Krüppel-like factor 12 regulates aging ovarian granulosa cell apoptosis by repressing SPHK1 transcription and sphingosine-1-phosphate (S1P) production.

Oxidative stress triggered by aging, radiation, or inflammation impairs ovarian function by inducing granulosa cell (GC) apoptosis. However, the mechanism inducing GC apoptosis has not been characterized. Here, we found that ovarian GCs from aging patients showed increased oxidative stress, enhanced reactive oxygen species activity, and significantly decreased expression of the known antiapoptotic factor sphingosine-1-phosphate/sphingosine kinase 1 (SPHK1) in GCs. Interestingly, the expression of Krüppel-like factor 12 (KLF12) was significantly increased in the ovarian GCs of aging patients. Furthermore, we determined that KLF12 was significantly upregulated in hydrogen peroxide-treated GCs and a 3-nitropropionic acid-induced in vivo model of ovarian oxidative stress. This phenotype was further confirmed to result from inhibition of SPHK1 by KLF12. Interestingly, when endogenous KLF12 was knocked down, it rescued oxidative stress-induced apoptosis. Meanwhile, supplementation with SPHK1 partially reversed oxidative stress-induced apoptosis. However, this function was lost in SPHK1 with deletion of the binding region to the KLF12 promoter. SPHK1 reversed apoptosis caused by hydrogen peroxide-KLF12 overexpression, a result further confirmed in an in vitro ovarian culture model and an in vivo 3-nitropropionic acid-induced ovarian oxidative stress model. Overall, our study reveals that KLF12 is involved in regulating apoptosis induced by oxidative stress in aging ovarian GCs and that sphingosine-1-phosphate/SPHK1 can rescue GC apoptosis by interacting with KLF12 in negative feedback.

Zeaxanthin impairs angiogenesis and tumor growth of glioblastoma: An in vitro and in vivo study.

OBJECTIVES: To investigate the therapeutic effects of Zeaxanthin (Zea), one of the oxidized xanthophyll carotenoids belonging to the isoprenoids, on inhibiting the angiogenesis and tumor growth of glioblastoma (GBM) via an in vitro and in vivo study. METHODS: The effects of Zea on the proliferation, adhesion, migration and invasion of human GBM cell lines were detected by cell proliferation assay, cell adhesion assay and Transwell assay. The effect of Zea on angiogenesis was detected by rat aortic ring assay and human umbilical vein endothelial cells (HUVEC) in vitro tube formation assay. The effects of Zea on PARP, Caspase 3 and VEGFR2 phosphorylation as well as VEGFR2's downstream signaling pathway were detected by Western blot. The in vivo human GBM xenograft mouse model was employed to study the therapeutic efficacy of Zea. RESULTS: Zea impaired the proliferation, adhesion, migration and invasion of U87 and U251 cells as well as HUVECs. Rat aortic ring experiments displayed Zea significantly inhibited angiogenesis during VEGF-induced microvascular germination. In vitro and in vivo vascular experiments verified that Zea inhibited VEGF-induced HUVEC proliferation and capillary-like tube formation. Additionally, Zea induced GBM cells apoptosis via increasing the expression of cleaved PARP and Caspase 3. In HUVECs and U251 GBM cells, Zea down-regulated VEGF-induced activation of the VEGFR2 kinase pathway. Meanwhile the expression of p-AKT, p-ERK, p-STAT3 and FAK were all attenuated in U251 cells. Moreover, the effects of Zea on GBM cells proliferation could be blocked by VEGFR2 kinase inhibitor SU5408. These results suggest that Zea may hinder GBM angiogenesis and tumor growth through down-regulating a cascade of oncogenic signaling pathways, both through the inhibition of angiogenesis and the anti-tumor mechanism of a direct cytotoxic effect. Besides, Zea inhibits GBM angiogenesis and tumor growth exemplified through a xenograft mouse model in vivo. CONCLUSION: Zea impairs angiogenesis and tumor growth of GBM both in vitro and in vivo. It can be declared that Zea is a potential valuable anticancer candidate for the future treatment strategy of GBM.

APEX1 Knockdown Alleviates Inflammation and Fibrosis in Myocardial Infarction Through Promoting ZCCHC9 Expression and Blocking the p38 MAPK Signaling.

