Antinociceptive effects of incarvillateine, a monoterpene alkaloid from Incarvillea sinensis, and possible involvement of the adenosine system.

Incarvillea sinensis is a Bignoniaceae plant used to treat rheumatism and relieve pain in traditional Chinese medicine. As a major component of I. sinensis, incarvillateine has shown analgesic activity in mice formalin tests. Using a series of animal models, this study further evaluated the effects of incarvillateine against acute, inflammatory, and neuropathic pain. Incarvillateine (10 or 20 mg/kg, i.p.) dose-dependently attenuated acetic acid-induced writhing, but did not affect thermal threshold in the hot plate test. In a Complete Freund's Adjuvant model, incarvillateine inhibited both thermal hyperalgesia and paw edema, and increased interleukin-1ß levels. Additionally, incarvillateine attenuated mechanical allodynia induced by spared nerve injury or paclitaxel, whereas normal mechanical sensation was not affected. Incarvillateine did not affect locomotor activity and time on the rotarod at analgesic doses, and no tolerance was observed after 7 consecutive daily doses. Moreover, incarvillateine-induced antinociception was attenuated by theophylline, 1,3-dipropyl-8-cyclopentylxanthine, and 3,7-dimethyl-1-propargylxanthine, but not naloxone, indicating that the effects of incarvillateine on chronic pain were related to the adenosine system, but not opioid system. These results indicate that incarvillateine is a novel analgesic compound that is effective against inflammatory and neuropathic pain, and that its effects are associated with activation of the adenosine system.

Inhibitory effects of the attenuated Salmonella typhimurium containing the IL-2 gene on hepatic tumors in mice.

To observe the inhibitory effects of an attenuated S. typhimurium strain carrying IL-2 gene (TPI) on hepatoma cell line (HepG2) and transplanted tumors in mice. TPI, TPG (an attenuated S. typhimurium strain carrying green fluorescent protein gene), and TP (an attenuated S. typhimurium strain) strains were transfected into HepG2 cells. At 48 h after transfecting, the transfection rate was 82.58 ± 1.74%. The expression level of IL-2 was (99.5 ± 12.2) ng/1 × 10(6) cells. Compared with TPG, TP, and normal mouse groups, the proportion of CD4(+) T and CD8(+) T cells in the blood from the TPI group was higher, the levels of IgM and IgG(1) were significantly increased, and the proliferation activity of splenic lymphocyte was significantly stronger. The transplanted tumor weight in the TPI group was significantly smaller than that in the other two groups. The infiltration of lymphocytes increased in the tumor from TPI group mice. TPI was effectively transfected into cancer cells, which expressed the protein of interest. Oral administration of TPI prolonged survival of mice transplanted with hepatoma cell tumours.

[Bortezomib combined with autologous peripheral blood hematopoietic stem cell transplantation for therapy of patients with multiple myeloma].

This study was aimed to evaluate the therapeutic efficacy of bortezomib combined with autologous peripheral blood hematopoietic stem cell transplantation (autoPBSCT) for patients with multiple myeloma (MM). 5 patients underwent autologous hematopoietic stem cell transplantation. Bortezomib treatment was supplied for patients before autoPBSCT and in the conditioning of transplantation, it was also used in maintaining treatment. Patients with transplantation adopted bortezomib plus melphalan conditioning regimen. The number of infused MNC and number of CD34(+) cells were 4.06×10(8) (4.09×10(8) - 4.37×10(8))/kg and 3.98×10(6) (2.49×10(6) - 8.2×10(6))/kg respectively. The results showed that hematopoiesis was reconstituted in 5 patients, with a neutrophil cell count more than 0.5×10(9)/L at day 14 (13 - 25 days) after transplantation and platelet count more than 50×10(9)/L at day 28 (21 - 41 days) after transplantation. Transplantation-associated death was not observed. 5 patients were disease-free survival. In conclusion, treatment of bortezomib combined with autologous peripheral hematopoietic stem cell transplantation is an effective method for patients with multiple myeloma. Use of bortezomib after transplantation might still be favourable to MM patients, for survival prolongation and life quality improvement.

[Application of Epstein-Barr virus-transformed B lymphoblastic cells in identification of CTL epitopes specific for Hantaan virus].

AIM: To establish Epstein-Barr virus (EBV)-transformed B lymphoblastic cell lines (B-LCL) to present peptides as antigen-presenting cells (APC) and stimulate short-cultured T cells secreting IFN-gamma, by which the T cell epitopes are identified. METHODS: PBMCs from patients with hemorrhagic fever with renal syndrome (HFRS) were transformed using EBV from supernatant of B95-8 cells. ELISPOT assay was then employed to evaluate the IFN-gamma production of short-cultured G9L-specific CD8(+) T cells stimulated with peptide-pulsed autologous B-LCL cells. RESULTS: B-LCL pulsed with G9L or G9L-nested V15R can stimulate G9L-specific CD8(+) T cells producing IFN-gamma, but not B-LCL pulsed with non-homologous I15P. CONCLUSION: B-LCL can efficiently and specifically present peptides to peptide-specific T cells as non-professional APC.

[Identification of HTNV-NP-specific T lymphocyte epitopes and analysis of the epitope-specific T cell response].

AIM: To identify the nucleocapsid protein of Hantaan virus (HTNV-NP)-specific T lymphocyte epitopes and analyze the epitope-specific T cell response during hemorrhagic fever with renal syndrome (HFRS). METHODS: T lymphocyte epitopes and frequencies of epitope-specific T cells were determined by ELISPOT using PBMCs from HFRS patients stimulated by individual or mixture of overlapping 15-mer peptides spanning the amino acid sequence of HTNV-NP. RESULTS: Out of 10 peptide mixtures, 8 elicited strong HTNV-NP-specific responses in 18 of 47 HFRS patients, and the T cell response was found at early stage of HFRS. Moreover, 17 HTNV-NP-specific T lymphocyte epitopes were identified in 11 patients, and most epitopes were clustered near the center of NP in linear structure. Among them, 14 T lymphocyte epitopes were described for the first time. CONCLUSION: HTNV-NP-specific T cell response can be elicited at early stage of HFRS and T lymphocyte epitopes mainly located in the center of NP, suggesting that it may play an important role in immune protection during HTNV infection.