Human Embryonic Stem Cells and Induced Pluripotent Stem Cells: The Promising Tools for Insulin-Producing Cell Generation

Diabetes mellitus, a disease with abnormally high level of blood glucose, can cause a wide range of chronic complications that affect almost every parts of the body. The major goal of diabetes treatments is to control elevated blood glucose without causing abnormally low levels of blood glucose. Despite islet transplantation provided endogenous insulin secretion in individuals with diabetes, the scarcity of cadaveric donors for pancreatic β-cell still remains a major obstacle. In this regard, the needs for an unlimited supply for cell replacement strategy have led to explore the way of generating insulin-producing cells to use in the disease treatment. Human embryonic stem cells (hESCs) offer a source to produce the desired kind of cell. Currently, several researchers achieved insulin-producing cells from hESCs using a multistep differentiation protocols, growth factors, and/or chemical compounds. In this review, we summarized the hESCs derivation, culture methods, and characteristics of hESCs. We also emphasized on the current methods for direct differentiation of hESCs into embryoid bodies (EBs) and toward insulin-producing cells, characterization of these insulin-producing cells, and the limitation of hESCs. Since the discovery of induced pluripotent stem cells (iPSCs), which have similar properties to hESCs but less ethical issues than hESCs, can be created directly from somatic cells that hold great promise as the therapeutic source for developing cell-based therapy. Herein, the methods to produce iPSC-derived insulin-producing cells are also discussed. Moreover, the encapsulation technology which is a powerful tool for accelerate hESCs and iPSCs applications in medicine which provide a new avenue for diabetes treatment in the future is also included in this review. Understanding the basic knowledge of hESCs and iPSCs, their differentiation capability toward insulin-producing cells will stimulate more therapeutic value of hESCs and iPSCs for diabetic treatments, drug screening, and regenerative medicine

WT1 gene expression as a prognostic marker in advanced serous epithelial ovarian carcinoma: an immunohistochemical study

BACKGROUND: WT1 is a tumor suppressor gene responsible for Wilms' tumor. WT1 reactivity is limited to ovarian serous carcinomas. Recent studies have shown that WT1 plays an important role in the progression of disease and indicates a poorer prognosis of human malignancies such as acute myeloid leukemia and breast cancer. The aims of this study were to determine the survival and recurrence-free survival of women with advanced serous epithelial ovarian carcinoma in relation to WT1 gene expression. METHODS: The study accrued women over an 18-year period, from 1987–2004. During the study period, 163 patients were diagnosed with advanced serous epithelial ovarian carcinoma and had undergone complete post-operative chemotherapy, but the final study group comprised 99 patients. The records of these women were reviewed and the paraffin-embedded tissue of these women stained with WT1 immunostaining. Survival analysis was performed using Kaplan-Meier and Cox regression methods. RESULTS: Fifty patients showed WT1 staining and forty-nine did not. Five-year survival of non-staining and staining groups were 39.4% and 10.7% (p < 0.00005); five-year recurrence-free survival of these groups were 29.8% and ≤ 7.5% (p < 0.00005), respectively. For survival the HR of WT1 staining, adjusted for residual tumor and chemotherapy response, was 1.98 (95% CI 1.28–3.79), and for recurrence-free survival the HR was 3.36 (95% CI 1.60–7.03). The HR for recurrence-free survival was not confounded by any other variables. CONCLUSION: This study suggests that expression of WT1 gene may be indicative of an unfavorable prognosis in patients with advanced serous epithelial ovarian carcinoma

Establishment of Insulin-Producing Cells From Human Embryonic Stem Cells Underhypoxic Condition for Cell Based Therapy

Diabetes mellitus (DM) is a group of diseases characterized by abnormally high levels of glucose in the blood stream. In developing a potential therapy for diabetic patients, pancreatic cells transplantation has drawn great attention. However, the hinder of cell transplantation for diabetes treatment is insufficient sources of insulin-producing cells. Therefore, new cell based therapy need to be developed. In this regard, human embryonic stem cells (hESCs) may serve as good candidates for this based on their capability of differentiation into various cell types. In this study, we designed a new differentiation protocol that can generate hESC-derived insulin-producing cells (hES-DIPCs) in a hypoxic condition. We also emphasized on the induction of definitive endoderm during embryoid bodies (EBs) formation. After induction of hESCs differentiation into insulin-producing cells (IPCs), the cells obtained from the cultures exhibited pancreas-related genes such as Pdx1, Ngn3, Nkx6.1, GLUT2, and insulin. These cells also showed positive for DTZ-stained cellular clusters and contained ability of insulin secretion in a glucose-dependent manner. After achievement to generated functional hES-DIPCs in vitro, some of the hES-DIPCs were then encapsulated named encapsulated hES-DIPCs. The data showed that the encapsulated cells could possess the function of insulin secretion in a time-dependent manner. The hES-DIPCs and encapsulated hES-DIPCs were then separately transplanted into STZ-induced diabetic mice. The findings showed the significant blood glucose levels regulation capacity and declination of IL-1β concentration in all transplanted mice. These results indicated that both hES-DIPCs and encapsulated hES-DIPCs contained the ability to sustain hyperglycemia condition as well as decrease inflammatory cytokine level in vivo. The findings of this study may apply for generation of a large number of hES-DIPCs in vitro. In addition, the implication of this work is therapeutic value in type I diabetes treatment in the future. The application for type II diabetes treatment remain to be investigated

