Zinc oxide nanoparticles induces cell death and consequently leading to incomplete neural tube closure through oxidative stress during embryogenesis.

The implementation of Zinc oxide nanoparticles (ZnO NPs) raises concerns regarding their potential toxic effects on human health. Although more and more researches have confirmed the toxic effects of ZnO NPs, limited attention has been given to their impact on the early embryonic nervous system. This study aimed to explore the impact of exposure to ZnO NPs on early neurogenesis and explore its underlying mechanisms. We conducted experiments here to confirm the hypothesis that exposure to ZnO NPs causes neural tube defects in early embryonic development. We first used mouse and chicken embryos to confirm that ZnO NPs and the Zn2+ they release are able to penetrate the placental barrier, influence fetal growth and result in incomplete neural tube closure. Using SH-SY5Y cells, we determined that ZnO NPs-induced incomplete neural tube closure was caused by activation of various cell death modes, including ferroptosis, apoptosis and autophagy. Moreover, dissolved Zn2+ played a role in triggering widespread cell death. ZnO NPs were accumulated within mitochondria after entering cells, damaging mitochondrial function and resulting in the over production of reactive oxygen species, ultimately inducing cellular oxidative stress. The N-acetylcysteine (NAC) exhibits significant efficacy in mitigating cellular oxidative stress, thereby alleviating the cytotoxicity and neurotoxicity brought about by ZnO NPs. These findings indicated that the exposure of ZnO NPs in early embryonic development can induce cell death through oxidative stress, resulting in a reduced number of cells involved in early neural tube closure and ultimately resulting in incomplete neural tube closure during embryo development. The findings of this study could raise public awareness regarding the potential risks associated with the exposure and use of ZnO NPs in early pregnancy.

[Construction and Optimization of CD19 Chimeric Antigen Receptor T Cells Derived from C57BL/6J Mice].

OBJECTIVE: To explore the stimulation conditions, optimal culture time and infection time of C57BL/6J mice CD3+ T cells in vitro, so as to improve the infection efficiency of CD19 chimeric antigen receptor T cells (mCD19 CAR-T). METHODS: Purified C57BL/6J mice CD3+ T cells were cultured in anti-CD3/CD28 coated, anti-CD3 coated+soluble anti-CD28 and anti-CD3 coated, respectively. The cells were stimulated in above three conditions for 12 h and 24 h, following with 24 h, 48 h and 72 h incubation and then the number of cell clones was recorded. C57BL/6J mice CD3+ T cells were stimulated for 12 h, 24 h, and 36 h under the above three conditions, then interleukin (IL)-2 (100 U/ml) was added. The number of cell clones was recorded under microscope at 24 h, 48 h, and 72 h of culture. After 24 h of stimulation, CD3+ T cells derived from C57BL/6J mice were infected with retrovirus for 48 h to establish mCD19 CAR-T cells, and the percentage of GFP+ CAR-T cells was detected by flow cytometry. RESULTS: The infection efficiency of mCD19 CAR-T cells derived from C57BL/6J mice was only 5.23% under the optimized conditions of mCD19 CAR-T cells derived from BALB/c mice. The number of clones of C57BL/6J mice CD3+ T cells was the highest in anti-CD3 coated+soluble anti-CD28 group after stimulated for 24 h and followed cultured for 48 h. After 24 hours of stimulation under the above conditions and 48 hours of culture with IL-2, the number of T cell proliferating clones in the anti-CD3 coated+soluble anti-CD28 group was significantly increased compared with the same group without IL-2, and the infection efficiency of CAR-T cells in this group reached 17.63%±4.17%. CONCLUSION: The optimal conditions for constructing CAR-T cells from C57BL/6J mice CD3+ T cells are different from those of BABL/c mice. T cells stimulated by anti-CD3 coated+soluble anti-CD28+IL-2 can obtain mCD19 CAR-T cells with the highest efficiency after retrovirus infection.

Innovative explorations: unveiling the potential of organoids for investigating environmental pollutant exposure.

