Genomic Medicine, Biomarkers, and Health Sciences - Articles in Press

Investigation of interleukin-6 in hepatitis B patients with high blood glucose levels - Corrected Proof

2012-05-17

Abstract: Hepatitis B virus plays an important role in persistent inflammation with necrosis of the liver tissue, with the result that some patients with hepatitis develop diabetes. The present study aims to understand the correlation between pro-inflammatory interleukin-6 (IL-6) expression and insulin resistance in hepatitis B patients. Forty-three patients with diabetes, 68 patients with hepatitis, and 68 controls without diabetes and hepatitis were included in this study. The concentrations of glycated hemoglobin, glucose, and insulin antibodies in patients with hepatitis B and diabetes were significantly higher than those in controls without hepatitis and diabetes (p < 0.001). The IL-6 concentrations in the patients with hepatitis B were significantly higher than those in the group with diabetes and the control group. The results suggest that an increase in IL-6 level in hepatitis B patients may lead to the occurrence of high blood glucose levels.

Pseudozyma spp. and Barnettozyma spp. effectively kill cancer cells in vitro - Corrected Proof

Ren-Yu Hu, Ching-Fu Lee, Hsiu-Chuan Chou — 2012-05-16

Abstract: Cancer is the overall leading cause of death in developed countries and also worldwide, and being able to exploit an effective anticancer drug is the aim of all cancer scientists. However, many of the synthetic drugs produced so far usually cause serious side effects, which reduces their therapeutic efficacy. Discovering new drugs or auxiliary therapies derived from natural products might thus provide a novel opportunity for cancer therapy. A recent study reported that some lethal toxins can maintain their activity after being injected into mice. We therefore used two Pseudozyma spp. and three Barnettozyma spp. to examine whether these killer yeasts can preserve their lethal effect on cancer cells under the physical environment (optimum pH, temperature and osmolality, supporting a living cell accomplishes to proliferate, metabolize, differentiate and survive). Our preliminary results showed that both Barnettozyma spp. and Pseudozyma spp. have stronger cytotoxicity against HepG2 than Chang’s liver cells. According to the results of two-dimensional difference gel electrophoresis (2D-DIGE), a total of 115 and 27 proteins differentially expressed by 1.5-fold or more were observed for HepG2 and Chang’s liver cells, respectively. Furthermore, we explored the mechanism involved in the effect of the lethal yeast filtrates on liver cancer cells using 2D-DIGE and mass spectrometry.

Development of mini-Sep for nucleated cell purification and isolation - Corrected Proof

Wan-Ju Kung, Ching-Chiang Lin, Shiu-Ru Lin — 2012-05-16

Abstract: Despite the availability of various methods that can isolate nucleated cells from blood or body fluids, these methods still require centrifugation and other purification devices to work. The main purpose of this study was to develop a simple cell purification and isolation device (mini-Sep) that does not require any centrifugation equipment to operate and can be used to isolate and concentrate nucleated cells from blood or body fluids for use in cell-related experiments. Mini-Sep works by using a syringe or pump to inject samples, thereby filtering out any impurities by removing residual nucleated cells and holding them in a containment space that was designed as part of the device. The results show that mini-Sep possesses the capability to isolate and concentrate nuclear cells, can be operated in simple and easy steps, and saves time typically spent on centrifugation. It can be easily adapted for use in various laboratories and places that lack laboratory equipment.

Analysis of hydroxyethyl valine in hospital sterilization workers - Corrected Proof

2012-05-14

Abstract: The formation of N-(2-hydroxyethyl) valine (HEV) in hemoglobin has been considered as a biologically effective dose for exposures to ethylene oxide (EO). In this study, 148 volunteers with no EO exposure history and 76 EO-exposed hospital sterilization workers in Taiwan were recruited, 10ml of blood was collected, and background information was gathered using questionnaires from each study subject. HEV was processed by following the modified Edman degradation method and quantitated using a gas chirography coupled with mass spectrometry. Statistical analysis shows that the formation of HEV was significantly associated with smoking status and EO exposure.

