Browsing by Author "Wu, Chun-Ying"

Context Tumor recurrence is a major issue for patients with hepatocellular carcinoma (HCC) following curative liver resection. Objective To investigate the association between nucleoside analogue use and risk of tumor recurrence in patients with hepatitis B virus (HBV)-related HCC after curative surgery. Design, Setting, and Participants A nationwide cohort study between October 2003 and September 2010. Data from the Taiwan National Health Insurance Research Database. Among 100 938 newly diagnosed HCC patients, we identified 4569 HBV-related HCC patients who received curative liver resection for HCC between October 2003 and September 2010. Main Outcome Measures The risk of first tumor recurrence was compared between patients not taking nucleoside analogues (untreated cohort, n=4051) and patients taking nucleoside analogues (treated cohort, n=518). Cumulative incidences and hazard ratios (HRs) were calculated after adjusting for competing mortality. Results The treated cohort had a higher prevalence of liver cirrhosis when compared with the untreated cohort (48.6% vs 38.7%; P<.001), but lower risk of HCC recurrence (n=106 [20.5%] vs n=1765 [43.6%]; P<.001), and lower overall death (n=55 [10.6%] vs n=1145 [28.3%]; P<.001). After adjusting for competing mortality, the treated cohort had a significantly lower 6-year HCC recurrence rate (45.6%; 95% CI, 36.5%-54.6% vs untreated, 54.6%; 95% CI, 52.5%-56.6%; P<.001). Six-year overall mortalities for treated cohorts were 29.0% (95% CI, 20.0%-38.0%) and for untreated 42.4% (95% CI, 40.0%-44.7%; P<.001). On modified Cox regression analysis, nucleoside analogue use (HR, 0.67; 95% CI, 0.55-0.81; P<.001), statin use (HR, 0.68; 95% CI, 0.53-0.87; P=.002), and nonsteroidal anti-inflammatory drugs or aspirin use (HR, 0.80; 95% CI, 0.73-0.88; P<.001) were independently associated with a reduced risk of HCC recurrence. Multivariable stratified analyses verified the association in all subgroups of patients, including those who were noncirrhotic (HR, 0.56; 95% CI, 0.42-0.76) and diabetic (HR, 0.52; 95% CI, 0.31-0.89). Conclusion Nucleoside analogue use was associated with a lower risk of HCC recurrence among patients with HBV-related HCC after liver resection. JAMA. 2012; 308(18): 1906-1913 Published online November 12, 2012. doi: 10.1001/2012.jama.11975 www.jama.com

It is unclear whether dysbiosis in hepatitis C virus (HCV) infected patients results from the viral infection per se or develops as a result of hepatic dysfunction. We aimed to characterize compositions in gut microbiome before and shortly after HCV clearance. In this prospective cohort study, adult patients with confirmed HCV viremia were screened before receiving direct antiviral agents. Those with recent exposure to antibiotics or probiotics (within one month), prior abdominal surgery, or any malignancy were ineligible. Stool was collected before antiviral therapy started and at 12 weeks after the treatment completed. From the extracted bacterial DNA, 16 s rRNA gene was amplified and sequenced. Each patient was matched 1:2 in age and sex with uninfected controls. A total of 126 individuals were enrolled into analysis. The gut microbiome was significantly different between HCV-infected patients (n = 42), with or without cirrhosis, and their age-and sex-matched controls (n = 84) from the levels of phylum to amplicon sequence variant (all p values < 0.01 by principal coordinates analysis). All patients achieved viral eradication and exhibited no significant changes in the overall composition of gut microbiome following viral eradication (all p values > 0.5), also without significant difference in alpha diversity (all p values > 0.5). For the purpose of exploration, we also reported bacteria found differently abundant before and after HCV eradication, including Coriobacteriaceae, Peptostreptococcaceae, Staphylococcaceae, Morganellaceae, Pasteurellaceae, Succinivibrionaceae, and Moraxellaceae. Gut microbiota is altered in HCV-infected patients as compared with uninfected controls, but the overall microbial compositions do not significantly change shortly after HCV eradication.