As the economy rapidly develops, chemicals are widely produced and used. This has exacerbated the problems associated with environmental pollution, raising the need for efficient toxicological evaluation techniques to investigate the toxic effects and mechanisms of toxicity of environmental pollutants. The progress in the techniques of cell culture in three dimensions has resulted in the creation of models that are more relevant in terms of biology and physiology. This enables researchers to study organ development, toxicology, and drug screening. Adult stem cells (ASCs) and induced pluripotent stem cells (iPSCs) can be obtained from various mammalian tissues, including cancerous and healthy tissues. Such stem cells exhibit a significant level of tissue memory and ability to self-assemble. When cultivated in 3D in vitro environments, the resulting organoids demonstrate a remarkable capacity to recapitulate the cellular composition and function of organs in vivo. Recently, many tumors' tissue-derived organoids have been widely used in research on tumor pathogenesis, drug development, precision medicine, and other fields, including those derived from colon cancer, cholangiocarcinoma, liver cancer, and gastric cancer. However, the application of organoid models for evaluating the toxicity of environmental pollutants is still in its infancy. This review introduces the characteristics of the toxicity responses of organoid models upon exposure to pollutants from the perspectives of organoid characteristics, tissue types, and their applications in toxicology; discusses the feasibility of using organoid models in evaluating the toxicity of pollutants; and provides a reference for future toxicological studies on environmental pollutants based on organoid models.

Film-forming polymer solutions containing cholesterol myristate and berberine mediate pressure ulcer repair via the Wnt/ß-catenin pathway.

Pressure ulcer (PU) is a worldwide problem that is difficult to address because of the related inflammatory response, local hypoxia, and repeated ischaemia/reperfusion, causing great suffering and financial burden to patients. Traditional Chinese medicine turtle plate powder can treat skin trauma, but its composition is complex and inconvenient to use. Here, we combined cholesterol myristate (S8) with berberine (BBR), with anti-inflammatory and antibacterial effects, as a drug and used hydroxypropyl methylcellulose and polyvinylpyrrolidone K30 as carriers to construct a novel film-forming polymeric solution (S8 + BBR FFPS), comprehensively study its reparative effect on PU and explore the potential mechanism in rat PU models. The results showed that S8 + BBR FFPS inhibits excessive inflammatory response, promotes re-epithelialization, and promotes hair follicle growth during the healing process of PU, which may be related to the activation of the Wnt/ß-catenin signalling pathway by S8 + BBR FFPS to mediate hair follicle stem cell proliferation and maintain skin homeostasis. Therefore, S8 + BBR FFPS may be a potential candidate for the treatment of chronic skin injury, and its association with the Wnt/ß-catenin signalling pathway may provide new ideas to guide the design of biomaterial-based wound dressings for chronic wound repair.

Insight into the adsorption behaviors and bioaccessibility of three altered microplastics through three types of advanced oxidation processes.

Advanced oxidation processes (AOPs) can significantly alter the structural properties, environmental behaviors and human exposure level of microplastics in aquatic environments. Three typical microplastics (Polyethylene (PE), polypropylene (PP), and polystyrene (PS)) and three AOPs (Heat-K2S2O8 (PDS), UV-H2O2, UV-peracetic acid (PAA)) were adopted to simulate the process when microplastics exposed to the sewage disposal system. 2-Nitrofluorene (2-NFlu) adsorption experiments found the equilibrium time decreased to 24 hours and the capacity increased up to 610 µg g-1, which means the adsorption efficiency has been greatly improved. The fitting results indicate the adsorption mechanism shifted from the partition dominant on pristine microplastic to the physical adsorption (pore filling) dominant. The alteration of specific surface area (21 to 152 m2 g-1), pore volume (0.003 to 0.148 cm3 g-1) and the particle size (123 to 16 µm) of microplastics after AOPs are implying the improvement for pore filling. Besides, the investigation of bioaccessibility is more complex, AOPs alter microplastic with more oxygen-containing functional groups and lower hydrophobicity detected by XPS and water contact angle, those modifications have increased the sorption concentration, especially in the human intestinal tract. Therefore, this indicates the actual exposure of organic compounds loaded in microplastic may be higher than in the pristine microplastic. This study can help to assess the human health risk of microplastic pollution in actual environments.

Correlation analysis between the in vivo bioavailability and in vitro bioaccessibility of nitro PAHs in soil: Application of simplified FOREhST in vitro methods based on the Chinese pharmacopoeia.

