Prenatal High-Sucrose Diet Induced Vascular Dysfunction of Renal Interlobar Arteries in the Offspring via PPARγ-RXRg-ROS/Akt Signaling.

SCOPE: Prenatal nutrition imbalance correlates with developmental origin of cardiovascular diseases; however whether maternal high-sucrose diet (HS) during pregnancy causes vascular damage in renal interlobar arteries (RIA) from offspring still keeps unclear. METHODS AND RESULTS: Pregnant rats are fed with normal drinking water or 20% high-sucrose solution during the whole gestational period. Swollen mitochondria and distributed myofilaments are observed in vascular smooth muscle cells of RIA exposed to prenatal HS. Maternal HS increases phenylephrine (PE)-induced vasoconstriction in the RIA from adult offspring. NG-Nitro-l-arginine (L-Name) causes obvious vascular tension in response to PE in offspring from control group, not in HS. RNA-Seq of RIA is performed to reveal that the gene retinoid X receptor g (RXRg) is significantly decreased in the HS group, which could affect vascular function via interacting with PPARγ pathway. By preincubation of RIA with apocynin (NADPH inhibitor) or capivasertib (Akt inhibitor), the results indicate that ROS and Akt are the vital important factors to affect the vascular function of RIA exposure to prenatal HS. CONCLUSION: Maternal HS during the pregnancy increases PE-mediated vasoconstriction of RIA from adult offspring, which is mainly related to the enhanced Akt and ROS regulated by the weakened PPARγ-RXRg.

Case Report: TRPV4 gene mutation causing neuronopathy, distal hereditary motor, type VIII.

Neuronopathy, distal hereditary motor, type VIII is an exceedingly rare autosomal dominant genetic disorder, also known as congenital non-progressive distal spinal muscular atrophy. It is characterized by progressive weakness in distal motor function and atrophy of muscles, without accompanying sensory impairment. Presently, there is limited literature on this condition, and accurate epidemiological data regarding its incidence remains unavailable. We report a paediatric case of distal hereditary motor, type VIII that is caused by a heterozygous missense mutation in the TRPV4 gene (NM_021625): c.805C>T. The proband is a 7-year-old male child. During pregnancy, his mother had prenatal ultrasound revealing "inward turning of the feet", a condition persisting after birth. The proband is currently unable to stand independently, exhibiting bilateral clubfoot deformity. Although possessing normal cognitive function, he cannot walk unaided. Computed radiography findings reveal pelvic tilt, bilateral knee joint valgus, and bilateral clubfoot. The patient underwent familial exome sequencing, revealing a mutation in the TRPV4 gene (NM_021625): c.805C>T (p.Arg269Cys). Considering the patient's medical history, clinical manifestations, imaging studies, and genetic test results, the diagnosis for this individual is Neuronopathy, distal hereditary motor, type VIII. This report documents a case involving the TRPV4 gene mutation associated with Neuronopathy, distal hereditary motor, type VIII, contributing valuable case reference for the early diagnosis of this condition.

Practice guideline on ovarian tissue cryopreservation and transplantation in the prevention and treatment of iatrogenic premature ovarian insufficiency.

Premature ovarian insufficiency (POI) refers to the decline of ovarian function before the age of 40. POI causes a reduction in or loss of female fertility, accompanied by different degrees of menopausal symptoms, which increases the risk of chronic diseases related to early menopause and seriously affects patients' quality of life and health. It is conservatively estimated that at least one million prepubertal girls and women of reproductive age in China are at risk of iatrogenic POI caused by radiotherapy and chemotherapy every year. With the development of medical technology and the breakthrough of scientific and technological advances, preventing and treating iatrogenic POI have become possible. International and national guidelines consider cryopreserved ovarian tissue transplantation to be the most promising method of preserving the ovarian function and fertility of prepubertal girls and women of reproductive age who cannot delay radiotherapy and chemotherapy. In order to guide the clinical application of ovarian tissue cryopreservation and transplantation technology in China, the Guideline Working Group finally included 14 scientific questions and 18 recommendations through a questionnaire survey, field investigation, and consultation of a large number of Chinese and English literature databases in order to provide a reference for colleagues in clinical practice.

Migrasome, a novel organelle, differs from exosomes.

