Inhibitory activity of polysaccharide extracts from three kinds of edible fungi on proliferation of human hepatoma SMMC-7721 cell and mouse implanted S180 tumor

AIM To determine the activities ofpolysaccharide extracts from Flammulina velutipes (Curt. ex Fr. ) Sing (FV), Lentinusedodes (LE) and Agaricus bisporus Sing (AB)on the proliferation of human hepatoma SMMC-7721 cells in vitro and on mouse implanted S-180tumors in vivo.METHODS The polysaccharide extracts were isolated from the fruit bodies of FV, LE and AB by the methods of hot-water extraction, Sevag’sremoval of proteins, ethanol precipitation,trypsin digestion and ethanol fractionalprecipitation. Human hepatoma SMMC-7721 cells were treated with 50 mg/L Polysaccharide extracts, and the mitosis index, mitochondria activity and cell proliferation were detected at different times in both control and experimental groups. The mice with S-180 implanted tumors were injected with the polysaccharide extracts at 24 mg/ kg body weight for 9 d and the tumorweight was measured on the 15th day.RESULTS The mitosis index of hepatoma cells in vitro could be significantly decreased by treatment with the polysaccharide extracts fromthe three kinds of edible fungi (P < 0 .005 ). Thecell numbers and mitochondria activity of SMMC7721 cells treated with polysaccharide extracts were lower than those in control groups (P <0.005). The inhibition rates of polysaccharide extracts against implanted S-180 tumors in mice were 52.8%, 56.6% and 51 .9% respectivelycompared with that in c0ntrol gr0ups.CONCLUSI0N The POIysaccharide extractsfrom the three kinds of edible fungi could inhibitnot only the Cultured malignant cells in vitfO butalso impIanted Sl80 tum0r i0 vivo.

Clinicopathological significance and prognostic value of LRP16 expression in colorectal carcinoma

AIM: To explore the expression of leukemia related protein 16 (LRP16) in colorectal carcinoma, and analyze its correlation with clinicopathologic features and prognosis. METHODS: Immunohistochemistry for LRP16 was performed in 201 cases of colorectal carcinoma and 60 cases of distal normal mucosa. Medical records were reviewed and clinicopathological analysis was performed. RESULTS: LRP16 expression was detected in 117 of 201 cases of the colorectal carcinoma and in 21 cases of 60 distal normal mucosa. The expression of LRP16 in carcinoma was significantly higher than that in normal mucosa (χ 2 = 9.999, P = 0.002). LRP16 protein expression was found in 43.3% (52/120) of carcinoma at stage Ⅰ and Ⅱ , and 80.2% (65/81) of carcinoma at stage Ⅲ and Ⅳ (χ 2 =27.088, P = 0.001). Correlation between LRP16 expression and clinicopathological factors was significant in differentiation (P = 0.010), tumor size (P = 0.001), infiltrative depth (P = 0.000) and distant metastasis (P = 0.027). The difference of median survival time between cancer patients with LRP16 expression (38.0 mo) and those without was statistically significant (105.0 mo, Log rank = 41.455, P = 0.001). The multivariate survival analysis revealed that LRP16 expression was correlated significantly (Cox’s regression: P = 0.001, relative risk = 2.082) with shortened survival in the patients with colorectal cancer. CONCLUSION: The expression of LRP16 is related to the degree of differentiation, invasiveness, metastasis and prognosis of colorectal carcinoma.

Serum deprivation enhances DNA synthesis of human hepatoma SMMC-7721 cells

AIM To determine the relationship between serum deprivation or serum levels and cell proliferation of human hepatoma SMMC 7721 cells. METHODS Human hepatoma SMMC 7721 cells were grown in RPMI 1640 supplemented with 10% fetal calf (FCS) in 5% CO 2 incubator at 37℃ for 24h , and culture media were replaced to serum free or different serum FCS levels (2 5%, 5%, 10%, 20% and 25%). Six h, 12h , 18h and 24h after the culture, the cells were incorporated TdR for 4h . At last TdR incorporation was detected with liquid scintillation counting. RESULTS DNA synthesis of SMMC 7721 cells could be sharply stimulated by short time (6h) serum deprivation (the cpm value of 3H TdR incorporation of cells in serum free was 39 32 fold higher than cells in 25% serum), and the incorporation of 3H TdR was negatively related to the serum levels. Longer time serum starvation ( 12h , 18h and 24h ) also greatly stimulated DNA synthesis, although the cpm value of 3H TdR incroporation was less than that in 6h serum deprivation. Morphology of cells cultured in different serum levels also showed significant difference. CONCLUSIONS Compared with other cell lines such as BEL7404 and Swiss 3T3, human hepatoma SMMC 7721 cells had different response to the serum deprivation. Short time serum deprivation could greatly stimulate DNA synthesis of human hepatoma SMMC 7721 cells. Precautions must be given to the changes of serum levels for the detection of growth factors and drugs using SMMC 7721 cells as a model.

