Desmoglein 2 and desmocollin 2 depletions promote malignancy through distinct mechanisms in triple-negative and luminal breast cancer.

BACKGROUND: Aberrant expressions of desmoglein 2 (Dsg2) and desmocollin 2(Dsc2), the two most widely distributed desmosomal cadherins, have been found to play various roles in cancer in a context-dependent manner. Their specific roles on breast cancer (BC) and the potential mechanisms remain unclear. METHODS: The expressions of Dsg2 and Dsc2 in human BC tissues and cell lines were assessed by using bioinformatics analysis, immunohistochemistry and western blotting assays. Wound-healing and Transwell assays were performed to evaluate the cells' migration and invasion abilities. Plate colony-forming and MTT assays were used to examine the cells' capacity of proliferation. Mechanically, Dsg2 and Dsc2 knockdown-induced malignant behaviors were elucidated using western blotting assay as well as three inhibitors including MK2206 for AKT, PD98059 for ERK, and XAV-939 for ß-catenin. RESULTS: We found reduced expressions of Dsg2 and Dsc2 in human BC tissues and cell lines compared to normal counterparts. Furthermore, shRNA-mediated downregulation of Dsg2 and Dsc2 could significantly enhance cell proliferation, migration and invasion in triple-negative MDA-MB-231 and luminal MCF-7 BC cells. Mechanistically, EGFR activity was decreased but downstream AKT and ERK pathways were both activated maybe through other activated protein tyrosine kinases in shDsg2 and shDsc2 MDA-MB-231 cells since protein tyrosine kinases are key drivers of triple-negative BC survival. Additionally, AKT inhibitor treatment displayed much stronger capacity to abolish shDsg2 and shDsc2 induced progression compared to ERK inhibition, which was due to feedback activation of AKT pathway induced by ERK inhibition. In contrast, all of EGFR, AKT and ERK activities were attenuated, whereas ß-catenin was accumulated in shDsg2 and shDsc2 MCF-7 cells. These results indicate that EGFR-targeted therapy is not a good choice for BC patients with low Dsg2 or Dsc2 expression. Comparatively, AKT inhibitors may be more helpful to triple-negative BC patients with low Dsg2 or Dsc2 expression, while therapies targeting ß-catenin can be considered for luminal BC patients with low Dsg2 or Dsc2 expression. CONCLUSION: Our finding demonstrate that single knockdown of Dsg2 or Dsc2 could promote proliferation, motility and invasion in triple-negative MDA-MB-231 and luminal MCF-7 cells. Nevertheless, the underlying mechanisms were cellular context-specific and distinct.

Paracrine and epigenetic control of CAF-induced metastasis: the role of HOTAIR stimulated by TGF-ß1 secretion.

BACKGROUND: The communication between carcinoma associated fibroblasts (CAFs) and cancer cells facilitate tumor metastasis. In this study, we further underlying the epigenetic mechanisms of CAFs feed the cancer cells and the molecular mediators involved in these processes. METHODS: MCF-7 and MDA-MB-231 cells were treated with CAFs culture conditioned medium, respectively. Cytokine antibody array, enzyme-linked immunosorbent assay, western blotting and immunofluorescence were used to identify the key chemokines. Chromatin immunoprecipitation and luciferase reporter assay were performed to explore the transactivation of target LncRNA by CAFs. A series of in vitro assays was performed with RNAi-mediated knockdown to elucidate the function of LncRNA. An orthotopic mouse model of MDA-MB-231 was conducted to confirm the mechanism in vivo. RESULTS: Here we reported that TGF-ß1 was top one highest level of cytokine secreted by CAFs as revealed by cytokine antibody array. Paracrine TGF-ß1 was essential for CAFs induced EMT and metastasis in breast cancer cells, which is a crucial mediator of the interaction between stromal and cancer cells. CAF-CM significantly enhanced the HOTAIR expression to promote EMT, whereas treatment with small-molecule inhibitors of TGF-ß1 attenuated the activation of HOTAIR. Most importantly, SMAD2/3/4 directly bound the promoter site of HOTAIR, located between nucleotides -386 and -398, -440 and -452, suggesting that HOTAIR was a directly transcriptional target of SMAD2/3/4. Additionally, CAFs mediated EMT by targeting CDK5 signaling through H3K27 tri-methylation. Depletion of HOTAIR inhibited CAFs-induced tumor growth and lung metastasis in MDA-MB-231 orthotopic animal model. CONCLUSIONS: Our findings demonstrated that CAFs promoted the metastatic activity of breast cancer cells by activating the transcription of HOTAIR via TGF-ß1 secretion, supporting the pursuit of the TGF-ß1/HOTAIR axis as a target in breast cancer treatment.