Migrasomes, a newly discovered organelle produced by migrating cells, are vesicles with membranous structure that form on the tips and intersections of retraction fibers (RFs). These structures are released into the extracellular environment or taken up by surrounding cells, mediating the release of cytoplasmic contents and intercellular communication. Retractosomes, a new type of small extracellular vesicles generated from broken-off RFs, are closely related to migrasomes in their physical location and origin, but were defined later. Despite their widespread existence in cells and biological organisms, little is known about the regulatory mechanisms underlying their formation and potential function. In this review, we provide an overview of the discovery, biogenesis, distribution, and functions of migrasomes and retractosomes, as well as their differences from exosomes.

A facile strategy to prepare FeNx decorated PtFe intermetallic with excellent acidic oxygen reduction reaction activity and stability.

The construction of low-Pt-content intermetallic on carbon supports has been verified as a promising method to promote the activity of the oxygen reduction reaction (ORR). In this study, we have developed a simple and effective strategy to obtain a well-designed CNT-PtFe-PPy precursor. This precursor contains modulated Pt- and Fe-based content dispersed in polypyrrole (PPy) chain segments, which are in-situ generated on the templates of carbon nanotubes (CNTs). Subsequent pyrolysis of the CNT-PtFe-PPy precursor produces a CNT-PtFe@FeNC catalyst, which contains both Fe-Nx and PtFe intermetallic active sites. Due to the highly efficient dispersion of active species, the CNT-PtFe@FeNC electrocatalyst displays a 9.5 times higher specific activity (SA) and 8.5 times higher mass activity (MA) than those of a commercial Pt/C catalyst in a 0.1 M HClO4 solution. Additionally, these results, combined with excellent durability (the SA and MA maintained 94 % and 91 % of initial activity after a 10-k cycle accelerated durability test), represent among the best performance achieved so far for Pt-based ORR electrocatalysts. Furthermore, density functional theory (DFT) calculations revealed that the presence of Fe-N4 species reduces the adsorption energy between the PtFe intermetallic compound and OH*, accelerating the ORR process.

In vitro antifungal and antibiofilm activities of auranofin against itraconazole-resistant Aspergillus fumigatus.

BACKGROUND: Infections caused by azole-resistant Aspergillus are a rising public health threat with high mortality rates, high treatment costs and limited available antifungals, indicating an urgent need for new antifungals or strategies. Our aim was to investigate antifungal and antibiofilm activities of auranofin, an FDA-approved anti-antirheumatic drug. METHODS: Fungal susceptibility testing for auranofin was carried out by the broth-based microdilution methods. Cell viability treated by auranofin was tested by resazurin dye testing. The synergistic effect of auranofin and antifungal drugs was evaluated using checkboard assay. The inhibitory of biofilms were measured by crystal violet staining. Gene expression level analysis and enzyme activity was investigated with qRT-PCR analysis and DTNB assay. The key amino acid residues in the binding of auranofin with A. fumigatus thioredoxin reductase (AfTrxR) were indicated by structural analyses, site-directed mutagenesis, and microscale thermophoresis (MST) assays. RESULTS: Auranofin has fungicidal activity and in vitro antifungal spectrum including Aspergillus flavus, Aspergillus fumigatus, Aspergillus terreus, Aspergillus niger, even itraconazole (ITC)-resistant A. fumigatus. Additionally, it has antibiofilm activities against ITC-resistant A. fumigatus by reducing the expression level of SomA and MedA. Moreover, we discovered a synergistic effect of auranofin and ITC or amphotericin B against ITC-resistant A. fumigatus. Auranofin downregulated the gene transcription of AfTrxR, and strongly inhibited the enzyme activity of AfTrxR through interacting with residues C145 and C148. CONCLUSIONS: Auranofin has fungicidal and antibiofilm activities in Aspergillus spp. and is also a potentiator of ITC or amphotericin B in vitro.

Effect of reservoir characteristics and chemicals on filtration property of water-based drilling fluid in unconventional reservoir and mechanism disclosure.

