Multiparametric MRI combined with clinical factors to predict glypican-3 expression of hepatocellular carcinoma.

Objectives: The present study aims at establishing a noninvasive and reliable model for the preoperative prediction of glypican 3 (GPC3)-positive hepatocellular carcinoma (HCC) based on multiparametric magnetic resonance imaging (MRI) and clinical indicators. Methods: As a retrospective study, the subjects included 158 patients from two institutions with surgically-confirmed single HCC who underwent preoperative MRI between 2020 and 2022. The patients, 102 from institution I and 56 from institution II, were assigned to the training and the validation sets, respectively. The association of the clinic-radiological variables with the GPC3 expression was investigated through performing univariable and multivariable logistic regression (LR) analyses. The synthetic minority over-sampling technique (SMOTE) was used to balance the minority group (GPC3-negative HCCs) in the training set, and diagnostic performance was assessed by the area under the curve (AUC) and accuracy. Next, a prediction nomogram was developed and validated for patients with GPC3-positive HCC. The performance of the nomogram was evaluated through examining its calibration and clinical utility. Results: Based on the results obtained from multivariable analyses, alpha-fetoprotein levels > 20 ng/mL, 75th percentile ADC value < 1.48 ×103 mm2/s and R2* value ≥ 38.6 sec-1 were found to be the significant independent predictors of GPC3-positive HCC. The SMOTE-LR model based on three features achieved the best predictive performance in the training (AUC, 0.909; accuracy, 83.7%) and validation sets (AUC, 0.829; accuracy, 82.1%) with a good calibration performance and clinical usefulness. Conclusions: The nomogram combining multiparametric MRI and clinical indicators is found to have satisfactory predictive efficacy for preoperative prediction of GPC3-positive HCC. Accordingly, the proposed method can promote individualized risk stratification and further treatment decisions of HCC patients.

N-n-Butyl Haloperidol Iodide Ameliorates Oxidative Stress in Mitochondria Induced by Hypoxia/Reoxygenation through the Mitochondrial c-Jun N-Terminal Kinase/Sab/Src/Reactive Oxygen Species Pathway in H9c2 Cells.

Both c-Jun N-terminal kinase (JNK) and reactive oxygen species (ROS) play important roles in myocardial ischemia/reperfusion (I/R) injury. Our previous studies suggest that N-n-butyl haloperidol iodide (F2) exerts cardioprotection by reducing ROS production and JNK activation caused by I/R. In this study, we hypothesized that there is a JNK/Sab/Src/ROS pathway in the mitochondria in H9c2 cells following hypoxia/reoxygenation (H/R) that induces oxidative stress in the mitochondria and that F2 exerts mitochondrial protective effects during H/R injury by modulating this pathway. The results showed that H/R induced higher-level ROS in the cytoplasm on the one hand and JNK activation and translocation to the mitochondria by colocalization with Sab on the other. Moreover, H/R resulted in mitochondrial Src dephosphorylation, and subsequently, oxidative stress evidenced by the increase in ROS generation and oxidized cardiolipin in the mitochondrial membranes and by the decrease in mitochondrial superoxide dismutase activity and membrane potential. Furthermore, treatment with a JNK inhibitor or Sab small interfering RNA inhibited the mitochondrial translocation of p-JNK, decreased colocalization of p-JNK and Sab on the mitochondria, and reduced Src dephosphorylation and mitochondrial oxidative stress during H/R. In addition, Src dephosphorylation by inhibitor PP2 increased mitochondrial ROS production. F2, like inhibitors of the JNK/Sab/Src/ROS pathway, downregulated the H/R-induced mitochondrial translocation of p-JNK and the colocalization of p-JNK and Sab on the mitochondria, increased Src phosphorylation, and alleviated the above-mentioned mitochondrial oxidative stress. In conclusion, F2 could ameliorate H/R-associated oxidative stress in mitochondria in H9c2 cells through the mitochondrial JNK/Sab/Src/ROS pathway.

