Rabeprazole mitigates obesity-induced chronic inflammation and insulin resistance associated with increased M2-type macrophage polarization.

Macrophage polarization is closely associated with obesity-induced chronic inflammation and insulin resistance. Proton pump inhibitor Rabeprazole has long been used to treat gastritis and gastric ulcers. However, whether Rabeprazole plays a role in macrophage polarization during obesity is unknown. Here, we show that Rabeprazole suppresses M1-type macrophage-mediated inflammation, leads to increased M2-type macrophages and alters the polarization status from M1 to M2 in vitro. Mechanistically, Rabe-regulated macrophage polarization is associated with inhibition of NF-κB and activation of STAT6 signaling pathways. Furthermore, Rabeprazole induces M2-type adipose tissue macrophages and alleviates chronic inflammation, improving glucose tolerance and insulin sensitivity in high-fat diet-fed mice. In addition, Rabeprazole increases CD206+ M2-type liver macrophages and relieves liver inflammation, alleviating liver injury and lipid accumulation. Thus, our findings show that Rabeprazole effectively regulates macrophage polarization and controls obesity-associated chronic inflammation and insulin resistance, thus providing a potential therapeutic strategy against obesity-associated metabolic diseases.

Diagnostic and prognostic value of plasma miR-106a-5p levels in patients with acute heart failure.

BACKGROUND: It is essential to find reliable biomarkers for early diagnosis and prognosis of acute heart failure (AHF) for its mitigation. Currently, increasing attention is paid to the role of microRNAs (miRNAs/miRs) as diagnostic or prognostic markers for cardiovascular diseases. Since plasma miR-106a-5p has been observed to be downregulated in AHF, its value in the diagnosis and prognostic assessment of AHF deserves further exploration. Accordingly, this study analyzed the diagnostic and prognostic value of plasma miR-106a-5p in AHF patients. METHODS: Prospectively, this study included 127 AHF patients who met the 2021 European Society of Cardiology Guidelines and 127 control individuals. Plasma miR-106a-5p levels were determined with RT-qPCR. Spearman correlation analysis was performed to evaluate the correlation of plasma miR-106a-5p levels with NT-proBNP and hs-CRP levels in AHF patients. All AHF patients were followed up for 1 year and allocated into poor and good prognosis groups, and plasma miR-106a-5p levels were compared. The diagnostic and prognostic value of plasma miR-106a-5p for AHF was assessed with a receiver-operating characteristic curve. RESULTS: Plasma miR-106a-5p was lowly expressed in AHF patients versus controls (0.53 ± 0.26 vs. 1.09 ± 0.46) and showed significant negative correlations with NT-proBNP and hs-CRP levels. Plasma miR-106a-5p level < 0.655 could assist in AHF diagnosis. Plasma miR-106a-5p levels were markedly lower in poor-prognosis AHF patients than in good-prognosis patients. Plasma miR-106a-5p level < 0.544 could assist in predicting poor prognosis in AHF patients. CONCLUSION: Plasma miR-106a-5p is downregulated in AHF patients and could assist in diagnosis and poor prognosis prediction of AHF.

CD177 is a novel IgG Fc receptor and CD177 genetic variants affect IgG-mediated function.

CD177 plays an important role in the proliferation and differentiation of myeloid lineage cells including neutrophils, myelocytes, promyelocytes, megakaryocytes, and early erythroblasts in bone marrow. CD177 deficiency is a common phenotype in humans. Our previous studies revealed genetic mechanisms of human CD177 deficiency and expression variations. Up to now, immune functions of CD177 remain undefined. In the current study, we revealed human IgG as a ligand for CD177 by using flow cytometry, bead-rosette formation, and surface plasmon resonance (SPR) assays. In addition, we show that CD177 variants affect the binding capacity of CD177 for human IgG. Furthermore, we showed that the CD177 genetic variants significantly affect antibody-dependent cell-mediated cytotoxicity (ADCC) function. The demonstration of CD177 as a functional IgG Fc-receptor may provide new insights into CD177 immune function and genetic mechanism underlying CD177 as biomarkers for human diseases.