The research objective of this investigation is to explore the influence of filtrate reducer and reservoir characteristics on the filtration reduction of drilling fluid during the drilling process, and the filtration reduction mechanism of drilling fluids is also revealed. The results obtained that a synthetic filtrate reducer can significantly reduce the filtration coefficient than that of the commercial filtrate reducer. Moreover, the filtration coefficient of drilling fluid constructed from synthetic filtrate reducer is reduced from 4.9 × 10-2 m3·min1/2 to 2.4 × 10-2 m3·min1/2 with an increase in the filtrate reducer content, which is much lower than that of the commercial filtrate reducer. The weaker filtration capacity of the drilling fluid containing the modified filtrate reducer is attributed to the combined action of the filtrate reducer containing multifunctional groups adsorbed on the sand surface and the hydration membrane adsorbed on the sand surface. Furthermore, the increase in reservoir temperature and shear rate increases the filtration coefficient of drilling fluid, indicating that low temperature and shear rate are conducive to improve the filtration capacity. Thus, the type and content of filtrate reducer are preferred during drilling in oilfield reservoir, but increasing reservoir temperature and shear rate are not recommended. It is necessary to confect the drilling mud with appropriate filtrate reducer such as the chemicals prepared herein during drilling operation.

Low-loading Pt nanoparticles combined with the atomically dispersed FeN4 sites supported by FeSA-N-C for improved activity and stability towards oxygen reduction reaction/hydrogen evolution reaction in acid and alkaline media.

Reducing the loading of Pt precious metal is the promising pathway to positively promote the large-scale application for fuel cells and water electrolysis. In this work, a composite bifunctional electrocatalyst (named Pt@FeSA-N-C) consisting of the atomically dispersed FeN4 active sites and Pt nanoparticles (NPs) is successfully prepared for oxygen reduction reaction (ORR) and hydrogen evolution reactions (HER). In the process of synthesizing precursor of Pt(OH)4-Fe-Ppy@CNFs, the Fe-Ppy@CNFs was firstly prepared where the highly dispersed Fe3+ ions were pre-anchored into polypyrrole (PPy) matrixes through in-situ polymerization on the surface of cellulose nanofibers (CNFs) and then Pt(OH)4 nano-particles were deposited on Fe-Ppy@CNFs through adjusting the pH of the solution by urea hydrolysis to obtain the Pt(OH)4-Fe-Ppy@CNFs. Compared with the commercial 20 wt.% Pt/C, the obtained Pt@FeSA-N-C possesses 5.5 wt.% low Pt loading. The strong synergistic effect of dual active sites between Pt NPs and FeN4 on one-dimensional (1D) FeSA-N-C support with a large surface area ensures effectively exposure of Fe and especial Pt active sites in the Pt@FeSA-N-C. Both ORR and HER activities of the Pt@FeSA-N-C were greatly improved in acid and alkaline media, even outperforming the commercial 20 wt.% Pt/C. Furthermore, the Pt@FeSA-N-C shows an unordinary stability, with no obvious decrease in the current density after 5000 and 1000 cycles of accelerated durability tests (ADTs) for ORR and HER processes, respectively. This work highlights a preparation strategy for the synergistic effect between low-loading Pt precious metal and non-precious metals in electrocatalytic system.

Influence of organoboron cross-linker and reservoir characteristics on filtration and reservoir residual of guar gum fracturing fluid in low-permeability shale gas reservoirs.

To effectively reduce the filtration rate of water-based fracturing fluid and promote the pressure holding effect of fracturing fluid in underground unconventional reservoirs, an efficient and clean organic-boron cross-linker was synthesized with boric acid and low alcohols. The results obtained that the synthesized organoboron cross-linker exhibits better fluid loss performance to water-based fracturing fluid than the commercially available cross-linker. This organoboron cross-linker allowed decreasing filtration coefficient more than 0.74 × 10-2 m3·min1/2 as a result of the network structure formed by the organoboron cross-linker and guar gum molecule. However, commercially available cross-linker exhibits a relatively large filtered mass of water more than 1.33 × 10-2 m3·min1/2 at the same condition. Meanwhile, the cross-linked guar gum fracturing fluid can significantly improve the fluid loss property with the increase of cross-linker content and pressure, and an increased fluid filtration gradually was revealed with increasing the reservoir temperature and current speed. Moreover, the damage of shale reservoir caused by the prepared boron cross-linker was only 11%, which was lower than 18% of the commercial boron cross-linker under the same conditions.

