Reprogramming macrophage by targeting VEGF and CD40 potentiates OX40 immunotherapy.

The low clinical response rate of checkpoint blockades, such as PD-1 and CTLA-4, highlighted the requirements of agonistic antibodies to boost optimal T cell responses. OX40, a co-stimulatory receptor on the T cells, plays a crucial role in promoting T cell survival and differentiation. However, the clinical efficacy of anti-OX40 agonistic antibodies was unimpressive. To explore the mechanism underlying the action of anti-OX40 agonists to improve the anti-tumor efficacy, we analyzed the dynamic changes of tumor-infiltrating immune cells at different days post-treatments using single-cell RNA-sequencing (scRNA-seq). In this study, we found that tumor-infiltrating regulatory T (Treg) cells were reduced after two rounds of anti-OX40 treatment, but the increase of infiltration and activation of CD8+ effector T cells, as well as M1 polarization in the tumor were only observed after three rounds of treatments. Moreover, our group first analyzed the antitumor effect of anti-OX40 treatments on regulating the macrophages and discovered the dynamic changes of vascular endothelial growth factor (VEGF) and CD40 signaling pathways on macrophages, indicating their possibility to being potential combination targets to improve the anti-OX40 agonists efficacy. The combination of VEGFR inhibitors or anti-CD40 agonist antibody with anti-OX40 agonists exhibited more remarkable inhibition of tumor growth. Therefore, the mechanism-driven combination of anti-OX40 agonists with VEGFR inhibitors or anti-CD40 agonists represented promising strategies.

Role of prognostic gene DKK1 in oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is one of the most common squamous cell carcinomas of the head and neck. The morbidity and mortality rates of OSCC have increased in recent years. However, the pathogenesis of this disease remains unknown. The present study aimed to identify predictive biomarkers and therapeutic targets for OSCC. Bioinformatics screening of differentially expressed genes in OSCC was performed based on data from The Cancer Genome Atlas and Gene Expression Omnibus databases. Dickkopf Wnt signaling pathway inhibitor 1 (DKK1) was identified to be associated with survival, tumor immunity and DNA repair in OSCC. Furthermore, the effects of DKK1 were evaluated by the knockdown of DKK1 in two OSCC cell lines. The proliferation, clonogenicity, migration and invasion of the cells were assessed in vitro using Cell Counting Kit-8, colony formation, wound healing and Transwell assays, respectively, and were found to be inhibited by DKK1 knockdown. The present study suggests that DKK1 may be used in the prognosis of patients with OSCC and that targeting DKK1 is a potential strategy for OSCC therapy.

Anorectal manometry after rectal surgery: a case report with rectal bleeding and perforation.

Background: Anorectal manometry (ARM) is primarily used to assess anorectal sensorimotor function. ARM provides comprehensive information about the anal sphincter and rectal sensation in patients with constipation or fecal incontinence. Generally, ARM can be applied to both adults and children. This study is the first to report a case of post-rectal resection ARM with constipation complicating rectal bleeding and perforation in a patient with no history of rectal cancer, aiming to provide guidelines for the treatment of patients who undergo ARM after rectal surgery. Case Description: A 58-year-old female patient with previous history of "hypertension and cerebral infarction" underwent "proctocolectomy" for "rectal prolapse" 11+ months ago and recovered well after the operation, but developed constipation. The patient was admitted to our hospital for anorectal manometry (ARM) for "constipation for 11+ months". 2+ hours after the examination, the patient developed blood in the stool and was hospitalized in the Department of Anorectal Surgery of our hospital. The anorectal manometry was performed 2+ hours after the patient developed blood in the stool. The patient was discharged after 30 days of hospitalization without any complications. During treatment, the patient suffered from rectal rupture and perforation. Following timely and aggressive surgery, the patient recovered and was discharged without any complications. Conclusions: ARM with balloon, as a method for detecting anorectal function, should only be performed cautiously after rectal surgery, especially among patients with comorbidities. If patients suffer from anorectal bleeding and perforation during ARM, prompt and aggressive surgical intervention is necessary. At present, there is little literature on ARM teaching courses. To improve the operation level of ARM and reduce the incidence of complications, we should extend understandings of ARM, develop a systematic management plan, and continuously summarize ARM-related experiences.

Hourly measurement of PM2.5-bound nonpolar organic compounds in Shanghai: Characteristics, sources and health risk assessment.

