP3A5*6 was significantly higher (174.74%). Additionally, the decreased ATP levels and cell viability and the increased cell apoptosis in HepG2 cells transfected with CYP3A5*2, CYP3A5*3A, CYP3A5*3C, CYP3A5*4, CYP3A5*5, and CYP3A5*7 were observed, whereas, the increased ATP levels and cell viability and the reduced cell apoptosis in HepG2 cells transfected with CYP3A5*6 were also investigated when compared to CYP3A5*1. CONCLUSION: Our results suggest that CYP3A5 polymorphism influences sorafenib metabolism and pharmacotherapeutic effect in hepatic carcinomas. These data may help explain differential response to drug therapy for hepatocellular carcinoma, and they support the need for individualized treatment.