Recent advances of mechanosensitive genes in vascular endothelial cells for the formation and treatment of atherosclerosis.

Atherosclerotic cardiovascular disease and its complications are a high-incidence disease worldwide. Numerous studies have shown that blood flow shear has a huge impact on the function of vascular endothelial cells, and it plays an important role in gene regulation of pro-inflammatory, pro-thrombotic, pro-oxidative stress, and cell permeability. Many important endothelial cell mechanosensitive genes have been discovered, including KLK10, CCN gene family, NRP2, YAP, TAZ, HIF-1α, NF-κB, FOS, JUN, TFEB, KLF2/KLF4, NRF2, and ID1. Some of them have been intensively studied, whereas the relevant regulatory mechanism of other genes remains unclear. Focusing on these mechanosensitive genes will provide new strategies for therapeutic intervention in atherosclerotic vascular disease. Thus, this article reviews the mechanosensitive genes affecting vascular endothelial cells, including classical pathways and some newly screened genes, and summarizes the latest research progress on their roles in the pathogenesis of atherosclerosis to reveal effective therapeutic targets of drugs and provide new insights for anti-atherosclerosis.

Preoperative vascular heterogeneity based on dynamic susceptibility contrast MRI in predicting spatial pattern of locally recurrent high-grade gliomas.

OBJECTIVES: To investigate if spatial recurrence pattern is associated with patient prognosis, and whether MRI vascular habitats can predict spatial pattern. METHODS: In this retrospective study, 69 patients with locally recurrent high-grade gliomas (HGGs) were included. The cohort was divided into intra-resection cavity recurrence (ICR) and extra-resection cavity recurrence (ECR) patterns, according to the distance between the location of the recurrent tumor and the resection cavity or surgical region. Four vascular habitats, high angiogenic tumor, low angiogenic tumor, infiltrated peripheral edema, and vasogenic peripheral edema, were segmented and vascular heterogeneity parameters were analyzed. The survival and diagnostic performance under different spatial recurrence patterns were analyzed by Kaplan-Meier and ROC. A nomogram model was constructed by regression analysis and validated by bootstrapping technique. RESULTS: Progression-free survival (PFS) and overall survival (OS) were longer for ICR (n = 32) than those for ECR (n = 37) (median PFS: 8 vs. 5 months, median OS: 17 vs. 13 months, p < 0.05). MRI vascular habitat analyses showed ECR had higher median relative cerebral blood volume (rCBVmedian) at each habitat than ICR (all p < 0.01). The rCBVmedian at IPE had good diagnostic performance (AUC: 0.727, 95%CI: 0.607, 0.828). The AUC of the nomogram based on MRI vascular habitats and clinical factors was 0.834 (95%CI: 0.726, 0.913) and was confirmed as 0.833 (95%CI: 0.830, 0.836) by bootstrapping validation. CONCLUSIONS: The spatial pattern of locally recurrent HGGs is associated with prognosis. MRI vascular heterogeneity parameter could be used as a non-invasive imaging marker to predict spatial recurrence pattern. CLINICAL RELEVANCE STATEMENT: Vascular heterogeneity parameters based on MRI vascular habitat analyses can non-invasively predict the spatial patterns of locally recurrent high-grade gliomas, providing a new diagnostic basis for clinicians to develop the extent of surgical resection and postoperative radiotherapy planning. KEY POINTS: • Intra-resection cavity pattern was associated with longer progression-free survival and overall survival in locally recurrent high-grade gliomas. • Higher vascular heterogeneities in extra-resection cavity recurrence than in intra-resection cavity recurrence and the vascular heterogeneity parameters had good diagnostic performance in discriminating spatial recurrence pattern. • A nomogram model based on MRI vascular habitats and clinical factors had good performance in predicting spatial recurrence pattern.

Cellular gp96 upregulates AFP expression by blocking NR5A2 SUMOylation and ubiquitination in hepatocellular carcinoma.

