Bioinformatics analysis reveals novel core genes associated with nonalcoholic fatty liver disease and nonalcoholic steatohepatitis.

Nonalcoholic fatty liver disease (NAFLD) is the most frequent liver disease and associated with a wide spectrum of hepatic disorders ranging from nonalcoholic fatty liver (NAFL) to nonalcoholic steatohepatitis (NASH), cirrhosis, and hepatocellular carcinoma (HCC). NASH is projected to become the most common indication for liver transplantation, and the annual incidence rate of NASH-related HCC is 5.29 cases per 1000 person-years. Owing to the epidemics of NAFLD and the unclear mechanism of NAFLD progression, it is important to elucidate the underlying NAFLD mechanisms in detail. NASH is mainly caused by the development of NAFL Therefore, it is also of great significance to understand the mechanism of progression from NAFL to NASH. Gene expression chip data for NAFLD and NASH were downloaded from the Gene Expression Omnibus database to identify differentially expressed genes (DEGs) between NAFLD and normal controls (called DEGs for NAFLD), as well as between NASH and normal tissue (called DEGs for NASH-Normal), and between NASH and NAFL tissue (called DEGs for NASH-NAFL). For DEGs for the NAFLD group, key genes were identified by studying the form of intersection. Potential functions of DEGs for NASH were then analyzed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. A protein-protein interaction network (PPI) was constructed using the STRING database. A total of 249 DEGs and one key gene for NAFLD were identified. For NASH-Normal, 514 DEGs and 11 hub genes were identified, three of which were closely related to the survival analysis of HCC, and potentially closely related to progression from NASH to HCC. One key gene for NASH-NAFL (AKR1B10) was identified. These genes appear to mediate the molecular mechanism underlying NAFLD and may be promising biomarkers for the presence of NASH.

Roles of Autophagy and Protein Kinase C-epsilon in Lipid Metabolism of Nonalcoholic Fatty Liver Cell Models.

OBJECTIVE: To investigate interaction between autophagy and PKC-ε in lipid metabolism of NAFLD cell models. METHODS: HL-7702 cells and SK-HEP-1 cells were cultured in vitro as NAFLD cell models and treated with RAPA to induce autophagy. 3-MA was used to inhibit cell autophagy. And HL-7702 and SK-HEP-1 cell were ordinary cultured as control groups. Cell viability was determined by MTT colorimetric assay. The levels of TG, TC and PKC-ε were detected by ELISA. PKC-ε was detected by IF. LC3-II/LC3-I, P62, IRS-1, IRS-2, PI3Kp85, mTOR were detected by Western-blot. SPSS 20 software was used for statistical analysis. RESULTS: The values of PKC-ε were the highest in the steatosis groups (HL-7702 cells were 91.10%, SK-HEP-1 cells were 98.20%). Compared with the steatosis groups, the LC3-II/LC3-I ratio in the induced autophagy groups increased obviously (p <0.05). P62/ß-actin grayscale ratio of the induced autophagy groups decreased significantly compared with the steatosis group (p <0.05). MTOR/ß-actin grayscale ratio in the induced autophagy groups were significantly lower than those in the steatosis groups (p <0.05). PI3Kp85, IRS-1 and IRS-2/ß-actin grayscale ratio of the induced autophagy groups increased significantly compared with the steatosis group (p <0.05). CONCLUSION: Up-regulation of autophagy can promote the elimination of liver fat; while down-regulation can promote lipid accumulation. The expression of PKC-ε is positively related to the degree of hepatic steatosis. PI3K was involved in both autophagy and IR induced by PKC-ε. PKC-ε might participate in hepatocyte autophagy by regulating PI3K.

Evaluation of ascitic soluble human leukocyte antigen-G for distinguishing malignant ascites from benign ascites.

