In vivo and in vitro effect of hepatocarcinoma lymph node metastasis by upregulation of Annexin A7 and relevant mechanisms.

We unveiled the association of Annexin A7 with vascular endothelial growth factor-C (VEGF-C) and the effect of upregulation of Annexin A7 in Hca-F and Hca-P cells on inhibiting hepatocarcinoma (HCC) lymph node metastasis (LNM) in vitro and in vivo. A total of 200 inbred 615 mice were randomly divided into four equal groups inoculated with Hca-F, Hca-P, FAnxa7-upregulated, and PAnxa7-upregulated cells, respectively. The primary tumor, popliteal, inguinal, and iliac lymph nodes were prepared for immunohistochemical (IHC) staining, real-time quantitative polymerase chain reaction (qRT-PCR) analysis, Western blot, and hematoxylin-eosin (H&E) staining. There was over 50 % increase both in the number of FAnxa7-upregulated and PAnxa7-upregulated cells migrated through the filter compared to their controls (FAnxa7-control, Hca-F and PAnxa7-control, Hca-P). However, no significant differences were noted in invasion ability between them (all P > 0.05). Tumor lymph vessels were significantly reduced in FAnxa7-upregulated and PAnxa7-upregulated tumors when compared with Hca-F and Hca-P tumors (all P < 0.05). Blood vessel density did not differ significantly between FAnxa7-upregulated and PAnxa7-upregulated tumors and Hca-F and Hca-P tumors. Enzyme-linked immunosorbent assay (ELISA) for VEGF-C showed that upregulating Annexin A7 decreased VEGF-C secretion in FAnxa7-upregulated and PAnxa7-upregulated cells (P < 0.05). The IHC staining result showed that the level of serum Annexin A7 was found to be statistically higher in all experimental groups than that in the control group (P < 0.05). The present results indicated that alterations in serum Annexin A7 expression may be of prognostic relevance in HCC lymphatic metastasis.

Modified hydrated sodium calcium aluminosilicate-supplemented diet protects porcine oocyte quality from zearalenone toxicity.

Zearalenone (ZEN) is one of the most common mycotoxins produced by fungus in contaminated feed. ZEN has multiple toxicities, including reproductive toxicity of domestic animals, particularly pigs. However, studies on the effects of ZEN on ovary/oocytes have been primarily based on in vitro experiments, and there is still no evidence from porcine in vivo models due to multiple limitations. Moreover, no report has investigated the effect of hydrated sodium calcium aluminosilicate (HSCAS) as a supplement on pig oocyte quality. In the present study, we fed pigs a 1.0 mg/kg ZEN-contaminated diet for 10 days. The results showed that pigs fed ZEN presented reduced oocyte-cumulus cell interactions, an increase in the number of denuded oocytes in ovaries, a decrease in the number of oocytes in each ovary, and an increase in the oocyte death rate. Oocytes from ZEN-exposed pigs exhibited a delayed cell cycle and abnormal cytoskeletal dynamics during meiotic maturation, which could be due to oxidative stress-induced autophagy. Moreover, we also show that supplementing the ZEN-contaminated diet with modified HSCAS effectively protected porcine oocyte quality. Taken together, our study provides in vivo data demonstrating the protective effects of HSCAS against ZEN toxicity in porcine oocytes.

Comparison of the toxic effects of different mycotoxins on porcine and mouse oocyte meiosis.

BACKGROUND: Aflatoxin B1 (AFB1), deoxynivalenol (DON), HT-2, ochratoxin A (OTA), zearalenone (ZEA) are the most common mycotoxins that are found in corn-based animal feed which have multiple toxic effects on animals and humans. Previous studies reported that these mycotoxins impaired mammalian oocyte quality. However, the effective concentrations of mycotoxins to animal oocytes were different. METHODS: In this study we aimed to compare the sensitivity of mouse and porcine oocytes to AFB1, DON, HT-2, OTA, and ZEA for mycotoxin research. We adopted the polar body extrusion rate of mouse and porcine oocyte as the standard for the effects of mycotoxins on oocyte maturation. RESULTS AND DISCUSSION: Our results showed that 10 µM AFB1 and 1 µM DON significantly affected porcine oocyte maturation compared with 50 µM AFB1 and 2 µM DON on mouse oocytes. However, 10 nM HT-2 significantly affected mouse oocyte maturation compared with 50 nM HT-2 on porcine oocytes. Moreover, 5 µM OTA and 10 µM ZEA significantly affected porcine oocyte maturation compared with 300 µM OTA and 50 µM ZEA on mouse oocytes. In summary, our results showed that porcine oocytes were more sensitive to AFB1, DON, OTA, and ZEA than mouse oocytes except HT-2 toxin.

