[Notch1/Akt/Foxo1 Pathway Regulated by Kisspeptin Is Involved in Endometrial Decidualization in Patients With Recurrent Spontaneous Abortion]. / kisspeptin调控Notch1/Akt/Foxo1é€šè·¯å‚ä¸Žå¤å‘æ€§æµäº§æ‚£è€ å­å®«å† è†œèœ•è†œåŒ–.

Objective: Kisspeptin, a protein encoded by the KISS1 gene, functions as an essential factor in suppressing tumor growth. The intricate orchestration of cellular processes such as proliferation and differentiation is governed by the Notch1/Akt/Foxo1 signaling pathway, which assumes a central role in maintaining cellular homeostasis. In the specific context of this investigation, the focal point lies in a meticulous exploration of the intricate mechanisms underlying the regulatory effect of kisspeptin on the process of endometrial decidualization. This investigation delves into the interplay between kisspeptin and the Notch1/Akt/Foxo1 signaling pathway, aiming to elucidate its significance in the pathophysiology of recurrent spontaneous abortion (RSA). Methods: We enrolled a cohort comprising 45 individuals diagnosed with RSA, who were admitted to the outpatient clinic of the Reproductive Center at the Second Affiliated Hospital of Soochow University between June 2020 and December 2020. On the other hand, an additional group of 50 women undergoing elective abortion at the outpatient clinic of the Family Planning Department during the same timeframe was also included. To comprehensively assess the molecular landscape, Western blot and RT-qPCR were performed to analyze the expression levels of kisspeptin (and its gene KISS1), IGFBP1 (an established marker of decidualization), Notch1, Akt, and Foxo1 within the decidua. Human endometrial stromal cells (hESC) were given targeted interventions, including treatment with siRNA to disrupt KISS1 or exposure to kisspeptin10 (the bioactive fragment of kisspeptin), and were subsequently designated as the siKP group or the KP10 group, respectively. A control group comprised hESC was transfected with blank siRNA, and cell proliferation was meticulously evaluated with CCK8 assay. Following in vitro induction for decidualization across the three experimental groups, immunofluorescence assay was performed to identify differences in Notch1 expression and decidualization morphology between the siKP and the KP10 groups. Furthermore, RT-qPCR and Western blot were performed to gauge the expression levels of IGFBP1, Notch1, Akt, and Foxo1 across the three cell groups. Subsequently, decidualization was induced in hESC by adding inhibitors targeting Notch1, Akt, and Foxo1. The expression profiles of the aforementioned proteins and genes in the four groups were then examined, with hESC induced for decidualization without adding inhibitors serving as the normal control group. To establish murine models of normal pregnancy (NP) and RSA, CBA/J×BALB/c and CBA/J×DBA/2 mice were used. The mice were respectively labeled as the NP model and RSA model. The experimental groups received intraperitoneal injections of kisspeptin10 and kisspeptin234 (acting as a blocker) and were designated as RSA-KP10 and NP-KP234 groups. On the other hand, the control groups received intraperitoneal injections of normal saline (NS) and were referred to as RSA-NS and NP-NS groups. Each group comprised 6 mice, and uterine tissues from embryos at 9.5 days of gestation were meticulously collected for observation of embryo absorption and examination of the expression of the aforementioned proteins and genes. Results: The analysis revealed that the expression levels of kisspeptin, IGFBP1, Notch1, Akt, and Foxo1 were significantly lower in patients diagnosed with RSA compared to those in women with NP (P<0.01 for kisspeptin and P<0.05 for IGFBP1, Notch1, Akt, and Foxo1). After the introduction of kisspeptin10 to hESC, there was an observed enhancement in decidualization capability. Subsequently, the expression levels of Notch1, Akt, and Foxo1 showed an increase, but they decreased after interference with KISS1. Through immunofluorescence analysis, it was observed that proliferative hESC displayed a slender morphology, but they transitioned to a rounder and larger morphology post-decidualization. Concurrently, the expression of Notch1 increased, suggesting enhanced decidualization upon the administration of kisspeptin10, but the expression decreased after interference with KISS1. Further experimentation involved treating hESC with inhibitors specific to Notch1, Akt, and Foxo1 separately, revealing a regulatory sequence of Notch1/Akt/Foxo1 (P<0.05). In comparison to the NS group, NP mice administered with kisspeptin234 exhibited increased fetal absorption rates (P<0.001) and decreased expression of IGFBP1, Notch1, Akt, and Foxo1 (P<0.05). Conversely, RSA mice administered with kisspeptin10 demonstrated decreased fetal absorption rates (P<0.001) and increased expression levels of the aforementioned molecules (P<0.05). Conclusion: It is suggested that kisspeptin might exert its regulatory influence on the process of decidualization through the modulation of the Notch1/Akt/Foxo1 signaling cascade. A down-regulation of the expression levels of kisspeptin could result in suboptimal decidualization, which in turn might contribute to the development or progression of RSA.

