[Effects of 4-hydroxy-2(3H)-benzoxazolone on proliferation and apoptosis of pancreatic cancer L3.6 cells].

This study explored the effects of 4-hydroxy-2(3H)-benzoxazolone(HBOA) on the proliferation and apoptosis of pancreatic cancer cells and its molecular mechanism. The L3.6 cells cultured in vitro were treated with HBOA of 0-1.0 mmol·L~(-1). The cell viability was detected by the cell counting kit-8(CCK-8) method, and the half inhibitory concentration(IC_(50)) was analyzed to determine the drug concentration and time. The cell morphology was observed under an inverted microscope and by acridine orange(AO) staining. The ability of proliferation and self-renewal were evaluated through live cell counting and colony formation experiments. The cell cycle progression and cell apoptosis rate were detected by flow cytometry. The morphology of cell apoptosis was observed by scanning electron microscopy. The mRNA expression of proliferating cell nuclear antigen(PCNA), cyclinA1, cyclinA2, cyclin dependent kinase 2(CDK2), and cyclin dependent kinase inhibitor 1A(P21) were determined by qPCR. The level of reactive oxygen species(ROS), lipid peroxide, and mitochondrial membrane potential were measured by flow cytometry. The activity of protein kinase B(Akt)/mammalian target of rapamycin(mTOR) signaling pathway was detected by Western blot. Compared with the control group, the cells treated with HBOA exhibited a significant decrease in viability. Then the optimal concentration and intervention time of HBOA were determined to be 0.4 mmol·L~(-1), 0.6 mmol·L~(-1), and 48 h. Compared with the control group, groups with HBOA of 0.4 mmol·L~(-1 )and 0.6 mmol·L~(-1) showed a significant suppression in cell proliferation and colony formation ability, down-regulated mRNA of PCNA, cyclinA1, cyclinA2, and CDK2, up-regulated P21 mRNA, S-phase cell cycle arrest, and increased cell apoptosis rate. There was an appearance of apoptotic bodies, increased ROS and lipid peroxide, decreased mitochondrial membrane potential(with a significant decrease in 0.6 mmol·L~(-1) group), and down-regulated p-Akt and p-mTOR proteins. The results show that HBOA inhibits the proliferation of pancreatic cancer L3.6 cells and induces cell apoptosis, which may be related to the increase in reactive oxygen species and the inhibition of the Akt/mTOR pathway.

Identification and clinical value of a new ceRNA axis (TIMP3/hsa-miR-181b-5p/PAX8-AS1) in thyroid cancer.

Background: Thyroid cancer (TC) is a prevalent and increasingly common malignant tumor. In most cases, TC progresses slowly and runs a virtually benign course. However, challenges remain with the treatment of refractory TC, which does not respond to traditional management or is subject to relapse or metastasis. Therefore, new therapeutic regimens for TC patients with poor outcomes are urgently needed. Methods: The differentially expressed RNAs were identified from the expression profile data of RNA from TC downloaded from The Cancer Genome Atlas database. Multiple databases were utilized to investigate the regulatory relationship among RNAs. Subsequently, a competitive endogenous RNA (ceRNA) network was established to elucidate the ceRNA axis that is responsible for the clinical prognosis of TC. To understand the potential mechanism of ceRNA axis in TC, location analysis, functional enrichment analysis, and immune-related analysis were conducted. Results: A ceRNA network of TC was constructed, and the TIMP3/hsa-miR-181b-5p/PAX8-AS1 ceRNA axis associated with the prognosis of TC was successfully identified. Our results showed that the axis might influence the prognosis of TC through its regulation of regulating tumor immunity. Conclusions: Our findings provide evidence that TIMP3/hsa-miR-181b-5p/PAX8-AS1 axis is significantly related to the prognosis of TC. The molecules involved in this axis may serve as novel therapeutic approaches for TC treatment.

A sensitive electrochemical sensor for chiral detection of tryptophan enantiomers by using carbon black and ß­cyclodextrin.