Statin-mediated reduction in mitochondrial cholesterol primes an anti-inflammatory response in macrophages by upregulating JMJD3.

Stains are known to be anti-inflammatory, but the mechanism remains poorly understood. Here we show that macrophages, either treated with statin in vitro or from statin-treated mice, have reduced cholesterol levels and higher expression of Jmjd3, a H3K27me3 demethylase. We provide evidence that lowering cholesterol levels in macrophages suppresses the ATP synthase in the inner mitochondrial membrane (IMM) and changes the proton gradient in the mitochondria. This activates NFkB and Jmjd3 expression to remove the repressive marker H3K27me3. Accordingly, the epigenome is altered by the cholesterol reduction. When subsequently challenged by the inflammatory stimulus LPS (M1), both macrophages treated with statins in vitro or isolated from statin-treated mice in vivo, express lower levels pro-inflammatory cytokines than controls, while augmenting anti-inflammatory Il10 expression. On the other hand, when macrophages are alternatively activated by IL4 (M2), statins promote the expression of Arg1, Ym1, and Mrc1. The enhanced expression is correlated with the statin-induced removal of H3K27me3 from these genes prior to activation. In addition, Jmjd3 and its demethylase activity are necessary for cholesterol to modulate both M1 and M2 activation. We conclude that upregulation of Jmjd3 is a key event for the anti-inflammatory function of statins on macrophages.

Safety assessment, whole genome sequence, and metabolome analysis of Streptococcus thermophilus CICC 20372 for bone cement fermentation.

Bone is a kind of meat processing by-product with high nutritional value but low in calorie, which is a typical food in China and parts of East Asian countries. Microbial fermentation by lactic acid bacteria showed remarkable advantages to increase the absorption of nutrients from bone cement by human body. Streptococcus thermophilus CICC 20372 is proven to be a good starter for bone cement fermentation. No genes encoding virulence traits or virulence factors were found in the genome of S. thermophilus CICC 20372 by a thorough genomic analysis. Its notable absence of antibiotic resistance further solidifies the safety. Furthermore, the genomic analysis identified four types of gene clusters responsible for the synthesis of antimicrobial metabolites. A comparative metabolomic analysis was performed by cultivating the strain in bone cement at 37 °C for 72 h, with the culture in de Man, Rogosa, and Sharpe (MRS) medium as control. Metabolome analysis results highlighted the upregulation of pathways involved in 2-oxocarboxylic acid metabolism, ATP-binding cassette (ABC) transporters, amino acid synthesis, and nucleotide metabolism during bone cement fermentation. S. thermophilus CICC 20372 produces several metabolites with health-promoting function during bone cement fermentation, including indole-3-lactic acid, which is demonstrated ameliorative effects on intestinal inflammation, tumor growth, and gut dysbiosis. In addition, lots of nucleotide and organic acids were accumulated at higher levels, which enriched the fermented bone cement with a variety of nutrients. Collectively, these features endow S. thermophilus CICC 20372 a great potential strain for bone food processing.

Helical insertion of polyphenylene chains into confined cylindrical slits composed of two carbon nanotubes.

The helical insertion behavior of poly(para-phenylene) (PP) chains into confined cylindrical slits constructed by two carbon nanotubes (CNTs) with different diameters is studied by molecular dynamics simulations. The contribution of system energy and each energy component to helical self-assembly is discussed to further explain the conditions, driving force and mechanism. The width and length of the slit, the diameter of the outer tube and the temperature have a great impact on the helical insertion of PP chains. Two equations are proposed to confirm the diameter and the distances between the PP helix and the inner and outer walls of the given CNTs. The helical self-assembly of PP with different numbers of chains inserted into the slits is further studied. This study has a great benefit in understanding the conformational behavior of polymers, even biological macromolecules in confinements.

14-3-3 proteins regulate the HCHO stress response by interacting with AtMDH1 and AtGS1 in tobacco and Arabidopsis.