Alpha-fetoprotein (AFP) is the most widely used biomarker for the diagnosis of hepatocellular carcinoma (HCC). However, a substantial proportion of HCC patients have either normal or marginally increased AFP levels in serum, and the underlying mechanisms are not fully understood. In the present study, we provided in vitro and in vivo evidence that heat shock protein gp96 promoted AFP expression at the transcriptional level in HCC. NR5A2 was identified as a key transcription factor for the AFP gene, and its stability was enhanced by gp96. A further mechanistic study by co-immunoprecipitation, GST pull-down, and molecular docking showed gp96 and the SUMO E3 ligase RanBP2 competitively binding to NR5A2 at the sites spanning from aa 507 to aa 539. The binding of gp96 inhibited SUMOylation, ubiquitination, and subsequent degradation of NR5A2. In addition, clinical analysis of HCC patients indicated that gp96 expression in tumors was positively correlated with serum AFP levels. Therefore, our study uncovered a novel mechanism that gp96 regulates the stability of its client proteins by directly affecting their SUMOylation and ubiquitination. These findings will help in designing more accurate AFP-based HCC diagnosis and progression monitoring approaches.

Effects of Culinary Procedures on Concentrations and Bioaccessibility of Cu, Zn, and As in Different Food Ingredients.

Although cooked diets are the primary sources for humans to absorb trace elements, there is limited data available on the concentrations and bioaccessibility of trace elements in cooked food ingredients. This work aims to evaluate the effects of culinary procedures on the concentrations and bioaccessibility of trace elements in common food ingredients. Twelve food species from the local market were treated with four culinary procedures (boiling, steaming, baking, and frying), then the bioaccessibility of copper (Cu), zinc (Zn), and arsenic (As) were evaluated using the in vitro digestion method. The subcellular distribution of these elements was also determined using the sequential fractionation method. The results show that culinary procedures decreased the retention rate of As during cooking (100% for raw and 65-89% for cooked ingredients) and the bioaccessibility of Cu and Zn during digestion (nearly 75% for raw and 49-65% for cooked ingredients), resulting in a reduction of the total bioaccessible fraction (TBF) of Cu, Zn, and As in food ingredients. The TBF of Cu, Zn, and As in all tested food ingredients followed the order: raw (76-80%) > steaming and baking (50-62%) > boiling and frying (41-50%). The effects of culinary procedures were associated with the subcellular distribution of trace elements. As was dominantly distributed in heat-stable proteins (51-71%), which were more likely to be lost during cooking. In comparison, Cu and Zn were mainly bound to the insoluble fraction and heat-denatured proteins (60-89% and 61-94% for Cu and Zn, respectively), which become less digestible in cooked ingredients. In conclusion, these results suggest that culinary procedures reduce the absorption of Cu, Zn, and As in various food ingredients, which should be considered in the coming studies related to nutrition and risk assessment of trace elements.

Effects of phosphate on the toxicity and bioaccumulation of arsenate in marine diatom Skeletonema costatum.

The effects of nutrient phosphate (P) at environmentally relevant levels on the toxicity of arsenic (As) in marine microalgae have been rarely known. In the present study, we explored the toxicity and bioaccumulation of As in a globally distributed diatom species Skeletonema costatum at different ambient P concentrations. The results showed that As toxicity was suppressed at elevated P concentrations. Intracellular As content ([As]intra) and the molar ratio of intracellular As to P ([As:P]) were negatively correlated with the ambient P concentrations. The trends of As bioaccumulation were substantially different between the relatively low (0, 0.5 and 1.5 µM) and high P concentrations (7.5 and 37.5 µM). Both [As]intra and [As:P] suggested that As bioaccumulation was a better factor to explain the As toxicity comparing to the ambient As concentration. The 4 h As uptake kinetics at different P concentrations followed Michaelis-Menten kinetic pattern. The maximum uptake rates (Vmax) decreased with the increase in P concentrations, and the half-saturation constants (Kd) remained constant except for that at extremely high P concentration (37.5 µM-P), suggesting the depression of P on As uptake was mainly due to the non-competitive effect. Overall, these results demonstrate that the P concentration in seawater is an important factor affecting As toxicity and bioaccumulation in the marine diatom. This study therefore helps us better understand the effects of eutrophication on the toxicity and biogeochemistry of As in the marine environment.

The strategy of arsenic metabolism in an arsenic-resistant bacterium Stenotrophomonas maltophilia SCSIOOM isolated from fish gut.