In this study, the relative bioavailability (RBA) of nitrated polycyclic aromatic hydrocarbons (NPAHs) in soil samples (n = 30) was assessed using an in vivo mouse model. Based on the correlation between the bioaccessibility data obtained from the Tenax improved traditional Fed ORganic Estimation human Simulation Test (FOREhST) in vitro method (TITF) and the bioavailability data obtained from in vivo experiments, the TITF method was further optimized and simplified by referring to the "Pharmacopoeia of the People's Republic of China: Volume IV, 2020" to adjust the formulation and parameters of the gastrointestinal fluid (GIF) in order to establish a simpler and lower cost in vitro method for the determination of the bioaccessibilities of NPAHs. The dose-accumulation relationship of the in vivo experiment showed that the linear dose-response was better in adipose tissue (R2 = 0.77-0.93), and the accumulation of NPAHs in adipose tissue was higher than that in kidney or liver tissue. Depending on the mouse adipose model, the NPAHs-RBA ranged from 1.88 % to 73.92 %, and a strongly significant negative relationship (R2 = 0.94, p < 0.05) was found between the NPAHs-RBA and Log Kow. The simplified experiment of the TITF showed that the composition of the GIF medium had a significant effect on the bioaccessibilities of NPAHs. The NPAH bioaccessibilities measured by the Tenax improved simplified FOREhST method (TISF) (9.0-36.5 %) were higher than that of the traditional FOREhST method (6.8-22.8 %) but significantly lower than that of the TITF method (16.8-55.2 %). With an increase in the bile concentration in the GIF (from 6 to 10 g/L), the bioaccessibilities of NPAHs increased from 9.0 to 36.5 % to 12.9-42.4 %. The accuracies of the four in vitro methods for predicting the bioavailabilities of NPAHs was in the following order: Tenax improved simplified FOREhST method with increased bile concentration (TITF-IB) (R2 = 0.54-0.87) ≈ TITF (R2 = 0.55-0.85) > TISF (R2 = 0.41-0.77) > FOREhST (R2 = 0.02-0.68). These results indicated that the simple in vitro method could also effectively predict the bioavailabilities of NPAHs.

Comparison of in vitro strategies for predicting Dichlorodiphenyltrichloroethane (DDT) and its metabolites bioavailability from soils.

In vitro strategies have widely been used to assess bioaccessibility of organic pollutants in soils. However, studies for comparing in vitro models with in vivo data are still limited. In this study, Dichlorodiphenyltrichloroethane (DDT) and its metabolites (called as DDTr) bioaccessibility in nine contaminated soils were measured using physiologically based extraction test (PBET), in vitro digestion model (IVD), and Deutsches Institut für Normung (DIN) with/without Tenax as an absorptive sink, and DDTr bioavailability was assessed using an in vivo mouse model. Whether or not Tenax was added, DDTr bioaccessibility significantly varied among three methods, suggesting that DDTr bioaccessibility depended on the in vitro method employed. Multiple linear regression analysis indicated that sink, intestinal incubation time and bile content are identified to be the dominant factors in controlling DDTr bioaccessibility. Comparison of in vitro and in vivo results demonstrated that DIN assay with Tenax (TI-DIN) provided the best prediction for DDTr bioavailability (r2 = 0.66, slope=0.78). After extending intestinal incubation time to 6 h or increasing bile content to 4.5 g/L (same to DIN assay) of the TI-PBET and TI-IVD assays, the in vivo-in vitro correlation will improved significantly, with r2 = 0.76 and slope= 1.4 for TI-PBET and r2 = 0.84 and slope= 1.9 for TI-IVD under 6 h intestinal incubation, and r2 = 0.59 and slope= 0.96 for TI-PBET and r2 = 0.51 and slope= 1.0 for TI-IVD under 4.5 g/L of bile content. The results suggest that it is essential to understand these key factors influencing bioaccessibility for the development of standardized in vitro methods, which helps to refine the risk assessment of human exposure to contaminants via soil ingestion.

Human nails as a valuable noninvasive alternative for estimating exposure to parabens.

Exposure of human to parabens (commonly used preservatives) is inevitable due to their extensively applied in numerous consumer products. Thus, a reliable noninvasive matrix reflecting long-term exposure to parabens is essential for human biomonitoring study. Human nails are potentially a valuable alternative for measuring intergrated exposure to parabens. In this work, we collected 100 paired nail and urine samples from university students in Nanjing, China, and measured simultaneously for six parent parabens and four metabolites. Methylparaben (MeP), ethylparaben (EtP), and propylparaben (PrP) were three predominant paraben analogue in both matrices, with the median concentrations being 12.9, 0.753, and 3.42 ng/mL in urine, and 1540, 154, and 961 ng/g in nail, respectively, while 4-hydroxybenzoic acid (4-HB) and 3,4-dihydroxybenzoic acid (3,4-DHB) were the most abundant metabolites (median values of 143 and 35.9 ng/mL, respectively) in urine. Gender-related analysis suggested that females exposed to more higher parabens than males. Significantly positive correlations were found between levels of MeP, PrP, EtP, and OH-MeP (r = 0.54-0.62, p < 0.01) in paired urine and nail samples. Our result here suggests that human nails, as an emerging biospecimen, are a potentially valuable biological matrix to evaluate human long-term exposure to parabens.

