In vitro and ex vivo evaluation of a multi-epitope heparinase vaccine for various malignancies.

Previous studies have indicated that heparanase (Hpa) might represent a candidate universal tumor-associated antigen. However, vaccine therapy targeting only one cytotoxic T lymphocyte (CTL) epitope is suboptimal in preventing cancer. In the present study, we designed heparanase multi-epitope vaccines to increase the immune response to standard single heparanase epitopes. The results showed that multi-epitope vaccines Hpa525 + 277 + 405 + 16 and Hpa8 + 310 + 315 + 363 induced higher Hpa-specific lysis of various cancer cells from different tissues in a HLA-A2-restricted and heparanase-specific manner compared with the single epitope vaccines Hpa525, Hpa277, Hpa405, Hpa16, Hpa8, Hpa310, Hpa315 and Hpa363, both in vitro and ex vivo. Heparanase multi-epitope vaccines not only induced the heparanase-specific CTL to lyse tumor cells but also increased CTL secretion of interferon-γ. However, these heparanase-specific CTL did not lyse heparanase-expressing autologous lymphocytes and dendritic cells, which confirms the safety of these multi-epitope vaccines. Therefore, the present study provides theoretical evidence for the use of heparanase multi-epitope vaccines for clinical application.

Efficacy and safety of esomeprazole with flupentixol/melitracen in treating gastroesophageal reflux disease patients with emotional disorders.

BACKGROUND AND AIM: This study was designed to demonstrate the safety and efficacy of esomeprazole combined with flupentixol/melitracen for the treatment of gastroesophageal reflux disease (GERD) patients with emotional disorders. METHODS Two hundred eighty-nine GERD patients with emotional disorders were divided randomly into two groups: group 1 received esomeprazole only (monotherapy) and group 2 received esomeprazole and flupentixol/melitracen (combination therapy). The patients' GERD questionnaire (GerdQ) and hospital anxiety and depression (HAD) scores were obtained before and after treatment. Changes in the scores, rates of symptom remission, and adverse effects were compared between the two groups. RESULTS: After 2 weeks of treatment, the average decrease in GerdQ score in the combination group (4.04 ± 2.34) was significantly greater than that in the monotherapy group (3.34 ± 2.74; P < 0.05). Significant differences between the two groups were also found for changes in HAD anxiety scores (5.45 ± 2.41 vs 3.34 ± 2.43, P < 0.05), depression scores (5.47 ± 2.47 vs 3.00 ± 3.28, P < 0.05), and anxiety-depression scores (5.20 ± 2.71 vs 3.60 ± 2.56, P < 0.05). The remission of symptoms (eructation, abdominal pain, anorexia, and other accompanying symptoms) in the combination group was significantly better than that in the monotherapy group, and no significant difference in the incidence of adverse events was observed between the two groups. CONCLUSIONS: The combination therapy has better efficacy than the monotherapy in improving the symptoms of gastroesophageal reflux in patients with emotional disorders. In addition, this combination treatment is safe, with a low incidence of adverse events.

MiR-26a regulates cell cycle and anoikis of human esophageal adenocarcinoma cells through Rb1-E2F1 signaling pathway.

Resistance to anoikis, the subtype of apoptosis induced by lack of matrix adhesion, contributes to malignant transformation and development of metastasis. MicroRNAs play key regulatory roles in tumorigenesis and metastasis. In this study, we described that miR-26a, which is usually downregulated in tumor cells, is involved in the acquisition of anoikis-resistance of human esophageal adenocarcinoma (EA) cells. Results of qRT-PCR in clinical samples showed that downregulated miR-26a expression is related to tumorigenesis and metastasis of EA. In vitro experiments determined that miR-26a directly participates in the regulation of cell cycle and anoikis of human EA OE33 cells. Further, we identified that Rb1 is the direct functional target of miR-26a, and revealed that the reduction of miR-26a expression leads to increased Rb1 protein level and thus inhibits the function of E2F1, by which it influences the phenotypes of cell cycle and anoikis. The findings we reported here presented the evidence that miR-26a may be involved in regulation of anoikis-resistance of EA cells. Targeting miR-26a may provide a novel strategy to inhibit metastasis.

