The epithelial cytokine response, associated with reactive oxygen species (ROS), is important in Helicobacter pylori (H. pylori)-induced inflammation. H. pylori induces the production of ROS, which may be involved in the activation of mitogen-activated protein kinases (MAPK), janus kinase/signal transducers and activators of transcription (Jak/Stat), and oxidant-sensitive transcription factor, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappaB), and thus, expression of interleukin-8 (IL-8) in gastric epithelial cells. alpha-lipoic acid, a naturally occurring thiol compound, is a potential antioxidant. It shows beneficial effects in treatment of oxidant-associated diseases including diabetes. The present study is purposed to investigate whether alpha-lipoic acid inhibits expression of inflammatory cytokine IL-8 by suppressing activation of MAPK, Jak/Stat, and NF-kappaB in H. pylori-infected gastric epithelial cells. Gastric epithelial AGS cells were pretreated with or without alpha-lipoic acid for 2 h and infected with H. pylori in a Korean isolate (HP99) at a ratio of 300:1. IL-8 mRNA expression was analyzed by RT-PCR analysis. IL-8 levels in the medium were determined by enzyme-linked immunosorbent assay. NF-kappaB-DNA binding activity was determined by electrophoretic mobility shift assay. Phospho-specific and total forms of MAPK and Jak/Stat were assessed by Western blot analysis. ROS levels were determined using dichlorofluorescein fluorescence. As a result, H. pylori induced increases in ROS levels, mRNA, and protein levels of IL-8, as well as the activation of MAPK [extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun NH2-terminal kinase 1/2 (JNK1/2), p38], Jak/Stat (Jak1/2, Stat3), and NF-kappaB in AGS cells, which was inhibited by alpha-lipoic acid. In conclusion, alpha-lipoic acid may be beneficial for prevention and/or treatment of H. pylori infection-associated gastric inflammation.
Enzyme-Linked Immunosorbent Assay ; Epithelial Cells/metabolism ; Gastric Mucosa/*drug effects/metabolism/microbiology ; Gene Expression Regulation, Bacterial ; Helicobacter Infections/immunology/*metabolism ; Helicobacter pylori/drug effects/*pathogenicity ; Humans ; Interleukin-8/genetics/*metabolism ; JNK Mitogen-Activated Protein Kinases ; Janus Kinase 1 ; Mitogen-Activated Protein Kinases/*biosynthesis ; NF-kappa B/*metabolism ; RNA, Messenger/isolation & purification/metabolism ; Reactive Oxygen Species/metabolism ; STAT3 Transcription Factor ; Stomach/metabolism/*microbiology ; Thioctic Acid/*pharmacology

Korea and Japan show the highest incidence of gastric cancer and Helicobacter pylori infection. New 2013 guidelines on H. pylori infection differ between the two countries with regard to the indications for H. pylori eradication, diagnostic methods, and treatment regimens. Indications for eradication in Korean guideline focus on specific diseases such as peptic ulcer disease, low-grade gastric mucosa-associated lymphoid tissue lymphoma, and after resection of early gastric cancer, while Japanese guideline includes all H. pylori-associated gastritis for the prevention of dissemination. With regard to the diagnosis, either noninvasive or invasive method (except for bacterial culture) is recommended in Korea, while two noninvasive tests including serum anti-H. pylori IgG antibody level are preferred in Japan. As for the treatment regimens, second-line treatment (quadruple bismuth-containing regimen) is recommended without first-line triple therapy in areas of high clarithromycin resistance in Korea. However, there is no bismuth-based second-line treatment in Japan, and the Japanese regimen consists of a lower dose of antibiotics for a shorter duration (7 days). Such discrepancies between the two countries are based not only on the differences in the literature search and interpretation, but also on the different approvals granted by the national health insurance system, manufacturing process of the antibiotics, and diagnostic techniques in each country. Collaborations are required to minimize the discrepancies between the two countries based on cost-effectiveness.
Anti-Bacterial Agents/pharmacology/*therapeutic use ; Antibodies/blood ; Bismuth/pharmacology/therapeutic use ; Disease Eradication/trends ; Drug Administration Schedule ; Guidelines as Topic ; Helicobacter Infections/complications/diagnosis/*drug therapy ; *Helicobacter pylori/drug effects/immunology ; Humans ; Japan ; Lymphoma, B-Cell, Marginal Zone/complications/surgery ; Republic of Korea ; Stomach Neoplasms/complications/surgery

OBJECTIVETo establish the chromatography-efficacy relation method for analyzing the anti-inflammatory activity of Qizhi Weitong particles, in order to lay a foundation for quality control and pharmacodynamic evaluation of traditional Chinese medicine compounds.

