Phenotypic comparison and the potential antitumor function of immortalized bone marrow-derived macrophages (iBMDMs).

Introduction: Macrophages are an important component of innate immunity and involved in the immune regulation of multiple diseases. The functional diversity and plasticity make macrophages to exhibit different polarization phenotypes after different stimuli. During tumor progression, the M2-like polarized tumor-associated macrophages (TAMs) promote tumor progression by assisting immune escape, facilitating tumor cell metastasis, and switching tumor angiogenesis. Our previous studies demonstrated that functional remodeling of TAMs through engineered-modifying or gene-editing provides the potential immunotherapy for tumor. However, lack of proliferation capacity and maintained immune memory of infused macrophages restricts the application of macrophage-based therapeutic strategies in the repressive tumor immune microenvironment (TIME). Although J2 retrovirus infection enabled immortalization of bone marrow-derived macrophages (iBMDMs) and facilitated the mechanisms exploration and application, little is known about the phenotypic and functional differences among multi kinds of macrophages. Methods: HE staining was used to detect the biosafety of iBMDMs, and real-time quantitative PCR, immunofluorescence staining, and ELISA were used to detect the polarization response and expression of chemokines in iBMDMs. Flow cytometry, scratch assay, real-time quantitative PCR, and crystal violet staining were used to analyze its phagocytic function, as well as its impact on tumor cell migration, proliferation, and apoptosis. Not only that, the inhibitory effect of iBMDMs on tumor growth was detected through subcutaneous tumor loading, while the tumor tissue was paraffin sectioned and flow cytometry was used to detect its impact on the tumor microenvironment. Results: In this study, we demonstrated iBMDMs exhibited the features of rapid proliferation and long-term survival. We also compared iBMDMs with RAW264.7 cell line and mouse primary BMDMs with in vitro and in vivo experiments, indicating that the iBMDMs could undergo the same polarization response as normal macrophages with no obvious cellular morphology changes after polarization. What's more, iBMDMs owned stronger phagocytosis and pro-apoptosis functions on tumor cells. In addition, M1-polarized iBMDMs could maintain the anti-tumor phenotypes and domesticated the recruited macrophages of receptor mice, which further improved the TIME and repressed tumor growth. Discussion: iBMDMs can serve as a good object for the function and mechanism study of macrophages and the optional source of macrophage immunotherapy.

Identification and validation of candidate clinical signatures of apolipoprotein L isoforms in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a lethal malignancy worldwide with an increasing number of new cases each year. Apolipoprotein (APOL) isoforms have been explored for their associations with HCC.The GSE14520 cohort was used for training data; The Cancer Genome Atlas (TCGA) database was used for validated data. Diagnostic, prognostic significance and mechanisms were explored using these cohorts. Risk score models and nomograms were constructed using prognosis-related isoforms and clinical factors for survival prediction. Oncomine and HCCDB databases were further used for validation of diagnostic, prognostic significance. APOL1, 3, and 6 were differentially expressed in two cohorts (all P ≤ 0.05). APOL1 and APOL6 had diagnostic capacity whereas APOL3 and APOL6 had prognostic capacity in two cohorts (areas under curves [AUCs] > 0.7, P ≤ 0.05). Mechanism studies demonstrated that APOL3 and APOL6 might be involved in humoral chemokine signaling pathways (all P ≤ 0.05). Risk score models and nomograms were constructed and validated for survival prediction of HCC. Moreover, diagnostic values of APOL1 and weak APOL6 were validated in Oncomine database (AUC > 0.700, 0.694); prognostic values of APOL3 and APOL6 were validated in HCCDB database (all P < 0.05). Differentially expressed APOL1 and APOL6 might be diagnostic biomarkers; APOL3 and APOL6 might be prognostic biomarkers of RFS and OS for HCC via chemokine signaling pathways.

Clinical implications of RAB13 expression in pan-cancer based on multi-databases integrative analysis.

