Latrodectus tredecimguttatus (commonly known as black widow spiders) have toxins not only in their venom glands, but also in other parts of their body, in their eggs and even in the newborn spiderlings. The study on the toxins in venom and materials outside the venom glands of the spiders to elucidate their differences and similarities, evolutional relationship and biological functions is of important theoretical and applicable significance. The development of modern protein chemistry and proteomics techniques has provided efficient means for the study of protein and peptide toxins of L. tredecimguttatus. By using such techniques, the molecular base and action mechanism of the toxins can be revealed at the levels of both single purified proteins and omics. Up to now, although protein chemistry and proteomics study on L. tredecimguttatus toxins have achieved a certain progress, the relevant work particularly that on the toxins in the materials outside the venom glands has to be further deepened.
Animals ; Arthropod Proteins ; chemistry ; Black Widow Spider ; chemistry ; Proteomics ; Venoms ; chemistry

Nasopharyngeal carcinoma (NPC) poses serious health problems in Southern China and yet the molecular mechanism of the carcinogenesis remains unclear. We used modern proteomic technologies to compare the protein expression profiles between the NPC cell lines (HNE1 and CNE1 ) and an immortalized nasopharyngeal epithelial cell line NP69 to identify cancer related proteins. Cell lysates were separated by two-dimensional gel electrophoresis (2 DE ) and analyzed by PDQuest software. The differentially expressed proteins were identified by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS). We discovered 15 up-regulated proteins and 18 down-regulated proteins in both HNE1 and CNE1 cell lines compared with NP69. These proteins are correlative with various functions, such as cell proliferation, apoptosis, cancer metastasis, metabolism, cytoskeleton and signal transduction. Western blotting analyses were further carried out to verify the differential expression of individual proteins. Several identified proteins in our research might be used as potential molecular markers to understand the molecular mechanism of NPC development and metastasis, and might be used as candidate targets for NPC treatments.

The neurotoxin peptide, hainantoxin-Ⅵ (HNTX- Ⅵ), has been isolated from the venom of Chinese tarantula Ornithoconus hainana by a combination of ion exchange chromatography and reverse phase HPLC. The toxin was found to contain 34 amino acid residues with 6 conserved cysteine residues. The effects of HNTX-VI on voltage-gated sodium channels were studied via whole-cell patch clamp techniques. Although several inhibitors of mammalian neuronal sodium channel activation (hainantoxin Ⅰ-Ⅴ) had been characterized from the same venom, the present study indicated that HNTX-Ⅵ had the ability to slow the inactivation kinetics of the sodium channels in Cockroach Periplaneta Americana dorsal unpaired median (DUM) neurons in a similar manner to δ-atractoxins. After HNTX-Ⅵ treatment, steady-state sodium channel inactivation became incomplete, leading to a non-inactivating component at potentials more positive than - 55 mV. The novel function of the tarantula toxin HNTX-Ⅵ not only supplies a useful tool for exploring the gating mechanisms of sodium channels but also provides theoretical foundations for exploiting novel and safe insecticides.

0.05). The mainly side-effects were myelosuppression, nausea, and vomiting. Conclusions:Docetaxel/cisplatin and gemcitabine/cisplatin regimens were well toleranced in advanced NSCLC patients with long term survival.

96% and with the same bio-activity as unlabeled Huwen toxin-1; Radioactivity detected in epidural space was 38% of injected radioacti vity at 10 min after epidural injection, which demonstrated successful administr ation into epidural space; The maximum serum concentration after epidural or iv administration of [ 125 I]labeled Huwentoxin-1 were determined to be (0 70?0 04) MBq?L -1 and (4 98?0 58) MBq?L -1 , respectively, a t the maximum serum concentration times of 30 min and 2 min. Terminal T 1/2 after epidural or iv administration were (10 36?0 27) h or (11 03?1 16) h, respectively. Cls was (1 29?0 07) L?h -1 ?kg -1 or (1 25? 0 23) L?h -1 ?kg -1 , respectively. Bioavailability after epidural a dministration was(95?5)%. CONCLUSION Concentration-time cur ves of [ 125 I] labeled Huwentoxin-1 after two routes were different. The degradation profiles after epidural and iv injection supported the using of HWTX-1 as analgesic by epidural administration.