METHODOn the basis of a full-time multi-wavelength fusion fingerprint of Qizhi Weitong particles, the latin hypercube sampling was used to divide six herbs in Qizhi Weitong particles into groups of different proportions to determine their inhibition ratios of TNF-alpha, IL-6 and NO released by LPS-induced RAW264. 7 cells. Pharmaeodynamic data and chemical information of HPLC fingerprints of each group were analyzed with the gray correlation method to get the anti-inflammatory effect of each chromatographic peak, and then fitted with BP neural network to establish the chromatography-efficacy relation.

RESULTThere were 25 peaks closely related to the anti-inflammatory activity. With the 25 peaks as input items, the 3-BP network was adopted to establish the neural network model for anti-inflammatory effect of Qizhi Weitong particles.

CONCLUSIONWith an error of less than 7%, the model could better fit with the complicated non-linear relation of the compound, and applied in studying the chromatography-efficacy relation. In this study on the chromatography-efficacy relation, a new method is established to evaluate the anti-inflammatory activity of Qizhi Weitong particles. It is of practical significance as an effective approach for controlling quality and exploring the material basis for efficacy of traditional Chinese medicine compounds.

Animals ; Anti-Inflammatory Agents ; administration & dosage ; chemistry ; Cell Line ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Humans ; Interleukin-6 ; immunology ; Macrophages ; drug effects ; immunology ; Mice ; Neural Networks (Computer) ; Quality Control ; Rats ; Rats, Sprague-Dawley ; Stomach Diseases ; drug therapy ; immunology ; Tumor Necrosis Factor-alpha ; immunology

BACKGROUNDThe 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.

METHODSTILs 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.

RESULTSIL-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).

CONCLUSIONIL-18 and IL-12 can synergistically enhance cytotoxic functions of TILs from human gastric cancer.

Adult ; Aged ; Animals ; CD8-Positive T-Lymphocytes ; immunology ; metabolism ; Cell Line, Tumor ; Female ; Humans ; In Vitro Techniques ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-12 ; pharmacology ; Interleukin-18 ; pharmacology ; Interleukin-4 ; metabolism ; Lymphocytes, Tumor-Infiltrating ; drug effects ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Middle Aged ; Stomach Neoplasms ; immunology ; Tumor Necrosis Factor-alpha ; metabolism ; Xenograft Model Antitumor Assays

Inflammation is closely related to the progression of cancer as well as tumorigenesis. Here, we investigated the effect of prostaglandin E2 (PGE2) and interleukin-1beta (IL-1beta) on E-cadherin expression in SNU719 gastric cancer cells. E-cadherin expression decreased as the dose or exposure time of PGE2 and IL-1beta increased, whereas Snail expression increased with dose or time of PGE2 and IL-1beta. E-cadherin expression reduced by PGE2 treatment increased after the transfection of Snail siRNA. Neutralization of IL-1beta using anti-IL-1beta antibody blocked the expression pattern of E-cadherin and Snail occurred by IL-1beta treatment. However, there was no synergic effect of IL-1beta and PGE2 on the expression pattern of E-cadherin and Snail. In conclusion, inflammatory mediators reduced E-cadherin expression by enhancing Snail expression in gastric cancer cells. Inflammation-induced transcriptional regulation of E-cadherin in gastric cancer has implications for targeted chemoprevention and therapy.
Antibodies/immunology ; Antineoplastic Agents/pharmacology ; Cadherins/*metabolism ; Cell Line, Tumor ; Dinoprostone/*pharmacology ; Gene Expression Regulation/*drug effects ; Humans ; Interleukin-1beta/immunology/*pharmacology ; RNA Interference ; RNA, Small Interfering/metabolism ; Stomach Neoplasms/metabolism/pathology ; Transcription Factors/antagonists & inhibitors/genetics/*metabolism

OBJECTIVETo assess the efficacy and safety of compound matrine injection combined with cisplatin chemotherapy for advanced gastric cancer.

METHODIt was searched relevant randomized Controlled trials (RCTs) from Cochrane Library, PubMed, EMBASE, CBM, and CNKI etc. The search was finished in February 11, 2010. And it was traced the related references and experts in this field, besides it was also communicated with other authors in order to obtain some certain information that had not been found. RCTs of compound matrine injection combined with cisplatin chemotherapy versus cisplatin chemotherapy for advanced gastric cancer were included. It was evaluated the quality of these included studies and analyzed data by Cochrane Collaboration's RevMan 5.0 software.

