Dracunculin Inhibits Adipogenesis in Human Bone Marrow-Derived Mesenchymal Stromal Cells by Activating AMPK and Wnt/ß-Catenin Signaling.

Increased bone marrow adiposity is widely observed in patients with obesity and osteoporosis and reported to have deleterious effects on bone formation. Dracunculin (DCC) is a coumarin isolated from Artemisia spp. but, until now, has not been studied for its bioactive potential except antitrypanosomal activity. In this context, current study has reported the anti-adipogenic effect of DCC in human bone marrow-derived mesenchymal stromal cells (hBM-MSCs). DCC dose-dependently inhibited the lipid accumulation and expression of adipogenic transcription factors peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT/enhancer binding protein α (C/EBPα) in hBM-MSCs induced to undergo adipogenesis. To elucidate its action mechanism, the effect of DCC on Wnt/ß-catenin and AMPK pathways was examined. Results showed that DCC treatment activated Wnt/ß-catenin signaling pathway via AMPK evidenced by increased levels of AMPK phosphorylation and Wnt10b expression after DCC treatment. In addition, DCC treated adipo-induced hBM-MSCs exhibited significantly increased nuclear levels of ß-catenin compared with diminished nuclear PPARγ levels. In conclusion, DCC was shown to be able to hinder adipogenesis by activating the ß-catenin via AMPK, providing potential utilization of DCC as a nutraceutical against bone marrow adiposity.

Evaluation and Comparison of the Antimelanogenic Properties of Different Solvent Fractionated Cnidium japonicum Extracts in B16F10 Murine Melanoma Cells.

Cnidium japonicum is a biennial halophyte found in the salt marshes and shores of Korea and widely used in traditional Korean medicine as an ingredient. This study investigated and compared the antimelanogenic effect of solventpartitioned fractions of C. japonicum extract (CJEFs) in a B16F10 mouse melanoma cell model, focusing on tyrosinase activity and production. Melanogenesis is the process in which skin pigment melanin is produced through tyrosinase activity. Overproduction of melanin is the primary reason behind several skin disorders such as freckles, spots, and hyperpigmentation. The antimelanogenic capacity of CJEFs was initially screened by their tyrosinase inhibitory effects, prevention of dihydroxyphenylalanine (DOPA) oxidation, and suppression of melanin production. The inhibition of tyrosinase activity and DOPA oxidation by CJEFs was suggested to be related to the downregulation of microphthalmia-associated transcription factor, tyrosinase, tyrosinase-related protein-1, and tyrosinase-related protein-2, which was confirmed using mRNA and protein expression levels. Moreover, the glycogen synthase kinase 3 beta- and cyclic adenosine monophosphate response element-binding protein-related signaling pathways were inhibited by treatment with CJEFs, indicating their action mechanism. All the tested CJEFs exerted similar effects on tyrosinase activity and production. However, among those, 85% aq. MeOH was the most active fraction to suppress the signaling pathway that produces tyrosinase. These results suggest that especially the MeOH fraction of C. japonicum extract serves as a potential source of bioactive substances, with effective antimelanogenesis properties.

Myricetin 3-O-ß-D-Galactopyranoside Exhibits Potential Anti-Osteoporotic Properties in Human Bone Marrow-Derived Mesenchymal Stromal Cells via Stimulation of Osteoblastogenesis and Suppression of Adipogenesis.

Natural bioactive substances are promising lead compounds with beneficial effects on various health problems including osteoporosis. In this context, the goal of this study was to investigate the effect of myricetin 3-O-ß-D-galactopyranoside (M3G), a glycoside of a known bioactive phytochemical myricetin, on bone formation via osteogenic differentiation of human bone marrow-derived mesenchymal stromal cells (hBM-MSCs). The hBM-MSCs were induced to differentiate into osteoblasts and adipocytes in the presence or absence of M3G and the differentiation markers were analyzed. Osteoblastogenesis-induced cells treated with M3G exhibited stimulated differentiation markers: cell proliferation, alkaline phosphatase (ALP) activity, and extracellular mineralization. In terms of intracellular signaling behind the stimulatory effect of M3G, the expression of RUNX2 and osteopontin transcription factors were upregulated. It has been shown that M3G treatment increased the activation of Wnt and BMP as a suggested mechanism of action for its effect. On the other hand, M3G treatment during adipogenesis-inducement of hBM-MSCs hindered the adipogenic differentiation shown as decreased lipid accumulation and expression of PPARγ, SREBP1c, and C/EBPα, adipogenic transcription factors. In conclusion, M3G treatment stimulated osteoblast differentiation and inhibited adipocyte differentiation in induced hBM-MSCs. Osteoblast formation was stimulated via Wnt/BMP and adipogenesis was inhibited via the PPARγ pathway. This study provided necessary data for further studies to utilize the therapeutic potential of M3G against osteoporosis via regulation of bone marrow stromal cell differentiation.

