Exploring the Molecular Therapeutic Mechanisms of Gemcitabine through Quantitative Proteomics.

Gemcitabine (GEM) is widely employed in the treatment of various cancers, including pancreatic cancer. Despite their clinical success, challenges related to GEM resistance and toxicity persist. Therefore, a deeper understanding of its intracellular mechanisms and potential targets is urgently needed. In this study, through mass spectrometry analysis in data-dependent acquisition mode, we carried out quantitative proteomics (three independent replications) and thermal proteome profiling (TPP, two independent replications) on MIA PaCa-2 cells to explore the effects of GEM. Our proteomic analysis revealed that GEM led to the upregulation of the cell cycle and DNA replication proteins. Notably, we observed the upregulation of S-phase kinase-associated protein 2 (SKP2), a cell cycle and chemoresistance regulator. Combining SKP2 inhibition with GEM showed synergistic effects, suggesting SKP2 as a potential target for enhancing the GEM sensitivity. Through TPP, we pinpointed four potential GEM binding targets implicated in tumor development, including in breast and liver cancers, underscoring GEM's broad-spectrum antitumor capabilities. These findings provide valuable insights into GEM's molecular mechanisms and offer potential targets for improving treatment efficacy.

A Comprehensive Study of NF3-Based Selective Etching Processes: Application to the Fabrication of Vertically Stacked Horizontal Gate-All-around Si Nanosheet Transistors.

In this paper, we demonstrate a comprehensive study of NF3-based selective etching processes for inner spacer formation and for channel release, enabling stacked horizontal gate-all-around Si nanosheet transistor architectures. A cyclic etching process consisting of an oxidation treatment step and an etching step is proposed and used for SiGe selective etching. The cyclic etching process exhibits a slower etching rate and higher etching selectivity compared to the direct etching process. The cycle etching process consisting of Recipe 1, which has a SiGe etching rate of 0.98 nm/cycle, is used for the cavity etch. The process achieved good interlayer uniformity of cavity depth (cavity depth ≤ 5 ± 0.3 nm), while also obtaining a near-ideal rectangular SiGe etch front shape (inner spacer shape = 0.84) and little Si loss (0.44 nm@ each side). The cycle etching process consisting of Recipe 4 with extremely high etching selectivity is used for channel release. The process realizes the channel release of nanosheets with a multi-width from 30 nm to 80 nm with little Si loss. In addition, a selective isotropic etching process using NF3/O2/Ar gas mixture is used to etch back the SiN film. The impact of the O2/NF3 ratio on the etching selectivity of SiN to Si and the surface roughness of SiN after etching is investigated. With the introduction of O2 into NF3/Ar discharge, the selectivity increases sharply, but when the ratio of O2/NF3 is up to 1.0, the selectivity tends to a constant value and the surface roughness of SiN increases rapidly. The optimal parameter is O2/NF3 = 0.5, resulting in a selectivity of 5.4 and a roughness of 0.19 nm.

Self-protecting CoFeAl-layered double hydroxides enable stable and efficient brine oxidation at 2 A cm-2.

Low-energy consumption seawater electrolysis at high current density is an effective way for hydrogen production, however the continuous feeding of seawater may result in the accumulation of Cl-, leading to severe anode poisoning and corrosion, thereby compromising the activity and stability. Herein, CoFeAl layered double hydroxide anodes with excellent oxygen evolution reaction activity are synthesized and delivered stable catalytic performance for 350 hours at 2 A cm-2 in the presence of 6-fold concentrated seawater. Comprehensive analysis reveals that the Al3+ ions in electrode are etched off by OH- during oxygen evolution reaction process, resulting in M3+ vacancies that boost oxygen evolution reaction activity. Additionally, the self-originated Al(OH)n- is found to adsorb on the anode surface to improve stability. An electrode assembly based on a micropore membrane and CoFeAl layered double hydroxide electrodes operates continuously for 500 hours at 1 A cm-2, demonstrating their feasibility in brine electrolysis.

