Overview of Citrin Deficiency：SLC25A13 Mutations and the Frequency

Citrin deficiency,autosomal recessive disorder,caused by mutation of SLC25A13 gene on chromosome 7q21.3 has two major phenotypes:neonatal intrahepatic cholestatic hepatitis(NICCD)and adult-onset type Ⅱ citrullinemia(CTLN2).So far,we have identified 52 SLC25A13 mutations and diagnosed the patients not only in Japan(166 CTLN2 and 238 NICCD) but also in other countries.We have detected 76 Chinese,13 Korean and 15 Vietnamese patients with the same mutations as Japanese,and 13 patients(from Israel,UK,USA or Czech)with mutations different from those found in Japanese,indicating a wide distribution of citrin deficiency.DNA diagnoses of 13 known SLC25A13 mutations revealed that the carrier frequency was high in East Asian populations:Chinese(73/4 600=1/63),Japanese(21/1 372=1/65) and Korean(25/2 690=1/108),suggesting that near by 100 000 East Asians are homozygotes.It is important to find out patients with citrin deficiency,to treat them,and to prevent onset of severe CTLN2.

Different effects of ghrelin, des-acyl ghrelin and obestatin on gastroduodenal motility in conscious rats

Three peptides, ghrelin, des-acyl ghrelin and obestatin are derived from a common prohormone, preproghrelin by posttranslational processing, originating from endocrine cells in the stomach. To examine the effects of these peptides, we applied the manometric mea- surement of gastrointestinal motility in freely moving conscious rat models. Ghrelin exerts stimulatory ef- fects on the motility of antrum and duodenum in both fed and fasted state of animals. Des-acyl ghrelin exerts inhibitory effects on the motility of antrum, but not on the motility of duodenum in the fasted state of ani- mals. Obestatin exerts inhibitory effects on the motility of antrum and duodenum in the fed state, but not in the fasted state of animals. NPY Y2 or Y4 receptors in the brain may mediate the action of ghrelin, CRF type 2 receptors in the brain mediate the action of des-acyl ghrelin, whereas CRF type 1 and type 2 receptors in the brain mediate the action of obestatin. Vagal afferent pathways might be involved in the action of ghre- lin, but not involved in the action of des-acyl ghrelin, whereas vagal afferent pathways might be partially involved in the action of obestatin.

Polymorphisms of alcohol dehydrogenase 2 and aldehyde dehydrogenase 2 and colorectal cancer risk in Chinese males

AIM: To evaluate the relationship between drinking and polymorphisms of alcohol dehydrogenase 2 (ADH2) and/or aldehyde dehydrogenase 2 (ALDH2) for risk of colorectal cancer (CRC) in Chinese males. METHODS: A case-control study was conducted in 190 cases and 223 population-based controls. ADH2 Arg47His (G-A) and ALDH2 Glu487Lys (G-A)genotypes were identified by PCR and denaturing high-performance liquid chromatography (DHPLC). Information on smoking and drinking was collected and odds ratio (OR) was estimated. RESULTS: The ADH2 A/A and ALDH2 G/G genotypes showed moderately increased CRC risk. The age- and smoking-adjusted OR for ADH2 A/A relative to G/A and G/G was 1.60 (95% CI=1.08-2.36), and the adjusted OR for ALDH2 G/G relative to G/A and A/A was 1.79 (95% CI=1.19-2.69). Signif icant interactions between ADH2, ALDH2 and drinking were observed. As compared to the subjects with ADH2 G and ALDH2 A alleles, those with ADH2 A/A and ALDH2 G/G genotypes had a signif icantly increased OR (3.05, 95% CI= 1.67-5.57). The OR for CRC among drinkers with the ADH2 A/A genotype was increased to 3.44 (95% CI= 1.84-6.42) compared with non-drinkers with the ADH2 G allele. The OR for CRC among drinkers with the ALDH2 G/G genotype was also increased to 2.70 (95% CI= 1.57-4.66) compared with non-drinkers with the ALDH2 A allele. CONCLUSION: Polymorphisms of the ADH2 and ALDH2 genes are significantly associated with CRC risk. There are also signifi cant gene-gene and gene- environment interactions between drinking and ADH2 and ALDH2 polymorphisms regarding CRC risk in Chinese males.