Proteomic analysis of quercetin-induced cardioprotective effects - Corrected Proof

Jing-Yi Chen, Hong-Lin Chan, Hsiu-Chuan Chou — 2012-05-14

Abstract: Cancer has continuously occupied the top leading cause of death in Taiwan for 29 years. Thus, scientists around the world actively devote themselves to the study of cancer therapeutics. Doxorubicin is one of the most efficient drugs in cancer therapy, but it also produces reactive oxygen species that induce serious cytotoxicity against heart cells. Quercetin, a plant-derived flavonoid, has been proven to contain potent antioxidant, antihistamine, antimicrobial, and anti-inflammatory properties. The aim of this study is to perform an in vitro investigation into whether quercetin is capable of decreasing doxorubicin-induced cytotoxicity and promoting the cell repair system in cardiomyocyte H9C2 cells. Proteomic analysis was performed to investigate the quercetin-induced responses.

Ruptured ovarian yolk sac tumor combined with hemoperitoneum in a young girl with abdominal blunt injury - Corrected Proof

2012-05-14

Abstract: We report our clinical experience concerning the incidental finding of a ruptured ovarian yolk sac tumor combined with hemoperitoneum in a patient with abdominal blunt injury. An girl 8 years of age visited emergency department due to vomiting and persistent abdominal pain after abdominal blunt injury. Physical examination revealed pale conjunctiva, abdominal distension, and tenderness over bilateral lower abdomen without any obvious peritoneal sign. After stabilization of her condition, one huge pelvic tumor with ascites was identified by abdominal sonography and computed tomography scan. Subsequently, hemoperitoneum was confirmed by paracentesis procedure. During exploratory laparotomy, one ruptured left ovarian tumor with bleeding was identified. Therefore, she underwent left oophorectomy and left salpingectomy. Typical structures of Schiller-Duval bodies and hyaline globules in the tumor cells were identified. Biopsy of the omental tissues also confirmed metastatic lesions of the tumor. Ovarian yolk sac tumor (FIGO Stage 3) was confirmed. She received four courses of chemotherapy after the operation and received subsequent regular follow-up at the hospital with an uneventful recovery. This case highlights the possibility of ruptured ovarian yolk sac tumor after abdominal blunt injury as the cause of internal bleeding, and it should be taken into consideration as one possible etiology of hemoperitoneum in a young girl.

Development of great burdock essence compounds - Corrected Proof

Ching-Hui Wang, Fu-Yen Chung, Tong-Rong Tsai, Shiu-Ru Lin, Hsueh-Chiao Liu — 2012-05-14

Abstract: The nanomicelle encapsulating the water-insoluble substances can solubilize in aqueous solution. In this study, we used nanomicelle technology to develop great burdock essence. Black semen oil extracts of Arnebia euchroma and Angelica sinensis, water extract of Arctium lappa, phospholipid, and glycerol were mixed and homogenized in a high-pressure homogenizer. The particle size of great burdock essence was 167.1±0.96nm. Concentrations of chlorogenic acid, shikonin, and ferulic acid of great burdock essence were 2.300±0.605, 1.484±0.009, and 1.463±0.006μg/g, respectively. The great burdock essence had nanoparticle-size molecules, which resulted in easy cellular uptake by endocytosis, and main components of three herbs. However, for more information on the health benefits of the great burdock essence, further studies are needed.

Internal transcribed spacer sequence analysis of Angelica from different habitats - Corrected Proof

Hsiao-Chun Hao, Jia-Yuan Chang, Fu-Yen Chung — 2012-05-14

Abstract: The internal transcribed spacer (ITS), located between the 18S and 26S nuclear ribosomal DNA (rDNA) sequences, has a high degree of variation. Analysis of ITS sequences is commonly used to identify the authenticity of Chinese herbal medicines. The aim of this study is to analyze ITS sequences of Angelica from different habitats to find out whether there are differences in sequence. Angelicas from three habitats were used in this study, including Taiwan, Sichuan Province (China), and Gansu Province (China). DNA was extracted from Angelicas, and ITS sequences were analyzed using polymerase chain reaction (PCR) and direct sequencing. The results showed that the similarity of ITS-1 and ITS-2 rDNA sequences in Angelicas produced in Gansu and Sichuan Provinces is up to 100%, and that produced in Taiwan and Sichuan Province is 88% and 87%, respectively. Therefore, we could use PCR and ITS sequencing analysis to identify whether these Angelicas were the same strain, in order to determine their worthiness and authenticity in terms of traditional Chinese medicine.

