Interleukin-18 and -12 synergistically enhance cytotoxic functions of tumor-infiltrating lymphocytes / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The role of tumor-infiltrating lymphocytes (TILs) in the immunopathogenesis of individual cancer is not clear and is a challenge for anti-tumor immunotherapy. This study aimed to investigate the effects of interleukin (IL)-18 and -12 on cytotoxic functions of TILs.</p><p><b>METHODS</b>TILs from postoperative gastric cancer patients were costimulated with IL-18 and IL-12. SGC-7901 tumor cells were pre-incubated with TILs and subcutaneously injected into BALB/C SCID mice. The function of TILs was evaluated by measuring tumor sizes in tumor-bearing mice, T helper (Th)1 (tumor necrosis factor (TNF)-α, interferon (IFN)-γ) and Th2 cytokine levels (IL-10 and IL-4) in serum and cytotoxicity of mouse natural killer (NK) and CD8(+) T cells.</p><p><b>RESULTS</b>IL-18 and IL-12 synergistically inhibited the growth of SGC-7901 cells in vivo and significantly extended the survival rate of SGC-7901-bearing mice (66.7% vs. 13.7%, P < 0.01). Moreover, TILs could promote the secretion of TNF-α and IFN-γ ((130.34 ± 7.65) vs. (210.63 ± 12.31) pg/ml, P < 0.01; (14.23 ± 1.97) vs. (30.52 ± 2.12) pg/ml, P < 0.01), and downregulate IL-10 and IL-4 secretion ((103.72 ± 11.21) vs. (61.36 ± 5.41) pg/ml, P = 0.021; (49.36 ± 4.67) vs. (28.48 ± 3.86) pg/ml, P = 0.024).</p><p><b>CONCLUSION</b>IL-18 and IL-12 can synergistically enhance cytotoxic functions of TILs from human gastric cancer.</p>

Flow-dependent regulation of vascular function and gene expression in rat superior mesenteric artery / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Mesenteric artery thrombosis is prone to occur at specific arterial regions with different fluid flow patterns, yet mechanistic links between blood flow and vascular function remain unclear. This study aimd to investigate the role of blood flow in regulation of vascular function and gene expression in rats.</p><p><b>METHODS</b>Isometric tension was recorded in wire myograph to examine vascular function of specific regions (trunk parts and proximal parts from the origin) with different blood flow in superior mesenteric artery (SMA). Endothelial nitric oxide syntheses (eNOS), phosphorylated-eNOS (p-eNOS), serine-threonine kinase Akt and phosphorylated-Akt (p-Akt) protein expressions in SMA were examined by Western blotting. Significance was analyzed using a Student's t test or analysis of variance (ANOVA) followed by a Dunnett's multiple-comparison post hoc test.</p><p><b>RESULTS</b>Compared with trunks, proximal parts exhibited severely impaired relaxant responses to acetylcholine (Ach) (1 nmol/L to 10 micromol/L) (P < 0.01). p-eNOS and p-Akt protein levels were significantly reduced in proximal parts of SMA (0.37 +/- 0.03, 0.42 +/- 0.03 respectively) versus trunk parts (0.82 +/- 0.03, 0.72 +/- 0.03 respectively, both P < 0.05) while total eNOS and Akt expressions remain comparable in both regions by Western blotting analysis (0.70 +/- 0.03 vs 0.82 +/- 0.03; 0.70 +/- 0.03 vs 0.77 +/- 0.03 respectively, both P > 0.05).</p><p><b>CONCLUSION</b>Critical components that drive the vascular function and influence the localization of mesenteric artery thrombosis are flow-responsive elements within the vascular endothelium.</p>

