Hun-Soo Kim

In Vitro and In Vivo Antibacterial Activity of Punica granatum Peel Ethanol Extract against Salmonella.

Punica granatum is commonly used in Korea as a traditional medicine for the treatment of pathogenic bacteria. In this study, we investigated the in vitro and in vivo antimicrobial activity of P. granatum peel EtOH extract (PGPE) against 16 strains of Salmonella. The minimal inhibitory concentrations of PGPE were in the range of 62.5–1000u2009x03BCgu2009mL−1. In addition, the in vivo antibacterial activity of the PGPE extract was examined in a S. typhimurium infection mouse model. Mice were initially infected with S. typhimurium and then with PGPE. The extract was found to have significant effects on mortality and the numbers of viable S. typhimurium recovered from feces. Although clinical signs and histological damage were rarely observed in the treated mice, the untreated controls showed signs of lethargy and histological damage in the liver and spleen. Taken together, the results of this study indicate that PGPE has the potential to provide an effective treatment for salmonellosis.

TCL-SPION-enhanced MRI for the Detection of Lymph Node Metastasis in Murine Experimental Model

RATIONALE AND OBJECTIVESnThe aim of this study was to assess the feasibility of thermally cross-linked superparamagnetic iron oxide nanoparticle contrast (TCL-SPION) in magnetic resonance (MR) imaging (MRI) for the detection of lymph node metastasis in experimental model.nnnMATERIALS AND METHODSnB16F1 human melanoma cells were subcutaneously injected into the thighs of C57BL/6 mice (n = 10). MRI was performed 21 days after tumor injection using a 4.7-T MR scanner. In vivo MRI was performed before and after the intravenous administration of TCL-SPION using T2 fast spin-echo and T2 gradient-echo pulse sequences. Then, ex vivo MR images were obtained for resected inguinal lymph nodes (n = 18) using the same pulse sequences as for in vivo imaging. On the basis of hematoxylin and eosin staining results, the lymph nodes were classified into three groups: group 1, nonmetastatic; group 2, tumor volume <50% of the resected sample; and group 3, tumor volume >50% of the resected sample. Size, signal-to-background ratio, and enhancement pattern were evaluated in each of the three groups on ex vivo images.nnnRESULTSnThe findings observed on ex vivo MR images of 18 inguinal lymph nodes were compared with histopathologic findings. All nodes were classified into three groups: group 1, n = 6; group 2, n = 5; and group 3, n = 7. The sizes of the lymph nodes in group 1 were significantly different from the sizes of those in group 3 (P = .014), but there was no significant difference in lymph node sizes between groups 1 and 2 (P = .792). Signal-to-background ratios of samples in groups 2 and 3 were significantly higher than those of samples in group 1 (P = .045 and P = .007, respectively). Each group of lymph nodes showed characteristic enhancement patterns that were well correlated between the images and pathology, except for one node.nnnCONCLUSIONSnThe features and extent of metastasis in the lymph nodes corresponded to those observed on TCL-SPION-enhanced MR images. TCL-SPION-enhanced MRI is useful for the detection and estimation of lymph node metastasis.

MicroRNA 375 regulates proliferation and migration of colon cancer cells by suppressing the CTGF‐EGFR signaling pathway

MicroRNA 375 (MIR375) is significantly down regulated in human colorectal cancer (CRC) tissues; we have previously identified MIR375 as a colon cancer associated microRNA (miRNA). We identified putative MIR375 target genes by comparing the mRNA microarray analysis data of MIR375‐overexpressing cells with the candidate MIR375 target genes predicted by public bioinformatic tools. We investigated that the connective tissue growth factor (CTGF) is a direct target gene of MIR375. Expression of CTGF, a ligand of epidermal growth factor receptor (EGFR), was markedly enhanced in human CRC tissues in comparison with the corresponding normal colon tissues. We demonstrated that the expression levels of molecules in EGFR signaling pathways were regulated by MIR375 in colorectal cells. Using immunohistochemistry and the xenograft of MIR375‐overexpressing colorectal cells in mice, we showed that MIR375 regulates cell growth and proliferation, angiogenesis, cell migration, cell cycle arrest, apoptosis, and necrosis in colon cells. Furthermore, results of MIR375 overexpression and cetuximab treatment indicated that the apoptosis and necrosis in colon cells were synergistically enhanced. Our results suggest that the down‐regulation of MIR375 modulates EGFR signaling pathways in human colorectal cells and tissues by increasing CTGF expression; therefore, MIR375 may have a therapeutic value in relation to human CRC.

MicroRNA 429 Regulates Mucin Gene Expression and Secretion in Murine Model of Colitis

BACKGROUND AND AIMSnmiRNAs are non-coding RNAs that play important roles in the pathogenesis of human diseases by regulating target gene expression in specific cells or tissues. We aimed to detect miRNAs related to ulcerative colitis [UC], identify their target molecules, and analyse the correlation between the miRNAs and their target genes in colorectal cells and dextran sulphate sodium [DSS]-induced mouse colitis.nnnMETHODSnUC-associated miRNAs were identified by miRNA microarray analysis using DSS-induced colitis and normal colon tissues. The results were validated by quantitative real-time polymerase chain reaction [RT-PCR]. We identified target genes of MIR429, a colitis-associated miRNA, from our screen by comparing the mRNA microarray analysis in MIR429-overexpressed cells with predicted candidate target genes. We constructed luciferase reporter plasmids to confirm the effect of MIR429 on target gene expression. The protein expression of the target genes was measured by western blot,enzyme-linked immunosorbent assay [ELISA] analysis, or immunohistochemistry.nnnRESULTSnWe identified 37 DSS-induced colitis associated miRNAs. We investigated MIR429 that is down-regulated in DSS-induced colitis, and identified 41 target genes of MIR429. We show that the myristoylated alanine-rich protein kinase C substrate [MARCKS] is a direct target of MIR429. MARCKS mRNA and protein expression levels are down-regulated by MIR429, and MIR429 regulates the expression of MARCKS and MARCKS-mediated mucin secretion in colorectal cells and DSS-induced colitis. In addition, anti-MIR429 up-regulates MARCKS expression in colorectal cell lines.nnnCONCLUSIONnOur findings suggest that MIR429 modulates mucin secretion in human colorectal cells and mouse colitis tissues by up-regulating of MARCKS expression, thereby making MIR429 a candidate for anti-colitis therapy in human UC.

