Detection of miRNA-122 expression in serum exosome in patients with HCV infection and its clinical significance

Relationship between intestinal microbial dysbiosis and primary liver cancer

Objective To evaluate the effect of curcumin on the mammalian target of rapamyein(mTOR)signaling pathway during ischemia-reperfusion(I/R)injury in isolated rat lungs. Methods Sixty-four clean-grade healthy male Sprague-Dawley rats, aged 3-4 months, weighing 250-320 g, were divided into 4 groups(n=16 each)using a random number table method: sham operation group(S group), I/R group, low-dose curcumin group(LC group)and high-dose curcumin group(HC group). The rats only received in vitro perfusion without ischemia in S group.Isolated rat lungs were subjected to 60 min of ischemia followed by 75-min reperfusion to establish the lung I/R injury model in I/R group.Curcumin 5 and 10 μmol/L were added to perfusion fluid from the beginning of reperfusion in LC and HC groups, respectively.Airway resistance(Res), lung compliance, perfusion flow(Flow)and pulmonary venous partial pressure of oxygen(PaO2)were recorded at 10 min of first perfusion(T0)and 15, 45 and 75 min of reperfusion(T1-3). Wet/dry lung weight ratio(W/D ratio)was measured at the end of reperfusion.The morphological structure and ultrastructure of lung tissues were observed by using a light microscope and a transmission electron microscope, respectively.The expression of mTOR, Tau protein, nuclear factor kappa B(NF-κB)and tumor necrosis factor-alpha(TNF-α)mRNA in lung tissues was detected by real-time polymerase chain reaction.The expression of mTOR, phosphorylated Tau protein(pS396 Tau protein), NF-κB and TNF-α protein in lung tissues was determined by Western blot. Results Compared with S group, Res at T1-3 and W/D ratio at T3 were significantly increased, lung compliance, Flow and PaO2 were decreased at T1-3, and the expression of mTOR, NF-κB and TNF-α protein and mRNA, Tau protein mRNA and pS396 Tau protein was up-regulated at T3 in I/R, LC and HC groups(P<0.05). Compared with I/R group, Res at T1-3 and W/D ratio at T3 were significantly decreased, lung compliance, Flow and PaO2 were increased at T1-3, and the expression of mTOR, NF-κB and TNF-α protein and mRNA, Tau protein mRNA and pS396 Tau protein was down-regulated at T3 in LC and HC groups(P<0.05). Compared with LC group, Res at T1-3 and W/D ratio at T3 were significantly decreased, lung compliance, Flow and PaO2 were increased at T1-3, and the expression of mTOR, NF-κB and TNF-α protein and mRNA, Tau protein mRNA and pS396 Tau protein was down-regulated at T3 in HC group(P<0.05). The microscopic examination showed that the injury to lung tissues was significantly attenuated in LC and HC groups as compared with I/R group. Conclusion The mechanism by which curcumin reduces I/R injury in isolated rat lungs is related to inhibiting mTOR signaling pathway. Key words: Curcumin; Lung; Reperfusion injury; Protein kinase C

Ruminal microflora is closely correlated with the ruminant’s diet. However, information regarding the effect of high concentrate diets on rumen microflora in yaks is lacking. In the current study, 24 healthy male yaks were randomly assigned to two groups, each fed with different diets: less concentrate (LC; concentrate: coarse = 40: 60) and high concentrate (HC; concentrate: coarse = 80: 20) diets. Subsequently, a 21-day feeding trial was performed with the yaks, and rumen fluid samples were collected and compared using 16 s rRNA sequencing. The results showed that NH3-N, total VFA, acetate, butyrate, isobutyrate, and isovalerate were significantly higher in the HC group than that in the LC group (p < 0.05), while microbial diversity and richness were significantly lower in the HC group (p < 0.05). Principal coordinate analysis indicated that rumen microflora was significantly different in LC and HC groups (p < 0.05). In the rumen, phyla Firmicutes and Bacteroidota were the most abundant bacteria, with Firmicutes being more abundant, and Bacteroidota being less abundant in the HC group than those found in the LC group. Christensenellaceae_R-7_group and Prevotella are the highest abundant ones at the genus level. The relative abundance of Acetitomaculum, Ruminococcus, and Candidatus_Saccharimonas were significantly higher in the HC group than that in the LC group (p < 0.05), while the relative abundance of Olsenella was significantly lower in the HC group than in the LC group (p < 0.05). Compared to the LC group, the relative abundance of Prevotella, Ruminococcus, and Candidatus_Saccharimonas was significantly higher in the HC group. The relative abundances of Prevotella, Prevotellaceae_UCG-003, Olsenella, Ruminococcus, Acetitomaculum, Candidatus_Saccharimonas, and NK4A214_group were correlated with ruminal fermentation parameters (p < 0.05). Furthermore, PICRUSt 2 estimation indicated that microbial genes associated with valine, leucine, and isoleucine biosynthesis were overexpressed in the rumen microflora of yaks in the HC group (p < 0.05). Conclusively, our results suggest that high concentrate diets affect the microflora composition and fermentation function in yak rumen. The present findings would provide new insights into the health of yaks under high concentrate feeding conditions and serve as a potent reference for the short-term fattening processes of yaks.

