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OBJECTIVE: To explore the effects of luteolin on the proliferation and migration of the endothelial cells co-cultured with cancer cells and its molecular mechanism. METHODS: Human umbilical vein endothelial cells (HUVECs) were primarily isolated and identified by the expression of VE-cadherin. Cancer-endothelial cell co-culture model was established using the Transwell system. The effects of luteolin at different concentrations (0 [Co-culture control], 20 and 50 μmol•L-1) on the proliferation and migration of HUVECs in the co-culture system were determined. A HUVECs control group removed of prostate cancer PC3 cells was also included. Human angiogenesis antibody array kit was used to assay the secretion levels of various protein factors in each group. RESULTS: VE-cadherin was expressed on all the cultured HUVECs. Increased proliferation ability was found in the HUVECs co-cultured with PC3 cells compared with that in HUVECs control group (P < 0.01). Treatment with 20 or 50 μmol•L-1 luteolin significantly inhibited the proliferation ability of the HUVECs in co-culture system (both P < 0.01). Increased migration ability was found in the HUVECs co-cultured with PC3 cells compared with that in HUVECs control group (P < 0.01). Treatment with 20 or 50 μmol• L-1 luteolin significantly inhibited the migration ability of the HUVECs in co-culture system (both P < 0.01). Secretion levels of multiple angiogenesis-related proteins in the cultural supernatant of co-culture system were significantly increased compared with those in HUVECs control group. Treatment with 20 μmol•L-1 luteolin significantly inhibited the secretion levels of interleukin-8 (IL-8), vascular endothelial growth factor (VEGF) and monocyte chemoattractant protein-1(MCP-1) in the cultural supernatant of co-culture system (all P < 0.01). CONCLUSION: Luteolin may inhibit the proliferation and migration of endothelial cells via suppressing the secretions of IL-8, VEGF and MCP-1 in cancer-endothelial co-culture system.

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OBJECTIVE: To investigate the cytotoxic activities of chemical constituents in alcohol extract of the stem bark of Murraya exotica L. METHODS: The cytotoxicity against five cancer cell lines, U937, HL-60, K562, Bel7402 and Hela, were assayed by MTT and SRB METHODS. The constituents were isolated from Murraya exotica L. by routine chromatographic METHODS and the structures of the isolates were elucidated by NMR techniques. The antitumor effect was evaluated on cancer cells in vitro. RESULTS: When the cancer cells were exposed to the extract for more than 48 h, the survival rate decreased with the increase of the drug concentration. Four compounds were isolated and identified as isolariciresinol (I), dimethoxy isolariciresinol (II), 3-methoxyisolariciresinol (III), and 5'-methoxyisolariciresinol (IV). CONCLUSION: The active ingredients in Murraya exotica L. have antitumor activities, which may be compounds I and II.

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Rice is highly sensitive to cold stress during reproductive developmental stages, and little is known about the mechanisms of cold responses in rice anther. Using the HiSeq™ 2000 sequencing platform, the anther transcriptome of photo thermo sensitive genic male sterile lines (PTGMS) rice Y58S and P64S (Pei'ai64S) were analyzed at the fertility sensitive stage under cold stress. Approximately 243 million clean reads were obtained from four libraries and aligned against the oryza indica genome and 1497 and 5652 differentially expressed genes (DEGs) were identified in P64S and Y58S, respectively. Both gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were conducted for these DEGs. Functional classification of DEGs was also carried out. The DEGs common to both genotypes were mainly involved in signal transduction, metabolism, transport, and transcriptional regulation. Most of the DEGs were unique for each comparison group. We observed that there were more differentially expressed MYB (Myeloblastosis) and zinc finger family transcription factors and signal transduction components such as calmodulin/calcium dependent protein kinases in the Y58S comparison group. It was also found that ribosome-related DEGs may play key roles in cold stress signal transduction. These results presented here would be particularly useful for further studies on investigating the molecular mechanisms of rice responses to cold stress.