N-n-Butyl Haloperidol Iodide, a Derivative of the Anti-psychotic Haloperidol, Antagonizes Hypoxia/Reoxygenation Injury by Inhibiting an Egr-1/ROS Positive Feedback Loop in H9c2 Cells.

Early growth response-1 (Egr-1), a transcription factor which often underlies the molecular basis of myocardial ischemia/reperfusion (I/R) injury, and oxidative stress, is key to myocardial I/R injury. Silent information regulator of transcription 1(SIRT1) not only interacts with and is inhibited by Egr-1, but also downregulates reactive oxygen species (ROS) via the Forkhead box O1(FOXO1)/manganese superoxide dismutase (Mn-SOD) signaling pathway. N-n-butyl haloperidol iodide (F2), a new patented compound, protects the myocardium against myocardial I/R injury in various animal I/R models in vivo and various heart-derived cell hypoxia/reoxygenation (H/R) models in vitro. In addition, F2 can regulate the abnormal ROS/Egr-1 signaling pathway in cardiac microvascular endothelial cells (CMECs) and H9c2 cells after H/R. We studied whether there is an inverse Egr-1/ROS signaling pathway in H9c2 cells and whether the SIRT1/FOXO1/Mn-SOD signaling pathway mediates this. We verified a ROS/Egr-1 signaling loop in H9c2 cells during H/R and that F2 protects against myocardial H/R injury by affecting SIRT1-related signaling pathways. Knockdown of Egr-1, by siRNA interference, reduced ROS generation, and alleviated oxidative stress injury induced by H/R, as shown by upregulated mitochondrial membrane potential, increased glutathione peroxidase (GSH-px) and total SOD anti-oxidative enzyme activity, and downregulated MDA. Decreases in FOXO1 protein expression and Mn-SOD activity occurred after H/R, but could be blocked by Egr-1 siRNA. F2 treatment attenuated H/R-induced Egr-1 expression, ROS generation and other forms of oxidative stress injury such as MDA, and prevented H/R-induced decreases in FOXO1 and Mn-SOD activity. Nuclear co-localization between Egr-1 and SIRT1 was increased by H/R and decreased by either Egr-1 siRNA or F2. Therefore, our results suggest that Egr-1 inhibits the SIRT1/FOXO1/Mn-SOD antioxidant signaling pathway to increase ROS and perpetuate I/R injury. F2 inhibits induction of Egr-1 by H/R, thereby activating SIRT1/FOXO1/Mn-SOD antioxidant signaling and decreasing H/R-induced ROS, demonstrating an important mechanism by which F2 protects against myocardial H/R injury.

Chest circumference and birth weight are good predictors of lung function in preschool children from an e-waste recycling area.

The purpose of this study was to investigate the associations between birth weight, chest circumference, and lung function in preschool children from e-waste exposure area. A total of 206 preschool children from Guiyu (an e-waste recycling area) and Haojiang and Xiashan (the reference areas) in China were recruited and required to undergo physical examination, blood tests, and lung function tests during the study period. Birth outcome such as birth weight and birth height were obtained by questionnaire. Children living in the e-waste-exposed area have a lower birth weight, chest circumference, height, and lung function when compare to their peers from the reference areas (all p value <0.05). Both Spearman and partial correlation analyses showed that birth weight and chest circumference were positively correlated with lung function levels including forced vital capacity (FVC) and forced expiratory volume in 1 s (FEV1). After adjustment for the potential confounders in further linear regression analyses, birth weight, and chest circumference were positively associated with lung function levels, respectively. Taken together, birth weight and chest circumference may be good predictors for lung function levels in preschool children.

Considerable decrease of antibody titers against measles, mumps, and rubella in preschool children from an e-waste recycling area.