It was reported that serum apurinic/apyrimidinic endodeoxyribonuclease 1 (APEX1) level was higher in acute myocardial infarction (AMI) patients than in angina. This study aimed to investigate the role and mechanism of APEX1 in AMI progression. The mRNA and protein levels of APEX1 and zinc finger CCHC domain containing 9 (ZCCHC9) in blood specimens of AMI patients and normal controls were determined by RT-qPCR and Western blot assays, respectively. H9c2 cardiomyocytes were treated with angiotensin II (Ang II) to induce cardiomyocyte injury and then transfected with small interfering RNA against APEX1 (si-APEX1) or overexpression plasmids of ZCCHC9 (pcDNA-ZCCHC9). The cell viability, apoptosis, inflammatory cytokine levels, and fibrosis-associated protein expression in H9c2 cells were evaluated. ZCCHC9 promoter methylation were detected with methylation-specific PCR (MSP) assay. Then, rescue experiments were performed to explore whether APEX1 mediated cardiomyocyte functions by regulating ZCCHC9 expression. Furthermore, we explored whether the APEX1/ZCCHC9 axis regulated cardiomyocyte injury in AMI via the p38 MAPK signaling pathway. Additionally, an AMI rat model was established using the left anterior descending artery (LAD) ligation method and multipoint intramyocardial injection (5 points, 2 µL/point) of lentivirus (1 × 109 TU/mL) carrying scramble or si-APEX1 was conducted before modeling. The rats were euthanized four weeks after AMI modeling, and blood samples and myocardial tissues were harvested. The infarct area, cell apoptosis, inflammation, and fibrosis in myocardial tissues were detected. APEX1 was upregulated and ZCCHC9 was downregulated in blood samples of AMI patients compared with normal controls. APEX1 knockdown or ZCCHC9 overexpression attenuated Ang II-induced viability reduction, apoptosis, inflammation, and fibrosis in cardiomyocytes. APEX1 inhibited ZCCHC9 expression by promoting DNA methyltransferase 1 (DNMT1)-mediated ZCCHC9 promoter methylation. ZCCHC9 knockdown abolished the protective effects of APEX1 knockdown on Ang II-induced cardiomyocyte injury. APEX1 knockdown inhibited the p38 MAPK signal signaling, and anisomycin reversed the effect of APEX1 knockdown on cardiomyocyte functions. Additionally, APEX1 knockdown alleviated apoptosis, inflammation, and fibrosis in myocardial tissues of AMI rats. APEX1 knockdown attenuated Ang II-induced apoptosis, inflammation, and fibrosis in cardiomyocytes although promoting ZCCHC9 expression and inhibiting the p38 MAPK signaling pathway, thus relieving myocardial infarction, inflammation, and fibrosis in AMI rats.

Observation on Short-Term Therapeutic Effect of Electroacupuncture on Compensatory Hypertrophy Of Gastrocnemius Muscle: A Retrospective Cohort Study.

Objective: In Oriental women, having thick and short legs is a common posture problem, and having an imperfect lower leg shape is one of the factors that can cause feelings of inferiority. To address this issue, a retrospective cohort study was conducted to investigate the effectiveness of electroacupuncture in improving compensatory hypertrophy of the gastrocnemius muscle while also reducing side effects. The goal of this study is to improve the clinical treatment plan for this type of problem. Methods: This retrospective cohort study was conducted between December 2020 and March 2022 at the Changhai Hospital of Shanghai, Shanghai, China. This study included 80 patients who were divided into two equal groups - the infrared (IR) group and the electroacupuncture (EA) group, based on the type of treatment they received during the research. The EA group received electroacupuncture and infrared treatment, while the IR group used an infrared therapeutic instrument to irradiate their lower legs on both sides. The main outcome measures were 3 calf circumference levels and the cross-sectional area of the gastrocnemius muscle in two-dimensional ultrasound. The secondary outcome measure was the incidence of adverse events. Results: According to the data, before the treatment there were no statistically significant differences between the two groups in calf circumference and ultrasound gastrocnemius cross-sectional area. After the treatment, the value of each calf circumference level and ultrasound gastrocnemius cross-sectional area were significantly lower than the value collected before the treatment in the EA Group. However, there is no significant change in the data of the infrared therapeutic group before and after treatment. By comparing the data between the 2 groups we collected after the treatment, the value of each calf circumference level and ultrasound gastrocnemius cross-sectional area of the EA group is significantly lower than that of the IR group. Only 8 patients suffered from lower limb pain and other discomfort after treatment, and these symptoms did not cause dissatisfaction. Conclusion: Electroacupuncture is an effective treatment for compensatory hypertrophy of the gastrocnemius muscle.