Antitumor effect of new multiple antigen peptide based on HLA-A0201-restricted CTL epitopes of human telomerase reverse transcriptase (hTERT).

The development of peptide vaccines aimed at enhancing immune responses against tumor cells is becoming a promising area of research. Human telomerase reverse transcriptase (hTERT) is an ideal universal target for novel immunotherapies against cancers. The aim of this work was to verify whether the multiple antigen peptides (MAP) based on HLA-A0201-restricted CTL epitopes of hTERT could trigger a better and more sustained CTL response and kill multiple types of hTERT-positive tumor cells in vitro and ex vivo. Dendritic cells (DC) pulsed with MAP based on HLA-A0201-restricted CTL epitopes of hTERT (hTERT-540, hTERT-865 and hTERT-572Y) were used to evaluate immune responses against various tumors and were compared to the immune responses resulting from the use of corresponding linear epitopes and a recombinant adenovirus-hTERT vector. A 4-h standard (51) Cr-release assay and an ELISPOT assay were used for both in vitro and ex vivo analyses. Results demonstrated that targeting hTERT with an adenovector was the most effective way to stimulate a CD8(+) T cell response. When compared with linear hTERT epitopes, MAP could trigger stronger hTERT-specific CTL responses against tumor cells expressing hTERT and HLA-A0201. In contrast, the activated CTL could neither kill the hTERT-negative tumor cells, such as U2OS cells, nor kill HLA-A0201 negative cells, such as HepG2 cells. We also found that these peptide-specific CTL could not kill autologous lymphocytes and DC with low telomerase activity. Our results indicate that MAP from hTERT can be exploited for cancer immunotherapy.

Noninvasive and real-time monitoring of the therapeutic response of tumors in vivo with an optimized hTERT promoter.

BACKGROUND: Telomerase is commonly recognized as an effective anticancer target. The human telomerase reverse transcriptase (hTERT), the rate-limiting component of telomerase, is expressed in most malignant tumors, but it is not found in most normal somatic cells. Here, we report a real-time and noninvasive method to monitor tumor response to a lentivirus-based hTERT-conditional suicidal gene therapy. METHODS: In this study, we constructed a lentivirus system in which an optimized hTERT promoter was used to drive the expression of the cytosine deaminase (CD) gene, one of the suicide genes, and a green fluorescent protein (GFP) reporter gene (pLenti-CD/GFP). The lentivirus was used to infect telomerase-positive or telomerase-negative cell lines. In vitro and in vivo experiments were conducted to analyze the dynamic processes of exogenous gene expression noninvasively in cell culture and living animals in real time via optical imaging. RESULTS: The lentivirus was able to express the CD gene and GFP in telomerase-positive tumor cells and significantly decrease cell proliferation after the use of prodrug 5-flucytosine. However, it could not express GFP and CD in telomerase-negative cell lines, nor could it induce any suicidal effect in those cells. The in vivo study showed that telomerase-positive tumors can be visualized after intratumor injection of the lentivirus in tumor-bearing nude mice via an optical imaging system. Significant tumor growth suppression was observed in telomerase-positive tumors. CONCLUSIONS: Collectively, this technology provides a valuable, noninvasive method to evaluate the real-time therapeutic response of tumors in vivo.

[Corrigendum] Downregulation of heparanase by RNA interference inhibits invasion and tumorigenesis of hepatocellular cancer cells in vitro and in vivo.

Following the publication of this article, an interested reader drew to the authors' attention that two images in Fig. 1B (the a and d panels) appeared to represent the same clone, albeit with different intensities and the panels were cropped differently. The authors were able to confirm that Figs. 1B(a) and B(d) were inadvertently selected from the same set of images but with different exposure times: Owing to an error in data handling, a wrong image was chosen during the grouping the figures. The corrected version of Fig. 1 is shown on the next page, featuring the correct image for Fig. 1B(d). The authors regret that this error was not picked up upon before the paper was sent to press, although the error did not affect the major conclusions reported in the paper. The authors thank the Editor of International Journal of Oncology for allowing them the opportunity to publish a Corrigendum. and regret any inconvenience caused to the readership. [the origional article was published on International Journal of Oncology 40: 1601­1609, 2012; DOI: 10.3892/ijo.2012.1338].