A chiral sensor for the electrochemical identification of tryptophan (Trp) isomers is described. The electrochemical sensor was prepared based on the combination of (a) carbon black (CB-COOH) as conductive material, (b) Cu2+-modified ß-cyclodextrin (Cu-ß-CD), and (c) ß-CD-based metal-organic frameworks (ß-CD-MOF) as chiral selectors. The Cu-ß-CD can be self-assembled into the CB-COOH and ß-CD-MOF through electrostatic interactions, which was characterized by zeta potential analysis. UV-vis spectroscopy proved that Cu-ß-CD displays a higher combination for D-Trp than L-Trp, and the ß-CD-MOF at the surface of the GCE has a higher affinity for L-Trp than D-Trp, which endow an easier permeation of L-Trp to the surface of the electrode, thus leading to a larger electrochemical signal of differential pulse voltammetry (DPV). The enantioselectivity for L-Trp over D-Trp (IL/ID) is 2.13, with a low detection limit for D-Trp (11.18 µM) and L-Trp (5.48 µM). In addition, the proposed chiral sensor can be chosen to determine  the percentage of D-Trp in enantiomer mixture solutions and real sample detection with a recovery from 98.2 to 102.8% for L-Trp and 97.9 to 101.1% for D-Trp.

Conversion of senescent cartilage into a pro-chondrogenic microenvironment with antibody-functionalized copper sulfate nanoparticles for efficient osteoarthritis therapy.

The development of osteoarthritis (OA) correlates with the expansion of senescent cells in cartilage, which contributes to an inflammatory microenvironment that accelerates matrix degradation and hampers cartilage generation. To address OA, we synthesized small copper sulfide nanoparticles functionalized with anti-beta-2-microglobulin antibodies (B2M-CuS NPs) that catalyze the formation of toxic •OH from H2O2 via peroxidase-like activity. These B2M-CuS NPs are specifically targeted to induce apoptosis in senescent chondrocytes while showing no toxicity toward normal chondrocytes. Furthermore, B2M-CuS NPs enhance the chondrogenesis of normal chondrocytes. Thus, B2M-CuS NPs can effectively treat OA by clearing senescent chondrocytes and promoting cartilage regeneration after intra-articular injection into the knee joints of surgery-induced OA mice. This study uses smart nanomaterials to treat OA with a synergistic strategy that both remodels senescent cartilage and creates a pro-chondrogenic microenvironment.

Improvement of Luminescence Properties of Eulytite Single-Phase White Emitting Ca3Bi (PO4)3: Ce3+/Dy3+ Phosphor.

To reduce the issue of tri-primary color reabsorption, a new approach for single-phase phosphors as light-emitting diodes (LEDs) has been recommended. The structures, morphology, photoluminescence, thermal stability, and luminescence mechanism of a variety of Ca3Bi (PO4)3 (CBPO): Ce3+/Dy3+ phosphors were investigated. XRD characterization showed that all CBPO samples were eulytite structures. Furthermore, the energy transfer process from Ce3+ to Dy3+ in CBPO is systematically investigated in this work, and the color of light can be adjusted by changing the ratio of doped ions. Under UV light, energy is transferred from Ce3+-Dy3+ mainly through quadrupole-quadrupole interactions in the CBPO host, and doping with different Dy3+ concentrations tunes the emission color from blue to white. The thermal stability of the CBPO: 0.04Ce3+, 0.08Dy3+ samples is outstanding, and the CIE coordinates of the samples after emission have little effect with temperature, while their emission intensity at 423 K is as strong as that at room temperature, reaching 90%. The above results indicate that this CBPO material has great potential as a white light phosphor under near-UV excitation at the optimized concentration of Ce3+ and Dy3+.

Pluma, a new genus of slug moths (Lepidoptera: Limacodidae) from South China, with descriptions of two new species.

The genus Pluma, gen. nov. is established to accommodate two new species of limacodid moths, P. shuni sp. nov. and P. yuensis sp. nov., from South China. Based on morphological and molecular characters, the species cannot be placed in any existing genus and therefore they placed in the newly erected one. The new taxa are supported by morphological characters and DNA barcode data. Male adults, including wing venation and genitalia, are illustrated, along with a barcode-based tree.

Inhibition of NOX4 with GLX351322 alleviates acute ocular hypertension-induced retinal inflammation and injury by suppressing ROS mediated redox-sensitive factors activation.