Worldwide, cancer is a huge burden, and each year sees an increase in its incidence. RAB (Ras-related in brain) 13 is crucial for a number of tumor types. But more research on RAB13's tumor-related mechanism is still required. This study's goal was to investigate RAB13's function in human pan-cancer, and we have also preliminarily explored the relevant mechanisms. To investigate the differential expression, survival prognosis, immunological checkpoints, and pathological stage of RAB13 in human pan-cancer, respectively, databases of TIMER2.0, GEPIA 2, and UALCAN were employed. CBioPortal database was used to analyze the mutation level, meanwhile, PPI network was constructed based on STRING website. The putative functions of RAB13 in immunological infiltration were investigated using single sample gene set enrichment analysis (ssGSEA). The mechanism of RAB13 in hepatocellular cancer was also briefly investigated by us using gene set enrichment analysis (GSEA). RAB13 was differentially expressed in a number of different cancers, including liver hepatocellular carcinoma (LIHC), stomach adenocarcinoma (STAD), etc. Additionally, RAB13 overexpression in LGG and LIHC is associated with a worse prognosis, including overall survival (OS) and disease-free survival (DFS). Then, we observed that early in BLCA, BRAC, CHOL, ESCA, HNSC, KICH, KIRC, LIHC, LUAD, LUSC, and STAD, the level of RAB13 expression was raised. Next, we found that "amplification" was the most common mutation in RAB13. The expression of SLC39A1, JTB, SSR2, SNAPIN, and RHOC was strongly positively linked with RAB13, according to a correlation study. RAB13 favorably regulated B cell, CD8 + T cell, CD4 + T cell, macrophage, neutrophil, and dendritic cell in LIHC, according to immune infiltration analysis. Immune checkpoint study revealed a positive correlation between RAB13 expression and PD1, PDL1, and CTLA4 in LIHC. According to GSEA, RAB13 is involved in a number of processes in LIHC, including MTORC1 signaling, MYC targets v1, G2M checkpoint, MITOTIC spindle, DNA repair, P53 pathway, glycolysis, PI3K-AKT-MTOR signaling, etc. RAB13 is a possible therapeutic target in LIHC and can be used as a prognostic marker.

Mechanisms and regulations of ferroptosis.

Regulation of cell mortality for disease treatment has been the focus of research. Ferroptosis is an iron-dependent regulated cell death whose mechanism has been extensively studied since its discovery. A large number of studies have shown that regulation of ferroptosis brings new strategies for the treatment of various benign and malignant diseases. Iron excess and lipid peroxidation are its primary metabolic features. Therefore, genes involved in iron metabolism and lipid metabolism can regulate iron overload and lipid peroxidation through direct or indirect pathways, thereby regulating ferroptosis. In addition, glutathione (GSH) is the body's primary non-enzymatic antioxidants and plays a pivotal role in the struggle against lipid peroxidation. GSH functions as an auxiliary substance for glutathione peroxidase 4 (GPX4) to convert toxic lipid peroxides to their corresponding alcohols. Here, we reviewed the researches on the mechanism of ferroptosis in recent years, and comprehensively analyzed the mechanism and regulatory process of ferroptosis from iron metabolism and lipid metabolism, and then described in detail the metabolism of GPX4 and the main non-enzymatic antioxidant GSH in vivo.

Clinical efficacy of botulinum toxin type A in the treatment of fasciitis pain: A systematic review and meta-analysis.

BACKGROUND: The purpose of this meta-analysis was to assess the effectiveness of botulinum toxin type A (BoNT-A) in reducing pain associated with fasciitis. By synthesizing the findings from multiple studies, we aimed to provide a comprehensive evaluation of the current evidence regarding the efficacy of BoNT-A in the treatment of fasciitis pain. METHODS: To identify studies for our report, we conducted electronic database searches of Embase, PubMed, Web of Science, and the Cochrane Library from their inception to November 20, 2022. We included only randomized controlled trials that examined the therapeutic effects of BoNT-A on fasciitis pain, with the primary outcome measure being the visual analog scale. We conducted statistical analyses using RevMan 5.4 software. RESULTS: Our final meta-analysis comprised 14 randomized controlled trials involving 537 participants, with 271 patients in the BoNT-A group and 266 patients in the control group. The overall effectiveness of BoNT-A in reducing fasciitis pain was significant, with a mean difference (MD) in visual analog scale score of -2.59 (95% confidence interval [CI], -3.36, -1.82); P < .00001; I2 = 88%. Subgroup analysis revealed that BoNT-A was particularly effective in treating plantar fasciitis (MD = -3.34 [95% CI, -4.08, -2.78]; P < .00001; I2 = 75%), lumbar back fasciitis (MD = -2.17 [95% CI, -3.82, -0.52]; P = .001; I2 = 93%), and neck and shoulder fasciitis (MD = -1.49 [95% CI, -2.76, -0.22]; P = .02; I2 = 61%). CONCLUSION: BoNT-A has a significant analgesic effect on fasciitis pain. Therefore, BoNT-A presents a promising alternative treatment option for fasciitis (PROSPERO 2022: CRD42022382805).

