Melatonin alleviates intrarenal CaOx crystals deposition through inhibiting LPS-induced non-canonical inflammasome-mediated renal tubular epithelial cells pyroptosis.

Urinary tract infection has long been considered a complication rather than etiology of calcium oxalate (CaOx) nephrolithiasis. This study aimed to explore the role of lipopolysaccharide (LPS), an important component of Gram-negative bacteria, on CaOx nephrolithiasis formation and antagonistic effect of melatonin. Male C57BL/6 mice were intraperitoneally injected with glyoxylate acid (80 mg/kg) daily for 7 days to construct CaOx nephrolithiasis model. A single dose of LPS (2.0 mg/kg) was given 2 h before the second glyoxylate acid treatment in the presence or absence of melatonin (25 mg/kg). Our results found that LPS promoted adhesion of CaOx crystals to renal tubular epithelial cells (RTECs) and intrarenal CaOx crystals deposition. Protein levels of cleaved Caspase-11, N-terminal of cleaved GSDMD (GSDMD-N), NOD-like receptor thermal protein domain associated protein 3 (NLRP3) and cleaved Caspase-1, several markers of non-classical inflammasome activation were upregulated in LPS-treated mouse kidneys and HK-2 cells. Moreover, the number of GSDMD pores was increased in LPS-treated HK-2 cell membrane. Melatonin inhibited Caspase-11 cleavage and antagonized the subsequent LPS-mediated upregulation of GSDMD-N, NLRP3 and cleaved Caspase-1 in kidney tissues and HK-2 cells. In addition, melatonin reduced membrane localization of GSDMD-N and the number of GSDMD pores in LPS-treated HK-2 cells. Accordingly, melatonin inhibited LPS-induced IL-1ß and IL-18 in mouse serum and HK-2 culture supernatant. Importantly, melatonin alleviated LPS-induced crystal-cell interactions and intrarenal CaOx crystals deposition. We provide experimental evidence that LPS promoted CaOx nephrolithiasis formation by inducing non-canonical inflammasome-mediated RTECs pyroptosis. Melatonin alleviated CaOx nephrolithiasis formation through inhibiting LPS-induced non-canonical inflammasome-mediated RTECs pyroptosis.

[Analysis of Histologic Transformation and Prognosis for Follicular Lymphoma].

OBJECTIVE: To investigate the related factors of invasive transformation and prognosis for follicular lymphoma. METHODS: A total of 168 patients with follicular lymphoma at First Affiliated Hospital of Zhengzhou University from August 2015 to January 2021 were collected, and the significance of each index in histological transformation (HT) and prognosis were analyzed. RESULTS: Pathology grade3, Ki-67 index ≥40%, ß2MG＞3 mg/L, LDH＞245 U/L, POD24 and non-invasion of bone marrow were associated with HT. Univariate analysis showed that the high risk of FLIPI-2, pathological grade 3, Ki-67≥40%, anemia, ß2MG＞3 mg/L, LDH＞245 U/L and HT had significant adverse effects on PFS; ß2MG＞3 mg/L, LDH＞245 U/L, POD24 and HT had significant adverse effects on OS. Cox multivariate analysis showed that the ß2MG >3 mg/L and HT were independent risk factors of PFS, HT was independent risk factor of OS. CONCLUSION: The pathological grade, Ki-67, ß2MG, LDH, POD24 and bone marrow invasion of FL can predict the risk of HT. Meanwhile, ß2MG >3 mg/L and HT are significantly related to poor prognosis of FL.

Characterization of Composite Film of Gelatin and Squid Pen Chitosan Obtained by High Hydrostatic Pressure.