Changes in microRNAs associated with Twist-1 and Bcl-2 overexpression identify signaling pathways.

Epithelial-mesenchymal transition (EMT) is regulated by multiple signal transduction pathways. Twist-1 is one of the most important transcription factors in these pathways. In a previous study, we found that Bcl-2 enhanced the role of Twist-1 in EMT. Coexpression of Twist-1 and Bcl-2 may play an import role in vasculogenic mimicry (VM) through regulation of EMT. Moreover, regulators of EMT and VM are known to be important targets for microRNAs (miRNAs). To better understand how these critical pathways are induced by coexpression of Twist-1 and Bcl-2, we performed a comprehensive comparative bioinformatics analysis using microarrays on HCCs that overexpressed Twist-1 and Bcl-2. Eleven miRNAs associated with coexpression of Twist-1 and Bcl-2 were selected from the comprehensive analysis of miRNA microarray and ChIP-seq analysis. Changes in miRNAs were associated with significant differences in the expression of genes involved in signal transduction pathways related to processes including tumor invasion, metastasis, angiogenesis, and tumor cell shape. We confirmed the role of Twist-1 and Bcl-2 coexpression in HCC cells using wound healing assays, invasion assays, and 3D Matrigel assays. Furthermore, the role of miR-27a as a crucial regulator of EMT and VM was confirmed in HCC cells by RT-PCR and western blot analysis. These findings provide evidence that Bcl-2 enhances the role of Twist-1 in VM and EMT through miRNAs.

The activation of HO-1/Nrf-2 contributes to the protective effects of diallyl disulfide (DADS) against ethanol-induced oxidative stress.

BACKGROUND: Diallyl disulfide (DADS) is a garlic-derived organosulfur compound. The current study is designed to evaluate the protective effects of DADS against ethanol-induced oxidative stress, and to explore the underlying mechanisms by examining the HO-1/Nrf-2 pathway. METHODS: We investigated whether or not DADS could activate the HO-1 in normal human liver cell LO2, and then evaluated the protective effects of DADS against ethanol-induced damage in LO2 cells and in acute ethanol-intoxicated mice. The biochemical parameters were measured using commercial kits. HO-1 mRNA level was determined by RT-PCR. Histopathology and immunofluorescence assay were performed with routine methods. Protein levels were measured by western blot. RESULTS: DADS significantly increased the mRNA and protein levels of HO-1, stimulated the nuclear translocation of Nrf-2 and increased the phosphorylation of MAPK in LO2 cells. The nuclear translocation of Nrf-2 was abrogated by MAPK inhibitors. DADS significantly suppressed ethanol-induced elevation of lactate dehydrogenase (LDH) and aspartate transaminase (AST) activities, decrease of glutathione (GSH) level, increase of malondialdehyde (MDA) levels, and apoptosis of LO2 cells, which were all blocked by ZnPPIX. In mice, DADS effectively suppressed acute ethanol-induced elevation of aminotransferase activities, and improved liver histopathological changes, which might be associated with HO-1 activation. CONCLUSION: These results demonstrate that DADS could induce the activation of HO-1/Nrf-2 pathway, which may contribute to the protective effects of DADS against ethanol-induced liver injury. GENERAL SIGNIFICANCE: DADS may be beneficial for the prevention and treatment of ALD due to significant activation of HO-1/Nrf-2 pathway.

NLRP3 inflammasome activation triggers severe inflammatory liver injury in N, N-dimethylformamide-exposed mice.

N,N-dimethylformamide (DMF) is a widely utilized chemical solvent with various industrial applications. Previous studies have indicated that the liver is the most susceptible target to DMF exposure, whereas the underlying mechanisms remain to be elucidated. This study aimed to investigate the role of NLRP3 inflammasome in DMF-induced liver injury in mice by using two NLRP3 inflammasome inhibitors, Nlrp3-/- mice, Nfe2l2-/- mice, and a macrophage-depleting agent. RNA sequencing revealed that endoplasmic reticulum (ER) stress and NLRP3 inflammasome-associated pathways were activated in the mouse liver after acute DMF exposure, which was validated by Western blotting. Interestingly, DMF-induced liver injury was effectively suppressed by two inflammasome inhibitors, MCC950 and Dapansutrile. In addition, knockout of Nlrp3 markedly attenuated DMF-induced liver injury without affecting the metabolism of DMF. Furthermore, silencing Nfe2l2 aggravated the liver injury and the NLRP3 inflammasome activation in mouse liver. Finally, the depletion of hepatic macrophages by clodronate liposomes significantly reduced the liver damage caused by DMF. These results suggest that NLRP3 inflammasome activation is the upstream molecular event in the development of acute liver injury induced by DMF.