Formaldehyde (HCHO) is one of the most essential common carcinogenic environmental pollutants. While 14-3-3 proteins are known to regulate the response of plants to HCHO stress, the regulatory mechanisms responsible for a tolerant phenotype remain unclear. We first performed qPCR analysis of HCHO-treated Arabidopsis and tobacco and determined that the expression of At14-3-3PSI and Nt14-3-3C genes was rapidly upregulated after HCHO stress. Furthermore, overexpression of 14-3-3, AtMDH1 or AtGS1 genes enhanced plant HCHO absorption capacity and resistance, and knockdown or knockout of 14-3-3, AtMDH1 or AtGS1 genes reduced plant HCHO absorption capacity and resistance. However, overexpression of the AtGS1 and AtMDH1 genes in the At14-3-3 psi mutant restored HCHO uptake and resistance in Arabidopsis. Moreover, 14-3-3 bound to the N-terminus of AtMDH1 and the C-terminus of AtGS1, respectively, and repressed and enhanced their expression. The 13C NMR results of HCHO stress mutants Atgs1 and Atmdh1 showed that the metabolites Glu and Asp rapidly increased, indicating that AtGS1 and AtMDH1 were indeed indispensable for Arabidopsis to metabolize HCHO. In conclusion, we uncovered a HCHO stress response mechanism mediated by 14-3-3, which enhances the plant's ability to absorb HCHO, deepening our understanding of how plants respond to HCHO stress.

Identification and validation of a major quantitative trait locus for spike length and compactness in the wheat (Triticum aestivum L.) line Chuanyu12D7.

Spike length (SL) and spike compactness (SC) are crucial traits related to wheat (Triticum aestivum L.) yield potential. In this study, a backcrossed inbred lines (BILs) population segregating for SL/SC was developed by using a commercial variety chuanyu25 as recurrent parent and a backbone parent Chuanyu12D7. Bulked segregant analysis (BSA) combined with the Wheat 660K SNP array was performed to conduct quantitative trait locus (QTL) mapping. A major and stable SL/SC QTL (designated as QSl/Sc.cib-2D.1) was identified on chromosome 2DS, explaining 45.63-59.72% of the phenotypic variation. QSl/Sc.cib-2D.1 was mapped to a 102.29-Kb interval by flanking SNPs AX-110276364 and AX-111593853 using a BC4F2:3 population. Since QSl/Sc.cib-2D.1 is linked to the Rht8 gene, their additive effects on plant type and spike type were analysed. Remarkably, the superior allele of QSl/Sc.cib-2D.1 combined with Rht8 can increase SL and TGW, and decrese SC without any apparent trade-offs in other yield-related traits. In addition, the closely linked kompetitive allele-specific PCR (KASP) markers of this locus were developed for marker-assisted selection (MAS) breeding. Four genes within the physical interval were considered as potential candidates based on expression patterns as well as orthologous gene functions. These results laid the foundation for map-based cloning of the gene(s) underlying QSl/Sc.cib-2D.1 and its potential application in wheat ideotype breeding.

Study on the active ingredients and mechanism of action of Jiaotai Pill in the treatment of type 2 diabetes based on network pharmacology: A review.

To explore the potential active ingredients and related mechanisms of Jiaotai Pill in the treatment of Type 2 diabetes mellitus (T2DM) based on network pharmacology and molecular docking. The main active components of Jiaotai Pills were obtained by TCMSP and BATMAN-TCM database combined with literature mining, and the targets of the active components of Jiaotai Pills were predicted by reverse pharmacophore matching (PharmMapper) method. Verifying and normalizing the obtained action targets by using a Uniprot database. Obtaining T2DM related targets through GeneCards, the online mendelian inheritance in man, DrugBank, PharmGKB and therapeutic target databases, constructing a Venn diagram by using a Venny 2.1 online drawing platform to obtain the intersection action targets of Jiaotai pills and T2DM, and the protein-protein interaction network was constructed by String platform. Bioconductor platform and R language were used to analyze the function of gene ontology and the pathway enrichment of Kyoto Encyclopedia of Genes and Genomes. A total of 21 active components and 262 potential targets of Jiaotai Pill were screened by database analysis and literature mining, including 89 targets related to T2DM. Through gene ontology functional enrichment analysis, 1690 biological process entries, 106 molecular function entries and 78 cellular component entries were obtained. Seven pathways related to T2DM were identified by Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. Jiaotai Pill can achieve the purpose of treating T2DM through multiple active ingredients, multiple disease targets, multiple biological pathways and multiple pathways, which provides a theoretical basis for the clinical treatment of T2DM by Jiaotai Pill.