Cytotoxic T lymphocyte epitopes from human heparanase can elicit a potent anti-tumor immune response in mice.

Heparanase is expressed in almost all advanced tumors, and therefore it may serve as a potential target for tumor therapy. Our previous study has shown that heparanase can serve as a potential universal tumor-associated antigen (TAA) for the immunotherapy of advanced tumors. Further study demonstrated that the HLA-A*0201-restricted Cytotoxic T lymphocytes (CTL) epitopes Hpa525 (PAFSYSFFV), Hpa277 (KMLKSFLKA) and Hpa405 (WLSLLFKKL) from human heparanase could induce a potent anti-tumor immune response in vitro. The present study was designed to investigate whether the above peptides could induce immune responses in mice. Our results demonstrated that the effectors from heparanase peptide-immunized mice could effectively lyse various tumor cells that were heparanase positive and HLA-A*0201 matched. We also found that these peptide-specific CTLs did not lyse autologous lymphocytes that had low heparanase activity. Further study revealed that Hpa525, Hpa277, and Hpa405 peptides increased the frequency of IFN-gamma-producing T cells as compared to a negative peptide. These results suggest that Hpa525, Hpa277, and Hpa405 peptides are novel HLA-A*0201-restricted CTL epitopes capable of inducing heparanase-specific CTLs in mice. Because heparanase is expressed in most advanced malignant tumors, Hpa525, Hpa277, and Hpa405 peptide-based vaccines may be useful for the immunotherapy of patients with advanced tumors.

Induction of anti-tumour immunity by dendritic cells transduced with hTERT recombinant adenovirus in mice.

Dendritic cells (DCs) transfected with recombinant, replication-defective adenovirus (Ad) vectors encoding the human telomerase reverse transcriptase (hTERT) are potent inducers of cytotoxic T lymphocytes (CTLs) and anti-tumour immunity. However, previous studies have mostly been in vitro. In this study, we sought to determine whether DCs transfected with hTERT (DC/Ad-hTERT) could elicit a potent anti-tumour immunogenic response in vivo. We found that murine DCs transfected with recombinant adenovirus encoding the hTERT gene (DC/Ad-hTERT) induced hTERT-specific CTLs in vivo effectively, compared with Ad-LacZ-transduced DC (DC/Ad-LacZ) controls. These hTERT-specific CTLs lysed various tumour cell lines in an hTERT-specific and MHC-I molecule-restricted fashion. We also found that DC/Ad-hTERT could increase antigen-specific T-cell proliferation and augment the number of IFN-gamma secreting T-cells in mice. These data suggest that the DC/Ad-hTERT vaccine may induce anti-tumour immunity against tumour cells expressing hTERT in an MHC-I molecule-restricted fashion in vivo through the augmentation of the hTERT-specific CTL response. The DC/Ad-hTERT vaccine may thus be used as an efficient DC-based tumour vaccine in clinical applications.

hTERT promotes the invasion of telomerase-negative tumor cells in vitro.

Telomerase activation is a common feature of most types of human cancers. Although several studies have shown that activation of telomerase might participate in the progression of tumors, the mechanism by which telomerase activation causes the invasion and metastasis of tumors remains unclear. In this study, we transfected a vector containing the full-length cDNA of hTERT into a telomerase-negative osteosarcoma cell line U2OS (hTERT/U2OS). Vacant vector-transfected U2OS cells served as a control (EGFP/U2OS). We then compared the biological and vitodynamic changes in these transfected and untransfected U2OS cells. The hTERT protein was detected in hTERT/U2OS cells by Western blot analysis and immunochemistry assay. The telomere length in hTERT/U2OS cells was longer than that in EGFP/U2OS and untransfected U2OS cells. We also found using vacuum micropipette aspiration that hTERT transfection did not only promote the proliferation of hTERT-transfected U2OS cells but also increased the cellular adhesion capacity to the extracellular matrix. Transwell matrigel assay confirmed an increased invasion ability in hTERT/U2OS cells. These results strongly suggest that hTERT transfection promotes the invasion of telomerase-negative cells. Telomerase-mediated telomere maintenance enables these cells to achieve a fully malignant endpoint, including invasion and metastasis.

Acid and bile salt up-regulate BMP4 expression in human esophageal epithelium cells.