Resting heart rate is associated with colorectal advanced adenoma.

BACKGROUND AND AIMS: Resting heart rate is an independent predictor of colorectal cancer (CRC) development and CRC-related mortality. However, little is known about the relationship between resting heart rate and colorectal adenoma development. We aimed to investigate this association in a population who underwent screening colonoscopy. METHODS: Among 39,021 patients who underwent both electrocardiogram and screening colonoscopy during routine health examinations at the Seoul National University Bundang Hospital, Health Promotion Center, Korea from January 2014 to July 2019, 1,344 patients had advanced adenoma. We performed 1:1 propensity score (PS) matching to establish a control group that mitigated the confounding effects of age and sex. We performed multivariate logistic regression analyses to identify the independent risk factors of advanced adenoma development. RESULTS: Resting heart rate was significantly higher in the advanced adenoma group than in the control group. The prevalence of advanced polyp increased across the quartiles of resting heart rate. Patients with higher resting heart rates were more likely to be older, smokers, and have increased blood pressure and DM and less likely to engage in active exercises than those with lower resting heart rates. Patients with higher resting heart rates had higher serum glucose, triglyceride, hemoglobin A1C, and insulin levels and lower high-density lipoprotein cholesterol levels. Patients with resting heart rate in the highest quartile (≥71 bpm) still showed significantly increased odds ratio (OR) of advanced adenoma development (OR: 1.379, 95% confidence interval: 1.099-1.731, p = 0.006). CONCLUSIONS: High resting heart rate was a meaningful independent risk factor of advanced adenoma development.

Outcomes of Minimally Invasive Oblique Lumbar Interbody Fusion in Patients with Lumbar Degenerative Disease with Rheumatoid Arthritis.

PURPOSE OF STUDY: Standard treatment protocols for lumbar degenerative lesions in the setting of rheumatoid arthritis (RA) are lacking. The purpose of this study was to evaluate the clinical and radiologic outcomes of minimally invasive oblique lumbar interbody fusion (MI-OLIF) in RA patients having degenerative lumbar spine lesions. METHODS: This was a retrospective hospital-based case series (evidence level 4). Eight patients with degenerative lumbar disease with significant back pain and neurologic claudication underwent MI-OLIFwith polyetheretherketone cage insertion and posterior pedicle screw instrumentation. The clinical outcomes were measured by the numerical rating scale (NRS) for back and leg pain and the Oswestry Disability Index (ODI), and radiologic outcomes were studied on radiographs, computed tomography, and magnetic resonance imaging. Minimum follow-up duration was 1 year. RESULTS: Mean NRS results for back and leg pain preoperatively were 6.3 and 7.1 that improved to 2.6 and 2 for back and leg pain, respectively, at last follow-up. The mean ODI scores preoperatively were 58.02 that improved to 39.06 at last follow-up. All patients had good functional outcomes, good fusion rates, and were able to continue their activities of daily living without much disability at last follow-up. CONCLUSION: MI-OLIF in patients with symptomatic lumbar spine degenerative lesions with RA seems to provide good short-term clinical and radiologic outcomes.

Association of visceral adiposity and insulin resistance with colorectal adenoma and colorectal cancer.