PiggyBac transposon-mediated gene delivery efficiently generates stable transfectants derived from cultured primary human deciduous tooth dental pulp cells(HDDPCs) and HDDPC-derived iPS cells

The ability of human deciduous tooth dental pulp cells（HDDPCs） to differentiate into odontoblasts that generate mineralized tissue holds immense potential for therapeutic use in the field of tooth regenerative medicine. Realization of this potential depends on efficient and optimized protocols for the genetic manipulation of HDDPCs. In this study, we demonstrate the use of a Piggy Bac（PB）-based gene transfer system as a method for introducing nonviral transposon DNA into HDDPCs and HDDPC-derived inducible pluripotent stem cells. The transfection efficiency of the PB-based system was significantly greater than previously reported for electroporation-based transfection of plasmid DNA. Using the neomycin resistance gene as a selection marker, HDDPCs were stably transfected at a rate nearly 40-fold higher than that achieved using conventional methods. Using this system, it was also possible to introduce two constructs simultaneously into a single cell. The resulting stable transfectants, expressing td Tomato and enhanced green fluorescent protein, exhibited both red and green fluorescence. The established cell line did not lose the acquired phenotype over three months of culture. Based on our results, we concluded that PB is superior to currently available methods for introducing plasmid DNA into HDDPCs. There may be significant challenges in the direct clinical application of this method for human dental tissue engineering due to safety risks and ethical concerns. However, the high level of transfection achieved with PB may have significant advantages in basic scientific research for dental tissue engineering applications, such as functional studies of genes and proteins. Furthermore, it is a useful tool for the isolation of genetically engineered HDDPC-derived stem cells for studies in tooth regenerative medicine.

Computed tomography and magnetic resonance imaging features of lipid-rich neuroendocrine tumors of the pancreas

AIM:To clarify the computed tomography(CT) and magnetic resonance imaging(MRI) characteristics of lipid-rich pancreatic neuroendocrine tumors(Pan NETs).METHODS:Enhanced CT and MRI performed before pancreatectomy in 29 patients with 34 histologicallyconfirmed Pan NETs was retrospectively reviewed. Tumor attenuation on CT and signal intensities on conventional(T1- and T2-weighted) and chemical shift MRI were qualitatively analyzed and compared alongside adipose differentiation-related protein(ADRP) immunostaining(ADRP-positive:lipid-rich; ADRP-negative:non-lipid-rich) results using Fisher's exact test or the Mann-Whitney U test. Signal intensity index on chemical shift MRI was quantitatively assessed.RESULTS:There were 15 lipid-rich Pan NETs(44.1%) in 12 patients(41.4%). Tumor attenuation during the early,portal venous,and delayed phases of enhanced CT(P = 0.888,0.443,and 0.359,respectively) and signal intensities on conventional MRI(P = 0.698 and 0.798,respectively) were not significantly differentbetween lipid-rich and non-lipid-rich Pan NETs. Four of the 15 lipid-rich Pan NETs exhibited high signal intensity on subtraction chemical shift MRI,and the association of high signal intensity on subtraction imaging with lipid-rich Pan NETs was significant(4 of 15 lipid-rich Pan NETs,26.73%,vs 0 of 19 non-lipid-rich Pan NETs,0%,P = 0.029). Lipid-rich Pan NETs showed a significantly higher signal intensity index than non-lipidrich Pan NETs(0.6% ± 14.1% vs-10.4% ± 14.4%,P = 0.004). Eight of 15 lipid-rich Pan NETs,vs 0 of 19 nonlipid-rich Pan NETs,had positive signal intensity index values in concordance with lipid contents.C O N C L U S I O N :C T c o n t ra s t e n h a n c e m e n t a n d conventional MR signal intensities are similar in lipidrich and non-lipid-rich Pan NETs. Chemical shift MRI can demonstrate cytoplasmic lipids in Pan NETs.

Efficacy of various endoscopic modalities in detecting dysplasia in ulcerative colitis:A systematic review and network meta-analysis

BACKGROUND Longstanding ulcerative colitis(UC)is associated with an increased risk of colonic neoplasia.Various endoscopic modalities,such as chromoendoscopy(CE),narrow band imaging(NBI)and random biopsy have been introduced for surveillance,however,there exists a paucity of direct comparisons between them.We aimed to conduct a network meta-analysis of randomized controlled trials(RCTs)performed for surveillance of neoplasia in UC.AIM To provide a comparative evaluation of the efficacy of the above-mentioned various modalities.METHODS We searched MEDLINE/PubMed,Web of Science,Embase,Google Scholar and Cochrane Central Registry through May 2016 for RCTs evaluating the efficacy of endoscopic modalities for surveillance of neoplasia in UC.The primary outcomes of interest were dysplasia(low-or high-grade)detection rates per biopsy and per patient,and dysplasia numbers per patient.Studies were simultaneously analyzed using a random-effects network meta-analysis under the Bayesian framework to identify the modality with the highest dysplasia detection rate.The best ranking probability for the dysplasia detection rate was analyzed by surface under the cumulative ranking(SUCRA)technique.RESULTS Six prospective RCTs of a total 1038 patients were identified.We identified 4 different modalities;white light(WL)high definition(HD)or standard definition(SD),CE HD,and NBI HD.For dysplasia per biopsy,direct meta-analysis showed superiority of NBI HD over WL HD and CE HD over WL SD.Network meta-analysis demonstrated the rank order of best modality as NBI HD,CE HD,WL HD and WL SD with close SUCRA scores of the first two.For dysplasia per patient,direct meta-analyses showed equivocal results between each modality.Network meta-analysis demonstrated the rank order of best modality as WL HD,NBI HD,CE HD and WL SD with small differences of the SUCRA score among the first two.For dysplasia numbers per patient,direct meta-analysis showed superiority of CE HD over WL SD.Network meta-analysis demonstrated the rank order of best modality as WL HD,NBI HD,CE HD,and WL SD with small differences of the SUCRA score among the first three.CONCLUSION We demonstrated that there were small differences among WL HD,NBI HD,and CE HD,while WL SD was inferior,in detecting dysplasia in UC.