Candidate Gene Identification and Transcriptome Analysis of Tomato male sterile-30 and Functional Marker Development for ms-30 and Its Alleles, ms-33, 7B-1, and stamenless-2.

Male sterility is a valuable trait for hybrid seed production in tomato (Solanum lycopersicum). The mutants male sterile-30 (ms-30) and ms-33 of tomato exhibit twisted stamens, exposed stigmas, and complete male sterility, thus holding potential for application in hybrid seed production. In this study, the ms-30 and ms-33 loci were fine-mapped to 53.3 kb and 111.2 kb intervals, respectively. Tomato PISTILLATA (TPI, syn. SlGLO2), a B-class MADS-box transcription factor gene, was identified as the most likely candidate gene for both loci. TPI is also the candidate gene of tomato male sterile mutant 7B-1 and sl-2. Allelism tests revealed that ms-30, ms-33, 7B-1, and sl-2 were allelic. Sequencing analysis showed sequence alterations in the TPI gene in all these mutants, with ms-30 exhibiting a transversion (G to T) that resulted in a missense mutation (S to I); ms-33 showing a transition (A to T) that led to alternative splicing, resulting in a loss of 46 amino acids in protein; and 7B-1 and sl-2 mutants showing the insertion of an approximately 4.8 kb retrotransposon. On the basis of these sequence alterations, a Kompetitive Allele Specific PCR marker, a sequencing marker, and an Insertion/Deletion marker were developed. Phenotypic analysis of the TPI gene-edited mutants and allelism tests indicated that the gene TPI is responsible for ms-30 and its alleles. Transcriptome analysis of ms-30 and quantitative RT-PCR revealed some differentially expressed genes associated with stamen and carpel development. These findings will aid in the marker-assisted selection for ms-30 and its alleles in tomato breeding and support the functional analysis of the TPI gene.

Optical absorption enhancement in inhomogeneous InGaN nanowire arrays photocathode.

In the development of surface structures, nanowire arrays (NWAS) have been widely studied because of their trapping effect. In this paper, the finite difference time domain (FDTD) method is used to simulate homogeneous and inhomogeneous NWAS. We studied the influence of the structural parameters of InGaN NWAS and inhomogeneous arrays on optical response properties. The optical response includes light absorptivity and cutoff wavelength sensitivity. The simulation results show that the inhomogeneous NWAS can increase the effective transmission distance of light on the surface, thus greatly improving the optical absorption capacity of InGaN NWAS. We can obtain high sensitivity of cut-off wavelength by adjusting the structural parameters of the side nanowires. We find that by reducing the diameters and heights of the side nanowires, a higher light absorption rate can be obtained, which is a 5% improvement compared to uniform NWAS. Therefore, the research in this paper can provide some theoretical reference for the experiment and preparation of InGaN photocathodes.

Antitumor activity of mianserin (a tetracyclic antidepressant) primarily driven by the inhibition of SLC1A5-mediated glutamine transport.

Targeting tumor metabolic vulnerabilities such as "glutamine addiction" has become an attractive approach for the discovery of novel antitumor agents. Among various mechanisms explored, SLC1A5, a membrane transporter that plays an important role in glutamine cellular uptake, represents a viable target to interfere with tumor's ability to acquire critical nutrients during proliferation. In the present study, a stably transfected HEK293 cell line with human SLC1A5 (HEK293-SLC1A5) was established for the screening and identification of small molecule SLC1A5 inhibitors. This in vitro system, in conjunction with direct measurement of SLC1A5-mediated L-glutamine-2,3,3,4,4-D5 (substrate) uptake, was practical and efficient in ensuring the specificity of SLC1A5 inhibition. Among a group of diverse compounds tested, mianserin (a tetracyclic antidepressant) demonstrated a marked inhibition of SLC1A5-mediated glutamine uptake. Subsequent investigations using SW480 cells demonstrated that mianserin was capable of inhibiting SW480 tumor growth both in vitro and in vivo, and the in vivo antitumor efficacy was correlated to the reduction of glutamine concentrations in tumor tissues. Computational analysis revealed that hydrophobic interactions between SLC1A5 and its inhibitors could be a critical factor in drug design. Taken together, the current findings confirmed the feasibility of targeting SLC1A5-mediated glutamine uptake as a novel approach for antitumor intervention. It is anticipated that structural insights obtained based on homology modeling would lead to the discovery of more potent and specific SLC1A5 inhibitors for clinical development.

