[Chinese multicenter randomized trial of customized chemotherapy based on BRCA1 (breast cancer susceptibility gene 1)-RAP80 (receptor-associated protein 80) mRNA expression in advanced non-small cell lung cancer (NSCLC) patients].

Objective: BRCA1 (breast cancer susceptibility gene 1) and RAP80 (receptor-associated protein 80) play key roles in predicting chemosensitivity of platinum and taxanes. A randomized trial was carried out to compare non-selected cisplatin-based chemotherapy with therapy customized according to BRCA1 and RAP80 expression. Methods: Advanced stage NSCLC patients whose tumor specimen was sufficient for molecular analysis were randomized (1∶3) to the control or experimental arm. Patients in the control arm received docetaxel/cisplatin; in the experimental arm, patients with low RAP80 expression received gemcitabine/cisplatin (Arm 1), those with intermediate/high RAP80 expression and low/intermediate BRCA1expression received docetaxel/cisplatin (Arm 2), and those with intermediate/high RAP80 expression and high BRCA1 expression received docetaxel alone (Arm 3). The primary end point was progression-free survival (PFS). Results: 226 patients were screened and 124 were randomized in this trial. ORR in the four subgroups was 22.6%, 48.4%, 30.3% and 19.2%, respectively (P=0.08); PFS was 4.74, 5.59, 3.78 and 2.73 months, respectively (P=0.55); and OS was 10.82, 14.44, 10.86 and 10.86 months, respectively (P=0.84). The common adverse effects included neutropenia, nausea, anemia and fatigue. Conclusions: No statistically significant difference of ORR, PFS or OS is observed in the experimental arms compared with the control arm. Patients with low RAP80 mRNA levels have a trend of better survival and higher response rate to gemcitabine/cisplatin chemotherapy.

RNA-electroporated CD40-activated B cells induce functional T-cell responses against HepG2 cells.

Hepatocellular carcinoma (HCC) is one of the incurable tumours in the world. Cell-based immunotherapy, in which antigen-loaded antigen-presenting cells (APCs) are able to elicit T cell responses, has become an alternative treatment for liver cancer. Here, we used HepG2 cells' total RNA-electroporated CD40 ligand-activated B (CD40-B) cells as alternative APC for induction of specific CD8+ T-cell responses. The antigen-presenting ability of CD40-B cells was determined by phenotypic analysis, showing a polyclonal, strongly activated B-cell population with high expression of co-stimulatory molecules. To demonstrate the ability of total RNA extracted from HepG2 cells electroporated CD40-B cells to induce CD8+ T-cell responses, these RNA-loaded cells were co-cultured with autologous peripheral blood mononuclear cells for 7 days followed by analysis of T-cell antigen specificity. These experiments showed that CD40-B cells electroporated with HepG2 cells' total RNA are capable of activating antigen-specific interferon-gamma-producing CD8+ T cells, and these T cells activated by CD40-B cells show a killing effect on HepG2 cells. These findings demonstrated that the carcinoma cell derived total RNA-electroporated CD40-B cells could be used as alternative APC for the induction of antigen-specific CD8+ T-cell responses, which might be used in HCC immunotherapy.

Sex differences in the neuroendocrine response to short-term fasting in rhesus macaques.

When energy intake is restricted in mammals, there are neuroendocrine adjustments in the secretion of reproductive and metabolic hormones to reallocate energy for vital functions. In the present study, we investigated whether there were differences in the luteinising hormone (LH), growth hormone (GH) and cortisol responses to a 48-h fast in adult gonad-intact male and female rhesus macaques. In both male and female macaques, blood glucose levels were significantly lower in fasted than in control studies, and levels were higher in males than in females. Male rhesus monkeys had significantly lower (P < 0.01) mean serum LH levels after a 48-h fast than under fed conditions and this was attributable primarily to a decrease in the amount of LH released during each secretory episode. In fasted females, serum LH levels were significantly greater (P < 0.05) than during the fed conditions but no differences were found in pulse amplitude or in the number of pulses. Almost twice as many GH pulses were observed in both males and females during fasting but there was no difference in either mean serum GH levels or pulse amplitude between control and fasted studies. A typical diurnal profile in cortisol levels was observed in both sexes and both experimental conditions. Under control conditions, male macaques released less cortisol than females, and although fasting increased mean cortisol levels in both males and females, only the males shown a significant rise over levels observed in control studies. The changes in plasma LH and cortisol levels in fasted rhesus macaques are similar to those observed in humans and suggest that gonadotrophin and corticotrophin secretion are more resistant to short-term energy deprivation in female than in male primates.

Differential chemotactic potential of mouse platelet basic protein for thymocyte subsets.

Mouse platelet basic protein (CXCL7/mPBP) was cloned from thymic stromal cells and further identification indicated that it was expressed in thymic monocytes/macrophages (Mo/Mphis). Recombinant mPBP was chemoattractive for target cells of polymorphonuclear leucocytes, peritoneal Mo/Mphis and splenic lymphocytes with distinct potencies. CXCR2 was identified to be a cognate receptor for mPBP. Mouse thymocyte subsets of CD4-CD8- double-negative (DN), CD4+CD8+ double-positive (DP), CD4+CD8- single-positive (CD4SP) and CD4-CD8+ single-positive (CD8SP) expressed cell surface CXCR2 with different positive percentages and expression levels. mPBP was chemoattractive for thymocyte subsets with the potency order DN>DP> CD8SP>CD4SP, consistent with the levels of CXCR2 expressed on the respective cells. Thus, mPBP in thymus is functionally redundant with chemokine CXCL12/ SDF-1. Moreover, our finding that thymic Mo/Mphis can produce mPBP implies that they may have other functions apart from acting as scavengers in thymus.