Short-term or acute temperature stress affect the immune responses and alters the gut microbiota of broilers, but the influences of long-term temperature stress on stress biomarkers and the intestinal microbiota remains largely unknown. Therefore, we examined the effect of three long-term ambient temperatures (high (HC), medium (MC), and low (LC) temperature groups) on the gene expression of broilers’ heat shock proteins (Hsps) and inflammation – related genes, as well as the caecal microbial composition. The results revealed that Hsp70 and Hsp90 levels in HC group significantly increased, and levels of Hsp70, Hsp90, IL-6, TNF-α, and NFKB1 in LC group were significantly higher than in MC group (p < 0.05). In comparison with the MC group, the proportion of Firmicutes increased in HC and LC groups, while that of Bacteroidetes decreased in LC group at phylum level (p < 0.05). At genus level, the proportion of Escherichia/Shigella, Phascolarctobacterium, Parabacteroides,and Enterococcus increased in HC group; the fraction of Faecalibacterium was higher in LC group; and the percentage of Barnesiella and Alistipes decreased in both HC and LC groups (p < 0.05). Functional analysis based on communities’ phylogenetic investigation revealed that the pathways involved in environmental information processing and metabolism were enriched in the HC group. Those involved in cellular processes and signaling, metabolism, and gene regulation were enriched in LC group. Hence, we conclude that the long-term temperature stress can greatly alter the intestinal microbial communities in broilers and may further affect the host’s immunity and health.

We investigated the effect of sodium butyrate feeding on the disruption of ruminal epithelium barrier function in goats fed a high-concentrate diet. A total of 18 male Boer goats (live weight of 31.75 ± 1.35 kg, aged 1 year) were randomly assigned to three groups, which were fed a low-concentrate diet (LC), a high-concentrate diet (HC), or a high-concentrate diet with 1% sodium butyrate by weight (SH) for 9 weeks. We found that the pH of rumen fluid in the SH and LC groups was higher than that in the HC group. The activity of protein kinase C (PKC) kinase in the rumen epithelium was higher in the HC group than that in the LC and SH groups. The mRNA expression and phosphorylated protein levels of mitogen-activated protein kinases (MAPKs) in the rumen epithelium were lower in the SH and LC groups than those in the HC group. The DNA methylation rate of occludin was higher in the HC group than that in the SH and LC groups. The mRNA and protein expression of claudin-1, claudin-4, occludin, and zona occludin-1 was greater in the SH and LC groups than that in the HC group. In addition, sodium butyrate mitigated damage to the rumen epithelium caused by the HC diet. Together, our results suggest that the supply of sodium butyrate reverses the damage of rumen epithelium tight junction by inhibiting PKC and MAPK signaling pathways and is protective to the rumen epithelium during subacute rumen acidosis.