OBJECTIVE: Barrett's esophagus (BE) with an intestinal-type epithelium is thought to be a precancerous lesion of adenocarcinoma of the esophagus. The pathophysiology of Barrett's metaplasia is poorly understood. Previous studies suggest that differentiation of multipotent cells to columnar epithelium may be one of the possible mechanisms. Bone morphogenetic protein 4 (BMP4), a factor determining the fate of cells, is up-regulated in BE and esophagitis mucosa when compared with normal squamous or non-goblet cell-containing cardiac epithelium. The aim of this study was to demonstrate that BMP4 is a molecular mediator that links etiological agents of BE to the phenotypic changes in human esophagus epithelium cells (HEECs). MATERIAL AND METHODS: Primary cultured HEECs were used to investigate the effect of acid and bile salt on BMP4 expression and to examine the biological effects of BMP4 on HEECs. RESULTS: Acid and bile salt increased the expression of BMP4. In addition, recombinant human BMP4 induced villin expression in HEECs, as did chronic acid exposure, which can be effectively inhibited by Noggin, a specific antagonist of BMP4. Results from a Western blot assay suggest that BMP4 induces activation of smad1 and promotes protein expression of ID2 and CDX2. CONCLUSION: BMP4 may play an important role in the development of BE.

Down-regulated expression of cytochrome P450 (CYP) involved in the development and progression of esophageal adenocarcinoma.

BACKGROUND/AIMS: To analyze the differential expression genes (DEGs) between esophageal adenocarcinoma (EAC) and para-cancerous tissue (PCT) and explore the target genes related to the development and progression of EAC. METHODOLOGY: The total RNAs of matched EAC and para-cancerous tissue of EAC patients were isolated using one step Trizol method. Matched RNAs were qualified using 10 g/L agarose gel electrophoresis. cRNAs were synthesized, fluorescence labeled and purified after total RNAs were purified. The RNAs of EAC and PCT were hybridized with Agilent oligomicroarray (30,968 probes). The fluorescence intensity features were detected by Agilent scanner and quantified by feature extraction software. RESULTS: (1)The total RNA, reverse transcription product and fluorescence labeled cRNA were all of high quality; (2)There were 212 up-regulated genes and 126 down-regulated genes am- ong 2-fold DEGs, including 16 genes related to cytochrome P450 (CYP). CONCLUSIONS: Many EAC-assoeiated genes were screened by the high-throughput gene chip method. The development and progression of EAC is a complicated process involving multigenes and multiprocedures. The down-regulated expression of CYP related genes and gene polymorphism of CYP2 subfamily may be involved in the onset and progress of EAC.

Helicobacter pylori damages human gallbladder epithelial cells in vitro.

AIM: To study the mechanism by which Helicobacter pylori (H pylori) damages human gallbladder epithelial cells (HGBEC). METHODS: H pylori isolated from gallbladder were cultured in a liquid medium. Different concentration supernatants and sonicated extracts of H pylori cells were then added to HGBEC in a primary culture. The morphological changes in HGBEC as well as changes in the levels of alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and glutamyltransferase (GGT) were measured. RESULTS: According to the culture curve of HGBEC, it was convenient to study the changes in HGBEC by adding H pylori sonicated extracts and H pylori culture supernatants. Both H pylori sonicated extracts and H pylori culture supernatants had a significant influence on HGBEC morphology, i.e. HGBEC grew more slowly, their viability decreased and their detachment increased. Furthermore, HGBEC ruptured and died. The levels of ALP (33.84+/-6.00 vs 27.01+/-4.67, P<0.05), LDH (168.37+/-20.84 vs 55.51+/-17.17, P<0.01) and GGT (42.01+/-6.18 vs 25.34+/-4.33, P<0.01) significantly increased in the HGBEC culture supernatant in a time- and concentration-dependent. The damage to HGBEC in H pylori culture liquid was more significant than that in H pylori sonicated extracts. CONCLUSION: H pylori induces no obvious damage to HGBEC.

[Characteristics of cortical mapping in response to acid exposure in non-erosive reflux disease and erosive esophagitis using functional magnetic resonance imaging].