The roles of garlic on the lipid parameters: a systematic review of the literature.

Garlic has long been the focus of experimental and clinical attentions due to its promising lipid-lowering effects. Numerous animal studies as well as in vitro ones have demonstrated the hypolipidemic effects of garlic, while clinical trials are highly inconsistent. Based on some double-blind, randomized, placebo-controlled clinical trials which denied the hypolipidemic effects of garlic, some meta-analysis concluded that garlic did not possess beneficial effects for hyperlipidemia. However, we should not ignore the abundant supporting data in the literature. It should be noted that the doses of garlic used in clinical trials were usually far lower than those used in animal studies, which might cover its potential effects. The type of the garlic products may be another important factor responsible for the conflicting outcomes, as different garlic products are composed of different organosulfur compounds. In addition, the biological availability of garlic products is of importance, which was omitted in many studies. Moreover, some studies indicated that different people might have a different response to garlic, and thus garlic may be more beneficial for some specific groups. Collectively, it may be inappropriate to draw a conclusion that garlic does not benefit for hyperlipidemia. Future studies with larger samples are needed to further clarify the effects of garlic used at higher but non-toxic doses on specific groups.

[Effect of docosahexaenoic acid and nervonic acid on the damage of learning and memory abilities in rats induced by 1-bromopropane].

OBJECTIVE: To investigate the protective effects of docosahexaenoic acid (DHA) and nervonic acid (NA) on the learning and memory abilities in rats exposed to 1-bromopropane (1-BP) and their action mechanisms. METHODS: Forty male Wistar rats (specific pathogen-free) were randomly divided into 4 groups (n = 10 for each), i.e., solvent control group, 1-BP (800 mg/kg) group, NA (150 mg/kg) + 1-BP (800 mg/kg) group, and DHA (500 mg/kg) + 1-BP (800 mg/kg) group. The rats were given respective test substances by gavage for 7 d. The Morris water maze (MWM) test was performed from days 8 to 12 to evaluate the rats' learning and memory abilities. After MWM test, rats were sacrificed in the next day, and cerebral cortex was quickly dissected and homogenized in an ice bath. The supernatant of the obtained homogenate was collected to measure the content of glutathione (GSH) and malondialdehyde (MDA) and the activities of glutathione reductase (GR) and γ-glutamate cysteine ligase (γ-GCL). RESULTS: The MWM spatial navigation test showed that the 1-BP group had significantly longer escape latency and significantly longer total swimming distance compared with the control group (P<0.05), while the DHA+1-BP group had significant decreases in escape latency and total swimming distance compared with the 1-BP group (P<0.05). The spatial probe test showed that the number of platform crossings was significantly greater in the DHA+1-BP group and NA+1-BP group than in the 1-BP group (P<0.05); compared with the control group, the 1-BP group had a significantly lower ratio of time spent in the zone around the platform to total time (P < 0.05), and the ratio was significantly higher in the DHA+1-BP group than in the 1-BP group (P < 0.05). Compared with the control group, the 1-BP group had a 18.1% decrease in GSH content, and DHA could significantly reverse 1-BP-induced decrease in GSH content (P < 0.05). Compared with the 1-BP group, the DHA+1-BP group and NA+1-BP group had significantly decreased MDA content (P < 0.05), the DHA+1-BP group had significantly increased GR activity (P < 0.05), and the NA+1-BP group had significantly increased γ-GCL activity (P < 0.05). CONCLUSION: The rats exposed to 1-BP have oxidative stress in the brain and impaired cognitive function. DHA and NA can reduce 1-BP-induced cognitive function impairment in rats, possibly by increasing the activities of GR and γ-GCL and the content of GSH in the brain.

The potential health risks of short-chain chlorinated paraffin: A mini-review from a toxicological perspective.

