The tuber-specific StbHLH93 gene regulates proplastid-to-amyloplast development during stolon swelling in potato.

In potato, stolon swelling is a complex and highly regulated process, and much more work is needed to fully understand the underlying mechanisms. We identified a novel tuber-specific basic helix-loop-helix (bHLH) transcription factor, StbHLH93, based on the high-resolution transcriptome of potato tuber development. StbHLH93 is predominantly expressed in the subapical and perimedullary region of the stolon and developing tubers. Knockdown of StbHLH93 significantly decreased tuber number and size, resulting from suppression of stolon swelling. Furthermore, we found that StbHLH93 directly binds to the plastid protein import system gene TIC56 promoter, activates its expression, and is involved in proplastid-to-amyloplast development during the stolon-to-tuber transition. Knockdown of the target TIC56 gene resulted in similarly problematic amyloplast biogenesis and tuberization. Taken together, StbHLH93 functions in the differentiation of proplastids to regulate stolon swelling. This study highlights the critical role of proplastid-to-amyloplast interconversion during potato tuberization.

Epidemiological characteristics of Epstein-Barr virus infection in children and distribution of disease spectrum in hospitalized patients in shandong province.

This study aims to investigate the epidemiological characteristics of Epstein-Barr virus (EBV) infection in children and the distribution of disease spectrum in hospitalized patients in Shandong Province. A retrospective analysis was conducted on the clinical data of children with EBV infection admitted to hospitals in Shandong Province from January 2022 to December 2022. The epidemiological characteristics, including age, gender, and clinical manifestations, were analyzed. The detection rate of EBV antibodies and the seropositivity rates of different antibodies were also examined. A total of 7124 children with EBV infection were included in this study, with an average age of 7.5 years. The male-to-female ratio was 1.43:1. Among the patients, the positive detection rate of EBV antibodies was 78.40%. The seropositivity rate of Epstein-Barr viral capsid antigen-Immunoglobin G antibodies was 57.09%. The highest incidence of EBV infection was observed in the age group 36-72 months. The urban positive rate was higher than that in rural areas. EBV infection in children in Shandong Province exhibits specific epidemiological characteristics, with a higher incidence in the age group of 36-72 months. Fever, sore throat, and fatigue are the main clinical manifestations. The detection rate of EBV antibodies is relatively high among hospitalized patients. These findings provide valuable information for controlling the transmission of children with suspected Epstein-Barr virus infection.

Injured sensory neurons-derived galectin-3 contributes to neuropathic pain via programming microglia in the spinal dorsal horn.

Emerging studies have demonstrated spinal microglia play a critical role in central sensitization and contribute to chronic pain. Although several mediators that contribute to microglia activation have been identified, the mechanism of microglia activation and its functionally diversified mechanisms in pathological pain are still unclear. Here we report that injured sensory neurons-derived Galectin-3 (Gal3) activates and reprograms microglia in the spinal dorsal horn (SDH) and contributes to neuropathic pain. Firstly, Gal3 is predominantly expressed in the isolectin B4 (IB4)-positive non-peptidergic sensory neurons and significantly up-regulated in dorsal root ganglion (DRG) neurons and primary afferent terminals in SDH in the partial sciatic nerve ligation (pSNL)-induced neuropathic pain model. Gal3 knockout (Gal3 KO) mice showed a significant decrease in mechanical allodynia and Gal3 inhibitor TD-139 produced a significant anti-allodynia effect in the pSNL model. Furthermore, pSNL-induced microgliosis was compromised in Gal3 KO mice. Additionally, intrathecal injection of Gal3 produces remarkable mechanical allodynia by direct activation of microglia, which have enhanced inflammatory responses with TNF-α and IL-1ß up-regulation. Thirdly, using single-nuclear RNA sequencing (snRNA-seq), we identified that Gal3 targets microglia and induces reprogramming of microglia, which may contribute to neuropathic pain establishment. Finally, Gal3 enhances excitatory synaptic transmission in excitatory neurons in the SDH via microglia activation. Our findings reveal that injured sensory neurons-derived Gal3 programs microglia in the SDH and contribute to neuropathic pain.