In the present study, gelatin-based films incorporating squid pen chitosan obtained by high hydrostatic pressure (HHP chitosan) at varying proportions were prepared and their properties were compared with films containing untreated chitosan. The resulting films were characterized by analyzing the physical, morphological, mechanical and barrier properties. The addition of different ratios of HHP chitosan to the gelatin-based film yielded significant improvements in mechanical and moisture barrier properties. The reason for this might be that HHP chitosan contributed to a regular and dense microstructure of the composite films due to forming a three-dimensional network structure in gelatin-based films with enhanced intermolecular interactions. The FTIR spectra showed no new chemical bond formed by incorporating HHP chitosan into gelatin-based film. The SEM micrographs showed that the gelatin-based film fabricated with three types of chitosan had a homogeneous surface morphology, indicating good compatibility of the materials. Compared to the gelatin-based films containing untreated chitosan, films containing HHP chitosan significantly delayed oxidative deterioration in oil during storage. Therefore, the chitosan obtained by HHP treatment could have a potential application in edible gelatin-based films as packaging materials.

Formation of Porous Structures and Crystalline Phases in Poly(vinylidene fluoride) Membranes Prepared with Nonsolvent-Induced Phase Separation-Roles of Solvent Polarity.

PVDF membranes were prepared with nonsolvent-induced phase separation, using solvents with various dipole moments, including HMPA, NMP, DMAc and TEP. Both the fraction of the polar crystalline phase and the water permeability of the prepared membrane increased monotonously with an increasing solvent dipole moment. FTIR/ATR analyses were conducted at the surfaces of the cast films during membrane formation to provide information on if the solvents were present as the PVDF crystallized. The results reveal that, with HMPA, NMP or DMAc being used to dissolve PVDF, a solvent with a higher dipole moment resulted in a lower solvent removal rate from the cast film, because the viscosity of the casting solution was higher. The lower solvent removal rate allowed a higher solvent concentration on the surface of the cast film, leading to a more porous surface and longer solvent-governed crystallization. Because of its low polarity, TEP induced non-polar crystals and had a low affinity for water, accounting for the low water permeability and the low fraction of polar crystals with TEP as the solvent. The results provide insight into how the membrane structure on a molecular scale (related to the crystalline phase) and nanoscale (related to water permeability) was related to and influenced by solvent polarity and its removal rate during membrane formation.

Negligible Impact of Drought-Resistant Genetically Modified Maize on Arthropod Community Structure Observed in a 2-Year Field Investigation.

Dehydration-responsive element-binding (DREB) transcription factors regulate diverse processes during plant development. Here, a 2-year field study was conducted to assess the potential effects of DREB-genetically modified maize (GM1) on arthropod species and ecological communities. Arthropod abundance, diversity, and community composition in GM1 and its non-transformed counterpart maize variety, Chang 7-2, were compared using whole plant inspection, pitfall trap, and suction sampler methods. Based on Shannon-Wiener diversity, Simpson's diversity, Pielou's indexes, number of species, and total number of individuals, GM1 had a negligible effect on arthropod abundance and diversity. Redundancy analysis indicated that the composition of arthropod community was not associated with maize type in the three investigation methods, while it exhibited significant correlation with year and sampling time in whole plant inspection and suction sample methods, and distinctly correlated with sampling time in the pitfall trap method. Nonmetric multidimensional scaling analysis of variable factors in the three investigation methods showed that sampling time, rather than maize type or year, was closely related to the composition of arthropod community in the field. Our results provide direct evidence to support that DREB-GM maize had negligible effects on arthropods in the Jilin Province under natural conditions.

Sirt1 inhibits kidney stones formation by attenuating calcium oxalate-induced cell injury.

Cell injury is a necessary and critical event during CaOx kidney stone formation. Sirt1 exerts a number of pleiotropic effects, protecting against renal cell injury. This study aims to explore the relationship between Sirt1 and CaOx kidney stone formation and the underlying mechanism. Sirt1 expression in renal tissues or HK-2 cells was detected by Western blot, immunohistochemistry and immunofluorescence. Apoptosis in renal tissues was examined by TUNEL staining. Renal pathological changes and the crystals deposition were detected by hematoxylin-eosin and Von Kossa staining. Crystal-cell adhesion and cell injury in HK-2 cells were assessed by atomic absorption spectrometry and flow cytometry, respectively. Sirt1 expression in nephrolithiasis patients was downregulated and the level of apoptosis was increased. Further study found that Sirt1 expression was decreased in both in vivo and in vitro models. Interestingly, the levels of cell injury were elevated in vivo and in vitro models. Suppressing Sirt1 expression promoted COM-induced crystal-cell adhesion and exacerbated cell injury. In contrast, increasing the expression of Sirt1 by lentivirus transfection in vitro and resveratrol administration in vivo, alleviated crystal deposition and cell damage. Our findings suggest that Sirt1 could inhibit kidney stone formation, at least in part, through attenuating CaOx -induced cell injury.

