Previously, we genetically engineered a Salmonella Typhi bacterial ghost (STG) as a novel inactivated vaccine candidate against typhoid fever. The underlying mechanism employed by the ghost in stimulating the adaptive immune response remains to be investigated. In this study, we aimed to evaluate the immunostimulatory effect of STG on mouse bone marrow-derived dendritic cells (BMDCs) and its activation of the adaptive immune response in vitro. Immature BMDCs were stimulated with STG, which efficiently stimulated maturation events in BMDCs, as indicated by upregulated expressions of CD40, CD80, and major histocompatibility complex class II molecules on CD11⁺ BMDCs. Immature BMDCs responded to STG stimulation by significantly increasing the expression of interleukin (IL)-6, which might indicate the induction of dendritic cell maturation in vivo (p < 0.05). In addition, ghost-stimulated murine BMDCs showed significant expressions of interferon gamma and IL-4, which can drive the development of Th1 and Th2 cells, respectively, in co-cultured CD4⁺ T cells in vitro. These results suggest that STG can effectively stimulate maturation of BMDCs and facilitate subsequent immune responses via potent immunomodulatory cytokine responses.
Adaptive Immunity ; Animals ; Bacteriophages ; Dendritic Cells ; Immunity, Innate ; In Vitro Techniques ; Interferons ; Interleukin-4 ; Interleukins ; Major Histocompatibility Complex ; Mice ; Salmonella typhi ; Salmonella ; T-Lymphocytes ; Th2 Cells ; Typhoid Fever

Sur8, a scaffold protein of the Ras pathway, interacts with Ras and Raf and modulates the Ras-extracellular signal-regulated kinase (ERK) pathway. Here we show that Sur8 is overexpressed in established human colorectal cancer (CRC) cell lines and CRC patient tissues. Moreover, Sur8 expression is increased during liver metastasis in CRC patients. Sur8 knockdown decreases ERK and Akt activities in CRC cell lines, regardless of their K-Ras, B-Raf or PI3K mutation status. Overexpression or knockdown of Sur8 increases or decreases, respectively, the proliferation or transformation of CRC cell lines. Sur8 knockdown attenuates the migration and invasion of HCT116 CRC cells. Subcutaneous or orthotopic injection of HCT116 cells harboring a doxycycline (Dox)-mediated Sur8 knockdown system in nude mice resulted in decreased tumorigenic potential and inhibited the liver metastatic potential of HCT116 cells. Taken together, our data support the role of Sur8 as a promoter of tumorigenesis and liver metastasis in CRC through its modulation of the Ras-ERK and PI3K-Akt signaling pathways.
Animals ; Carcinogenesis* ; Cell Line ; Colorectal Neoplasms* ; Doxycycline ; HCT116 Cells ; Humans ; Liver ; Mice ; Mice, Nude ; Neoplasm Metastasis* ; Phosphotransferases

In an effort to develop novel mushroom-derived anti-obesity nutraceuticals, water and ethanol extracts containing the lipaseinhibitory compound from Phellinus linteus were prepared, and their nutritional components were determined. The optimal conditions for the extraction of P. linteus lipase inhibitor involved the treatment of the fruiting bodies with distilled water at 80degrees C for 72 hr and 80% ethanol at 100degrees C for 60 hr, respectively. The distilled water extract and ethanol extract contained 10.9% and 6.11% of crude protein, and 0.96% and 15.86% of crude fat, respectively. Additionally, the distilled water extract contained a large quantity of minerals, including 239.5 mg of K, 39.3 mg of Mg, and 39.3 mg of Na. The free amino acid content of the distilled water extracts was also higher than that of the ethanol extracts, and in particular, the distilled water extracts contained 5,139 mg of asparagine, 3,891 mg of tryptophan, 2,598 mg of alanine, and 2,066 mg of serine in 100 g of the distilled water extracts. 100 g of the distilled water and ethanol extracts were found to contain 12.31 g and 8.16 g of malic acid, respectively.
Alanine ; Asparagine ; Dietary Supplements ; Ethanol ; Fruit ; Lipase ; Malates ; Minerals ; Serine ; Tryptophan ; Water