The MdNAC72-MdABI5 module acts as an interface integrating jasmonic acid and gibberellin signals and undergoes ubiquitination-dependent degradation regulated by MdSINA2 in apple.

Jasmonic acid (JA) and gibberellin (GA) coordinately regulate plant developmental programs and environmental cue responses. However, the fine regulatory network of the cross-interaction between JA and GA remains largely elusive. In this study, we demonstrate that MdNAC72 together with MdABI5 positively regulates anthocyanin biosynthesis through an exquisite MdNAC72-MdABI5-MdbHLH3 transcriptional cascade in apple. MdNAC72 interacts with MdABI5 to promote the transcriptional activation of MdABI5 on its target gene MdbHLH3 and directly activates the transcription of MdABI5. The MdNAC72-MdABI5 module regulates the integration of JA and GA signals in anthocyanin biosynthesis by combining with JA repressor MdJAZ2 and GA repressor MdRGL2a. MdJAZ2 disrupts the MdNAC72-MdABI5 interaction and attenuates the transcriptional activation of MdABI5 by MdNAC72. MdRGL2a sequesters MdJAZ2 from the MdJAZ2-MdNAC72 protein complex, leading to the release of MdNAC72. The E3 ubiquitin ligase MdSINA2 is responsive to JA and GA signals and promotes ubiquitination-dependent degradation of MdNAC72. The MdNAC72-MdABI5 interface fine-regulates the integration of JA and GA signals at the transcriptional and posttranslational levels by combining MdJAZ2, MdRGL2a, and MdSINA2. In summary, our findings elucidate the fine regulatory network connecting JA and GA signals with MdNAC72-MdABI5 as the core in apple.

Total Synthesis of (±)-Rubriflordilactone A.

A new and efficient synthesis of rubriflordilactone A has been realized. The key transformations include the following: (1) an intramolecular Prins cyclization to establish the seven-membered ring containing two contiguous stereocenters; (2) a Mukaiyama hydration/oxa-Michael cascade to construct the B-ring; and (3) an unprecedented stereocontrol intermolecular o-QM type [4 + 2]-cycloaddition to rapidly assemble core structure of rubriflordilactone A.

miR-125b-5p-MAPK1-C/EBPα feedback loop regulates all-trans retinoic acid resistance in acute promyelocytic leukemia.

BACKGROUND: Various studies have highlighted the significance of miR-125b-5p in tumour chemotherapy resistance; However, whether miR-125b-5p is associated with all-trans retinoic acid (ATRA) resistance in acute promyelocytic leukemia (APL) has not been reported. METHODS: Drug-resistance-related factors in APL were predicted using the DRESIS database. The expression levels of miR-125b-5p in ATRA-sensitive and ATRA-resistant APL cells were determined using quantitative reverse transcription polymerase chain reaction (qRT-PCR). A nitrotetrazolium blue (NBT) reduction assay and flow cytometry (FCM) were used to detect the effect of miR-125b-5p on ATRA resistance in APL cells. An APL xenograft tumour mouse model was established to determine the effect of miR-125b-5p on ATRA resistance. A dual-luciferase gene reporter assay, qRT-PCR, and western blotting verified the regulation by miR-125b-5p of its target gene, MAPK1, and the MAPK1 downstream factor, C/EBPα. An NBT reduction assay and FCM were used to detect the effect of C/EBPα on ATRA resistance in APL cells. Western blotting and qRT-PCR were used to assess the regulation of miR-125b-5p and MAPK1 by C/EBPα. RESULTS: miR-125b-5p expression levels were dramatically increased in ATRA-resistant APL cells. Both in vitro and in vivo experiments revealed that miR-125b-5p overexpression enhanced ATRA resistance in APL. miR-125b-5p promoted ATRA resistance by sponging MAPK1. C/EBPα was negatively regulated by miR-125b-5p, which in addition, regulated ATRA resistance in APL cells. C/EBPα also regulated the miR-125b-5p-MAPK1 axis. CONCLUSION: The findings of this study indicate that the miR-125b-5p-MAPK1-C/EBPα feedback loop regulated ATRA resistance in APL. Thus, miR-125b-5p may be a promising target for treating ATRA resistance in APL.