Effects of anti-angiopoietin-2 antibody on vascularization of an implanted model of human colonic carcinoma on chick embryo / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To establish an implanted model of human colonic carcinoma on chick embryo, and to study the effects of anti-angiopoietin-2 antibody on its vascularization.</p><p><b>METHODS</b>The human colonic adenocarcinoma cell line HT-29 was transplanted on the chick embryo's chorioallantoic membrane(CAM), and the angiogenesis characteristics were observed by stero-microscope, light microscope and immunohistochemistry. Furthermore, the effects of anti-angiopoietin-2 antibody on angiogenesis and tumor growth were also investigated.</p><p><b>RESULTS</b>Three to seven days after HT-29 cell line was implanted into CAM, tumors grew rapidly and new blood vessels grew toward tumors. Five days after anti-angiopoietin-2 antibody was given, the number of blood vessels in anti-angiopoietin-2 antibody group was significantly down-regulated than that in tumor control group observed by stero-microscope (37.2+/-4.6 vs 56.8+/-7.4, P<0.01), but was up-regulated than that in normal control group (37.2+/-4.6 vs 9.6+/-2.4, P<0.01). Microvessel density(MVD) in anti-angiopoietin-2 antibody group was much lower than that in tumor control group by histological examination (9.6+/-2.4 vs 20.2+/-5.8, P<0.01).</p><p><b>CONCLUSION</b>Angiopoietin-2 antibody is able to inhibit the angiogenesis induced by colorectal cancer cell line HT-29 obviously. The anti-angiopoietin-2 antibody may be potentially useful for clinical treatment of colonic carcinoma.</p>

Effects of supernatant of cord blood CD3AK cells on proliferation, differentiation and apoptosis of HL-60 cells / 中华儿科杂志

<p><b>OBJECTIVE</b>The study was to investigate the impact of cord blood CD(3)AK cell culture supernatant (CS) on proliferation, differentiation and apoptosis of HL-60 cells.</p><p><b>METHODS</b>HL-60 cells were treated with different concentrations of CS (10%, 15%, 20%) for 3 days, 6 days and 9 days, and the same cells of control group were not treated with CS. The growth of induced cells was assessed with Trypan blue staining and cell counting with cytometer. The differentiation marker CD(11b) on the cell surface and cell-cycle was analyzed by flow cytometry (FCM), cell morphology (Wright-Giemsa staining) and NBT test to determine the extent of differentiation. Meanwhile, the changes of the apoptosis of the cells induced by 20% CS at different time points (3, 6 and 9 days) were analyzed by TUNNEL-POD, and the apoptotic characteristics of cells were observed.</p><p><b>RESULTS</b>The growth of HL-60 cell was inhibited as CS-inducing time and the dose of CS increased. At the same time, but HL-60 cell number in G(0)/G(1) phase of cell-cycle increased, but HL-60 cell number in S phase decreased compared with untreated group. The HL-60 cells induced by 20% CS for 9 days showed that (52.7 +/- 1.8)% of cells were at G(0)+G(1) phase and (43.8 +/- 1.1)% were at S phase (P < 0.05), which demonstrated that HL-60 cells induced by 20% CS underwent G(0)/G(1) phase cell-cycle arrest. The volume of the differentiated cells was enlarged gradually as CS-inducing time prolonged. After 3 days the differentiating cells began to express differentiating marker CD(11b) on the cell surface and the nuclei morphology of the differentiated cells was also changed and NBT-stained cells increased in number with the increased dose of CS increased. Three days after induction by 20% CS, the induced cells began to show signs of apoptosis and the apoptotic percentage of induced cells gradually increased with CS-induction time. The rate of apoptosis of cells was (33.3 +/- 2.3)% at 9 days (P < 0.01).</p><p><b>CONCLUSION</b>CS could not only inhibit the growth of HL-60 cells but also induce the differentiation and apoptosis in HL-60 cells.</p>