Data on vaccination effects in children chronically exposed to heavy metals are extremely scarce. This study aims to investigate the immune responsiveness to measles, mumps, and rubella (MMR) vaccination in children from an e-waste recycling area. 378 healthy children from Guiyu (exposed group) and Haojiang (reference group) were surveyed. Blood lead (Pb) levels were measured by graphite furnace atomic absorption. Titers of antibodies against MMR were quantified by ELISA. Blood Pb levels of children from the exposed group were significantly higher than those from the reference group (5.61µg/dL vs. 3.57µg/dL, p<0.001). In contrast, the antibody titers against MMR of the children from the exposed group were significantly lower than those from the reference group. The median titer of the anti-measles antibody of the exposed group was 669.64mIU/mL, with an interquartile range of 372.88-1068.42mIU/mL; this was decreased by nearly 40% compared to that of the reference group (median 1046.79mIU/mL, interquartile range 603.29-1733.10mIU/mL). For antibody titers against mumps, there was an about 45% decrease in the exposed group (median 272.24U/mL, interquartile range 95.19-590.16U/mL), compared to the reference group (median 491.78U/mL, interquartile range 183.38-945.96U/mL). In the case of rubella, the median titer of the antibody was also significantly lower in the exposed group (median 37.08IU/mL, interquartile range 17.67-66.66IU/mL) compared to the reference group (median 66.50IU/mL, interquartile range 25.32-105.59IU/mL); the decrease in this case was nearly 44%. The proportion of children whose antibody titers against MMR were below protective level in the exposed group was higher than it was in the reference group. The present study demonstrates that the immune responsiveness to routine vaccination was suppressed in children chronically exposed to lead. Thus, the vaccination strategies for these children living in an e-waste recycling area should be modified.

Biphasic Effects of Copper on Rat Learning and Memory in the Morris Water Maze.

Dysregulation of copper (Cu) metabolism interrupts neuron function, and subsequently results in neuron degeneration, necrosis, and gliocyte hyperplasia. To further explore the effects of hippocampal Cu concentration on learning and memory, Sprague-Dawley (SD) rats were given once-daily intraperitoneal injections of Copper(II) acetate (Cu(OAc)2) at doses of 0.2, 2, or 20mg/Kg over 5 days. Ultrasonic oscillation dialysis was used to determine the free Cu by graphite furnace atomic absorption spectrometry (GFAAS). Cu administration induced a dose-dependent increase in total hippocampal Cu. However, free hippocampal Cu was found to increase only at the lower concentration of Cu(OAc)2 (0.2 mg/Kg) but decrease at higher concentrations of Cu(OAc)2 (2 and 20 mg/Kg). Higher doses of Cu(OAc)2 (2-20mg/Kg) decreased superoxide dismutase-1 (SOD1) activity, increased both malondialdehyde (MDA) levels and the glutamate/γ-aminobutyric acid (Glu/GABA) ratio, and impaired spatial cognition. However, the lower dose of Cu(OAc)2 (0.2 mg/Kg) showed the opposite effects. This biphasic effect might be attributed to free hippocampal Cu levels and corresponding alterations of Glu/GABA ratio and SOD1 activity.

Chaotic time series prediction for prenatal exposure to polychlorinated biphenyls in umbilical cord blood using the least squares SEATR model.

Chaotic time series prediction based on nonlinear systems showed a superior performance in prediction field. We studied prenatal exposure to polychlorinated biphenyls (PCBs) by chaotic time series prediction using the least squares self-exciting threshold autoregressive (SEATR) model in umbilical cord blood in an electronic waste (e-waste) contaminated area. The specific prediction steps basing on the proposal methods for prenatal PCB exposure were put forward, and the proposed scheme's validity was further verified by numerical simulation experiments. Experiment results show: 1) seven kinds of PCB congeners negatively correlate with five different indices for birth status: newborn weight, height, gestational age, Apgar score and anogenital distance; 2) prenatal PCB exposed group at greater risks compared to the reference group; 3) PCBs increasingly accumulated with time in newborns; and 4) the possibility of newborns suffering from related diseases in the future was greater. The desirable numerical simulation experiments results demonstrated the feasibility of applying mathematical model in the environmental toxicology field.

Associations between maternal phenolic exposure and cord sex hormones in male newborns.

