Tellimagrandin I is a hydrolysable tannin compound widely present in plants. In this study, the effect of tellimagrandin I on chemically induced erythroid and megakaryocytic differentiation was investigated using K562 cells as differentiation model. It was found that tellimagrandin I not only inhibited the hemoglobin synthesis in butyric acid (BA)-and hemin-induced K562 cells with IC50 of 3 and 40μM, respectively, but also inhibited other erythroid differentiation marker including acetylcholinesterase (AChE) and glycophorin A (GPA) in BA-induced K562 cells. Tellimagrandin I also inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced expression of CD61 protein, a megakaryocytic marker. RT-PCR analysis showed that tellimagrandin I decreased the expression of erythroid genes (γ-globin and porphobilinogen deaminase (PBGD)) and related transcription factors (GATA-1 and NF-E2) in BA-induced K562 cells, whereas tellimagrandin I induced the overexpresison of GATA-2 transcription factor that played negative regulation on erythroid differentiation. These results indicated that tellimagrandin I had inhibitory effects on erythroid and megakaryocytic differentiation, which suggested that tannins like tellimagrandin I might influence the anti-tumor efficiency of some drugs and the hematopoiesis processes.

To elucidate the mechanism of nacre biomineralization, the mantle of Pinctada fucata (P. fucata) from the South China Sea was used. Using the mantle cDNA library and the ESTs we have cloned through suppression subtractive hybridization (SSH), ten novel genes including PFMG1 were obtained through nested PCR. Bioinformative results showed that PFMG1 had a high homology (40%) with Onchocerca VolVulus calcium-binding protein CBP-1 and had two EF-hand calcium-binding domains from the 81st to the 93rd amino acid and from the 98th to the 133rd amino acid in the deduced amino acid sequence. The results of multitissue RT-PCR and in situ hybridization demonstrated the high expression of PFMG1 in the mantle of P. fucata and confirmed the SSH method. The results of GST-PFMG1 on CaCO3 crystallization showed significant effects on nucleation and precipitation of CaCO3. PFMG1 was cloned into the pcDNA.3.1/myc-HisA vector and was subsequently transfected into MC3T3-E1 cells. RT-PCR revealed upregulation of the marker genes related to cell growth, differentiation, and mineralization, and BMP-2, osterix, and osteopontin were upregulated as a result. This research work suggests that PFMG1 plays an important role in the nacre biomineralization, and the SSH method can pave the way for the bulk cloning and characterization of new genes involved in biomineralization in P. fucata and may accelerate research on the mechanism of pearl formation.

Aconitine is an effective ingredient in Aconite tuber, an important traditional Chinese medicine. Aconitine is also known to be a highly toxic diterpenoid alkaloid with arrhythmogenic effects. In the present study, we have characterized the properties of arrhythmic cytotoxicity and explored the possible mechanisms of aconitine-induced cardiomyocytes. Results show that aconitine induces significant abnormity in the spontaneous beating rate, amplitude of spontaneous oscillations and the relative intracellular Ca2+ concentration. Also, mRNA transcription levels and protein expressions of SR Ca2+ release channel RyR2 and sarcolemmal NCX were elevated in aconitine-induced cardiomyocytes. However, co-treatment with ruthenium red (RR), a RyR channel inhibitor, could reverse the aconitineinduced abnormity in intracellular Ca2+ signals. These results demonstrate that disruption of intracellular Ca2+ homeostasis in the cardiac excitation–contraction coupling (EC coupling) is a crucial mechanism of arrhythmic cytotoxicity in aconitine-induced cardiomyocytes. Moreover, certain inhibitors appear to play an important role in the detoxification of aconitine-induced Ca2+-dependent arrhythmias.

A novel human gene full-length cDNA sequence-TSARG2 was identified from a human testis cDNA library using the SRG2 gene (GenBank Accession No. AF395083), which was significantly up-regulated in cryptorchidism, as an electronic probe. TSARG2 was 1223 bp in length. The putative protein encoded by this gene was 305 amino acids with a theoretical molecular weight of 34,751 and isoelectric point of 9.85. The sequence shared no significant homology with any known protein in databases except SRG2. Northern blot analysis revealed that 1.7 kb TSARG2 transcript was detected selectively in human testis. Furthermore, results of in situ hybridization assay confirmed that TSARG2 was expressed in seminiferous tubules, more precisely in spermatogonia and spermatocyte. No mutation was found by PCR-SSCP in 122 cases of azoospermia, severe oligzoospermia, and cryptorchidism. The green fluorescence produced by pEGFP-C1/TSARG2 was detected on the nucleus of COS7 cells after 24 h post-transfection. The pcDNA3.1())/TSARG2 plasmid was constructed and introduced into MCF7 cells by liposome transfection. TSARG2 can accelerate MCF7 cells to traverse the S-phase and enter the G2-phase compared with the control without transfection of TSARG2, which suggested that this gene plays an important role in the development of cryptorchid testis and is a testis-specific apoptosis candidate oncogene.

The effects of two polyphenols, chebulinic acid and tellimagrandin I, on DNA strand breaks mediated by H2O2/Cu(Ⅱ), hydroquinone (HQ)/Cu(Ⅱ) and H2O2/Fe(Ⅱ) in pBR322 plasmid DNA and genomic DNA of cultured MRC-5 human embryo lung fibroblasts were examined. The results demonstrated that chebulinic acid and tellimagrandin I obviously inhibited HQ/Cu(Ⅱ)-and H2O2/Cu(Ⅱ)-mediated pBR322 DNA strand breaks. When MRC-5 cells were treated with HQ/Cu(Ⅱ), the presence of chebulinic acid or tellimagrandin I inhibited HQ/Cu(Ⅱ)-mediated double strand breaks of genomic DNA. The presence of chebulinic acid or tellimagrandin I did not affect the H2O2-and HQ-mediated reduction of Cu(Ⅱ) to Cu(I). Both polyphenols could slightly inhibit H2O2/Fe(Ⅱ)-mediated plasmid DNA strand break at the lower concentration (1-10 lM), but potentiate the DNA strand break at the higher concentration (over 50 lM). These results demonstrated that chebulinic acid and tellimagrandin I possessed antioxidant action in certain conditions and exerted prooxidant action on DNA strand breaks in other conditions.