Reward Weakened Inhibition of Return (IOR) in the Near Depth Plane.

In attentional orienting, researchers have proposed that reward history is a component of attentional control, as the reward value might enhance the spatial attention process to achieve more efficient goal-directed behavior and to improve target-detection performance. Although the effect of reward-induced motivation on attentional orienting has been studied in two-dimensional (2-D) space, the specific mechanisms underlying the influence of reward on inhibition of return (IOR) of attentional orienting in three-dimensional space (3-D) remain unclear. In the present study, by incorporating the Posner spatial-cueing paradigm into a virtual 3-D environment, we aimed to investigate the influence of reward on IOR in 3-D space. The results showed the following: (1) IOR size in the rewarded conditions was smaller than IOR size in the unrewarded condition in the near depth plane, resulting in an IOR difference with or without reward. (2) Reward weakened IOR in the near depth plane because the response to the uncued location was delayed, not because the response to the cued location was accelerated. The present study indicated that the different depth planes of the target location in 3-D space could influence the interaction between reward and IOR, and reward weakened IOR in the near depth plane.

Deep-Blue Electroluminescence of Perovskites with Reduced Dimensionality Achieved by Manipulating Adsorption-Energy Differences.

The lagging development of deep-blue perovskite light-emitting diodes (PeLEDs) heavily impedes their practical applications in full-color display due to the absence of spectrally stable emitters and the mismatch of carrier injection capacity. Herein, we report highly efficient deep-blue PeLEDs through a new chemical strategy that addresses the dilemma for simultaneously constant electroluminescence (EL) spectra and high-purify phase in reduced-dimensional perovskites. The success lies in the control of adsorption-energy differences between phenylbutylamine (PBA) and ethylamine (EA) interacting with perovskites, which facilitates narrow n-value distribution. This approach leads to an increased exciton binding energy and enhanced surface potential, hence improving radiative recombination. As a result, an external quantum efficiency of 4.62 % is achieved in PeLEDs with a stable EL peak at 457 nm, demonstrating the best reported result for deep-blue PeLEDs so far.

Comparative transcriptomic analysis reveals the mechanistic basis of Pib-mediated broad spectrum resistance against Magnaporthe oryzae.

Rice blast, caused by the fungus Magnaporthe oryzae, is a highly damaging disease. Introducing genes, which confer a broad spectrum resistance to the disease, such as Pib, makes an important contribution to protecting rice production. However, little is known regarding the mechanistic basis of the products of such genes. In this study, transcriptome of the cultivar Lijiangxintuanheigu (LTH) and its monogenic IRBLb-B which harbors Pib treated with M. oryzae were compared. Among the many genes responding transcriptionally to infection were some encoding products involved in the metabolism of ROS (reactive oxygen species), in jasmonate (JA) metabolism, and WRKY transcription factors, receptor kinases, and resistance response signal modulation. The down-regulation of genes encoding peroxiredoxin and glutathione S transferases implied that the redox homeostasis is essential for the expression of Pib-mediated resistance. The up-regulation of seven disease resistance-related genes, including three encoding a NBS-LRR protein, indicated that disease resistance-related genes are likely tend to support the expression of Pib resistance. These data revealed that potential candidate genes and transcriptional reprogramming were involved in Pib-mediated resistance mechanisms.

Enhancement of photoemission capability and electron collection efficiency of field-assisted GaN nanowire array photocathode.

GaN has interesting prospects in applications for spectrum-tunable solid-state devices with photoelectric conversion function. Similarly, single nanowires or nanowire arrays (NWAs) proceed to exhibit good photon absorbance and photoemission characteristics as vacuum devices based on the external photoelectric effect. However, the collection of photoelectrons emitted from a nanowire surface has become the greatest impediment to the progress of GaN NWAs photocathodes. In this study, a field-assisted GaN NWA photocathode is proposed. The photoemission efficiency and electron collection efficiency of the field-assisted GaN NWA photocathode are derived. The results suggest that the external field can effectively enhance the photoemission capacity and electron collection efficiency of the photocathode. Based on the theoretical model, the structural parameters of NWAs and the field intensity are optimized. When the field intensity is 1 V µm-1, the collected photocurrent of the GaN NWA photocathode reaches a maximum. For NWAs with an aspect ratio of 1:1, the optimal incident angle of light is 70°. This study provides a theoretical guide for the incorporation of an external field in a GaN NWA photocathode with the purpose of enhancing photoemission and electron collection capacity.