Identification and Quantification of Dimethachlon Degradation Products in Soils and Their Effects on Soil Enzyme Activities.

In this study, high-performance liquid chromatography-high-resolution mass spectrometry (HPLC-HRMS, Q-Exactive Orbitrap) and Compound Discoverer 3.3 were used to screen dimethachlon degradation products in soils. Four metabolites 4-(3,5-dichloroanilino)-4-oxobutanoic acid (DCBAA), 3,5-dichloroaniline (3,5-DCA), succinic acid, and muconic acid were confirmed by primary and secondary ion mass spectrometry comparisons between standards and samples. A quantitative analysis method of dimethachlon residues and four metabolites in soils was developed using HPLC-HRMS. Dimethachlon degradation in agricultural soil indoor unsterilized, sterilized, and field environments in three typical areas was measured. Dimethachlon degraded fast with a half-life of less than 1 day in three nonsterile soils. The maximum DCBAA and 3,5-DCA residues during degradation could reach 22.5-35.2% of the initial concentration of the parent dimethachlon. The metabolite DCBAA had a greater impact on soil enzyme activity than the parent dimethachlon.

In vivo characterization of magnetic resonance imaging-based T1w/T2w ratios reveals myelin-related changes in temporal lobe epilepsy.

Temporal lobe epilepsy (TLE) is the most common type of intractable epilepsy in adults. Although brain myelination alterations have been observed in TLE, it remains unclear how the myelination network changes in TLE. This study developed a novel method in characterization of myelination structural covariance network (mSCN) by T1-weighted and T2-weighted magnetic resonance imaging (MRI). The mSCNs were estimated in 42 left TLE (LTLE), 42 right TLE (RTLE) patients, and 41 healthy controls (HCs). The topology of mSCN was analyzed by graph theory. Voxel-wise comparisons of myelination laterality were also examined among the three groups. Compared to HC, both patient groups showed decreased myelination in frontotemporal regions, amygdala, and thalamus; however, the LTLE showed lower myelination in left medial temporal regions than RTLE. Moreover, the LTLE exhibited decreased global efficiency compared with HC and more increased connections than RTLE. The laterality in putamen was differently altered between the two patient groups: higher laterality at posterior putamen in LTLE and higher laterality at anterior putamen in RTLE. The putamen may play a transfer station role in damage spreading induced by epileptic seizures from the hippocampus. This study provided a novel workflow by combination of T1-weighted and T2-weighted MRI to investigate in vivo the myelin-related microstructural feature in epileptic patients first time. Disconnections of mSCN implicate that TLE is a system disorder with widespread disruptions at regional and network levels.

An accurate genetic assay to identify human neutrophil antigen 2 deficiency.

OBJECTIVE: We aimed to develop accurate and user-friendly genetic assays to identify the inherited neutrophil antigen-2 (HNA-2) deficiency in humans. BACKGROUND: HNA-2 is one of the most important neutrophil antigens implicated in a number of human disorders. HNA-2 deficiency or HNA-2 null is a common phenotype observed in 3%-5% Americans. HNA-2 null individuals are at risk to produce isoantibodies (or alloantibodies) that play important roles in transfusion-related acute lung injury, immune neutropenia, and bone marrow graft failure. We previously demonstrated that the CD177 coding SNP 787A > T (c.787A > T) is the most important genetic determinant for HNA-2 deficiency. However, reliable genetic assays are not available for routine clinical laboratory application up to now. STUDY DESIGN AND METHODS: A novel polymerase chain reaction (PCR) strategy was used to determine genotypes of the CD177 SNP c.787A > T. In the simplified PCR assay, all allele specific primers and internal control primers were included in the same reaction, which ensures reliability of the assay. In addition, a novel high-throughput nested TaqMan assay was developed to determine genotypes of c.787A > T for large population genetic analysis of HNA-2 deficiency. RESULTS: CD177 SNP c787A > T genotypes of 396 subjects were 100% concordant among the single PCR reaction method, the nested TaqMan assay, and Sanger Sequencing analysis. Out of 396 subjects, all 18 donors with the CD177 STP homozygous genotype were HNA-2 null. CONCLUSION: The novel PCR-based genotyping assay is accurate to identify HNA-2 deficient individuals and is suitable for clinical laboratories. In addition, the innovative high-throughput nested TaqMan assay will be useful for large-scale population screens and genetic studies of HNA-2 deficiency.