Short-chain chlorinated paraffins (SCCPs) are ubiquitously distributed in various environmental matrics due to their wide production and consumption globally in the past and ongoing production and use in some developing countries. SCCPs have been detected in various human samples including serum, milk, placenta, nail, and hair, and internal SCCP levels were found to be positively correlated with biomarkers of some diseases. While the environmental occurrence has been reported in a lot of studies, the toxicity and underlying molecular mechanisms of SCCPs remain largely unknown. The current tolerable daily intakes (TDIs) recommended by the world health organization/international programme on chemical safety (WHO/IPCS, 100 µg/kg bw/d) and the UK Committee on Toxicity (COT, 30 µg/kg bw/d) were obtained based on a no observed adverse effect level (NOAEL) of SCCP from the repeated-dose study (90 d exposure) in rodents performed nearly 40 years ago. Importantly, the health risks assessment of SCCPs in a variety of studies has shown that the estimated daily intakes (EDIs) may approach and even over the established TDI by UK COT. Furthermore, recent studies revealed that lower doses of SCCPs could also result in damage to multiple organs including the liver, kidney, and thyroid. Long-term effects of SCCPs at environmental-related doses are warranted.

Promotion of tumor cell metastasis and vasculogenic mimicry by way of transcription coactivation by Bcl-2 and Twist1: a study of hepatocellular carcinoma.

UNLABELLED: The antiapoptotic protein Bcl-2 plays multiple roles in apoptosis, immunity, and autophagy. Its expression in tumors correlates with tumor grade and malignancy. The recapitulation of the normal developmental process of epithelial-mesenchymal transition (EMT) contributes to tumor cell plasticity. This process is also a characteristic of metastatic cells and vasculogenic mimicry. In the present study we report functional and structural interactions between Bcl-2 and the EMT-regulating transcription factor Twist1 and the relationship with metastasis and vascular mimicry. Bcl-2 and Twist1 are coexpressed under hypoxia conditions. The Bcl-2 can bind to Twist1 in vivo and in vitro. This interaction involves basic helix-loop-helix DNA binding domain within Twist1 and through two separate domains within Bcl-2 protein. Formation of the Bcl-2/Twist1 complex facilitates the nuclear transport of Twist1 and leads to transcriptional activation of wide ranges of genes that can increase the tumor cell plasticity, metastasis, and vasculogenic mimicry. Finally, nuclear expression of Bcl-2 and Twist1 is correlated with poor survival of these patients in a cohort of 97 cases of human hepatocellular carcinoma. CONCLUSION: The results describe a novel function of Bcl-2 in EMT induction, provide insight into tumor progression, and implicate the Bcl-2/Twist1 complex as a potential target for developing chemotherapeutics.

A meta-analysis of randomized, double-blind, placebo-controlled trials for the effects of garlic on serum lipid profiles.

BACKGROUND: Inconsistent results were obtained for the lipid-regulating effects of garlic in clinical trials. With increasing interest in complementary medicine for hyperlipoidemia, it is important to explore the real effects of garlic. This meta- analysis was performed to investigate the influence of garlic on serum lipid parameters. RESULTS: A total of 26 studies were included into meta-analysis. Overall, garlic was superior to placebo in reducing serum total cholesterol (TC) and triglyceride (TG) levels. Compared with the placebo groups, serum TC and TG levels in the garlic group were reduced by 0.28 (95% CI, -0.45, -0.11) mmol L⁻¹ (P = 0.001) and 0.13 (95% CI, -0.20, -0.06) mmol L⁻¹ (P < 0.001), respectively. The effects of garlic were more striking in subjects with long-term intervention and higher baseline TC levels. Garlic powder and aged garlic extract were more effective in reducing serum TC levels, while garlic oil was more effective in lowering serum TG levels. In contrast, garlic did not influence other lipid parameters, including low-density lipoprotein cholesterol, high-density lipoprotein cholesterol (HDL-C), apolipoprotein B, and TC/HDL-C ratio. CONCLUSION: Garlic could reduce serum TC and TG levels, and garlic therapy should benefit patients with risk of cardiovascular diseases.

[Development of peripheral neuropathy rat model induced by 1-bromopropane].