Insights into adsorption mechanisms of nitro polycyclic aromatic hydrocarbons on common microplastic particles: Experimental studies and modeling.

Nitro polycyclic aromatic hydrocarbons (NPAHs) and microplastics (MPs) are emerging contaminants that pose a threat to the aquatic ecosystem. Knowledge of the NPAHs and MPs interaction will help the understanding of their fate and risks in natural environment. Here, the adsorption behavior and mechanism of typical NPAHs on microplastics were investigated. The adsorption kinetic and isotherm data showed that the adsorption of NPAHs was controlled by chemical adsorption and hydrophobic partition, because of excellent fit of kinetic and isothermal equations (R2 > 0.9). The adsorption capacity (587-744 µg g-1) was largely dependent on the hydrophobicity of NPAHs. The experiment of environmental factors confirmed the important role of pollutant hydrophobicity, with 1-Npyr of the highest hydrophobicity having the greatest adsorption on MPs (adsorption rate >90%) and less affected by solution pH and ionic strength (changer <5%). In the mixture system, MPs displayed high adsorption capacity for each compound; Interestingly, because compounds with smaller size were easy to occupy the adsorption sites in the pores of MPs, the adsorption of 2-Nflu (724 µg g-1) was even greater than that of 9-Nant (713 µg g-1) and 1-Npyr (703 µg g-1). The model calculation of adsorption also shows that there is surface adsorption and hydrophobic distribution in the adsorption process. The findings provide new insights into the interactions of MPs with organic pollutants in complex environments.

The altering cellular components and function in tumor microenvironment during remissive and relapsed stages of anti-CD19 CAR T-cell treated lymphoma mice.

Anti-CD19 chimeric antigen receptor (CAR) T cells represent a highly promising strategy for B-cell malignancies. Despite the inspiring initial achievement, remission in a notable fraction of subjects is short-lived, and relapse remains a major challenge. Tumor microenvironment (TME) was proved to be aroused by CAR T cells; however, little is known about the dynamic characteristics of cellular components in TME especially during the different phases of disease after anti-CD19 CAR T-cell treatment. We took advantage of an immunocompetent model receiving syngeneic A20 lymphoma cells to dissect the changes in TME with or without CAR T-cell injection. We found that anti-CD19 CAR T-cell treatment attenuated the symptoms of lymphoma and significantly prolonged mice survival through eradicating systemic CD19+ cells. Increased myeloid subsets, including CD11c+ DCs and F4/80+ macrophages with higher MHC II and CD80 expression in bone marrow, spleen, and liver, were detected when mice reached remission after anti-CD19 CAR T treatment. Compared to mice without anti-CD19 CAR T administration, intrinsic T cells were triggered to produce more IFN-γ and TNF-α. However, some lymphoma mice relapsed by day 42 after therapy, which coincided with CAR T-cell recession, decreased myeloid cell activation and increased Treg cells. Elevated intrinsic T cells with high PD-1 and TIGIT exhaust signatures and attenuated cytotoxicity in TME were associated with the late-stage relapse of CAR T-cell treatment. In summary, the cellular compositions of TME as allies of CAR T cells may contribute to the anti-tumor efficacy at the initial stage, whereas anti-CD19 CAR T-cell disappearance and host response immunosuppression may work together to cause lymphoma relapse after an initial, near-complete elimination phase.

Effects of chemotherapy and immunotherapy on microbial diversity in TME and engineered bacterial-mediated tumor therapy.

Tumor microbiota is a group of microorganisms located in tumor tissues with rich diversity that can promote tumorigenesis and development, and different types of tumors have different tumor microbiotas, which has important implications for tumor research, detection, and clinical treatment. In this review, we examine the diversity of the tumor microbiota, discuss the impact of chemotherapy and immunotherapy on tumor microbiota diversity, and summarize recent advances in the use of genetically engineered bacteria for the treatment of tumors. In addition, we propose key questions that need to be further addressed by the tumor microbiota.

Bioaccessibility and bioavailability of NPAHs in soils using in vitro-in vivo assays: Comparison of laboratory and outdoor environmental aging effect.

