Influence of FOLFOX regimen on the immunologic function in patients with advanced colorectal cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To examine the influence of chemotherapy with FOLFOX protocol (CT-F) on the immunologic function in patients with advanced colorectal cancer.</p><p><b>METHODS</b>A total of 43 patients with advanced colorectal cancer were included. Patients who received FOLFOX chemotherapy (Group A, n=22) were compared to those who did not(Group B, n=21). Blood was obtained from peripheral vein before chemotherapy (T0), at the time of completion of chemotherapy (T1), and 3 months after completion of chemotherapy (T2) to detect the percentage of regulatory T lymphocytes (CD4+CD25+Foxp3+T cells) in total T lymphocytes using fluorescence activated cell sorter (FACS). The level of Th1/Th2 cytokines in the serum including interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-10 (IL-10) and interferon-γ (IFN-γ) were detected by enzyme-linked immunoabsorbent assay(ELISA).</p><p><b>RESULTS</b>The percentage of regulative T lymphocyte was significantly lower at T1, which increased after chemotherapy but was still lower than that at T0 and that in Group B [(4.15±0.56)%, (5.60±0.88)%, and(5.38±0.92)%, all P<0.01]. The levels of IL-4, IL-10 decreased at T1, and increased to the normal level at T2 compared with those at T0 or those in the control group (all P<0.01). In contrast, the levels of IL-2 and IFN-γ increased significantly at T1 and decreased to the normal level at T2 during the entire observation period.</p><p><b>CONCLUSION</b>For patients with advanced colorectal cancer, FOLFOX chemotherapy can decrease the proportion of regulatory T lymphocytes and results in Th1/Th2 cytokine drift to Th1 type. Therefore, FOLFOX may help the release from the anticancer immune inhibition.</p>

Preparation of anticolon carcinoma vaccine with rich chaperone peptides and study on its anticancer efficacy / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy.</p><p><b>METHODS</b>CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50,000 and above 300,000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70,000, 90,000, 95,000, 110,000 and 170,000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL.</p><p><b>RESULTS</b>Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK(P<0.01).</p><p><b>CONCLUSIONS</b>The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42 centi-degree heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.</p>

Influence of chemotherapy with FOLFOX protocol on sex hormones of male patients and the protective effect of herbal medicines for reinforcing Shen and supplementing qi on it / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the influence of chemotherapy with FOLFOX protocol (CT-F) on sex hormones of male patients, and the protective and detoxifying effect of herbal medicines for reinforcing Shen and supplementing qi (HM) on it.</p><p><b>METHODS</b>The randomized block control and self-control design was adopted to retrospectively investigate the changes of sex hormones in 61 patients with cancer of colon, rectum or stomach. They were assigned to four groups, A: treated simply with HM; B: treated with CT-F; C: treated with CT-F combined with HM; D: the blank control group. One course of CT-F composed of 1 month treatment, and 6 courses totally were applied on each patients. The levels of luteinizing hormone (LH), estradiol (E2), prolactin (PRL), progesterone (P), testosterone (T) and follicle-stimulating hormone (FSH) were determined before treatment (T0), at the end of the 6th month treatment (T1) and the 12th month (T2) after starting treatment.</p><p><b>RESULTS</b>Levels of LH, PRL and T were significantly lowered in the group B at T1, being 4.6 +/- 0.4 IU/L, 8.6 +/- 0.7 microg/L and 13.1 +/- 1.4 IU/L, respectively, which were significantly different to those in the other 3 groups at the corresponding time; they were somewhat raised after chemotherapy but still lower at T2 than those at T0, being 5.0 +/- 0.4 IU/L, 9.9 +/- 1.1 microg/L and 14.1 +/- 1.4 IU/L respectively, also lower than the corresponding levels in the other 3 Groups (P<0.05 or P<0.01). In group C, LH significantly lowered at T1 (P<0.05) to 5.1 +/- 0.4 IU/L, but it was restored to the levels of T0 and that in Group D, reaching 6.1 +/- 0.5 IU/L; while PRL and T were changed insignificantly in the chemotherapeutic course, and restored at T2 to the level of T0, comparable to that in group D. Contrarily, levels of E2 and FSH increased significantly (P <0.01) in group B after chemotherapy (at T1) to 135 +/- 14 pmol/L and 9.1 +/- 1.1 IU/L respectively, and till T2, being 140 +/- 15 pmol/L and 9.1 +/- 1.o IU/L, they were lower than those at T0 and higher than those in group D, A and C ( all P <0.01), but the two indexes were not significantly increased in group C, being 116 +/- 12 pmol/L and 7.1 +/- 0.9 IU/L at T1. However, all the parameters showed no significant change in group A and D, and the level of P showed insignificant change in all the groups in the whole observation period.</p><p><b>CONCLUSION</b>CT-F could induce multiple sex hormonal abnormalities in male patients with post-operational gastrointestinal cancer, and HM shows protective and detoxifying effects on them in different degrees.</p>

