Antitumor activity of fucoidan in anaplastic thyroid cancer via apoptosis and anti-angiogenesis.

The present study demonstrated the effect of fucoidan, isolated from Fucus vesiculosus, on cell growth and apoptosis in anaplastic thyroid cancer cells. The cell viability was analyzed using a Cell Counting Kit­8 cell proliferation kit. Diamidino-2-phenylindole and terminal deoxynucleotidyl transferase-mediated dUTP nick­end labeling assays were used to examine the apoptotic effect of fucoidan, which revealed the presence of apoptotic bodies and DNA fragmentation. Fucoidan inhibited the growth of FTC133 and TPC1 ATC cells in a dose­dependent manner. It also induced the apoptosis of FTC133 cells by promoting the expression levels of cleaved poly ADP­ribose polymerase and caspase­3. Significant decreases in the levels expression of hypoxia-inducible factor 1α and vascular endothelial growth factor were observed in the FTC133 cells following treatment of the cells with fucoidan. In addition, inhibition in tube formation and the migration of FTC133 cells were observed in the cells treated with fucoidan, compared with the cells in the control group. Therefore, fucoidan inhibited cell growth, induced apoptosis and suppressed angiogenesis in the thyroid cancer cells.

[Distribution of transferable nitrogen in Poyang Lake sediments and its response to the variation of River-Lake relationship].

Amounts and distributions of total transferable nitrogen and different transferable nitrogen forms were studied in the sediments of Poyang Lake with different regimen, in order to reveal the influence of the water level change caused by the River-Lake relationship change on the potential release risks of nitrogen. The results showed that: (1) the contents of the total nitrogen (TN) were between 389 and 3865 mg x kg(-1), and the spatial distribution showed an overall downward trend in the "Five River", the "Hu Xin" and the northern regions; the contents of the transferable total nitrogen (TTN) were between 319.36 and 904.56 mg x kg(-1) and contributed 52% to the TN, and its spatial distribution trend was the same as that of TN. (2) The content of transferable nitrogen followed the order of SOEF-N approximately = SAEF-N > WAEF-N > IEF-N. (3) The dry period advanced and the low water level continued to decline as a result of the change in River-Lake relationship of the Poyang Lake, leading to the prolonged outcropped time of sediments and the increased area, causing the different contents of transferable nitrogen in the sediments at different elevations. The transferable nitrogen content of the sediments in the whole lake during the wet period was lower than that during the dry period. The content of transferable nitrogen followed the order of 12-13 m elevation sediment > 11-12 m elevation deposition material > 10-11 m elevation sediments. The higher the elevation, the longer the sediments were exposed, and the higher the transferable nitrogen content. (4) With the increase of elevation, all forms of N contents increased. The increase of IEF-N and SOEF-N contents as well as their percentages in total transferable nitrogen was relatively small, while the increase of WAEF-N and SAEF-N contents as well as their percentages in total transferable nitrogen was relatively large. If the river-lake relationship changes further, the elevation in the dry period will further decrease, which will lead to the enlarged area and prolonged exposure of sediments, resulting in increase of TN, transferable nitrogen, IEF-N and SOEF-N contents in the sediments. At the coming wet period of next year, the nitrogen release risk from sediments of Poyang Lake might increase.

High-confidence de novo peptide sequencing using positive charge derivatization and tandem MS spectra merging.

De novo peptide sequencing holds great promise in discovering new protein sequences and modifications but has often been hindered by low success rate of mass spectra interpretation, mainly due to the diversity of fragment ion types and insufficient information for each ion series. Here, we describe a novel methodology that combines highly efficient on-tip charge derivatization and tandem MS spectra merging, which greatly boosts the performance of interpretation. TMPP-Ac-OSu (succinimidyloxycarbonylmethyl tris(2,4,6-trimethoxyphenyl)phosphonium bromide) was used to derivatize peptides at N-termini on tips to reduce mass spectra complexity. Then, a novel approach of spectra merging was adopted to combine the benefits of collision-induced dissociation (CID) and electron transfer dissociation (ETD) fragmentation. We applied this methodology to rat C6 glioma cells and the Cyprinus carpio and searched the resulting peptide sequences against the protein database. Then, we achieved thousands of high-confidence peptide sequences, a level that conventional de novo sequencing methods could not reach. Next, we identified dozens of novel peptide sequences by homology searching of sequences that were fully backbone covered but unmatched during the database search. Furthermore, we randomly chose 34 sequences discovered in rat C6 cells and verified them. Finally, we conclude that this novel methodology that combines on-tip positive charge derivatization and tandem MS spectra merging will greatly facilitate the discovery of novel proteins and the proteome analysis of nonmodel organisms.