BACKGROUND/AIMS: To examine whether visceral adiposity serves as a risk factor for colorectal cancer (CRC) and colorectal adenomas. METHODS: Two hundred healthy subjects, 200 patients with colorectal adenoma, and 151 patients with CRC (46 with early-stage and 105 with advanced-stage cancers) were enrolled at a tertiary referral hospital. All subjects underwent colonoscopy, and had laboratory data, and computed tomography (CT) scan available for abdominal fat measurement. An abdominal CT scan taken 1 to 4 years (mean interval, 20.6 months) before the diagnosis of CRC was also available in the 42 CRC patients. RESULTS: The mean areas of visceral adipose tissue (VAT) areas in the control, adenoma, early- and advanced-stage CRC groups were 94.6, 116.8, 110.4, and 99.7 cm2 , respectively (P<0.001). The risk of adenoma positively correlated with VAT area and the visceral-to-total fat ratio (P for trend <0.01), but the risk of CRC did not (P>0.05). The risk of both adenoma and CRC positively correlated with fasting plasma glucose levels (P for trend <0.05). In patients with early-stage cancer (n=17), VAT area decreased when the CT scan at diagnosis was compared with that taken before the diagnosis of CRC, but superficial adipose tissue area did not, so visceral-to-total fat ratio significantly decreased (46.6% vs. 50.7%, respectively, P=0.018). CONCLUSIONS: VAT area is related to the risk of colorectal adenoma. However, VAT decreases from the early stages of CRC. Impaired fasting glucose has a role in colorectal carcinogenesis.

Clinical and Radiologic Outcomes of Direct Versus Indirect Decompression with Lumbar Interbody Fusion: A Matched-Pair Comparison Analysis.

OBJECTIVE: To compare the radiologic and clinical outcomes between oblique lumbar interbody fusion (OLIF) without laminectomy and minimally invasive transforaminal lumbar interbody fusion (MI-TLIF). METHODS: This was a retrospective study. Between April 2012 and January 2017, 25 patients in each of the MI-TLIF and OLIF groups were recruited as matched pairs. Clinical outcomes included visual analogue scale, Oswestry Disability Index, and satisfaction rates. Radiographic outcomes comprised disc height (DH) and fusion status. Intraoperative data and complications were collected. All patients completed the clinical and radiologic outcomes. Outcomes were compared preoperatively and postoperatively. RESULTS: Matched pairs were compared between 2 groups in terms of demographic data and preoperative measurements; less blood loss and shorter operative time were found in OLIF versus MI-TLIF (P < 0.001). The total complication rate was 36% in OLIF and 32% in MI-TLIF (P = 0.77). The outcomes of visual analogue scale and Oswestry Disability Index were significantly improved in both groups, and there was no significant difference between 2 groups. Satisfaction rates of the both groups were more than 90%. OLIF was superior to MI-TLIF with respect its capability to restore DH (P < 0.001). Earlier time of fusion was observed in OLIF (80%) compared with MI-TLIF (52%) at 6 months (P = 0.04). CONCLUSIONS: OLIF may achieve equivalent clinical and radiologic outcomes compared with MI-TLIF when the stenosis is minimal because the decompression performed is indirect. Furthermore, the OLIF shows less blood loss and shorter operative time, better restoration of DH, and earlier time to fusion than the MI-TLIF.

Omental calcification, necrotizing enterocolitis, and undescended testes after fetal abdomino-amniotic shunting performed for the management of meconium peritonitis: A case report.

Prenatal ultrasonography (US) in a 39 year-old woman revealed massive fetal ascites. A fetal abdomino-amniotic shunting procedure was performed. Subsequently, plain radiographs demonstrated diffuse gaseous distention of the bowel and multiple punctate calcifications in the left upper abdomen. Postnatal US examination showed multiple echogenic foci in the liver and the left upper abdomen, bowel wall thickening in the right-sided abdomen, and undescended testes. There was no intra-abdominal free air or loculated fluid collections. Medical management was instituted secondary to the clinical suspicion of omental calcification, necrotizing enterocolitis, and undescended testes. Follow-up US examination showed resolution of portal vein gas and bowel wall thickening. The neonate recovered fully.

Anterior transcorporeal tunnel approach for cervical myelopathy guided by CT-based intraoperative spinal navigation: Technical note.

Cervical spondylotic myelopathy (CSM) is the most common degenerative disease of the spine in elderly patients. Although there are several surgical options for the treatment of this disease, minimally invasive techniques for localized decompression of the true clinical generator of the patient's disease has revolutionized spinal surgery recently. This article presents an alternate application of the anterior transcorporeal tunnel approach in combination with computed tomography (CT)-based intraoperative navigation in the treatment of patients with cervical myelopathy secondary to spondylosis. Three clinical cases are used as examples to demonstrate how this procedure can be used to decompress a localized target in cervical spondylotic myelopathy. Clinical images of each case are shown. Cervical decompression was successfully achieved in all three patients, with the improvement of preoperative symptoms. The anterior transcorporeal tunnel approach combined with the use of CT-based intraoperative spinal navigation may be a feasible minimally invasive procedure for the treatment of cervical spondylotic myelopathy in selected cases.