An Expedient Radical Approach for the Decarboxylative Synthesis of Stereodefined All-Carbon Tetrasubstituted Olefins.

We report a user-friendly approach for the decarboxylative formation of stereodefined and complex tri- and tetra-substituted olefins from vinyl cyclic carbonates and amines as radical precursors. The protocol relies on easy photo-initiated α-amino-radical formation followed by addition onto the double bond of the substrate resulting in a sequence involving carbonate ring-opening, double bond relay, CO2 extrusion and finally O-protonation. The developed protocol is efficient for both mismatched and matched polarity substrate combinations, and the scope of elaborate stereodefined olefins that can be forged including drug-functionalized derivatives is wide, diverse and further extendable to other types of heterocyclic and radical precursors. Mechanistic control reactions show that the decarboxylation step is a key driving force towards product formation, with the initial radical addition under steric control.

Contribution of cuproptosis and Cu metabolism-associated genes to chronic obstructive pulmonary disease.

Airway epithelial cell injury plays a crucial role in the pathogenesis of chronic obstructive pulmonary disease (COPD). However, a novel form of Cu-induced programmed cell death known as cuproptosis has not yet been thoroughly investigated in the context of COPD. Clinical reports have suggested that high copper exposure may increase the risk of COPD. In this study, we aimed to determine the expression and potential functions of cuproptosis-related genes and genes associated with copper metabolism in COPD. We initially identified 52 copper metabolism-related genes based on a review of the literature. Subsequently, we calculated the expression levels of these genes using data from four GEO datasets. To gain insights into the activated signalling pathways and underlying mechanisms in COPD patients, we conducted Gene Ontology (GO) and KEGG pathway analyses, examined protein-protein interactions, and performed weighted correlation network analysis. Our findings revealed that 18 key copper metabolism-related genes, including 5 cuproptosis-related genes, were significantly enriched in signalling pathways and biological processes associated with the development of COPD. Further analysis of clinical data and animal experiments confirmed the high expression of certain cuproptosis key regulators, such as DLD and CDKN2A, in both healthy smokers and COPD smokers. Additionally, these regulators exhibited abnormal expression in a COPD rat model. Notably, copper content was found to be elevated in the lung tissues of COPD rats, suggesting its potential involvement in cuproptosis. These findings provide an experimental foundation for further research into the role of cuproptosis in COPD. Targeting copper metabolism-related genes may represent an effective approach for the treatment of COPD.

PDK1-stabilized LncRNA SPRY4-IT1 promotes breast cancer progression via activating NF-κB signaling pathway.

Pyruvate dehydrogenase kinase 1 (PDK1) is a widely known glycolytic enzyme, and some evidence showed that PDK1 promoted breast cancer by multiple approaches. However, very few lncRNAs have been identified to be associated with PDK1 in breast cancer in previous research. In this study, we found that lncRNA sprouty4-intron transcript 1 (SPRY4-IT1) was regulated by PDK1 with correlation analysis, and PDK1 upregulated SPRY4-IT1 remarkably in breast cancer cells, as PDK1 interacted with SPRY4-IT1 in the nucleus and significantly enhanced the stability of SRPY4-IT1. Furthermore, SPRY4-IT1 was highly expressed in breast cancer, significantly promoted the proliferation and inhibited apoptosis of breast cancer cells. In terms of mechanism, SPRY4-IT1 inhibited the transcription of NFKBIA and the expression of IκBα, thus promoting the formation of p50/p65 complex and activating NF-κB signaling pathway, which facilitated survival of breast cancer cells. Therefore, our finding reveals that PDK1/SPRY4-IT1/NFKBIA axis plays a crucial role that promoting tumor progression, and SPRY4-IT1 knockdown incombined with PDK1 inhibitor is promising to be a new therapeutic strategy in breast cancer.