Molecular and genomic biomarkers for risk prediction of long-term end-stage liver diseases in patients with chronic viral hepatitis - Corrected Proof

Chien-Jen Chen — 2012-05-14

Abstract: Chronic hepatitis B and C virus (HBV; HBC) infections are worldwide public health challenges. Molecular and genomic biomarkers for the prediction of long-term risk of end-stage liver diseases in patients affected with chronic viral hepatitis are important for the clinical management of the diseases. The REVEAL-HBV/HCV study is a community-based prospective cohort study aimed to evaluate the risk predictors of progression of chronic HBV and HCV in Taiwan. A total of 23,820 participants were enrolled between 1991 and 1992 from seven townships in Taiwan. Their serum samples were collected at study entry and follow-up examinations and tested for antibodies against HCV (anti-HCV), HBV surface antigen (HBsAg) and e antigen (HBeAg), serum levels of alanine aminotransferase (ALT), HCV RNA, HBV DNA, HBsAg, and α-fetoprotein. Genotypes and mutant types of HBV and HCV in serum samples were also examined. Newly developed cases of cirrhosis and hepatocellular carcinoma (HCC) were ascertained through follow-up examination and data linkage with profiles from the National Cancer Registry, National Health Insurance Database, and Death Certification System. Age, sex, elevated serum levels of ALT, HBV DNA and HBsAg, HBeAg serostatus, HBV genotype, family HCC history, and alcohol drinking habits are important biomarkers for the prediction of HBV-related HCC and cirrhosis. Age, elevated serum levels of ALT and HCV RNA, and HCV genotype are important biomarkers for the prediction of HCV-related HCC. Risk calculators have been developed for the long-term risk prediction of HCC for patients with chronic HBV and HCV. These risk calculators have been documented to have good validity for the prediction of risk of HCC and cirrhosis.

Searching for cellular factors associated with HCV infection - Corrected Proof

Michael M.C. Lai — 2012-05-14

Abstract: Hepatitis C virus (HCV) infection is the leading cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma in many parts of the world. It is also often associated with steatosis and other alterations of lipid metabolism, such as hypocholesterolemia. To identify the potential biomarkers for HCV-associated diseases, we have used various approaches to identify the cellular factors involved in HCV replication and pathogenesis. These factors may serve as the biomarkers for various stages of the disease. We have approached this question by identifying cellular proteins coimmunoprecipitated with the viral proteins. We also screened for cellular factors by performing genome-wide or functional subset inhibitory RNA (RNAi) library screening. Many cellular proteins have been identified, among which are proteins belonging to the vesicular transport pathway. For example, vesicle-associated membrane protein (VAMP)-associated vesicular proteins A and B (VAP-A and -B) were found to be required for HCV replication. We also found that HCV RNA replication takes place on the lipid rafts in an induced double-membraned vesicle (termed the “membranous web”). Correspondingly, several cellular proteins involved in the vesicle formation and lipid raft rearrangement were found to be relocalized or induced in HCV-infected cells. Among these are proline-serine-threonine phosphatase-interacting protein-2, a membrane-deforming protein, annexin A2 (a lipid raft organizer), and phosphatidyl inositol phosphate-4, a membrane component. The identification of the essential roles of these proteins in HCV replication indicates that the intracellular membrane and lipid metabolism undergo drastic reorganization during HCV infection. These and the related proteins may serve as biomarkers for HCV infection and disease progression.

The use of multiple molecular markers as predictors of the clinical prognosis of patients with colorectal cancer - Corrected Proof

Ting-Chen Tung, Shiu-Ru Lin, Jaw-Yuan Wang, Fu-Yen Chung — 2012-05-14

Abstract: Serum carcinoembryonic antigen (CEA) is most commonly used as a prognostic biomarker for evaluating curatively resected colorectal cancer (CRC) patients, but it has a low sensitivity and specificity. The aim of this study was to evaluate potential genetic markers in CRC patients using membrane array. Fifty CRC patients were enrolled and mRNA expression in their tissues were analyzed using membrane array analysis. Seven genes were analyzed in this study, including ATP2A2, GLUT1, MMP13, MAGE-A2, MAGE-A7, MAGE-A8, and MAGE-A12. Correlations between the results of the membrane array and the clinicopathological features of these CRC patients were then evaluated. The results show that the overexpression of any three or four of these seven genes is correlated with tumor invasion depth, lymphatic invasion, advanced stage, and postoperative recurrence (all p < 0.005). Furthermore, the expression of any four genes was more significantly correlated with clinicopathological characteristics than the expression of only two or three genes. The combination of multiple molecular markers and the membrane array method might be useful for predicting postoperative relapse in CRC patients.