Comparison of Changes in the Interstitial Cells of Cajal and Neuronal Nitric Oxide Synthase-positive Neuronal Cells With Aging Between the Ascending and Descending Colon of F344 Rats.

BACKGROUND/AIMS: Neuronal degeneration and changes in interstitial cells of Cajal (ICCs) are important mechanisms of age-related constipation. This study aims to compare the distribution of ICCs and neuronal nitric oxide synthase (nNOS) with regard to age-related changes between the ascending colon (AC) and descending colon (DC) in 6-, 31-, and 74-week old and 2-year old male Fischer-344 rats. METHODS: The amount of fecal pellet and the bead expulsion times were measured. Fat proportion in the muscle layer of the colon was analyzed by hematoxylin and eosin staining. Proto-oncogene receptor tyrosine kinase (KIT) and neuronal nitric oxide synthase (nNOS) expression were analyzed with Western blotting and immunohistochemistry. Isovolumetric contractile measurements and electrical field stimulation were used to assess smooth muscle contractility. RESULTS: Colon transit and bead expulsion slowed with senescence. Fat in the muscle layer accumulated with age in the AC, but not in the DC. The proportion of KIT-immunoreactive ICCs in the submucosal and myenteric plexus was higher in the DC than in the AC, and it declined with age, especially in the AC. In contrast, the proportion of NOS-immunoreactive neurons in the myenteric plexus was higher in the AC than in the DC, and both decreased in older rats. Nitric oxide levels declined with age in the DC. Muscle strip experiments showed that the inhibitory response mediated by nitric oxide in the circular direction of the DC was reduced in 2-year old rats. CONCLUSION: The AC and DC differ in their distribution of ICCs and nNOS, and age-related loss of nitrergic neurons more severely affects the DC than the AC.

Analysis of Gastric Body Microbiota by Pyrosequencing: Possible Role of Bacteria Other Than Helicobacter pylori in the Gastric Carcinogenesis.

BACKGROUND: Gastric microbiota along with Helicobacter pylori (HP) plays a key role in gastric disease. The aim of our study is to investigate the difference of human gastric microbiota between antrum and body according to disease (control vs. gastric cancer) and HP status. METHODS: Each antrum and body biopsy was collected from 12 subjects at Seoul National University Bundang Hospital. Gastric microbiota was analyzed by bar-coded 454 pyrosequencing of the 16S rRNA gene. Twelve subjects consisted of HP-negative control (n = 2), HP-negative cancer (n = 2), HP-positive control (n = 3), and HP-positive cancer (n = 5). The analysis was focused on non-HP urease-producing bacteria (UB) and non-HP nitrosating or nitroreducing bacteria (NB) between antrum and body. RESULTS: Gastric body samples showed higher diversity compared to gastric antrum mucosa samples but there was no significant difference. The mean of operational taxonomic units was higher in HP(-) cancer than HP(+) cancer (antrum, 273.5 vs. 228.2, P = 0.439; body, 585.5 vs. 183.2, P = 0.053). The number of non-HP UB and non-HP NB was higher in HP(-) cancer groups than the others. These differences were more pronounced in the body (P = 0.051 and P = 0.081, respectively). Analysis of overlap of non-HP UB and non-HP NB revealed the higher composition of Streptococcus pseudopneumoniae, S. parasanguinis, and S. oralis in HP(-) cancer groups than the others, only in the body (P = 0.030) but not in the antrum (P = 0.123). CONCLUSIONS: Higher diversity and higher composition of S. pseudopneumoniae, S. parasanguinis, and S. oralis in HP(-) cancer group than the other groups in the body suggest that analysis of microbiota from body mucosa could be beneficial to identify a role of non-HP bacteria in the gastric carcinogenesis.

Minimally Invasive Transforaminal Lumbar Interbody Fusion: A Comparison Study Based on End Plate Subsidence and Cystic Change in Individuals Older and Younger than 65 Years.