The evolution of atomistic structure and mechanical property of coke in the gasification process with CO2 and H2O at different temperatures: A ReaxFF molecular dynamics study.

CONTEXT: An atomistic coke carbon model was constructed to simulate the structural evolution in the gasification and stretching process. The coke model was placed in a box with different CO2/H2O content to investigate the evolution of the atomistic structure of coke during the gasification. It was found that different atmospheric concentrations had different effects on the structure and reaction sites of the coke model. The CO2 molecules tended to dissolve on the surface of coke and disrupt its surface structure, while H2O molecules were more likely to enter the coke model to disrupt the internal structure. For tensile simulation, it was found that CO2 and H2O had different effects on the tensile resistance of the coke model. Controlling the composition content of the reaction gas can effectively influence the tensile strength of the coke model. By revealing the behavior of coke model at the micro scale, it provides a theoretical basis for the industrial coke application process. METHODS: Large-scale Atomic/Molecular Massively Parallel Simulator (LAMMPS) was used to conduct the molecular dynamics using the reactive force field (ReaxFF). The atomistic model of coke carbon was constructed using the well-known annealing and quenching method, and its composition is determined according to the element analysis of industrial coke. The structural evolution in the gasification with CO2/H2O and the stretching process were analyzed in detail. Molecular dynamics simulations with reactive force field (ReaxFF-MD) were used to simulate the coke dissolution reaction under CO2/H2O atmosphere and the coke stretching process. The atmosphere ratio was modified to investigate the changes in coke structure under different atmosphere conditions. The Packmol software was used to place gas and coke models into the same box. During the reaction process, the Ovito software was used to perform corresponding visualization analysis on the changes in the atomic structure of coke.

Effect of Different sp2 Bond Contents on the Reactivity and Mechanical Properties of Coke Carbon: A ReaxFF Molecular Dynamics Study.

In this study, ReaxFF-MD was used to construct a large-molecule model of coke containing 3000 atoms, and the sp2 bond content of the model was controlled by changing the heating and cooling rates. The increase of the sp2 bond content led to a significant difference in the reactivity of coke. The presence of the sp2 bond caused the carbon atoms inside the coke to change into a circular structure, making it more difficult for the gaseous atoms to adsorb on the surface of the coke. It significantly reduced the gasification reaction rate of coke in the CO2 and H2O atmospheres. In the tensile simulation experiment, it was found that the stretching process of coke was mainly divided into three stages: an elastic stretching stage, a plastic stretching stage, and a model fracture stage. During the stretching process, the carbon ring structure would undergo a C-C bond fracture while generating carbon chains to resist stress. The results indicated that the presence of sp2 bonds can effectively reduce the phenomenon of excessive local stress on coke to improve its tensile resistance. The method developed in this paper may provide further ideas and platforms for the research on coke performance.

Phosphohistidine signaling promotes FAK-RB1 interaction and growth factor-independent proliferation of esophageal squamous cell carcinoma.