RESULTTen RCTs were included meta analysis results suggested that compared with chemotherapy alone, the combination had a statistically significant benefit in healing efficacy (OR = 1.99, 95% CI: 1.26-3.13, P < 0.05) and improving quality of life (OR = 3.83, 95% CI: 2.38-6.15, P < 0.001). Besides, the combination also had a statistically significant benefit in myelosuppression, white blood cell (OR = 0.44, 95% CI: 0.32-0.62, P < 0.001), hematoblast (OR = 0.40, 95% CI: 0.26-0.60, P < 0.001), liver function (OR = 0.33, 95% CI: 0.15-0.75, P < 0.05) and in reducing the gastroenteric reaction (OR = 0.32, 95% CI: 0.16-0.63, P = 0.001), decreasing the of CD3 ( MD = 2.96, 95% CI: 1.724. 20, P < 0.001), CD4 (MD = 9.04, 95% CI: 7.87-10.20, P < 0.001), CD4/CD8 (MD = 0.47, 95% CI: 0.41-0.54, P < 0.001) and NK cells (MD = 5.90, 95% CI: 4.53-7.26, P < 0.001).

CONCLUSIONCompared with cisplatin chemotherapy, compound matrine injection combined with cisplatin chemotherapy can significantly improve the efficiency, QOL and myelosuppression, and reduce adverse events.

Alkaloids ; administration & dosage ; Bone Marrow ; drug effects ; Cisplatin ; administration & dosage ; Humans ; Injections ; Quality of Life ; Quinolizines ; administration & dosage ; Randomized Controlled Trials as Topic ; Stomach Neoplasms ; drug therapy ; immunology

OBJECTIVETo investigate the effect of Jianpi Yiqi Qingyou decoction on lymphocyte subsets and IL-2 mRNA in gastric tissue in rats with chronic superficial gastritis.

METHODWistar rats were randomly divided into 6 groups (11 for each): a blank control groups, the model of the control groups, the treatment groups (low-dose groups of traditional Chinese medicine, moderate-dose groups of traditional Chinese medicine, high-dose groups of traditional Chinese medicine) and lansoprazole groups. The models were made with the method in reference except a blank control groups. These rats are drinking freely with 0. 02% ammonia, continuous 90 days, and made preparations for experimental animal model of superficial gastritis. Making the model were detected by HE dying. The count of CD3+, CD4+ and CD8+ T cells were detected by immunohistochemistry. Using reverse transcriptase polymerase chain reaction (RT-PCR), the expression levels of IL-2 mRNA in gastric tissue were quantified.

RESULTCompared with that in model groups, the content of CD3+ T cells and CD4+ T cells in gastric tissue obviously increased in high dose of traditional Chinese medicine groups , the content of CD8+ T cells in gastric tissue obviously decreased in high dose of traditional Chinese medicine groups and the difference was significant (P < 0.01). The expression levels of IL-2 mRNA in gastric tissue obviously increased in moderate and high doses of traditional Chinese medicine groups, and the difference was significant compared with that in model group (P < 0.01).

CONCLUSIONJianpi Yiqi Qingyou decoction can obviously improve the content of CD3+ T cells and CD4+ T cells and the expression levels of IL-2 mRNA, decrease the content of CD8+ T cells in gastric tissue, improve immunity of rats. So the research results can provide some evidences for the treatment for chronic superficial gastritis.

Animals ; CD4-CD8 Ratio ; Chronic Disease ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gastritis ; genetics ; immunology ; pathology ; Interleukin-2 ; genetics ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach ; drug effects ; immunology ; metabolism ; pathology ; T-Lymphocyte Subsets ; drug effects ; metabolism

TNF-alpha is a major cytokine involved in inflammatory bowel disease (IBD). In this study, water extract of Grifola frondosa (GFW) was evaluated for its protective effects against colon inflammation through the modulation of TNF-alpha action. In coculture of HT-29 human colon cancer cells with U937 human monocytic cells, TNF-alpha-induced monocyte adhesion to HT-29 cells was significantly suppressed by GFW (10, 50, 100 microg/ml). The reduced adhesion by GFW correlated with the suppressed expression of MCP-1 and IL-8, the major IBD-associated chemokines. In addition, treatment with GFW significantly suppressed TNF-alpha-induced reactive oxygen species production and NF-kappaB transcriptional activity in HT-29 cells. In differentiated U937 monocytic cells, LPS-induced TNF-alpha production, which is known to be mediated through NF-kappaB activation, was significantly suppressed by GFW. In an in vivo rat model of IBD, oral administration of GFW for 5 days (1 g/kg per day) significantly inhibited the trinitrobenzene sulfonic acid (TNBS)-induced weight loss, colon ulceration, myeloperoxidase activity, and TNF-alpha expression in the colon tissue. Moreover, the effect of GFW was similar to that of intra-peritoneal injection of 5-aminosalicylic acid (5-ASA), an active metabolite of sulfasalazine, commonly used drug for the treatment of IBD. The results suggest that GFW ameliorates colon inflammation by suppressing production of TNF-alpha as well as its signaling through NF-kappaB leading to the expression of inflammatory chemokines, MCP-1 and IL-8. Taken together, the results strongly suggest GFW is a valuable medicinal food for IBD treatment, and thus may be used as an alternative medicine for IBD.
Animals ; Cell Adhesion/drug effects/immunology ; Cell Extracts/administration & dosage/*pharmacology ; Chemokine CCL2/biosynthesis/genetics ; Coculture Techniques ; Colon/drug effects/*metabolism/pathology ; Grifola ; HT29 Cells ; Humans ; Inflammatory Bowel Diseases/chemically induced/*drug therapy/pathology/physiopathology ; Interleukin-8/biosynthesis/genetics ; Intestinal Mucosa/*drug effects/metabolism/pathology ; Monocytes/*drug effects/metabolism/pathology ; NF-kappa B/genetics/metabolism ; Peroxidase/metabolism ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species/metabolism ; Stomach Ulcer ; Transcription, Genetic/drug effects ; Trinitrobenzenesulfonic Acid/administration & dosage ; Tumor Necrosis Factor-alpha/*biosynthesis/genetics ; U937 Cells ; Weight Loss