Down-Regulation of C1GALT1 Enhances the Progression of Cholangiocarcinoma through Activation of AKT/ERK Signaling Pathways

Alteration of mucin-type O-glycosylation is implicated in tumor progression and metastasis of cholangiocarcinoma (CCA). Core 1 β1-3 Galactosyltransferase (C1GALT1) is a primary enzyme that regulates the elongation of core 1-derived mucin-type O-glycans. Dysregulation of C1GALT1 has been documented in multiple cancers and is associated with aberrant core 1 O-glycosylation and cancer aggressiveness; however, the expression of C1GALT1 and its role in CCA progression remains unknown. Our study demonstrated that C1GALT1 was downregulated in CCA tissues at both the mRNA and protein levels. The biological function of C1GALT1 using siRNA demonstrated that suppression of C1GALT1 in the CCA cell lines (KKU-055 and KKU-100) increased CCA progression, evidenced by: (i) Induction of CCA cell proliferation and 5-fluorouracil resistance in a dose-dependent manner; (ii) up-regulation of growth-related genes, ABC transporter genes, and anti-apoptotic proteins; and (iii) an increase in the activation/phosphorylation of AKT and ERK in silencing C1GALT1 cells. We demonstrated that silencing C1GALT1 in CCA cell lines was associated with immature core 1 O-glycosylation, demonstrated by high expression of VVL-binding glycans and down-regulation of other main O-linked glycosyltransferases β1,3-N-acetylglucosaminyltransferase 6 (B3GNT6) and ST6 N-Acetylgalactosaminide Alpha-2,6-Sialyltransferase 1 (ST6GALNAC1) in C1GALT1 knockdown. Our findings demonstrate that down-regulation of C1GALT1 in CCA increases the expression of immature core 1 O-glycan, enhancing CCA progression, including growth and 5-fluorouracil resistance via the activation of the AKT/ERK signaling pathway

Anti-tumour activities of fucoidan from the aquatic plant Utricularia aurea lour

Fucoidan, a sulfated polysaccharide with several biological activities, is usually isolated from marine seaweeds or from echinoderms. Here, we report on the anti-tumour activity of fucoidan isolated from the aquatic plant Utricularia aurea Lour (Lentibulariaceae). A crude extract (CE) prepared by incubating U.aurea with hot water at 95ºC for 12 hr was partially purified by Sephadex G-50, eluting with a 50mM sodium acetate buffer, at pH 5.0, containing 0.5M NaCl. Partially purified fucoidan (PPF) had a 3- fold increase in fucose content when compared with the CE and a molecular weigÄt of 11.6 kDa as determined by Sephadex G-200. Chemical analysis showed that CE consisted of 62.5% glucuronic acid, 5.0% fucose, 1.7% sulfate and 12.0% proteins while PPF consisted of 65.0% glucuronic acid, 15.3% fucose, 2.1 % sulfate and 8.3% proteins.The anti-tumour activity of the CE and PPF was determined by the MTT test. The CE at 125 μg/mL fucoidan and PPF at 250 μg/mL inhibited the growth of KB cells (a nasopharynx tumour cell line), but did not inhibit that of normal fibroblast cells. The inhibition was postulated to occur via apoptosis as significantlymore apoptotic cells were found after treatment than in the untreated KB cells (P<0.05) by the TUNNEL (TdT-mediated dUTP Nick-End Labelling) assay

Correlation between Gastric Mucosal Morphologic Patterns and Histopathological Severity of Helicobacter pylori Associated Gastritis Using Conventional Narrow Band Imaging Gastroscopy

Background and Aim. Identifying specific gastric mucosal morphologic patterns useful for detecting Helicobacter pylori associated gastritis and correlation with histopathological severity. Methods. The endoscopists classified the C-NBI gastroscopic findings into 5 gastric mucosal morphologic patterns as follows: type 1: regular arrangement of collecting venules, type 2: cone-shaped gastric pits, type 3: rod-shaped gastric pits with prominent sulci, type 4: ground glass-like morphology, and type 5: dark brown patches with bluish margin and irregular border. Biopsies of all of the cases were then evaluated by 5 pathologists for definitive Helicobacter pylori diagnosis. Result. Type 1 and type 2 patterns were statistically significant in predicting Helicobacter pylori negative status (58/60, P<0.01). Type 3, type 4, and type 5 patterns were statistically significant in predicting Helicobacter pylori positive status (132/140, P<0.01). Furthermore, the sensitivity, specificity, and positive and negative predictive values of type 3, 4, or 5 morphologies for predicting Helicobacter pylori positive were 94.28%, 96.66%, 98.50%, and 87.87%, respectively, correlated well with inflammation grading according to the Sydney classification (P<0.01). Conclusion. Our study suggests that gastric mucosal morphologic patterns in the Helicobacter pylori infected gastric mucosa can be reliably identified using C-NBI gastroscopy with good correlation with inflammation grading