[Water intake and its influence factors among pregnant and lactating women in some areas of China in 2020].

OBJECTIVE: To investigate the water intake and its influence factors of pregnant and lactating women. METHODS: From May to August 2020, a convenience sampling method was used to investigate questionnaires in the obstetrics and pediatric care departments of maternal and Child Health Hospitals in Beijing, Tianjin, Shandong and Jiangsu Provinces(Cities) among pregnant and lactating women who received prenatal health care and postpartum follow-up. The questionnaire included general conditions, physical activity and water-related surveys. The data was exported from the Questionnaire Star System and analyzed by Wilcoxon rank sum test and multiple linear regression for different types of daily fluid intake. RESULTS: The median daily water intake of pregnant and lactating women was 1321 mL and 1271 mL, respectively, meanwhile, plain water was the most highest(both 1000 mL), followed by milk and milk products(179 mL and 86 mL), other beverages(29 mL and 86 mL). The one-way analysis showed that daily water intake of pregnant women was increased with increasing gestation, literacy, and household disposable income, and the difference were statistically significant(P<0.05) across gestation, literacy, physical activity intensity, and household disposable income groups, as well as significant differences in lactating women(P<0.05). Multiple linear regression analysis showed that pregnancy was the most influential factor for daily water intake, plain water and liquid milk and yogurt intake among pregnant women, while household disposable income was the most influential factor for other beverages. For lactating women, household disposable income was the most influential factor for daily water intake, liquid milk and yogurt and other beverages, and literacy was the most influential factor for daily intake of plain water. CONCLUSION: Pregnant and lactating women had insufficient water intake, so it is necessary to consider different pregnancy status, family income and literacy for drinking water health education and improve drinking behavior.

Experimental investigation on the high-pressure sand suspension and adsorption capacity of guar gum fracturing fluid in low-permeability shale reservoirs: factor analysis and mechanism disclosure.

Guar fracturing technology has been considered as a kind of popular EOR technology, but the weak static suspension capacity becomes a challenge due to the poor temperature resistance and stability of guar fracturing fluid. The main goal of this investigation is to explore the effect of different factors on the high-pressure static sand suspension of guar gum fracturing fluid by a synthetic efficient nano-ZrO2 cross-linker. In particular, a mechanism of static suspended sand of nano-ZrO2 cross-linker is analyzed by microscopic simulation. The adsorption performance of guar fracturing fluid on the shale surface is also studied for analyzing the environmental pollution and damage of guar gum fracturing fluid to shale reservoirs after cross-linking in this investigation. The results obtained that the inclusion of a small content of nano-ZrO2 cross-linker (0.4%) leads to an apparent increase of fracturing fluid viscosity and decrease in the falling quality of gravel (104 mPa·s and 0.3 g) compared to the classical cross-linker (63 mPa·s and 3.5 g). The lower adsorption capacity of guar fracturing fluid containing nano-ZrO2 cross-linker on the shale surface means that it has a weaker pollution ability to the shale reservoir than the commercially available cross-linker. Meanwhile, the grid structure density formed by nano-cross-linker and guar gum is considered to be the key factor to significantly change the suspended sand capacity. The investigation of nano-cross-linker cannot only provide necessary theoretical technology and data support for the stability of water-based fracturing fluid, efficient sand carrying, and the development of water-based fracturing technology, but also effectively protect the underground shale reservoir.

Affecting analysis of the rheological characteristic and reservoir damage of CO2 fracturing fluid in low permeability shale reservoir.

The fracturing property of liquid CO2 fracturing fluid varies greatly due to the rheology of fracturing fluid during fracturing process. The main objective of this investigation is to study the rheology property of thickened liquid CO2 by measuring the viscosity of thickened liquid CO2 in different physical parameters of this prepared thickener and explain the causes of rheological changes. The results show that thickener content, branching content, and molecular weight of a thickener for all could significantly improve the rheology of liquid CO2; the consistency coefficient K increased as they rose, but the rheological index n presented a decreased trend. Meanwhile, the mesh structure is proposed as a model to explain the rheological changes, and the large wetting angle means an excellent backflow, low reservoir damage, and low adsorption property. These results herein provide a basic reference to improve the CO2 fracturing technology and molecular design of CO2 thickener.