RGCC balances self-renewal and neuronal differentiation of neural stem cells in the developing mammalian neocortex.

During neocortical development, neural stem cells (NSCs) divide symmetrically to self-renew at the early stage and then divide asymmetrically to generate post-mitotic neurons. The molecular mechanisms regulating the balance between NSC self-renewal and neurogenesis are not fully understood. Using mouse in utero electroporation (IUE) technique and in vitro human NSC differentiation models including cerebral organoids (hCOs), we show here that regulator of cell cycle (RGCC) modulates NSC self-renewal and neuronal differentiation by affecting cell cycle regulation and spindle orientation. RGCC deficiency hampers normal cell cycle process and dysregulates the mitotic spindle, thus driving more cells to divide asymmetrically. These modulations diminish the NSC population and cause NSC pre-differentiation that eventually leads to brain developmental malformation in hCOs. We further show that RGCC might regulate NSC spindle orientation by affecting the organization of centrosome and microtubules. Our results demonstrate that RGCC is essential to maintain the NSC pool during cortical development and suggest that RGCC defects could have etiological roles in human brain malformations.

Focused liquid ultrasonography in dropsy protocol for quantitative assessment of subcutaneous edema.

BACKGROUND: Although subcutaneous edema is a common symptom of critically ill patients, it is still underreported due to the lack of a systematic method for evaluating it. The present study aims to describe the occurrence and distribution of subcutaneous edema, as well as the risk factors associated with it, in critically ill patients using the focused liquid ultrasonography in dropsy (FLUID) protocol, and to assess their impact on ICU mortality. METHODS: The FLUID protocol and the pitting test were performed on general ICU patients in China. Cohen's Kappa coefficient and Bland-Altman plots were used to evaluate the agreement between the two methods at each measurement site and between the whole-body subcutaneous edema scores, respectively, while a repeated measures ANOVA was performed to compare the differences between the two methods in whole-body and body-part measurements. A generalized linear model was used to evaluate the risk factors for subcutaneous edema development and the relationship between subcutaneous edema severity and ICU mortality. RESULTS: A total of 145 critically ill patients were evaluated using both approaches, of whom 40 (27.6%) experienced subcutaneous edema. Over 1440 measurements, it was found that ultrasound discovered more subcutaneous edema than the pitting test (ultrasound: 522[36.3%], pitting test: 444[30.8%], χ2 = 9.477, p = 0.002). The FLUID protocol scored edema severity significantly higher than the pitting test in the whole body and specific body parts, including the abdominal wall, thighs, chest wall, and hands. Subcutaneous edema exhibited gravity-dependent distribution patterns, particularly in the abdominal wall. The APACHE II, NT-proBNP, serum creatinine, and sepsis were independent risk factors for subcutaneous edema development. The score of ultrasonic subcutaneous edema was related to ICU mortality. CONCLUSIONS: The FLUID protocol provides a comprehensive strategy for the semi-quantitative assessment of subcutaneous edema in critically ill patients. In detecting the onset and severity of edema, ultrasound was found to outperform the pitting test. Subcutaneous edema showed a gravity-dependent distribution pattern, and its severity was associated with mortality.

Ugi reaction-assisted assembly of covalent PROTACs against glutathione peroxidase 4.