OBJECTIVE: To observe the peripheral neurotoxicity of 1-bromopropane (1-BP) by developing an animal model of peripheral neuropathy through oral administration of 1-BP. METHODS: Forty male Wistar rats were randomly and equally divided into low-dose group (200 mg/kg), medium-dose group (400 mg/kg), high-dose group (800 mg/kg), and control group. The rats in the low-dose, medium-dose, and high-dose groups were orally given 1-BP (dissolved in corn oil), while the rats in the control group were orally given an equal volume of corn oil. The oral administration (0.2 ml/100 g BW) was performed once per day, 5 days per week, for 16 consecutive weeks. Neurobehavioral indices including gait score, hindlimb grip strength, and hindlimb landing foot splay were recorded periodically. Hematological and biochemical parameters were also measured during and after 1-BP exposure. RESULTS: The gait scores were significantly higher in the high-dose group (after 8 ∼ 16 weeks of 1-BP exposure), medium-dose group (after 14 ∼ 16 weeks of 1-BP exposure), and low-dose group (after 15 ∼ 16 weeks of 1-BP exposure) than in the control group (P < 0.05, P < 0.01). Compared with the control group, the high-dose group showed significantly decreased hindlimb grip strength after 9, 12, and 14 weeks of 1-BP exposure (P < 0.05, P < 0.01), with the hindlimbs paralyzed after 16 weeks of 1-BP exposure. After 16 weeks of 1-BP exposure, the hindlimb grip strengths of rats in the medium-dose and low-dose groups were decreased to 72.6% and 91.2% of the control value (P < 0.01, P < 0.05). Compared with the control group, the high-dose group showed significantly increased hindlimb landing foot splay after 12, 14, and 16 weeks of 1-BP exposure, and the medium-dose group showed significantly increased hindlimb landing foot splay after 14 and 16 weeks of 1-BP exposure (P < 0.05, P < 0.01). The high-dose and medium-dose groups showed significantly higher serum alanine aminotransferase (ALT) activity than the control group after 8 weeks of 1-BP exposure, and so did the low-dose group after 16 weeks of 1-BP exposure (P < 0.01). CONCLUSION: The nervous system is sensitive to the toxic effect of 1-BP, and 1-BP exposure can induce peripheral neuropathy in rats.

[Spatial Characterization of Stable Isotope Composition of Organic Carbon from Farmland Soils in Chongqing].

Soil was sampled from 182 profiles in typical farmlands of Chongqing and analyzed for the stable carbon isotope composition of organic matter (δ13CSOC). The results showed that the values of δ13CSOC for each soil profile were gradually increasing with increasing soil depth, and the mean values were (-23.63±1.53)‰, (-22.43±1.59)‰, and (-21.42±1.90)‰ for surface, middle, and bottom layers, respectively. The δ13CSOC values in the northeastern region of Chongqing tended to be more negative, whereas those in central Chongqing were less negative. Paddy fields showed the most negative values of δ13CSOC, followed by rice-upland rotating fields and upland fields, with the average being (-25.32±0.93)‰, (-23.17±1.37)‰, and (-24.75±1.28)‰ for the surface layers, respectively. For different soil types, the δ13C values in the surface layers were in the order of paddy soil

[Effects of Biodegradable Film Raw Material Particles on Soil Properties, Wheat Growth, and Nutrient Absorption and Transportation].

Biodegradable plastic film is one of the effective ways to solve the problem of white pollution in agriculture. However, its impacts on soil-plant systems are not well documented. In order to provide a basis for the safety evaluation of large-scale application of biodegradable plastic film, pot experiments were conducted to investigate the effects of the types(H, S, and X) and doses(2.5, 10, and 40 g·kg-1) of biodegradable film raw material particles on the soil physiochemical properties, biological properties, growth, and nutrient absorption by wheat (Triticum aestivum L.). The results showed that three types of biodegradable film raw material particles significantly increased soil pH but had no significant effect on soil organic matter content; medium-high doses of H and low-medium doses of S plastic particles had a positive effect on soil nitrification and soil nitrogen availability, whereas X film particles had an inhibitory effect. H film particles increased soil available phosphorus content, and S and X had no significant effect. X film particles increased the content of soil available potassium, but S and H had no significant effect. The effects of three types of biodegradable raw material particles on soil enzyme activities varied with the types and doses of plastic film and enzyme types. With the increase in the doses of plastic film particles, the activities of three kinds of soil enzymes showed a downward trend. Except for the low and medium doses of the S treatment, the other treatments inhibited the growth of wheat, in which X film particles had the greatest inhibitory effect on the biomass of wheat roots, stems, leaves, and grain; with the increase in the doses of film particles, the inhibition effect of wheat biomass was more obvious. For wheat nutrients, the absorption of nitrogen was promoted at low doses and inhibited at high doses, and the three types of film particles inhibited the absorption of phosphorus and potassium. There were significant differences in the distribution ratio of nitrogen and phosphorus between the stems, leaves, and grains of wheat by all the film particles; however, there was no significant difference in the distribution ratio of potassium between those treatments. Correlation analysis showed that wheat biomass was the main factor affecting wheat nutrient accumulation.