OBJECTIVE: To investigate the brain mechanisms for esophageal visceral hypersensitivity. METHODS: Thirty-one non-erosive reflux disease (NERD) patients, 21 in the group of NERD with esophageal hypersensitivity (NERD-H) and 10 in the group of NERD with normal esophageal sensation (NERD-N), 13 patients with erosive esophagitis (EE), and 12 healthy volunteers, all sex- and age-matched, underwent whole brain blood oxygenation level dependent (BOLD) fictional magnetic resonance imaging (fMRI) to record the cortical fMRI response to intraesophageal perfusion of normal saline or dilute hydrochloric acid. RESULTS: The main centers affected in the NERD-H patients included the secondary somatosensory cortex (SII), primary somatosensory cortex (S1), right prefrontal cortex (PFC), right orbitofrontal cortex (OFC), insular cortex, amygdala, striatum, motor cortex and its supplementary area, and cerebellum cortices, which form part of the matrix controlling emotional, autonomic modulatory responses to pain. The peak fMRI signal intensity and average maximum percent signal increase in the regions of interest (ROI) at above-mentioned brain areas of the NERD-H group were significantly stronger than those of the NERD-N and control groups (all P < 0.01). The peak image intensity of the anterior cingulate gyrus (ACC) of NERD-H group was 562 +/- 104, significantly lower than that of the control group (587 +/- 126, P < 0. 05), but significantly higher than that of the EE group (535 +/- 91, P < 0.05). The timeline of activation and deactivation events of particular ROI differentiate the four groups. The initial image latency and peak fMRI latency after hydrochloric acid perfusion of the NERD-H patients were 1.7 min +/- 0.9 min and peak 4. 5 min +/- 1.3 min respectively, both significantly shorter than those of the NERD-N group (4.0 min +/- 1.1 min and 6.8 min +/- 1.6 min respectively, both P < 0.01) and those of the control group (5. 4 min +/- 1.7 min and 7.2 min +/- 1.5 min respectively, both P < 0.01). The range of deactivation of SII and R-PFC of the NERD-H group were 26.5% +/- 5.4% and 20.3% +/- 3. 0% respectively, both significantly greater than those of the NERD-N group (8.2% +/- 2.2% and 16.4% +/- 3.6% respectively, both P < 0.05) and those of the EE group (11.9% +/- 4.8% and 11.7% +/- 3.1% respectively, both P < 0.01). The range of deactivation in ACC of the control group was 16.9% +/- 2.5%, significantly greater than those of the NERD-H and EE groups (11.8% +/- 2.8% and 6.4% +/- 1.0% respectively, both P < 0.05). CONCLUSION: The function of central nervous system to integrate and manage the convergence information becomes abnormal under the status of esophageal visceral hypersensitivity.

Effect of BN52021 on NFkappa-Bp65 expression in pancreatic tissues of rats with severe acute pancreatitis.

AIM: To investigate dynamic changes and significance of expression of NF-kappaBp65 in pancreatic tissues of rats with severe acute pancreatitis (SAP), as well as BN52021 effects. METHODS: Wistar male rats were randomly divided into negative control group (NC group, n=60), SAP-model group (SAP group, n=60), and BN52021-treated group (BN group, n=60), and each of the above groups was respectively divided into 6 subgroups at different time points after operation (1 h, 2 h, 3 h, 6 h, 12 h, and 24 h) (n=10). By RT-PCR and Western blot, NF-kappaBp65 mRNA and its protein expression in pancreatic tissues of rats were detected respectively. RESULTS: The expression of NF-kappaBp65 mRNA dynamically changed in both SAP groups and BN groups. The mRNA level was higher in SAP groups than NC groups at 2 h, 3 h, 12 h, and 24 h after operation (P<0.05), higher in BN groups than NC groups at all time points (P<0.05), and higher in BN groups than SAP group at 1 h (P<0.05). The NF-kappaBp65 protein level was higher in SAP groups than NC groups at 1 h, 3 h, and 6 h (P<0.01), and 2 h, 12 h, and 24 h (P<0.05), higher in BN groups than NC groups at all time points (P<0.05), and lower in BN groups than SAP groups at 1 h, 3 h, and 6 h (P<0.05). CONCLUSION: The expression of NF-kappaBp65 in pancreatic tissues is dynamically changed and the changes play an important role in pathogenesis of SAP. BN52021 exerts therapeutic effects through reducing the expression level of NF-kappaBp65 protein in the early stage of SAP.