Accumulation, metabolites formation and elimination behavior of rac-glufosinate-ammonium and glufosinate-P in zebrafish (Danio rerio).

An efficient trace detection method for the determination of residues of the glufosinate enantiomers and metabolites in zebrafish by HPLC-Q-Exactive Orbitrap Mass Spectrometry was developed. After the purification of dichloromethane and Oasis PRiME HLB SPE column, the recovery ranges from 77% to 104%, with RSD < 10.03%. The limits of quantitation in zebrafish were 0.006-0.02 mg/kg. The results revealed zebrafish absorbed glufosinate slowly, reaching a steady state in 10-14 days, and the bioaccumulation factor (BCF) of D/L-glufosinate-ammonium was less than 0.3. L-glufosinate-ammonium accumulated preferentially in zebrafish. The residue of the metabolite N-acetyl glutamate (NAG) was smaller than that of 3-methyl phosphonic acid (MPP). D/L-glufosinate-ammonium had an elimination half-life of less than 2.3 days during the elimination phase. The bioaccumulation and elimination behavior of glufosinate-ammonium in zebrafish aquatic system was shown in this work, which offered scientific data for assessing the food safety of rac-glufosinate-ammonium and glufosinate-P (pure L-glufosinate-ammonium) in fish.

Kinetics of the photolysis of pyridaben and its main photoproduct in aqueous environments under simulated solar irradiation.

The photolytic fate of pyridaben and its main photolysis product was investigated in different aqueous solutions. Results showed that the photolysis of pyridaben followed pseudo first-order kinetics or the hockey-stick model. In buffer solutions, the half-life of pyridaben was the shortest at pH 4, while the degradation rate within 24 h was the highest at pH 9. Humic acids (HA) at concentrations of 1-20 mg L-1 favored the photolysis of pyridaben while fulvic acids (FA) did not have a significant effect. Nitrate at low concentrations (0.01 mM) accelerated the photolysis and Fe(iii) at high concentrations (0.01 and 0.1 mM) significantly inhibited the photolysis. The photolysis rate of pyridaben in rainwater, tap water, and river water was significantly higher than that in distilled water. The half-lives in distilled water, rainwater, tap water, river water, and pond water were 2.36, 1.36, 1.61, 1.77, and 2.68 h, respectively. Ultra-high-performance liquid chromatography/high-resolution mass spectrometry identified M328 as a photolysis product. The degradation of M328 followed pseudo first-order kinetics in distilled water, buffer solutions and aqueous solutions fortified with HA. The half-lives of M328 were in the range of 7.07-13.95 h. These results are essential for further environmental risk assessment of pyridaben.

Effect of Vitamin D Combined with Recombinant Human Growth Hormone in Children with Growth Hormone Deficiency.

Objective: Growth hormone deficiency (GHD) refers to the complete or partial lack of pituitary growth hormone synthesis and secretion. This study is aimed at investigating the efficacy of vitamin D and recombinant human growth hormone (rhGH) in children with GHD. Methods: A total of 100 children with GHD at our hospital were included between 1st January 2018 and 31st October 2020. The patients were divided into a study group (n = 70, received vitamin D combined with rhGH) and a control group (n = 30, received rhGH). The growth and development (bone age, growth rate, and height), bone metabolism (bone alkaline phosphatase (BAP), ß-collagen degradation product (ß-CTX), osteocalcin (OC), and amino-terminal propeptide type I procollagen (PINP)), insulin-like growth factor 1 (IGF-1), ghrelin, and adverse reactions in the two groups were measured before and 12 months after treatment. Results: There were no significant differences in the bone age, growth rate, and height between the two groups before treatment. After 12 months of treatment, the bone age, growth rate, and height of the study group were significantly higher than those of the control group. After 12 months of treatment, the levels of serum BAP, PINP, and OC in the study group were significantly higher than those in the control group, while the levels of ß-CTX in the study group were significantly lower than those in the control group. The serum IGF-1 level in the study group was significantly higher than that in the control group, while the ghrelin level in the study group was lower. There was no significant difference in the incidence of adverse reactions between the two groups. Conclusion: Combined rhGH and vitamin D treatment can promote growth and development, improve bone metabolism, and regulate IGF-1 and ghrelin levels.