Factor analysis and mechanism disclosure of supercritical CO2 filtration behavior in tight shale reservoirs.

As an important working fluid in tight shale reservoir, supercritical CO2 has been proven to improve oil recovery efficiently. However, the high filtration caused by the low viscosity of pure supercritical CO2 hinders its development. The research objective of this investigation is to explore the filtration of supercritical CO2 with a branched siloxane (BTMT) as a CO2 thickener and filtration-reducing agent, and analyze the influence level of some parameters about rock core and chemicals on the CO2 filtration in the tight shale reservoir by using response surface method (RSM). The results demonstrate that the rising temperature causes a gradually increasing filtration, but filtration coefficient (f) decreases with increasing the pressure difference P, injection speed, and thickener concentration. The thickener concentration is the factor that causes the greatest change in filtration coefficient according to the response surface method, and the injection speed has the smallest effect on the filtration. The viscosity of fracturing fluid is the main characterization parameter leading to change of filtration coefficient, all factors that contribute to increasing the viscosity of the fracturing fluid will lead to a reduction in the filtration coefficient and an enhanced oil recovery. In addition, the adsorption and reservoir residue of BTMT on low-permeability shale were subordinated to a Langmuir monolayer theory, and a low residual of BTMT in shale can prevent thickeners and fracturing fluids from damaging shale reservoirs. The improvement of thickener and CO2 fracturing technology provided a basic reference for shale exploitation, greenhouse effect, and reservoir protection.

Conversion of rice husk biomass into electrocatalyst for oxygen reduction reaction in Zn-air battery: Effect of self-doped Si on performance.

An outstanding oxygen reduction reaction (ORR) electrocatalyst is firstly developed deriving from sustainable rice husk (RH) biomass. Benefiting from self-doped Si in RH, the higher proportion of pyridine N, graphite N and expecially Fe-Nx as well as thiophene S contents were produced in Si-Fe/S/N-RH3 in comparison with those of Si-free Fe/S/N-RH3. Consequently, the half-wave potential of 0.89 V and the onset potential of 0.96 V are achieved for Si-Fe/S/N-RH3, outperforming the benchmark electrocatalyst Pt/C and other Fe-based electrocatalysts reported in alkaline media. Furthermore, it is found that the exisentence of self-doped Si can improve the graphitization degree of the catalyst, leading to the long-term stability (larger than 85% retention after 40000 s) and prominent methanol tolerance for Si-Fe/S/N-RH3. In addition, Si-Fe/S/N-RH3 shows a power density of 86.2 mW cm-2 and excellent durability in Zn-air battery. The work highlights the potential to develop sustainable and cost-effective ORR electrocatalysts from waste biomass as the substitute for precious metal catalysts.

Optimization the extraction of anthocyanins from blueberry residue by dual-aqueous phase method and cell damage protection study.

Blueberry residue is usually discarded as waste, but has a high anthocyanins content. The extraction method of anthocyanins from blueberry residue with ultrasonic assisted dual-aqueous phase system was optimized. In terms of the principle of central group and design (CCD) experimental design, three-factor and five-level response surface analysis was adopted to optimize the extraction conditions with the extraction rate of anthocyanins. The optimum extraction rate of anthocyanin was 12.372 ± 0.078 mg/g. Anthocyanin extract could protect the pBR322 DNA oxidative damage induced by Fenton reagent, increase the superoxide dismutase(SOD) and glutathione peroxidase (GSH-Px) enzyme activities, and decrease the H2O2-induced cell apoptosis of human normal liver cell (LO2 cell). The study indicates that the extraction rate of anthocyanin was increased by optimized ultrasonic assisted dual-aqueous phase system. The anthocyanin extract could protect DNA and LO2 cell from oxidative damage.

B3GNT3 acts as a carcinogenic factor in endometrial cancer via facilitating cell growth, invasion and migration through regulating RhoA/RAC1 pathway-associated markers.