Bacteria are candidates for the biotransformation of environmental arsenic (As), while As metabolism in bacteria is not yet fully understood. In this study, we sequenced the genome of an As-resistant bacterium strain Stenotrophomonas maltophilia SCSIOOM isolated from the fish gut. After arsenate (As(V)) exposure, S. maltophilia transformed As(V) to organoarsenicals, along with the significant change of the expression of 40 genes, including the upregulation of arsH, arsRBC and betIBA. The heterogeneous expression of arsH and arsRBC increased As resistance of E. coli AW3110 by increasing As efflux and transformation. E. coli AW3110 (pET-betIBA) could transform inorganic As into dimethylarsinate (DMA) and nontoxic arsenobetaine (AsB), which suggested that AsB could be synthesized through the synthetic pathway of its analog-glycine betaine. In addition, the existence of arsRBC, betIBA and arsH reduced the reactive oxygen species (ROS) induced by As exposure. In total, these results demonstrated that S. maltophilia adopted an As metabolism strategy by reducing As accumulation and synthesizing less toxic As species. We first reported the production and potential synthetic pathway of AsB in bacteria, which improved our knowledge of As toxicology in microorganisms.

Promotor methylation status of MAPK4 is a novel epigenetic biomarker for prognosis of recurrence in patients with thymic epithelial tumors.

BACKGROUND: The prognosis of thymic epithelial tumors (TETs) currently relies on the commonly adopted WHO classification and Masaoka staging system, which cannot reflect the undefined biological behaviors limiting them as prognostic factors. METHODS: In this study, we first identified 40 genes and 179 genes, respectively that were epigenetically upregulated and silenced, corresponding to a total of 509 functionally methylated CpG sites between thymomas and thymic carcinomas by using the TCGA dataset. RESULTS: The methylation ß-values of cg20068620 in MAPK4 and cg18770944 in USP51 were significantly associated with recurrence-free survival (RFS). In the independent validation cohort, only WHO classification and methylation ß-values of cg20068620 in MAPK4 were independent prognostic factors for RFS in Chinese patients with TETs. A linear weighted model including these two factors was used to calculate the recurrence risk score (RRS). Time-dependent ROC curve analysis revealed that RRS was overwhelmingly superior to WHO classification for predicting 3-, 5-, and 10-year RFS and Masaoka stage for 3- and 5-year RFS. CONCLUSIONS: These results suggested that the methylation site cg20068620 in MAPK4 can improve the accuracy of the WHO classification alone regarding the prognostic value of TETs recurrence.

miR-146a Maintains Immune Tolerance of Kupffer Cells and Facilitates Hepatitis B Virus Persistence in Mice.

Kupffer cells (KCs), the largest tissue-resident macrophage population in the body, play a central role in maintaining a delicate balance between immune tolerance and immunity in the liver. However, the underlying molecular mechanism remains elusive. In this study, we show that KCs express high levels of miR-146a, which is under control of the PU.1 transcription factor. miR-146a deficiency promoted KCs differentiation toward a proinflammatory phenotype; conversely, miR-146a overexpression suppressed this phenotypic differentiation. We found that hepatitis B virus (HBV) persistence or HBV surface Ag treatment significantly upregulated miR-146a expression and thereby impaired polarization of KCs toward a proinflammatory phenotype. Furthermore, in an HBV carrier mouse model, KCs depletion by clodronate liposomes dramatically promoted HBV clearance and enhanced an HBV-specific hepatic CD8+ T cell and CD4+ T cell response. Consistent with this finding, miR-146a knockout mice cleared HBV faster and elicited a stronger adaptive antiviral immunity than wild-type mice. In vivo IL-12 blockade promoted HBV persistence and tempered the HBV-specific CTL response in the liver of miR-146a knockout mice. Taken together, our results identified miR-146a as a critical intrinsic regulator of an immunosuppressive phenotype in KCs under inflammatory stimuli, which may be beneficial in maintenance of liver homeostasis under physiological condition. Meanwhile, during HBV infection, miR-146a contributed to viral persistence by inhibiting KCs proinflammatory polarization, highlighting its potential as a therapeutic target in HBV infection.

Cadmium impairs zebrafish swim bladder development via ROS mediated inhibition of the Wnt / Hedgehog pathway.