Current clinical therapies targeting receptor tyrosine kinases including focal adhesion kinase (FAK) have had limited or no effect on esophageal squamous cell carcinoma (ESCC). Unlike esophageal adenocarcinomas, ESCC acquire glucose in excess of their anabolic need. We recently reported that glucose-induced growth factor-independent proliferation requires the phosphorylation of FAKHis58. Here, we confirm His58 phosphorylation in FAK immunoprecipitates of glucose-stimulated, serum-starved ESCC cells using antibodies specific for 3-phosphohistidine and mass spectrometry. We also confirm a role for the histidine kinase, NME1, in glucose-induced FAKpoHis58 and ESCC cell proliferation, correlating with increased levels of NME1 in ESCC tumors versus normal esophageal tissues. Unbiased screening identified glucose-induced retinoblastoma transcriptional corepressor 1 (RB1) binding to FAK, mediated through a "LxCxE" RB1-binding motif in FAK's FERM domain. Importantly, in the absence of growth factors, glucose increased FAK scaffolding of RB1 in the cytoplasm, correlating with increased ESCC G1→S phase transition. Our data strongly suggest that this glucose-mediated mitogenic pathway is novel and represents a unique targetable opportunity in ESCC.

Density Functional Studies on the Atomistic Structure and Properties of Iron Oxides: A Parametric Study.

With the aim to find the best simulation routine to accurately predict the ground-state structures and properties of iron oxides (hematite, magnetite, and wustite) using density functional theory (DFT) with Hubbard-U correction, a significant amount of DFT calculations were conducted to investigate the influence of various simulation parameters (energy cutoff, K-point, U value, magnetization setting, smearing value, etc.) and pseudopotentials on the structures and properties of iron oxides. With optimized simulation parameters, the obtained equation of state, lattice constant, bulk moduli, and band gap is much closer to the experimental values compared with previous studies. Due to the strong coupling between the 2p orbital of O and the 3d orbital of Fe, it was found that Hubbard-U correction obviously improved the results for all three kinds of iron oxides including magnetite which has not yet been tested with U correction before, but the U value should be different for different oxides (3 ev, 4 ev, 4 ev for hematite, magnetite, and wustite, respectively). Two kinds of spin magnetism settings for FeO are considered, which should be chosen according to different calculation purposes. The detailed relationship between the parameter settings and the atomic structures and properties were analyzed, and the general principles for future DFT calculation of iron oxides were provided.

Effect of Food on the Pharmacokinetics of Senaparib (IMP4297) in Healthy Chinese Subjects.

BACKGROUND AND OBJECTIVES: Data on the effect of food on the pharmacokinetics of senaparib (previously IMP4297), an oral poly (adenosine diphosphate-ribose) polymerase inhibitor, are limited. This study was conducted to evaluate the effect of food on the pharmacokinetics of senaparib in healthy Chinese subjects. METHODS: This is a phase I, open-label, randomized, single-dose, two-way crossover study. Healthy Chinese male subjects were randomized 1:1 to receive a single dose of senaparib 100 mg in two prandial states: fasted or after a high-fat meal; subjects were given a second dose after switching prandial states and a washout period of at least 7 days. Pharmacokinetics were assessed at pre-dose and up to 72 h post-dose. Safety was assessed throughout the study. RESULTS: Sixteen subjects were randomized and included in the pharmacokinetic analysis; 15 completed the study. The presence of food slowed the rate of senaparib absorption (time to maximum concentration) by ~ 3 h and reduced the maximum concentration of senaparib by ~ 24%. Total exposure to senaparib was higher in the fed than fasted state; senaparib area under the plasma concentration-time curve from time zero to the last measurable concentration and area under the plasma concentration-time curve from time zero to infinity were increased by ~ 24 and ~28%, respectively. Safety profiles were similar in both prandial states. All treatment-emergent adverse events were grade 1 in severity; no serious adverse events or deaths were reported. CONCLUSIONS: Food slightly decreased the rate and increased the extent of senaparib absorption following oral administration. However, the effect of food on various exposure parameters was not considered clinically meaningful. Safety data were consistent with the known profile of senaparib and senaparib was well tolerated in the fed and fasted states in healthy subjects. These results indicated that senaparib could be administered orally with or without food. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov NCT04057729.

Prediction of Cervical Lymph Nodes Metastasis in Papillary Thyroid Carcinoma (PTC) Using Nodal Staging Score (NSS).