Aristolocholic acid I promotes renal tubular epithelial fibrosis by upregulating matrix metalloproteinase-9 expression via activating the C3a/C3aR axis of macrophages.

Aristolochic acid I (AAI) can cause nephrotoxicity and is characterized by interstitial fibrosis. The C3a/C3aR axis of macrophages and matrix metalloproteinase-9 (MMP-9) play important roles in fibrosis, but whether they are involved in AAI-induced renal interstitial fibrosis and are related remains to be elucidated. In this study, we investigated whether C3a/C3aR axis of macrophages promotes renal interstitial fibrosis by regulating MMP-9 in aristolochic acid nephropathy (AAN). Intraperitoneal injection of AAI for 28 days successfully induced AAN in C57bl/6 mice. The content of C3a in the kidney of AAN mice was increased, and there was a significant distribution of macrophages in the renal tubules. The same results were observed in the in vitro experiment. We also explored the role and mechanism of macrophages after AAI administration in the epithelial-mesenchymal transformation (EMT) of renal tubular epithelial cells (RTECs) and found that AAI could activate the C3a/C3aR axis of macrophages to upregulate p65 expression in macrophages. p65 upregulated MMP-9 expression in macrophages not only directly but also by promoting the secretion if interleukin-6 by macrophages and then activating STAT3 in RTECs. The upregulation of MMP-9 expression could promote the EMT of RTECs. Taken together, our study demonstrated that the AAI-activated the C3a/C3aR axis of macrophages, which induced MMP-9 production, was one of the causes of renal interstitial fibrosis. Therefore, targeting the C3a/C3aR axis of macrophages is an effective therapeutic strategy for the prevention and treatment of renal interstitial fibrosis in AAN.

Microstructure and Crystallization Kinetics of Silica-Based Ceramic Cores with Enhanced High-Temperature Property.

Silica-based ceramic cores play key roles in the casting of aeroengine blades, but they are highly limited by the poor high-temperature mechanical property. Here, fused mullite (FM) and sintered mullite (SM) powders were modified in silica-based ceramic cores, and the microstructure evolution and crystallization kinetics of ceramic cores depending on mullite types were studied. The ceramic cores with FM showed a dense microstructure and superior mechanical properties compared to those with SM. The ceramic cores with 10 wt.% of FM showed a crystallization activation energy of 1119.5 kJ/mol and a crystallization exponent of 1.74, and the values of 938.4 kJ/mol and 1.86 as SM were employed; the decreased crystallization activation energy and the elevated crystallization exponent by SM suggested that the excess impurities of alkali oxides and alkaline-earth oxides significantly promoted the crystallization of cristobalite. Even though the ceramic cores with mullite powders decreased slightly in the room-temperature mechanical property, their high-temperature flexure strength and creep deformation resistance were enhanced. The ceramic cores with 10 wt.% of FM showed excellent comprehensive performance, with linear shrinkage of 0.69%, room-temperature strength of 18.9 MPa, and high-temperature strength of 15.5 MPa, which satisfied the demands for hollow-blade casting.

Cytochrome P450 gene CYP6BQ8 mediates terpinen-4-ol susceptibility in the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae).