Promotion of hepatocellular carcinoma metastasis through matrix metalloproteinase activation by epithelial-mesenchymal transition regulator Twist1.

E-cadherin loss is a key biological mechanism in tumour invasion. As a main regulator of epithelial-mesenchymal transition (EMT) mechanism-mediated invasion and metastasis, Twist1 plays an important role through its regulation of E-cadherin expression. However, whether or not Twist2 has the same function in tumour metastasis remains unclear. The purpose of this study is to investigate the expressions and different roles of Twist1 and Twist2 in human hepatocellular carcinoma (HCC). The expressions of Twist1 and Twist2 in HCC tissue were evaluated by immunohistochemical staining. The role of Twist1 and Twist2 in invasiveness was also evaluated in vitro by using HCC cell lines. Twist1 nuclear overexpression is found to be correlated with HCC metastasis, and its expression is negatively correlated with E-cadherin expression in human tissue. Twist2, a Twist1 homology protein, only expresses in the cytoplasm and shows no significant correlation with HCC metastasis. By ectopic transfection of Twist1 and Twist2 into the HCC cells, HepG2 and PLC, Twist1 is able to down-regulate E-cadherin expression and promote matrix metalloproteinase (MMP) activation, specifically in MMP2 and MMP9. In functional assays, Twist1 is found to promote invasion in HepG2 and PLC cells, but the invasion ability of the groups is not affected Twist2. Our findings indicate that Twist1 induces HCC invasion via increased activity in MMPs, leading to poor clinical prognoses. The results of this study also demonstrate a novel cogitation in Twist2, which has no effect on HCC invasion and metastasis. Twist1 may contribute to HCC invasion and metastasis and may be used as a novel therapeutic target for the inhibition of HCC metastasis.

Expression and functional significance of Twist1 in hepatocellular carcinoma: its role in vasculogenic mimicry.

UNLABELLED: The up-regulation and nuclear relocation of epithelial-mesenchymal transition (EMT) regulator Twist1 have been implicated in the tumor invasion and metastasis of human hepatocellular carcinoma (HCC). The term vasculogenic mimicry (VM) refers to the unique capability of aggressive tumor cells to mimic the pattern of embryonic vasculogenic networks. However, the relationship between Twist1 and VM formation is not clear. In this study, we explored HCC as a VM and EMT model in order to investigate the role of Twist1 in VM formation. We first examined the expression of Twist1 in human HCC samples and cell lines and found that Twist1 was frequently overexpressed in the nuclear relocation occurring in VM-positive HCCs (13/18 [72%]). Twist1 nuclear expression was likewise significantly associated with VM formation. Clinicopathological analysis revealed that both VM and Twist1 nuclear expressions present shorter survival durations than those without expression. We consistently demonstrated that an overexpression of Twist1 significantly enhanced cell motility, invasiveness, and VM formation in an HepG2 cell. Conversely, a knockdown of Twist1 by the short hairpin RNA approach remarkably reduced Bel7402 cell migration, invasion, and VM formation. Using chromatin immunoprecipitation, we also showed that Twist1 binds to the vascular endothelial (VE)-cadherin promoter and enhances its activity in a transactivation assay. CONCLUSION: The results of this study indicate that Twist1 induces HCC cell plasticity in VM cells more through the suppression of E-cadherin expression and the induction of VE-cadherin up-regulation than through the VM pattern in vivo and in a three-dimensional in vitro system. Our findings also demonstrate a novel cogitation in cancer stem-like cell differentiation and that related molecular pathways may be used as novel therapeutic targets for the inhibition of HCC angiogenesis and metastasis.

[The effects of tea polyphenols on the injury of fibrinolytic functions induced by high-methionine dietary in rats].