Antitumour effect of total saponins of Rubus parvifolius on malignant melanoma / 中国中药杂志

<p><b>OBJECTIVE</b>To evaluate the antitumor effect of total saponins of R. parvifolius on malignant melanoma.</p><p><b>METHOD</b>The human malignant melanoma A375 cells were regularlly subcultured in vitro, and were divided into five groups contained positive control group (CTX), high concentration (0.01 mg x mL(-1)) and middle concentration (0.001 mg x mL(-1)) and low concentration (0.000 1 mg x mL(-1)) total saponins of R. parvifolius groups and negative control group. By using MTT colorimetric method, the cell viability was measured. B16 melanoma cells were transplanted to mice, which were divided into positive control group, high dose (100 mg x kg(-1)) and middle dose (50 mg x kg(-1)) and low dose (25 mg x kg(-1)) total saponins of R. parvifolius groups and negative control group. The inhibition effect of the tumor in vivo, mean survival time and rate of life-elongation of the mice were observed. TUNEL method was used to detect the apoptosis of B16 malignant melanoma.</p><p><b>RESULT</b>Antitumor assay in vitro showed that the absorbency increased in the concentration of 0.01, 0.001 mg x mL(-1) with statistical significance (P < 0.05 vs negative control). Antitumor assay in vivo showed that the tumor inhibitory rate of high dose (100 mg x kg(-1)) and middle dose (50 mg x kg(-1)) of total saponins of R. parvifolius were 37.02% and 30.61%, respectively. Loaded tumor mouse survival duration could be prolonged. The apoptosis indexes of B16 tumor cells in three treatment groups were 32.5%, 20.5% and 5.5%, respective and there was statistical significance (P < 0.05 vs negative control).</p><p><b>CONCLUSION</b>The total saponins of R. parvifolius has remarkable inhibition of proliferation of malignant melanoma cells in vivo and in vitro and exerts antitumor activities through promoting tumor cell apoptosis.</p>

Anticancer therapeutic effect of SEA-linked and membrane-bound HSP70-expressed intestine-carcinoma vaccine / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To develop a novel dual-modified vaccine, the superantigen-linked intestine-carcinoma cells expressing membrane-bound heat shock protein 70 (HSP70), and further examine its anticancer therapeutic effect.</p><p><b>METHODS</b>The pre-established intestine carcinoma CT26 line expressing membrane-bound heat shock protein 70 (HSP70) was amplified and incubated with superantigen fusion protein, staphylococcal enterotoxin A (SEA) fused with transmembrane sequence (SEA-TM), thereby the dual-modified vaccine was prepared after inactivation. The anticancer efficacy of the vaccine was examined.</p><p><b>RESULTS</b>The laser confocal microscopy and flow cytometry showed that there co-existed much HSP70 and SEA on the vaccine membrane surface. Both of the single-modified vaccines, the SEA-linked vaccine and membrane-bound-HSP70-expressing one, displayed marked tumor suppression, a prolonged survival period, augmented lymphocyte proliferation and higher NK and CTL activity in the vaccinated mice when compared with its counterpart. Furthermore, the dually modified vaccine induced lymphocyte proliferation most intensively, generated the highest NK and CTL activity as well as the strongest tumor rejection in the vaccinated mice. The survival period of the mice was further prolonged.</p><p><b>CONCLUSION</b>A new vaccine, SEA-linked and membrane-bound-HSP70-expressing intestine-carcinoma cells can induce more potent anticancer immunity and produce better therapeutic efficacy.</p>

Antitumor therapeutic effect induced by intestine-carcinoma cells vaccine expressing membrane-bound heat shock protein 70 / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To develop a new vaccine expressing membrane-bound heat shock protein 70 (mbHSP70) and further study its antitumor therapeutic effect.</p><p><b>METHOD</b>The pre- established vector expressing mbHSP70 was transfected into CT26 cells of colorectal cancer. After the CT26 cells were incubated with 900 microg/ml G418, the sub-clones resistant to G418 were harvested and the HSP70 positive clones were selected by limiting dilution. The clones were amplified and inactivated, thereby the vaccine expressing mbHSP70 was prepared. Lymphocyte proliferation stimulated by the vaccines, NK and CTL activity was observed. The antitumor efficacy of vaccine was observed in BALB/c mice model with colorectal cancer.</p><p><b>RESULTS</b>The laser confocal microscopy and flow cytometry showed that there existed much HSP70 on the vaccine membrane surface. The HSP70 gene-modified vaccine displayed augmented lymphocyte proliferation and higher NK and CTL activity in vitro,and marked tumor suppression and prolonged survival time of the vaccinated micein vivo, when compared with its counterpart. Furthermore, mbHSP70-expression vaccine elicited lymphocyte proliferation most intensively, generated the highest NK and CTL activity as well as the strongest antitumor effect, and prolonged survival time of the vaccinated mice.</p><p><b>CONCLUSION</b>A new vaccine expressing mbHSP70 has more potent antitumor immunity and better therapeutic efficacy than HSP70 gene-modified vaccine did.</p>