Background: Cervical lymph node metastasis is commonly seen in papillary thyroid carcinoma. Surgery is the preferred treatment for PTC with cervical lymph node metastasis. There is no alternate ultrasound, neck CT, and thyroglobulin (Tg) methods to assess the occult lymph node metastasis. For moderate-and high-risk PTC, the number of lymph nodes to be dissected should be increased to remove the occult lymph node metastasis. Objective: This study was designed to develop a nodal staging score model to predict the likelihood of lymph node metastasis in papillary thyroid carcinoma (PTC), and further guide the treatments. Material and Methods. Data were collected from the SEER database. Patients with PTC from 2000 to 2005 were selected. The beta-binomial model was adopted to establish a nodal staging score (NSS)-based model. The NSS-based model was built according to gender, age, extrathyroidal invasion, tumor multifocality, tumor size, and T stage of the patients. A total of 12,431 PTC patients were included in our study. Various types of lymph nodes were examined based on various categories (incidence, risk assessment) to evaluate the results. Results: 5,959 (47.9%) patients in the study were positive and 6,472 (52.1%) were confirmed negative for lymph node metastasis. The corrected incidence of lymph node metastasis was higher than that of direct calculation, regardless of the factors that affected lymph node metastasis. There were significant differences in the OS of PTC patients among the four groups and T stage (p is less than 0.05), indicating that cervical lymph node metastasis would have an impact on the prognosis of patients. Conclusion: In conclusion, an NSS-based model base on a variety of clinicopathological factors can be used to predict lymph node metastasis. It is important to evaluate the risk of occult lymph node metastasis in the treatment of PTC.. Since, this statistical model can describe the risk of occult lymph node metastasis in patients; therefore, it can be used as basis for decision-making related to the number of lymph nodes that can be dissected in operations.

Modulation of α7nAchR by Melatonin Alleviates Ischemia and Reperfusion-Compromised Integrity of Blood-Brain Barrier Through Inhibiting HMGB1-Mediated Microglia Activation and CRTC1-Mediated Neuronal Loss.

The only food and drug administration (FDA)-approved drug currently available for the treatment of acute ischemic stroke is tissue plasminogen activator (tPA), yet the therapeutic benefits of this drug are partially outweighed by the increased risk of hemorrhagic transformation (HT). Analysis of the NIH trial has shown that cigarette smoking protected tPA-treated patients from HT; however, the underlying mechanism is not clear. Nicotinic acetylcholine receptors (nAChR) has shown anti-inflammatory effect and modulation nAChR could be a strategy to reduce ischemia/reperfusion-induced blood-brain barrier (BBB) damage. Since melatonin could regulate the expression of α7nAchR and melatonin's neuroprotective effect against ischemic injury is mediated via α7nAChR modulation, here, we aim to test the hypothesis that melatonin reduces ischemia and reperfusion (I/R)-induced BBB damage through modulation of α7nACh receptor (α7nAChR). Mice were subjected to 1.5 h ischemia and 24 h reperfusion and at the onset of reperfusion, mice received intraperitoneal administration (i.p.) of either drug or saline. Mice were randomly assigned into five groups: Saline; α7nAChR agonist PNU282987; Melatonin; Melatonin+Methyllycaconitine (MLA, α7nAChR antagonist), and MLA group. BBB permeability was assessed by detecting the extravasation of Evan's blue and IgG. Our results showed that I/R significantly increased BBB permeability accompanied by occludin degradation, microglia activation, and high mobility group box 1 (HMGB1) release from the neuron. In addition, I/R significantly induced neuronal loss accompanied by the decrease of CREB-regulated transcriptional coactivator 1 (CRTC1) and p-CREB expression. Melatonin treatment significantly inhibited the above changes through modulating α7nAChR. Taken together, these results demonstrate that melatonin provides a protective effect on ischemia/reperfusion-induced BBB damage, at least in part, depending on the modulation of α7nAChR.

Graphitization and Performance of Deadman Coke in a Large Dissected Blast Furnace.