Fine particulate matter (PM2.5)-bound nonpolar organic compounds (NPOCs), including polycyclic aromatic hydrocarbons (PAHs) and alkanes, are commonly used as typical molecular markers for detailed source identification. Online thermal desorption aerosol gas chromatography-mass spectrometry (TAG) system can obtain ambient data with hourly resolution, which is of great importance for investigating the diurnal characteristics and refined source identification of NPOCs. From June to October 2020, hourly ambient aerosol samples were collected and analyzed to investigate the characteristics and sources of 14 PAHs and 15 alkanes (C21-C35) in PM2.5 using TAG at a suburban site of Baoshan district in Shanghai, China. The average concentration of summed PAHs and alkanes during the sampling period was 1.27 ± 1.4 ng/m3 and 8.87 ± 3.46 ng/m3, respectively, in which Benzo[b]fluoranthene (BbF), Benzo[ghi]perylene (BghiP) and Indeno[1,2,3-cd]pyrene (IcdP) are the dominant PAHs species, with n-Heptacosane (C27), n-Nonacosane (C29) and n-Hentriacontane (C31) being the most abundant n-alkane species. Carbon preference index (CPI) and carbon maximum (Cmax) number indicated that the sources of alkanes shifted from biogenic-oriented (such as plant wax) in the summer to anthropogenic-dominated (such as fossil fuels) in the autumn. Results from trajectory cluster analysis and potential source contribution function (PSCF) modeling showed that alkanes were mainly from the middle and lower reaches of the Yangtze River Plain including Anhui, Jiangxi, and Zhejiang provinces, while PAHs were mainly from northeastern China. Positive Matrix Factorization (PMF) model results indicated that gasoline (41.48%) and diesel (21.82%) were the two major sources of PM2.5-bound PAHs in summer and fall of 2020 in Shanghai, followed by coal consumption or catering (19.96%) and biomass burning (16.74%). Diurnal variation of PAHs sources resolved by PMF showed characteristic features consistent with the corresponding anthropogenic activities. For example, gasoline vehicle exhaust showed higher concentrations during traffic rush hours; while coal consumption or catering presented higher concentrations during lunch times from 10:00 to 12:00. In addition, the TAG data coupling with PMF also can be capable for source appointment of short-duration episodes. Health risk assessment showed that adult women were at greater lifetime cancer risk (ILCR) than people in other age groups, and people may subject to higher health risks at morning and night time. This work demonstrates that hourly NPOCs measured by TAG are uniquely specific on refined source identification and investigation into the characteristics of diurnal variations.

Rice G protein γ subunit qPE9-1 modulates root elongation for phosphorus uptake by involving 14-3-3 protein OsGF14b and plasma membrane H+ -ATPase.

Heterotrimeric G protein is involved in plant growth and development, while the role of rice (Oryza sativa) G protein γ subunit qPE9-1 in response to low-phosphorus (LP) conditions remains unclear. The gene expression of qPE9-1 was significantly induced in rice roots under LP conditions. Rice varieties carrying the qPE9-1 allele showed a stronger primary root response to LP than the varieties carrying the qpe9-1 allele (mutant of the qPE9-1 allele). Transgenic rice plants with the qPE9-1 allele had longer primary roots and higher P concentrations than those with the qpe9-1 allele under LP conditions. The plasma membrane (PM) H+ -ATPase was important for the qPE9-1-mediated response to LP. Furthermore, OsGF14b, a 14-3-3 protein that acts as a key component in activating PM H+ -ATPase for root elongation, is also involved in the qPE9-1 mediation. Moreover, the overexpression of OsGF14b in WYJ8 (carrying the qpe9-1 allele) partially increased primary root length under LP conditions. Experiments using R18 peptide (a 14-3-3 protein inhibitor) showed that qPE9-1 is important for primary root elongation and H+ efflux under LP conditions by involving the 14-3-3 protein. In addition, rhizosheath weight, total P content, and the rhizosheath soil Olsen-P concentration of qPE9-1 lines were higher than those of qpe9-1 lines under soil drying and LP conditions. These results suggest that the G protein γ subunit qPE9-1 in rice plants modulates root elongation for phosphorus uptake by involving the 14-3-3 protein OsGF14b and PM H+ -ATPase, which is required for rice P use.