The overexpression of soluble human leukocyte antigen-G is associated with malignant tumours. The purpose of our study was to detect soluble human leukocyte antigen-G concentrations in ascites and to evaluate the value of ascitic soluble human leukocyte antigen-G for the diagnosis of malignant ascites. Enzyme-linked immunosorbent assay was used to detect soluble human leukocyte antigen-G levels in 64 patients with malignant ascites and 30 patients with benign ascites. Receiver operating characteristic curves were used to evaluate the diagnostic efficacy of ascitic soluble human leukocyte antigen-G for the detection of malignant ascites. Ascitic soluble human leukocyte antigen-G levels were significantly higher in the malignant ascites group than in the benign ascites group (20.718 ± 3.215 versus 12.467 ± 3.678 µg/L, t = 7.425, p < 0.001). The area under the receiver operating characteristic curve for ascitic soluble human leukocyte antigen-G was 0.957 (95% confidence interval, 0.872-0.992). At a cut-off value of 19.60 µg/L, the sensitivity and specificity of ascitic soluble human leukocyte antigen-G were 87.5% (95% confidence interval, 71.0%-96.5%) and 100% (95% confidence interval, 88.4%-100%), respectively. With respect to area under the receiver operating characteristic curve, sensitivity and specificity, ascitic carcinoembryonic antigen (0.810, 68.75% and 83.33%, respectively) and carbohydrate antigen 19-9 (0.710, 65.63% and 70%, respectively) significantly differed (all p < 0.05). In malignant ascites that were cytology-negative and biopsy-positive, the rate of positivity for ascitic soluble human leukocyte antigen-G was 75%, which was higher than the corresponding rates for ascitic carcinoembryonic antigen (31.25%) and carbohydrate antigen 19-9 (6.25%; both p < 0.05). In conclusion, The detection of ascitic soluble human leukocyte antigen-G exhibited good performance for diagnosing malignant ascites, and particularly those that were cytology-negative and biopsy-positive.

Incidence of gastroesophageal reflux disease in Uygur and Han Chinese adults in Urumqi.

AIM: To investigate the incidence of gastroesophageal reflux disease (GERD) and its related risk factors in Uygur and Han Chinese adult in Urumqi, China. METHODS: A population-based cross-sectional survey was undertaken in a total of 972 Uygur (684 male and 288 female) aged from 24 to 61 and 1023 Han Chinese (752 male and 271 female) aged from 23 to 63 years. All participants were recruited from the residents who visited hospital for health examination from November 2011 to May 2012. Each participant signed an informed consent and completed a GERD questionnaire (Gerd Q) and a lifestyle-food frequency questionnaire survey. Participants whose Gerd Q score was ≥ 8 and met one of the following requirements would be enrolled into this research: (1) being diagnosed with erosive esophagitis (EE) or Barrett's esophagus (BE) by endoscopy; (2) negative manifestation under endoscopy (non-erosive reflux disease, NERD) with abnormal acid reflux revealed by 24-h esophageal pH monitoring; and (3) suffering from typical heartburn and regurgitation with positive result of proton pump inhibitor test. RESULTS: According to Gerd Q scoring criteria, 340 cases of Uygur and 286 cases of Han Chinese were defined as GERD. GERD incidence in Uygur was significantly higher than in Han Chinese (35% vs 28%, χ(2) = 11.09, P < 0.005), Gerd Q score in Uygur was higher than in Han Chinese (7.85 ± 3.1 vs 7.15 ± 2.9, P < 0.005), and Gerd Q total score in Uygur male was higher than in female (8.15 ± 2.8 vs 6.85 ± 2.5, P < 0.005). According to normalized methods, 304 (31%) cases of Uygur were diagnosed with GERD, including 89 cases of EE, 185 cases of NERD and 30 cases of BE; 256 (25%) cases of Han Chinese were diagnosed with GERD, including 90 cases of EE, 140 cases of NERD and 26 cases of BE. GERD incidence in Uygur was significantly higher than in Han Chinese (31% vs 25%, χ(2) = 9.34, P < 0.005) while the incidences were higher in males of both groups than in females (26% vs 5% in Uygur, χ(2) = 35.95, P < 0.005, and 19.8% vs 5.2% in Han, χ(2) = 5.48, P < 0.025). GERD incidence in Uygur male was higher than in Han Chinese male (26% vs 19.8%, χ(2) = 16.51, P < 0.005), and incidence of NERD in Uygur was higher than in Han Chinese (χ(2) = 10.06, P < 0.005). Occupation (r = 0.623), gender (r = 0.839), smoking (r = 0.322), strong tea (r = 0.658), alcohol drinking (r = 0.696), meat-based diet (mainly meat) (r = 0.676) and body mass index (BMI) (r = 0.567) were linearly correlated with GERD in Uygur (r = 0.833, P = 0.000); while gender (r = 0.957), age (r = 0.016), occupation (r = 0.482), strong tea (r = 1.124), alcohol drinking (r = 0.558), meat diet (r = 0.591) and BMI (r = 0.246) were linearly correlated with GERD in Han Chinese (r = 0.786, P = 0.01). There was no significant difference between Gerd Q scoring and three normalized methods for the diagnosis of GERD. CONCLUSION: GERD is highly prevalent in adult in Urumqi, especially in Uygur. Male, civil servant, smoking, strong tea, alcohol drinking, meat diet and BMI are risk factors correlated to GERD.