The posterior swim bladder is an important organ in teleost fishes, that primarily maintains buoyancy and motility for swimming and survival. In this study, we examined the molecular mechanisms of the toxicity of cadmium (Cd) on the early development of the swim bladder in zebrafish. Embryonic Cd exposure resulted in the non-inflation of the swim bladder when the ambient Cd concentration was greater than or equal to 0.25 mg/L. Cd disturbed surfactant lipid distribution and inhibited the formation of all three tissue layers in the swim bladder. Additionally, excessive Cd down-regulated Wnt (fzd3, nkd1, fzd7 and axin2) and Hedgehog (ihh, shh, ptc1 and ptc2) signaling pathways. Conversely, Wnt signaling activation partially neutralized Cd-induced swim bladder developmental defects. Moreover, ROS scavenger reduced Glutathione (GSH) effectively recovered Cd induced defects in swim bladder and Wnt/Hedgehog signaling. Taken together, our results first revealed that Cd caused swim bladder developmental defects via ROS-mediated inhibition of the Wnt and Hedgehog pathways. These results herein provide important data for future toxicological studies and risk assessments of Cd.

Gut microbiota promote biotransformation and bioaccumulation of arsenic in tilapia.

Aquatic organisms such as fish can accumulate high levels of arsenic (As) and transform toxic inorganic As (iAs) to non-toxic arsenobetaine (AsB). Whether the gut microbiota are involved in the process of As accumulation and transformation in fish is unclear. Herein, we subjected tilapia (Oreochromis mossambicus) to antibiotic treatment for 19 d to remove the gut microbiota, followed by the dietary exposure to arsenate (As(V)) for 16 d. The antibiotic-treated fish accumulated significantly less total As and AsB levels in the intestine and muscle than the fish in the control group. The gut contents (mixture of microbiota, digestive fluid, and diet debris) in the control fish metabolized As(V) to arsenite (As(III)) and organoarsenicals in vitro, while those in the antibiotic-treated fish lost this ability. As(V) exposure significantly changed the fish gut microbiota community. Stenotrophomonas maltophilia was found to be the dominant species (>60% of total operational taxonomic unit (OTU) number) in the gut microbiota of As-treated fish. The isolated As-resistant strain S. maltophilia SCSIOOM owned a high capability to metabolize As(V) to As(III) and organoarsenicals. Overall, these results demonstrated that the gut microbiota, at least the As-resistant bacteria, were involved in As biotransformation pathways in fish.

Morphological and Hemodynamic Risk Factors for the Rupture of Proximal Anterior Cerebral Artery Aneurysms (A1 Segment).

OBJECTIVE: The treatment of unruptured small intracranial aneurysms remains controversial. A distinguishing characteristic of A1 segment aneurysms is that they tend to rupture when they are small, which may be related to their distinctive morphology and hemodynamics. Our study sought to investigate the rupture risk factors of A1 segment aneurysms by analyzing the clinical risk factors, morphology, and hemodynamic characteristics of A1 segment aneurysms. METHODS: We retrospectively enrolled 49 (23 ruptured, 26 unruptured) consecutive patients presenting to our institute with A1 segment aneurysms between January 2010 and March 2020. Independent risk factors associated with the rupture of A1 segment aneurysms were analyzed by multivariate regression analysis in the ruptured group and unruptured group. RESULTS: Clinical risk factors, including age, sex, hypertension, smoking history, and SAH family history revealed no difference between the ruptured and unruptured groups. The ruptured group presented a significantly larger size (Size, P = 0.007), aspect ratio (AR, P = 0.002), size ratio (SR, P = 0.001), bottleneck index (BN, P = 0.016), dome-to-neck ratio (DN, P = 0.001), and oscillatory shear index (OSI) (P = 0.001) than the unruptured group. The normalized wall shear stress (NWSS) of the ruptured aneurysms was lower than the unruptured group (P = 0.001). In the multivariate regression analysis, only SR (OR = 3.672, P = 0.003) and NWSS (OR = 0.474, P = 0.01) were independent risk factors in the A1 segment aneurysm rupture. CONCLUSION: A higher SR and lower NWSS revealed a close connection with the rupture of A1 segment aneurysms in our study, thus providing a reference for clinical decision-making in treating A1 segment unruptured aneurysms.

Induction of Foxp3 and activation of Tregs by HSP gp96 for treatment of autoimmune diseases.