Emerging laboratory and animal studies suggest that aspirin may prevent non-alcoholic fatty liver disease (NAFLD)-related hepatocellular carcinoma (HCC), however clinical evidence remains lacking.

Background & Aims: We aimed to compare the efficacy of genotypic resistance–guided therapy vs empirical therapy for eradication of refractory Helicobacter pylori infection in randomized controlled trials. Methods: We performed 2 multicenter, open-label trials of patients with H pylori infection (20 years or older) failed by 2 or more previous treatment regimens, from October 2012 through September 2017 in Taiwan. The patients were randomly assigned to groups given genotypic resistance–guided therapy for 14 days (n = 21 in trial 1, n = 205 in trial 2) or empirical therapy according to medication history for 14 days (n = 20 in trial 1, n = 205 in trial 2). Patients received sequential therapy containing esomeprazole and amoxicillin for the first 7 days, followed by esomeprazole and metronidazole, with levofloxacin, clarithromycin, or tetracycline (doxycycline in trial 1, tetracycline in trial 2) for another 7 days (all given twice daily) based on genotype markers of resistance determined from gastric biopsy specimens (group A) or empirical therapy according to medication history. Resistance-associated mutations in 23S ribosomal RNA or gyrase A were identified by polymerase chain reaction with direct sequencing. Eradication status was determined by 13C-urea breath test. The primary outcome was eradication rate. Results: H pylori infection was eradicated in 17 of 21 (81%) patients receiving genotype resistance–guided therapy and 12 of 20 (60%) patients receiving empirical therapy (P =.181) in trial 1. This trial was terminated ahead of schedule due to the low rate of eradication in patients given doxycycline sequential therapy (15 of 26 [57.7%]). In trial 2, H pylori infection was eradicated in 160 of 205 (78%) patients receiving genotype resistance–guided therapy and 148 of 205 (72.2%) patients receiving empirical therapy (P =.170), according to intent to treat analysis. The frequencies of adverse effects and compliance did not differ significantly between groups. Conclusions: Properly designed empirical therapy, based on medication history, is an acceptable alternative to genotypic resistance–guided therapy for eradication of refractory H pylori infection after consideration of accessibility, cost, and patient preference. ClinicalTrials.gov ID: NCT01725906. ? 2018 AGA Institute

A new approach by investigating the intra-tumoral microbiome raised great interest because they may influence the host immune response and natural history of the disease. However, previous studies on the intra-tumoral microbiome of pancreatic ductal adenocarcinoma (PDAC) were mostly based on examining the formalin-fixed paraffin-embedded tumor specimens. This study aims to investigate the feasibility of using endoscopic ultrasound-guided fine-needle biopsy (EUS-FNB) as a complementary procedure of surgical biopsy to obtain adequate fresh pancreatic cancer tissue for intra-tumoral microbial research. This was a prospective pilot study performed at a single tertiary referral center. We obtained pancreatic cancer tissue by EUS-FNB and surgical biopsy, respectively. We amplified the V3-V4 hyper-variable region of bacterial 16S ribosomal ribonucleic acid (rRNA) genes, constructed a pair-end library, and performed high-throughput sequencing. From August 2020 to November 2020, nine eligible patients with PDAC were enrolled in this study. The intra-tumoral microbiome profile was successfully generated from the PDAC cancer tissue obtained by EUS-FNB as well as by surgical biopsy. There was no significant difference in intra-tumoral alpha-diversity or bacterial taxonomic composition between tissues obtained by EUS-FNB and by surgical biopsy. EUS-FNB can collect sufficient fresh cancer tissue for microbiome analyses without complication. The intra-tumoral microbiome profile in tissues obtained by EUS-FNB had similar alpha-diversity and taxonomic profiles with those obtained by surgical biopsy. It implicated, except for surgical biopsy, EUS-FNB can be another valid and valuable tool for studying intra-tumoral microbiome in patients with resectable and unresectable PDAC.

Hepatocellular carcinogenesis of hepatitis B virus (HBV) infection may arise from integration of viral DNA into the host genome. We aimed to gauge the effect of viral inhibition on transcriptionally active HBV-host integration events and explore the correlation of viral integrations with host gene dysregulation.