Comprehensive analysis of candidate signatures of long non-coding RNA LINC01116 and related protein-coding genes in patients with hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is a long-term malignancy that causes high morbidities and mortalities worldwide. Notably, long non-coding RNAs (LncRNAs) have been identified as candidate targets for malignancy treatments. METHODS: LncRNA LINC01116 and its Pearson-correlated genes (PCGs) were identified and analyzed in HCC patients. The diagnostic and prognostic value of the lncRNA was evaluated using data from The Cancer Genome Atlas (TCGA). Further, we explored the target drugs of LINC01116 for clinical application. Relationships between immune infiltration and PCGs, methylation and PCGs were explored. The diagnostic potentials were then validated by Oncomine cohorts. RESULTS: LINC01116 and the PCG OLFML2B are differentially and highly expressed in tumor tissues (both P ≤ 0.050). We found that LINC01116, TMSB15A, PLAU, OLFML2B, and MRC2 have diagnostic potentials (all AUC ≥ 0.700, all P ≤ 0.050) while LINC01116 and TMSB15A have prognostic significance (both adjusted P ≤ 0.050). LINC01116 was enriched in the vascular endothelial growth factor (VEGF) receptor signaling pathway, mesenchyme morphogenesis, etc. After that, candidate target drugs with potential clinical significance were identified: Thiamine, Cromolyn, Rilmenidine, Chlorhexidine, Sulindac_sulfone, Chloropyrazine, and Meprylcaine. Analysis of immune infiltration revealed that MRC2, OLFML2B, PLAU, and TMSB15A are negatively associated with the purity but positively associated with the specific cell types (all P < 0.050). Analysis of promoter methylation demonstrated that MRC2, OLFML2B, and PLAU have differential and high methylation levels in primary tumors (all P < 0.050). Validation results of the differential expressions and diagnostic potential of OLFML2B (Oncomine) were consistent with those obtained in the TCGA cohort (P < 0.050, AUC > 0.700). CONCLUSIONS: Differentially expressed LINC01116 could be a candidate diagnostic and an independent prognostic signature in HCC. Besides, its target drugs may work for HCC therapy via the VEGF receptor signaling pathway. Differentially expressed OLFML2B could be a diagnostic signature involved in HCC via immune infiltrates.

The growth-regulating factor PdbGRF1 positively regulates the salt stress response in Populus davidiana × P. bolleana.

Growth-regulating factor (GRF) is a transcription factor unique to plants that plays a crucial role in the growth, development and stress adaptation of plants. However, information on the GRFs related to salt stress in Populus davidiana × P. bolleana is lacking. In this study, we characterized the activity of PdbGRF1 in transgenic Populus davidiana × P. bolleana under salt stress. qRTPCR analyses showed that PdbGRF1 was highly expressed in young leaves and that the pattern of PdbGRF1 expression was significantly changed at most time points under salt stress, which suggests that PdbGRF1 expression may be related to the salt stress response. Moreover, PdbGRF1 overexpression enhanced tolerance to salt stress. A physiological parameter analysis showed that the overexpression of PdbGRF1 significantly decreased the contents of hydrogen peroxide (H2O2) and malondialdehyde (MDA) and increased the activities of antioxidant enzymes (SOD and POD) and the proline content. A molecular analysis showed that PdbGRF1 regulated the expression of PdbPOD17 and PdbAKT1 by binding to the DRE ('A/GCCGAC') in their respective promoters. Together, our results demonstrate that the binding of PdbGRF1 to DRE regulates genes related to stress tolerance and activates the associated physiological pathways, and these effects increase the ROS scavenging ability, reduce the degree of damage to the plasma membrane and ultimately enhance the salt stress response in Populus davidiana × P. bolleana.