Inducing cell ferroptosis by inactivating glutathione peroxidase 4 (GPX4) is a popular cancer treatment strategy. However, only few GPX4 inhibitors have been developed to date. PROteolysis Targeting Chimera (PROTAC) is a promising approach to provide new opportunities to overcome limitations of traditional therapeutics. Herein, a PROTAC-like activity-based probe PD-Q2 was first assembled using Ugi reaction, consisting of a known GPX4 inhibitor ML-162 homolog to the E3 ligase cereblon ligand-pomalidomide. Pull-down and immunoblotting analysis revealed that GPX4 was a covalent target of PD-Q2, but the degradation efficiency was weak. Therefore, a series of degraders was further synthesized by varying the linkers of heterofunctional PROTACs. Among these degraders, PD-4 and PD-P2 were found to promote effective GPX4 degradation via the ubiquitin-proteasome system and cause lipid ROS accumulation. PD-4 and PD-P2 showed potent inhibitory of colony formation and cell growth. Furthermore, we found that with pomalidomide, the degraders exhibit a high fluorescent signal that is mostly localized in the lysosome, which may affect the effectiveness of anti-cell proliferation. Overall, we provide GPX4 degraders for further exploring therapeutic potential of regulating ferroptosis.

Dual-aptamer-engineered M1 macrophage with enhanced specific targeting and checkpoint blocking for solid-tumor immunotherapy.

Chimeric antigen receptor T (CAR-T) cell therapy has faced a series of challenges and has shown very little efficacy in solid tumors to date. Although genetically engineered macrophages have achieved definite therapeutic effect in solid tumors, heterogeneous expression of engineered proteins and the potential for toxicity limit further applications. Herein, we propose a nongenetic and simple macrophage cell engineering strategy through glycan metabolic labeling and click reaction for the treatment of solid tumors. The aptamer-engineered M1 macrophage (ApEn-M1) showed enhanced active targeting ability for tumor cells in vitro and in vivo, resulting in significant cytotoxicity effects. Moreover, ApEn-M1 exhibited superior antitumor efficacy in a breast cancer xenograft mouse model and a lung metastasis mouse model of breast cancer. Interestingly, the ApEn-M1 could reprogram the immunity microenvironment by increasing T cell infiltration and enhancing T cell activity in the tumor region. Additionally, the administration of ApEn-M1 showed no obvious systemic side effects. With glycan metabolic labeling, the macrophages could be efficiently labeled with aptamers on the cell surface via click reaction without genetic alteration or cell damage. Hence, this study serves as a proof of concept for cell-surface anchor engineering and expands the range of nongenetic macrophage cell engineering strategies.

Trunk rot of Juglans regia caused by Myrmaecium fulvopruinatum in China.