Involvement of A5/A7 noradrenergic neurons and B2 serotonergic neurons in nociceptive processing:a fiber photometry study

In the central nervous system,the A6 noradrenaline(NA)and the B3 serotonin(5-HT)cell groups are well-recognized players in the descending antinociceptive system,while other NA/5-HT cell groups are not well characterized.A5/A7 NA and B25-HT cells project to the spinal horn and form descending pathways.We recorded G-Ca MP6 green fluorescence signal intensities in the A5/A7 NA and the B25-HT cell groups of awake mice in response to acute tail pinch stimuli,acute heat stimuli,and in the context of a non-noxious control test,using fiber photometry with a calcium imaging system.We first introduced G-Ca MP6 in the A5/A7 NA or B25-HT neuronal soma,using transgenic mice carrying the tetracycline-controlled transactivator transgene under the control of either a dopamineβ-hydroxylase or a tryptophan hydroxylase-2 promoters and by the site-specific injection of adeno-associated virus(AAV-Tet O(3 G)-G-Ca MP6).After confirming the specific expression patterns of G-Ca MP6,we recorded G-Ca MP6 green fluorescence signals in these sites in awake mice in response to acute nociceptive stimuli.G-Ca MP6 fluorescence intensity in the A5,A7,and B2 cell groups was rapidly increased in response to acute nociceptive stimuli and soon after,it returned to baseline fluorescence intensity.This was not observed in the non-noxious control test.The results indicate that acute nociceptive stimuli rapidly increase the activities of A5/A7 NA or B25-HT neurons but the non-noxious stimuli do not.The present study suggests that A5/A7 NA or B25-HT neurons play important roles in nociceptive processing in the central nervous system.We suggest that A5/A7/B2 neurons may be new therapeutic targets.All performed procedures were approved by the Institutional Animal Use Committee of Kagoshima University(MD17105)on February 22,2018.

Microbial and enzyme technology: An efficient and convenient method for MiniPrep analysis of recombinant plasmids

Minipreparation (MiniPrep) analysis is an essential step for obtaining a recombinant plasmid that carries a DNA insert containing a gene of interest. The most commonly used method for this involves cultivation of transformed Escherichia coli (E. coli) in liquid medium, brief centrifugation for precipitation of bacterial pellets, and subsequent lysis of the pellets. This process is time-consuming and laborious, especially when the sample number is high. Here, we describe a more convenient method for MiniPrep analysis that utilizes solid medium-based cultivation of bacteria.

Clear cell carcinoma of salivary gland type presenting as an endotracheal polypoid tumor

A 66-year-old woman presented with a 2-year history of asthma has progression of dyspnea within a week, and a computed tomography (CT) revealed an endotracheal tumor at level of aortic arch. The resected tumor, measured by 18 × 12 × 10 mm, has histologically malignant features including frank invasion with desmoplastic stroma and foci of necrosis. Presence of glycogen and mucin was confirmed by periodic acid-Schiff (PAS) and diastase PAS staining. Immunohistochemically, the tumor cells were diffusely positive for cytokeratin (CK) 7 and AE1/3, focally positive for CK5/6 and CK14, but negative for CK20. Thyroid transcription factor-1 (TTF-1), Napsin A, and SP-A, indicating lung parenchymal origin were all negative. Other muscular markers, neuroendocrine markers, renal cell carcinoma markers, and markers related to perivascular epithelioid cell tumors were all negative. Postoperative systemic examination suggests no primary tumor outside the lung. The follow up was excellent being recurrence free for 14 months in spite of the positive surgical margin by the tumor cells. This is a first report ?of rare neoplasm, clear cell carcinoma of salivary gland type, presenting as an endotracheal mass.