OBJECTIVE: To compare the outcomes between patients older and younger than 65 years who underwent single-level minimally invasive transforaminal interbody fusion (MI-TLIF) surgery. METHODS: This study is a retrospective analysis of 76 patients who underwent MI-TLIF between April 2012 and June 2016. Group A consisted of 35 patients (<65 years) and group B consisted of 41 patients (≥65 years). Intraoperative data were recorded. The evaluation of clinical outcomes was based on the visual analog scale for back and leg pain and the Oswestry Disability Index. Radiologic outcomes including cage subsidence, end plate cyst formation, and fusion rate were assessed. RESULTS: The mean age of the study subjects was 65.3 years, and the mean duration of follow-up was 18.98 months. Group B had a higher rate of comorbidities compared with group A (90.24% vs. 57.14%, respectively; P < 0.05). There was no statistically significant difference in the rate of complications between the groups (group A, 14.29%; group B, 17.07%). Clinical outcomes significantly improved in both groups postoperatively (P < 0.05). Although bony fusion in group A was slightly higher than that in group B, the fusion rate was not statistically different according to age. There were no statistically significant differences in the rates of cage subsidence or positive cyst sign between the groups. CONCLUSIONS: MI-TLIF presented similar safeness and acceptable outcomes and complication rate in both groups. Cyst formation may be aggravated by cage subsidence, because cage subsidence was a useful potential predictor of cyst formation.

Pantoprazole Does Not Reduce the Antiplatelet Effect of Clopidogrel: A Randomized Controlled Trial in Korea.

BACKGROUND/AIMS: Concerns that proton pump inhibitors (PPIs) diminish the efficacy of clopidogrel could hamper the appropriate prescription of PPIs. We evaluated the influence of pantoprazole on the antiplatelet effect of clopidogrel compared with ranitidine, which is regarded as safe, after stratification of the population according to the presence of a cytochrome (CYP) 2C19 polymorphism in Korea. METHODS: Forty patients who underwent dual antiplatelet therapy were randomized to receive pantoprazole (n=20) or ranitidine (n=20). Platelet aggregation was evaluated by impedance aggregometry at baseline (D0) and 8 days after acid-lowering treatments (D9). CYP2C19 was genotyped by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: After co-treatment, the percentage of clopidogrel low-response was 11.1% (2/18) in the pantoprazole group and 10.5% (2/19) in the ranitidine group (p=0.954). The impedance values with adenosine diphosphate stimulus after acid-lowering treatments did not significantly differ between the two groups. In a multiple regression analysis, only ST-elevation myocardial infarction was marginally associated with a reduced antiplatelet effect (odds ratio, 12.07; 95% confidence interval, 0.84 to 173.78). However, pantoprazole use did not affect the antiplatelet effect after correction for the CYP2C19 polymorphism. CONCLUSIONS: This study showed that pantoprazole does not increase platelet aggregation in patients receiving dual antiplatelet therapy (ClinicalTrials.gov number: NCT02733640).

Erratum: Analysis of Gastric Body Microbiota by Pyrosequencing: Possible Role of Bacteria Other Than Helicobacter pylori in the Gastric Carcinogenesis.

[This corrects the article on p. 115 in vol. 22, PMID: 28698866.].

PMK-S005 Alleviates Age-Related Gastric Acid Secretion, Inflammation, and Oxidative Status in the Rat Stomach.

BACKGROUND/AIMS: The aim of this study was to evaluate the effect of the synthetic S-allyl-L-cysteine (SAC) PMK-S005 on gastric acid secretion, inflammation, and antioxidant enzymes in aging rats. METHODS: The rats were divided into four groups at 31 weeks of age and were continuously fed a diet containing a vehicle control, PMK-S005 (5 or 10 mg/kg), or lansoprazole (5 mg/kg). Gastric acid secretion and connective tissue thickness of the lamina propria were evaluated at 74 weeks and 2 years of age. Tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, and COX-2 levels were measured by using enzyme-linked immunosorbent assays (ELISAs) or Western blot assays. Levels of antioxidant enzymes, including heme oxyganase 1 (HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO-1), were also measured. RESULTS: As the rats aged, gastric acid secretion significantly decreased, and the connective tissue of the lamina propria increased. However, 74-week-old rats in the PMK-S005 group exhibited greater levels of gastric acid secretion than those of the control and lansoprazole groups. The increase of TNF-α, IL-1ß, and COX- 2 expression in 74-week and 2-year-old control rats were inhibited by PMK-S005. In addition, the decrease in HO-1 and NQO-1 protein expression that occurred with aging was inhibited by PMK-S005 in the 74-week-old rats. CONCLUSIONS: These results suggest that PMK-S005 has therapeutic potential as an antiaging agent to ameliorate age-related gastric acid secretion, inflammation, and oxidative stress in the stomach.