STUDY QUESTION: Are maternal urinary phenol concentrations associated with cord steroid hormone levels and anogenital distance (AGD) in male newborns? SUMMARY ANSWER: High maternal urinary Bisphenol A (BPA) levels are associated with decreases in cord testosterone levels and the ratio of testosterone to estradiol in male newborns, but there was no significant association with AGD. WHAT IS KNOWN ALREADY: Early life exposure to phenolic endocrine disrupting compounds (EDCs) is known to disrupt hormonal activities and affect reproductive development in males. However, studies on the health effects of prenatal human exposure are scarce. STUDY DESIGN, SIZE, DURATION: This was a cross-sectional study to investigate the association between maternal phenolic exposure and cord sex steroid hormones and AGD in male newborns. We recruited 100 mother-infant pairs from each of two hospitals, one in a polluted town (Guiyu) and the other in a cleaner town (Haojiang), from September 2010 to September 2011. PARTICIPANTS/MATERIALS, SETTING, METHODS: One hundred and seventy eight maternal urine samples and 137 cord blood samples were available for quantification, thus 137 complete records entered into the final analysis. Of them, 77 pairs were from Guiyu, and 60 were from Haojiang. The chemical concentrations were determined by solid phase extraction and gas chromatography-mass spectrometry (SPE-GC-MS), and cord sex hormones were detected by radioimmunoassay (RIA). Neonatal anthropometric parameters including AGD were measured. MAIN RESULTS AND THE ROLE OF CHANCE: Log2-transformed maternal urinary BPA concentration was negatively correlated with testosterone level and the ratio of testosterone to estradiol (T/E2) in male fetal cord blood after adjustment for potential confounders in linear regression models (ßadjusted = -31.09 (95% CI, -53.07 to -9.11) and ßadjusted = -0.08 (95% CI, -0.13 to -0.01), respectively). Moreover, compared with the lowest quartile group of BPA level, the highest group showed a significant decrease in testosterone level and T/E2 (ßadjusted = -179.84 (95% CI, -333.45 to -26.24) and ßadjusted = -0.37 (95% CI, -0.81 to 0.07), respectively). No significant associations between AGD or anogenital index (AGI, [AGI = AGD/birthweight (mm/kg)]) and phenolic EDCs or cord hormone levels were found. LIMITATIONS, REASONS FOR CAUTION: Results in the present study should be interpreted with caution because of its cross-sectional nature, small sample size and sampling time. WIDER IMPLICATIONS OF THE FINDINGS: Testosterone plays an important role in sex differentiation and normal development of the fetus and newborn, and the balance between testosterone and estradiol is thought an important mediator of prostate disease. Therefore, our findings may have important implications for human reproductive health. STUDY FUNDING/COMPETING INTERESTS: This work was supported by the National Natural Science Foundation of China (21377077) and Guangdong University Project for International Cooperation and Innovation Platform (2013gjhz0007). The authors declare they have no actual or potential competing financial interests.

N-n-butyl Haloperidol Iodide Protects against Hypoxia/Reoxygenation Injury in Cardiac Microvascular Endothelial Cells by Regulating the ROS/MAPK/Egr-1 Pathway.

Endothelium dysfunction induced by reactive oxygen species (ROS) is an important initial event at the onset of myocardial ischemia/reperfusion in which the Egr-1 transcription factor often serves as a master switch for various damage pathways following reperfusion injury. We hypothesized that an intracellular ROS/MAPK/Egr-1 signaling pathway is activated in cardiac microvascular endothelial cells (CMECs) following hypoxia/reoxygenation (H/R). ROS generation, by either H/R or the ROS donor xanthine oxidase-hypoxanthine (XO/HX) activated all three MAPKs (ERK1/2, JNK, p38), and induced Egr-1 expression and Egr-1 DNA-binding activity in CMECs, whereas ROS scavengers (EDA and NAC) had the opposite effect following H/R. Inhibitors of all three MAPKs individually inhibited induction of Egr-1 expression by H/R in CMECs. Moreover, N-n-butyl haloperidol (F2), previously shown to protect cardiomyocytes subjected to I/R, dose-dependently downregulated H/R-induced ROS generation, MAPK activation, and Egr-1 expression and activity in CMECs, whereas XO/HX and MAPK activators (EGF, anisomycin) antagonized the effects of F2. Inhibition of the ROS/MAPK/Egr-1 signaling pathway, by either F2, NAC, or inhibition of MAPK, increased CMEC viability and the GSH/GSSG ratio, and decreased Egr-1 nuclear translocation. These results show that the ROS/MAPK/Egr-1 signaling pathway mediates H/R injury in CMECs, and F2 blocks this pathway to protect against H/R injury and further alleviate myocardial I/R injury.