Gastric cancer risk in relation to tobacco use and alcohol drinking in Kerala, India-Karunagappally cohort study

AIM: To assess the risk of gastric cancer(GC) in relation to tobacco use and alcohol drinking in the Karunagappally cohort in Kerala, South India.METHODS: This study examined the association of tobacco use and alcohol drinking with GC incidence among 65553 men aged 30-84 in the Karunagappally cohort. During the period from 1990-2009, 116 GC cases in the cohort were identified as incident cancers. These cases were identified from the populationbased cancer registry. Information regarding risk factors such as socioeconomic factors and tobacco and alcohol habits of cohort members were collected from the database of the baseline survey conducted during 1990-1997. The relative risks(RRs) and the corresponding 95% confidence intervals(95%CIs) fortobacco use were obtained from Poisson regression analysis of grouped survival data, considering age, follow-up period, occupation and education.RESULTS: Bidi smoking was associated with GC risk(P = 0.042). The RR comparing current versus never smokers was 1.6(95%CI: 1.0-2.5). GC risk was associated with the number of bidis smoked daily(P = 0.012) and with the duration of bidi smoking(P = 0.036). Those who started bidi smoking at younger ages were at an elevated GC risk; the RRs for those starting bidi smoking under the age of 18 and ages 18-22 were 2.0(95%CI: 1.0-3.9) and 1.8(95%CI: 1.1-2.9), respectively, when their risks were compared with lifetime non-smokers of bidis. Bidi smoking increased the risk of GC among never cigarette smokers more evidently(RR = 2.2; 95%CI: 1.3-4.0). GC risk increased with the cumulative amount of bidi smoking, which was calculated as the number of bidis smoked per day x years of smoking(bidi-year; P = 0.017). Cigarette smoking, tobacco chewing or alcohol drinking was not significantly associated with GC risk. CONCLUSION: Among a male cohort in South India, gastric cancer risk increased with the number and duration of bidi smoking.

Biochemical characteristics of neonatal cholestasis induced by citrin deficiency

AIM:To explore differences in biochemical indices between neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) and that with other etiologies. METHODS:Patients under 6 mo of age who were referred for investigation of conjugated hyperbiliru-binaemia from June 2003 to December 2010 were eligible for this study. After excluding diseases affecting the extrahepatic biliary system, all patients were screened for the two most common SLC25A13 mutations; the coding exons of the entire SLC25A13 gene was sequenced and Western blotting of citrin protein performed in selected cases. Patients in whom homo-zygous or compound heterozygous SLC25A13 mutation and/or absence of normal citrin protein was detected were defined as having NICCD. Cases in which no specific etiological factor could be ascertained after a com-prehensive conjugated hyperbilirubinaemia work-up were defined as idiopathic neonatal cholestasis (INC). Thirty-two NICCD patients, 250 INC patients, and 39 infants with cholangiography-confirmed biliary atresia (BA) were enrolled. Laboratory values at their first visit were abstracted from medical files and compared. RESULTS:Compared with BA and INC patients, the NICCD patients had significantly higher levels of total bile acid (TBA) [all measures are expressed as median (inter-quartile range):178.0 (111.2-236.4) μmol/L in NICCD vs 112.0 (84.9-153.9) μmol/L in BA and 103.0 (70.9-135.3) μmol/L in INC, P = 0.0001]. The NICCD patients had significantly lower direct bilirubin [D-Bil 59.6 (43.1-90.9) μmol/L in NICCD vs 134.0 (115.9-151.2) μmol/L in BA and 87.3 (63.0-123.6) μmol/L in INC, P = 0.0001]; alanine aminotransferase [ALT 34.0 (23.0-55.0) U/L in NICCD vs 108.0 (62.0-199.0) U/L in BA and 84.5 (46.0-166.0) U/L in INC, P = 0.0001]; aspartate aminotransferase [AST 74.0 (53.5-150.0) U/L in NICCD vs 153.0 (115.0-239.0) U/L in BA and 130.5 (81.0-223.0) U/L in INC, P = 0.0006]; albumin [34.9 (30.7-38.2) g/L in NICCD vs 38.4 (36.3-42.2) g/L in BA and 39.9 (37.0-42.3) g/L in INC, P = 0.0001]; glucose [3.2 (2.0-4.4) mmol/L in NICCD vs 4.1 (3.4-5.1) mmol/L in BA and 4.0 (3.4-4.6) mmol/L in INC, P = 0.0014] and total cholesterol [TCH 3.33 (2.97-4.00) mmol/L in N ICCD vs 4.57 (3.81-5.26) mmol/L in BA and 4.00 (3.24-4.74) mmol/L in INC, P = 0.0155] levels. The D-Bil to total bilirubin (T-Bil) ratio was significantly lower in NICCD patients [all measures are expressed as median (inter-quartile range):0.54 (0.40-0.74)] than that in BA patients [0.77 (0.72-0.81), P = 0.001] and that in INC patients [0.74 (0.59-0.80), P = 0.0045]. A much higher AST/ALT ratio was found in NICCD patients [2.46 (1.95-3.63)] compared to BA patients [1.38 (0.94-1.97), P = 0.0001] and INC patients [1.48 (1.10-2.26), P = 0.0001]. NICCD patients had significantly higher TBA/D-Bil ratio [3.36 (1.98-4.43) vs 0.85 (0.72-1.09) in BA patients and 1.04 (0.92-1.14) in INC patients, P = 0.0001], and TBA/TCH ratio [60.7 (32.4-70.9) vs 24.7 (19.8-30.2) in BA patients and 24.2 (21.4-26.9) in INC patients, P = 0.0001] compared to the BA and INC groups. CONCLUSION:NICCD has significantly different bio- chemical indices from BA or INC. TBA excretion in NICCD appeared to be more severely disturbed than that of bilirubin and cholesterol.