Encapsulation of Astragaloside with Matrix Metalloproteinase-2-Responsive Hyaluronic Acid End-Conjugated Polyamidoamine Dendrimers Improves Wound Healing in Diabetes.

Impaired wound healing that occurs in diabetics can result in many life-threatening complications associated with excessive expression of matrix metalloproteinases (MMPs), which mediate the proteolysis of major matrix constituents. In this study, the dendrimer polyamidoamine (PAMAM) and the polysaccharide hyaluronic acid (HA) were connected through the substrate polypeptide (Gly-PLGLAG-Cys) of MMP-2 to obtain the MMP-2-responsive nanocarrier HA-pep-PAMAM. Insoluble astragaloside (ASI) was encapsulated in this nanocarrier to achieve controlled release at the site of intractable wounds. The HA-pep-PAMAM-ASI was successfully prepared with an average diameter of 142.3 ± 28.9 nm. Immunohistochemical staining of the skin revealed that the hard-to-heal wounds of diabetic mice showed stronger expression of MMP-2 than the wounds of normal mice. HA-pep-PAMAM-ASI achieved 73.9% release in the presence of MMP-2, but only 13.5% in PBS. A dose-dependent effect of H2O2 on the proliferation of BJ and HaCaT cells was observed, and HA-pep-PAMAM-ASI treatment had the best antioxidant capacity with MMP-2 pretreatment. HA-pep-PAMAM-ASI significantly increased GSH levels and reduced reactive oxygen species (ROS) levels to achieve antioxidant effects. The MMP-2-pretreated HA-pep-PAMAM-ASI group showed more improved cell proliferation and migration abilities. Compared with ASI group, the expression of all wound-repair-related genes in the group of HA-pep-PAMAM-ASI was significantly increased, and HA-pep-PAMAM-ASI showed a pronounced in vivo therapeutic effect. Therefore, our results revealed that enzyme-responsive MMP-2-loaded PAMAM nanoparticles could promote wound healing in diabetes and may be a promising biomaterial for treatment.

Hexabromocyclododecanes promoted autophagy through the PI3K/Akt/mTOR pathway in L02 cells.

As additive brominated flame retardants, hexabromocyclododecanes (HBCDs) are being widely used in diverse artificial materials and products, including thermal insulation building materials, housings of electronic equipment, and upholstery textiles. Toxicology studies have shown that HBCDs exposure are closely related to hepatotoxicity and liver diseases. The present study is designed to explore how HBCDs affect cell apoptosis and autophagy process in a human hepatocyte cell line (L02) and to reveal the underline molecular mechanisms. Firstly, HBCDs could elevate the apoptosis rate of L02 cells dose-dependently. Three apoptosis related proteins (apoptotic protease activating factor 1 (Apaf-1), cysteinyl aspartate specific proteinase 3 (caspase-3) and cysteinyl aspartate specific proteinase 9 (caspase-9)) were observed to be up-regulated using western blotting method. Autophagy process was also started by HBCDs in L02 cells as indicated by the increased expressions of LC3-phosphatidylethanolamine conjugate (LC3-II) and other autophagic protein markers (Beclin-1, autophagy related protein 3 (Atg3), autophagy related protein 5 (Atg5), autophagy related protein 7 (Atg7) and autophagy related protein 16L1(Atg16L1)). The results of the green fluorescent protein (GFP)-microtubule-associated protein 1 light chain 3 (LC3) intracellular localization and fluorescence intensity further evidenced the activation of autophagy in L02 cells after treated with HBCDs. In addition, phosphatidylinositide 3-kinases/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway was activated in L02 cells by HBCDs, suggested by the increased expressions of related proteins. The inhibitors of PI3K (LY294002), DNA-activated protein kinase catalytic subunit (DNA-PKcs) (NU7441), Akt (MK2206), and mTOR (KU0063794) could obviously reduce the autophagic proteins prompted by HBCDs. The fluorescence intensities of GFP-LC3 transfected L02 cells were also decreased significantly after the application of these inhibitors. These results indicated that PI3K/Akt/mTOR pathway was participated in regulating autophagy process promoted by HBCDs. In above, HBCDs could induce mitochondrial-dependent apoptosis and autophagy in L02 cells, which was modulated by PI3K/Akt/mTOR pathway.