One possible complication of stellate ganglion block (SGB) is respiratory compromise. No study has yet addressed the comparison of its effect on lung function and clinical features, including Horner's syndrome, changes in temperature, sensory and motor functions, and adverse events of lower versus higher concentrations (LC and HC, respectively) of local anesthetics in an ultrasound-guided SGB. Fifty patients were randomized into 1 of 2 groups: the LC group (5 mL of 0.5% mepivacaine) and the HC group (5 mL of 1% mepivacaine). One anesthesiologist performed a C6- SGB under ultrasound guidance. Our primary objective was to compare LC and HC of a local anesthetic in terms of its effect on lung function, and the secondary objective was to compare the clinical features between LC and HC of a local anesthetic. Lung function was compared between the 2 groups using the Mann-Whitney U test. The forced vital capacity at 20 minutes post-SGB was not significantly different between the HC and the LC groups (P = .360); the median difference (95% confidence intervals [CI]) was 1 (-1 to 8). Other parameters of lung function were comparable with the forced vital capacity. Patients in the HC group had significantly greater sensory changes than those in the LC group (% decrease compared with the unblocked side); 95.4 ± 2.1 (CI: 91.11-99.73) vs 87.3 ± 3.5 (CI: 80.12-94.49). Lung function between the LC and HC groups after SGB did not differ significantly. Clinical features between the 2 groups also did not differ clinically, except that patients in the HC group had significantly greater sensory loss in the C6 dermatomes.

Objective To explore the levels of serum hepatitis B surface antigen (HBsAg) in different clinical stages of hepatitis B virus(HBV) infection and their correlation with serum HBV DNA. Methods Five hundred and seventy-five patients with HBsAg-positive and without antiviral therapy were enrolled in this study. The patients were classified into six group: IT group (immune tolerant phase, 120 patients), EPH group (hepatitis B e antigen positive hepatitis, 110 patients), LR group (low replicative phase, 90 patients), ENH group (hepatitis B e antigen negative hepatitis, 110 patients), LC group (liver cirrhosis stage, 85 patients) and HCC group (hepatocellular carcinoma, 60 patients). Serum HBsAg and HBV DNA levels were quantified and analyzed through statistics. Results The levels of serum HBsAg in IT group, EPH group, LR group, ENH group, LC group and HCC group were (4.58 ± 0.40), (4.12 ± 0.50), (2.60 ± 0.68), (3.31 ± 0.27), (2.82 ± 0.57) and (3.03 ± 0.39) lg U/ml respectively, and there was significantly different among the different phase (P<0.01). The levels of serum HBsAg in IT group, EPH group, LR group, ENH group, LC group and HCC group had positive correlation with serum HBV DNA level, and the correlation coefficients were 0.627, 0.579, 0.134, 0.317, 0.159 and 0.224 respectively. Conclusions The levels of serum HBsAg is significantly different in the different clinical phase of HBV infection, and the tendency of the correlation between serum HBsAg and serum HBV DNA gradually weakens in general. Key words: Hepatitis B virus; Hepatitis B surface antigen; Hepatitis B, chronic; Natural history

Background: Salinity is one of the main influencing factors in the culture environment and is extremely important for the survival, growth, development and reproduction of aquatic animals. Methods: In this study, a comparative transcriptome analysis (maintained for 45 days in three different salinities, 30 psu (HC group), 18 psu (MC group) and 3 psu (LC group)) was performed by high-throughput sequencing of economically cultured Penaeus monodon. P. monodon gill tissues from each treatment were collected for RNA-seq analysis to identify potential genes and pathways in response to low salinity stress. Results: A total of 64,475 unigenes were annotated in this study. There were 1,140 upregulated genes and 1,531 downregulated genes observed in the LC vs. HC group and 1,000 upregulated genes and 1,062 downregulated genes observed in the MC vs. HC group. In the LC vs. HC group, 583 DEGs significantly mapped to 37 signaling pathways, such as the NOD-like receptor signaling pathway, Toll-like receptor signaling pathway, and PI3K-Akt signaling pathway; in the MC vs. HC group, 444 DEGs significantly mapped to 28 signaling pathways, such as the MAPK signaling pathway, Hippo signaling pathway and calcium signaling pathway. These pathways were significantly associated mainly with signal transduction, immunity and metabolism. Conclusions: These results suggest that low salinity stress may affect regulatory mechanisms such as metabolism, immunity, and signal transduction in addition to osmolarity in P. monodon. The greater the difference in salinity, the more significant the difference in genes. This study provides some guidance for understanding the low-salt domestication culture of P. monodon.