H pylori exist in the gallbladder mucosa of patients with chronic cholecystitis.

AIM: To study whether H pylori locate in the gallbladder mucosa of patients with chronic cholecystitis. METHODS: Using Warthy-Starry (W-S) silver stain and immunohistochemistry stain with anti-H pylori antibodies, we screened paraffin specimens in 524 cases of cholecystitis. H pylori urease gene A (HPUA) and H pylori urease gene B (HPUB) were analyzed by polymerase chain reaction (PCR) in the fresh tissue specimens from 81 cases of cholecystitis. RESULTS: H pylori-like bacteria were found in 13.55% of the gallbladders of the cholecystitis patients using W-S stain. Meanwhile, bacteria positive for H pylori antibodies were also found in 7.1% of the gallbladders of patients with cholecystitis by immunohistochemistry. Of 81 gallbladders, 11 were positive for both HPUA and HPUB, 4 were positive for HPUA only and 7 were positive for HPUB only. CONCLUSION: H pylori exist in the gallbladders of patients with chronic cholecystitis.

H pylori are associated with chronic cholecystitis.

AIM: To study whether H pylori are associated with chronic cholecystitis. METHODS: The subjects were divided into three groups: H pylori-infected cholecystitis group, H pylori-negative cholecystitis group and control group. Pathologic changes of the gallbladder were observed by optic and electronic microscopes and the levels of interleukin-1, 6 and 8 (IL-1, 6 and 8) were detected by radioimmunoassay. RESULTS: Histological evidence of chronic cholecystitis including degeneration, necrosis, inflammatory cell infiltration, were found in the region where H pylori colonized. Levels of IL-1, 6 and 8 in gallbladder mucosa homogenates were significantly higher in H pylori-infected cholecystitis group than those in H pylori-negative cholecystitis group and control group. CONCLUSION: H pylori infection may be related to cholecystitis.

Pharmacological action and mechanisms of ginkgolide B.

OBJECTIVE: To review the recent research progress in pharmacological actions and mechanisms of ginkgolide B. Data sources Information included in this article was identified by searching of PUBMED (1987 - 2006) online resources using the key terms "ginkgolide B", "platelet activating factor", and "pharmacological". Study selection Mainly original milestone articles and critical reviews written by major pioneer investigators of the field were selected. RESULTS: The key issues related to the pharmacological actions and mechanisms of ginkgolide B were summarized. The ginkgolide B possesses a number of beneficial effects such as anti-inflammatory, anti-allergic, antioxidant, and neuroprotective effects. Meantime, their mechaniams were discussed. CONCLUSIONS: The Ginkgolide B is the most potent antagonist of platelet activating factor (PAF) and exhibits therapeutic action in a variety of diseases mainly by the PAF receptor.

[Mechanisms of human telomerase reverse transcriptase RNAi which increases hepatocellular carcinoma cell apoptosis induced by TRAIL].

OBJECTIVES: To investigate the mechanisms for human telomerase reverse transcriptase (hTERT) RNA interference (RNAi) in increasing hepatocellular carcinoma cell apoptosis induced by TNF-related apoptosis-inducing ligand (TRAIL). METHODS: Cell apoptosis was identified by flow cytometry analysis after annexin V/PI double staining. Expression of apoptosis-related proteins, procaspase-8, -9, -3, Bax, Bcl-2 and hTERT, were identified by Western blotting analysis; telomerase activity and telomere length were detected by telomeric repeat amplification protocol (TRAP) and telomere amount and length assay (TALA) methods. RESULTS: Hepatocellular carcinoma cell apoptosis induced by TRAIL were all significantly increased by hTERT RNAi (P less than 0.05). For example, apoptosis rates were enhanced from 5.53% (untransformed) to 10.35% (transformed) in HepG 2 cells and from 14.73% to 77.24% in SMMC 7721 cells after being treated by 100 ng/ml TRAIL for 24 h. Moreover, activation of procaspase-8, -9 and -3 in transformed cells after being treated by TRAIL were all significantly raised (P less than 0.05) in a dose-dependent manner. The expression of procaspase-8, -9 and Bcl-2 were effectively augmented (P less than 0.05), but expressions of Bax and hTERT were strikingly decreased (P less than 0.05). Meanwhile, telomerase activity was apparently suppressed and telomere length was markedly shortened (P less than 0.05). There were no remarkable differences in these effects between control cells and the untransformed cells (P more than 0.05). CONCLUSION: Enhanced cell apoptosis induced by TRAIL through hTERT RNAi may be related to up-regulation of procaspase-8 and -9 expressions. However the down-regulation of hTERT expression, reduced telomerase activity and shortened telomere length may not be related to expressions of Bcl-2 and Bax.