Overexpression of ZmWRKY65 transcription factor from maize confers stress resistances in transgenic Arabidopsis.

Plant-specific WRKY transcription factors play important roles in regulating the expression of defense-responsive genes against pathogen attack. A multiple stress-responsive WRKY gene, ZmWRKY65, was identified in maize by screening salicylic acid (SA)-induced de novo transcriptomic sequences. The ZmWRKY65 protein was localized in the nucleus of mesophyll protoplasts. The analysis of the ZmWRKY65 promoter sequence indicated that it contains several stress-related transcriptional regulatory elements. Many environmental factors affecting the transcription of ZmWRKY65 gene, such as drought, salinity, high temperature and low temperature stress. Moreover, the transcription of ZmWRKY65 gene was also affected by the induction of defense related plant hormones such as SA and exogenous ABA. The results of seed germination and stomatal aperture assays indicated that transgenic Arabidopsis plants exhibit enhanced sensitivity to ABA and high concentrations of SA. Overexpression of ZmWRKY65 improved tolerance to both pathogen attack and abiotic stress in transgenic Arabidopsis plants and activated several stress-related genes such as RD29A, ERD10, and STZ as well as pathogenesis-related (PR) genes such as PR1, PR2 and PR5; these genes are involved in resistance to abiotic and biotic stresses in Arabidopsis. Together, this evidence implies that the ZmWRKY65 gene is involved in multiple stress signal transduction pathways.

Two-Stage Semi-Continuous 2-Keto-Gluconic Acid (2KGA) Production by Pseudomonas plecoglossicida JUIM01 From Rice Starch Hydrolyzate.

A two-stage semi-continuous strategy for producing 2-keto-gluconic acid (2KGA) by Pseudomonas plecoglossicida JUIM01 from rice starch hydrolyzate (RSH) has been developed. The initial glucose concentration (140 g/L) was selected for first-stage fermentation due to its highest 2KGA productivity of 7.58 g/(L⋅h), cell weight of 3.91 g/L, and residual glucose concentration of 25.00 g/L. Followed by removing 70.0% (v/v) of the first-stage broth and feeding 400.0 g/L of glucose to the second-stage fermentor, a total of 50680.0 g glucose was consumed, and 50005.20 g 2KGA was obtained with a yield of 0.9867 g/g by P. plecoglossicida JUIM01 after a 3-cycle two-stage semi-continuous fermentation. Our results indicated that the developed two-stage semi-continuous fermentation could be industrially applied due to its high 2KGA concentration, 2KGA yield and operation efficiency.

A Membrane-Bound Gluconate Dehydrogenase from 2-Keto-D-Gluconic Acid Industrial Producing Strain Pseudomonas plecoglossicida JUIM01: Purification, Characterization, and Gene Identification.

The membrane-bound gluconate dehydrogenase (mGADH) is a critical enzyme for 2-keto-D-gluconic acid (2KGA) production in Pseudomonas plecoglossicida JUIM01. The purified native flavin adenine dinucleotide-dependent mGADH (FAD-mGADH) was consisted of a gamma subunit, a flavoprotein subunit, and a cytochrome c subunit with molecular mass of ~ 27, 65, and 47 kDa, respectively. The specific activity of FAD-mGADH was determined as 90.71 U/mg at optimum pH and temperature of 6.0 and 35 °C. The Km and Vmax values of calcium D-gluconate were 0.631 mM and 0.734 mM/min. The metal ions Mg2+ and Mn2+ showed slight positive effects on FAD-mGADH activity. On the other hand, a 3868-bp-length gad gene cluster was amplified and expressed in Escherichia coli BL21(DE3). The recombinant protein showed the same molecular weight and enzyme activity as the native FAD-mGADH, which confirmed it as a FAD-mGADH encoding gene. The flavoprotein subunit and the cytochrome c subunit containing a putative FAD-binding motif and three possible heme-binding motifs concluded from alignment results of mGADHs. This study characterized the native and recombinant FAD-mGADH and would provide the basis for further genetic modification of Pseudomonas plecoglossicida JUIM01 with the intention of 2KGA productivity improvement.