RNAi Mediated Gene Silencing of Detoxification Related Genes in the Ectropis oblique.

Ectropis oblique is one of the main pests that feed on tea leaves. At present, the main control method is chemical control, but the long-term use of insecticides has been related to the development of insect resistance. One of the resistance mechanisms is the upregulation of relevant detoxification enzymes for defense. In this study, four genes with increased expression were screened from the gene sequences annotated from the transcriptome data of deltamethrin-treated larvae of E. oblique, which are acid phosphatase EoACP138, and cytochrome P450 EoCYP316, carboxylesterase EoCarE592 and acetylcholine esterase EoAchE989, respectively. The fourth instar larvae of E. oblique were stimulated by deltamethrin, chlorpyrifos and fenpropathrin respectively, and the expression levels of the genes were detected by qRT-PCR. The result showed that all four genes' expression had significantly increased under the stimulation of three insecticides. RNAi technology was used to silence the expression of genes of EoACP138, EoCYP316, EoCarE592 and EoAchE989 in the fourth instar larvae of E. oblique. The change in the expression levels of the above genes in the larvae treated with dsRNA and stimulated with pesticides was determined by qRT-PCR. The target genes have been effectively silenced after feeding on dsRNA and higher sensitivity with higher mortality to pesticides was observed in the larvae interfered with dsRNA. The above genes are related to the detoxification and metabolism of resistance of E. oblique, which lays a foundation for further study on the mechanism of insecticide resistance in E. oblique.

Transcriptomic Analysis of Cadmium Stressed Tamarix hispida Revealed Novel Transcripts and the Importance of Abscisic Acid Network.

Cadmium (Cd) pollution is widely detected in soil and has been recognized as a major environmental problem. Tamarix hispida is a woody halophyte, which can form natural forest on the desert and soil with 0.5 to 1% salt content, making it an ideal plant for the research on response to abiotic stresses. However, no systematic study has investigated the molecular mechanism of Cd tolerance in T. hispida. In the study, RNA-seq technique was applied to analyze the transcriptomic changes in T. hispida treated with 150 µmol L-1 CdCl2 for 24, 48, and 72 h compared with control. In total, 72,764 unigenes exhibited similar sequences in the Non-redundant nucleic acid database (NR database), while 36.3% of all these unigenes may be new transcripts. In addition, 6,778, 8,282, and 8,601 DEGs were detected at 24, 48, and 72 h, respectively. Functional annotation analysis indicated that many genes may be involved in Cd stress response, including ion bonding, signal transduction, stress sensing, hormone responses and ROS metabolism. A ThUGT gene from the abscisic acid (ABA) signaling pathway can enhance Cd resistance ability of T. hispida by regulating the production of ROS under Cd stress and inhibit absorption of Cd. The new transcriptome resources and data that we present in this study for T. hispida may facilitate investigation of molecular mechanisms governing Cd resistance.

Simultaneous quantification of 24 aldehydes and ketones in oysters (Crassostrea gigas) with different thermal processing procedures by HPLC-electrospray tandem mass spectrometry.

Aldehydes and ketones are secondary oxidation products resulting from lipid oxidation that occurs during food processing. These small molecule compounds not only have an impact on the quality, odor and flavor of food, but also play a role in the pathogenesis of many human diseases. In this study, a HPLC-MS/MS analytical method was developed and validated for the simultaneous determination of 24 aldehydes and ketones. The coefficients of determination (R2) for all aldehydes and ketones were higher than 0.9975 at the range of 0.2-2000 ng/mL. The recoveries were in the range 71.20-108.13% with RSD < 10%. The method was tested by analyzing lipids from oysters with different thermal processing (boiling, frying, roasting and air frying) procedures; the highest concentration for saturated aldehydes and ketones while the highest content of unsaturated aldehydes in boiling treatment. Meanwhile, fatty acid oxidative decomposition was in agreement with aldehydes and ketones formation. Moreover, principal component analysis, orthogonal partial least-squares discriminant analysis and variable importance in projection value showed that lipid oxidation is positively related to the formation of a variety of aldehydes and ketones.