Objective To examine the variation of protein expression in nasopharyngeal carcinoma cell lines with different biological characteristics and to identify the radiobiological associated proteins. Methods Biological characteristics of 5-8F and 6-10B were compared by flow cytometry assay after irradia- tion. The total proteins of 5-8F and 6-10B were separated by immobilized pH gradient(IPG) IEF-SDS two- dimensional gel electrophoresis technique. The differentially expressed proteins were cut from the gel and di- gested into peptides for MALDI-TOF MS and the Q-TOF mass spectrometric analysis. Identification of pro- tein was made through searching in protein sequence database. Protein expressions were examined by western blot and immunohistochemistry method. Results Nine most differentially expressed proteins between 5-8F cell and 6-10B cell were identified, p73 and CK19 expression examined by western blot were conformal with that by proteomic method, p73 expression in 5-8F cell was higher than in 6-10B cell. CK19 expression in 6- 10B cell was higher than in 5-8F cell. Conclusion Differentially expression of proteins exist in nasopha- ryngeal carcinoma cell lines with different biological characteristics. These proteins may be associated with cell radiobiological characteristic with the p73 as a potential biomarker.

Dorsal root ganglion (DRG) neurons are primary sensory neurons that conduct neuronal impulses related to pain, touch and temperature senses. To comprehensively identify proteins of plasma membrane (PM) from small amount of dorsal root ganglion (DRG) neurons, a proteomics strategy that utilizes aqueous polymer two-phase partition in combination with differential velocity centrifugation was adopted to enrich the PM, followed by SDS-PAGE, CapLC-MS/MS and bioinformatics analysis. Western blot analysis showed that the concentration of PM in purified plasma membrane(PPM) was 2.3 times higher than that in crude plasma membrane(CPM), 15 times higher than that in whole tissue lysate (WTL). By searching against the rat IPI protein sequence database, a total of 729 non-redundant proteins were identified from the PM preparation, of which 547 had a gene ontology (GO) annotation indicating a cellular component, and 159 (21.8%) were unambiguously identified as PM proteins. A data set of plasma membrane proteins of DRG as well as a tool to study PM proteins were provided in a small amounts of sample.

The antinociceptive effect of epidural administration of huwentoxin-I was elucidated in a tonic visceral pain rat model produced by acute colon inflammation. The nociceptive behaviors were induced by perendoscopically injecting dilute formalin (50 μl) into the depth of the colonic wall in rats. Both ω-conotoxinMVIIA and morphine hydrochloride were given epidurally as positive control while saline as negative control.Similar to ω-conotoxin-MVIIA and hydrochloride morphine, the epidural administration of HWTX-Ⅰ significantly reduced the nociceptive responses in a dose-dependent manner in tonic visceral pain rat model ( P ＜ 0.05). The suppression effects of both huwentoxin- Ⅰ and ω-conotoxin-MVIIA at 20 μg/kg were kept steady compared with the saline group and reached their maximum effects at the doses of 50 ～ 75 μg/kg within 1 hour when the nociception had been observed. It was also found that at the same doses, huwentoxin- Ⅰ was less effective in antinociception than ω-conotoxin-MVIIA. However, ω-conotoxin-MVIIA, but not huwentoxinⅠ , caused an obvious motor dysfunction at these doses. The action of morphine hydrochloride was initiated faster, but lasted for a shorter time than that of huwentoxin- Ⅰ and ω-conotoxin-MVIIA. Thus, huwentoxinⅠ , a potent blocker of neuronal N-type voltage-sensitive calcium channels, induced a remarkable dosedependent restrain effect similar to ω-conotoxin-MVIIA and morphine on the tonic visceral pain produced by colonic wall injection of formalin in conscious rats.

The low abundance and highly hydrophobic nature of most membrane proteins make their analysis more difficult than that for common soluble proteins. Successful membrane protein identification is largely dependent on the sample preparation including the enrichment and dissolution of the membrane proteins. A series of conventional and newly developed methods has been applied to the enrichment of low-abundance membrane proteins at membrane and/or protein levels and to the dissolution of hydrophobic membrane proteins. However, all the existing methods have inherent advantages and limitations. Up to now, there has been no unique method that can universally be employed to solve all the problems and more efforts are needed in improving sample preparation for the analysis of membrane proteomes.
Chromatography, High Pressure Liquid ; Mass Spectrometry ; Membrane Proteins ; analysis ; metabolism ; Membranes, Artificial ; Oxidation-Reduction ; Polyvinyls ; chemistry ; Proteome ; analysis ; Proteomics

A subcellular proteomic method was applied to investigate the protein expression profiles of nasopharyngeal carcinoma (NPC) cell lines,CNE1 and CNE2,at various differentiation levels.Plasma membrane (PM) proteins were obtained by Percoll density grade centrifugation and subjected to twodimensional electrophoresis (2DE) followed by PDQuest software analysis.Nine proteins expressed with more than two folds difference were identified by MALDI-TOF-TOF,of which functions involved in cell differentiation,signal transduction,and metabolism.Half of these proteins,such as galectin-1 and annexin Ⅱ,were analyzed with real-time quantitative PCR or Westem blotting.We have tested a proteomic method to study differentiated NPCs at different levels and found several proteins that might be related to their biological characteristics.