Gd-EOB-DTPA-enhanced magnetic resonance imaging features of hepatic hemangioma compared with enhanced computed tomography

AIM:To clarify features of hepatic hemangiomas on gadolinium-ethoxybenzyl-diethylenetriaminpentaacetic acid (Gd-EOB-DTPA)-enhanced magnetic resonance imaging (MRI) compared with enhanced computed tomography (CT). METHODS:Twenty-six patients with 61 hepatic hem- angiomas who underwent both Gd-EOB-DTPA-enhanced MRI and enhanced CT were retrospectively reviewed. Hemangioma appearances (presence of peripheral nodular enhancement, central nodular enhancement, diffuse homogenous enhancement, and arterioportal shunt during the arterial phase, fill-in enhancement during the portal venous phase, and prolonged enhancement during the equilibrium phase) on Gd-EOB-DTPA-enhanced MRI and enhanced CT were evaluated.The degree of contrast enhancement at the enhancing portion within the hemangioma was visually assessed using a five-point scale during each phase. For quantitative analysis, the tumor-muscle signal intensity ratio (SIR), the liver-muscle SIR, and the attenuation value of the tumor and liver parenchyma were calculated. The McNemar test and the Wilcoxon's signed rank test were used to assess the significance of differences in the appearances of hemangiomas and in the visual grade of tumor contrast enhancement between Gd-EOB-DTPA-enhanced MRI and enhanced CT. RESULTS:There was no significant difference between Gd-EOB-DTPA-enhanced MRI and enhanced CT in the presence of peripheral nodular enhancement (85% vs 82%), central nodular enhancement (3% vs 3%), diffuse enhancement (11% vs 16%), or arterioportal shunt (23% vs 34%) during arterial phase, or fill-in enhancement (79% vs 80%) during portal venous phase. Prolonged enhancement during equilibrium phase was observed less frequently on Gd-EOB-DTPA-enhanced MRI than on enhanced CT (52% vs 100%, P < 0.001). On visual inspection, there was significantly less contrast enhancement of the enhancing portion on Gd-EOB-DTPA-enhanced MRI than on enhanced CT during the arterial (3.94 ± 0.98 vs 4.57 ± 0.64, respectively, P < 0.001), portal venous (3.72 ± 0.82 vs 4.36 ± 0.53, respectively, P < 0.001), and equilibrium phases (2.01 ± 0.95 vs 4.04 ± 0.51, respectively, P < 0.001). In the quantitative analysis, the tumor-muscle SIR and the liver-muscle SIR observed with Gd-EOB-DTPA-enhanced MRI were 0.80 ± 0.24 and 1.28 ± 0.33 precontrast, 1.92 ± 0.58 and 1.57 ± 0.55 during the arterial phase, 1.87 ± 0.44 and 1.73 ± 0.39 during the portal venous phase, 1.63 ± 0.41 and 1.78 ± 0.39 during the equilibrium phase, and 1.10 ± 0.43 and 1.92 ± 0.50 during the hepatobiliary phase, respectively. The attenuation values in the tumor and liver parenchyma observed with enhanced CT were 40.60 ± 8.78 and 53.78 ± 7.37 precontrast, 172.66 ± 73.89 and 92.76 ± 17.92 during the arterial phase, 152.76 ± 35.73 and 120.12 ± 18.02 during the portal venous phase, and 108.74 ± 18.70 and 89.04 ± 7.25 during the equilibrium phase, respectively. Hemangiomas demonstrated peak enhancement during the arterial phase, and both the SIR with Gd-EOB-DTPA-enhanced MRI and the attenuation value with enhanced CT decreased with time. The SIR of hemangiomas was lower than that of liver parenchyma during the equilibrium and hepatobiliary phases on Gd-EOB-DTPA-enhanced MRI. However, the attenuation of hemangiomas after contrast injection was higher than that of liver parenchyma during all phases of enhanced CT. CONCLUSION:Prolonged enhancement during the equilibrium phase was observed less frequently on Gd-EOB-DTPA-enhanced MRI than enhanced CT, which may exacerbate differentiating between hemangiomas and malignant tumors.