Effect of N-n-butyl haloperidol iodide on ROS/JNK/Egr-1 signaling in H9c2 cells after hypoxia/reoxygenation.

Reactive oxygen species (ROS)-induced oxidative stress in cells is an important pathophysiological process during myocardial ischemia/reperfusion (I/R) injury, and the transcription factor Egr-1 is a master switch for various damage pathways during reperfusion injury. An in vitro model of myocardial I/R injury and H9c2 cardiomyoblast cells hypoxia/reoxygenation (H/R) was used to assess whether there is abnormal intracellular ROS/JNK/Egr-1 signaling. We also assessed whether N-n-butyl haloperidol (F2), which exerts protective effects during myocardial I/R injury, can modulate this pathway. H/R induced ROS generation, JNK activation, and increased the expression of Egr-1 protein in H9c2 cells. The ROS scavengers edaravone (EDA) and N-acetyl-L-cysteine (NAC) reduced ROS level, downregulated JNK activation, and Egr-1 expression in H9c2 cells after H/R. The JNK inhibitor SP600125 inhibited Egr-1 overexpression in H9c2 cells caused by H/R. F2 could downregulate H/R-induced ROS level, JNK activation, and Egr-1 expression in H9c2 cells in a dose-dependent manner. The ROS donor hypoxanthine-xanthine oxidase (XO/HX) and the JNK activator ANISO antagonized the effects of F2. Therefore, H/R activates ROS/Egr-1 signaling pathway in H9c2 cells, and JNK activation plays an important role in this pathway. F2 regulates H/R-induced ROS/JNK/Egr-1 signaling, which might be an important mechanism by which it antagonizes myocardial I/R injury.

Association between blood erythrocyte lead concentrations and hemoglobin levels in preschool children.

Despite decades of intensive research, lead (Pb) toxicity still remains one of the most frequently investigated subjects in environmental health. Whole blood lead (BPb) is usually used to evaluate Pb exposure for both screening and clinical diagnosis. However, it is generally recognized that BPb is not a sensitive biomarker for Pb exposure in hematological studies. Considering hematocrit (HCT) variation in different situations, HCT-adjusted BPb or erythrocyte Pb (EPb) may be more relevant when evaluating the hematotoxicity of blood Pb. Data collected from 855 preschool children, 3-7 years of age, allowed us to examine the relationship between EPb and hemoglobin (Hb) levels. Multivariate linear regression was performed to determine the significance of EPb as predictor of Hb after covariate adjustment; then, mean differences of Hb levels between quartiles of EPb and BPb (1st quartile as reference) were determined using ANOVA followed by Student's t test. The dose-response curve between EPb and HCT was plotted using locally weighted scatterplot smoothing (LOWESS) method. A doubling of EPb was associated with a 2.44 g/L decrease in Hb level. Compared to the 1st quartile group of EPb, the 3rd and 4th quartile groups showed significant decreases in Hb levels (3.01 and 3.97 g/L, respectively). Compared to the 1st quartile group of BPb, the 2nd quartile group showed a decrease in Hb levels (0.63 g/L), while the 3rd and 4th quartile groups showed increases in Hb levels (0.78 and 1.45 g/L, respectively). Increased EPb levels are significantly associated with decreased Hb levels in preschool children. HCT must be taken into consideration in investigating the hematological effects of Pb. Compared to BPb, EPb or HCT-adjusted BPb appear as a more effective biomarker to interpret the hematotoxicity of lead. Furthermore, blood erythrocytes are not only a repository of Pb but also a primary target of its toxicity.

Polybrominated diphenyl ethers in human placenta associated with neonatal physiological development at a typical e-waste recycling area in China.