Proteomics and bioinformatics reveal insights into neuroinflammation in the acute to subacute phases in rat models of spinal cord contusion injury.

Neuroinflammation is recognized as a hallmark of spinal cord injury (SCI). Although neuroinflammation is an important pathogenic factor that leads to secondary injuries after SCI, neuroprotective anti-inflammatory treatments remain ineffective in the management of SCI. Moreover, the molecular signatures involved in the pathophysiological changes that occur during the course of SCI remain ambiguous. The current study investigated the proteins and pathways involved in C5 spinal cord hemi-contusion injury using a rat model by means of 4-D label-free proteomic analysis. Furthermore, two Gene Expression Omnibus (GEO) transcriptomic datasets, Western blot assays, and immunofluorescent staining were used to validate the expression levels and localization of dysregulated proteins. The present study observed that the rat models of SCI were associated with the enrichment of proteins related to the complement and coagulation cascades, cholesterol metabolism, and lysosome pathway throughout the acute and subacute phases of injury. Intriguingly, the current study also observed that 75 genes were significantly altered in both the GEO datasets, including ANXA1, C1QC, CTSZ, GM2A, GPNMB, and PYCARD. Further temporal clustering analysis revealed that the continuously upregulated protein cluster was associated with immune response, lipid regulation, lysosome pathway, and myeloid cells. Additionally, five proteins were further validated by means of Western blot assays and the immunofluorescent staining showed that these proteins coexisted with the F4/80+ reactive microglia and infiltrating macrophages. In conclusion, the proteomic data pertaining to the current study indicate the notable proteins and pathways that may be novel therapeutic targets for the treatment of SCI.

Overexpression of ThMYB8 mediates salt stress tolerance by directly activating stress-responsive gene expression.

MYB transcription factors are important in abiotic stress responses; however, the detailed mechanisms are unclear. Tamarix hispida contains multiple MYB genes. The present study characterized T. hispida MYB8 (ThMYB8) during salt stress using transgenic T. hispida and Arabidopsis assays. ThMYB8 overexpression and ThMYB8 RNAi analysis demonstrated that ThMYB8 enhanced the salt stress tolerance. Transgenic Arabidopsis ectopic expression of ThMYB8 significantly increased root growth, fresh weight, and seed germination rate compared with that of the wild-type under salt stress. Physiological parameters analysis in T. hispida and Arabidopsis showed that ThMYB8 overexpressing plants had the lowest levels of O2, H2O2, cell death, malondialdehyde, and electrolyte leakage. Overexpression of ThMYB8 regulated Na+ and K+ concentrations in plant tissues while maintaining K+/Na+ homeostasis. Analysis using qRT-PCR and ChIP-PCR identified possible downstream ThMYB8-regulated genes. ThMYB8 regulated the expression of ThCYP450-2 (cytochrome p450-2), Thltk (leucine-rich repeat transmembrane protein kinase), and ThTIP (aquaporin TIP) by binding to the MBSI motif ('CAACTG') in their promoters. The results indicated that ThMYB8 enhanced salt stress tolerance in T. hispida by regulating gene expression related to the activation of stress-associated physiological changes, such as enhanced reactive oxygen species scavenging capability, maintaining K+/Na+ homeostasis, and decreasing the malondialdehyde content and lipid peroxidation cell membranes.

Clinical value of nedaplatin-based chemotherapy combined with radiotherapy for locoregional advanced nasopharyngeal carcinoma: a retrospective, propensity score-matched analysis.