Combining MALDI-TOF and molecular imaging with principal component analysis for biomarker discovery and clinical diagnosis of cancer - Corrected Proof

Yi-Tzu Cho, Yi-Yan Chiang, Jentaie Shiea, Ming-Feng Hou — 2012-05-14

Abstract: Molecular imaging using matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF MS) is effective for determining the distribution of molecules of interest in specific tissues. It can determine the direct correlation between metabolite, lipid, and protein expression and histology. Principle component analysis (PCA) can reduce the dimensions of a data set while still retaining the information present in the original data set. Using PCA to process MALDI data, samples with different statuses and ion patterns on their MALDI mass spectra can be classified, grouped, and evaluated on the same score plot. The use of MALDI-TOF in combination with PCA to compare the lipid, peptide, and protein profiles of different biological specimens can then be used to diagnose disease. Because ions with significant differences between sampling regions in a tissue can be indicated using PCA, the imaging of these “interesting peaks” can be visualized by plotting the ion intensity across the tissue section.

Identification of colorectal cancer recurrence-related microRNAs - Corrected Proof

2012-05-11

Abstract: Colorectal cancer (CRC) has high recurrence rate.Effective biomarkers for the detection of colon cancer are still unavailable. MicroRNA (miRNA) is an epigenetic factor that regulates cell proliferation, tumor cell growth, cancer formation, and metastasis by regulating tumor suppressor genes or oncogenes. miRNA has the potential to be used as a biomarker for the diagnosis of diverse cancers. Previously, we revealed that miR-139-3 p is downregulated and miR-338-5 p is upregulated in recurrent CRC patients. The present study further reveals that the miR-139-3 p expression level is inversely correlated with increased metastatic status in three colon cancer cell lines (SW480, SW620, and colo201), as determined using real-time polymerase chain reaction. Furthermore, in 51 pairs of CRC clinical specimens, the expression level of mir-139-3 p was significantly lower in the tumor sections than in the adjacent normal tissues (p < 0.0001), indicating that they are tumor-suppressive miRNAs. Furthermore, the expression level of mir-338-5 p in the tumor tissues of metastatic patients was significantly higher than in the tumor tissues of nonmetastatic patients (p < 0.05), indicating that mir-338-5 p is positively correlated with metastasis. All together, mir-139-3 p and mir-338-5 p may have potential use as biomarkers for the diagnosis of tumor formation and metastasis in CRC patients.

DNA-dependent protein kinase regulated glioblastoma survival in doxorubicin-induced cytotoxicity - Corrected Proof

Li-Hsun Lin, Hong-Lin Chan, Hsiu-Chuan Chou — 2012-05-11

Abstract: Generally, chemotherapy is effective when the cancer cell is dividing; most drugs trigger cancer cells to undergo apoptosis by attacking the cell’s DNA. In the process of cancer cell apoptosis, cancer cells become more resistant to chemotherapy treatments over time. Since DNA-dependent protein kinase (DNA-PK) plays an important role in DNA repairing, it is interesting to investigate the relationship between this particular enzyme and the development of multidrug resistance. In this study, we chose the commonly used chemotherapy drug doxorubicin to treat glioblastoma cells (M059k and M059j), and performed 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenykterazolium bromide (MTT) assay and immunofluorescence staining to assess the presence of DNA-PK. The result of MTT assay showed that the concentration of an inhibitor/drug required to reduce the cell viability by half of M059j is 1.75μm while that of M059k is 0.71μm after doxorubicin treatment. Comparing the staining result of M059j and M059k, DNA-PK was more detectable in M059k than in M059j. It suggested that further experiments need to be performed to identify and characterize the proteins that are important for signal transduction pathways that actually link DNA-PK with doxorubicin-induced cytotoxicity as well as those that are drug resistant.