Upregulation and stabilization of Foxp3 expression in Tregs are essential for regulating Treg function and immune homeostasis. In this study, gp96 immunization showed obvious therapeutic effects in a Lyn -/- mouse model of systemic lupus erythematosus. Moreover, gp96 alleviated the initiation and progression of MOG-induced experimental autoimmune encephalomyelitis. Immunization of gp96 increased Treg frequency, expansion, and suppressive function. Gene expression profiling identified the NF-κB family member p65 and c-Rel as the key transcription factors for enhanced Foxp3 expression in Treg by gp96. Mutant gp96 within its Toll-like receptor (TLR) binding domain, TLR2 knockout mice, and mice with cell-specific deletion of MyD88, were used to demonstrate that gp96 activated Tregs and induced Foxp3 expression via a TLR2-MyD88-mediated NF-κB signaling pathway. Taken together, these results show that gp96 immunization restricted antibody-induced and Th-induced autoimmune diseases by integrating Treg expansion and activation, indicating its potential clinical usefulness against autoimmune diseases.

TRPV1 activation inhibits phenotypic switching and oxidative stress in vascular smooth muscle cells by upregulating PPARα.

The proliferation and migration of vascular smooth muscle cells (VSMCs) is one of main reasons of vascular remodeling and is the pathogenesis of atherosclerosis and other vascular diseases. Transient receptor potential vanilloid 1 (TRPV1) is the specific receptor of capsaicin. TRPV1 has been previously reported to inhibit proliferation, migration and phenotypic switching, but the regulatory mechanisms and relevant signalling pathways are not clear. The aim of this study was to investigate the effects of capsaicin-activated TRPV1 on VSMC phenotypic switching. In this study, oxidized low density lipoprotein (ox-LDL) was used to induce the proliferation and migration of VSMCs. Our data showed that the VSMC proliferation induced by ox-LDL was dependent on the concentration of ox-LDL. Nevertheless, the data showed that capsaicin activated TRPV1 significantly decreased ox-LDL-induced superoxide anion generation. Phenotypic switching of VSMCs was inhibited by the activation of TRPV1. Furthermore, capsaicin decreased ox-LDL-induced superoxide anion generation by activating peroxisome proliferator activated receptor α (PPARα). TRPV1 inhibited VSMC phenotypic switching via upregulated expression of PPARα. It may be considered a useful target for the treatment of vascular remodeling.

A novel miR-7156-3p-HOXD13 axis modulates glioma progression by regulating tumor cell stemness.

Malignant glioma is the most common brain tumor in adults. Despite the great advances in anti-glioma treatments which have led to significant improvement in clinical outcomes, tumor recurrence remains the major cause of mortality. Increased cancer cell stemness and invasiveness are correlated with glioma progression. By searching the Cancer Genome Atlas, we showed that the expression of miR-7156-3p is significantly decreased in glioma tissues compared to the normal brain, and the decreased level of miR-7156-3p is closely correlated with glioma grade and patient survival. Clinical study consistently confirmed that miR-7156-3p is negatively correlated with glioma grade. Cell culture and animal experiments revealed that inhibition of miR-7156-3p effectively stimulates glioma cell stemness, invasion, and growth. In contrast, the augmentation of miR-7156-3p inhibits these phenotypes. Using Next-generation sequencing combined with target prediction approach, Homeobox D13 (HOXD13) is identified as the target gene of miR-7156-3p and further validated by luciferase reporter assay and cell transfection experiments. Additional in vitro and animal experiments demonstrated that miR-7156-3p regulates glioma cell stemness, invasion, and growth by mediating HOXD13. In conclusion, our findings provide new insight into the regulation of glioma stemness and invasiveness and may propose a potential strategy for anti-glioma treatment. Moreover, miR-7156-3p may serve as a candidate biomarker for predicting glioma progression in clinical practice.

A novel role of FKN/CX3CR1 in promoting osteogenic transformation of VSMCs and atherosclerotic calcification.