OBJECTIVE: To study the protective impact of tea polyphenols (TP) on the injury of fibrinolytic functions induced by high-methionine dietary in rats. METHODS: 50 male Wistar rats were divided by stratified based on body weight into 5 groups with 10 in each group: namely control group, model group, low-dose TP group, medium-dose TP group and high-dose TP group. The rats in model group and TP groups were fed with 3% methionine dietary, control group rats with routine diet. In addition, rats in low-dose, medium-dose and high-dose TP groups were treated with TP at 50, 100 and 200 mg/kg dosage respectively by gavages every day, control group and model group rats were given with same amount distilled water. The animals were sacrificed after 8 weeks. The levels of tissue-type plasminogen activator (t-PA) and type-1 plasminogen activator inhibitor (PAI-1) in plasma were determined by ELISA assays, mRNA levels of t-PA and PAI-1 in aortic arch were detected by RT-PCR, t-PA and PAI-1 expression in aortic arch were detected by immunohistochemistry strept-avidin-biotin complex (SABC). RESULTS: After experiment, the t-PA expression of aortic arch in control group, model group, low-dose TP group, medium-dose TP group and high-dose TP group were 133.03 ± 10.14, 95.46 ± 11.08, 111.97 ± 11.91, 130.23 ± 10.80, 139.39 ± 9.41 (F = 14.15, P < 0.01), respectively, and the PAI-1 expression were 90.91 ± 8.67, 166.76 ± 12.18, 139.63 ± 12.71, 134.66 ± 13.19, 109.49 ± 10.82 (F = 31.44, P < 0.01). The t-PA concentration of plasma were (10.69 ± 1.26), (6.13 ± 0.92), (8.56 ± 1.19), (9.69 ± 0.92), (11.97 ± 1.08) ng/ml, respectively (F = 41.98, P < 0.01), and the PAI-1 concentration of plasma were (6.31 ± 0.81), (16.98 ± 1.27), (11.39 ± 0.82), (8.46 ± 0.67), (8.08 ± 0.91) ng/ml, respectively (F = 207.74, P < 0.01). The mRNA levels of t-PA in aortic arch were 1.12 ± 0.02, 0.75 ± 0.14, 1.01 ± 0.09, 0.95 ± 0.08, 1.05 ± 0.13 (F = 5.77, P < 0.05), and the mRNA levels of PAI-1 in aortic arch were 1.25 ± 0.11, 1.74 ± 0.06, 1.23 ± 0.05, 1.09 ± 0.14, 1.23 ± 0.04 (F = 23.56, P < 0.01). CONCLUSION: The results indicate that TP seems to have regulatory function on transcription and protein levels of t-PA and PAI-1, in addition to maintaining the balance between PAI-1 and t-PA and healing the injury of fibrinolytic functions in rats induced by high-methionine dietary.

[Effects of 1-bromopropane exposure on cognitive function in rats].

OBJECTIVE: To study the effects of 1-bromopropane (1-BP) on the functions of learning-memory and the central cholinergic system in rats. METHODS: Forty male Wistar rats were randomly divided into four groups: low 1-BP group (200 mg/kg), middle 1-BP group (400 mg/kg), high 1-BP group (800 mg/kg) and control group, and the exposure time was 7 days. The Morris water maze (MWM) test was applied to evaluate the learning-memory function in rats. After the MWM test, the rats were sacrificed, the cerebral cortex and hippocampus were quickly dissected and homogenized in ice bath. The activity of acetylcholine esterase (AChE) and choline acetyltransferase (ChAT) in supernatant of homogenate were detected. RESULTS: The latency and swim path-length of rats in middle and high 1-BP groups prolonged significantly in place navigation test and the efficiency of searching strategy obviously decreased, as compared with control group (P < 0.05 or P < 0.01). In spatial probe test, the number of crossing platform in three 1-BP groups decreased significantly, as compared with control group (P < 0.05 or P < 0.01). The cortical AChE activity of rats in middle and high 1-BP groups was significantly higher than that of control and low 1-BP group (P < 0.05 or P < 0.01). The AChE activity in rat hippocampus of high 1-BP group obviously increased, as compared with control group as compared with control group (P < 0.05). There was no significant difference of cortical ChAT activity between three 1-BP groups and control group (P > 0.05). In the hippocampus, there was no difference of ChAT activity among the groups (P > 0.05). CONCLUSION: 1-BP exposure could significantly influence the learning-memory function in rats due to the increase of AChE activity.

[Effects of Water Management and Silicon Application on Iron Plaque Formation and Uptake of Arsenic and Cadmium by Rice].