Reactive oxygen species (ROS) overproduction plays an essential role in the etiology of ischemic/hypoxic retinopathy caused by acute glaucoma. NADPH oxidase (NOX) 4 was discovered as one of the main sources of ROS in glaucoma. However, the role and potential mechanisms of NOX4 in acute glaucoma have not been fully elucidated. Therefore, the current study aims to investigate the NOX4 inhibitor GLX351322 that targets NOX4 inhibition in acute ocular hypertension (AOH)-induced retinal ischemia/hypoxia injury in mice. Herein, NOX4 was highly expressed in AOH retinas, particularly the retinal ganglion cell layer (GCL). Importantly, the NOX4 inhibitor GLX351322 reduced ROS overproduction, inhibited inflammatory factor release, suppressed glial cell activation and hyperplasia, inhibited leukocyte infiltration, reduced retinal cell senescence and apoptosis in damaged areas, reduced retinal degeneration and improved retinal function. This neuroprotective effect is at least partially associated with mediated redox-sensitive factor (HIF-1α, NF-κB, and MAPKs) pathways by NOX4-derived ROS overproduction. These results suggest that inhibition of NOX4 with GLX351322 attenuated AOH-induced retinal inflammation, cellular senescence, and apoptosis by inhibiting the activation of the redox-sensitive factor pathway mediated by ROS overproduction, thereby protecting retinal structure and function. Targeted inhibition of NOX4 is expected to be a new idea in the treatment of acute glaucoma.

Ethyl caffeate inhibits macrophage polarization via SIRT1/NF-κB to attenuate traumatic heterotopic ossification in mice.

Heterotopic ossification (HO) denotes the presence of mature bone tissue in soft tissues or around joints. Inflammation is a key driver of traumatic HO, and macrophages play an important role in this process. Ethyl caffeate (ECF), a critical active compound found in Petunia, exerts significant anti-inflammatory effects. Herein, we established a mouse model of HO by transection of the Achilles tendon and back burn and found abundant macrophage infiltration in the early stage of HO, which decreased with time. In vitro and in vivo experiments indicated that ECF inhibited macrophage polarization, and mechanistic studies showed that it inhibited the SIRT1/NF-κB signalling pathway, thereby suppressing the release of downstream inflammatory cytokines. ECF reduced HO in mice, and its effect was comparable to indomethacin (INDO). In vitro studies revealed that ECF did not directly affect the mineralization of mesenchymal stem cells (MSCs) or osteogenic differentiation but inhibited these processes by reducing the level of inflammatory cytokines in the conditioned medium (CM). Thus, M1 macrophages may play a crucial role in the pathogenesis of HO, and ECF is a prospective candidate for the prevention of trauma-induced HO. DATA AVAILABILITY: Data will be made available on request.

Casticin suppresses RANKL­induced osteoclastogenesis and prevents ovariectomy­induced bone loss by regulating the AKT/ERK and NF­κB signaling pathways.

Postmenopausal osteoporosis is a systemic metabolic disease that chronically endangers public health and is typically characterized by low bone mineral density and marked bone fragility. The excessive bone resorption activity of osteoclasts is a major factor in the pathogenesis of osteoporosis; therefore, strategies aimed at inhibiting osteoclast activity may prevent bone decline and attenuate the process of osteoporosis. Casticin (Cas), a natural compound, has anti­inflammatory and antitumor properties. However, the role of Cas in bone metabolism remains largely unclear. The present study found that the receptor activator of nuclear factor­κΒ (NF­κB) ligand­induced osteoclast activation and differentiation were inhibited by Cas. Tartrate­resistant acid phosphatase staining revealed that Cas inhibited osteoclast differentiation, and bone resorption pit assays demonstrated that Cas affected the function of osteoclasts. Cas significantly reduced the expression of osteoclast­specific genes and related proteins, such as nuclear factor of activated T cells, cytoplasmic 1 and c­Fos at the mRNA and protein level in a concentration­dependent manner. Cas inhibited osteoclast formation by blocking the AKT/ERK and NF­κB signaling pathways, according to the intracellular signaling analysis. The microcomputed tomography and tissue staining of tibiae from ovariectomized mice revealed that Cas prevented the bone loss induced by estrogen deficiency and reduced osteoclast activity in vivo. Collectively, these findings indicated that Cas may be used to prevent osteoporosis.