Amorphous Metal-Free Room-Temperature Phosphorescent Small Molecules with Multicolor Photoluminescence via a Host-Guest and Dual-Emission Strategy.

Metal-free room-temperature phosphorescence (RTP) materials offer unprecedented potentials for photoelectric and biochemical materials due to their unique advantages of long lifetime and low toxicity. However, the achievements of phosphorescence at ambient condition so far have been mainly focused on ordered crystal lattice or on embedding into rigid matrices, where the preparation process might bring out poor repeatability and limited application. In this research, a series of amorphous organic small molecular compounds were developed with efficient RTP emission through conveniently modifying phosphor moieties to ß-cyclodextrin (ß-CD). The hydrogen bonding between the cyclodextrin derivatives immobilizes the phosphors to suppress the nonradiative relaxation and shields phosphors from quenchers, which enables such molecules to emit efficient RTP emission with decent quantum yields. Furthermore, one such cyclodextrin derivative was utilized to construct a host-guest system incorporating a fluorescent guest molecule, exhibiting excellent RTP-fluorescence dual-emission properties and multicolor emission with a wide range from yellow to purple including white-light emission. This innovative and universal strategy opens up new research paths to construct amorphous metal-free small molecular RTP materials and to design organic white-light-emitting materials using a single supramolecular platform.

Defect-controlled transverse localization of light in triangular optical lattices.

We present a detailed analysis of the transverse localization of light in a triangular optical lattice with a negative defect of different depths and positions. The analysis allows us to explore some essentials of the localization behavior. We characterize the localization strength by global propagation of the signal beam and its properties on the output plane. It is demonstrated that the enhancement of the localization is dependent on the defect depth. Additionally, the calculation of the ray trace of the signal beam shows that the position of the defect has a crucial influence on the localization.

Effects of 10 KM run on foot morphology and bilateral symmetry in male recreational runners.

Foot morphology and arch integrity do not remain constant during a running bout. Previous studies have reported inconsistent changes in foot sizes and arch parameters and this discrepancy may be related to the variation in their test duration, e.g., 15-min treadmill run vs. 30 KM trial. Hence, this study sought to evaluate the change in foot morphology, arch integrity and bilateral symmetry after a 10 KM run among 19 male recreational runners. Before and after the run, a portable foot scanner was used to capture the 3-dimensional foot images and measure foot dimensions in both weight-bearing and non-weight-bearing conditions. Foot arch integrity was quantified by arch height ratio, arch height index, and arch stiffness index (ASI). Bilateral symmetry was evaluated by calculating the symmetry index. Increased foot length (p = 0.007; η p 2 = 0.18) and decreased ball girth (p = 0.038; η p 2 = 0.11) were demonstrated following the run with absolute differences of less than 2 mm. Navicular height, dorsum height, arch height ratio and arch height index significantly decreased after the run (p < 0.001; η p 2 ≥0.30) whereas ASI increased (p < 0.001, η p 2 = 0.33) and navicular height drop reduced (p < 0.001, η p 2 = 0.37). Significances of symmetry index were only demonstrated for navicular height (p = 0.019, effect size = 0.37) and arch height ratio (p = 0.019, effect size = 0.42). A few changes in foot morphology were detected but a reduction in foot arch height was demonstrated, which may give shoe manufacturers insights into shoe design. Male recreational runners were recommended to choose shoes with arch support for maintaining foot arch function during a 10 KM run.

Disruption of LPA-LPAR1 pathway results in lung tumor growth inhibition by downregulating B7-H3 expression in fibroblasts.