Fractalkine (FKN) and its specific receptor CX3CR1 play a critical role in the pathogenesis of atherosclerosis including recruitment of vascular cells and the development of inflammation. However, its contribution to regulating the development of atherosclerotic calcification has not been well documented. Osteogenic transformation of vascular smooth muscle cells (VSMCs) is critical in the development of calcification in atherosclerotic lesions. In this study, for the first time, we evaluated the effect of FKN/CX3CR1 on the progression of VSMCs calcification and defined molecular signaling that is operative in the FKN/CX3CR1-induced osteogenic transformation of VSMCs. We found that high-fat diet induced atherosclerotic calcification in vivo was markedly inhibited in the Apolipoprotein E (ApoE) and CX3CR1 deficient (ApoE-/-/CX3CR1-/-) mice compared with their control littermates. FKN and CX3CR1 were both expressed in VSMCs and up-regulated by oxidized low-density lipoprotein (ox-LDL). FKN/CX3CR1 promoted the expression of osteogenic markers, including osteopontin (OPN), bone morphogenetic protein (BMP)-2 and alkaline phosphatase (ALP) and decreased VSMCs markers, including smooth muscle (SM) α-actin and SM22-α in a dose-dependent manner. The essential role of FKN/CX3CR1 in VSMCs calcification was further confirmed by lentivirus-mediated knockdown or overexpression of CX3CR1 blocked or accelerated osteogenic transformation of VSMCs. This response was associated with reciprocal up- and down-regulation of osteogenic factor, runt-related transcription factor 2 (RUNX2), transcription factors in osteoclast differentiation, receptor activator of nuclear factor-κB (RANK), RANK ligand (RNAKL) and osteoprotegerin (OPG), respectively. Inhibition of FKN/CX3CR1-activated Jak2/Stat3 signaling by the Jak/Stat inhibitor AG490 blocked osteogenic transformation of VSMCs and RUNX2 induction concurrently. Taken together, our data uncovered novel roles of FKN/CX3CR1 in promoting VSMC osteogenic transformation and atherosclerotic calcification by activating RUNX2 through Jak2/Stat3 signaling pathway and suppressing OPG. Our findings suggest that targeting FKN/CX3CR1 may provide new strategies for the prevention and treatment of atherosclerotic calcification.

Occurrence and distribution of lipophilic phycotoxins in a subtropical bay of the South China Sea.

Lipophilic phycotoxins (LPs) pose significant threats to the health of marine mammals, birds, and human beings. The distribution and components of lipophilic phycotoxins contamination in subtropical area in the South China Sea are rarely known. This study systematically assessed the composition, concentration, and distribution of typical LPs in a typical subtropical bay, Daya Bay located in the South China Sea. Phytoplankton, seawater, suspended particulate matter, sediments, and shellfish samples were simultaneously collected from Daya Bay, and analyzed using liquid chromatography with tandem mass spectrometry. Okadaic acid, dinophysistoxins-1, pectenotoxins-2, yessotoxin and its derivate homo-yessotoxin, azaspiracid-2, 13-desmethyl spirolide C and gymnodimine were widely spread in multiple media in Daya Bay. Pectenotoxins-2 was the most widely distributed and highly concentrated toxin in the marine environments of Daya Bay. Toxin homo-yessotoxin was only detected in sediments and shellfish samples, and none of yessotoxin group components were found in phytoplankton and seawater, indicating that sediments were the major source of yessotoxin in shellfish. The study strongly demonstrated the lipophilic phycotoxins accumulated in shellfish are multisource, not only derived from toxigenic algae, but also from other marine media containing lipophilic phycotoxins. This study systematically distinguished multi-pathways of bioaccumulation of LPs in the marine shellfish.

Sediment as a Potential Pool for Lipophilic Marine Phycotoxins with the Case Study of Daya Bay of China.

Marine sediments can reserve many environmental pollutants. Lipophilic marine phycotoxins (LMPs) are natural toxic substances widespread in the marine environment; however, evidence of their existence in sediment is scarce. In the present study, in order to explore the occurrence and distribution characteristics of LMPs in sediment, surface sediment samples collected from a tropical area of Daya Bay (DYB) at different seasons, were analyzed using liquid chromatography with tandem mass spectrometry (LC-MS/MS). According to the results, up to six toxin compounds were detected in sediment samples from DYB, OA and DTX1 had the highest levels, followed by PTX2, homo-YTX, AZA2, and GYM. Although AZA2 and GYM were found in most of the sediment, OA, DTX1, homo-YTX, and PTX2 were the predominant toxin compounds, and PTX2 was the most ubiquitous toxin in sediment. The spatial distribution of LMP components in the sediment fluctuated with sampling times, partially according to the physical-chemical parameters of the sediment. There are likely several sources for LMPs existing in surface sediments, but it is difficult to determine contributions of a specific toxin-source in the sediment. Therefore, marine sediments may be a toxin reservoir for LMPs accumulation in benthic organisms via food chains.

