Biosystematics and plant proteomics: role of proteomics in plant phylogenetic analysis.

Since time immemorial, systematics has played significant role in every sphere of life. Biosystematics has evolved from folk taxonomy towards natural classification system and then culminated into homology based classification system. A good systematic approach is practical and predictive of phylogenetics of taxa incorporating different data. The morphological, chemical and molecular (genomics and proteomics) informations are used to explore the exact inter-relationship among the organisms. Proteomics is an essential and inevitable aspect in plant biology which can help in deciphering the functions of the genes that are or will be sequenced. Proteomics has proved to be a good tool in characterisation of individual lines and genetic distances among the genera, species, subspecies, verities and populations describing their phylogenetic interrelationships. Two-dimensional electrophoresis (2-DE) is the major technique being applied for polypeptide characterization of each taxon for exploring phylogenetic or physiological relationships among organs, tissues or organisms. Moreover, proteomics can lead to unraveling the natural phenomena of plants development and their response to changing environment. These proteomic derived informations and their application in phylogenetic studies can be useful in agro-biotechnology development for better yield and safe use of food and medicines.

Involvement of NF-kappaB and c-myc signaling pathways in the apoptosis of HL-60 cells induced by alkaloids of Tripterygium hypoglaucum (levl.) Hutch.

Tripterygium hypoglaucum (levl.) Hutch (Celastraceae) (THH) root is a Chinese medicinal herb commonly used for treating autoimmune diseases. In the present study, alkaloids of THH were prepared and their cytotoxicity against the HL-60 cell was investigated. THH-induced apoptosis was observed using flow cytometry, confocal fluorescence microscope, and DNA laddering and caspase assays. The molecular mechanism involved in the induction of HL-60 cell apoptosis by THH alkaloids was examined using cDNA microarrays containing 3000 human genes derived from a leukocyte cDNA library. Sixteen genes were identified to be differentially expressed in HL-60 cells upon THH treatment. Several genes related to the NF-kappaB signaling pathway and cell apoptosis (such as NFKBIB, PRG1 and B2M) were up-regulated. In addition, c-myc binding protein and apoptosis-related cysteine proteases caspase-3 and caspase-8 were also regulated. The changes in c-Myc RNA expression and c-myc protein level were further confirmed by RT-PCR and Western blot analysis. The results demonstrated that THH alkaloids induced apoptosis of HL-60 cells though c-myc and NF-kappaB signaling pathways.

Arsenic trioxide induces apoptosis of HPV16 DNA-immortalized human cervical epithelial cells and selectively inhibits viral gene expression.

Arsenic trioxide (As2O3), a major ingredient of arsenic compounds in traditional Chinese medicine, exhibits anti-acute promyelocytic leukemic activity. Considering that over 80% of human malignant tumors derive from epithelial cells, we studied the effect of As2O3 on HPV 16 DNA-immortalized human cervical epithelial cells (HCE16/3 cells) in vitro. As2O3 reduced HCE16/3 cell survival, induced apoptosis at a low concentration and selectively inhibited expression of viral early genes. This effect was evidenced by a reduction of cell viability in the MTT assay, G1 arrest and significant apoptosis upon flow-cytometric analysis, presence of apoptotic bodies, formation of DNA ladders upon gel electrophoresis and inhibition of viral early gene expression by RT-PCR and Western blot. There was a good correlation between cell apoptosis and viral early gene inhibition after As2O3 treatment, suggesting that induction of apoptosis of HCE16/3 cells by As2O3 treatment might be associated with down-regulation of viral oncogene expression. In conclusion, our findings indicate that As2O3 induces apoptosis of HCE16/3 cells, which may provide a new approach for treating HPV-associated tumors.

[Studies on chemical constituents of the essential oil of Cyclorhiza waltonii].

Essential oil from the rhizome of Cyclorhiza waltonii was obtained by steam distillation. The chemical components of the oil were examined by means of capillary gas chromatography and gas chromatography-mass selective detector with HP-5 and Carbowax 20M stationary phases. Fifteen constituents of the oil were identified. The content of each component identified was determined by area normalization method. Of these, the content of ten compounds was higher than 2%. They are: alpha-pinene, sabinene, alpha-therpinene, p-cymene, beta-phellandrene, gamma-terpinene, terpinolene, terpineo-4-ol, cis-caryophyllene, duryl aldehyde and isoduryl aldehyde. The oil was found to compose of almost all the types of volatile composition, i.e., not only monoterpenes and sesquiterpenes but also alcohols and aldehydes. Duryl aldehyde and isoduryl aldehyde are not common in volatile oils.