Background: Esophageal cancer has been associated with oral bisphosphonate use, but current data are conflicting and devoid of Asian studies where esophageal squamous carcinoma prevails. Methods: We assessed the association between dose intensity, stratified by use duration (observation period) and exposure frequency, of oral bisphosphonates and the risk of esophageal cancer using 16,204 esophageal cancer cases and 64,816 malignancy-free controls identified from the population-based National Health Insurance Research Database of Taiwan from 1997 to 2008. Results: Neither duration nor frequency of bisphosphonate exposures was positively correlated to esophageal cancer risk. The ORs for rare users of 1-, 3-, 5-year observation periods were 3.86, 2.58, and 2.27, respectively (P < 0.001). Similar trend of descending ORs was also observed for rare-, frequent-, and regular users of 1-year observation period (ORs = 3.86, 1.93, and 0.95, respectively). Conclusion: Our data suggest that bisphosphonates are not likely risk factors for esophageal cancer in Taiwan. Impact: The study shows no evidence of an association between bisphosphonate use and esophageal cancer risk from Asian perspective. ?2012 AACR.

Levofloxacin-based therapy or bismuth-based quadruple therapy are the recommended second-line regimens for Helicobacter pylori eradication after failure of clarithromycin-based therapy. However, resistance to levofloxacin has increased in the past decade. Furthermore, little is known about the long-term effects of H pylori eradication on the antibiotic resistome. In this study, we compared these second-line eradication therapies for efficacy, tolerability, and short-term and long-term effects on the gut microbiota, antibiotic resistome, and metabolic parameters.

Finite nucleos(t)ide analogue (NUC) therapy has been proposed as an alternative treatment strategy for chronic hepatitis B (CHB).