The graphitization and performance of deadman coke in the blast furnace hearth have an essential influence on the longevity of the blast furnace. In this paper, coke samples were obtained from various heights in a hearth during the overhaul of the blast furnace. The voidage, particle size, graphitization degree, microstructure, and structure evolution of multiple cokes were analyzed through digital image processing, XRD, Raman spectra, scanning electron microscopy, and energy-dispersive X-ray spectroscopy (SEM-EDS). The graphitization results were compared with feed coke, tuyere coke, cohesive zone coke, and deadman coke in reference, and the main findings were analyzed. The following results were obtained. First, the voidage of deadman coke increased and then decreased with the increase of the depth while the particle size continued to decrease. In addition, the consumption rate of coke as a carburizer, reductant, and heart source was 8.47, 30.95, and 60.58%, respectively. Second, the graphitization degree of deadman coke was extremely high and showed a trend of first increasing and then decreasing. Finally, the evolution mechanism of coke graphitization was proposed. Molten iron, alkali metal, temperature, and mineral were the crucial factors that affect the graphitization of coke. The turning point of the graphitization degree was related to the buoyancy of the hearth.

Emerging roles of circUBAP2 targeting miR-370-3p in proliferation, apoptosis, and invasion of papillary thyroid cancer cells.

Circular RNAs (circRNAs) have been documented to be aberrantly expressed in many types of malignancies and involved in cancer progression. However, their role in thyroid cancer (TC) remains largely unknown. Our study aimed to explore the role and mechanism of circUBAP2 in TC. The differentially expressed circRNAs in TC tissues were identified using GSE18105 from gene expression omnibus (GEO) database. CircUBAP2 and miR-370-3p expression was analyzed using qRT-PCR. The stability of circUBAP2 was confirmed by actinomycin D and RNase R. The subcellular localization of circUBAP2 was detected using cell fractionation assay. Cell proliferation, apoptosis, and invasion were evaluated using MTT, flow cytometry analysis, and Transwell invasion assay, respectively. The interaction between circUBAP2 and miR-370-3p was predicted using bioinformatics analysis and validated by luciferase reporter assay, RNA pull-down assay, and RNA immunoprecipitation. CircUBAP2 was upregulated and miR-370-3p was downregulated in TC tissues and cells. CircUBAP2 was highly stable, resistant to RNase R digestion, and predominantly localized in the cytoplasm. CircUBAP2 knockdown inhibited cell proliferation and invasion and triggered apoptosis in TC cells. Bioinformatics analysis showed that circUBAP2 contained putative binding sites of miR-370-3p. CircUBAP2 acted as a sponge to inhibit miR-370-3p expression. Mechanistically, miR-370-3p inhibition abolished the effects of circUBAP2 on proliferation, apoptosis, and invasion in TC cells. Taken together, CircUBAP2 knockdown impeded the proliferation and invasion and induced apoptosis in TC cells via sponging miR-370-3p.

circZFR regulates thyroid cancer progression by the miR-16/MAPK1 axis.

Previous studies have identified the dysregulation of various circRNAs in many types of human cancers including thyroid cancer (TC). Circular RNA ZFR (circZFR) serves as an oncogenic circRNA in TC. However, the detailed molecular mechanism of circZFR in TC progression remains to be further explored. CircZFR and miR-16 expressions in TC cells were analyzed through qRT-PCR. Cell viability, invasion, and apoptosis were detected using CCK-8, transwell invasion assay, and flow cytometry analysis, respectively. The relationship between circZFR and miR-16 was explored using luciferase reporter assay, RNA pull-down assay, and qRT-PCR. The relationship between miR-16 and mitogen-activated protein kinase 1 (MAPK1) was explored using luciferase reporter assay and western blot analysis. Results showed that circZFR was upregulated and miR-16 was downregulated in TC cells. CircZFR knockdown inhibited the viability and invasion and induced apoptosis in TC cells. CircZFR inhibited miR-16 expression by sponging miR-16 and miR-16 repressed MAPK1 expression by targeting MAPK1. Moreover, circZFR positively regulated MAPK1 expression in TC cells by serving as a ceRNA of miR-16. Mechanistically, circZFR knockdown-induced inhibition of cell viability and invasion and promotion of apoptosis were overturned after miR-16 downregulation and promotion of MAPK1. Collectively, circZFR knockdown retarded TC progression by sponging miR-16 and modulating MAPK1 expression.

A Comprehensive Review of Characterization Methods for Metallurgical Coke Structures.