The aim of the study was to evaluate the impact of carbohydrate-to-fat ratio on body weight and appetite regulation in Wistar rats. Twenty-four Wistar rats were randomized to three dietary groups (n = 8): normal carbohydrate diet (NC), low-carbohydrate diet (LC) and high-carbohydrate diet (HC) for 12 weeks. Body weight and food intake were recorded. Circulating leptin and insulin levels were measured by radioimmunoassay method. The expression levels of leptin receptor, insulin receptor, orexin, neuropeptide Y (NPY), agouti-related protein (AgRP) and melanocortin-4 receptor (MC-4R) in the hypothalamus were also measured by realtime polymerase chain reaction (PCR). In the LC group, food intake reduced while body weight increased significantly compared with the NC and HC groups. Plasma leptin levels increased in the LC (18.5 +/- 8.2 ng/mL) group compared with the NC (8.6 +/- 3.8 ng/mL, P < 0.001) and HC (6.6 +/- 1.9 ng/mL, P < 0.001) groups. Realtime reverse transcription-PCR revealed a decrease in the hypothalamic expression level of only leptin receptor in the LC (0.764, 0.471-4.648 copy/mL) and HC (0.357, 0.129-0.781 copy/mL) groups compared with the NC (1.323, 0.616-2.392 copy/mL; P = 0.01) group, and that there was no significant change in those of insulin receptor, AgRP, Orexin, NPY and MC-4R. Low-carbohydrate, high-fat diet raised body weight, which led to a rising of circulating leptin levels and a reduced expression of leptin receptor in the hypothalamus.

The aim of this study was to examine the effects of supplementing Guizhou black goats with Houttuynia cordata extract (HCE) during summer on growth performance, anti-inflammatory activity, and rumen fermentation parameters. A completely randomized single-factor experimental design was employed. Twenty-four healthy Guizhou black goats, with similar body weights (16.03 ± 0.79 kg), were randomly divided into three groups, with eight replicates per group and one goat per replicate. The control group (CON) was fed a basal diet, the LC and HC groups received the basal diet supplemented with 500 mg/kg and 1,000 mg/kg of HCE, respectively. The ADG in the HC group was significantly higher (p < 0.05) than that in the CON group. The digestibility of DM in both the LC and HC groups was significantly higher (p < 0.05) than in the CON group. Additionally, the digestibility of CP, GE, and NDF in the HC group was significantly higher (p < 0.05) than in the CON group. GSH-Px levels in both the LC and HC groups were significantly higher (p < 0.05) than in the CON group. T-AOC in the HC group was also significantly higher (p < 0.05) than in the CON group. MDA levels in the HC group were significantly lower (p < 0.05) than in both the LC and CON groups (p < 0.05). The TP content in the HC group was significantly higher (p < 0.05) than in the CON group. IgA levels in both the LC and HC groups were significantly higher (p < 0.05) than in the CON group. The levels of IL-6 and IL-8 in the HC group were significantly lower (p < 0.05) than in the LC and CON groups. Interestingly, the IL-10 level in the HC group was significantly higher (p < 0.05) than in the LC and CON groups. The TNF-α level in the HC group was significantly lower (p < 0.05) than in the CON group. The HCE had no significant (p > 0.05) effect on rumen pH, NH3-N, and VFAs. In conclusion, a high dose of HCE improved growth performance, apparent nutrient digestibility, and enhanced antioxidant, immune, and anti-inflammatory responses in goats.

The moon jellyfish (Aurelia aurita) is a scyphozoan frequently maintained in public and private aquaria. Little research has been conducted to investigate the effects of various drugs, such as anesthetics, in this species. Tricaine methanesulfonate (MS-222), a common immersion anesthetic for fish and amphibians, was evaluated in a managed population of moon jellyfish. Twenty-four clinically healthy jellyfish were assigned into three groups of eight for trials of 0.3 g/L MS-222 (low concentration [LC]), 0.6 g/L MS-222 (high concentration [HC]), and a saltwater control. The goal was to evaluate the effects of MS-222 administration on moon jellyfish movement and response to stimuli. Movement and response to stimuli were measured via rocking and probe stimulus tests and observations of bell contraction quality and body tone. These tests were performed at baseline and throughout both drug exposure and recovery periods. A threshold drug effect was defined based on systematic scoring criteria. Additionally, elastomer tags were administered to four of eight animals in each MS-222 group to evaluate response to tag placement after drug exposure. Threshold drug effect was achieved in six of eight individuals in the LC group and eight of eight individuals in the HC group. The LC group had median threshold and recovery times of 12.2 and 10.1 min, respectively, while the HC group had median threshold and recovery times of 4.0 and 19.9 min, respectively. The HC group had significantly faster time to threshold drug effect (P < 0.001) and longer recovery times (P= 0.005) than the LC group. In both the LC and HC tagged group, three of four jellyfish had no reaction to tag placement. All animals recovered uneventfully, and there were no mortalities. MS-222 at 0.3 and 0.6 g/L decreased movement and response to stimuli in moon jellyfish.