[Expression of MCP-1 in the patients of chronic hepatitis B complicated with nonalcoholic fatty liver disease].

AIM: To investigate the expression level of monocyte chemoattractant protein-1 (MCP-1) in the liver of the patients with chronic hepatitis B (CHB) complicated with non-alcoholic fatty liver diseases (NAFLD). METHODS: The study enrolled 21 CHB with NAFLD patients and 46 CHB without NAFLD patients as the controls. Real-time quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR) and immunohistochemistry (IHC) were applied to detect the expression of MCP-1 at the mRNA and protein levels in the liver tissues, respectively. Non-parametric Mann-Whitney test was used to analyze the difference between the CHB patients with and without NAFLD. RESULTS: The mRNA relative expression level of MCP-1 in CHB+NAFLD group was 0.034 (0.024-0.058), higher than that in the control group 0.016 (0.012-0.024). The immunohistochemical score was 8.7±2.5 in CHB+NAFLD group and 6.2±3.5 in the control group. The difference in MCP-1 expression at the both protein and mRNA levels was significant statistically between the two groups (P<0.01). CONCLUSION: MCP-1 expression level in the liver is higher in CHB with NAFLD patients than that in CHB without NAFLD patients.

Expression of the apoptosis-related genes BCL-2 and BAD in human breast carcinoma and their associated relationship with chemosensitivity.

OBJECTIVE: To evaluate the expression of BCL-2 and BAD genes in tissues of breast carcinoma and investigate the relationship between the expression of BCL-2 and BAD in breast cancer cells with chemosensitivity. METHODS: Immunohistochemical technique was used to detect the expression of BCL-2, BAD in 10 normal breast tissues, 10 breast fibroadenoma tissues, 40 youth human breast carcinoma tissues, 40 menopause human breast carcinoma tissues. And to detect the expression of ER, PR in 80 human breast carcinoma tissues. 20 Surgical samples of breast cancer, diagnosed by pathology, were obtained from The First Affiliated Hospital of Chongqing Medical University. The cancer sample cells were cultured separately in the incubator at 37 degrees C, 5% CO2 in vitro. The rate of inhibition of cancer cells in 4 kinds of anticancer drugs-- Epirubicin Adriamycin (EADM),5-Fluorouracil (5-Fu), Navelbine(NVB) and Diaminedichloroplatinum (DDP), were assayed by MTT method. RESULTS: The expression of BCL-2, BAD genes in young human breast carcinoma tissues were lower than that in menopause human breast carcinoma tissues (P < 0.05). There was a negative correlation between the positive expression rate of BCL-2 and histologic grade or the lymph node metastasis (P < 0.05). There was a positive correlation between the expression rates of BCL-2 and of ER, PR (P < 0.05). The expression of BAD had no relationship with the expression of ER, PR, histologic grade and the lymph node metastasis(P = NS). Sensitivity rates of 20 breast cancer cells in 0.1 x PPC within 48 h in vitro were 30% EADM,20% 5-Fu,45% NVB and 25% DDP. Respectively, the rate of inhibition of EADM,5- Fu, NVB and DDP were significantly higher in the BCL-2 negative cancer cells than in the BCL-2 positive cancer cells. A negative correlation was found between expression of BCL-2 and chemosensitivity for all the 4 anticancer drugs. The inhibition rates of EADM and NVB were significantly lower in the BAD negative cancer cells than in the BAD positive cancer cells. A positive correlation was found between expression of BAD and chemosensitivity for Epirubicin. CONCLUSION: The expression of BCL-2 and BAD can be used as prognosis factors of breast cancer. Detection of the BCL-2 protein expression level, particularly, combined with the detection of the expression of BCL-2 and BAD as well as ER and PR were helpful in confirming the prognosis of breast carcinoma. The combined detection of BCL-2 and BAD may be markers for predicting the responses to anticancer drugs.