BACKGROUND/AIMS: The atrophic gastritis with intestinal metaplasia of gastric mucosa has been considered to be the major factor of carcinogenesis in the stomach. However, the key molecules are still poorly understood. To elucidate the molecular genetic basis, we report the results of our initial microarray data to analyze the genome pattern in patients with atrophic gastritis and intestinal metaplasia of the stomach. METHODS: We used oligonucleotide microarray technique to evaluate the gene expression profiles in atrophic gastritis with intestinal metaplasia, in comparison with those of normal mucosa. For the identification of differentially expressed genes, Significance Analysis of Microarrays (SAM) package method was used. The results were analyzed using global normalization, intensity dependent normalization, and box plot normalization. RESULTS: Eight genes including FABP, REG, OR6C1, MEP1, SLC6A1, SI, Mucin 1, and RAB23 in mucosa of atrophic gastritis and intestinal metaplasia were up-regulated by more than 10 times as compared with normal gastric mucosa. Only one gene, LOC44119 was down-regulated by more than 10 times of the expression as compared with normal gastric mucosa. In respect to the expression of known genes related to gastric carcinogenesis, 8 genes including FN1, SRMS, TP53, TP53IMP2, TP53I3, FGFR4, TGFB1, and TGFA showed up- and down-regulations more than 2 folds in expression pattern. CONCLUSIONS: We could identify a total genome pattern in patient with atrophic gastritis and intestinal metaplasia using oligonucleotide microarray. We believe that the current results will serve as a fundamental bioinformative basis for clinical applications in diagnosis and treatment of gastric cancer and precancerous lesion in the future.
Down-Regulation ; Gastritis, Atrophic/*genetics/metabolism ; Gene Expression Profiling ; Humans ; Intestines/*metabolism/*pathology ; Metaplasia/genetics/metabolism ; Microarray Analysis ; Tumor Markers, Biological/genetics/metabolism ; Up-Regulation

PURPOSE: A microarray-based gene expression analysis may offer a rapid and efficient means for assessing. However, the molecular genetic change in nonneoplastic colonic polyp is still poorly understood. To elucidate the molecular genetic basis, We now report the results of our initial microarray data to analyze the genom pattern in patients with hyperplastic polyps of colon. METHODS: 36 samples (18 pairs of colonic polyps and normal colonic mucosa were) harvested from colonoscopic biopsy. 3 of 18 colonic polyps were pathologically identified as the serrated type of hyperplastic polyp. We used the oligonucleotide microarray technique for analysis of the expression profiles of serrated polyps and normal mucosa. For the identification of differentially expressed genes, SAM (Significance Analysis of Microarray) package method was used. The result was analysed by using global normalization, intensity dependent normalization and block-wise normalization. RESULTS: Polypectomy specimens microscopically showed the pathologically characteristic serration with a saw-teeth like luminal border (branching of the crypts). 8 genes including RHEB (Ras homolog enriched in brain), WASF2 (WAS protein family, member 2), TYRP1 (Tyrosinase-related protein 1), VSX1 (Visual system homeobox 1 homolog), ROS1 (V-ros UR2 sarcoma virus oncogene homolog 1), WEE1 (WEE1 homolog), TEC (Tec protein tyrosine kinase), TNFRSF10A (Tumor necrosis factor receptor superfamily, member 10a) in serrated polyp were up-regulated by more than 10 times as compared with normal colonic mucosa. On the other hand, 6 genes including SIAT7D (Sialyltransferase 7D), DRD1 (Dopamine receptor D1), SIAT1 (Sialyltransferase 1), ITSN1 (Intersectin 1), TNFSF12 (Tumor necrosis factor superfamily, member 12), CHES1 (Checkpoint suppressor 1) were down-regulated by less than a tenth of the expression as compared with normal colonic mucosa. CONCLUSIONS: Serrated polyps as a subset of hyperplastic colonic polyps were analyzed with the oligonucleotide microarray technique. We authors could identify 14 genes (8 up-regulated and 6 down-regulated genes) that showed the significant change of expression as compared with normal colonic mucosa. Specifically, we believe that current study will serve as a fundamental base to offer a bioinformative characteristics of the serrated colonic polyp in future clinical applications.
Biopsy ; Colon* ; Colonic Polyps* ; Gene Expression Profiling* ; Gene Expression* ; Genes, Homeobox ; Hand ; Humans ; Molecular Biology ; Mucous Membrane ; Necrosis ; Oligonucleotide Array Sequence Analysis ; Oncogenes ; Phenobarbital ; Pilot Projects* ; Polyps ; Sarcoma ; Tyrosine