Aging process is one of the most important factors that markedly reduces bioaccessibility and bioavailability (bioac-bioav) of organic contaminants. However, only few data on comparison of the effects of laboratory artificial aging (LAA) and outdoor environmental aging (OEA) processes on nitrated polycyclic aromatic hydrocarbons (NPAHs) bioac-bioav are available. In the current study, oral bioac-bioav of NPAHs in LAA and OEA soils (aging time intervals: 0, 45, 90, 120 and 150 d) were measured by in vitro traditional Fed ORganic Estimation human Simulation Test (FOREhST) and Tenax improved FOREhST (TI-FOREhST) methods, and in vivo mouse model. Tenax significantly increased the bioaccessibility of NPAHs in freshly spiked and aging soils from 0.3-40.9 % to 15.6-95.3 %, and 0.3-40.9 % to 1.0-84.5 %, respectively. Aging significantly reduced the NPAHs bioaccessibility (from 36.5 % to 10.7 %, and 12.1 % to 5.1 % as measured by FOREhST and TI-FOREhST, respectively) and bioavailability (from 27.7 % to 9.9 %, as measured by mouse model). The changes in bioac-bioav were mainly observed within the first 120 d of aging. The statistical analyses of NPAHs bioac-bioav showed no significant difference (p > 0.05) among the aging time intervals in LAA and OEA soils, which demonstrated that the LAA can relatively represent the OEA. Determination of TOC content in LAA and OEA soil can intuitively reflect whether the difference of NPAHs bioac-bioav between two aging treatment groups is significant. The mean bioaccessibility of NPAHs in soil measured by TI-FOREhST (mean 20.6 %) is closer to the bioavailability measured by mouse model (mean 19.4 %), indicating that Tenax improved in vitro method is more reliable than traditional methods, to predict the bioavailability of NPAHs.

Parabens in indoor dust from houses, university dormitories, and cosmetics stores in Nanjing, China: occurrence and human exposure.

Parabens are extensively used as preservatives in consumer products. The widespread exposure of human to parabens has been associated with adverse health effects. In this study, six parabens were measured in 100 indoor dust collected from homes, university dormitories, and cosmetics stores in Nanjing, China. Concentrations of sum of six parabens (∑6parabens) in dust from homes, university dormitories, and cosmetics stores ranged from 13.1 to 4.22 × 103, 102 to 3.03 × 103, and 7.02 × 103 to 3.41 × 104 ng/g, respectively. The median concentrations of ∑6parabens in dust from cosmetics stores (1.5 × 104 ng/g) were 1-2 orders of magnitude higher than those found in dust from homes (166 ng/g) and university dormitories (1.23 × 103 ng/g) (p < 0.01). Methyl-, ethyl-, and propyl-parabens were the predominant compounds found in dust samples, and the sum concentrations of three compounds accounted for 71.9-99.6%, 93.1-99.6%, and 94.7-99.6% of ∑6parabens in dust from homes, university dormitories, and cosmetics stores, respectively. Significant positive correlations were found between methyl- and propyl-parabens concentrations in three types of dust (r = 0.789-0.909), indicating their coexistence in many consumer products. The estimated daily intake (EDI) of ∑6parabens for adults via dust ingestion was highest for employees in cosmetics stores (median: 4.6 ng/kg bw/day), followed by university students (0.56-0.64 ng/kg bw/day), and adults in homes (0.075-0.087 ng/kg bw/day). The result provides a better understanding of human exposure to parabens in different indoor environments, and more studies are needed to further investigate the occurrence and potential health risks of parabens in dust from various microenvironments.

Morphology and cytochemical patterns of peripheral blood cells of tiger frog (Rana rugulosa).