Cytochrome P450 proteins (CYPs) in insects can encode various detoxification enzymes and catabolize heterologous substances, conferring tolerance to insecticides. This study describes the identification of a P450 gene (CYP6BQ8) from Tribolium castaneum (Herbst) and investigation of its spatiotemporal expression profile and potential role in the detoxification of terpinen-4-ol, a component of plant essential oils. The developmental expression profile showed that TcCYP6BQ8 expression was relatively higher in early- and late-larval stages of T. castaneum compared with other developmental stages. Tissue expression profiles showed that TcCYP6BQ8 was mainly expressed in the head and integument of both larvae and adults. The expression profiling of TcCYP6BQ8 in developmental stages and tissues is closely related to the detoxification of heterologous substances. TcCYP6BQ8 expression was significantly induced after exposure to terpinen-4-ol, and RNA interference against TcCYP6BQ8 increased terpinen-4-ol-induced larval mortality from 47.78 to 66.67%. This indicates that TcCYP6BQ8 may be involved in T. castaneum's metabolism of terpinen-4-ol. Correlation investigation between the CYP6BQ8 gene and terpinen-4-ol resistance in T. castaneum revealed that the TcCYP6BQ8 gene was one of the factors behind T. castaneum's resistance to terpinen-4-ol. This discovery may provide a new theoretical foundation for future regulation of T. castaneum.

Cinobufagin induces acute promyelocytic leukaemia cell apoptosis and PML-RARA degradation in a caspase-dependent manner by inhibiting the ß-catenin signalling pathway.

CONTEXT: Acute promyelocytic leukaemia (APL) is a malignant hematological tumour characterized by the presence of promyelocytic leukaemia-retinoic acid receptor A (PML-RARA) fusion protein. Cinobufagin (CBG) is one of the main effective components of toad venom with antitumor properties. However, only a few reports regarding the CBG treatment of APL are available. OBJECTIVE: We explored the effect and mechanism of action of CBG on NB4 and NB4-R1 cells. MATERIALS AND METHODS: We evaluated the viability of NB4 and NB4-R1 cells treated with 0, 20, 40, and 60 nM CBG for 12, 24, and 48 h. After treatment with CBG for 24 h, Bcl-2 associated X (Bax), B-cell lymphoma 2 (Bcl-2), ß-catenin, cyclin D1, and c-myc expression was detected using western blotting and real-time polymerase chain reaction. Caspase-3 and PML-RARA expression levels were detected using western blotting. RESULTS: CBG inhibited the viability of NB4 and NB4-R1 cells. The IC50 values of NB4 and NB4-R1 cells treated with CBG for 24 h were 45.2 nM and 37.9 nM, respectively. CBG induced NB4 and NB4-R1 cell apoptosis and PML-RARA degradation in a caspase-dependent manner and inhibited the ß-catenin signalling pathway. DISCUSSION AND CONCLUSION: CBG induced NB4 and NB4-R1 cell apoptosis and PML-RARA degradation in a caspase-dependent manner by inhibiting the ß-catenin signalling pathway. This study proposes a novel treatment strategy for patients with APL, particularly those with ATRA-resistant APL.

Functional analysis of a cytochrome P450 gene CYP9Z6 responding to terpinen-4-ol in the red flour beetle, Tribolium castaneum.

Tribolium castaneum is an agricultural and stored pest found throughout the world. The cytochrome P450 genes of T. castaneum can encode various detoxification enzymes and catabolize heterologous substances, conferring tolerance to insecticides. Herein, we describe the identification of a P450 gene (CYP9Z6) from T. castaneum and investigated its expression profile and potential role in the detoxification of terpinen-4-ol. TcCYP9Z6 expression was significantly induced after exposure to terpinen-4-ol, and RNA-mediated silencing of TcCYP9Z6 increased terpinen-4-ol-induced larval mortality from 47.75% to 63.92%, showing that TcCYP9Z6 is closely related to the detoxification of terpinen-4-ol. The developmental expression profile revealed that TcCYP9Z6 was mainly expressed in late adults and late larvae. Tissue expression profiling revealed that the highest TcCYP9Z6 expression occurred in the head, in both the adult and the larval tissues, followed by the gut in larvae and the antennae in adults. These developmental stages and tissues with high TcCYP9Z6 expression are closely related to the detoxification of heterologous substances. These results indicated that TcCYP9Z6 may play a pivotal role in the detoxification of terpinen-4-ol, which provides support for using TcCYP9Z6 a potential gene for the RNAi-mediated prevention and control of T. castaneum.