Novel Human FCGR1A Variants Affect CD64 Functions and Are Risk Factors for Sarcoidosis.

CD64 (or FcγRIA) is the sole functional high affinity IgG Fc receptor coded by FCGR1A gene in humans. The FCGR1A genetics has not been comprehensively investigated and effects of human FCGR1A variants on immune functions remain unknown. In the current study, we identified three novel FCGR1A variants including the single nucleotide variant (SNV) rs1848781 (c.-131) in the proximal FCGR1A gene promoter region, the rs587598788 indel variant within the FCGR1A intron 5, and the non-synonymous SNV rs1050204 (c.970G>A or FcγRIA-p.D324N) in the FCGR1A coding region. Genotype-phenotype analyses revealed that SNV rs1848781 genotypes were significantly associated with CD64 expression levels. Promoter reporter assays show that rs1848781G allele had significantly higher promoter activity than the rs1848781C, confirming that the rs1848781 is a functional FCGR1A SNV affecting promoter activity and gene expression. The rs587598788 indel genotypes were also significantly associated with levels of CD64 expression. Moreover, the non-synonymous SNV rs1050204 (FcγRIA-p.D324N) alleles significantly affected CD64-mediated phagocytosis, degranulation, and pro-inflammatory cytokine productions. Genetic analyses revealed that FCGR1A genotypes were significantly associated with sarcoidosis susceptibility and severity. Our data suggest that FCGR1A genetic variants may affect immune responses and play a role in sarcoidosis.

Examination of IgG Fc Receptor CD16A and CD64 Expression by Canine Leukocytes and Their ADCC Activity in Engineered NK Cells.

Human natural killer (NK) cells can target tumor cells in an antigen-specific manner by the recognition of cell bound antibodies. This process induces antibody-dependent cell-mediated cytotoxicity (ADCC) and is exclusively mediated by the low affinity IgG Fc receptor CD16A (FcγRIIIA). Exploiting ADCC by NK cells is a major area of emphasis for advancing cancer immunotherapies. CD64 (FcγRI) is the only high affinity IgG FcR and it binds to the same IgG isotypes as CD16A, but it is not expressed by human NK cells. We have generated engineered human NK cells expressing recombinant CD64 with the goal of increasing their ADCC potency. Preclinical testing of this approach is essential for establishing efficacy and safety of the engineered NK cells. The dog provides particular advantages as a model, which includes spontaneous development of cancer in the setting of an intact and outbred immune system. To advance this immunotherapy model, we cloned canine CD16A and CD64 and generated specific mAbs. We report here for the first time the expression patterns of these FcγRs on dog peripheral blood leukocytes. CD64 was expressed by neutrophils and monocytes, but not lymphocytes, while canine CD16A was expressed at high levels by a subset of monocytes and lymphocytes. These expression patterns are similar to that of human leukocytes. Based on phenotypic characteristics, the CD16A+ lymphocytes consisted of T cells (CD3+ CD8+ CD5dim α/ß TCR+) and NK cells (CD3- CD5- CD94+), but not B cells. Interestingly, the majority of canine CD16A+ lymphocytes were from the T cell population. Like human CD16A, canine CD16A was downregulated by a disintegrin and metalloproteinase 17 (ADAM17) upon leukocyte activation, revealing a conserved means of regulation. We also directly demonstrate that both canine CD16A and CD64 can induce ADCC when expressed in the NK cell line NK-92. These findings pave the way to engineering canine NK cells or T cells with high affinity recombinant canine CD64 to maximize ADCC and to test their safety and efficacy to benefit both humans and dogs.