The structure of coke affects its reactivity and strength, which directly influences its performance in the blast furnace. This review divides coke structures into chemical structure, physical structure, and optical texture according to their relevant characteristics. The focuses of this review are the current characterization methods and research status of the coke structures. The chemical structures (element composition and functional group) can be characterized by elemental analysis, Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy (Raman), X-ray photoelectron spectroscopy (XPS), and nuclear magnetic resonance imaging technology (13C NMR). The physical structures (pore structure and micro-crystallite structure) can be characterized by image method, X-ray CT imaging technique, mercury intrusion method, nitrogen gas adsorption method, X-ray diffraction method (XRD), and high-resolution transmission electron microscopy (HRTEM). The optical textures are usually divided and counted by a polarizing microscope. In the end, this review provides an idea of the construction of a coke molecular structural model, based on the above characterization. With the coke model, the evolution principles of the coke can be calculated and simulated. Hence, the coke performance can be predicted and optimized.

Panax notoginseng saponins and their applications in nervous system disorders: a narrative review.

Panax notoginseng saponins (PNS), also called "sanqi" in Chinese, are the main active ingredients which are extracted from the root of Panax notoginseng (Burk.) F. H. Chen., and they have been traditionally used as a medicine in China for hundreds of years with magical medicinal value. PNS have varied biological functions, such as anti-inflammatory effects, anti-cancer effects, anti-neurotoxicity, and the prevention of diabetes. Nervous system disorders, a spectrum of diseases originating from the nervous system, have a significant impact on all aspects of patients' lives. Due to the dramatic gains in global life expectancy, the prevalence of nervous system disorders is growing gradually. Even if the mechanism of these diseases is still not clear, they are mainly characterized by neuronal dysfunction and neuronal death. Consequently, it is essential to find measures to slow down or prevent the onset of these diseases. At present, traditional Chinese medicines, as well as their active components, have gained widespread popularity in preventing and treating these diseases because of their merits, especially PNS. In this review, we predominantly address the recent advances in PNS researches and their biological functions, and highlight their applications in nervous system disorders, such as Alzheimer's disease (AD), Parkinson's disease (PD), and stroke.

GABAergic Inhibition of Spinal Cord Dorsal Horns Contributes to Analgesic Effect of Electroacupuncture in Incisional Neck Pain Rats.

BACKGROUND: Acupuncture has shown to be effective in relieving post-surgical pain. Nonetheless, its underlying mechanisms remain largely unknown. In the present study, we investigated the effect of electroacupuncture (EA) on the expression of GABA, GABA-A receptor (R) and GABA-BR in the spinal cord dorsal horns (DHs), and the involved neural cells in rats with incisional neck pain. MATERIALS AND METHODS: Male SD rats were randomly divided into control, model, Futu (LI18), Hegu-Neiguan (LI4-PC6), and Zusanli-Yanglingquan (ST36-GB34) groups. The incisional neck pain model was established by making a longitudinal incision and repeated mechanical separation along the thyroid gland region. EA (2Hz/100Hz, 1mA) was applied to LI18, LI4-PC6, ST36-GB34 separately for 30min, once at 4, 24 and 48h after incision. The local thermal pain threshold (TPT) of the focus was measured and the expression of GABA, and GABAR proteins and mRNAs detected by immunofluorescence stain and quantitative RT-PCR, respectively. RESULTS: The analgesic effect of LI18 and LI4-PC6 was superior to that of ST36-GB34 in incisional neck pain rats. Moreover, the EA stimulation of LI18 or LI4-PC6 increased the expression of GABA and GABA-Aα2 and GABA-Aß3, GABA-B1, and GABA-B2 mRNAs in spinal DHs 4h after surgery, while GABA-A and GABA-B antagonists inhibited the analgesic effect of LI18. Immunofluorescence double staining showed that GABA was expressed on astrocytes and neurons, and GABA-B expressed only on neurons. CONCLUSION: EA of both LI18 and LI4-PC6 has a good analgesic effect in incisional neck pain rats, which is closely related to their effects in upregulating the expression of GABA and its receptors in spinal DHs. The effects of LI18 and LI4-PC6 EA are obviously better that those of ST36-GB34 EA, and GABA is expressed on neurons and astrocytes.