BACKGROUND: Aberrant expression of beta-1,3-N-acetylglucosaminyltransferase-3 (B3GNT3) has been frequently clarified in various cancers, however, its role in endometrial cancer (EC) has not been assessed in detail. PURPOSE: This study aimed to investigate the biological role of B3GNT3 in EC and simply explored the detailed mechanism. METHODS: The EC RNA-Seq dataset from TCGA database was applied to evaluate the expression of B3GNT3 and assess its role on prognostic value. HEC-1-A and KLE cell lines of EC were used to perform loss- and gain-of-function B3GNT3 assays respectively. Quantitative real-time PCR (qRT-PCR) and western blot were used to measure the mRNA and protein levels of indicated molecules respectively. Cell counting kit-8, clone formation tests, and Transwell assay served to determine the changes of proliferative, invasive and migratory abilities of EC cells after altering the expression of B3GNT3. RESULTS: B3GNT3 was found to be highly expressed in EC tissues compared to normal tissues according to the online public databases, which confirmed by the following qRT-PCR in 3 EC cell lines. Besides, high B3GNT3 expression presented a worse overall survival in EC patients as compared with low B3GNT3 expression group. Furthermore, functional experiments in vitro indicated that B3GNT3 could facilitate the cell growth, invasion and migration. Moreover, we found that downregulation of B3GNT3 significantly reduced the expression level of GTP-RhoA and GTP-RAC1, whereas upregulation of B3GNT3 presented the opposite results. CONCLUSION: The results of current study demonstrate that B3GNT3 acts as an oncogene that promotes EC cells growth, invasion and migration possibly through regulating the RhoA/RAC1 signaling pathway-related markers, suggesting that B3GNT3 may be a candidate biomarker for EC therapeutic intervention.

Endophytic Fungi from Dalbergia odorifera T. Chen Producing Naringenin Inhibit the Growth of Staphylococcus aureus by Interfering with Cell Membrane, DNA, and Protein.

This study focused on the antibacterial effects of the endophytic fungi producing naringenin from Dalbergia odorifera T. Chen against Staphylococcus aureus. The antibacterial activity was measured by the inhibition diameters, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC). The time-killing curve was also used to evaluate its antibacterial efficacy. The results of antibacterial activity determinations showed that endophytic fungi secondary metabolites can inhibit the growth of five pathogenic bacteria (S. aureus, Escherichia coli, Salmonella enteritidis, Pseudomonas aeruginosa, and Bacillus subtilis) and the most sensitive strain was S. aureus that had the MIC and MBC values of 0.13 and 0.50 mg/mL, respectively. The membrane permeability study was measured by a DNA leakage assay and electrical conductivity assay. Furthermore, the whole-cell protein lysates and DNA fragmentation assay was evaluated. The morphology of S. aureus treated with the endophytic fungi products was observed by scanning electron microscopy (SEM). The probable antibacterial mechanism of endophytic fungi secondary metabolites was the increased membrane permeability that leads to leaks of nucleic acids and proteins. SEM results further confirmed that the extracts can interfere with the integrity of S. aureus cell membrane and further inhibit the growth of bacteria, resulting in the death of bacteria. This study provides a new perspective for the antibacterial functions of endophytic fungi secondary metabolites for biomedical applications.

Optimization of trypsin extraction technology of Allium cepa L. polysaccharide by response surface methodology and the antitumor effects through immunomodulation.

The trypsin-assisted extraction of polysaccharides from Allium cepa L. was optimized using the response surface methodology (RSM). The optimum extraction conditions were extraction temperature, extraction time, extraction pH, and enzyme amount of 37.16°C, 180 min, 8.57, and 5.16%, respectively. Under the optimized conditions, the yield of A. cepa L. polysaccharides (ACP) reached 9.69%, which was comparable with the predicted yield (9.73%). Mid- and high-dose ACP significantly inhibited the tumor growth (43.93%) and the tumor inhibition percentage (38.05%), which were more than 30%. The ACP could extend the survival time of H22 ascites tumor-bearing mice. Furthermore, the ACP could reduce the thymus and the spleen atrophy and significantly promoted the Con A-induced proliferation of splenocytes and elevated the serum IFN-γ and IL-2 levels. Therefore, the ACP could inhibit the tumor growth in tumor-bearing mice and regulated the immune function of mice. Practical ApplicationsThe trypsin-assisted extraction has high efficiency, is carried out through the polysaccharide extraction and the deproteinization at the same time, and is more convenient and fast than traditional methods. No detailed study on the optimization of the trypsin extraction of onion polysaccharides is available. Thus, this experiment aims to use the BBD (4 factors and 3 levels) to optimize the roles of extraction temperature, extraction time, extraction pH, and amount of enzyme on the yield of polysaccharides obtained from the fruit of A. cepa L. In addition, when looking for high-quality biological functional principles for the pharmaceutical industry, the antitumor activity of ACP was evaluated. A. cepa L. is one of the most widely cultivated and consumed crops worldwide. Polysaccharides are the main active ingredient, and studies have shown that a high intake of Allium vegetables is associated with reduced risk of cancers.