Walnut (Juglans regia) is a deciduous tree of the Juglandaceae family, widely cultivated in China, and provides value in a variety of ways, including the usage of the wood and nuts, and offers substantial economic, social, and environmental advantages (Wang et al, 2017). Nevertheless, a fungal disease of causing walnut trunk rot was observed in approximately 30% of 50 counted ten-year-old J. regia in Chongzhou City (30°33'34″N, 103°38'35″E, 513 m), Sichuan Province, China, and this disease has greatly delete healthy growth of walnut. The infected bark exhibited purple necrotic lesions, and the sick parts were surrounded by water-soaked plaques. From 10 trunks of the 10 diseased trees, 20 isolated fungal colonies were the same. The ascospores placed in 60 mm plates were almost entirely covered with mycelium within 8 days, colonies on the PDA changed from initial pale to white, ad then turned yellowish to light orange or rosy to yellow-brown (25℃, 90% relative humidity, 12-h photoperiod). On the host, Ectostromata were immersed to erumpent, globose to subglobose, purple and brown, and measured 0.6 - 4.5 × 0.3 - 2.8 mm (x̄ = 2.6 × 1.6 mm, n = 40); Ascomata were flask-shaped to subglobose, dark brown, and measured 0.1 - 0.6 × 0.1 - 0.4 mm (x̄ = 0.35 × 0.25 mm, n = 40); Asci were numerous, cylindrical to subclavate, contained 8 uniseriate ascospores, and measured 80 - 150 × 10 - 20 µm (x̄ = 115 × 15 µm, n = 40), and Ascospores were ellipsoid, 2-celled, dark brown to black, plump or attenuated towards, apices with 1 large drop per cell, and measured 14 - 20 × 6.5 - 9 µm (x̄ = 17 × 7.8 µm, n = 40). These morphological characteristics are consistent with the species Myrmaecium fulvopruinatum (Berk.) Jaklitsch & Voglmayr (Jaklitsch et al. 2015). The genomic DNA of a representative isolate SICAUCC 22-0148 was extracted. The ITS, LSU region, tef1-α, rpb2 genes region were amplified using the primer pairs ITS1/ITS4 primers (White et al. 1990), LR0R/LR5 (Moncalvo et al. 1995), EF1-688F/986R (Alves et al. 2008), fRPB2-5f/fRPB2-7cr (Liu et al. 1999), respectively. The sequences were deposited in NCBI with accession numbers ON287043 (ITS), ON287044 (LSU), ON315870 (tef1-α), and ON315871 (rpb2), rspectively, which showed 99.8, 99.8, 98.1, and 98.5% identities with M. fulvopruinatum CBS 139057 holotype (accession numbers KP687858, KP687858, KP688027, and KP687933 respectively). Based on the analyses of phylogenies and morphologies, the isolates were identified as M. fulvopruinatum. The pathogenicity of SICAUCC 22-0148 was tested by inoculating surface-sterilized trunk wounds of four-year-old trees of J. regia with a mycelial plug (Desai et al. 2019). Sterile PDA plugs were used as controls. Wounds were covered with a film, to ensure humidity and prevent contamination. Each inoculation was repeated twice and included two plants, control and inoculated. A month later, the symptoms observed on inoculated trunks were similar to those in the wild, and M. fulvopruinatum was re-isolated from the inoculated trunk, confirming Koch's postulates. Previous research has reported M. fulvopruinatum as an important fungal species that cause canker delete symptoms on Chinese sweet chestnut in China (Jiang et al. 2018). We carried the taxonomy work of the fungi that caused trunk rot on walnut, and this is the first time that M. fulvopruinatum has been linked to walnut trunk rot on J. regia. Trunk rot of walnut will not only cause weakening of trees, but also affect the yield and quality of walnuts, bringing huge economic losses. This study was supported by the Sichuan Science and Technology Program under Grant 2022NSFSC1011. References: Alves, A., et al. 2008. Fungal Diversity 28:1-13. Desai, D.D., et al. 2019. International Journal of Economic Plants 6:147-149. Jaklitsch., W.M., et al. 2015. Fungal Diversity 73(1):159-202. Jiang, N., et al. 2018. Mycosphere 9(6):1268-1289. Liu, Y.L., et al. 1999. Mol Biol Evol 16:1799-1808. Moncalvo, J.M., et al. 1995. Mycologia 87:223-238. Wang, Q.H., et al. 2017. Australasian Plant Pathology 46:585-595. White, T.J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA.

Meta-analysis of the efficacy and safety of sevelamer as hyperphosphatemia therapy for hemodialysis patients.

This study was designed to examine the relative safety and efficacy of sevelamer in the treatment of chronic kidney disease (CKD) patients in comparison to placebo, calcium carbonate (CC), or lanthanum carbonate (LC). The PubMed, Embase, Cochrane Library, and China National Knowledge Infrastructure (CNKI) databases were searched for articles published through 18 June 2022. The quality of relevant studies was independently analyzed by two investigators who also extracted data from these manuscripts as per Cochrane Collaboration Handbook 5.3. The safety and efficacy of sevelamer as a treatment for hyperphosphatemia in CKD patients were then examined through a meta-analysis, with the primary patient-level outcomes of interest in this analysis being all-cause mortality and the incidence of gastrointestinal adverse effects. Vascular calcification score was also examined as an intermediate outcome, while serum biochemical parameters including levels of phosphate (P), calcium (Ca), intact parathyroid hormone (iPTH), lipids, C-reactive protein (CRP), or fibroblast growth factor-23 (FGF-23) were additionally assessed. In total, this meta-analysis incorporated data from 34 randomized controlled trials (RCTs) enrolling 2802 patients. Sevelamer was associated with reduced all-cause mortality (RR 0.28, CI 0.19 - 0.41, very low certainty) and Vessel calcification score (RR -0.58, CI -1.11 to -0.04, low certainty) and induced less hypercalcemia (MD -0.28, CI 0.40 to -0.16, low certainty) and hyperphosphatemia (MD -0.22, CI -0.32 to -0.13, low certainty) when compared with Ca-based binders in CKD5D individuals. No significant differences in gastrointestinal adverse events (GAEs) incidence were observed. These data suggest that sevelamer may represent a beneficial means of protecting CKD patients against death and vessel calcification when used to treat hyperphosphatemia, while we found no clinically important benefits in decreasing gastrointestinal adverse effects.