Background: Tiger frog (Rana rugulosa) is a national second-class protected amphibian species in China with an important ecological and economic value. In recent years, due to excessive human hunting, pollution and habitat loss, the wild population of tiger frog has declined sharply. To protect wildlife resources, the artificial breeding of tiger frogs has rapidly developed in China. Diseases are increasing and spreading among tiger frogs due to the increasing scale of artificial farming. The blood examination is the most straightforward and less invasive technique to evaluate the animal health condition. Thus, it is essential to obtain the normal hematological indicators of tiger frogs. The objective of this study was to investigate the morphometry, microstructure and cytochemical patterns of peripheral blood cells in tiger frogs. Methods: The number of blood cells in tiger frogs was counted on a blood count board, and the cell sizes were measured by a micrometer under light microscope. The morphology and classification of blood cells were studied by Wright-Giemsa staining, and the cytochemical pateerns was investigated by various cytochemical staining including periodic acid-Schiff (PAS), Sudan black B (SBB), peroxidase (POX), alkaline phosphatase (AKP), acid phosphatase (ACP), chloroacetic acid AS-D naphthol esterase (CAE) and α-naphthol acetate esterase (ANAE) staining. Results: Besides erythrocytes and thrombocytes, five types of leukocytes were identified in tiger frogs: neutrophils, eosinophils, basophils, lymphocytes and monocytes. The mean erythrocyte, leukocyte and thrombocyte counts were 1.33 ± 0.15 million/mm3, 3.73 ± 0.04 × 104/mm3 and 1.7 ± 0.01 × 104/mm3, respectively. Small lymphocytes were the most abundant leukocytes, followed by large lymphocytes, Neutrophils, eosinophils and monocytes, basophils were the fewest. Eosinophils were strongly positive for PAS, positive for SBB, POX, ACP, CAE, ANAE, while weakly positive for AKP staining; basophils were strongly positive for PAS, ACP, positive for SBB, CAE, weakly positive for ANAE, negative for AKP, POX staining; neutrophils were strongly positive for ACP, SBB, positive for PAS, POX, weakly positive for AKP, CAE and ANAE staining; monocytes were positive for PAS, SBB, ANAE, weakly positive for ACP, AKP, POX, CAE staining; large lymphocytes and thrombocytes were positive for PAS, ACP, weakly positive for ANAE, while negative for SBB, POX, AKP, CAE; small lymphocytes were similar to large lymphocytes, except for strongly positive for PAS and ACP staining. Conclusions: The blood cell types and morphology of tiger frogs were generally similar to those of other amphibians, while their cytochemical patterns had some notable species specificity.Our study could enrich the knowledge of peripheral blood cell morphology and cytochemistry in amphibians, and provide baseline data for health condition evaluation and disease diagnosis of tiger frogs.

Developing and validating a chronic obstructive pulmonary disease quick screening questionnaire using statistical learning models.

BACKGROUND: Active targeted case-finding is a cost-effective way to identify individuals with high-risk for early diagnosis and interventions of chronic obstructive pulmonary disease (COPD). A precise and practical COPD screening instrument is needed in health care settings. METHODS: We created four statistical learning models to predict the risk of COPD using a multi-center randomized cross-sectional survey database (n = 5281). The minimal set of predictors and the best statistical learning model in identifying individuals with airway obstruction were selected to construct a new case-finding questionnaire. We validated its performance in a prospective cohort (n = 958) and compared it with three previously reported case-finding instruments. RESULTS: A set of seven predictors was selected from 643 variables, including age, morning productive cough, wheeze, years of smoking cessation, gender, job, and pack-year of smoking. In four statistical learning models, generalized additive model model had the highest area under curve (AUC) value both on the developing cross-sectional data set (AUC = 0.813) and the prospective validation data set (AUC = 0.880). Our questionnaire outperforms the other three tools on the cross-sectional validation data set. CONCLUSIONS: We developed a COPD case-finding questionnaire, which is an efficient and cost-effective tool for identifying high-risk population of COPD.

Ghrelin and ghrelin receptor (GHSR) in Chinese alligator, alligator sinensis: Molecular characterization, tissue distribution and mRNA expression changes during the active and hibernating periods.