Understanding the cadmium passivation and nitrogen mineralization of aminated lignin in soil.

Aminated lignin (AL) was prepared and first applied to remediation of cadmium (Cd) pollution in soil. Meanwhile, the nitrogen mineralization characteristics of AL in soil and its effect on soil physicochemical properties were elucidated by soil incubation experiment. The results showed that the Cd availability was dramatically lowered in soil by adding the AL. The DTPA-extractable Cd content of AL treatments was considerably reduced by 40.7-71.4 %. The soil pH (5.77-7.01) and absolute value of zeta potential (30.7-34.7 mV) enhanced simultaneously as the AL additions increased. The content of soil organic matter (SOM) (99.0-264.0 %) and total nitrogen (95.9-301.3 %) were gradually enhanced due to high C (63.31 %) and N (9.69 %) content in AL. Moreover, AL significantly elevated the content of mineral nitrogen (77.2-142.4 %) and available nitrogen (95.5-301.7 %). The first-order kinetic equation of soil nitrogen mineralization revealed that AL greatly enhanced nitrogen mineralization potential (84.7-143.9 %) and reduced environmental pollution by lowering the loss of soil inorganic nitrogen. AL could effectively reduce the availability of Cd through direct (self-adsorption) and indirect effects (improvement of soil pH, SOM and reduction of soil zeta potential), thereby achieving passivation of Cd in soil. In short, this work will develop a novel approach and technical support for soil heavy metal remediation, which is of great significance for improving the sustainable development of agricultural production.

UPLC/Q-TOF-MS-based metabolomics evaluate the efficacy of oroxylin A against postmenopausal osteoporosis.

Post-menopausal osteoporosis (PMOP) is a common metabolic bone malady characterized by bone mass loss and bone microarchitectural deterioration; however, there is currently no effective drug for its management. According to our previous study, oroxylin A (OA) could effectively protect ovariectomized (OVX)-osteoporotic mice from bone loss; however, its therapeutic targets are still unclear. From a metabolomic perspective, we studied serum metabolic profiles to discover potential biomarkers and OVX-related metabolic networks, which could assist us to comprehend the impact of OA on OVX. Five metabolites were identified as biomarkers associated with 10 related metabolic pathways, including phenylalanine, tyrosine and tryptophan biosynthesis, and phenylalanine, tryptophan and glycerophospholipid metabolism. After OA treatment, the expression of multiple biomarkers changed, with lysophosphatidylcholine (18:2) being a major significantly regulated biomarker. Our study demonstrated that OA's effects on OVX are probably related to the regulation of phenylalanine, tyrosine and tryptophan biosynthesis. Our findings explain the role of OA against PMOP in terms of metabolism and pharmacology and provide a pharmacological foundation for OA treatment of PMOP.

Corylifol A protects against ovariectomized-induced bone loss and attenuates RANKL-induced osteoclastogenesis via ROS reduction, ERK inhibition, and NFATc1 activation.

Osteoclast differentiation and function are critical targets for anti-osteoporosis treatment. Oxidative stress also plays an important regulatory role in the differentiation of osteoclasts. Corylifol A (CA) is a flavonoid extracted from the Psoralea fruit. It has anti-inflammatory and antioxidant properties despite its unknown effect on osteoporosis. This study found that CA prevented estrogen-deficiency-induced bone loss and suppressed osteoclastogenesis in ovariectomized (OVX) mice by inhibiting intracellular reactive oxygen species (ROS) levels. In vivo, CA effectively prevented trabecular bone loss and reduced osteoclasts' number on the bone surface in OVX mice, as demonstrated in micro-CT, osteometry, and immunohistochemical data. However, CA did not affect cortical bone. In vitro, CA inhibited RANKL-induced podosome belt formation, osteoclastogenesis, and bone resorption functions. CA suppressed RANKL-induced ROS by boosting antioxidant enzymes (Catalase and NQO1) and NFATc1 signaling pathway related protein expression, including integrin αvß3, NFATc1 and CTSK. Moreover, CA inhibited osteoclast-specific genes, including Ctsk, Acp5, and Mmp9. CA also attenuated the MAPK/ERK pathway, but did not affect the NF-κB signaling pathway. In terms of osteogenesis, CA did not inhibit or promote osteogenic differentiation and mineralization in vitro. These results reveal that CA could be a new replacement therapy for treating estrogen-deficiency osteoporosis via suppressing osteoclastogenesis and intracellular ROS.