Potent Nematicidal Activity and New Hybrid Metabolite Production by Disruption of a Cytochrome P450 Gene Involved in the Biosynthesis of Morphological Regulatory Arthrosporols in Nematode-Trapping Fungus Arthrobotrys oligospora.

Types of polyketide synthase-terpenoid synthase (PKS-TPS) hybrid metabolites, including arthrosporols with significant morphological regulatory activity, have been elucidated from nematode-trapping fungus Arthrobotrys oligospora. A previous study suggested that the gene cluster AOL_s00215 in A. oligospora was involved in the production of arthrosporols. Here, we report that disruption of one cytochrome P450 monooxygenase gene AOL_s00215g280 in the cluster resulted in significant phenotypic difference and much aerial hyphae. A further bioassay indicated that the mutant showed a dramatic decrease in the conidial formation but developed numerous traps and killed 85% nematodes within 6 h in contact with prey, in sharp contrast to the wild-type strain with no obvious response. Chemical investigation revealed huge accumulation of three new PKS-TPS epoxycyclohexone derivatives with different oxygenated patterns around the epoxycyclohexone moiety and the absence of arthrosporols in the cultural broth of the mutant ΔAOL_s00215g280. These findings suggested that a study on the biosynthetic pathway for morphological regulatory metabolites in nematode-trapping fungus would provide an efficient way to develop new fungal biocontrol agents.

[Expression and clinical significance of Mucin and E-cadherin in colorectal tumors].

BACKGROUND & OBJECTIVE: Mucin (MUC), a glycoprotein with high molecular weight, can lubricate and protect the epithelium. E-cadherin (E-cad) is helpful in keeping the polarity and integrity of the epithelium. The abnormal expression of Mucin and E-cad is involved in the genesis of many tumors. This study was to investigate the expression of MUC1, MUC2, MUC5AC and E-cad in different colorectal tumor tissues, and explore their correlations to clinicopathologic features of colorectal cancer and the correlations of MUC1, MUC2, and MUC5AC expression to E-cad expression. METHODS: The expression of MUC1, MUC2, MUC5AC and E-cad in 150 specimens of normal colorectal mucosa, 150 specimens of colorectal adenoma and 150 specimens of colorectal adenocarcinoma was detected by immunohistochemistry. Patients' survival was analyzed by Kaplan-Meier method. The correlations of MUC1, MUC2, and MUC5AC expression to E-cad expression were analyzed by spearman's rank correlation. RESULTS: The positive rates of MUC1 were 0.07% in normal colorectal mucosa, 12.7% in colorectal adenoma, and 45.3% in colorectal adenocarcinoma. Those of MUC2 were 100%, 90.0% and 52.6%, respectively. Those of MUC5AC were 8.7%, 30.7% and 44.0%, respectively. Those of E-cad were 98.7%, 82.0% and 54.0%, respectively. In colorectal adenocarcinoma, the expression of MUC1 and MUC2 was correlated to tumor differentiation, invasion, lymph node metastasis and Dukes' stage (P<0.05); the expression of MUC5AC was correlated to tumor differentiation and invasion (P<0.01); the expression of E-cad was correlated to tumor differentiation (P<0.01). The 5-year survival rate was significantly higher in MUC1-negative group, MUC2-positive group and E-cadherin-positive group than in their counterparts (P<0.05). In colorectal adenocarcinoma, MUC1 expression was negatively correlated to E-cad expression (r=-0.234, P=0.004), MUC2 and MUC5AC expression were positively correlated to E-cad expression (r=0.170, P=0.038; r=0.198, P=0.015). CONCLUSIONS: In colorectal adenocarcinoma, MUC expression is obviously correlated to E-cad expression. The up-regulation of MUC1 and MUC5AC expression and the down-regulation of MUC2 and E-cad expression may be involved in the genesis of colorectal tumors and reflect the prognosis to a certain extent.