Aims: This study aimed to investigate the clinical value of induction chemotherapy (IC) with docetaxel, 5-fluorouracil plus nedaplatin followed by concurrent chemoradiotherapy (CCRT) with nedaplatin for locoregional advanced nasopharyngeal carcinoma (NPC). Materials and Methods: In total, 269 patients diagnosed with locoregional advanced NPC between June 2012 and June 2017 were retrospectively included and divided into two groups: IC (docetaxel plus nedaplatin and 5-fluorouracil) followed by nedaplatin-based CCRT (TNF + N group, n = 146) and IC (docetaxel plus cisplatin and 5-fluorouracil) followed by cisplatin-based CCRT (TPF + P group, n = 123). The Kaplan-Meier method and Cox proportional hazards model were applied to analyse survival and prognosis. After propensity score-matched (PSM), 113 patients remained in each group. Toxicities were compared between the two groups using the Chi-square test or Fisher's exact test. Results: The overall survival (OS), progression-free survival (PFS), distant metastasis-free survival (DMFS), and locoregional relapse-free survival (LRRFS) rates of the TNF + N and TPF + P groups were 90.7% vs. 92.3% (P = 0.315), 78.9% vs. 79.4% (P = 0.715), 82.4% vs. 85.1% (P = 0.441) and 96.1% vs. 93.3% (P = 0.414), respectively, with no significant difference in 3-year survival outcome between the two groups, and this outcome was confirmed after using PSM analyses. In the PSM cohort, a significant higher frequency of grade 3/4 vomiting was observed in the TPF + P group compared to the TNF + N group (22.1% vs. 0%, P = 0.000). However, 15.9% of patients in the TNF + N group had grade 3/4 thrombocytopenia in comparison with 6.2% in the TPF + P group (P = 0.020). Conclusions: The TNF regimen followed by CCRT with nedaplatin is an alternative treatment strategy to the standard TPF regimen followed by CCRT with cisplatin for patients with locoregional advanced NPC.

Formation and disappearance of aldehydes during simulated gastrointestinal digestion of fried clams.

The formation and disappearance of aldehydes during simulated gastrointestinal digestion (SD) of fried clams was investigated in order to shed light on the underlying mechanism. Results from the thiobarbituric acid reactive substance (TBARS) and fluorometric assays using a specific aldehyde probe indicated that the SD (with lipase) of fried clams initially reduced (at the gastric stage), but subsequently increased (mainly at the intestinal stage) the contents of total aldehydes. Meanwhile, eight specific aldehydes including propanal, acrolein, trans-2-pentenal, hexanal, trans,trans-2,4-octadienal, trans,trans-2,4-decadienal, 4-hydroxy-hexenal and 4-hydroxy-nonenal in the digested meal were determined by using a high-performance liquid chromatography-tandem electrospray ionization mass spectrometry (HPLC-ESI-MS/MS) method. Results indicated that the changes in the trend of the contents of the eight aforementioned aldehydes were similar to those of total aldehydes during SD (with lipase) of fried clams. However, a similar SD process without lipase time-dependently reduced the contents of total and individual aldehydes. Moreover, lipid classes and free fatty acids (FFAs) in the digested meal were determined to reveal the degree of hydrolysis of lipids during the SD process. Results indicated that the SD (with lipase) of fried clams significantly hydrolyzed triacylglycerols (TAG) and polar lipids (PL) and produced FFAs, but the SD process without lipase resulted in negligible lipid hydrolysis. Thus, our results demonstrated a positive correlation between lipid hydrolysis and aldehyde generation during the SD of fried clams. Alternatively, unsaturated FFAs instead of TAG and PL could have served as the main precursors for aldehyde generation due to their high oxidative susceptibility.

Impact of different drying processes on the lipid deterioration and color characteristics of Penaeus vannamei.

BACKGROUND: Fresh shrimp (Penaeus vannamei) deteriorates easily and the drying process is an important processing method for prolonging the shelf life of shrimp. The traditional drying method is hot-air-drying (HD), which can cause some problems such as nutrient loss, discoloration and lipid oxidation. In recent years, freeze-drying (FD) has been popular for removing moisture from food at lower temperatures, maintaining the structure of raw materials, and improving storage stability of products. In the present study, the effects of HD and FD on lipid and color of P. vannamei and the mechanisms involved were investigated. RESULTS: FD caused less lipid oxidation compared to HD; consequently, FD-processed shrimps had lower levels of primary and secondary oxidation products, as well as acid value, and higher contents of triacylglycerol, phosphatidylcholine, phosphatidylethanolamine, eicosapentaenoic acid and docosahexaenoic acid compared to HD-processed samples. Lipase and lipoxygenase played a role in the oxidation and hydrolysis of lipids during drying process. FD-processed shrimps had lower yellowness value and chromatic aberrations but a higher whiteness value compared to HD-processed samples. Correlation analysis showed that lipid oxidation, astaxanthin degradation and the Maillard reaction contributed to the changes of color. Principal component analysis indicated that FD caused less deterioration in quality compared to HD. CONCLUSION: In the present study, FD is recommended for preserving shrimp color and lipid nutrition in terms of lipid oxidation control. © 2020 Society of Chemical Industry.