INTRODUCTION Gastric cancer remains a leading cause of cancer mortality, despite a worldwide decline in incidence. In Asian countries, gastric cancer is one of the most prevalent tumor and the leading cause of cancer death (Gordon D.Luk, 2005). In the Western world, more than 80% of gastric cancer patients have advanced cancer on diagnosis with poor prognosis (Roukos, 2000). Complete resection of the tumor and adjacent lymph nodes is the only proven, effective curative treatment (Kim, 1999). Unfortunately, the accuracy of current preoperative staging is limited, particularly with regard to depth of invasion, lymph node involvement and distant metastasis4. Developing new biomarkers to identify the subgroup of gastric cancer patients with invasive phenotypes will be helpful for avoiding inappropriate attempts at curative surgery. Matrix metalloproteinases (MMPs) are a family of zinc-dependent enzymes that breakdown the extracellular matrix (ECM) (Nagase and Woessner, Jr., 1999b). MMPs not only play important roles in physiologic ECM remodeling, but are also involved in pathological conditions, including tumor progression, invasion and metastasis (la-aho and Kahari, 2005;Egeblad and Werb, 2002b;Stamenkovic, 2003). Since tissue remodeling is often reflected in body fluids, measurements of MMPs in blood or urine have been suggested as useful tools for characterizing processes that occur in tissue (Zucker, et al, 1999). Among the MMPs family, MMP-9 (also known as 92-kDa gelatinase) is a promising new non-invasive marker (Zucker, et al, 1999). Elevated levels of serum or plasma MMP-9 have been found in a variety of malignant tumors, such as breast cancer, colon cancer, lung cancer, head and neck squamous cell carcinoma, hepatocellular carcinoma and gastric cancer (Hayasaka, et al, 1996; Hoikkala, et al, 2005; Ruokalainen, et al, 2005; Torii, et al, 1997; Endo et al, 1997; Shen et al, 2000; (Zucker et al., 1993). Although the usefulness of MMP-9 as a tumor marker has been established, several studies measuring MMP-9 in the peripheral blood of cancer patients, using serum or plasma samples, have produced controversial results (Torii et al, 1997; Kirman et al, 2006; Decock et al, 2005). Differences in enrolled populations and study designs most likely contribute to the discrepancies. However, blood sampling and processing may also influence the concentration of MMP-9. Several reports have highlighted the influence of blood specimen collection methods on MMP-9 concentrations (Jung et al, 205; Mannello et al, 2003). MMP-9 concentration has been found to be three-fold higher in serum than in heparin plasma (Jung et al, 1998). Platelet activation or neutrophil mobilization during clotting could produce such results (Makowski et al, 2001). Although measurements of MMP-9 in blood have been suggested to be performed in heparin plasma, rather than in serum (Jung et al, 2001), recent studies have used serum MMP-9 to investigate the correlation between MMP-9 and tumor progression. It is important to investigate how the differences between plasma and serum samples influence the diagnostic and prognostic performances of MMP-9. In the present study, we first compared the effectiveness of plasma and serum MMP-9 levels as tumor markers in gastric cancer. Then we examined whether plasma and serum MMP-9 levels correlate well with gastric cancer invasive phenotypes and survival. MMPs can be regulated by osteopontin (OPN) through OPN-uPA-MMPs pathway (Rangaswami, et al, 2006). The role of OPN in tumorigenesis can be explained by the multiple functions of OPN in cells (Rittling and Chambers, 2004). Several mechanisms have been proposed through studies using cultured cells. First, it is recognized that OPN has adhesive activity because its receptors all mediate cell adhesion. Second, the ability of cells to migrate may be directly tied to their tumorigenicity and OPN promotes the migration of diverse cells, including monocytes, macrophages and tumor cells, along OPN gradients (Denhardt et al, 2001). In addition, OPN-deficient cells are reported to be hypomotile (Zhu et al., 2004). Third, some experiments suggest that OPN inhibits apoptosis and stimulates survival and growth of cells with inducible OPN (Wu et al., 2000), or with the addition of OPN to cell culture medium (Chang et al., 2003), via an interaction with its receptor CD44 (Lin et al., 2000). Fourth, several studies have suggested that OPN increases tumor invasiveness by inducing proteinase, particularly uPA and MMPs, via complex signaling pathways, such as AP-1 activation, PI3-kiase/Akt-dependent or NIK-dependent NF-kB activation27-31. In the present study, we used real-time RT-PCR to demonstrate that OPN mRNA expression is significantly higher in gastric cancer tissues when compared with surrounding non-tumor tissues. This observation is compatible with a previous report using cDNA microarray method in which OPN is over-expressed in gastric cancer tissues5.Recent studies have consistently reported that OPN mRNA and protein expression in cancer tissues are closely related to invasion and metastasis of gastric cancer (Sun et al., 2005). However, the application of plasma OPN level as a biomarker for gastric cancer has not been investigated. MMP-2, also know as gelatinase A or 72 kDa collagenase IV, is a member of the MMP family which degrades gelatine and type IV collagen (Yu, et al. 2002). In contrast to other MMPs, MMP-2 is broadly, often constitutively, expressed by a large number of cell types and overexpressed in a wide variety of human cancers, including gastric, lung, prostate, ovarian and bladder