Fast CU Partition Algorithm for Intra Frame Coding Based on Joint Texture Classification and CNN.

High-efficiency video coding (HEVC/H.265) is one of the most widely used video coding standards. HEVC introduces a quad-tree coding unit (CU) partition structure to improve video compression efficiency. The determination of the optimal CU partition is achieved through the brute-force search rate-distortion optimization method, which may result in high encoding complexity and hardware implementation challenges. To address this problem, this paper proposes a method that combines convolutional neural networks (CNN) with joint texture recognition to reduce encoding complexity. First, a classification decision method based on the global and local texture features of the CU is proposed, efficiently dividing the CU into smooth and complex texture regions. Second, for the CUs in smooth texture regions, the partition is determined by terminating early. For the CUs in complex texture regions, a proposed CNN is used for predictive partitioning, thus avoiding the traditional recursive approach. Finally, combined with texture classification, the proposed CNN achieves a good balance between the coding complexity and the coding performance. The experimental results demonstrate that the proposed algorithm reduces computational complexity by 61.23%, while only increasing BD-BR by 1.86% and decreasing BD-PSNR by just 0.09 dB.

Concentrated growth factor combined with iRoot BP Plus promotes inflamed pulp repair: an in vitro and in vivo study.

BACKGROUND: Platelet concentrates combined with calcium silicate cements may promote reparative dentin formation. However, few studies have reported their effect on dental pulp inflammation. This study aimed to evaluate the effects of concentrated growth factor (CGF) combined with iRoot BP Plus on inflammatory human dental pulp stem cells (hDPSCs) in vitro and inflamed pulp in rats in vivo. METHODS: The proliferation of LPS-stimulated hDPSCs treated with 50% CGF with/without 25% iRoot BP Plus was evaluated using Cell Counting Kit-8 on days 1, 4 and 7. The expression of genes associated with inflammation on day 1 and differentiation on day 14 was analysed by real-time polymerase chain reaction. The exposed pulp of rat maxillary molars was injected with 10 mg/mL LPS and directly capped with CGF membrane with/without iRoot BP Plus extract for 1, 7 and 28 days. The teeth were subjected to histologic analyses and immunohistochemistry. RESULTS: The proliferation rates of the inflammatory hDPSCs after the combination treatment were significantly higher than those after the other treatments on days 4 and 7 (P < 0.05). IL-1ß, IL-6, and TNF-α levels were increased in inflammatory hDPSCs but decreased after treatment with CGF combined with iRoot BP Plus extract, whereas IL-4 and IL-10 showed the opposite expression patterns. Expression of the odontogenesis-related genes OCN, Runx2, and ALP was dramatically enhanced by combined treatment with CGF and iRoot BP Plus extract. In rat pulp, the average inflammation scores of the CGF and CGF-iRoot BP Plus groups significantly decreased in comparison with those of the LPS group (P < 0.05), and the CGF-iRoot BP Plus group had more reparative dentin than the CGF and BP groups. Immunohistochemical staining showed fewer M1 macrophages on day 1 and more M2 macrophages on day 7 in the CGF-iRoot BP Plus group than in the other groups. CONCLUSIONS: The combination of CGF and iRoot BP Plus showed a synergistic effect on anti-inflammatory potential and promoted greater pulp healing than CGF or iRoot BP Plus alone.