Advanced oxidation protein products increase TNF-α and IL-1ß expression in chondrocytes via NADPH oxidase 4 and accelerate cartilage degeneration in osteoarthritis progression.

Interleukin (IL)-1ß and tumor necrosis factor (TNF)-α, in particular, control the degeneration of articular cartilage, making them prime targets for osteoarthritis (OA) therapeutic strategies. Advanced oxidation protein products (AOPPs) are prevalent in numerous diseases. Our previous work demonstrates that intra-articular injections of AOPPs accelerate regression of cartilage in OA models. Whether AOPPs exist in the course of OA and their effects on TNF-α and IL-1ß expression in chondrocytes are still unclear. This study confirmed that AOPPs levels in human synovial fluid were positively associated with severity of OA. We also found AOPPs deposition in articular cartilage in anterior cruciate ligament transection (ACLT) induced rodent OA models. AOPPs increased expression of TNF-α and IL-1ß in chondrocytes in vitro, which was inhibited by pre-treatment with SB202190 (p38-MAPK inhibitor) or apocynin (NADPH oxidase inhibitor) or NOX4 knockdown by siRNAs. Subsequently, we further verified in vivo that exogenous injection of AOPPs in OA mice up-regulated expression of TNF-α and IL-1ß in cartilage, which was blocked by treatment with apocynin. In parallel, apocynin attenuated articular cartilage degeneration resulting in substantially lower OARSI scores. Specifically, apocynin reduced NOX4, p-P38, TNF-α and IL-1ß and increased collagen II and glycosaminoglycan (GAG). This study demonstrated that AOPPs increased expression of TNF-α and IL-1ß in chondrocytes via the NADPH oxidase4-dependent and p38-MAPK mediated pathway, and accelerated cartilage degeneration in OA progression. These findings suggest an endogenous pathogenic role of AOPPs in OA progression. Targeting AOPPs-triggered cellular mechanisms might be a promising therapeutic option for patients with OA.

Effects of temperature and heating time on the formation of aldehydes during the frying process of clam assessed by an HPLC-MS/MS method.

This study aimed at investigating the formation and accumulation of 16 reactive aldehydes in clam (Ruditapes philippinarum) during oil frying in both the tissue and the oil using an HPLC-ESI-MS/MS methodology. After processing, the accumulation of acrolein, crotonaldehyde, pentanal, trans-2-hexenal, hexanal, trans, trans-2,4-heptadienal, heptanal, nonanal, trans, trans-2,4-decadienal and 4-hydroxy-2-nonenal was most noticeable in both fried clam and frying oil. Most of the aldehyde species showed a time- and temperature-dependent manner of formation and accumulation during frying due to continuous oxidative degradation under conditions employed. However, several species of aldehyde such as acrolein and trans-2-pentenal slightly decreased at higher temperatures and/or longer frying times, which may be due to the imbalance toward disappearance of aldehydes resulting from their evaporation under the extreme conditions. Presence of natural polyphenols in bamboo leaves significantly prevented the formation of aldehydes in both fried clam and frying oil due to their antioxidant activity (P < 0.05).

Characterization of glycerophospholipid molecular species in muscles from three species of cephalopods by direct infusion-tandem mass spectrometry.