Comparing toxicity and biodegradation of racemic glufosinate and L-glufosinate in green algae Scenedesmus obliquus.

Glufosinate-ammonium, a widely used chiral herbicide, has become the focus of attention because of its toxicity toward non-target organisms and its degradation behavior in the environment. With the introduction of L-glufosinate-ammonium products, the toxicity and environmental behavior of rac-glufosinate-ammonium and L-glufosinate-ammonium have become the subject of increasing interest. The overall goal of this study was to investigate the differences in toxicity and biodegradation of rac-glufosinate-ammonium and L-glufosinate-ammonium in an aquatic organism, Scenedesmus obliquus. The toxicity of rac-glufosinate-ammonium and L-glufosinate-ammonium to S. obliquus was compared by measuring EC50, malondialdehyde (MDA) content, protein content and antioxidant enzyme activity. The 96-h EC50 values of rac-glufosinate-ammonium and L-glufosinate-ammonium were 57.22 µg/mL and 25.55 µg/mL, respectively, which indicated that L-glufosinate-ammonium was more toxic to S. obliquus than rac-glufosinate-ammonium. Based on the MDA content, protein content, and antioxidant enzyme (SOD and CAT) activity, we found that L-glufosinate-ammonium could cause more serious oxidative damage than rac-glufosinate-ammonium. The residual amount of glufosinate-ammonium and its metabolites in the culture medium and S. obliquus were determined by HPLC-HRMS. Comparison of glufosinate-ammonium concentrations in algae-free and algae-containing media, showed that glufosinate-ammonium degradation in the S. obliquus system was significantly increased, and the degradation rate of L-glufosinate-ammonium was faster than that of D-glufosinate-ammonium. No enantiomerization was observed for pure L-glufosinate-ammonium treatment. N-acetyl-glufosinate was identified as the main metabolite of glufosinate-ammonium.

Identification of differentially expressed genes using microarray analysis and COL6A1 induction of bone metastasis in non-small cell lung cancer.

Non-small cell lung cancer (NSCLC) is a major cause of cancer-associated mortality worldwide, and bone metastasis is the most prevalent event observed in patients with advanced NSCLC. However, the pathogenesis of bone metastases has not been fully elucidated. In the present study, differentially expressed genes (DEGs) were identified by gene expression microarray analysis of NSCLC tissue samples with or without bone metastases. Subsequently, collagen type 6A1 (COL6A1) was chosen as the target gene through Ingenuity Pathway Analysis and reverse transcription-quantitative (RT-q) PCR validation of the top eight DEGs. COL6A1 was overexpressed or knocked down, and the proliferation and invasion of NSCLC cells was assessed using Cell Counting Kit-8, colony formation and Transwell invasion assays. Additionally, the osteogenic capacity of HOB and hES-MP 002.5 cells was assessed using RT-qPCR, western blotting, Alizarin Red and alkaline phosphatase staining. A total of 364 DEGs were identified in NSCLC tissues with bone metastases compared with NSCLC tissues without bone metastases, including 140 upregulated and 224 downregulated genes. Gene Ontology analysis results demonstrated that the upregulated and downregulated genes were primarily enriched in 'cellular process', 'metabolic process' and 'biological regulation'. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis revealed that the upregulated genes were primarily enriched in 'cysteine and methionine metabolism', 'oxidative phosphorylation' and 'ribosome', whereas the downregulated genes were primarily enriched in the 'transcriptional misregulation in cancer', 'ribosome' and 'mitophagy-animal' pathways. COL6A1 was highly expressed in NSCLC tissue samples with bone metastases. Functionally, COL6A1 overexpression induced the proliferation and invasion of HARA cells, and its knockdown inhibited the proliferation and invasion of HARA-B4 cells. Finally, it was demonstrated that HOB and hES-MP 002.5 cells exhibited osteogenic capacity, and overexpression of COL6A1 in HARA cells increased the adhesion of these cells to the osteoblasts, whereas knockdown of COL6A1 in HARA-B4 cells reduced their adhesive ability. In conclusion, COL6A1 may serve as a potential diagnostic marker and therapeutic target for bone metastasis in NSCLC.