Apoptotic Effects of Hypocrellin A on HeLa Cells

Hypocrellin A（ HA）, a photosensitive perylenequinone compound of Hypocrella bambusae, inhibited the proliferation of several tumor cell lines. Human cervical cancer cells, HeLa ceils, were used as a model to elucidate the molecular mechanisms of HA-induced tumor cell death. The results show that HA can induce the oligonucleosomal fragmentation of DNA in HeLa cells and also can increase the expression of apoptosis inducer Bax mRNA and that it decreases the expression of apoptosis suppressor, Bcl-2 mRNA, in mitochondria. It can be concluded from the data that HA-induced apoptosis is related to the balance between Bcl-2 and Bax gene expressions.

Chemical Chaperones Increasing Expression Level of Soluble Single-chain Fv Antibody(scFv2F3)

Glutathione peroxidase（GPX） is a sclenoenzyme that protects the biomembrane and other cellular components against oxidative damage. The selenium-containing single chain Fv fragment of monoelonal antibody 2173 （Se scFv2F3） is a kind of GPX mimic and it has a wide clinical applications because of its high activity and low antigenicity. Se-seFv2F3 is generated by the chemical modification of the single chain Fv fragment of monoelonal antibody 2F3 （ scFv2F3 ）, which can be expressed in E. coll. In this article, the effect of chemical chaperones, such as glycerol, glucose, and β-cyclodextrin added to the culture medium, on the expression of soluble scFv2F3 was investigated. The expression level was evaluated by the determination of soluble scFv2F3 contents in the whole cell lysates. The results suggest that both glycerol and β-cyclodextrin greatly increase the expression level of soluble scFv2F3, and β-cyclodextrin is found to be more effective compared with glycerol. Glucose has a slight effect on the expression level of soluble scFv2F3. This is the first example, wherein β-cyclodextrin has been used as a chemical chaperone during the cell culture to improve the expression level of recombinant proteins. In addition, chemical chaperones are found to decrease the toxic effect of IPTG on cells.

Effect of Polysaccharides from Fruit Body of Ganoderma tsugae on Bidirectional Regulation of Proinflammatory Cytokine Production in THP-1 Cells

The polysaccharides,FIII-2,FIII-2-a,FIII-2-b and FIII-2-c were purified from the fruit body of Ganoderma tsugae（G.tsugae） to study the effects of the four polysaccharides on the productions of human proinflammatory cytokines,interleukin-1α（IL-1α） and tumor necrosis factor α（TNFα）.The results show that the polysaccharides up-regulated the production of IL-1α and TNFα in THP-1 cells under non-stimulatory condition or under stimulation of both lower concentration lipopolysaccharide（LPS） and phorbol myristate acetate（PMA）,and they down-regulated the secretion of IL-1α and TNFα under stronger stimulatory conditions.The four polysaccharides promoted the expression of TNFα mRNA.Production of TNFα protein reflects the quantity of TNFα mRNA.These polysaccharides purified from Ganoderma tsugae bodies exhibit bidirectional regulation effects on cytokine production under different cell stimulatory conditions.

Antitumor Active Protein-containing Glycans from the Body of Ganoderma tsugae

To explore the effects of traditional herbal medicine Ganoderrna tsugae（G, tsugae） on immunomodulatory and antitumor activities, the crude polysaccharides of G. tsugae were purified by filtration, diethylaminoethyl（DEAE） sepharose-fast flow chromatography and sephadex G-100 size-exclusion chromatography. Two main fractions, pro- tein-containing glyeans CSSLP-1 and CSSLP-2, were obtained via the gradient elution. The protein content, molecu- lar weight, and monosaccharide composition of the two fractions were analyzed. Furthermore, the influence of the protein-containing glycans from G. tsugae on the activation of human acute monocytic leukemia cell line（THP-l） and their antitumor activities to the human hepatocellular liver carcinoma cell（HepG-2） in vitro were evaluated. The re- sults indicate that CSSLP-1 and CSSLP-2 could increase the pinocytie activity of THP-1 cells and induce THP-1 cells to produce the eytokines of TNFa and IL-2, significantly. CSSLP-1 and CSSLP-2 also played an inhibiting effect on the cancer cell（HepG-2）. Moreover, the anti-proliferation activity of CSSLP-1 and CSSLP-2 increased with the par- ticipation of TNFa and IL-2 or other antitumor factors induced from THP-1 cells by G. tsugae protein-containing glycan fractions.