Expression of NFkappaB p65 and its target genes in gastric cancer and precancerous lesions / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the expression of NFkappaB p65 and its target genes in intestinal metaplasia (IM), dysplasia (Dys), gastric cancer (GC) infected with Helicobacter pylori (Hp) and explore the mechanism of infection by cytotoxin-associated antigen A expressing Hp (CagA(+)Hp) in the development of gastric cancer.</p><p><b>METHODS</b>CagA antibody in blood sample of 289 patients was determined by ELISA. Hp was detected by rapid urease test and Warthin starry staining. Expression of NFkappaB p65 and its target genes in IM, Dys and GC was examined by immunohistochemistry.</p><p><b>RESULTS</b>In IMI approximately II, IMIII, DysI, DysII approximately III and GC, the expression of NFkappaB p65 was significantly higher in patients with CagA(+)Hp infection than those without CagA Hp infection. In IMIII and DysII approximately III, the expression of NFkappaB p65, c-myc, CyclinD(1) and bcl-xl was significantly higher in patients with CagA Hp infection than those without CagA Hp infection. In gastric cancer infected with CagA(+)Hp, the expression of NFkappaB p65, c-myc, CyclinD(1) and bcl-xl was significantly higher in intestinal type than in diffuse type.</p><p><b>CONCLUSION</b>There are different mechanisms in intestinal type and diffuse type in the development of gastric cancer. The occurrence of intestinal type gastric cancer is associated with CagA(+)Hp infection which by NFkappaB p65 upregulating the expression of c-myc, CyclinD(1),bcl-xl in patients with IMIII, DysII approximately III. It may be an effective method to prevent gastric cancer by inhibiting NFkappaB p65.</p>

Effects of curcumin on the expression of cytokines in trinitrobenzene sulphonic acid induced colitis / 中华消化杂志

Objective To investigate the therapeutic effects of curcumin(Cur)on trinitrobenzene sulphonic acid(TNBS)induced colitis and to investigate cytokines change in colon mucosa,spleen and sera.Methods Colitis was induced in SD rats by intrarectal injection of TNBS(100 mg/kg).Experi- mental animals were divided into negative control group,TNBS group and Cur therapeutic group(Cur, 30 mg?kg~(-1)?d~(-1),intraperitoneal injection).Expression of cytokines mRNA in colon mucosa was observed by RT-PCR,intracellular cytokines of interferon(IFN)-?and interleukin(IL)-4 in splenocytes were detected by flow cytometry(FCM),concentrations of IFN-?and IL-4 in sera were determined by enzyme-link immunosorbent analysis(ELISA).Results After treatment with Cur,macroscopic scores (3.9?1.0 vs.2.2?0.7),myeloperoxidase(MPO)activity(15.0?2.6 vs.7.3?1.4),mRNA of IFN-?(1.02?0.07 vs.0.06?0.02,mRNAof IL-12(0.29?0.05 vs.0.11?0.01)and the ratio of IFN-?/IL-4(11.44?0.97 vs.0.38?0.10)in colon mucosa,proportion of IFN-?CD4~+(31.7?7.5 vs. 21.1?3.7)and the ratio of IFN-?/IL-4 CD4~+(19.9?5.1 vs.6.1?1.8)in splenocytes,concentra tions of IFN-?[(1528?159)pg/ml vs.(513?14)pg/ml] and the ratio of IFN-?/IL-4(19.5?4.1 vs. 4.2?0.6)in sera were all decreased(P＜0.05 or P＜0.01).Meanwhile,mRNA of IL-4(0.09?0.01 vs.0.15?0.04)and IL-10(0.28?0.08 vs.0.63?0.12)in colon mucosa,proportion of IL-4 CD4~+ (1.6?0.5 vs.3.4?1.1)in splenocytes and concentrations of IL-4 in sera[(81?15)pg/ml vs.(124?20) pg/ml] were all increased after the treatment of Cur(P＜0.05 or P＜0.01).Conclusions Cur showed ther- apeutic effects on TNBS-induced colitis,and the mechanism might be through regulating the balance of Th1/ Th2 in colon locally and systematically.