The Chinese alligator (Alligator sinensis) is a freshwater crocodilian endemic to China. So far, the endocrine regulation of feeding and growth in Chinese alligator is poorly understood. In this study, the molecular structure and tissue expression profiles of ghrelin and its receptor GHSR in the Chinese alligator were characterized for the first time. The full-length cDNA of ghrelin was 1770 bp, including a 37 bp 5 '-UTR (untranslated region), a 435 bp ORF (open reading frame) and a 1298 bp 3 '-UTR. The ORF encodes a ghrelin precursor, which consists of 145 amino acid residues, including a signal peptide with 52 amino acid residues at the N-terminus, a mature peptide with 28 amino acid residues, and a possibly obestain at the C-terminus. The full-length cDNA of GHSR was 3961 bp, including a 5'-UTR of 375-bp, an ORF of 1059-bp and a 3' -UTR of 2527-bp. The ORF encodes a protein of 352 amino acid residues containing seven transmembrane domains, with multiple N glycosylation modification sites and conserved cysteine residue sites. The active core "GSSF" of Chinese alligator ghrelin was identical to that of mammals and birds, and the ghrelin binding site of GHSR was similar to that of mammals. The amino acid sequences of both ghrelin and GHSR share high identity with American alligator (Alligator mississippiensis) and birds. Ghrelin was highly expressed in cerebrum, mesencephalon, hypothalamus and multiple peripheral tissues, including lung, stomach and intestine, suggesting that it could play functions in paracrine and/or autocrine manners in addition to endocrine manner. GHSR expression level was higher in hypothalamus, epencephalon and medulla oblongata, and moderate in multiple peripheral tissues including lung, kindey, stomach and oviduct, implicating that ghrelin/GHSR system may participate in the regulation of energy balance, food intake, water and mineral balance, gastrointestinal motility, gastric acid secretion and reproduction. During hibernation, the expression of ghrelin and GHSR in the brain was significantly increased, while ghrelin was significantly decreased in heart, liver, lung, stomach, pancreas and ovary, and GHSR was significantly decreased in heart, liver, spleen, lung, kindey, stomach, ovary and oviduct. These temporal changes in ghrelin and GHSR expression could facilitate the physiological adaption to the hibernation of Chinese alligator. Our study could provide basic data for further studies on the regulation of feeding, physiological metabolism and reproduction of Chinese alligator, which could also be useful for the improvement of artificial breeding of this endangered species.

Ceramide synthase 3 affects invasion and metastasis of hepatocellular carcinoma via the SMAD6 gene. / ç¥žç»é °èƒºåˆæˆé ¶3通过SMAD6åŸºå› å½±å“è‚ç»†èƒžç™Œçš„ä¾µè¢­å’Œè½¬ç§».

OBJECTIVES: Patients with hepatocellular carcinoma (HCC) have poor prognosis due to lack of early diagnosis and effective treatment. Therefore, there is an urgent need to better understand the molecular mechanisms associated with HCC and to identify effective targets for early diagnosis and treatment. This study is to explore the expression and biological role of ceramide synthase 3 (CerS3) in HCC. METHODS: A total of 159 pairs of HCC tissues and adjacent non-tumor tissues were obtained from the patients underwent radical resection in Shenzhen People's Hospital, and the total RNA and proteins from HCC tissues and adjacent non-tumor tissues were obtained. The expression of CerS3 protein and mRNA in HCC was detected by immunohistochemistry, Western blotting and real-time PCR. In vitro experiments, Hep3B cells were divided into a control vector group and a CerS3 vector group, and the cells were transfected with retroviral vector containing control cDNA or CerS3 cDNA, respectively. HCCLM3 cells were divided into a normal control shRNA group and a CerS3 shRNA group, and the cells were transfected with lentiviral vectors containing normal control shRNA or CerS3 shRNA, respectively. MTT, EdU, Transwell and scratch method were used to detect cell proliferation, migration and invasion. RNA sequencing was performed to determine the downstream signal of CerS3. RESULTS: Compared with the corresponding adjacent tissues,the mRNA and protein levels of CerS3 were elevated in the HCC tissues, with significant difference (both P<0.05). The Univariate and multivariate analysis showed that the overall survival rate was significantly correlated with the presence of venous invasion (95% CI 1.8-9.2, P<0.01), TNM stage (95% CI 2.3-5.2, P<0.05), poor histological grade (95% CI 1.4-6.8, P<0.05), and CerS3 (95% CI 1.5-3.9, P<0.05). Furthermore, the high CerS3 expression levels in tumor tissues were significantly associated with shorter overall survival rates compared with the low CerS3 expression (P<0.05). Compared with the vector control group, the Hep3B cell viability, EdU positive cells, and migration and invasion cell numbers in the CerS3 vector group were significantly increased (all P<0.05). Compared with the shRNA normal control group, the HCCLM3 cell viability, EdU positive cells, and numbers of migrating and invasive cells in the CerS3 shRNA group were significantly lower (all P<0.05). The RNA sequencing confirmed that the small mothers against decapentaplegic family member 6 (SMAD6) gene as an oncogenic gene could promote the HCC metastasis. CONCLUSIONS: Clinically, the overexpression of CerS3 is closely related to poor clinical features and poor prognosis. Functionally, CerS3 participates in the proliferation, invasion and metastasis of liver cancer cells via activating SMAD6 gene.

A comparative study on adsorption behavior of iodinated X-ray contrast media iohexol and amidotrizoic acid by magnetic-activated carbon.