Primary pulmonary hyalinizing clear cell carcinoma with pseudopapillary structures and abundant cysts filled with mucus.

Pulmonary hyalinizing clear cell carcinoma (HCCC) is a new and rare form of lung salivary gland tumor. Only twenty-two cases have been reported in the literature to date. Furthermore, their clinicopathological features have not been fully characterized. In this paper, we describe the clinicopathological characteristics, immunohistochemical features, and molecular genetic changes in two HCCC cases. We also simultaneously reviewed related literature on similar cases reported. Of the two cases, one was of a 58-year-old man with a 4.3 cm lung tumor, which was the largest among all previously reported cases. The tumor showed an infiltrative growth pattern and perineural and vascular invasion microscopically. Moreover, nuclear grooves, high mitotic figures, and comedo necrosis were observed in addition to classic morphological features. More importantly, rare pseudopapillary structures were observed. The second case was of a 60-year-old woman in whom the tumor was mainly composed of multiple cysts filled with mucus. The remaining focal solid areas of the tumor comprised clear and acidophilic cells embedded in the hyalinizing stroma. Immunohistochemical analysis revealed that the tumor cells of both cases were positive for CK5/6, p40, and p63 expression, but negative for napsin A, TTF-1, and SOX10 expression. The HCCC diagnosis in both cases was validated by fluorescence in-situ hybridization (FISH) examination, which showed Ewing sarcoma breakpoint region 1-activating transcription factor 1 (EWSR1-ATF1) gene fusion. Primary pulmonary HCCC is a rare lung tumor originating from the bronchial mucosa, and its histological features may vary, such as rare pseudopapillary structures and abundant cysts. Thus, the diagnosis should be a combined analysis of histopathological characteristics with immunophenotype and molecular examination, including EWSR1-ATF1 gene fusion detection.

Deep learning based on carotid transverse B-mode scan videos for the diagnosis of carotid plaque: a prospective multicenter study.

OBJECTIVES: Accurate detection of carotid plaque using ultrasound (US) is essential for preventing stroke. However, the diagnostic performance of junior radiologists (with approximately 1 year of experience in carotid US evaluation) is relatively poor. We thus aim to develop a deep learning (DL) model based on US videos to improve junior radiologists' performance in plaque detection. METHODS: This multicenter prospective study was conducted at five hospitals. CaroNet-Dynamic automatically detected carotid plaque from carotid transverse US videos allowing clinical detection. Model performance was evaluated using expert annotations (with more than 10 years of experience in carotid US evaluation) as the ground truth. Model robustness was investigated on different plaque characteristics and US scanning systems. Furthermore, its clinical applicability was evaluated by comparing the junior radiologists' diagnoses with and without DL-model assistance. RESULTS: A total of 1647 videos from 825 patients were evaluated. The DL model yielded high performance with sensitivities of 87.03% and 94.17%, specificities of 82.07% and 74.04%, and areas under the receiver operating characteristic curve of 0.845 and 0.841 on the internal and multicenter external test sets, respectively. Moreover, no significant difference in performance was noted among different plaque characteristics and scanning systems. Using the DL model, the performance of the junior radiologists improved significantly, especially in terms of sensitivity (largest increase from 46.3 to 94.44%). CONCLUSIONS: The DL model based on US videos corresponding to real examinations showed robust performance for plaque detection and significantly improved the diagnostic performance of junior radiologists. KEY POINTS: • The deep learning model based on US videos conforming to real examinations showed robust performance for plaque detection. • Computer-aided diagnosis can significantly improve the diagnostic performance of junior radiologists in clinical practice.

A new species of Griseothosea (Lepidoptera, Limacodidae) from Wuyishan National Park, China.

A new species of Griseothosea nigrifasciata sp. nov. is described and illustrated from Wuyishan National Park, Fujian province, China. It differs from other members of Griseothosea by the male genitalia: uncus with strongly curved lateral margin, juxta with two finger-shaped protrusion apically, and aedeagus shorter and slenderer. The new species is supported molecularly by the analysis of a 658 bp fragment of the COI gene. A key to Griseothosea is provided.