Improvement of Error Correction in Nonequilibrium Information Dynamics.

Errors are inevitable in information processing and transfer. While error correction is widely studied in engineering, the underlying physics is not fully understood. Due to the complexity and energy exchange involved, information transmission should be considered as a nonequilibrium process. In this study, we investigate the effects of nonequilibrium dynamics on error correction using a memoryless channel model. Our findings suggest that error correction improves as nonequilibrium increases, and the thermodynamic cost can be utilized to improve the correction quality. Our results inspire new approaches to error correction that incorporate nonequilibrium dynamics and thermodynamics, and highlight the importance of the nonequilibrium effects in error correction design, particularly in biological systems.

Nonequilibrium Effects on Information Recoverability of the Noisy Channels.

We investigated the impact of nonequilibrium conditions on the transmission and recovery of information through noisy channels. By measuring the recoverability of messages from an information source, we demonstrate that the ability to recover information is connected to the nonequilibrium behavior of the information flow, particularly in terms of sequential information transfer. We discovered that the mathematical equivalence of information recoverability and entropy production characterizes the dissipative nature of information transfer. Our findings show that both entropy production (or recoverability) and mutual information increase monotonically with the nonequilibrium strength of information dynamics. These results suggest that the nonequilibrium dissipation cost can enhance the recoverability of noise messages and improve the quality of information transfer. Finally, we propose a simple model to test our conclusions and found that the numerical results support our findings.

The miR-345-3p/PPP2CA signaling axis promotes proliferation and invasion of breast cancer cells.

Breast cancer is the most common malignancy among women worldwide. Functional studies have demonstrated that miRNA dysregulation in many cases of cancer, in which miRNAs act as either oncogenes or tumor suppressor. Here we report that miR-345-3p is generally upregulated in breast cancer tissues and breast cancer cell lines. Overexpression and inhibition of miR-345-3p revealed its capacity in regulating proliferation and invasion of breast cancer cells. Further research identified protein phosphatase 2 catalytic subunit alpha (PPP2CA), a suppressor of AKT phosphorylation, as a candidate target of miR-345-3p. In vitro, miR-345-3p mimics promoted AKT phosphorylation by targeting its negative regulator, PPP2CA. Blocking miR-345-3p relieved its inhibition of PPP2CA, which attenuated PI3K-AKT signaling pathway. In vivo, inhibiting miR-345-3p by miR-345-3p-inhibition lentivirus suppressed tumor growth and invasiveness in mice. Together, the miR-345-3p/PPP2CA signaling axis exhibits tumor-promoting functions by regulating proliferation and invasion of breast cancer cells. These data provide a clue to novel therapeutic approaches for breast cancer.

Epithelia-Sensory Neuron Cross Talk Underlies Cholestatic Itch Induced by Lysophosphatidylcholine.

BACKGROUND & AIMS: Limited understanding of pruritus mechanisms in cholestatic liver diseases hinders development of antipruritic treatments. Previous studies implicated lysophosphatidic acid (LPA) as a potential mediator of cholestatic pruritus. METHODS: Pruritogenicity of lysophosphatidylcholine (LPC), LPA's precursor, was examined in naïve mice, cholestatic mice, and nonhuman primates. LPC's pruritogenicity involving keratinocyte TRPV4 was studied using genetic and pharmacologic approaches, cultured keratinocytes, ion channel physiology, and structural computational modeling. Activation of pruriceptor sensory neurons by microRNA-146a (miR-146a), secreted from keratinocytes, was identified by in vitro and ex vivo Ca2+ imaging assays. Sera from patients with primary biliary cholangitis were used for measuring the levels of LPC and miR-146a. RESULTS: LPC was robustly pruritic in mice. TRPV4 in skin keratinocytes was essential for LPC-induced itch and itch in mice with cholestasis. Three-dimensional structural modeling, site-directed mutagenesis, and channel function analysis suggested a TRPV4 C-terminal motif for LPC binding and channel activation. In keratinocytes, TRPV4 activation by LPC induced extracellular release of miR-146a, which activated TRPV1+ sensory neurons to cause itch. LPC and miR-146a levels were both elevated in sera of patients with primary biliary cholangitis with itch and correlated with itch intensity. Moreover, LPC and miR-146a were also increased in sera of cholestatic mice and elicited itch in nonhuman primates. CONCLUSIONS: We identified LPC as a novel cholestatic pruritogen that induces itch through epithelia-sensory neuron cross talk, whereby it directly activates skin keratinocyte TRPV4, which rapidly releases miR-146a to activate skin-innervating TRPV1+ pruriceptor sensory neurons. Our findings support the new concept of the skin, as a sensory organ, playing a critical role in cholestatic itch, beyond liver, peripheral sensory neurons, and central neural pathways supporting pruriception.