Characteristics of novel Ti-10Mo-xCu alloy by powder metallurgy for potential biomedical implant applications.

When biomaterials are implanted in the human body, the surfaces of the implants become favorable sites for microbial adhesion and biofilm formation, causing peri-implant infection which frequently results in the failure of prosthetics and revision surgery. Ti-Mo alloy is one of the commonly used implant materials for load-bearing bone replacement, and the prevention of infection of Ti-Mo implants is therefore crucial. In this study, bacterial inhibitory copper (Cu) was added to Ti-Mo matrix to develop a novel Ti-Mo-Cu alloy with bacterial inhibitory property. The effects of Cu content on microstructure, tensile properties, cytocompatibility, and bacterial inhibitory ability of Ti-Mo-Cu alloy were systematically investigated. Results revealed that Ti-10Mo-1Cu alloy consisted of α and ß phases, while there were a few Ti2Cu intermetallic compounds existed for Ti-10Mo-3Cu and Ti-10Mo-5Cu alloys, in addition to α and ß phases. The tensile strength of Ti-10Mo-xCu alloy increased with Cu content while elongation decreased. Ti-10Mo-3Cu alloy exhibited an optimal tensile strength of 1098.1 MPa and elongation of 5.2%. Cytocompatibility study indicated that none of the Ti-10Mo-xCu alloys had a negative effect on MC3T3-E1 cell proliferation. Bacterial inhibitory rates against S. aureus and E. coli increased with the increase in Cu content of Ti-10Mo-xCu alloy, within the ranges of 20-60% and 15-50%, respectively. Taken together, this study suggests that Ti-10Mo-3Cu alloy with high strength, acceptable elongation, excellent cytocompatibility, and the bacterial inhibitory property is a promising candidate for biomedical implant applications.

Specific Knockdown of α-Synuclein by Peptide-Directed Proteasome Degradation Rescued Its Associated Neurotoxicity.

α-Synuclein (α-syn) overload is strongly associated with Parkinson disease (PD), and reduction of the α-syn level by targeting the peptide-based system through the autophagy-lysosomal pathway (ALP) is a promising strategy to delay PD progression. However, if the ALP is comprised, targeting the peptide-based proteasomal degradation system would be a good alternative. In this study, we designed a fusion peptide containing an α-syn-binding domain and a short strong proteasome-targeting motif. Our results reveal that this peptide could specifically bind to α-syn, and direct it to the proteasomes for degradation in a recombinant expression system. Furthermore, by adding a membrane-penetrating motif to this fusion peptide, we demonstrated that it could penetrate into cells and consequently suppress the cellular α-syn level through proteasome degradation in a dose- and time-dependent manner. Functionally, these effects rescued the mitochondrial dysfunction and cellular defects caused by α-syn overexpression in the cultured cells and primary neurons.

Glucose Drives Growth Factor-Independent Esophageal Cancer Proliferation via Phosphohistidine-Focal Adhesion Kinase Signaling.

BACKGROUND & AIMS: Most targeted therapies against cancer are designed to block growth factor-stimulated oncogenic growth. However, response rates are low, and resistance to therapy is high. One mechanism might relate to the ability of tumor cells to induce growth factor-independent proliferation (GFIP). This project aims to understand how (1) cancer cells preferentially derive a major growth advantage by using critical metabolic products of glucose, such as phosphoenolpyruvate (PEP), to drive proliferation and (2) esophageal squamous cell carcinoma (ESCC) cells, but not esophageal adenocarcinoma cells, can induce GFIP by using glycolysis to activate phosphohistidine (poHis)-mediated signaling through focal adhesion kinase (FAK). METHODS: The hypothesis to be tested is that ESCC GFIP induced by glucose is facilitated by PEP-mediated histidine phosphorylation (poHis) of FAK, leading to the possibility that ESCC progression can be targeted by blocking poHis signaling. Biochemical, molecular biological, and in vivo experiments including bromodeoxyuridine/5-ethynyl-2'-deoxyuridine labeling, radioisotope tracing, CRISPR gene editing, and analysis of signaling gene sets in human cancer tissues and xenograft models were performed to define the mechanisms underlying ESCC GFIP. RESULTS: Glucose promotes growth factor-independent DNA replication and accumulation of PEP in ESCC cells. PEP is the direct phospho-donor to poHis58-FAK within a known "HG" motif for histidine phosphorylation. Glucose-induced poHis58 promotes growth factor-independent FAK-mediated proliferation. Furthermore, glucose activates phosphatidylinositol-3'-kinase/AKT via poHis58-FAK signaling. Non-phosphorylatable His58A-FAK reduces xenograft growth. CONCLUSIONS: Glucose induces ESCC, but not esophageal adenocarcinoma GFIP via PEP-His58-FAK-AKT signaling. ESCC progression is controlled by actionable growth factor-independent, glucose-induced pathways that regulate proliferation through novel histidine phosphorylation of FAK.