To explore the effects of water management and silicon application on the bioavailability of soil arsenic (As) and cadmium (Cd), and the accumulation of As and Cd in rice, pot experiments were carried out using As/Cd co-contaminated paddy soil from a field in Kaiyang County, Guizhou Province. The experimental treatments had the following five water application modes with and without silicon addition:flooding during the entire growth period (T1); flooding for three weeks (0-105 d) after transplanting with wet irrigation (moisture content 50%-60%) during other growth periods (T2); flooding for three weeks before heading (0-65d), three weeks after heading (84-105d), and wet irrigation during other growth periods (T3); flooding from heading to three weeks after heading (84-105d) and wet irrigation during the other growth periods (T4); and wet irrigation during the entire growth period (T5). The results showed that compared with flooding and wet irrigation, flooding combined with wet irrigation was more conducive to the formation of iron plaque (DCB-Fe) on the surfaces of roots. As and Cd content increased with an increasing of content of DCB-Fe. Silicon application increased soil pH and the content of DCB-As but decreased available As and Cd in the soil and, with the exception of the flooding treatment, the DCB-Fe/Cd content. The shorter the flooding time, the higher the accumulation of Cd and the lower the accumulation of As in each part of the rice. Silicon application increased the biomass of rice but decrease the Cd content of roots, stems, leaves, and grain by 4.23%-31.06%, 11.41%-52.90%, 1.74%-35.73%, and 19.25%-39.76%, respectively. Silicon application also decreased the As content of roots, stems, leaves, and grain by 1.47%-52.60%, 6.12%-63.02%, 2.97%-28.41%, and 16.33%-61.23%, respectively. Among the five modes of water application tested, silicon application combined with the T3 mode achieved the highest rice biomass and lowest rice As and Cd contents. Therefore, it is suggested that selecting water management and silicon application regimes according to the level of As/Cd pollution can effectively decrease the bioavailability of As/Cd in the soil, thereby reducing the accumulation of As/Cd in rice.

[Effects of Different Organic Materials on Absorption and Translocation of Arsenic and Cadmium in Rice].

A pot experiment was carried out to study the impacts of five organic materials (rape straw, broad bean stalk, peat, pig manure compost, and biochar) on the availability of arsenic (As) and cadmium (Cd) in soil, the amount of iron plaque on the root surface, as well as the uptake and translocation of As and Cd in rice grown in an As/Cd co-contaminated yellow paddy soil. The results indicated that the application of organic materials significantly increased the contents of the soil organic matter and the yield of rice. The application of broad bean stalk, peat, pig manure compost, and biochar remarkably increased the soil pH, while the application of rape straw exerted no significant influence. The addition of organic matter reduced the available Cd content by 34.77%-82.69%. However, the effects of organic materials on the availability of As varied with the organic materials. The soil-available As content was significantly increased by the application of pig manure compost and biochar, while it was significantly decreased by adding rape straw and peat. The application of organic materials increased As and Cd contents in the Fe plaques on rice root surface by 28.49%-94.86% and 17.73%-151.03%, respectively. It also reduced the As and Cd contents in brown rice by 27.04%-82.51% and 15.87%-79.45%, respectively. The largest decrease was observed in the biochar treatment. The application of organic materials also remarkably reduced the translocation efficiency of Cd from the root-stem-leaf-grain and that of As from the stem to grain. The correlation analysis revealed that the soil pH, available Cd, and Cd content in the Fe plaques are the major factors influencing the accumulation of Cd in the rice grain. Furthermore, the soil pH, soil organic matter, and As content in the Fe plaques are the major factors influencing the accumulation of As in the rice grain. Therefore, it has been concluded that organic materials could influence the uptake and translocation of As and Cd in rice through changing the soil pH, organic matter content, and As and Cd contents in the Fe plaques.

[Analysis of mitochondrial DNA D-loop region mutation and its significance in human oncocytoma].

OBJECTIVE: To investigate the mutation in mitochondrial DNA displacement-loop (mtDNA D-loop) region in oncocytoma and its relationship with tumorigenesis and tumor development. METHODS: The mtDNA D-Loop region of 20 thyroid or renal oncocytomas and the adjacent normal tissues were amplified by PCR, and then sequenced. Five human fetal renal tissues were collected as matched controls. RESULTS: Among the 20 oncocytomas, 21 mutations which focused on hypervariable region I (HVI) were found in 7 tumor tissues and 1 normal tissue with the mutation rates of 35% and 5%, respectively. At the same time, 191 polymorphisms were found in the 20 cases. CONCLUSION: mtDNA D-loop region, especially HV I, is the mutational hotspot of oncocytomas, which may be closely related with mtDNA duplicating rate and the function of mitochondria.