Our aim of this study was to characterize the exposure pattern of polybrominated diphenyl ethers(PBDEs) in human placenta and assess their potential effects on neonates. Placenta samples were obtained from a typical e-waste area in Guiyu and a reference area in Haojiang, China. The median ΣPBDE concentration was 32.25 ng/g lipid weight (lw) in placenta samples from Guiyu, and 5.13 ng/g lw from Haojiang. BDE-209 predominated in placenta samples, followed by BDE-28, -47, -99 -153, -183. Residence in Guiyu contributed the most to elevated PDBE levels. Neonatal physiological indices, including bodymass index (BMI), Apgar 1 score and head circumference, were reduced in Guiyu group. No significant difference was found in neonatal weight between the two groups, but neonatal body length in Guiyu was increased. Our data suggest prenatal exposure to PBDEs is high at the e-waste recycling area, and may lead to adverse physiological development in the fetus.

Chromium exposure among children from an electronic waste recycling town of China.

Guiyu is one of the most heavily chromium-polluted areas in China due to the numerous informal electronic waste (e-waste) recycling activities. A 3-year (2004, 2006, and 2008) independent cross-sectional study on blood chromium (BCr) levels of 711 children from Guiyu and a control area was investigated. Questionnaire completed by parents/guardians was used to assess the risk factors of chromium (Cr) exposure, while physical examination, for the year 2008 only, was used to evaluate the effects of long-term exposure to Cr on child physical development. Children living in Guiyu had significantly higher BCr levels compared with those living in Chendian at the same period from 2004 to 2008 (P < 0.001). The predominant risk factors related to elevated child BCr levels included the use of house as a family workshop, parent involved in e-waste recycling, and child residence in Guiyu. Children's weight and chest circumferences in group with high exposure to Cr (upper quartile) were higher than in the low-exposure group (P < 0.01), although the difference was less significant for boys between the two groups (P < 0.05). The results suggest that elevated child BCr in Guiyu due to informal e-waste recycling activities might be threatening the health of children, with implications on physical growth and development.

Assessment of association between the dopamine D2 receptor (DRD2) polymorphism and neurodevelopment of children exposed to lead.

The mechanism of lead (Pb) neurotoxicity has not been illustrated over the years. People pay more attention to dopaminergic neurotransmission, specifically dopamine receptor-2 (DRD2) Taq IA polymorphism, but no consensus has been reached. A total of 258 three-year-old children in Guiyu (exposed group) and Nanao (reference group), China were examined and their concentrations of blood lead (BPb) were determined. Cognitive and language scores of children were assessed using the Bayley Scales of Infant Development, third edition (BSID-III). Genotyping for the DRD2 polymorphism was carried out using a polymerase chain reaction (PCR) re-sequencing platform. The logistic stepwise regression analysis and stepwise regression analysis was used to explore associations among lead, neurodevelopment of children, and DRD2 Taq IA categories. Median values of Pb in Guiyu was higher than that of the reference group (11.30 ± 5.38 µg/dL vs. 5.77 ± 2.51 µg/dL, P < 0.001). Compared with the reference group, children from e-waste exposed area have lower cognitive scale scores (100 ± 25 vs 120 ± 20, P < 0.001) and lower language scale scores (99.87 ± 7.52 vs 111.39 ± 7.02, P < 0.001). The three kinds of genotype, A1/A1, A1/A2, and A2/A2, had no significant influences on BPb, cognitive scores and language scores (P > 0.05). Exposure of inhabitants, especially children to Pb from informal e-waste recycling activities might have contributed to higher levels of BPb and reduced cognitive and language scores observed in local children, however, the result obtained showed no significant association between DRD2 polymorphism and neurodevelopment of children exposed to lead.

Two ultrastructural distribution patterns of immunoglobulin G in human placenta and functional implications.