BACKGROUND: DNA vaccination represents an anticipated approach for the control of numerous infectious diseases. Used alone, however, DNA vaccine is weak immunogen inferior to viral vectors. In recent, heterologous prime-boost vaccination leads DNA vaccines to practical reality. METHODS: We assessed prime-boost immunization strategies with a DNA vaccine (minigene, gB498-505 DNA) and recombinant vaccinia virus (vvgB498- 505) expressing epitope gB498-505 (SSIEFARL) of CD8+ T cells specific for glycoprotein B (gB) of herpes simplex virus (HSV). Animals were immunized primarily with gB498-505 epitope-expressing DNA vaccine/recombinant vaccinia virus and boosted with alternative vaccine type expressing entire Ag. RESULTS: In prime-boost protocols using vvgBw (recombinant vaccinia virus expressing entire Ag) and vvgB498-505, CD8+ T cell-mediated immunity was induced maximally at both acute and memory stages if primed with vvgBw and boosted with vvgB498-505 as evaluated by CTL activity, intracellular IFN-staining, and MHC class I tetramer staining. Similarly gB498-505 DNA prime-gBw DNA (DNA vaccine expressing entire Ag) boost immunization elicited the strongest CD8+ T cell responses in protocols based on DNA vaccine. However, the level of CD8+ T cell-mediated immunity induced with prime-boost vaccination using DNA vaccine expressing epitope or entire Ag was inferior to those based on vvgBw and vvgB498-505. Of particular interest CD8+ T cell-mediated immunity was optimally induced when vvgB498-505 was used to prime and gB DNA was used as alternative boost. Especially CD8+ T cell responses induced by such protocol was longer lasted than other protocols. CONCLUSION: These facts direct to search for the effective strategy to induce optimal CD8+ T cell-mediated immunity against cancer and viral infection.
Animals ; Communicable Diseases ; DNA* ; Glycoproteins ; Immunity, Cellular* ; Immunization ; Memory ; Simplexvirus ; T-Lymphocytes ; Vaccination* ; Vaccines, DNA ; Vaccinia virus* ; Vaccinia*

Genomic DNAs extracted from 1,288 Haemaphysalis longicornis ticks collected from grass vegetation and various animals from nine provinces of Korea were subjected to screening by genus-specific (Ehrlichia spp. or Anaplasma spp.) real-time TaqMan PCR and speciesspecific (E. chaffeensis) nested-PCR based on amplification of 16S rRNA gene fragments. In all, 611 (47.4%) ticks tested positive for genus-specific amplification of 116 bp fragment of 16S rRNA of Ehrlichia spp. or Anaplasma spp. Subsequently, 396 bp E. chaffeensis-specific fragment of 16S rRNA was amplified from 4.2% (26/611) tick samples. The comparison of the nucleotide sequence of 16S rRNA gene from one tick (EC-PGHL, GeneBank accession number AY35042) with the sequences of 20 E. chaffeensis strains available in the database showed that EC-PGHL was 100% identical or similar to the Arkansas (AF416764), the Sapulpa (U60476) and the 91HE17 (U23503) strains. The phylogenetic analysis also revealed that the E. chaffeensis EC-PGHL formed a single cluster with the above strains. This is the first study to report molecular detection and phylogenetic analysis of E. chaffeensis from H. longicornis ticks in Korea. The implicit significance of E. chaffeensis infection in H. longicornis ticks in Korea is discussed.
Anaplasma/growth&development ; Animals ; Base Sequence ; Cloning, Molecular ; DNA, Bacterial/chemistry/genetics ; Ehrlichia chaffeensis/*genetics/growth&development ; Ehrlichiosis/*epidemiology/microbiology ; Korea/epidemiology ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Prevalence ; RNA, Ribosomal, 16S/chemistry/genetics ; Sequence Alignment ; Sequence Analysis, DNA ; Ticks/*microbiology