Precancerous Gastric Lesions with Helicobacter pylori vacA+/babA2+/oipA+ Genotype Increase the Risk of Gastric Cancer

Objective. The clinical outcomes of gastric diseases such as chronic gastritis, peptic ulcer, and gastric cancer have been attributed to the interplay of virulence factors of Helicobacter pylori (H. pylori), host genetic susceptibility, and host immune responses. This study investigated the presence of cagA, vacA, iceA2, babA2, and oipA genes and their association with clinical outcomes. Methods. Chronic gastritis, atrophic gastritis, and intestinal metaplasia specimens were obtained from patients who underwent endoscopy and surgical resection between January 2017 and December 2018; specimens from gastric cancer patients treated between January 2014 and December 2018 were also added. H. pylori infection and virulence genes (cagA, vacA, iceA2, babA2, and oipA) were determined using real-time PCR. The association between H. pylori genotypes and clinical outcomes were evaluated using multivariate regression model analysis. The overall survival of gastric cancer patients was compared between genotype combinations. Results. H. pylori was positive in 166 patients with chronic gastritis, precancerous gastric lesions, and gastric cancer. The genes vacA, babA2, and oipA were most prevalent in chronic gastritis (73%), precancerous gastric lesions (62%), and gastric cancer (91%), respectively. The vacA, babA2, and oipA genes were associated with increased risk of gastric cancer (OR = 1.23; 95% CI = 1.13–3.32; P=0.033, OR = 2.64; 95% CI = 1.44–4.82, P=0.024, and OR = 2.79; 95% CI = 1.58–5.41; P=0.031, respectively). Interestingly, H. pylori vacA+/babA2+/oipA+ genotype infection was associated with increased risk of gastric cancer (OR = 3.85, 95% CI = 1.67–5.77, P=0.014). Conclusion. In this present study, we reported on the virulence genes of H. pylori infection to reveal their association with increased risk of chronic gastritis, precancerous gastric lesions, and gastric cancer. Precancerous gastric lesions with H. pylori vacA+/babA2+/oipA+ genotype increased the risk of gastric cancer

Characteristics and Risk Factors of Helicobacter pylori Associated Gastritis: A Prospective Cross-Sectional Study in Northeast Thailand

Background and Aim. Risk factors for Helicobacter pylori infection are genetic susceptibility and poor living conditions. This study aimed to investigate the Mdm2 gene, clarithromycin resistance, and possible risk factors for Helicobacter pylori infection. Methods. Risk factors and clinical characteristics were analyzed, including patient demographic data, patient income, personal history, possible source of transmission, patient symptoms, endoscopic findings, patterns of clarithromycin resistance, and patterns of Mdm2 SNIP309. Results. Ingestion of pickled fish (OR = 11.27, 95% CI = 4.31-29.45, &lt; 0.0001), salt crab (OR = 8.83, 95% CI = 1.99-39.14, &lt; 0.001), and Papaya salad (OR = 8.73, 95% CI = 4.54-16.79, &lt; 0.01). The prevalence of clarithromycin resistance was 56% (wild type, A2143/2142A, is 23.8%; mutation, A2143/2142CG, is 35.7%; wild type + mutation is 40.5%). The genetic polymorphisms of Mdm2 SNIP309 were SNIP309 T/T homozygous in 78%, SNIP309 G/T heterozygous in 19%, and SNIP309 G/G homozygous in 3%. Conclusion. Pickled fish, salt crab, and Papaya salad are positive risk factors. There was high prevalence of clarithromycin resistance. The Mdm2 SNIP309 G/G homozygous genotype might be a risk factor for gastric cancer and the fact that it is infrequent in Thailand

Improved Helicobacter pylori Eradication Rate of Tailored Triple Therapy by Adding Lactobacillus delbrueckii and Streptococcus thermophilus in Northeast Region of Thailand: A Prospective Randomized Controlled Clinical Trial

Background and Aim. To evaluate the effect of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus to Helicobacter pylori eradication in different periods of therapeutic protocol. Methods. Infected patients were randomized to one-week tailored triple therapy (esomeprazole 20 mg bid, clarithromycin 500 mg bid/metronidazole 400 mg tid if clarithromycin resistant, and amoxicillin 1000 mg bid) with placebo (group 1, n=100); one week of pretreatment with probiotics (group 2, n=100); and one week of pretreatment with probiotic followed by one week of the same probiotics after treatment (group 3, n=100). Result. PP analysis involved 292 patients, 98 in group 1, 97 in group 2, and 97 in group 3. Successful eradication was observed in 229 patients; by PP analysis, the eradication rates were significantly higher (P<0.01, 95% CI; 0.71–0.97) in group 2 and group 3 than group 1. ITT analysis eradication rates were significantly higher in group 2 and group 3 than group 1 (P<0.01 95% CI; 0.72–0.87), and there is no significant difference between the three groups (P=0.32) in terms of adverse events. Conclusion. Adding probiotics before or before and after tailored treatment can improve Helicobacter pylori eradication rates. This trial is registered with Thai Clinical Trials Registry number: TCTR20141209001