Biodegradation of Leather Waste by Enzymatic Treatment

The treatment of shavings,trimmings and splits of leather waste from tanneries has a potential to generate value-added products. In this study enzymatic treatment of leather waste was performed. This method utilizes alkaline protease produced by Bacillus subtilis in our laboratory by submerged fermentation. Optimum conditions of pH,time duration,temperature and concentration of enzyme were determined for maximum degradation of leather waste. The amount of degradation was measured by the release of amino acid hydroxyproline. Amino acid composition in the hydrolysate obtained by the enzyme hydrolysis was determined. This relative simple biotreatment of leather waste may provide a practical and economical solution.

Mechanisms and significance of lipoprotein(a) in hepatocellular carcinoma

BACKGROUND: The liver plays a key role in the metabolism of plasma apolipoproteins, endogenous lipids and lipoproteins. Hepatocellular carcinoma is one of the most common fatal malignant tumors in China and in other Southeast Asian countries. It has been demonstrated that plasma lipid profiles are changed in liver cancer. DATA SOURCES: A MEDLINE database search was performed to identify relevant articles using the keywords 'hepatocellular carcinoma' and 'lipoprotein(a)'. The search was conducted and research articles were reviewed from 1960 to 2008. RESULTS: Production and homeostasis of lipids, apolipoproteins and lipoproteins depend on the integrity of hepatocellular functions, which ensures normal lipid and lipoprotein metabolism in vivo. When hepatocellular injury or liver cancer occurs these processes can be impaired. It has been suggested that plasma levels of apolipoprotein(a) (apo(a)) and/or lipoprotein (a) (Lp(a)) may be considered as sensitive markers of hepatic impairment. CONCLUSIONS: Plasma levels of apo(a) and Lp(a) display significant correlations with hepatic status. Most studies demonstrated that the plasma levels of apo(a) and Lp(a) can be considered as an additional clinical index of liver function.

CD4(＋)CD25(＋) treg induction by an HSP60-derived peptide SJMHE1 from schistosoma japonicum is TLR2 dependent

Chronic schistosome infection results in the suppression of host immune responses, allowing long-term schistosome survival and restricting pathology.Current theories suggest that Treg play an important role in this regulation.However, the mechanism of Treg induction during schistosome infection is still unknown.The aim of this study was to determine the mechanism behind the induction of CD4(+)CD25(+) T cells by Schistosoma japonicum HSP60(SjHSP60)-derived peptide SJMHE1 as well as to elucidate the cellular and molecular basis for the induction of CD4(+)CD25(+) T cells during S.japonicum infection.Mice immunized with SJMHE1 or spleen and LN cells from naive mice pretreated.with SJMHE1 in vitro all displayed an increase in CD4(+)CD25(+) T-cell populations.Release of IL-10 and TGF-beta by SJMHE1 stimulation may contribute to suppression.Adoptively transferred SJMHE1induced CD4(+)CD25(+) T cells inhibited delayed-type hypersensitivity in BALB / c mice.Additionally, SJMHE1-treated APC were tolerogenic and induced CD4(+) cells to differentiate into suppressive CD4(+)CD25(+) Treg.Furthermore, our data support a role for TLR2 in SJMHE1-mediated CD4(+)CD25(+) Treg induction.These findings provide the basis for a more complete understanding of the S.japonicum-host interactions that contribute to host homeostatic mechanisms, preventing an excessive immune response.

Inhibitory effects of two oligosaccharideson murine melanoma experimental liver metastasis

AIM To observe the effects of a chemically synthesized tetrose and a natural yeast mannan on experimental liver metastasis of mouse melanoma. METHODS After treated with 4mg tetrose (tetrose group) or 4mg mannan (mannan group) for 30 minutes at 37℃, 0 5ml 1×10 6 B16 MBK melanoma cells were injected into the spleen of mice. Fifty five days later, melanoma metastatic nodes on the surface of the liver and in other organs as well as mouse survival time were observed. RESULTS Of the 6 mice in control (B16 cell+PBS) group, 4 died naturally within 55 days, and 2 were killed on the 55th day. All of the 6 mice had metastases in livers, the total number of the melanoma nodes on each liver surface ranged from 2 to 30, with the largest one merging into the whole liver. One mouse had a neoplasm in the remnant site of injection, and 3 had metastases in lungs. In contrast, of the 6 mice in tetrose group, only one died on the 50th day after injection, with 3 metastases in the liver, the largest being 10mm in diameter, the other 5 mice survived until being dissected on the 55th day after injection and had no liver metastasis, but 3 of them had neoplasms in their remnant sites of injection. In mannan group, all of the 6 mice survived and no metastasis was seen except for 2 liver nodes in one mouse with the largest diameter of 1mm. Neither tetrose nor mannan group had metastasis out of the liver, and the weight of liver in the two groups was significantly lower than those in the control group. CONCLUSION Both tetrose and mannan had the effects of preventing melanoma cells from experimental metastasis to and out of the liver, and prolonging the survival time of the mouse.