Baicalein suppresses growth of non-small cell lung carcinoma by targeting MAP4K3.

Exploring key genes associated with non-small cell lung carcinoma (NSCLC) may lead to targeted therapies for NSCLC patients. The protein kinase MAP4K3 has been established as an important modulator of cell growth and autophagy in mammals. Herein, we investigated the somatic mutations and the expression pattern of MAP4K3 detected in NSCLC patients based on the TCGA database. Abnormal MAP4K3 expression and its somatic mutations are associated with the carcinogenesis and thereby becoming an attractive therapeutic target. Baicalein, a natural product, was determined to be the first-reported MAP4K3 binding ligand with its KD values of 6.47 µM measured by microscale thermophoresis. Subsequent in silico docking and mutation studies demonstrated that baicalein directly binds to MAP4K3, presumably to the substrate-binding pocket of this kinase domain, causing inactivity of MAP4K3. We further showed that baicalein could induce degradation of MAP4K3 through decreasing its stability and promoting the ubiquitin proteasome pathway. Degradation of MAP4K3 could cause dissociation of the transcription factor EB and 14-3-3 complex, enhance rapid transport of TFEB to the nucleus and trigger TFEB-dependent autophagy, resulting in lung cancer cells proliferation arrest. Knockdown of MAP4K3 expression by siRNA was sufficient to mimic baicalein-induced autophagy. Ectopic expression of the MAP4K3 protein resulted in significant resistance to baicalein-induced autophagy. Baicalein exhibited good tumor growth inhibition in a nude mouse model for human H1299 xenografts, which might be tightly related to its binding to MAP4K3 and degradation of MAP4K3. Our data provide novel mechanistic insights of baicalein/ MAP4K3/ mTORC1/ TFEB axis in regulating baicalein-induced autophagy in NSCLC, suggesting potential therapies for treatment of NSCLC.

Sulfated modification, characterization and monosaccharide composition analysis of Undaria pinnatifida polysaccharides and anti-tumor activity.

Undaria pinnatifida (U. pinnatifida) polysaccharides (UPPS) are considered to be the major bioactive components of U. pinnatifida. The aim of the present study was to investigate the separation, sulfated modification, characterization and monosaccharide composition of UPPS. The optimal processing conditions were as follows: Distilled water-to-solid ratio, 50 ml/g; extraction time, 300 min; and extraction temperature, 90˚C. The major polysaccharide fraction of U. pinnatifida (UPPS-B1) was purified via DEAE-52 and Sephadex G-200 column chromatography. The chlorosulfonic acid-pyridine method was applied for sulfation modification. UPPS-B1 and sulfated (S)-UPPS-B1 were characterized via chemical analysis, ultraviolet-visible and Fourier-transformed infrared spectroscopy, gas chromatography and high-performance liquid chromatography. The total sugar content of UPPS-B1 and S-UPPS-B1 was 79.78 and 77.28%, respectively. The sulfate radical content of UPPS-B1 and S-UPPS-B1 was 8.53 and 29.12%, whilst the content of uronic acid was 9.29 and 7.98%, respectively. The average molecular weight of UPPS-B1 and S-UPPS-B1 was determined to be 37 and 110 kD, respectively. UPPS-B1 was considered to be a heteropolysaccharide composed of xylose, mannose, glucose and galactose at a ratio of 7.9:8.7:12.0:9.8. In addition, S-UPPS-B1 was a heteropolysaccharide composed of xylose, mannose, glucose and galactose at a ratio of 1.0:9.7:6.4:1.6. The results of the tumor growth inhibition experiment demonstrated that UPPS-B1 exhibited anti-tumor activity in vivo, which was improved following sulfation to yield S-UPPS-B1.