In the present study, we directly evaluated mucosal CD8+ T cell-mediated immunity using ex vivo cytotoxic T lymphocyte (CTL) assay and MHC class I tetramer staining method in iliac lymph node (LN) and vaginal tracts of mice immunized mucosally with several prime-boost protocols after genital infection of herpes simplex virus type 1 (HSV-1). Ex vivo CTL activity in iliac LN of infected mice was evaluated at 3-day post-infection without in vitro 5-day stimulation. Iliac LN of mice immunized with recombinant viral vaccine-priming and DNA vaccine-boosting protocol showed more potent CTL activity than those of other groups. Such ex vivo CTL activity was consistent with mucosal gB498-505 (SSIEFARL)-specific CD8+ T cell number of vaginal tract determined by MHC class I (H-2b) tetramer containing immunodominant peptide. Furthermore, the number of mucosal SSIEFARL-specific CD8+ T cells recruited into infected genital tracts appeared to decide the protective outcome against genital infection of virulent HSV-1. These results support that mucosal CD8+ T cells are principal mediators for the protection against genital infection of HSV-1.
Animals* ; Cell Count ; DNA ; Herpes Simplex* ; Herpesvirus 1, Human* ; Immunity, Cellular* ; Lymph Nodes ; Lymphocytes ; Mice ; Simplexvirus* ; T-Lymphocytes

PURPOSE: Pylorus-preserving pancreatoduodenectomy (PPPD) is an alternative surgical procedure for periampullary lesions. Early delayed gastric emptying is the most common and frustrating complication in the immediate postoperative period after PPPD and late delayed gastric emptying has been reported in some long-term follow-up studies. We evaluated the incidence of early delayed gastric emptying and analyzed temporal changes in gastrointestinal function after PPPD. METHODS: The incidence of early delayed gastric emptying was retrospectively evaluated from the medical records of 15 patients who underwent PPPD. Gastric emptying tests (GETs) using 99mTc-DTPA scan were performed on 11 of the patients every three months until 1 year, where possible. RESULTS: The incidence of early delayed gastric emptying was 6.7%. Five of the eight patients (62.5%) and six of the eight (75%) who underwent scintigraphy at 3 months and 6 months respectively, showed delayed gastric emptying. But at 12 months, all of the four patients who underwent GETs showed normal gastric emptyings. CONCLUSION: The incidence of early delayed gastric emptying after PPPD was 6.7%. Though there were few symptoms in long-term follow-up study using 99mTc-DTPA scan, delayed gastric emptying was frequently observed 3 to 9 months after PPPD. However, gastric emptying might be normalized in almost all patients around 1 year after PPPD.
Follow-Up Studies ; Gastric Emptying* ; Humans ; Incidence ; Medical Records ; Pancreaticoduodenectomy* ; Postoperative Period ; Radionuclide Imaging ; Retrospective Studies

The occurrence of carcinoma in a congenital uterine anomaly is uncommon. Indeed, malignancy of the uterine fundus with congenital uterine anomaly is quite rare, with fewer than 50 cases reported in the world liturature. Many patients go through life without the knowledge of their presence, and they are discovered at autopsy. Early diagnosis and proper management is necessary to decrease the high mortality. We present a case of endometrial carcinoma in a single horn of a bicornuate uterus.
Animals ; Autopsy ; Early Diagnosis ; Endometrial Neoplasms* ; Female ; Horns* ; Humans ; Mortality ; Uterus*