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OBJECTIVE: To study the in vitro metabolism and enzyme kinetics of di-n-butyl-(4-chlorobenzohydroxamato) tin(IV) chloride in rat liver microsomes, and to identify the major cytochrome P450 isozymes involved in the metabolism of di-n-butyl-(4-chlorobenzohydroxamato) tin(IV) chloride in rat liver microsomes. METHODS: By optimizing the incubation conditions of di-n-butyl-(4-chlorobenzohydroxamato) tin(IV) chloride in rat liver microsomes, the enzyme kinetics in different enzyme sources was researched; the cytochrome P450 isozymes involved in metabolism of di-n-butyl-(4-chlorobenzohydroxamato) tin(IV) chloride were preliminarily explored through in vitro inhibition experiments. RESULTS: Different enzyme source metabolism experiments showed that between phenobarbital(PB) and desamethasone(Dex) induction groups and blank control group there had significant differences, but between the BNF group and blank control group there had no significant difference; the inhibition experiments revealed that ketoconazole had strong inhibition effect on di-n-butyl-(4-chlorobenzohydroxamato) tin(IV) chloride metabolism. CONCLUSION: CYP3A plays a leading role in di-n-butyl-(4-chlorobenzohydroxamato) tin(IV) chloride metabolism, and CYP2C9 may be partly involved. CYP1A has no catalysis action on metabolism of di-n-butyl-(4-chlorobenzohydroxamato) tin(IV) chloride. The Results suggest that attention should be paid to the possibility of drug interactions when di-n-butyl-(4-chlorobenzohydroxamato) tin(IV) chloride is combined with the drugs metabolized by above-mentioned isozymes.

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The genus bocavirus includes bovine parvovirus (BPV), minute virus of canines (MVC), and a group of human bocaviruses (HBoV1-4). Using sequence-independent single primer amplification (SISPA), a novel bocavirus group was discovered with high prevalence (12.59%) in piglet stool samples. Two nearly full-length genome sequences were obtained, which were approximately 5,100 nucleotides in length. Multiple alignments revealed that they share 28.7-56.8% DNA sequence identity with other members of Parvovirinae. Phylogenetic analyses indicated their closest neighbors were members of the genus bocavirus. The new viruses had a putative non-structural NP1 protein, which was unique to bocaviruses. They were provisionally named porcine bocavirus 1 and 2 (PBoV1, PBoV2). PBoV1 and PBoV2 shared 94.2% nucleotide identity in NS1 gene sequence, suggesting that they represented two different bocavirus species. Two additional samples (6V, 7V) were amplified for 2,407 bp and 2,434 bp products, respectively, including a partial NP1 gene and the complete VP1 gene; Phylogenetic analysis indicated that 6Vand 7V grouped with PBoV1 and PBoV2 in the genus of bocavirus, but were in the separate clusters. Like other parvoviruses, PBoV1, PBoV2, 6Vand 7V also contained a putative secretory phospholipase A(2) (sPLA(2)) motif in the VP1 unique region, with a conserved HDXXY motif in the catalytic center. The conserved motif YXGXF of the Ca(2+)-binding loop of sPLA2 identified in human bocavirus was also found in porcine bocavirus, which differs from the YXGXG motif carried by most other parvoviruses. The observation of PBoV and potentially other new bocavirus genus members may aid in molecular and functional characterization of the genus bocavirus.

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OBJECTIVE: To sequence the complete sequence of bocavirus I with sequence independent single primer amplification (SISPA-PCR). METHODS: To exclude the co-effection samples, all clinical samples of diarrhea cases were screened with special primers of rotavirus, astrovirus, adenovirus, calicivirus and bocavirus I. The virus were enriched through ultracentrifugation. Other nucleic acids, such as human and bacteria genomes, were degradated by DNase I and RNase. DNA of bocavirus was Amplificated with SISPA-PCR, then purificated, cloned and sequenced. The sequences were alighmented in nr with blastn and assembled with DNAstar. RESULTS: A 4834bp sequence of bocavirus I were assembled. CONCLUSION: SISPA-PCR is an economical and efficient technique for sequence a virus complete genome.

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