Erratum: QUANHONG ZHANG, ALEXEY V. SOLOVYEV, JIAMIN LIANG, MIN WANG & HOUSHUAI WANG (2023) A new species of Griseothosea (Lepidoptera, Limacodidae) from Wuyishan National Park, China. Zootaxa, 5323 (3), 429434.

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PRLR and CACNA2D1 Impact the Prognosis of Breast Cancer by Regulating Tumor Immunity.

Phosphatase and tensin homolog (PTEN) is one of the highly susceptible genes to breast cancer (BC); however, the role of PTEN-related RNAs in BC remains poorly understood. Understanding the effect of PTEN-related RNAs and their mechanisms may be helpful to clinicians. We screened the differentially expressed RNAs (deRNAs) related to PTEN and established the competitive endogenous RNA (ceRNA) network by integrating several databases. After that, the RNA model, prolactin receptor (PRLR)/calcium voltage-gated channel auxiliary subunit alpha2delta 1 (CACNA2D1), was obtained by KM survival analysis and logistic regression analysis. Finally, mutation, methylation, functional enrichment, and immune correlation were analyzed to explore the roles of these RNAs. Our results showed that PRLR might be harmful to BC, while CACNA2D1 might be beneficial to BC. Furthermore, the abnormal expression of PRLR in BC might result from mutation and hypomethylation, while the aberrant expression of CACNA2D1 might be ascribed to methylation. Mechanistically, PRLR might affect the prognosis of BC by inhibiting the expression of immune checkpoints, while CACNA2D1 might improve the prognosis of BC by increasing the immune cells infiltrating into BC and up-regulating the expression of immune checkpoints. The abnormal expression of PRLR and CACNA2D1 in BC is closely related to the prognosis of BC, and they may serve as targets for the treatment of BC.

Two new species of the genus Microleon (Lepidoptera: Limacodidae) from China.

Two new species of the genus Microleon Butler, 1885, Microleon dianensis sp. nov. from Yunnan Province, China and Microleon simingensis sp. nov. from Zhejiang and Sichuan Provinces, China are described. Photographs of adults and male genitalia of these new species are provided, with a key to all currently known species of Microleon. The validity of new species is well supported by the molecular analysis of the fragment of mitochondrial cytochrome c oxidase subunit I (COI).

Evaluation of oocyte maturity using artificial intelligence quantification of follicle volume biomarker by three-dimensional ultrasound.

RESEARCH QUESTION: Can a novel deep learning-based follicle volume biomarker using three-dimensional ultrasound (3D-US) be established to aid in the assessment of oocyte maturity, timing of HCG administration and the individual prediction of ovarian hyper-response? DESIGN: A total of 515 IVF cases were enrolled, and 3D-US scanning was carried out on HCG administration day. A follicle volume biomarker established by means of a deep learning-based segmentation algorithm was used to calculate optimal leading follicle volume for predicting number of mature oocytes retrieved and optimizing HCG trigger timing. Performance of the novel biomarker cut-off value was compared with conventional two-dimensional ultrasound (2D-US) follicular diameter measurements in assessing oocyte retrieval outcome. Moreover, demographics, infertility work-up and ultrasound biomarkers were used to build models for predicting ovarian hyper-response. RESULTS: On the basis of the deep learning method, the optimal cut-off value of the follicle volume biomarker was determined to be 0.5 cm3 for predicting number of mature oocytes retrieved; its performance was significantly better than the conventional method (two-dimensional diameter measurement ≥10 mm). The cut-off value for leading follicle volume to optimize HCG trigger timing was determined to be 3.0 cm3 and was significantly associated with a higher number of mature oocytes retrieved (P = 0.01). Accuracy of the multi-layer perceptron model was better than two-dimensional diameter measurement (0.890 versus 0.785) and other multivariate classifiers in predicting ovarian hyper-response (P < 0.001). CONCLUSIONS: Deep learning segmentation methods and multivariate classifiers based on 3D-US were found to be potentially effective approaches for assessing mature oocyte retrieval outcome and individual prediction of ovarian hyper-response.