Detrimental Role of miRNA-144-3p in Intracerebral Hemorrhage Induced Secondary Brain Injury is Mediated by Formyl Peptide Receptor 2 Downregulation Both In Vivo and In Vitro.

Although microRNA-144-3p (miRNA-144-3p) has been shown to suppress tumor proliferation and invasion, its function in intracerebral hemorrhage (ICH)-induced secondary brain injury (SBI) remains unclear. Thus, this study was designed to investigate the role of miRNA-144-3p in ICH. To accomplish this, we used adult male Sprague-Dawley rats to establish an in vivo ICH model by injecting autologous blood, while cultured primary rat cortical neurons were exposed to oxyhemoglobin (OxyHb) to mimic ICH in vitro. To examine the role of miRNA-144-3p in ICH-induced SBI, we used an miRNA-144-3p mimic and inhibitor both in vivo and in vitro. Following ICH induction, we found miRNA-144-3p expression to increase. Additionally, we predicted the formyl peptide receptor 2 (FPR2) to be a potential miRNA-144-3p target, which we validated experimentally, with FPR2 expression downregulated when miRNA-144-3p was upregulated. Furthermore, elevated miRNA-144-3p levels aggravated brain edema and neurobehavioral disorders and induced neuronal apoptosis via the downregulation of FPR2 both in vivo and in vitro. We suspected that these beneficial effects provided by FPR2 were associated with the PI3K/AKT pathway. We validated this finding by overexpressing FPR2 while inhibiting PI3K/AKT in vitro and in vivo. In conclusion, miRNA-144-3p aggravated ICH-induced SBI by targeting and downregulating FPR2, thereby contributing to neurological dysfunction and neural apoptosis via PI3K/AKT pathway activation. These findings suggest that inhibiting miRNA-144-3p may offer an effective approach to attenuating brain damage incurred after ICH and a potential therapy to improve ICH-induced SBI.

The Spermine Synthase OsSPMS1 Regulates Seed Germination, Grain Size, and Yield.

Polyamines, including putrescine, spermidine, and spermine, play essential roles in a wide variety of prokaryotic and eukaryotic organisms. Rice (Oryza sativa) contains four putative spermidine/spermine synthase (SPMS)-encoding genes (OsSPMS1, OsSPMS2, OsSPMS3, and OsACAULIS5), but none have been functionally characterized. In this study, we used a reverse genetic strategy to investigate the biological function of OsSPMS1 We generated several homozygous RNA interference (RNAi) and overexpression (OE) lines of OsSPMS1 Phenotypic analysis indicated that OsSPMS1 negatively regulates seed germination, grain size, and grain yield per plant. The ratio of spermine to spermidine was significantly lower in the RNAi lines and considerably higher in the OE lines than in the wild type, suggesting that OsSPMS1 may function as a SPMS. S-Adenosyl-l-methionine is a common precursor of polyamines and ethylene biosynthesis. The 1-aminocyclopropane-1-carboxylic acid (ACC) and ethylene contents in seeds increased significantly in RNAi lines and decreased in OE lines, respectively, compared with the wild type. Additionally, the reduced germination rates and growth defects of OE lines could be rescued with ACC treatment. These data suggest that OsSPMS1 affects ethylene synthesis and may regulate seed germination and plant growth by affecting the ACC and ethylene pathways. Most importantly, an OsSPMS1 knockout mutant showed an increase in grain yield per plant in a high-yield variety, Suken118, suggesting that OsSPMS1 is an important target for yield enhancement in rice.

Andrographolide ameliorates intracerebral hemorrhage induced secondary brain injury by inhibiting neuroinflammation induction.

Microglia activation and neuroinflammation play important roles in intracerebral hemorrhage (ICH)-induced secondary brain injury (SBI). In this study, we attempted to investigate the potential effects of Andrographolide (Andro) on ICH-induced SBI and the possible mechanisms behind these effects. Andro treatment effectively reduced neuronal cell death and degeneration and alleviated neurobehavioral disorders and brain edema in vivo. In an in vitro study, microglia activation-induced neuronal cell death was ameliorated by Andro treatment. In addition, microglia activation and neuroinflammation were induced by ICH, exhibiting elevated cytokine levels, which could be reversed with Andro treatment. The levels of TNF-α and IL-6 were significantly decreased after treatment with Andro, both in vivo and in vitro, due to the inhibition of nuclear transcription factor-κB (NF-κB) signaling pathway activation. Meanwhile, Andro decreased the levels of IL-1ß and LDH, as well as microglia pyroptosis induced by ICH by suppressing the assembly of the nucleotide-binding oligomerization domain like receptor protein 3 (NLRP3) inflammasome. In summary, this study reveals an anti-inflammatory effect of Andro and its potential mechanisms, and it shows that Andro is a potential candidate for improving ICH-induced SBI.