Comparison of Gastric Microbiota Between Gastric Juice and Mucosa by Next Generation Sequencing Method.

BACKGROUND: Not much is known about the role of gastric microbiota except for Helicobacter pylori in human health and disease. In this study, we aimed to detect human gastric microbiota in both gastric mucosa and gastric juice by barcoded 454-pyrosequencing of the 16S rRNA gene and to compare the results from mucosa and juice. METHODS: Gastric biopsies and stomach juices were collected from 4 subjects who underwent standard endoscopy at Seoul National University Bundang Hospital. Gastric microbiota of antral mucosa, corpus mucosa samples, and gastric fluids were analyzed by barcoded 454-pyrosequencing of the 16S rRNA gene. The analysis focused on bacteria, such as H. pylori and nitrosating or nitrate-reducing bacteria. RESULTS: Gastric fluid samples showed higher diversity compared to that of gastric mucosa samples. The mean of operational taxonomic units was higher in gastric fluid than in gastric mucosa. The samples of gastric fluid and gastric mucosa showed different composition of phyla. The composition of H. pylori and Proteobacteria was higher in mucosa samples compared to gastric fluid samples (H. pylori, 66.5% vs. 3.3%, P = 0.033; Proteobacteria, 75.4% vs. 26.3%, P = 0.041), while Actinobacteria, Bacteroidetes, and Firmicutes were proportioned relatively less in mucosa samples than gastric fluid. However there was no significant difference. (Actinobacteria, 3.5% vs. 20.2%, P = 0.312; Bacteroidetes, 6.0% vs. 14.8%, P = 0.329; Firmicutes, 12.8% vs. 33.4%, P = 0.246). CONCLUSIONS: Even though these samples were small, gastric mucosa could be more effective than gastric fluid in the detection of meaningful gastric microbiota by pyrosequencing.

Analysis of Gastric Microbiota by Pyrosequencing: Minor Role of Bacteria Other Than Helicobacter pylori in the Gastric Carcinogenesis.

BACKGROUND: Little is known about the role of gastric microbiota except for Helicobacter pylori (HP) in human health and disease. We compared the differences of human gastric microbiota according to gastric cancer or control and HP infection status and assessed the role of bacteria other than HP. METHODS: Gastric microbiota of 63 antral mucosal and 18 corpus mucosal samples were analyzed by bar-coded 454 pyrosequencing of the 16S rRNA gene. Antral samples were divided into four subgroups based on HP positivity in pyrosequencing and the presence of cancer. The analysis was focused on bacteria other than HP, especially nitrosating or nitrate-reducing bacteria (NB). The changes of NB in antral mucosa of 16 subjects were followed up. RESULTS: The number of NB other than HP (non-HP-NB) was two times higher in the cancer groups than in the control groups, but it did not reach statistical significance. The number of non-HP-NB tends to increase over time, but this phenomenon was prevented by HP eradication in the HP-positive control group, but not in the HP-positive cancer group. CONCLUSION: We could not find the significant role of bacteria other than HP in the gastric carcinogenesis.

Comparison between Resectable Helicobacter pylori-Negative and -Positive Gastric Cancers.

BACKGROUND/AIMS: Controversy exists regarding the characteristics of Helicobacter pylori infection-negative gastric cancer (HPIN-GC). The aim of this study was to evaluate clinicopathologic features of HPIN-GC compared to H. pylori infection-positive gastric cancer (HPIP-GC) using a comprehensive analysis that included genetic and environmental factors. METHODS: H. pylori infection status of 705 resectable gastric cancer patients was determined by the rapid urease test, testing for anti-H. pylori antibodies, histologic analysis and culture of gastric cancer tissue samples, and history of H. pylori eradication. HPIN-GC was defined as gastric cancer that was negative for H. pylori infection based on all five methods and that had no evidence of atrophy in histology or serology. RESULTS: The prevalence of HPIN-GC was 4% (28/705). No significant differences with respect to age, sex, smoking, drinking, family history of gastric cancer or obesity were observed between the two groups. HPIN-GC tumors were marginally more likely to involve the cardia (14.3% for HPIN-GC vs 5.3% for HPIP-GC, p=0.068). The Lauren classification, histology, and TNM stage did not differ according to H. pylori infection status. Microsatellite instability was not different between the two groups, but p53 overexpression in HPIN-GC was marginally higher than in HPIP-GC (56.0% for HPIN-GC vs 37.0% for HPIP-GC, p=0.055). CONCLUSIONS: The prevalence of HPIN-GC was extremely low, and its clinicopathologic characteristics were similar to HPIP-GC.