As persistent and ubiquitous contaminants in water, iodinated X-ray contrast media (ICM) pose a non-negligible risk to the environment and human health. In this study, we investigated the adsorption behavior of two typical ICM compounds, iohexol (IOH) and amidotrizoic acid (DTZ), on magnetic activated carbon. Theoretical investigations, using density functional theory, identified the molecule structures and calculated the molecular diameters of IOH (1.68 nm) and DTZ (1.16 nm), which revealed that ICM could be adsorbed by mesopores and larger micropores. Therefore, magnetic activated carbon with a porous structure was prepared by the co-precipitation method to investigate the adsorption mechanism of IOH and DTZ. MAC--5 (magnetic activated carbon with a theoretical iron oxide content of 37%) showed the best adsorption ability for both IOH and DTZ, with maximum adsorption capacities of 86.05 and 43.00 mg g-1, respectively. Adsorption kinetics and isotherm models were applied to explore the mechanisms involved, and the effects of solution pH, initial concentration, temperature, ionic strength, and natural organic matter were also investigated. The pore filling effect, π-π stacking, hydrogen bonding, and electrostatic interaction, were found to be the main adsorption mechanisms. The co-adsorption data showed that competition may occur in ICM coexisting environments. Interestingly, the used MAC--5 could be successfully regenerated and its adsorption efficiency did not decrease significantly after five cycles, indicating that it is a promising adsorbent for ICM. The results from this study provide some new insights for the treatment of water containing ICM.

Assessment of DDT and its Metabolites Bioaccessibility in Historically Contaminated Soils Using Unfed and Fed in Vitro Methods.

Bioaccessibility of hydrophobic organic contaminants (HOCs) from unintentional ingestion of soil is increasingly assessed with in vitro gastrointestinal models incorporating a sorption sink. In this study, the bioaccessibility of DDTs in contaminated soils (n = 11) was determined using "unfed" unified bioaccessibility method (UBM) and fed organic estimation human simulation test (FOREhST) with/without Tenax as an absorbent. By adding Tenax, the bioaccessibility of DDTs determined using UBM was significantly increased from 4.9-30.6% to 31.6-86.0%. In contrast, the bioaccessibility of DDTs determined using FOREhST without/with Tenax were similar with values of 20.0-60.9% vs 31.5-47.6%, implying that the influence of food components on the absorption efficiency of the sink should not be overlooked. Much high fraction of DDTs (bioaccessibility: 11.7-24.8%) remained in FOREhST supernatant after Tenax collection, suggesting that prediction of bioavailability through bioaccessibility obtained by absorbent needs to be treated with caution when bioaccessibility is determined using a "fed state" in vitro method.

Evaluating the adsorption performance of Tenax TA® in different containers: An isolation tool to study the bioaccessibility of nitro-PAHs in spiked soil.

The improved in vitro gastrointestinal simulation methods, with the addition of the adsorption sink, are considered as a promising tool for predicting the bioaccessibility of contaminants. However, the problem associated with the recovery of the adsorption sink from the complex matrix needs more understand. Although previous studies tried to solve this shortcoming by using the containers (a vessel to hold the adsorption sink), there is no systematic comparison study on the impact of containers on bioaccessibility till now, especially for nitro-polycyclic aromatic hydrocarbons (nitro-PAHs). In order to understand the problem, commonly used containers in previous studies (dialysis bags and stainless-steel screen) were selected and deployed in the Fed Organic Estimation Human Simulation Test (FOREhST) method to compare the effects of these containers on the bioaccessibility of nitro-PAHs desorbed from the five different types of soils into the gastrointestinal fluid (GIF). Results showed that in order to maintain a constant sorptive gradient for the high molecular weight (MW) nitro-PAHs, 0.25 g of Tenax TA® were required in FOREhST. Compared with Tenax TA® encapsulated in dialysis bag (Tenax-EDBG), the use of Tenax TA® encapsulated in dissolution basket (Tenax-EDBT) significantly increased the bioaccessibility of nitro-PAHs in the soil from 5.6-31.4% to 17.2-70.6%, due to the better diffusion performance. The bioaccessibility of nitro-PAHs by FOREhST extraction with Tenax-EDBT showed a significant negative correlation with soil total organic carbon (TOC), whereas a weak correlation with pH. This study provides the researchers with a more standardized in vitro method to quantify the bioaccessibility of PAHs and their derivatives in soil.