Osimertinib is an effective epidermal growth factor receptor-tyrosine kinase inhibitor choice for lung cancer with epidermal growth factor receptor exon 18-25 kinase domain duplication: report of two cases.

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are an effective treatment for common EGFR mutations in non-small-cell lung cancer (NSCLC). Rarer EGFR mutations such as kinase domain duplications (KDDs) have been identified, but the optimal therapy following treatment resistance remains unknown. We report two patients who were diagnosed with NSCLC including KDD. For case 1, afatinib (40 mg once daily) was at first effective but then became ineffective. Consequently, osimertinib therapy (80 mg once daily) was administered. As of 26 May 2021, the osimertinib therapy achieved a stable disease state according to the chest computed tomography scan. As for case 2, the patient received second-line chemotherapy and anlotinib (12 mg once daily) for 6 months and died in May 2020. Here, we describe osimertinib as an effective therapy for EGFR-KDD positive lung adenocarcinoma and thereby provide a new alternative for further treatment following resistance to first- and second-generation EGFR-TKIs.

Lupeol protects against cardiac hypertrophy via TLR4-PI3K-Akt-NF-κB pathways.

Inflammation and apoptosis are main pathological processes that lead to the development of cardiac hypertrophy. Lupeol, a natural triterpenoid, has shown anti-inflammatory and anti-apoptotic activities as well as potential protective effects on cardiovascular diseases. In this study we investigated whether lupeol attenuated cardiac hypertrophy and fibrosis induced by pressure overload in vivo and in vitro, and explored the underlying mechanisms. Cardiac hypertrophy was induced in mice by transverse aortic constriction (TAC) surgery, and in neonatal rat cardiomyocytes (NRCMs) by stimulation with phenylephrine (PE) in vitro. We showed that administration of lupeol (50 mg ·kg-1· d-1, i.g., for 4 weeks) prevented the morphological changes and cardiac dysfunction and remodeling in TAC mice, and treatment with lupeol (50 µg/mL) significantly attenuated the hypertrophy of PE-stimulated NRCMs, and blunted the upregulated hypertrophic markers ANP, BNP, and ß-MHC. Furthermore, lupeol treatment attenuated the apoptotic and inflammatory responses in the heart tissue. We revealed that lupeol attenuated the inflammatory responses including the reduction of inflammatory cytokines and inhibition of NF-κB p65 nuclear translocation, which was mediated by the TLR4-PI3K-Akt signaling. Administration of a PI3K/Akt agonist 740 Y-P reversed the protective effects of lupeol in TAC mice as well as in PE-stimulated NRCMs. Moreover, pre-treatment with a TLR4 agonist RS 09 abolished the protective effects of lupeol and restored the inhibition of PI3K-Akt-NF-κB signaling by lupeol in PE-stimulated NRCMs. Collectively, our results demonstrate that the lupeol protects against cardiac hypertrophy via anti-inflammatory mechanisms, which results from inhibiting the TLR4-PI3K-Akt-NF-κB signaling.