BACKGROUND: Lysophosphatidic acids (LPAs) belong to a class of bioactive lysophospholipids with multiple functions including immunomodulatory roles in tumor microenvironment (TME). LPA exerts its biological effects via its receptors that are highly expressed in fibroblasts among other cell types. As cancer-associated fibroblasts (CAFs) are a key component of the TME, it is important to understand LPA signaling and regulation of receptors in fibroblasts or CAFs and associated regulatory roles on immunomodulation-related molecules. METHODS: Cluster analysis, immunoblotting, real-time quantitative-PCR, CRISPR-Cas9 gene editing system, immunohistochemical staining, coculture model, and in vivo xenograft model were used to investigate the effects of LPA-LPAR1 on B7-H3 in tumor promotion of CAFs. RESULTS: In this study, we found that LPAR1 and CD276 (B7-H3) were generally highly expressed in fibroblasts with good expression correlation. LPA induced B7-H3 up-expression through LPAR1, and stimulated fibroblasts proliferation that could be inhibited by silencing LPAR1 or B7-H3 as well as small molecule LPAR1 antagonist (Ki16425). Using engineered fibroblasts and non-small cell lung carcinoma (NSCLC) cell lines, subsequent investigations demonstrated that CAFs promoted the proliferation of NSCLC in vitro and in vivo, and such effect could be inhibited by knocking out LPAR1 or B7-H3. CONCLUSION: The present study provided new insights for roles of LPA in CAFs, which could lead to the development of innovative therapies targeting CAFs in the TME. It is also reasonable to postulate a combinatory approach to treat malignant fibrous tumors (such as NSCLC) with LPAR1 antagonists and B7-H3 targeting therapies.

The pro-tumor activity of INTS7 on lung adenocarcinoma via inhibiting immune infiltration and activating p38MAPK pathway.

Lung adenocarcinoma (LUAD) is the most common lung cancer, accounting for 19.4% of all cancer deaths. Our previous study discovered that INTS7 expression was upregulated in LUAD, while the precise mechanism by which INTS7 exerts pro-cancer effects remains unknown. In our study, shRNA was used to knockdown the expression of INTS7 in A549 cells. Cancer behaviors in vitro were determined by CCK8 and transwell assays. Xenograft mice models were constructed to detect the tumorigenesis in vivo. Immunofluorescence and toluidine blue staining were used to test the immune infiltration. Bioinformatics analysis was adopted to predict the potential signaling pathways and construct INTS7-derived genomic prognostic model. Western blot was utilized to confirm the molecular pathways. In total, downregulation of INTS7 suppressed proliferation, invasion and migration of A549 cells, as well as tumor growth. Bioinformatics and western blot analysis indicated that p38MAPK pathway participated in the regulatory mechanism of INTS7. Moreover, INTS7 expression was negatively correlated with infiltration of memory B cells and mast cells, while positively correlated with infiltration of macrophages M2. A nomogram, including INTS7-derived risk score, was used to estimate individual's survival probability. Generally, our findings provided comprehensive understanding of the molecular mechanisms about INTS7, and targeting INTS7 may represent a potential therapy for LUAD.

Primary Budd-Chiari syndrome versus sinusoidal obstruction syndrome: a review.

Budd-Chiari syndrome (BCS) and sinusoidal obstruction syndrome (SOS) are two major vascular disorders of the liver, of which both can cause portal hypertension related complications, but their locations of obstruction are different. BCS refers to the obstruction from the hepatic vein to the junction between the inferior vena cava and right atrium, which is the major etiology of post-sinusoidal portal hypertension; by comparison, SOS is characterized as the obstruction at the level of hepatic sinusoids and terminal venulae, which is a cause of sinusoidal portal hypertension. Both of them can cause hepatic congestion with life-threatening complications, especially acute liver failure and chronic portal hypertension, and share some similar features in terms of imaging and clinical presentations, but they have heterogeneous risk factors, management strategy, and prognosis. Herein, this paper reviews the current evidence and then summarizes the difference between primary BCS and SOS in terms of risk factors, clinical features, diagnosis, and treatment.

Peimine induces apoptosis of glioblastoma cells through regulation of the PI3K/AKT signaling pathway.