Fouling-Resistant and Self-Cleaning Aliphatic Polyketone Membrane for Sustainable Oil-Water Emulsion Separation.

The cost-effective treatment of emulsified oily wastewater discharged by many industries and human societies is a great challenge. Herein, based on an aliphatic polyketone (PK) polymer with a good membrane formation ability and an intrinsic intermediate hydrophilicity, a new class of reduced PK (rPK) membranes combining an all hydrophilic and electrically neutral surface chemistry comprising ketone and hydroxyl groups, and a fibril-like morphology featuring re-entrant structure, was facilely prepared by phase separation and following fast surface reduction. The synergetic cooperation of surface chemistry and surface geometry endowed the prepared membranes with excellent superhydrophilicity, underwater superoleophobicity, and underoil superhydrophilicity, in addition to antiprotein-adhesion property. Thus, fouling-resistant and self-cleaning filtrations of challenging oil-in-water emulsions containing adhesive oil, surfactant, high salinity, and proteins were effortlessly realized with high flux (up to ∼50 000 L m-2 h-1 bar-1), slow and reversible flux decline, and low oil permeate (<20 ppm). In contrast, a commercial superhydrophilic microporous membrane made of mixed cellulose ester suffered severe fouling gradually or immediately when carrying out the emulsion filtrations due to its less than ideal surface properties. It is believed that this class of membranes with desirable superwettability, high flux, and preparation simplicity can be a potential new benchmark for high performance and large-scale oil-water separation in complex environments.

Dual Superlyophobic Aliphatic Polyketone Membranes for Highly Efficient Emulsified Oil-Water Separation: Performance and Mechanism.

Efficient treatment of difficult emulsified oil-water wastes is a global challenge. Membranes exhibiting unusual dual superlyophobicity (combined underwater superoleophobicity and underoil superhydrophobicity) are intriguing to realize high-efficiency separation of both oil-in-water and water-in-oil emulsions. For the first time, a robust polymeric membrane demonstrating dual superlyophobicity to common apolar oils was facilely fabricated via a simple one-step phase separation process using an aliphatic polyketone (PK) polymer, thanks to a conjunction of intermediate hydrophilicity and re-entrant fibril-like texture upon the prepared PK membrane. Further chemical modification to improve surface hydrophilicity slightly can enable dual superlyophobicity to both apolar and polar oils. It is found that a nonwetting composite state of oil against water or water against oil was obtainable on the membrane surfaces only when the probe liquids possess an equilibrium contact angle (θow or θwo) larger than the critical re-entrant angle of the textured surfaces (73°), which can explain the existences of dual superlyophobicity and also the nonwetting to fully wetting transitions. A simple design chart was developed to map out the operational windows of material hydrophilicity and re-entrant geometry, that is, a possible zone, to help in the rational design of similar interfacial systems from various materials. Switchable filtrations of oil-in-water and water-in-oil nanoemulsions were achieved readily with both high flux and high rejection. The simplicity and scalability of the membrane preparation process and the well-elucidated underlying mechanisms illuminate the great application potential of the PK-based superwetting membranes.

Scalable Wet Deposition of Zeolite AEI with a High Degree of Preferred Crystal Orientation.

Producing zeolite films with controlled preferred orientation on an industrial scale is a long-standing challenge. Herein we report on a scalable approach to the direct wet deposition of zeolite thin films and membranes while maintaining a high degree of control over the preferred crystal orientation. As a proof of concept, thin films comprising aluminophosphate zeolite AEI were cast on silicon wafer or porous alumina substrates. Electrical properties and separation performance of the zeolite thin films/membranes were engineered through controlling degree of preferred crystal orientation.