Arabidopsis plasma membrane H+-ATPase genes AHA2 and AHA7 have distinct and overlapping roles in the modulation of root tip H+ efflux in response to low-phosphorus stress.

Phosphorus deficiency in soil is one of the major limiting factors for plant growth. Plasma membrane H+-ATPase (PM H+-ATPase) plays an important role in the plant response to low-phosphorus stress (LP). However, few details are known regarding the action of PM H+-ATPase in mediating root proton (H+) flux and root growth under LP. In this study, we investigated the involvement and function of different Arabidopsis PM H+-ATPase genes in root H+ flux in response to LP. First, we examined the expressions of all Arabidopsis PM H+-ATPase gene family members (AHA1-AHA11) under LP. Expression of AHA2 and AHA7 in roots was enhanced under this condition. When the two genes were deficient in their respective Arabidopsis mutant plants, root growth and responses of the mutants to LP were highly inhibited compared with the wild-type plant. AHA2-deficient plants exhibited reduced primary root elongation and lower H+ efflux in the root elongation zone. AHA7-deficient plants exhibited reduced root hair density and lower H+ efflux in the root hair zone. The modulation of H+ efflux by AHA2 or AHA7 was affected by the action of 14-3-3 proteins and/or auxin regulatory pathways in the context of root growth and response to LP. Our results suggest that under LP conditions, AHA2 acts mainly to modulate primary root elongation by mediating H+ efflux in the root elongation zone, whereas AHA7 plays an important role in root hair formation by mediating H+ efflux in the root hair zone.

Role for RIP1 in mediating necroptosis in experimental intracerebral hemorrhage model both in vivo and in vitro.

Cell death is a hallmark of second brain injury after intracerebral hemorrhage (ICH); however, the mechanism still has not been fully illustrated. In this study, we explored whether necroptosis, a type of regulated necrosis, has an essential role in brain injury after ICH. We found that inhibiting receptor-interacting protein 1 (RIP1) - a core element of the necroptotic pathway - by a specific chemical inhibitor or genetic knockdown attenuated brain injury in a rat model of ICH. Furthermore, necroptosis of cultured neurons could be induced by conditioned medium from microglia stimulated with oxygen hemoglobin, and this effect could be inhibited by TNF-α inhibitor, indicating that TNF-α secreted from activated microglia is an important factor in inducing necroptosis of neurons. Undoubtedly, overexpression of RIP1 increased conditioned medium-induced necroptosis in vitro, but this effect was partially diminished in mutation of serine kinase phosphorylation site of RIP1, showing that phosphorylation of RIP1 is the essential molecular mechanism of necroptosis, which was activated in the in vitro model of ICH. Collectively, our investigation identified that necroptosis is an important mechanism of cell death in brain injury after ICH, and inhibition of necroptosis may be a potential therapeutic intervention of ICH.

Mutations Preventing Regulated Exon Skipping in MET Cause Osteofibrous Dysplasia.

The periosteum contributes to bone repair and maintenance of cortical bone mass. In contrast to the understanding of bone development within the epiphyseal growth plate, factors that regulate periosteal osteogenesis have not been studied as intensively. Osteofibrous dysplasia (OFD) is a congenital disorder of osteogenesis and is typically sporadic and characterized by radiolucent lesions affecting the cortical bone immediately under the periosteum of the tibia and fibula. We identified germline mutations in MET, encoding a receptor tyrosine kinase, that segregate with an autosomal-dominant form of OFD in three families and a mutation in a fourth affected subject from a simplex family and with bilateral disease. Mutations identified in all families with dominant inheritance and in the one simplex subject with bilateral disease abolished the splice inclusion of exon 14 in MET transcripts, which resulted in a MET receptor (MET(Δ14)) lacking a cytoplasmic juxtamembrane domain. Splice exclusion of this domain occurs during normal embryonic development, and forced induction of this exon-exclusion event retarded osteoblastic differentiation in vitro and inhibited bone-matrix mineralization. In an additional subject with unilateral OFD, we identified a somatic MET mutation, also affecting exon 14, that substituted a tyrosine residue critical for MET receptor turnover and, as in the case of the MET(Δ14) mutations, had a stabilizing effect on the mature protein. Taken together, these data show that aberrant MET regulation via the juxtamembrane domain subverts core MET receptor functions that regulate osteogenesis within cortical diaphyseal bone.