Aldehyde oxidases mediate plant toxicant susceptibility and fecundity in the red flour beetle, Tribolium castaneum.

Aldehyde oxidases (AOXs) are a group of metabolic enzymes that play critical roles in the degradation of xenobiotics and chemicals. However, the physiological function of this enzyme in insects remains poorly understood. In this study, three TcAOX genes (TcAOX1, TcAOX2, TcAOX3) were identified and characterized from Tribolium castaneum genome. Spatiotemporal expression profiling showed that TcAOX1 expression was most highly expressed at the early pupal stage and was predominantly expressed in the antennae of adults, indicating that TcAOX1 was involved in the degradation of chemical signals; TcAOX2 expression was most highly expressed at the late pupal stage and was mainly expressed in the fat body, epidermis of larvae and adults, respectively; and TcAOX3 expression was in all stages and was primarily expressed in the head of adults. Moreover, the transcripts of TcAOX2 and TcAOX3 were significantly induced after exposure to plant oil, and RNA interference (RNAi) targeting of each of them enhanced the susceptibility of beetles to this plant toxicant, suggesting that these two genes are associated with plant toxicant detoxification. Intriguingly, knockdown of the TcAOX1 led to reductions in female egg-laying but unchanged the hatchability and the development of genital organs, suggesting that this gene may mediate fecundity by effecting the inactivation of chemical signals in T. castaneum. Overall, these results shed new light on the function of AOX genes in insects, and could facilitate the development of research on pest control management.

Altered Functional Connectivity of Hypothalamus in Lifelong Premature Ejaculation Patients.

BACKGROUND: As one of the most prevalent sexual dysfunctions in men, lifelong premature ejaculation (PE) often leads to patient distress. The hypothalamus is implicated in the ejaculatory control of healthy males. However, we do not know whether the hypothalamus-related intrinsic connectivity is altered in lifelong PE patients. PURPOSE: To investigate abnormal intrinsic connectivity of the hypothalamus in lifelong PE patients compared with healthy controls (HCs). STUDY TYPE: Prospective pilot study using cross-sectional data of patients and HCs. SUBJECTS: Forty-seven lifelong PE patients and 30 HCs were included in this study. FIELD STRENGTH/SEQUENCE: 3.0T MRI scanner for T1 -weighted imaging using spoiled gradient recalled echo sequence and functional imaging using a single-shot gradient recalled echo sequence. ASSESSMENT: Preprocessing of MRI data and hypothalamus-seeded functional connectivity (FC) computation were performed using DPABI4.1. STATISTICAL TESTS: The two-sample t-test within SPM12 was adopted to examine possible alterations of intrinsic connectivity of hypothalamus in lifelong PE patients compared with HCs including anxiety and depression scores as covariates (false discovery rate-corrected, P < 0.05). The correlation analysis was then used to assess possible associations between the imaging findings and clinical features in the patient group (Bonferroni-corrected, P < 0.05). RESULTS: Compared with HCs, lifelong PE patients had decreased hypothalamus-seeded FC in the left orbitofrontal cortex, bilateral insula, superior temporal cortex, superior temporal pole, middle temporal cortex, left fusiform, right parahippocampal gyrus, and right cerebellum. The intravaginal ejaculatory latency time positively correlated with the mean z-score from the hypothalamus-insula (r = 0.45) and hypothalamus-cerebellum (r = 0.48) intrinsic connectivity, separately. DATA CONCLUSION: We have shown that hypothalamus-seeded FC alterations and the correlations between the aforementioned abnormal FC alterations and intravaginal ejaculatory latency time. The current findings may promote the understanding of the hypothalamus-related neural mechanisms involved in the abnormal ejaculatory information processing in lifelong PE patients. LEVEL OF EVIDENCE: 1 TECHNICAL EFFICACY STAGE: 3 J. Magn. Reson. Imaging 2020;52:778-784.