Purification, characterization and gene identification of a membrane-bound glucose dehydrogenase from 2-keto-d-gluconic acid industrial producing strain Pseudomonas plecoglossicida JUIM01.

The membrane-bound glucose dehydrogenase (mGDH) is a rate-limiting enzyme for the industrial production of 2-keto-d-gluconic acid (2KGA) from glucose. In this study, mGDH was firstly purified from a 2KGA industrial producing strain Pseudomonas plecoglossicida JUIM01. The purified mGDH exhibited a specific activity of 16.85 U/mg and was identified as monomeric membrane-bound PQQ-dependent dehydrogenase with a molecular mass of ~87 kDa. The Km and Vmax value of d-glucose were 0.042 mM and 14.620 µM/min, and the optimal pH and temperature were of 6.0 and 35 °C with favorable acid resistance and poor heat tolerance. Ca2+/Mg2+ showed a significantly positive effect on mGDH activity with 20% increase, whereas EDTA/EGTA had a negative influence, and Ca2+ was essential for enzyme activity. Furthermore, a 2412 bp-length gcd was amplified by genome walking technique and heterologously expressed in Escherichia coli. Bioinformatics analysis and heterologous expression further confirmed it as a mGDH encoding gene. mGDH contained binding sites of Ca2+, cofactor PQQ and polypeptide binding sites concluded from alignment results of mGDHs from different genera. This study would lay the foundation for improving 2KGA productivity through further strain modification.

Adsorption of Bovine Serum Albumin on Poly(vinylidene fluoride) Surfaces in the Presence of Ions: A Molecular Dynamics Simulation.

Adsorption of bovine serum albumin (BSA) on poly(vinylidene fluoride) (PVDF) surfaces in an aqueous environment was investigated in the presence and absence of excess ions using molecular dynamics simulations. The adsorption process involved diffusion of protein to the surface and dehydration of surface-protein interactions, followed by adsorption and denaturation. Although adsorption of BSA on PVDF surface was observed in the absence of excess ions, denaturation of BSA was not observed during the simulation (1 µs). Basic and acidic amino acids of BSA were found to be directly interacting with PVDF surface. Simulation in a 0.1 M NaCl solution showed delayed adsorption of BSA on PVDF surfaces in the presence of excess ions, with BSA not observed in close proximity to PVDF surface within 700 ns. Adsorption of Cl- on PVDF surface increased its negative charge, which repelled negatively charged BSA, thereby delaying the adsorption process. These results will be helpful for understanding membrane fouling phenomena in polymeric membranes, and fundamental advancements in these areas will lead to a new generation of membrane materials with improved antifouling properties and reduced energy demands.

Synthesis and Characterization of Alkali Metal Ion-Binding Copolymers Bearing Dibenzo-24-crown-8 Ether Moieties.

Dibenzo-24-crown-8 (DB24C8)-bearing copolymers were synthesized by radical copolymerization using a DB24C8-carrying acrylamide derivative and N-isopropylacrylamide monomers. The cloud point of the resulting copolymers changed in aqueous solution in the presence of cesium ions. In addition, the ¹H NMR signals of DB24C8-bearing copolymers shifted in the presence of alkali metal. This shift was more pronounced following the addition of Cs⁺ compared to Rb⁺, K⁺, Na⁺, and Li⁺ ions due to recognition of the Cs⁺ ion by DB24C8.

Extracranial Internal Carotid Artery Tortuosity and Body Mass Index.