Synthesis and cytotoxic activity of 3, 4, 11-trihydroxyl modified derivatives of bergenin.

To synthesize a series of 3-, 4-, and/or 11-trihydroxy modified bergenin derivatives and evaluated their cytotoxic activity in vitro. The phenolic hydroxyl groups of bergenin were protected by benzyl groups with benzyl bromide. Treatment of dibenzyl bergenin with the corresponding acid in the presence of EDC·HCl and DMAP in CH2Cl2, followed by hydrogenation over Pd/C catalysts, afforded derivatives of bergenin esters. All of the target compounds were identified by IR, MS, and (1)H NMR. Twenty-six novel and three known derivatives of bergenin esters were synthesized. Their cytotoxicity values were evaluated by the MTT assay on the inhibition of DU-145 and BGC-823 cells in vitro. Several triply-substituted (3a, 4a, 5a, 6a, 7a) and doubly-substituted (8b, 9b) bergenin derivatives exhibited higher cytotoxic activity than bergenin. The result showed that the size of substituents and the lipophilicity of the bergenin esters displayed an important role on their cytotoxic activity.

Factors influencing on the bioaccessibility of polybrominated diphenyl ethers in size-specific dust from air conditioner filters.

Size-specific concentrations and bioaccessibility of polybrominated diphenyl ethers (PBDEs) in dust from air conditioner filters were measured, and the factors influencing the PBDE bioaccessibility were determined. Generally, the PBDE concentrations increased with decreasing dust particle size, and BDE209 (deca-BDE) was generally the predominant congener. The bioaccessibility ranged from 20.3% to 50.8% for tri- to hepta-BDEs, and from 5.1% to 13.9% for BDE209 in dust fractions of varied particle size. The bioaccessibility of most PBDE congeners decreased with increasing dust particle size. The way of being of PBDE (adsorbed to dust surface or incorporated into polymers) in dust significantly influenced the bioaccessibility. There was a significant negative correlation between the tri- to hepta-BDE bioaccessibility and organic matter (OM) contents in dust. Furthermore, tri- to hepta-BDE bioaccessibility increased with increasing polarity of OMs, while with decreasing aromaticity of OMs. The tri- to hepta-BDE bioaccessibility significantly positively correlated with the surface areas and pore volumes of dust. Using multiple linear regression analysis, it was found that the OM contents and pore volumes of dust were the most important factors to influence the tri- to hepta-BDE bioaccessibility and they could be used to estimate the bioaccessibility of tri- to hepta-BDEs according to the following equation: bioaccessibility (%)=45.05-0.49 × OM%+1.79 × pore volume. However, BDE209 bioaccessibility did not correlate to any of these factors.

Organochlorine pesticides in fish from Taihu Lake, China, and associated human health risk assessment.

Because contaminants and nutrients always coexist in fish, the risk from contaminants and the benefit from nutrients, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are also concomitant via fish consumption. To investigate the risk and benefit via fish consumption, concentrations of dichlorodiphenyltrichloroethane and its metabolites (DDTs), and hexachlorocyclohexanes (HCHs) in the whole- and portion-muscles of fish from Taihu Lake, China, were measured. Based on the contaminant data and nutrients from our previous study, and the associated risk and benefit via fish consumption were estimated. The concentrations of DDTs and HCHs in the whole-muscles ranged from 7.8 × 10² to 3.4×10³ pg g⁻¹ ww, and from 67.3 to 300 pg g⁻¹ ww, respectively. Of DDTs and HCHs measured, p,p'-DDE and ß-HCH were respectively the most abundant pesticides. The composition profiles of DDTs and HCHs suggested that the pesticides were mainly historical residues. The benefit-risk quotient (BRQ) of EPA+DHA vs. POPs (persistent organic pollutants including data of DDTs, HCHs, and those of polychlorinated biphenyls and polybrominated diphenyl ethers cited from our previous study) via consumption of fish from Taihu Lake was calculated. As a result, to achieve the recommended EPA+DHA intake of 250 mg d⁻¹ for a healthy adult, the consumption of most fish species from the lake can cause cancer and non-cancer risks. However, the fish consumption at the rates of 44.9 g d⁻¹ by Chinese would not lead to the risks for most of the species. The results also suggested that the risk of consuming silver carp was generally lower than other fish species, and those of dorsal muscles were lower than ventral and tail muscles.