Identification and characterization of flavonoids from semen zizyphi spinosae by high-performance liquid chromatography/linear ion trap FTICR hybrid mass spectrometry.

Semen zizyphi spinosae (SZS) has been used to treat insomnia and anxiety for thousands of years. In this paper, a novel high-performance liquid chromatography coupled with the photodiode array detector/linear ion trap-MS(n) (HPLC-PDA/LTQ-MS(n)) method was established to separate and identify flavonoids from the extract of SZS. Separation was performed on an HYPERSIL C(18) column by gradient elution using CH(3)CN/H(2)O-CH(3)COOH as the mobile phase at a flow rate of 0.8 ml/min. UV spectral data, accurate molecular weights, and multi-stage MS/MS fragmentation information were obtained. Electrospray ionization/MS/MS fragmentation patterns were proposed. Nineteen flavonoid glycosides were identified or tentatively characterized based on their retention time, UV spectral data, accurate molecular weights, and mass fragmentation behavior. The method was useful for separation and identification of the flavonoid components from SZS and could be applied to other complex samples, especially for minor constituents.

Expression of c-fos in gastric myenteric plexus and spinal cord of rats with cervical spondylosis.

AIM: To determine the expression of c-fos in gastric myenteric plexus and spinal cord of rats with cervical spondylosis and its clinical significance. METHODS: A cervical spondylosis model was established in rats by destroying the stability of cervical posterior column, and the cord segments C(4-6) and gastric antrum were collected 3, 4 and 5 mo after the operation. Rats with sham operation were used as controls. c-fos neuronal counter-staining was performed with an immunohistochemistry method. Every third sections from C(4-6) segments were drawn. The 10 most labeled c-fos-immunoreactive (Fos-IR) neurons were counted, and the average number was used for statistical analysis. The mean of Fos-IR neurons in myenteric plexus was calculated after counting Fos-IR neurons in 25 ganglia from each antral preparation, and expressed as a mean count per myenteric ganglion. RESULTS: There were a few c-fos-positive neurons in the cervical cord and antrum in the control group. There was an increased c-fos expression in model group 3, 4 and 5 mo after operation, whereas there was no significant increase in c-fos expression in the control group at 3, 4 and 5 mo. More importantly, there was a significant difference in c-fos expression between rats followed up for 3 mo and those for 5 mo in the model group (11.20+/-2.26 vs 27.68+/-4.36, P<0.05, for the cervical cord; and 11.3+/-2.3 vs 29.3+/-4.6, P<0.05, for the gastric antrum). There was no significant difference between rats followed up for 3 mo and those for 4 mo and between rats followed up for 4 mo and those for 5 mo in the model group. CONCLUSION: c-fos expression in gastric myenteric plexus was dramatically associated with that in the spinal cord in rats with cervical spondylosis, suggesting that the gastrointestinal function may be affected by cervical spondylosis. If this hypothesis is confirmed by further studies, functional gastrointestinal diseases such as functional dyspepsia and irritable bowel syndrome could be explained by neurogastroenterology.

Genetic polymorphism analysis of cytochrome P4502C19 in Chinese Uigur and Han populations.