Identification of two novel CT antigens and their capacity to elicit antibody response in hepatocellular carcinoma patients.

FATE and TPTE genes were originally reported to be specifically expressed in the adult testis. We searched for the databases of Unigene and serial analysis of gene expression (SAGE) implying that these two gene transcripts might also be expressed in tumours. Herein, we demonstrated that FATE and TPTE mRNA transcripts were expressed in different histological types of tumours and normal testis. Both are cancer-testis (CT) antigens and renamed as FATE/BJ-HCC-2 and TPTE/BJ-HCC-5, respectively. Comparison at nucleotide sequence, the FATE/BJ-HCC-2 cDNA, was identical to that of FATE, whereas the TPTE/BJ-HCC-5 was found to have two isoforms in both cancers and testis: one was identical in cDNA sequence to TPTE, encoding a protein of 551 amino acids, and the other variant lacked an exon of 54 bp, encoding a protein of 533 amino acids. The mRNA expression was analysed by RT-PCR and real-time PCR. FATE/BJ-HCC-2 mRNA was detected in 66% (41 out of 62) in hepatocellular carcinoma (HCC) samples and 21% (three out of 14) in colon cancer samples, whereas the TPTE/BJ-HCC-5 mRNA was detected in 39% (24 out of 62) and 36% (five out of 14) in HCC and non-small lung cancer samples, respectively. The recombinant proteins were prepared and the reactivity of allogenic sera to these two antigens was screened. The frequency of antibody response against FATE/BJ-HCC-2 and TPTE/BJ-HCC-5 proteins was 7.3% (three out of 41) and 25.0% (six out of 24), respectively, in HCC patients bearing respective gene transcripts. Therefore, FATE/BJ-HCC-2 and TPTE/BJ-HCC-5 are the novel CT antigens capable of eliciting antibody response in cancer patients.

Heterogeneity within medullary-type TCRalphabeta(+)CD3(+)CD4(-)CD8(+) thymocytes in normal mouse thymus.

The functional maturation process of medullary-type CD4(-)CD8(+) [CD8 single-positive (SP)] thymocytes remains largely uncharacterized. We describe a phenotypic analysis of CD8 SP medullary-type thymocytes and find a remarkable heterogeneity within this thymic cell population. While mature CD8(+) T cells in the periphery are relatively homogeneous (TCRalphabeta(+)CD3(+)Qa-2(+) HSA(-)3G11(-)6C10(-)CD69(-)), CD8 SP medullary-type thymocytes contain discrete subpopulations that can be identified by differential expression of several cell-surface markers. We have identified at least six discrete subpopulations in the subset of TCRalphabeta(+)CD3(+) CD8 SP cells in the thymus. According to the expressed phenotypes, a linear developmental pathway is predicted among these CD8 SP subpopulations as follows: 6C10(+)CD69(+)HSA(hi)3G11(+)Qa-2(-) --> 6C10(-)CD69(+)HSA(hi/int)3G11(+)Qa-2(-) --> 6C10(-)CD69(-)HSA(int)3G11(+)Qa-2(-) --> 6C10(-)CD69(-)HSA(lo)3G11(+)Qa-2(-) --> 6C10(-)CD69(-)HSA(-/lo)3G11(-)Qa-2(-) --> 6C10(-)CD69(-)HSA(-/lo)3G11(-)Qa-2(+). This study provides a framework for understanding CD8 SP T cell maturation in the thymic medulla.

[Gas chromatographic determination of camphor, menthol, methyl salicylate, thymol in JEIL COOL PAP].

The contents of camphor, menthol, methyl salicylate and thymol in JEIL COOL PAP were determined with gas chromatography by using a stainless steel column (2 m x 3 mm i.d.) packed with 15% DEGS, Chromosorb W (AW-DMCS) 80-100 mesh. Temperature programming was from 70 degrees C to 180 degrees C. The quantitative determination was performed with diphenyl as the internal standard. The internal standard method showed good linearity (r = 0.9995-0.9999). The average recoveries were 99.63% (camphor), 99.83% (menthol), 100.0% (methyl salicylate) and 100.4% (thymol).

[Multiple types of cytokines secreted by a mouse fetal liver stromal cell line].

A murine fetal liver stromal cell line, called MFLC, has been established and maintained in DME supplemented with 10%NCS for 2 years. Without exogenous stimulation, the MFLC cells spontaneously secreted several types of cytokines, including IL-6, GM-CSF and chemotactic factor. Of which, IL-6 and chemotactic factor were abundant, GM-CSF at low level. Biological activities of IL-7 and IL-3 were not detected in the MFLC cell's supernatants. MFLC-SN induced colony formation of murine bone marrow cells in a dose dependent fashion in which mixed granulocyte/macrophage/megakaryocyte colonies(CFU-GMM) and granulocyte/macrophage colonies(CFU-GM) were dominant. MFLC-SN also sustained colony formation of bone marrow cells taken from 5-Fu injected mice, suggesting that there was a biological activity similar to SCF in MFLC-SN. The cytokines secreted by MFLC might play an important role in T cell early development in fetal liver. These results will be useful for us to analyse the mechanism of T cell development in fetal liver and to study the cytokine network regulation.