Proteomic analysis of mitochondrial proteins in doxorubicin-resistant cancer cells - Corrected Proof

Yi-Wen Lo, Hong-Lin Chan, Hsiu-Chuan Chou — 2012-05-11

Abstract: Human uterine cancer occurs most often in menopause women between the ages of 55 and 70 years. In the initial stage, the symptoms of uterus have an abnormal vaginal bleeding or abnormal discharge, and the common treatment is to remove the uterus or treat with chemotherapeutic drugs. For most type of cancers, doxorubicin is frequently prescribed for treating a wide range of various types of cancers, including hematologic malignancies, lymphoma, myeloma, sarcoma, and uterine cancer. However, most cancer cells will become drug insensitivity or resistance via unknown mechanisms. Since mitochondria play a crucial role in the induction apoptosis, it is intriguing to clarify whether doxorubicin-resistance is mediated through mitochondrial protection. In this study, a highly doxorubicin-resistant cell line has been established by continuously culturing the MES-SA/Dx5, a commercial doxorubicin-resistant human uterine cancer cell line, in the presence of doxorubicin for up to 2 years. Mitochondrial proteins from nondrug-resistant MES-SA, low resistant MES-SA/Dx5 (MES-SA/Dx5low), and high resistant MES-SA/Dx5 (MES-SA/Dx5high) were enriched, and mitochondrial proteome is characterized by two dimensional-differential gel electrophoresis and matrix-assisted laser desorption/ ionization time of flight mass spectrometer (MALDI-TOF MS).

Microfluidic chip with microweir structure for continuous sample separating and collecting applications - Corrected Proof

2012-05-11

Abstract: We aim to establish a microfluidic chip device capable for continuous sample separation by integrating a simple and easy-fabrication microweir structure for crossflow filtration in the present study. The proposed microchip device is composed of two major components, including flow channels and microweir structure. Using the injection of the mixed samples with different sizes, the samples can be transported through the flow channel and then be separated by the microweir structure. The microweir structure with a different height can be generated by utilizing a standard lithography and overetching process, so that the gap can be used to be a selective tool to separate the smaller sample. In this study, a 10-μm gap was generated by established a microweir structure with 20 μm in height with a 20-minute etching process. Optimal sample separation efficiency in 82% can be obtained of the sample concentration in 103 μL−1 by utilizing the proposed design of the microweir structure to separate two groups of beads in different diameters. In conclusion, the proposed chip device can be regarded as an effective tool for clinical application.

Significance of migration-related genes (S100A9, MAGED4, C8orf30A, IL-8) in esophageal squamous cell carcinoma - Corrected Proof

2012-05-10

Abstract: To identify any biomarkers related to the migration of esophageal squamous cell carcinoma (ESCC) cells, ESCC CE81T cells were used to establish the CE81T-1 subline, which demonstrates increased migration activity after Transwell screening and microarray analysis. Among the differentially expressed genes, S100A9 was most downregulated, and MAGED4, C8orf30A, and IL-8 were the most upregulated in CE81T-1 cells. The expression of these four genes at the mRNA level was validated using the ESCC CE81T and KYSE cell lines and clarified in ESCC specimens using real-time polymerase chain reaction. Among 60 pairs of ESCC specimens (normal and tumor specimens), the expression level of S100A9 mRNA was significantly lower in the tumor sections in comparison with the normal sections (p=0.0228). In contrast, the expression level of IL-8 mRNA was significantly higher in the tumor sections in comparison with the normal sections (p=0.0061). Furthermore, C8orf30A expression was significantly correlated with ESCC metastatic status (p=0.0358) and associated with poorer survival (p=0.036), as determined by Kaplan-Meier analysis. Functional studies revealed that S100A9 plays a suppressive role in the proliferation and migration of ESCC cells through the overexpression of ectopic S100A9 and small interfering RNA, as determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Transwell assays. Altogether, this study reveals that C8orf30A has the potential to be used as a novel biomarker for the prognosis for ESCC metastasis and survival. Furthermore, the IL-8 and S100A9 genes may have potential in ESCC diagnosis.

Apoptotic-related gene expression in oxaliplatin-treated CCM-1 colorectal adenocarcinoma and MALT-nt-1 mucosa-associated lymphoid tissue lymphoma cell lines - Corrected Proof

Guan-Cheng Huang, Hsiao-Sheng Liu — 2012-05-10

Abstract: The purpose of this study was to assess the apoptotic -related gene expression of single-agent oxaliplatin in vitro. Growth inhibition studies were performed using the human cerebral cavernous malformation-1 colorectal adenocarcinoma cell line and mucosa-associated lymphoid tissue-nt-1 lymphoma cell line. At 50% inhibitory concentrations, oxaliplatin displayed suppressive effects toward Bcl-2 and the surviving expression in human cancer cell lines, whereas the expression of Fas/FasL was completely abolished in peripheral blood cells. Our data showed that oxaliplatin exerts potent antiapoptotic effects in human normal peripheral blood mononuclear cells at the suprapharmacologic concentration, warranting further investigations in the role of autoimmune adverse events while being used in numerous cycles for the treatment of malignancies.