BACKGROUND: Extracranial internal carotid artery (eICA) tortuosity may trigger cerebral ischemia, and body mass index (BMI) is a measure of body mass based on height and weight. The main purpose of this study is to determine the influence of BMI on the tortuosity of eICA. METHODS: A total of 926 carotid artery angiograms were performed in 513 patients, of which 116 cases and matched controls were selected. Arterial tortuosity was defined as simple tortuosity, kinking, or coiling. The severity of tortuosity was measured by tortuosity index, formula: [(actual length/straight-line length - 1) × 100]. RESULTS: BMIs were different between the two groups [tortuosity: 27.06 kg/m2 (SD 2.81 kg/m2) versus none: 23.3 kg/m2 (SD 2.78 kg/m2); p < 0.001]. BMI was independently and significantly associated with eICA tortuosity (odds ratio 1.59; 95% confidence interval, 1.35-1.86; p < 0.001). eICA tortuosity index is linearly associated with BMI (exponential coefficient ß = 1.067, p < 0.001). The optimal predictive threshold of BMI for eICA tortuosity was 25.04 kg/m2. The physiological mechanism underlying the reasons why higher BMI has negative influence on extracranial carotid artery tortuosity may be an intra-abdominal hypertension caused by a much higher amount of body fat stored in visceral adipose tissue. CONCLUSION: Our result reveals a novel role for greater BMI on the presence of eICA tortuosity. For each increase in BMI of 1 kg/m2, there is a corresponding 1.59-fold increase in the risk of developing eICA tortuosity. The severity of eICA tortuosity increases linearly with increased BMI.

Total glucosides of paeony inhibits lipopolysaccharide-induced proliferation, migration and invasion in androgen insensitive prostate cancer cells.

Previous studies demonstrated that inflammatory microenvironment promoted prostate cancer progression. This study investigated whether total glucosides of paeony (TGP), the active constituents extracted from the root of Paeonia Lactiflora Pall, suppressed lipopolysaccharide (LPS)-stimulated proliferation, migration and invasion in androgen insensitive prostate cancer cells. PC-3 cells were incubated with LPS (2.0 µg/mL) in the absence or presence of TGP (312.5 µg /mL). As expected, cells at S phase and nuclear CyclinD1, the markers of cell proliferation, were increased in LPS-stimulated PC-3 cells. Migration activity, as determined by wound-healing assay and transwell migration assay, and invasion activity, as determined by transwell invasion assay, were elevated in LPS-stimulated PC-3 cells. Interestingly, TGP suppressed LPS-stimulated PC-3 cells proliferation. Moreover, TGP inhibited LPS-stimulated migration and invasion of PC-3 cells. Additional experiment showed that TGP inhibited activation of nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK)/p38 in LPS-stimulated PC-3 cells. Correspondingly, TGP attenuated upregulation of interleukin (IL)-6 and IL-8 in LPS-stimulated PC-3 cells. In addition, TGP inhibited nuclear translocation of signal transducer and activator of transcription 3 (STAT3) in LPS-stimulated PC-3 cells. These results suggest that TGP inhibits inflammation-associated STAT3 activation and proliferation, migration and invasion in androgen insensitive prostate cancer cells.

Low vitamin D status is associated with inflammation in patients with prostate cancer.

Vitamin D deficiency has been associated with increased risks of prostate cancer. Nevertheless, the mechanisms remain unclear. The aim of this study was to analyze the association among prostate cancer, vitamin D status and inflammation. Sixty patients with newly diagnosed prostate cancer and 120 age-matched controls were recruited for this study. Vitamin D status was evaluated and serum inflammatory molecules were measured. Serum 25-(OH)D was lower in patients with prostate cancer. Moreover, serum 25(OH)D was lower in patients with severe prostate cancer than patients with mild and moderate prostate cancer. By contrast, serum C-reactive protein (CRP) and interleukin (IL)-8, two inflammatory molecules, were elevated in patients with prostate cancer. Serum 25-(OH)D was negatively correlated with serum CRP and IL-8 in patients with prostate cancer. Additional analysis showed that the percentage of vitamin D receptor positive nucleus in the prostate was reduced in patients with prostate cancer. By contrast, the percentage of nuclear factor kappa B p65-positive nucleus was elevated in patients with prostate cancer. Our results provide evidence that there is an association among prostate cancer, vitamin D deficiency and inflammatory signaling. Inflammation may be an important mediator for prostate cancer progression in patients with low vitamin D status.