Epstein-Barr virus-associated gastric carcinoma: Evidence of age-dependence among a Mexican population

AIM： To investigate features of Epstein-Barr virus （EBV）associated gastric carcinoma （EBVaGC） among a Mexican population. METHODS： Cases of primary gastric adenocarcinoma were retrieved from the files of the Departments of Pathology at the Instituto Nacional de Cancerologia and the Instituto Nacional de la Nutrición in Mexico City. The anatomic site of the gastric neoplasia was identified, and carcinomas were histologically classified as intestinal and diffuse types and subclassified as proposed by the Japanese Research Society for Gastric Cancer. EBV-encoded small non-polyadenylated RNA-1 （EBER-1） in situ hybridization was conducted to determine the presence of EBV in neoplastic cells. RESULTS： We studied 330 consecutive, non-selected, primary gastric carcinomas. Among these, there were 173 male and 157 female patients （male/female ratio 1.1/1）. EBER-1 was detected in 24 （7.3%） cases （male/ female ratio： 1.2/1）. The mean age for the entire group was 58.1 years （range： 20-88 years）, whereas the mean age for patients harboring EBER-1-positive gastric carcinomas was 65.3 years （range： 50-84 years）. Age and histological type showed statistically significant differences, when EBER-1-positive and -negative gastric carcinomas were compared. EBER-1 was detected in hyperplastic- and dysplastic-gastric mucosa surrounding two EBER-1-negative carcinomas, respectively. CONCLUSION： Among Latin-American countries, Mexico has the lowest frequency of EBVaGC. Indeed, the Mexican population 〉50 years of age was selectively affected. Ethnic variations are responsible for the epidemiologic behavior of EBVaGC among the worldwide population.

Human papillomavirus in esophageal squamous cell carcinoma in Colombia and Chile

AIM： To examine the presence of human papillomavirus （HPV） in esophageal squamous cell carcinoma （ESCC） specimens collected from Colombia and Chile located in the northern and southern ends of the continent, respectively.METHODS： We examined 47 and 26 formalin-fixed and paraffin-embedded ESCC specimens from Colombia and Chile, respectively. HPV was detected using GP5＋/GP6＋ primer pair for PCR, and confirmed by Southern blot analysis. Sequencing analysis of L1 region fragment was used to identify HPV genotype. In addition, P16^INK4A protein immunostaining of all the specimens was conducted.RESULTS： HPV was detected in 21 ESCC specimens （29%）. Sequencing analysis of L1 region fragment identified HPV-16 genome in 6 Colombian cases （13%） and in 5 Chilean cases （19%）. HPV-18 was detected in i0 cases （21%） in Colombia but not in any Chilean case. Since Chilean ESCC cases had a higher prevalence of HPV-16 （without statistical significance）, but a significantly lower prevalence of HPV-18 than in Colombian cases （P = 0.011） even though the two countries have similar ESCC incidence rates, the frequency of HPV-related ESCC may not be strongly affected by risk factors affecting the incidence of ESCC. HPV-16 genome was more frequently detected in p16 positive carcinomas, although the difference was not statistically significant. HPV-18 detection rate did not show any association with p16 expression. Well-differentiated tumors tended to have either HPV-16 or HPV-18 but the association was not statistically significant. HPV genotypes other than HPV-16 or 18 were not detected in either country.CONCLUSION： HPV-16 and HPV-18 genotypes can be found in ESCC specimens collected from two South American countries. Further studies on the relationship between HPV-16 presence and p16 expression in ESCC would aid understanding of the mechanism underlying the presence of HPV in ESCC.

E-cadherin and beta-catenin expression in Epstein-Barr virus-associated gastric carcinoma and their prognostic significance