Multiple mRNA markers for the detection of circulating tumor cells in breast cancer patients - Corrected Proof

Hsu-Chin Hung, Li-Chen Yen, Shiu-Ru Lin, Jaw-Yuan Wang — 2012-05-10

Abstract: The detection of circulating tumor cells (CTCs) is believed to be a potential method for the early detection of cancer. The aim of this study was to employ a membrane array to develop a panel of mRNA markers for the detection of CTCs in breast cancer patients. Twenty genes with highly differentiated expression levels were selected from microarray studies. The subsequent validation analysis of 30 pairs of breast tissue samples showed that 19 genes demonstrated two-fold overexpression in cancer tissues in comparison with those in normal tissues in 80% of the paired samples. Furthermore, a membrane array experiment was conducted on blood samples from 64 normal controls and 87 breast cancer patients, and the predictive power of each gene was evaluated by analyzing the gene expression level using receiver-operating curves (ROC). Among the 20 genes selected, eight genes that demonstrated the largest areas under the curve (AUC) > 0.8 were selected as diagnostic markers. According to the ROC analysis, when setting the cut-off point of five positive genes, the genetic array of the eight markers was recognized as positive with sensitivity and specificity values of 90% and 89%, respectively. These results suggest that this mRNA marker array could be useful for detecting CTCs in breast cancer patients.

Identification of DNA methylation biomarkers in imatinib-resistant chronic myeloid leukemia cells - Corrected Proof

2012-05-08

Abstract: Drug resistance is an obstacle to successful cancer treatment. In hematological neoplasms, such as chronic myelogenous leukemia (CML), drug resistance is often associated with gene hypermethylation and loss of function. An imatinib-resistant subclone of CML-like K562 (K562-R) was used as a model to study the role of hypermethylation in drug resistance. K562-R was selected by culturing cells with increasing concentrations of imatinib, ranging from 0.2–5 μM. A DNA methylation microarray was performed on the K562-R and parental K562 cells, and selected gene expression levels were confirmed using real-time polymerase chain reaction. The methylation level was significantly increased and the gene expression level significantly was decreased for MLH1, RPRM, FEM1B, and THAP2 in K562-R cells compared with parental K562 cells. Exposing K562-R cells to methylation inhibitors, such as 5-azacytidine (AzaC) and trichostatin A (TSA), reduced imatinib resistance. Our approach of using a drug-limiting dilution model followed by the use of a methylation microarray was able to identify methylation biomarkers for drug resistance. Specifically, MLH1, RPRM, FEM1B, and THAP2 might be potential epigenetic targets of imatinib resistance. Further understanding the methylation domain and epigenetic regulation machinery of these biomarkers will help researchers find potential effective therapeutic strategies that could be used to overcome drug resistance in CML patients.

Detection of type II diabetes mellitus using salivary transcriptomic biomarkers - Corrected Proof

Yu-Hsiang Lee, Kaumudi Joshipura, Jose Luis Vergara, David T. Wong — 2012-05-07

Abstract: Type II diabetes mellitus (T2DM) is one of the most common underdiagnosed metabolic diseases due to lack of recognizable symptoms in the early stage. T2DM can be largely prevented or controlled by diet or regular exercise at early stages, but often goes undetected for years, causing high rates of complications and mortality. Hence, a valid noninvasive early detection approach is urgently needed. In this study, we explored noninvasive detection of T2DM by salivary transcriptomic diagnostics. Salivary mRNA biomarkers were discovered by comparing microarray profiles of salivary transcriptomes in 13 T2DM patients and 13 healthy controls. The marker candidates selected from the microarray analysis were then subjected to verification in the original 26 samples using reverse transcription quantitative real-time polymerase chain reaction. Four up-regulated and two down-regulated mRNA biomarkers were validated. The logistic regression model showed that the combination of four identified biomarkers (KRAS, SAT1, EGFR, and PSMB2) could significantly distinguish T2DM patients from the healthy controls, yielding a receiver-operating characteristic-plot area-under-the-curve value of 0.917 with 100% sensitivity and 77% specificity. In conclusion, RNA signatures in saliva could serve as biomarkers for the detection of T2DM with high sensitivity and specificity, and offer a feasible means for early T2DM detection.