More than 200 molecular species of glycerophospholipids (GP) including glycerophosphocholine (GPC), glycerophosphoethanolamine (GPE), glycerophosphoserine (GPS), lysoglycerophosphocholine (LGPC), lysoglycerophosphoethanolamine (LGPE) and lysoglycerophosphoserine (LGPS), as well as 18 kinds of sphingomyelin (SM) were characterized by using a direct infusion-tandem mass (MS/MS) spectrometry method for lipids from the muscles of cephalopods Sepiella maindroni, Octopus ocellatus and Loligo chinensis for the first time. The majority of the GP molecular species contained long-chain omega-3 polyunsaturated fatty acids (n-3 LC-PUFA), especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Therefore, cephalopods can be a good possible source of dietary GP carrying n-3 LC-PUFA. The total lipids were composed of phospholipid (PL, 72.29-83.32 wt% of total lipids), cholesterol (12.70-23.60 wt% of total lipids), triacylglycerol (1.86-2.93 wt% of total lipids), diacylglycerol (0.15-1.09 wt% of total lipids), monoacylglycerol (0.06-0.18 wt% of total lipids) and free fatty acid (0.72-1.86 wt% of total lipids). For PL, phosphatidylcholine (44.47-62.30 mol%), phosphatidylethanolamine (22.57-39.08 mol%), phosphatidylserine (6.15-10.18 mol%), phosphatidylglycerol (0.68-3.11 mol%), phosphatidylinositol (2.41-7.15 mol%) and lysophosphatidylcholine (1.84-5.24 mol%) were detected. Furthermore, the total lipids from the muscles of cephalopods Sepiella maindroni, Octopus ocellatus and Loligo chinensis contained 41.80-50.02 mol% of saturated fatty acids, 11.53-21.54 mol% of monounsaturated fatty acids and 36.67-40.82 mol% of PUFA, whilst DHA (15.25-26.71 mol%) and EPA (6.29-16.57 mol%) were found to account for the majority of the PUFA. With these data presented, cephalopod muscle can be considered as a healthy food for humans.

Effects of natural phenolics on shelf life and lipid stability of freeze-dried scallop adductor muscle.

Effects of natural phenolics on the shelf life of dried scallop adductor muscle predicted by accelerated shelf life testing (ALST) combined with Arrhenius model were investigated. This allows the food industries to reliably and rapidly determine the shelf life of dried shellfish species treated with antioxidants. The shelf life of dried scallop adductor muscle treated with antioxidants of bamboo leaves (AOB) and tea polyphenols (TP) was more than 1.70-fold that of dried control scallop adductor muscle. Thus, the highly nutritional value of dried scallop adductor muscle, based on its lipid constituents, is maintained during storage. OXITEST method further confirmed the improvement of lipid stability of antioxidant treated dried scallop adductor muscle by protecting polyunsaturated fatty acids, especially eicosapentaenoic and docosahexaenoic acids, against autoxidation. Moreover, the natural phenolics employed effectively limited lipid oxidation by breaking the autoxidative chain reaction and/or inhibiting free radical formation in dried scallop adductor muscle during storage.

Lipid Profile and Glycerophospholipid Molecular Species in Two Species of Edible Razor Clams Sinonovacula constricta and Solen gouldi.

Total lipids were extracted from razor clams Sinonovacula constricta and Solen gouldi, and the molecular species of glycerophospholipid (Gpl) including choline glycerophospholipid (ChoGpl), ethanolamine glycerophospholipid (EtnGpl), serine glycerophospholipid (SerGpl), inositol glycerophospholipid (InsGpl), lysoChoGpl, lysoEtnGpl, and lysoSerGpl were characterized using a direct-infusion tandem mass spectrometric method for the first time. Meanwhile, the lipid class composition and phospholipid (PL) class composition as well as the fatty acid (FA) composition of total lipids, triacylglycerol (TAG), and PL were also investigated. About 238 and 235 molecular species were characterized, respectively, in Sinonovacula constricta and Solen gouldi. The majority of the dominant Gpl molecular species contained n-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA), especially docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). Also, razor clam lipids contained a high-proportioned PL (52.19-65.41% of total lipids) and PUFA (47.94-54.81 mol%). Furthermore, PL contained a higher proportion of PUFA (63.05-67.13 mol%), especially DHA (20.04-22.47 mol%) and EPA (16.27-21.46 mol%) than TAG (the corresponding values being 33.73-34.45, 11.95-12.27, and 8.13-0.8.99 mol%, respectively). Meanwhile, phosphatidylcholine (44.38-46.21 mol%) and phosphatidylethanolamine (38.84-39.95 mol%) were dominant among PL. In consideration of the high proportion of PUFA-enriched Gpl, razor clam plays a great role in promoting human health.