AIM： To examine the role of E-cadherin and betacatenin in carcinogenesis and to assess their prognostic implication in Epstein-Barr virus-associated gastric carcinomas （EBV-GCs）. METHODS： We compared the frequency of E-cadherin and beta-catenin expression in 59 EBV-GCs and 120 non-EBV-GCs, and examined the association between patients＇ prognosis and the expressions of these proteins. RESULTS： Neither the cellular-membranous nor the cytoplasmic E-cadherin expression showed any difference between EBV-GCs and non-EBV-GCs. On the other hand, loss of membranous expression of beta- catenin occurred more frequently in non-EBV-GCs than EBV-GCs [odds ratio = 0.41; 950 confidence interval （CI）, 0.19-0.90]. Furthermore, the nuclear and/or cytoplosmic expression of beta-catenin was seen more frequently in EBV-GCs than non-EBV-GCs （odds ratio = 2.23; 95% CI, 0.97-5.09）, and was observed in a larger proportion of carcinoma cells of EBV-GCs than non-EBV-GCs （P = 0.024）. Survival analysis for non-EBV-GC revealed that lymph node metastasis was significantly associated with poor prognosis （P 〈 0.001）. Among EBV- GCs, the depth of invasion （P = 0.005）, lymph node metastasis （P = 0.004） and an intestinal type by Lauren classification （hazard ratio = 9.47; 95% CI, 2.67-33.6） were significantly associated with poor prognosis. On the other hand, nuclear and/or cytoplasmic expression of beta-catenin was associated with a better prognosis in patients with EBV-GC （hazard ratio = 0.32; 95% CI, 0.11-0.93）. CONCLUSION： We observed more frequent preservation of beta-catenin in cell membrane and accumulation in nuclei and/or cytoplasm in EBV-GCs than in non-EBV- GCs. Factors involved in the prognosis of EBV-GCs and non-EBV-GCs are different in the two conditions.

Human papilloma virus and esophageal carcinoma in a Latin-American region

AIM:To investigate the presence of high-risk human papilloma virus (HPV) in esophageal squamous cell carcinomas (ESCCs) in a non-selected Mexican population.METHODS: Cases with a pathological diagnosis of squamous cell carcinoma of the esophagus were obtained from Department of Pathology files, at the National Cancer Institute in Mexico City during the period between 2000 and 2008. Slides from each case were reviewed and cases with sufficient neoplastic tissue were selected for molecular analysis. DNA was extracted from paraffin-embedded tissue samples for polymerase chain reaction analysis to detect HPV DNA sequences. Demographic and clinical data of each patient were retrieved from corresponding clinical records.RESULTS: HPV was detected in 15 (25%) of ESCCs. HPV-16 was the most frequently observed genotype, followed by HPV-18; HPV-59 was also detected in one case. Unfortunately, HPV genotype could not be established in three cases due to lack of material for direct sequencing, although universal primers detected the presence of HPV generic sequences. No low-risk HPV genotypes were found nor was HPV-16/18 co-infection. HPV presence in ESCC was not significantly associated with gender, age, alcohol consumption, smoking, anatomic location, or histologic grade. All patients belonged to low and very low socioeconomic strata, and were diagnosed at advanced disease stage. Male patients were most commonly affected and the male:female ratio in HPV-positive ESCC increased two- fold in comparison with HPV-negative cases (6.5:1 vs 3.1:1).CONCLUSION: High prevalence of high-risk HPV in ESCC in Mexico does not support the hypothesis that HPV-associated ESCC is more common in areas with higher ESCC incidence rates.

Systemic review and network meta-analysis:Prophylactic antibiotic therapy for spontaneous bacterial peritonitis

BACKGROUND Spontaneous bacterial peritonitis(SBP)is an important prognostic factor for outcomes in patients with cirrhosis.Antibiotic prophylaxis is recommended in patients at high risk for developing SBP,but the choice of antibiotics remains unclear.AIM To evaluate the efficacy of various antibiotics for prophylaxis of SBP based on randomized control trials(RCTs).METHODS Electronic databases were searched through November 2018 for RCTs evaluating the efficacy of therapies for primary or secondary prophylaxis of SBP.The primary outcome was the development of SBP.Sensitivity analyses limited to studies of primary or secondary prophylaxis and studies reported after 2010 were performed.The secondary outcome was the risk of all-cause mortality or transplant.The outcomes were assessed by rank of therapies based on network meta-analyses.Individual meta-analyses were also performed.RESULTS Thirteen RCTs(1742 patients)including norfloxacin,ciprofloxacin,rifaximin,trimethoprim-sulfamethoxazole(TMP-SMX),or placebo/no comparator were identified.Individual meta-analyses showed superiority of rifaximin over norfloxacin as well as norfloxacin and TMP-SMX over placebo.Network metaanalysis demonstrated the rank of efficacy in reducing the risk of SBP as:Rifaximin,ciprofloxacin,TMP-SMX,norfloxacin,and placebo/no comparator.Rifaximin ranked highest in sensitivity analyses limited to studies of primary or secondary prophylaxis and studies reported after 2010.Similarly,rifaximin ranked highest in reducing the risk of death/transplant.CONCLUSION The present comprehensive network meta-analysis provides RCT based evidence for superior efficacy of rifaximin compared to other antibiotics for the prophylaxis of SBP and reducing risk of death/transplant.Further RCTs are warranted to confirm our findings.