Investigation of vascular invasion and genetic polymorphisms of DPD IVS14+1 G>A, UGT1A1 3156 G>A, and UGT1A1 28 tandem repeats in colorectal cancer patients in Taiwan - Corrected Proof

Ming-Yii Huang, Meng-Lin Huang, Jaw-Yuan Wang, Shiu-Ru Lin — 2012-05-07

Abstract: In the present study, multiple chemotherapeutic agent-related genetic polymorphisms, including dihydropyrimidine dehydrogenase (DPD) IVS14+1 G>A, UGT1A1 3156 G>A, and UDP-glucuronosyltransferase (UGT)1A1 28 tandem repeats, were analyzed in patients with colorectal cancer and studied in correlation with the clinical features of those patients. The genotypes from 273 patients with stages I–IV colorectal cancer who underwent operations were determined by means of polymerase chain reaction-restriction fragment length polymorphism. The results showed that the genotype distribution of DPD GG, UGT1A1 3156 GG, and UGT1A1 28 tandem repeats 5/6 or 6/6 in Taiwanese subjects were 98.4%, 82.2%, and 80.6%, respectively. After analysis of the relationship between the genotypes and clinicopathological data of the patients, a significant correlation was observed between vascular invasion in patients with genetic polymorphisms ofDPDGG, UGT1A13156GG, and UGT1A1 28 repeat 5/6 or 6/6 (OR: 2.236, p = 0.015). There was a statistical correlation between vascular invasion and tumor invasion, lymph node metastasis, cancer stage, differentiation, perineural invasion, and survival (all p < 0.05). The results of the present study highly suggest that DPDGG, UGT1A13156GG, and UGT1A1 28 repeat 5/6 or 6/6 genotypes and vascular invasion could be prognostic factors for Taiwanese patients with colorectal cancer.

Microarray analysis and establishment of drug screening platform using 5-fluorouracil resistance HCT116 colon cancer cells - Corrected Proof

2012-05-07

Abstract: A systemic approach was used to identify the possible mechanisms underlying the development of 5-fluorouracil (5FU)-induced resistance on HCT116 colon cancer cells. From microarray analysis, HCT116 high-dose 5FU-resistant subclones showed differential gene expression compared to HCT116-sensitive clones. According to gene ontology, and Kyoto Encyclopedia of Genes and Genomes pathways, the up-regulated genes were related to cell death and lupus erythematosus, respectively. On the other hand, the down-regulated genes were related to cell division or DNA replication. Connectivity map (cMAP) analysis revealed that the molecular drugs, such as antiasthmatic or antiallergy agents that have negative correlations with cMAP score, may have beneficial effect for the resistant subclones. Our findings suggested that the feasibility of cMAP combining microarray gene expression profile may help identify a potential drug that possibly will reverse the effect of 5FU-induced resistance.

Activity-dependent neuroprotector homeobox protein level in Alzheimer's disease in Taiwanese - Corrected Proof

2012-05-07

Abstract: Alzheimer's disease (AD) is the most common cause of dementia of late life. The aim of this study was to utilize the proteomic approaches to establish serum protein patterns of AD. By using nano-high-performance liquid chromatography/electrospray ionization tandem mass spectrometry followed by peptide fragmentation pattern software to analyze proteome in human serum, the activity-dependent neuroprotector homeobox protein was found to exhibit significant differential expression compared with the control group and was confirmed by Western blotting and enzyme-linked immunosorbent assay. It may play an important role in slowing the progression of clinical symptoms of AD.

Thyroid lymphoma with mediastinum invasion after radiotherapy and chemotherapy: A case report and literature review - Corrected Proof

Joh-Jong Huang, Ming-Yii Huang — 2012-05-04

Abstract: We report herein the case of a 42-year-old woman having thyroid diffuse large B-cell lymphoma with initial presentation of right neck mass. After cytology and immunohistochemical profiles confirmed the diagnosis of a primary thyroid lymphoma, she received regional surgical excision, adjuvant radiotherapy, and systemic chemotherapy.