Glioblastoma (GBM) is one of the most malignant forms of intracranial tumors, with high mortality rates and invariably poor prognosis, due to the limited clinical treatment strategies available. As a natural compound, peimine's favorable pharmacological activities have been widely revealed. However, potential inhibitory effects of peimine on GBM have not been explored. In the present study, both in vitro and in vivo experiments were performed to elucidate the effects of peimine on GBM and to further delineate the underlying molecular mechanism of action. Different doses (0, 25 and 50 µM) of peimine were added to U87 cells, before MTT, colony formation, wound healing, Transwell migration and invasion, reactive oxygen species and mitochondrial transmembrane potential assays were used to measure proliferation, migration, invasion and apoptosis. Furthermore, western blotting was used to examine the possible effects of peimine on the expression of proteins associated with apoptosis and the PI3K/AKT signaling pathway. Subsequently, a GBM mouse xenograft model was used to assess the effects of peimine in vivo. The findings showed that peimine inhibited GBM proliferation, migration and invasion in a dose-dependent manner, whilst also inducing apoptosis. Peimine also reduced tumor growth in vivo. Mechanistically, peimine downregulated the expression of Bcl-2 and Caspase 3, whilst upregulating the protein expression levels of p53, Bax and Cleaved-Caspase 3 in a dose-dependent manner. In addition, PI3K and AKT phosphorylation levels were found to be decreased by peimine in a dose-dependent manner. In conclusion, these findings suggest that peimine may limit GBM growth by regulating the PI3K/AKT signaling pathway both in vitro and in vivo. These findings may have promising clinical implications.

ZP1-Y262C mutation causes abnormal zona pellucida formation and female infertility in humans.

Defective oocyte maturation is a common cause of female infertility. The loss of the zona pellucida (ZP) represents a specific condition of impaired oocyte maturation. The extracellular matrix known as the ZP envelops mammalian oocytes and preimplantation embryos, exerting significant influence on oogenesis, fertilization, and embryo implantation. However, the genetic factors leading to the loss of the ZP in oocytes are not well understood. This study focused on patients who underwent oocyte retrieval surgery after ovarian stimulation and were found to have abnormal oocyte maturation without the presence of the ZP. Ultrasonography was performed during the surgical procedure to evaluate follicle development. Peripheral blood samples from the patient were subjected to exome sequencing. Here, a novel, previously unreported heterozygous mutation in the ZP1 gene was identified. Within the ZP1 gene, we discovered a novel heterozygous mutation (ZP1 NM_207341.4:c.785A>G (p.Y262C)), specifically located in the trefoil domain. Bioinformatics comparisons further revealed conservation of the ZP1-Y262C mutation across different species. Model predictions of amino acid mutations on protein structure and cell immunofluorescence/western blot experiments collectively confirmed the detrimental effects of the ZP1-Y262C mutation on the function and expression of the ZP1 protein. The ZP1-Y262C mutation represents the novel mutation in the trefoil domain of the ZP1 protein, which is associated with defective oocyte maturation in humans. Our report enhances comprehension regarding the involvement of ZP-associated genes in female infertility and offers enriched understanding for the genetic diagnosis of this condition.

OsLESV and OsESV1 promote transitory and storage starch biosynthesis to determine rice grain quality and yield.

Transitory starch is an important carbon source in leaves, and its biosynthesis and metabolism are closely related to grain quality and yield. The molecular mechanisms controlling leaf transitory starch biosynthesis and degradation and their effects on rice (Oryza sativa) quality and yield remain unclear. Here, we show that OsLESV and OsESV1, the rice orthologs of AtLESV and AtESV1, are associated with transitory starch biosynthesis in rice. The total starch and amylose contents in leaves and endosperms are significantly reduced, and the final grain quality and yield are compromised in oslesv and osesv1 single and oslesv esv1 double mutants. Furthermore, we found that OsLESV and OsESV1 bind to starch, and this binding depends on a highly conserved C-terminal tryptophan-rich region that acts as a starch-binding domain. Importantly, OsLESV and OsESV1 also interact with the key enzymes of starch biosynthesis, granule-bound starch synthase I (GBSSI), GBSSII, and pyruvate orthophosphote dikiase (PPDKB), to maintain their protein stability and activity. OsLESV and OsESV1 also facilitate the targeting of GBSSI and GBSSII from plastid stroma to starch granules. Overexpression of GBSSI, GBSSII, and PPDKB can partly rescue the phenotypic defects of the oslesv and osesv1 mutants. Thus, we demonstrate that OsLESV and OsESV1 play a key role in regulating the biosynthesis of both leaf transitory starch and endosperm storage starch in rice. These findings deepen our understanding of the molecular mechanisms underlying transitory starch biosynthesis in rice leaves and reveal how the transitory starch metabolism affects rice grain quality and yield, providing useful information for the genetic improvement of rice grain quality and yield.