cancers (Murray, et al., 1998; Miao, et al., 2003; Zhang, et al., 2005b; Zhou, et al., 2004; Upadhyay, et al, 1999; Vasala, et al., 2003; Davidson, et al., 1999b; Brown, et al, 1993). Human MMP-2 promoter has been shown to contain several cis-acting regulatory elements. Among them, a -1306 C→T transition interrupts Sp1 binding site and consequently diminishes promoter activity (Price, et al, 2001). Transient transfection experiments have shown that MMP-2 expression is ~1.4-2 fold higher with the C allele than with the T allele (Price, et al, 2001). The importance of Sp-1 binding activity in MMP-2 expression has also been reported in other MMP-2 promoter deletion or site-directed mutagenesis studies(Qin et al., 1999;Pan and Hung, 2002). These results suggest that patients with MMP-2 -1306 C/C genotype have higher MMP-2 expression than patients with C/T or T/T genotype. Recently, Miao and colleagues reported that -1306C/T is associated with gastric cardia adenocarcinoma risk (Miao, et al, 2003). Subjects with the C/C genotype had greater than three-fold risk for developing gastric cardia adenocarcinoma when compared with those with the variant C/T or T/T genotype (Miao, et al, 2003). The activity of MMP-2 is not only regulated by transcriptional regulation, but also by tissue inhibitors of metalloproteinases (TIMPs), which can form complexes either with latent or activated MMPs (Gomez et al., 1997a;Kahari and Saarialho-Kere, 1999a). Among the TIMP family, TIMP-2 is particularly interesting due to its dual functions of regulating MMP-2 activity (Howard et al., 1991b;Wang et al., 2000b) and its controversial effects on tumour progression(Egeblad andWerb, 2002a). TIMP-2 has been reported to be greater than 10 fold more effective than TIMP-1 in the inhibition of MMP-2 activation(Howard et al., 1991c). On the other hand, TIMP-2 has been found to be required for efficient activation of pro-MMP-2 in vivo (Wang et al., 2000a). ATIMP-2 promoter polymorphism (-418 G →C) has been identified, which is also located in the consensus sequence for the Sp-1 binding site (De Clerck et al., 1994a). Although the functional significance of this polymorphism is still unknown, it is reasonable to postulate that it interrupts the Sp-1 binding site and decreases TIMP-2 gene transcription, leading to MMP-2 and TIMP-2 imbalance (Hirano et al., 2001a). MMP-2 promoter polymorphism has been found to be associated with susceptibility to gastric cancer (Miao et al.,2003). However, there have been no studies conducted to elucidate the associations between MMP-2 polymorphism and gastric cancer invasive phenotype and survival. If MMP-2 polymorphism influences susceptibility to gastric cancer, it may also affect tumour progression and patient survival. As for TIMP-2 polymorphism, no studies have been conducted on gastric cancer patients. In the present study, we hypothesized that in gastric cancer, MMP-2 and TIMP-2 polymorphisms not only correlate well with susceptibility, but also with invasive phenotype and survival. A hospital-based case-control study was conducted to access this hypothesis. Urokinase type plasminogen activator (uPA), a member of the plasminogen activator (PA) family, converts plasminogen into plasmin, which can activate some prometalloproteinases and degrade the extracellular matrix (ECM) (Saksela and Rifkin, 1988). uPA is produced in both normal and malignant cells and plays important roles not only in tissue remodeling of normal cells, but also in degradation of ECM and destruction of the basement membrane of malignant cells (Dano et al., 1985). Involvement of uPA in diverse physiologic and pathologic processes, including inflammation (Gyetko et al., 1996), fibrinolysis (Myohanen and Vaheri, 2004), tumor growth stimulation (Blasi, 1993), invasion (Nekarda et al., 1994b), angiogenesis (Bacharach et al., 1992) and metastasis (Crowley et al., 1993), has been reported in recent years. In vitro studies have demonstrated that uPA activity inhibition results in the suppression of tumor progression and reduction of metastasis (Holst-Hansen et al., 1996). Clinically, poorer outcomes have been correlated with higher uPA expression in many types of tumors, including gastric cancer (Nekarda et al., 1994a;Yonemura et al., 1995;Heiss et al., 1995). Elevated uPA levels in gastric cancer tissue have been found to be associated with lymph node metastasis, venous invasion, serosal involvement and poor prognosis (Nekarda et al., 1994a;Heiss et al., 1995;Yonemura et al., 1995). Plasma uPA levels tend to be significantly increased in gastric cancer patients (Herszenyi et al., 2000). The roles of uPA expression in tumor occurrence, invasion and prognosis have been well established. However, how uPA genetic polymorphisms influence the occurrence and outcomes of tumors has not been widely investigated (Przybylowska et al., 2002). Two polymorphisms of the uPA gene have been described. Yoshimoto et al. reported a C→T transition in the nucleotide sequence of exon 6 encoding the kringle domain. The C →T transition results in Pro (CCG) to Leu (CTG) replacement at amino-acid position 121, which may alter the whole tertiary structure of uPA and be directly or indirectly involved in the activity of uPA (Yoshimoto et al., 1996). Conne et al. reported a T→C substitution in intron 7, located 7 bp upstream of the splicing acceptor site, which may be involved in the nuclear mRNA splicing process (Conne et al., 1997). In the present study, we investigated whether uPA exon 6 C/T and intron 7 T/C polymorphisms correlate well with gastric cancer susceptibility, invasion and survival.