Clinicopathologic and molecular features associated with patient age in gastric cancer.

AIM: To compare characteristics and prognosis of gastric cancer based on age. METHODS: A retrospective study was conducted on clinical and molecular data from patients (n = 1658) with confirmed cases of gastric cancer in Seoul National University Bundang Hospital (Seoul, South Korea) from 2003 to 2010 after exclusion of patients diagnosed with lymphoma, gastrointestinal stromal tumor, and metastatic cancer in the stomach. DNA was isolated from tumor and adjacent normal tissue, and a set of five markers was amplified by polymerase chain reaction to assess microsatellite instability (MSI). MSI was categorized as high, low, or stable if ≥ 2, 1, or 0 markers, respectively, had changed. Immunohistochemistry was performed on tissue sections to detect levels of expression of p53, human epidermal growth factor receptor (HER)-2, and epidermal growth factor receptor. Statistical analysis of clinical and molecular data was performed to assess prognosis based on the stratification of patients by age (≤ 45 and > 45 years). RESULTS: Among the 1658 gastric cancer patients, the number of patients with an age ≤ 45 years was 202 (12.2%; 38.9 ± 0.4 years) and the number of patients > 45 years was 1456 (87.8%; 64.1 ± 0.3 years). Analyses revealed that females were predominant in the younger group (P < 0.001). Gastric cancers in the younger patients exhibited more aggressive features and were at a more advanced stage than those in older patients. Precancerous lesions, such as atrophic gastritis and intestinal metaplasia, were observed less frequently in the older than in the younger group (P < 0.001). Molecular characteristics, including overexpression of p53 (P < 0.001), overexpression of HER-2 (P = 0.006), and MSI (P = 0.006), were less frequent in gastric cancer of younger patients. Cancer related mortality was higher in younger patients (P = 0.048), but this difference was not significant after adjusting for the stage of cancer. CONCLUSION: Gastric cancer is distinguishable between younger and older patients based on both clinicopathologic and molecular features, but stage is the most important predictor of prognosis.

An Appropriate Cutoff Value for Determining the Colonization of Helicobacter pylori by the Pyrosequencing Method: Comparison with Conventional Methods.

BACKGROUND: Sequencing of 16S ribosomal RNA (rRNA) gene has improved the characterization of microbial communities. It enabled the detection of low abundance gastric Helicobacter pylori sequences even in subjects that were found to be H. pylori negative with conventional methods. The objective of this study was to obtain a cutoff value for H. pylori colonization in gastric mucosa samples by pyrosequencing method. MATERIALS AND METHODS: Gastric mucosal biopsies were taken from 63 subjects whose H. pylori status was determined by a combination of serology, rapid urease test, culture, and histology. Microbial DNA from mucosal samples was amplified by PCR using universal bacterial primers. 16S rDNA amplicons were pyrosequenced. ROC curve analysis was performed to determine the cutoff value for H. pylori colonization by pyrosequencing. In addition, temporal changes in the stomach microbiota were observed in eight initially H. pylori-positive and eight H. pylori-negative subjects at a single time point 1-8 years later. RESULTS: Of the 63 subjects, the presence of H. pylori sequences was detected in all (28/28) conventionally H. pylori-positive samples and in 60% (21/35) of H. pylori-negative samples. The average percent of H. pylori reads in each sample was 0.67 ± 1.09% in the H. pylori-negative group. Cutoff value for clinically positive H. pylori status was approximately 1.22% based on ROC curve analysis (AUC = 0.957; p < .001). Helicobacter pylori was successfully eradicated in five of seven treated H. pylori-positive subjects (71.4%), and the percentage of H. pylori reads in these five subjects dropped from 1.3-95.18% to 0-0.16% after eradication. CONCLUSION: These results suggest that the cutoff value of H. pylori sequence percentage for H. pylori colonization by pyrosequencing could be set at approximately 1%. It might be helpful to analyze gastric microbiota related to H. pylori sequence status.