Rare histological subtype of invasive micropapillary carcinoma in the ampulla of Vater: A case report

BACKGROUND Carcinoma of the ampulla of Vater is an uncommon ampullo-pancreatobiliary neoplasm,and the most common histological type is adenocarcinoma with a tubular growth pattern.Invasive micropapillary carcinoma(IMPC)is an aggressive variant of adenocarcinoma in several organs that is associated with lymph node metastasis and poor prognosis.IMPC was first described as a histological subtype of breast cancer;however,IMPC of the ampulla of Vater is extremely rare,with only three articles reported in the English literature.CASE SUMMARY We have reported a case of IMPC of the ampulla of Vater in an 80-year-old man.Microscopically,the surface area of the carcinoma was composed of tubulopapillary structures mimicking intra-ampullary papillary-tubular neoplasm,and the deep invasive front area exhibited a pattern of IMPC.The carcinoma showed lymphatic invasion and extensive lymph node metastasis.The immunohistochemical study revealed mixed intestinal and gastric/pancreatobiliary phenotypes.CONCLUSION This rare subtype tumor in the ampulla of Vater showed a histologically mixed phenotype and exhibited aggressive behavior.

Serum manganese superoxide dismutase and thioredoxin are potential prognostic markers for hepatitis C virus-related hepatocellular carcinoma

AIM:To evaluate the clinical significance of oxidative stress markers in patients with hepatitis C virus(HCV)related hepatocellular carcinoma(HCC).METHODS:Sixty-four consecutive patients who were admitted to Kagoshima University Medical and Dental Hospital were enrolled in this retrospective study.All patients had chronic liver disease(CLD) due to infection with HCV.Thirty patients with HCV-related HCC,34 with HCV-related CLD without HCC(non-HCC),and 20 healthy volunteers(HVs) were enrolled.Possible associations between serum manganese superoxide dismutase(MnSOD) and thioredoxin(TRX) levels and clinical parameters or patient prognosis were analyzed over a mean follow-up period of 31.7 mo.RESULTS:The serum MnSOD levels were significantly higher in patients with HCV-related HCC than in patients without HCC(P = 0.03) or HVs(P < 0.001).Similarly,serum TRX levels were also significantly higher in patients with HCV-related HCC than in patients without HCC(P = 0.04) or HVs(P < 0.01).However,serum levels of MnSOD and TRX were not correlated in patients with HCC.Among patients with HCC,the overall survival rate(OSR) was lower in patients with MnSOD levels ≥ 110 ng/mL than in patients with levels < 110 ng/mL(P = 0.01),and the OSR tended to be lower in patients with TRX levels < 80 ng/mL(P = 0.05).In addition,patient prognosis with HCC was poorest with serum MnSOD levels ≥ 110 ng/mL and serum TRX levels < 80 ng/mL.Furthermore,a multivariate analysis using a Cox proportional hazard model and serum levels of five factors(MnSOD,prothrombin time,serum albumin,serum α-fetoprotein(AFP),and serum des-γ-carboxy prothrombin) revealed that MnSOD levels ≥ 110 ng/mL(risk ratio:4.12,95% confidential interval:1.22-13.88,P = 0.02) and AFP levels ≥ 40 ng/mL(risk ratio:6.75;95% confidential interval:1.70-26.85,P < 0.01) were independent risk factors associated with a poor patient prognosis.CONCLUSION:Serum MnSOD and TRX levels are potential clinical biomarkers that predict patient prognosis in HCV-related HCC.

Simultaneous operation for cardiac disease and gastrointestinal malignancy

AIM: To investigate the safety of performing simulta-neous cardiac surgery and a resection of a gastrointes-tinal malignancy. METHODS: Among 3664 elective cardiac operations performed in adults at Kagoshima University Hospi-tal from January 1991 to October 2009, this study reviewed the clinical records of the patients who un-derwent concomitant cardiac surgery and a gastroin-testinal resection. Such simultaneous surgeries were performed in 15 patients between January 1991 and October 2009. The cardiac diseases included 8 cases of coronary artery disease and 7 cases with valvular heart disease. Gastrointestinal malignancies included 11 gas-tric and 4 colon cancers. Immediate postoperative andlong-term outcomes were evaluated. RESULTS: Postoperative complications occurred in 5 patients(33.3%), including strokes(n = 1), respiratory failure requiring re-intubation(n = 1), hemorrhage(n = 2), hyperbilirubinemia(n = 1) and aspiration pneu-monia(n = 1). There was 1 hospital death caused by the development of adult respiratory distress syndrome after postoperative surgical bleeding followed aortic valve replacement plus gastrectomy. There was no car-diovascular event in the patients during the follow-up period. The cumulative survival rate for all patients was 69.2% at 5 years. CONCLUSION: Simultaneous procedures are accept-able for the patients who require surgery for both car-diac diseases and gastrointestinal malignancy. In par-ticular, the combination of a standard cardiac operation, such as coronary artery bypass grafting or an isolated valve replacement and simple gastrointestinal resection, such as gastrectomy or colectomy can therefore be safely performed.