Formation of fused-ring 2'-deoxycytidine adducts from 1-chloro-3-buten-2-one, an in vitro 1,3-butadiene metabolite, under in vitro physiological conditions.

1-Chloro-3-buten-2-one (CBO) is a potential metabolite of 1,3-butadiene (BD), a carcinogenic air pollutant. CBO is a bifunctional alkylating agent that readily reacts with glutathione (GSH) to form mono-GSH and di-GSH adducts. Recently, CBO and its precursor 1-chloro-2-hydroxy-3-butene (CHB) were found to be cytotoxic and genotoxic in human liver cells in culture with CBO being approximately 100-fold more potent than CHB. In the present study, CBO was shown to react readily with 2'-deoxycytidine (dC) under in vitro physiological conditions (pH 7.4, 37 °C) to form four dC adducts with the CBO moieties forming fused rings with the N3 and N(4) atoms of dC. The four products were structurally characterized as 2-hydroxy-2-hydroxymethyl-7-(2-deoxy-ß-d-erythro-pentofuranosyl)-1,2,3,4-tetrahydro-6-oxo-6H,7H-pyrimido[1,6-a]pyrimidin-5-ium (dC-1 and dC-2, a pair of diastereomers), 4-chloromethyl-4-hydroxy-7-(2-deoxy-ß-d-erythro-pentofuranosyl)-1,2,3,4-tetrahydro-6-oxo-6H,7H-pyrimido[1,6-a]pyrimidin-5-ium (dC-3), and 2-chloromethyl-2-hydroxy-7-(2-deoxy-ß-d-erythro-pentofuranosyl)-1,2,3,4-tetrahydro-6-oxo-6H,7H-pyrimido[1,6-a]pyrimidin-5-ium (dC-4). Interestingly, dC-1 and dC-2 were stable under our experimental conditions (pH 7.4, 37 °C, and 6 h) and existed in equilibrium as indicated by HPLC analysis, whereas dC-3 and dC-4 were labile with the half-lives being 3.0 ± 0.36 and 1.7 ± 0.06 h, respectively. Decomposition of dC-4 produced both dC-1 and dC-2, whereas acid hydrolysis of dC-1/dC-2 and dC-4 in 1 M HCl at 100 °C for 30 min yielded the deribosylated adducts dC-1H/dC-2H and dC-4H, respectively. Because fused-ring dC adducts of other chemicals are mutagenic, the characterized CBO-dC adducts could be mutagenic and play a role in the cytotoxicity and genotoxicity of CBO and its precursors, CHB and BD. The CBO-dC adducts may also be used as standards to characterize CBO-DNA adducts and to develop potential biomarkers for CBO formation in vivo.

Tissue-specific distribution of fatty acids, polychlorinated biphenyls and polybrominated diphenyl ethers in fish from Taihu Lake, China, and the benefit-risk assessment of their co-ingestion.

The fish tissues from four species collected from Taihu Lake, China, were analyzed including dorsal, ventral, and tail muscles, heart, liver, and kidney. The highest and lowest concentrations of fatty acids were respectively observed in livers and muscles. There were significant intraspecies and interspecies differences in the compositions of most fatty acids among muscle, heart, liver, and kidney. All the tissues were generally beneficial for consumption considering fatty acids. People mainly consume the muscle. Hence, the benefits from two polyunsaturated fatty acids, i.e., eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and risks from PCBs and PBDEs via fish consumption were evaluated by calculating the benefit-risk quotient (BFQ) for the intake of fish muscle containing EPA+DHA vs. PCBs or PBDEs. The BFQ values considering carcinogenic and noncarcinogenic effects for PCBs were ∼3000 and 10 times higher than those of PBDEs via fish consumption to achieve the recommended EPA+DHA intake of 250 mg d(-1), respectively. The results also suggested that the risk consuming the dorsal muscle was generally lower than the ventral and tail muscles.