Objective To investigate the expression and clinical significance of serum interferon-stimulated gene 20kDa protein (ISG20) of patients with hepatitis B virus infection. Methods Total of 67 patients with chronic hepatitis B (CHB group), 58 patients with liver cirrhosis (LC group), 54 patients with liver cancer (HCC group) and 47 healthy controls were selected in Hanzhong Central Hospital from March 2017 to June 2018. Serum ISG20 protein levels were detected by enzyme-linked immunosorbent assay. The difference of serum ISG levels in different groups were compared. The serum ISG20 level and age, gender, count of red blood cell, count of white blood cell, platelet count and aspartate aminotransferase, alanine aminotransferase, total bilirubin (TBil), direct bilirubin (DBil), albumin, prothrombin, alpha-fetoprotein (AFP) and HBV DNA load of different groups were analyzed by Spearman’s correlation analysis. Results The serum ISG20 level in control group was 9.4 (0.82-25.72) ng/ml, which was significantly lower than that of the CHB group [18.6 (1.87-56.32) ng/ml; Z =-1.567, P = 0.034] and the HCC group [28.2 (3.09-81.42) ng/ml, Z =-1.854, P = 0.021]. The serum ISG-20 level in HCC group was significantly higher than that of CHB group (Z =-1.431, P = 0.041) and LC group [12.9 (2.81-77.54 ) ng/ml; Z =-1.987, P = 0.029). There was significant difference of serum ISG20 level between patients with different Child-Pugh grades in HCC group (H = 6.976, P = 0.031). Serum ISG20 levels of patients in HCC group were positively correlated with AST (r = 0.323, P < 0.001), ALT (r = 0.248, P = 0.036), TBil (r = 0.221, P = 0.031), DBil (r = 0.215, P = 0.043) and AFP (r = 0.176, P = 0.044); but negatively correlated with albumin (r =-0.239, P = 0.019). Conclusions Serum ISG20 levels were elevated in patients with HBV infection, especially in patients with HCC caused by HBV infection. Serum ISG20 levels had a certain correlation with progression and clinical parameters of HBV infection diseases. Key words: Interferon-stimulated gene 20 kDa protein; Hepatitis B virus; Chronic hepatitis B; Liver fibrosis; Hepatocellular carcinoma

This study investigates the effect of the Pain Sensitivity Questionnaire (PSQ) in guiding patient controlled intravenous analgesia (PCIA) on postoperative analgesia in women undergoing cesarean section. A total of 160 women who were to undergo a cesarean section under combined spinal and epidural anaesthesia were included in this study. Women with a preoperative PSQ <4 were randomly divided into a low pain-sensitive control group (LC group), and a low pain-sensitive observation group (LO group), and women with preoperative PSQ >6 were randomly divided into a high pain-sensitive control group (HC group) and a high pain-sensitive observation group (HO group). After the surgery, patients received the pump butorphanol concentration was 3.5 µg·kg-1·h-1 in the LC and HC groups, 3.0 µg·kg-1·h-1 in the LO group and 4.0 µg·kg-1·h-1 in the HO group.To compare the analgesic effects of postoperative PCIA and postoperative recovery in women. Wound pain and uterine contraction pain VAS scores at rest and activity were significantly lower in the LC group than in the LO group at 4 and 8 h postoperatively (P<0.05). Similarly, wound pain and uterine contraction pain VAS scores at rest and activity were significantly lower in the HO group than in the HC group at 8, 12, and 24 h postoperatively (P<0.05). The Ramsay scores were significantly higher in the LC than in the LO groups at 4, 8, 12, 24, and 48 h postoperatively (P<0.05), but there was no statistically significant difference between the Ramsay scores in the HC group and the HO group. There was no statistical difference in any of the post-operative recoveries (P>0.05). Compared to the weight-based postoperative PCIA, the PSQ-based postoperative PCIA has better analgesic effects and can improve maternal satisfaction with postoperative analgesia.