Optimized high-dose amoxicillin-proton-pump inhibitor dual therapies fail to achieve high cure rates in China.

Background\Aim: Quadruple daily administration of proton-pump inhibitor (PPI) therapy achieves potent acid inhibition, and combined with amoxicillin, with its pharmacodynamic and pharmacokinetic characteristics, may be efficient for Helicobacter pylori eradication. We compared the efficacy of two optimized high-dose dual therapies with a bismuth-containing quadruple regimen for treating H. pylori infection. Rabeprazole dosages for H. pylori eradication were also evaluated. PATIENTS AND METHODS: Treatment-naive and H. pylori-positive subjects were recruited and randomly apportioned to three treatment groups: Group A (n = 87), rabeprazole 10 mg plus amoxicillin 750 mg (4 times/day for 14 days); Group B (n = 87), rabeprazole 20 mg plus amoxicillin 750 mg (4 times/day for 14 days); and Group C (n = 89), bismuth-containing quadruple regimen consisting of rabeprazole 20 mg, bismuth 220 mg, amoxicillin 1000 mg, and clarithromycin 500 mg (2 times/day for 14 days). Four weeks after treatment discontinuation, patients were examined for H. pylori infection by 13C-urea breath test. The rates of adverse effects, compliance, and eradication were evaluated. RESULTS: Eradication rates in groups A, B, and C were 78.1, 81.6, and 84.3%, respectively, based on intention-to-treat analysis, or 79.1, 83.5, and 86.2%, according to per-protocol analysis. Rates of adverse events and compliance of the three groups were similar. CONCLUSION: For treating H. pylori infection, optimized high-dose amoxicillin-PPI dual therapies failed to achieve high cure rates in China and held no advantage over a bismuth-containing quadruple regimen.

Dynamic functional and ultrastructural changes of gastric parietal cells induced by water immersion-restraint stress in rats.

AIM: To investigate the dynamic functional and ultrastructural changes of gastric parietal cells induced by water immersion-restraint stress (WRS) in rats. METHODS: WRS model of Sprague-Dawley (SD) rats was established. Fifty-six male SD rats were randomly divided into control group, stress group and post-stress group. The stress group was divided into 1, 2 and 4 h stress subgroups. The post-stress group was divided into 24, 48 and 72 h subgroups. The pH value of gastric juice, ulcer index (UI) of gastric mucosa and H(+), K(+)-ATPase activity of gastric parietal cells were measured. Ultrastructural change of parietal cells was observed under transmission electron microscope (TEM). RESULTS: The pH value of gastric juice decreased time-dependently in stress group and increased in post-stress group. The H(+), K(+)-ATPase activity of gastric parietal cells and the UI of gastric mucosa increased time-dependently in stress group and decreased in post-stress group. Compared to control group, the pH value decreased remarkably (P = 0.0001), the UI and H(+), K(+)-ATPase activity increased significantly (P = 0.0001, P = 0.0174) in 4 h stress subgroup. UI was positively related with stress time (r = 0.9876, P < 0.01) but negatively with pH value (r = -0.8724, P < 0.05). The parietal cells became active in stress group, especially in 4 h stress subgroup, in which plenty of intracellular canalicular and mitochondria were observed under TEM. In post-stress group, the parietal cells recovered to resting state. CONCLUSION: The acid secretion of parietal cells is consistent with their ultrastructural changes during the development and healing of stress ulcer induced by WRS and the degree of gastric mucosal lesions, suggesting gastric acid play an important role in the development of stress ulcer and is closely related with the recovery of gastric mucosal lesions induced by WRS.