Apoptosis induction effect of Apocynum venetum polyphenol on human U87 glioma cells via NF-κB pathway.

Aim: Apocynum venetum polyphenol (AVP) was used in in vitro glioma cells culture to prove the growth inhibitory effect of AVP on human U87 glioma cells via NF-κB pathway. Materials & methods: The MTT assay, DAPI morphology, quantitative PCR and western blot experiments were used for determination in vitro. Results & conclusion: AVP can also induce U87 cancer cells apoptosis illustrated by DAPI morphology. AVP could enhance the mRNA and protein expression of IκB-α, TNF-α, TRAIL, caspase-3 and caspase-9 in U87 cancer cells and reduce those of NF-κBp65, cIAP-1, cIAP-2, TGF-ß2, CyclinD1, VEGF and IL-8. After ammonium pyrrolidine dithiocarbamate (PDTC) treatment, the NF-κBp65 expression was reduced in U87 cells, and AVP could raise these effects. The results of HPLC indicate that AVP mainly contains six constituents. The growth inhibitory effects of AVP on U87 glioma cells are predominantly from these natural active constituents.

A peptide-based inhibitor of gp96 suppresses HBsAg expression and HBV replication by upregulation of p53.

In hepatitis B virus (HBV) infection, the virus produces redundant hepatitis B surface antigen (HBsAg) that plays a key role in driving T-cell tolerance and viral persistence. However, currently available anti-HBV agents have no direct effect on HBsAg transcription and protein expression. In this study, we designed a heat shock protein gp96 inhibitor p37 with the cell penetrating peptide PTD (protein transduction domain of trans-activator of transcription), which mediated p37 internalization into hepatocytes. PTD-p37 effectively suppressed HBsAg expression and viral replication both in vitro and in vivo. We further provide evidence that PTD-p37 suppressed HBV enhancer/promoter activity via p53 upregulation. Moreover, PTD-p37 had antiviral activity against a lamivudine-resistant HBV strain. Considering that suppression of HBsAg expression is a major goal for treatment of HBV infection, our results provide a basis for developing a new therapeutic approaches targeting host factors against viral expression.

The dynamic changes of arsenic bioaccumulation and antioxidant responses in the marine medaka Oryzias melastigma during chronic exposure.

Arsenic (As) is a highly toxic metalloid to aquatic organisms, but the effects of low-dose chronic inorganic As exposure on marine fish are still unclear. A 28-day study was conducted on chronic exposure of 100 µg/L inorganic As [As(III) and As(V)] in the marine medaka Oryzias melastigma to quantify the effects of chronic inorganic As exposure on its bioaccumulation, biotransformation, oxidative stress, and antioxidant enzymes response. During the exposure period, chronic inorganic As exposure had no significant effect on the total As bioaccumulation except at 7 d for As(V) treatment. Based on the toxicokinetic data, the low As bioaccumulation was caused by low dissolved uptake (ku), internal transfer (k12, k21) and high efflux (ke1). The organic As were the predominated species (77 - 80%) and remained constant, while the inorganic As increased 35% (from 0.26 µg/g to 0.35 µg/g) during the initial 4-d exposure and then recovered. The increased inorganic As could be covered by the high contribution of organic As especially arsenobetaine (AsB) to the total As accumulation. Coincidently, the malondialdehyde (MDA) levels followed similar trends as the inorganic As concentrations, indicating that inorganic As bioaccumulation induced oxidative stress at the initial stage. Both the contents of the nonenzymatic antioxidant (glutathione, GSH) and the activities of the enzymatic antioxidants (superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and glutathione-S-transferase (GST)) increased initially and then decreased as the inorganic As concentrations, thus lowering the hydrogen peroxide (H2O2) levels and displaying a typical antioxidant defense mechanism. In summary, this study elucidated that although the marine medaka had a limited ability to accumulate waterborne As, the increase in the inorganic As at the early stage of exposure still caused toxic effects, which could be ignored by constant total As concentrations. Therefore, the toxicity of As could be underestimated if only the total As instead of the inorganic As is monitored in marine fish.