Endothelial nitric oxide synthase enhancer AVE3085 reverses endothelial dysfunction induced by homocysteine in human internal mammary arteries.

Homocysteine (Hcy) is an independent risk factor for endothelial dysfunction in cardiovascular diseases. We hypothesized that the eNOS transcription enhancer AVE3085 may protect the endothelial function damaged by Hcy in the human internal mammary artery (IMA). Cumulative concentration-relaxation curves to acetylcholine (-10 to -4.5 log mol/L) or sodium nitroprusside were established in IMA from patients undergoing coronary artery surgery precontracted by U46619 (-8 log mol/L) in the absence/presence of Hcy (100 µmol/L) with/without AVE3085 (30 µmol/L) in vitro in a myograph. RT-qPCR and ELISA were used to quantify the mRNA and protein levels of eNOS. Colorimetric assay method was used to detect the production of nitric oxide (NO). Maximal relaxation was significantly attenuated by Hcy in human IMA. Co-incubation with AVE3085 protected endothelium from the impairment by Hcy and increased the production of NO. Exposure to Hcy for 24 h downregulated eNOS protein expression (P < 0.05) whereas it upregulated the expression of eNOS at mRNA levels (P < 0.05). The presence of AVE3085 in addition to Hcy significantly increased the eNOS protein (P < 0.05) and slightly decreased the mRNA level. The study for the first time revealed that in the human blood vessels (IMA) the clinically-relevant high concentration of Hcy directly causes endothelial dysfunction by downregulating eNOS protein that may be reversed by AVE3085. These findings not only provide new direction for protecting endothelium during coronary artery bypass grafting and improving long-term patency of the grafts, but also provide evidence to the use of eNOS enhancer in the patients with endothelial dysfunction in various pathological conditions.

Promoter methylation of WNT inhibitory factor-1 may be associated with the pathogenesis of multiple human tumors.

AIM: We investigated the association of WNT inhibitory factor-1 (WIF-1) gene methylation with the pathogenesis of multiple human tumors, using a meta-analysis based approach. MATERIALS AND METHODS: Electronic databases and manual search was additionally employed to retrieve relevant published literature. The cohort studies relating to tumor and WIF-1 were screened based on predefined selection criteria, and all extracted data from the selected studies were analyzed through STATA software. RESULTS: Sixteen studies were finally enrolled in our study involved 1112 tumor samples and 612 adjacent normal samples. The study result showed that WIF-1 gene methylations in tumor tissues were significantly higher compared with adjacent/normal tissues. The result of subgroup analysis on ethnicity revealed that in the Caucasians, Asians, and Africans, the methylation status of WIF-1 gene in tumor tissues was higher than adjacent/normal tissues. Further subgroup analysis on disease types revealed that WIF-1 gene methylation status is a widespread phenomenon that is, observed in tumor tissues of patients with multiple human tumors compared with that in adjacent/normal tissues. Interestingly, there was no significant difference in WIF-1 gene methylation between tumor tissues among patients with lung cancer, gastric cancer, astrocytoma, and adjacent/normal tissues, indicating the WIF-1 gene methylation not a general nonspecific phenomenon. CONCLUSION: WIF-1 gene methylation in tumor tissues was significantly more frequent as compared to that in adjacent normal tissues, indicating that WIF-1 gene methylation may be an important event in the pathogenesis of multiple human tumors.