OBJECTIVEEGFR-mediated tumor proliferation plays an important role in the development of cancer, and is a key candidate for targeted therapy. The aim of this study is to evaluate the impact of EGFR monoclonal antibody Cetuximab (C225) on the growth, proliferation and apoptsis of gastric cancer xenograft in nude mice, and its possible mechanisms.

METHODSA gastric cancer cell line SGC-7901 with high EGFR expression level was screened from 7 gastric cancer cell lines. Gastric cancer xenografts in nude mice were established, and randomly divided into C225 treatment group and PBS control group. Tumor growth curves were calculated, the impact of C225 on the tumor growth, proliferation and angiogenesis was evaluated by immunohistochemical (IHC) staining Ki67 and CD34, respectively. The effect of C225 on apoptosis in the gastric cancer cells was evaluated by TUNEL assay. The expression levels of EGFR and its transcription factor Sp1 were detected by IHC staining and Western blot.

RESULTSAfter C225 treatment, the proliferation and growth of gastric cancer xenograft in nude mice were significantly decreased. In the contrast, the apopotic indexes in C225 treatment group and PBS control group were (16.4% +/- 0.3%) and (3.1% +/- 0.9%), respectively, with a significant difference (P < 0.001). There was no significant difference of the densities of CD34-positive microvessels between C225 treatment group and control group. Elevated expression of EGFR and Sp1 after C225 treatment was observed by IHC staining and Western blot assay.

CONCLUSIONEGFR monoclonal antibody cetuximab (C225) can effectively inhibit the growth of gastric cancer xenografts in nude mice, and trigger its apoptosis. Yet, C225 treatment may upregulate the expression of EGFR and its transcription factor Sp1. A "block-transcription activation-compensation" mechanism may exist to explain the molecular mechanism of acquired resistance of a single target blockade treatment.

Animals ; Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cetuximab ; Humans ; Ki-67 Antigen ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microvessels ; pathology ; Neovascularization, Pathologic ; prevention & control ; Random Allocation ; Receptor, Epidermal Growth Factor ; immunology ; metabolism ; Sp1 Transcription Factor ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; Tumor Burden ; drug effects ; Xenograft Model Antitumor Assays

OBJECTIVETo construct the murine CCL21 eukaryotic expression plasmid, and to investigate the chemotactic function of its products.

METHODSMurine CCL21 cDNA was amplified by RT-PCR from murine total RNA, and was inserted into eukaryotic expression plasmid pcDNA3.1 after confirmation of sequencing. The recombinant CCL21 plasmid was transferred into mouse forestomach carcinoma (MFC) cells and the chemotactic function of expressed products was detected by chemotaxis assay.

RESULTGene sequencing, gel electrophoresis of PCR products and restrictive digestion proved the successful construction of CCL21, and its expression was confirmed by Western Blot. The transfected tumor cells had a significant chemotactic function to DC.

CONCLUSIONThe recombinant murine CCL21 eukaryotic expression plasmid has been successfully constructed, and its expression products in tumor cells have a marked chemotactic function to DC.

Animals ; Base Sequence ; Chemokine CCL21 ; biosynthesis ; genetics ; Chemotaxis, Leukocyte ; Cloning, Molecular ; DNA, Complementary ; genetics ; Dendritic Cells ; drug effects ; immunology ; Genetic Vectors ; genetics ; Mice ; Mice, Inbred C57BL ; Molecular Sequence Data ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Stomach Neoplasms ; metabolism ; pathology ; Transfection ; Tumor Cells, Cultured