THE SYNTHESIS AND 2D NMR STUDY OF DELTORPHINS

We synthesized Deltorphin I, Deltorphin II, and their L-Alanine analogs at position 2. The synthesis was carried out with SPPS and Boc strategy. The data, we got from FAB MS, AAA and sequence analysis were in agreement with the calculated values. We studied the 2D NMR for Deltorphin II and [L-Ala(2)]Deltorphin II in DMSO solution. The H-1-H-1 COSY and 2D NOESY spectra were resolved by computer treatment. The conformations of C-fragment tetrapeptides of Deltorphin II and its L-Ala analog might form a 3(10) alpha -helix. The molecular models of these two peptides were established.

Liraglutide improves the inflammatory response in metabolic associated fatty liver disease through stimulator of interferon genes pathway

Objective To investigate the mechanism by which Liraglutide improves the inflammatory response in metabolic associated faty liver disease (MAFLD) by regulating the interferon gene stimulating factor (STING)signaling pathway.Methods 20 male C57BL/6J mice were randomly divided into normal diet group (NC).

Effects of lanthanide ions on hydrolysis of phosphatidylin-ositol in human erythrocyte membranes

The effects of lanthanides on the hydrolysis of phosphatidylinositol in human erythrocyte membranes were studied. 3H-inositoI labeling chromatography and HPLC were used to determine inositol 1, 4, 5-triphosphatc and dia-cylglycerol separately, the hydrolytic products of phosphatidylinositol due to the reaction of lanthanide ions with human erythrocyte membranes. The unhydrolyzed phosphatidylinositol in membranes was also determined. The results indicate that the hydrolysis of phosphatidylinositol can be promoted by lanthanides (La3+, Ce3+, Y3+, Tb3+) and the effects of La3+ and Ce3+ are stronger than those of Y3+ and Tb3+.

Oxalate-induced resistance of muskmelon to WMV-2

The treatment of muskmelon cultivar 'Wangwenxiang', which is sensitive to watermelon mosaic virus-2 (WMV-2), with oxalate increases systemic resistance of the cultivar to WMV-2 significantly. The challenge inoculation experiments indicate that the symptom of ox-alate-treated plant is much slighter than that of control plants and the virus content is only 4%

Apoptosis in suspension culture of tobacco cells induced by heat shock

The induction of apoptosis in suspension culture of tobacco cells by heat shock is reported for the first time. Heat treatment (48℃ for 4 h) of tobacco cells led to the appearance of typical hallmarks of apoptosis. It was demonstrated by DNA laddering analysis that the cells treated with heat shock at 48℃ for 4 h had a serious degradation of nuclear DNA into multi-nu-cleosomal sizes, suggesting that heat shock activated endogenous nuclease which led to DNA cleavage at the linkage sites between the nucleosomes, but ladders were very faint for DNA from 2 and 9 h heat-treated cells. The terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling (TUNEL) detection also showed that most of these treated cells (48℃ for 4 h) displayed positive reactions, indicating a serious DNA 3’-OH cleavage in their nuclei. Moreover, some other cytological changes in apoptotic cells, such as cell shrinkage, chromatin aggregation, nucleus collapse, have also been observed by 4’,

Assaying poly(ADP-ribose) polymerase activity in plants by polarographic method

A new method has been developed to assay poly(ADP-ribose) polymerase (PARP) activity in plant tissues through determining the content of nicotinamide (NIC) produced by enzymatic reaction by linear sweeping polarographic method. The detection limit of NIC was 0.03μmol/L, the calibration graph was linear up to 5μmol/L ( r = 0.999). The recoveries were approximately in the range of 92% to 98% and the relative standard deviations were less

Diatom-based reconstruction of Holocene environmental changes in Angren Lake,Southern Tibet

Based on the diatom analysis the climatic sequence between 11 000 and 6 000 aBP in Angren, southern Tibet is established, i.e. cold period (11 095-9 970 aBP)→temperate period (9 970-9 870 aBP)→Anathermal (9 870-9 770 aBP)→cool-dry period (9 770-8 740 aBP) →super-megathermal (8 740-8 590 aBP)→Megathermal (8 590-5 970 aBP). And the opinion that the reaction of diatom to temperature change is more sensitive than to other ecological factors has been pointed out and certified.

Effects of expressing calmodulin antisense RNA on cell proliferation and transformation in human hepatoma cell lines

AS the main intracellular receptor of Ca2+,calmodulin（CaM）,which is extensively distribut-ed in various types of eukaryotic cells,is a multifunctional protein regulating the cell growth,differentiation and transformation.

Identification of long chain isoprenoid hydrocarbons from pyrolytic product of Dunaliella

In the pyrolytic research on Dunaliella salina cultured under natural conditions, abundant long chain regular isoprenoid hydrocarbons have been first detected and identified from algal pyrolytic product. Specially, the discovery of high abundant 2,6,10,14,18-pentamethyle-icosane indicated that regular iC25 probably originated from specific holophilic algae in hyper-

Effects of low frequency electromagnetic fields on osteoblasts proliferation and cell cycle

After primary mouse osteoblasts and ROS osteo-like cells were exposed to 50 Hz low frequency electromagnetic fields (EMF), the MTT method and flow cytometry have been used to determine cell proliferation, cell cycle and apoptosis. The results show that, compared