The placenta is known to protect the fetus from infection and maternal rejection. In a previous study, we demonstrated that placental trophoblasts can synthesize immunoglobulin G (IgG). In this study, we investigated the distribution of immunoglobulins (IgG, IgM, and IgA), IgG receptors (FcRn and FcgammaRIII), and complement proteins in placental trophoblasts at the ultrastructural level. In addition, we studied the mRNA expression of IgG1 heavy chain (IGHG1), recombination activating gene 1 (RAG1), RAG2, and activation-induced cytidine deaminase (AID) with nested RT-PCR in primary cultured trophoblasts. The mRNA transcripts of IGHG1, RAG1, RAG2, and AID were all identified in primary trophoblasts, further establishing the IgG-producing capacity of trophoblasts. At the ultrastructural level with colloidal gold-labeled antibodies, IgG was found to be distributed in two distinct locations in syncytiotrophoblasts. For one, it was colocalized with FcRn in endosome displaying low electron density, and for the other it was colocalized with complement C1q in medium-electron density irregular structures that have not been reported previously. This characteristic distribution suggests that IgG is likely processed through two molecular mechanisms in syncytiotrophoblasts: receptor-bound transportation across the syncytiotrophoblast and formation of immune complexes with locally produced IgG. The latter mechanism is probably aimed at neutralizing detrimental maternal anti-paternal major histocompatibility complex antibodies. Our findings support the hypothesis that placenta-produced IgG can selectively react with maternal anti-fetus antibodies and provide a mechanism of fetomaternal tolerance to protect the fetus from maternal immune rejection.

Actin cytoskeleton reorganization correlates with polarization of secretory vesicle and cell morphology in the degranulation of mast cell subtypes in human colon tissues.

Mast cells play a central role in the intestinal immune response. To investigate the relationship between degranulation, cell polarization and the reorganization of actin cytoskeleton of mast cells, we used fluorescence or gold labeling methods to identify different mast cell subtypes in human colon. The reorganization of filamentous actin was visualized and then the polarization of secretory vesicles, as well as cell surfaces, was analyzed by fluorescence microscopy and electron microscopy. Our results first showed a diversity of filamentous actin assembly or disassembly within the contacting cell membrane of different mast cell subtypes. The polarization and degranulation of secretory vesicles was not only accompanied with the assembly and disassembly of filamentous actin at the cell periphery, but also with changes of cell surface polarization. Our study provides an insight into the local membranous structures and suggested correlations of cytoskeleton arrangement with the polarization of secretory vesicles and cell surface configuration during mast cell degranulation.

Polybrominated diphenyl ethers in residential and agricultural soils from an electronic waste polluted region in South China: distribution, compositional profile, and sources.

A detailed investigation was conducted to understand the concentration, distribution, profile and possible source of polybrominated diphenyl ethers (PBDEs) in residential and agricultural soils from Guiyu, Shantou, China, one of the largest electronic waste (e-waste) recycling and dismantling areas in the world. Ten PBDEs were analyzed in 46 surface soil samples in terms of individual and total concentrations, together with soil organic matter concentrations. Much higher concentrations of the total PBDEs were predicted in the residential areas (more than 2000 ng g(-1)), exhibiting a clear urban source, while in the agricultural areas, concentrations were lower than 1500 ng g(-1). PBDE-209 was the most dominant congener among the study sites, indicating the prevalence of commercial deca-PBDE. However signature congeners from commercial octa-PBDE were also found. The total PBDE concentrations were significantly correlated with each individual PBDE. Principal component analysis indicated that PBDEs were mainly distributed in three groups according to the number of bromine atoms on the phenyl rings, and potential source. This study showed that the informal e-waste recycling has already introduced PBDEs into surrounding areas as pollutant which thus warrants an urgent investigation into the transport of PBDEs in the soil-plant system of agricultural areas.

Ultrastructural observations of human epidermal melanocytes cultured on polyethylene terephthalate film.