BACKGROUND/OBJECTIVESCalcium is important but deficient in diets of young adult women. This study aimed to examine if cognitive factors and eating behaviors differ according to calcium intake based on the Social Cognitive Theory.SUBJECTS/METHODSSubjects were female college students in Seoul, Korea. Three hundred students completed the questionnaire regarding calcium intake, nutrition knowledge, outcome expectations, self-efficacy and eating behaviors. Data on 240 students were analyzed using t-test or χ2-test. Subjects were categorized into two groups, high calcium intake (HC, ≥ 650 mg/day) and low calcium intake (LC, < 650 mg/day), according to recommended intakes of calcium for women aged 19-29 years.RESULTSThe LC group constituted 77.9% of total subjects. Nutrition knowledge was not different according to calcium intake. Three out of 12 outcome expectations items were significantly different between the HC and LC groups. Subjects in the HC group agreed more strongly with the practical benefits of consuming calcium-rich foods, including 'taste' (P < 0.01) and 'going well with other snacks' (P < 0.05), compared to those in the LC group. Negative expectations of 'indigestion' were stronger in the LC group than HC group (P < 0.001). Among self-efficacy items, perceived ability of 'eating dairy foods for snacks' (P < 0.001), 'eating dairy foods every day' (P < 0.01), and 'eating calcium-rich side dishes at meals' (P < 0.05) differed significantly between the HC and LC groups. Eating behaviors including more frequent consumption of dairy foods, fruits or fruit juice (P < 0.001), anchovy, seaweeds, green vegetables, protein-rich foods (P < 0.05), and less frequent consumption of sweets or soft drinks (P < 0.01) were significantly related to calcium intake.CONCLUSIONSThis study found that outcome expectations, self-efficacy in consuming calcium-rich foods, and eating behaviors are important in explaining calcium intake. Nutrition education needs to address practical benefits, reduce negative expectations of calcium-rich foods, increase self-efficacy, and modify eating behaviors contributing to calcium intake.

BackgroundTumor-associated macrophages (TAMs), inflammatory cells in tumor microenvironment, are crucial for the tumor occurrence and progression which in turn increase the expression of soluble CD163 (sCD163). Nevertheless, not much has been established regarding sCD163 and its connection to HCC diagnosis and prognosis. This study was conducted to evaluate the diagnostic and prognostic role of sCD163 in patients with HCC on top of HCV-related liver cirrhosis. Forty adult patients with HCV-related liver cirrhosis and HCC (HCC group) were randomly selected and subjected to locoregional therapies, either transarterial chemoembolization (TACE) or radiofrequency ablation (RFA). Four patients were excluded because of portal vein invasion. Another group of 20 patients with liver cirrhosis only served as controls (LC group). Routine laboratory studies and abdominal ultrasound were done for all. Alpha-fetoprotein (AFP) and sCD163 were measured twice, at baseline and 1-month post-intervention, using a commercially available enzyme-linked immunosorbent assay kit.ResultsAt baseline, sCD163 showed an insignificant higher value in HCC group (p > 0.05). The best cutoff value for sCD163 and AFP was 6.2 mg/L and 195 ng/mL, respectively. AFP had a larger area under the curve (0.88 vs. 0.767). An overall significant decline was seen in sCD163 after treatment (6.5±1.5 to 3.1±2.5 mg/L; p < 0.001), while AFP showed an insignificant decrease (p > 0.05). Also, sCD163 decreased significantly in the eradicated cases (6.1±1.4 mg/L before intervention vs. 2.3±1.4 mg/L after intervention, p < 0.01), while there was a significant increase in the recurrent cases (8.4±0.4 mg/L before intervention vs. 10.3±1.6 after intervention; p < 0.05). Moreover, sCD163 showed a significant difference in its pre-intervention and post-intervention values between recurrent and eradicated HCC cases (p < 0.01).ConclusionsIt is concluded that sCD163 has a minor role as a diagnostic marker for HCC, yet it could be used as a good prognostic marker in predicting the tumor response to locoregional therapies.