Adenolipoma of the thyroid gland: Report of a case with diagnosis by fine-needle aspiration cytology

Adenolipomas are rare benign neoplasms composed of mature adipose tissue and thyroid follicles. The preoperative fine‐needle aspiration cytology of such lesion has rarely been cited. Approximately 12 cases have been reported in the literature worldwide, diagnosed solely on histopathology. A 65‐year‐old woman presented with a 4‐month history of a thyroid nodule. A FNA cytology specimen showed a few benign follicular cells with adipose tissue. Right lobectomy was performed and the specimen revealed a solid yellowish mass measuring 3 × 2.5 cm. Microscopic findings showed a solid tumor predominantly composed of mature adipose tissue intermixed with thyroid follicles. The pathological diagnosis was adenolipoma of the thyroid gland. The presence of adipose tissue is a common finding within the cytologic specimen, especially in obese individuals or with inadequate sampling. But suspicion may be possible if excess amounts of adipose tissue are present in the submitted sample. Diagn. Cytopathol. 2008;36:253–256.

The Association of the GABRP Polymorphisms with Systemic Lupus Erythematosus

Gamma-aminobutyric acid receptor subunit pi (GABRP) is involved in inhibitory synaptic transmission in the central nervous system. This gene encodes multisubunit chloride channels and is also expressed in numerous nonneuronal tissues such as the uterus and the ovaries. This study was aimed to validate whether the polymorphisms in the GABRP gene are associated with the susceptibility to systematic lupus erythematosus (SLE). The genotype frequencies of the rs929763, rs732157, and rs3805455 of the GABRP gene in SLE patients were significantly different from those of the control group (P < 0.0001, P = 0.05 and 0.002, resp.). Additional analysis showed that the genotype of the rs929763 and rs3805455 of the GABRP gene were also significantly associated with female SLE patients (P < 0.0001, P = 0.005, resp.). Two haplotype frequencies including a major haplotype of GABRP SNPs were more significantly different between the SLE patients and the healthy controls (P = 0.038 and 4.2E − 24, resp.). These results suggest that the polymorphisms in the GABRP gene might be associated with the susceptibility to SLE and the haplotype of GABRP SNPs is useful genetic marker for SLE.

Identification of the Polymorphisms in IFITM2 and IFITM5 Genes and their Association with Ulcerative Colitis

Interferon inducible transmembrane protein (IFITM) family genes have been implicated in various cellular processes such as the homotypic cell adhesion functions of IFNs and cellular anti-proliferative activities. The present study aimed to investigate whether the polymorphisms of the IFITM2 and IFITM5 genes are associated with susceptibility to UC. We identified a total of thirteen polymorphisms (eleven SNPs and two variations) in the IFITM2 gene and twelve polymorphisms (eleven SNPs and one variation) in the IFITM5 gene, by the direct sequencing method. Genotype analysis in the IFITM2 and IFITM5 SNPs was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and Taq-Man probe analysis, and the haplotype frequencies of IFITM2 and IFITM5 SNPs for multiple loci were estimated using the expectation maximization (EM) algorithm. The genotype and allele frequencies of IFITM2 SNPs, as well as IFITM5 SNPs, in UC patients were not significantly different from those of the healthy controls. We also analyzed the combined frequencies of rs77537847 of IFITM1, rs909097 of IFITM2, and rs56069858 of IFITM5 in the UC patients and the healthy controls. Although the distribution of the major combined genotype frequency did not differ significantly between the healthy controls and the UC patients, the GGT combined frequency in the healthy controls was significantly different from that in the UC patients (P=0.002). This result suggests that the combined genotype of the IFITMs polymorphisms may be associated with a susceptibility to UC and could be a useful genetic marker for UC.

Human IL-27p28 Gene Polymorphisms are Associated with the Serum Total IgE Levels of Allergic Rhinitis Patients

Interleukin 27 (IL-27) was discovered as a heterodimeric cytokine of the IL-12 family, and is composed of two subunits - Epstein-Barr virus induced gene 3 (EBI3) and p28. It acts as a versatile cytokine in the early regulation of Th1 initiation and in the negative regulation of the Th2 factor GATA binding protein 3 (GATA-3). This cytokine is mediated by the IL-27 receptor (WSX-1), which is highly expressed on CD4? T lymphocytes and NK cells. We previously identified four polymorphisms in the human IL-27p28 gene and suggested that the polymorphism of IL-27p28 is associated with susceptibility to asthma. To determine whether these IL-27p28 SNPs are associated with susceptibility to allergic rhinitis, the genotype and allele frequencies of IL-27p28 SNPs were analyzed between allergic rhinitis patients and healthy controls. Although the genotype and allele frequencies of IL-27p28 SNPs in allergic rhinitis patients were not significantly different from those of the control group, there was a suggestive difference (P=0.037) between these groups in total serum IgE levels in the g.2905T＞G SNP of the IL-27p28 gene. Our result implies that the g.2905T＞G SNP of the IL-27p28 gene might have an affect on IgE production in allergic rhinitis patients.