OBJECTIVE: To assess the application of polyethylene terephthalate film as a supporting material of cultured human epidermal melanocytes and to observe the ultrastructure of human epidermal melanocytes in vitro. METHODS: Human epidermal melanocytes were isolated from 7- to 14-years old children foreskins and were cultured in M254 culture medium containing human melanocyte growth supplement. Cultured melanocytes were purified via a differential trypsinization method. Purified melanocytes were cultured on a film and prepared for transmission electron microscopy. RESULTS: In comparing with the cellular supporting materials polyvinylidene chloride and cellophane, human epidermal melanocytes only attached and grew on polyethylene terephthalate film. Melanocytes grown on polyethylene terephthalate film maintained an intact shape, and retained special ultrastructures characteristic of melanocytes such as dendrites. CONCLUSIONS: Polyethylene terephthalate film proved to be an ideal cellular supporting material for the cultivation of human epidermal melanocytes. Ultrastructure of melanocytes showed melanosomes are transferred both from the tip and middle section of dendrites.

N-n-butyl haloperidol iodide ameliorates cardiomyocytes hypoxia/reoxygenation injury by extracellular calcium-dependent and -independent mechanisms.

N-n-butyl haloperidol iodide (F2) has been shown to antagonize myocardial ischemia/reperfusion injury by blocking calcium channels. This study explores the biological functions of ERK pathway in cardiomyocytes hypoxia/reoxygenation injury and clarifies the mechanisms by which F2 ameliorates cardiomyocytes hypoxia/reoxygenation injury through the extracellular-calcium-dependent and -independent ERK1/2-related pathways. In extracellularcalcium-containing hypoxia/reoxygenation cardiomyocytes, PKCα and ERK1/2 were activated, Egr-1 protein level and cTnI leakage increased, and cell viability decreased. The ERK1/2 inhibitors suppressed extracellular-calcium-containing-hypoxia/reoxygenation-induced Egr-1 overexpression and cardiomyocytes injury. PKCα inhibitor downregulated extracellularcalcium-containing-hypoxia/reoxygenation-induced increase in p-ERK1/2 and Egr-1 expression. F2 downregulated hypoxia/reoxygenation-induced elevation of p-PKCα, p-ERK1/2, and Egr-1 expression and inhibited cardiomyocytes damage. The ERK1/2 and PKCα activators antagonized F2's effects. In extracellular-calcium-free-hypoxia/reoxygenation cardiomyocytes, ERK1/2 was activated, LDH and cTnI leakage increased, and cell viability decreased. F2 and ERK1/2 inhibitors antagonized extracellular-calcium-free-hypoxia/reoxygenation-induced ERK1/2 activation and suppressed cardiomyocytes damage. The ERK1/2 activator antagonized F2's above effects. F2 had no effect on cardiomyocyte cAMP content or PKA and Egr-1 expression. Altogether, ERK activation in extracellular-calcium-containing and extracellular-calcium-free hypoxia/reoxygenation leads to cardiomyocytes damage. F2 may ameliorate cardiomyocytes hypoxia/reoxygenation injury by regulating the extracellular-calcium-dependent PKCα/ERK1/2/Egr-1 pathway and through the extracellular-calcium-independent ERK1/2 activation independently of the cAMP/PKA pathway or Egr-1 overexpression.

Effects of N-n-butyl haloperidol iodide on the rat myocardial sarcoplasmic reticulum Ca(2+)-ATPase during ischemia/reperfusion.

We have previously shown that N-n-butyl haloperidol iodide (F(2)), a newly synthesized compound, reduces ischemia/reperfusion (I/R) injury by preventing intracellular Ca(2+) overload through inhibiting L-type calcium channels and outward current of Na(+)/Ca(2+) exchanger. This study was to investigate the effects of F(2) on activity and protein expression of the rat myocardial sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) during I/R to discover other molecular mechanisms by which F(2) maintains intracellular Ca(2+) homeostasis. In an in vivo rat model of myocardial I/R achieved by occluding coronary artery for 30-60 min followed by 0-120 min reperfusion, treatment with F(2) (0.25, 0.5, 1, 2 and 4 mg/kg, respectively) dose-dependently inhibited the I/R-induced decrease in SERCA activity. However, neither different durations of I/R nor different doses of F(2) altered the expression levels of myocardial SERCA2a protein. These results indicate that F(2) exerts cardioprotective effects against I/R injury by inhibiting I/R-mediated decrease in SERCA activity by a mechanism independent of SERCA2a protein levels modulation.