OBJECTIVE: To assess the genotype patterns of cytochrome P4502C19 in the Chinese Uigur and Han populations and compare them with other populations. METHODS: Ninety-six Uigur and 104 Han healthy, unrelated subjects (Uigur: 51 males, 45 females, aged from 23 to 30 years; Han: 53 males, 51 females, aged from 21 to 26 years) were genotyped for CYP2C19 by the PCR-RFLP technique to identify the wild-type (wt) gene and two mutations, CYP2C19m1 in exon5 and CYP2C19m2 in exon 4. RESULTS: In the Uigur, the incidence of the poor metabolizer phenotype was 4.17% (4 of 96 subjects with m1/m1 genotype) and that of the extensive metabolizer was 95.83% (92 of 96 subjects), consisting of 6 (6.25%) homozygous for CYP2C19wt (wt/wt) in both exon 4 and exon 5, 83 (86.46%) heterozygous for CYP2C19wt and CYP2C19m1 and 3 (3.13%) heterozygous for both wt/m1 and wt/m2. None of the Uigur subjects was homozygous for CYP2C19m2 (m2/m2) or heterozygous for wt/m2 or m1/m2. Thus in the Uigur, the frequency of the CYP2C19m1 allele was 0.4818, the CYP2C19m2 allele was 0.0078, and that of the CYP2C19wt was 0.5104. In the Han, the frequency of the CYP2C19m1 allele was 0.3333, the CYP2C19m2 allele was 0.0686, and the frequency of the CYP2C19wt was 0.6058. CYP2C19m1 was the majority of the two mutations found in the Uigur, which was consistent with the situation in the Han population, and the frequency of the CYP2C19m1 allele was approximately 60-fold that of the mutant allele CYP2C19m2. On the other hand, there were significant interethnic differences when the Uigur population was compared with other minorities with regard to the CYP2C19 genotype. CONCLUSIONS: The results indicate that the distribution frequency of the poor and the extensive metabolizers in the Uigur population is significantly lower and higher, respectively (P < 0.025), than in the Han population. There are significant differences between the Uigur and Han populations regarding the CYP2C19 allele frequency, which is consistent with results for the Caucasian population. On the other hand, there are significant interethnic differences when the Uigur population is compared with the Miao and Dai populations.

Effect of a single oral dose of rabeprazole on nocturnal acid breakthrough and nocturnal alkaline amplitude.

AIM: To study the effect of rabeprazole (RAB) on nocturnal acid breakthrough (NAB) and nocturnal alkaline amplitude (NAKA) and to compare it with omeprazole (OME) and pantoprazole (PAN). METHODS: By an open comparative study, forty patients with active peptic ulcer were randomly assigned to receive one of the three PPIs (proton pump inhibitor) with a single oral dose. They were divided into RAB group (10 mg), OME group (20 mg) and PAN group (40 mg). Twenty healthy volunteers were enrolled to the control group (without taking any drug). Intragastric pH monitoring was then performed 1 hour before and 24 hours after the dose was given. RESULTS: No clinically undesirable signs and symptoms possibly attributed to the administration of RAB or OME and PAN were recognizable throughout the study period. All subjects completed the study according to the protocol. All data were processed by a computer using the Student t test or t' test followed by an analysis of covariance. P<0.05 was considered to have statistical significance. The intragastric pH of NAB was significantly higher in RAB group (1.84+/-0.55) than in either OME group (1.15+/-0.31) or PAN group (1.10+/-0.30) (both P<0.01). RAB produced a longer sustaining time (4.65+/-1.22 h) on NAKA than OME (3.22+/-1.89 h) (P<0.05), PAN (3.15+/-1.92 h) (P<0.05), and the sustaining time of NAKA in RAB group was longer than that in the healthy control group (P<0.01) too. In addition, RAB produced a much higher pH on NAKA (6.41+/-0.45) in comparison with PAN (6.01+/-0.92) (P<0.05). CONCLUSION: A single oral dose of 10 mg RAB may increase the pH of NAB and shorten the sustaining time of NAB, and it may increase the pH of NAKA as well as prolong the sustaining time of NAKA.