DEN Group Research Articles

A Method for Specifying Complete Signature Randomization and an Algebraic Algorithm Based on It

To eliminate the limitations of signature randomization in known algebraic algorithms with a hidden group, the security of which is based on the computational complexity of solving large systems of power equations, a method for ensuring complete randomization is proposed. Based on this method, a new algorithm of the indicated type was developed, using a four-dimensional finite non-commutative associative algebra as an algebraic basis. We obtained estimates of the security of algorithms to direct attacks as well as from attacks based on known signatures, which confirm the effectiveness of the proposed signature randomization method. Due to the relatively small size and signature of the public and private keys, the developed algorithm is of interest as a potential practical post-quantum digital signature scheme.

Early Enteral Nutrition and Clinical Outcomes in COPD Patients Requiring Mechanical Ventilation

Early commencement of enteral nutrition (EEN) in critically ill patients requiring mechanical ventilation may improve outcomes. But there is a lack of enough data regarding EEN effects on COPD exacerbation patients’ outcomes. This retrospective study involved 129 COPD exacerbation patients who received invasive mechanical ventilation in ICU. The clinical outcomes were compared based on the timing of enteral nutrition (<48h vs >48h) during 60 days of ICU stay. We surveyed and analyzed mortality, pleural effusion, ventilator-associated pneumonia, weaning failure, cardiac arrhythmias, GI bleeding, electrolyte imbalances, renal dysfunction and length of ICU stay. All analyses were performed using SPSS software version 22.0. 129 COPD-exacerbated patients (EEN group n=66; DEN group n=63) who met the inclusion criteria were enrolled in the study. EEN group had a lower death rate (39% vs 44.4%) than the DEN group, but no significant difference was found in the overall mortality during the 60-day follow-up (P 0.561). The EEN group also had lower ICU stay and pleural effusion rate than the DEN group (P:0.006 and 0.020 respectively). No significant differences were found in other outcomes. Early enteral nutrition might be associated with shorter ICU stay and lower odd ratio of acquisition of pleural effusion in COPD patients requiring invasive mechanical ventilation. EEN could not decrease mortality rate compared with DEN in the current study.

Belt electrode tetanus muscle stimulation reduces denervation-induced atrophy of rat multiple skeletal muscle groups

Belt electrode-skeletal muscle electrical stimulation (B-SES) involves the use of belt-shaped electrodes to contract multiple muscle groups simultaneously. Twitch contractions have been demonstrated to protect against denervation-induced muscle atrophy in rats, possibly through mitochondrial biosynthesis. This study examined whether inducing tetanus contractions with B-SES suppresses muscle atrophy and identified the underlying molecular mechanisms. We evaluated the effects of acute (60 Hz, 5 min) and chronic (60 Hz, 5 min, every alternate day for one week) B-SES on the tibialis anterior (TA) and gastrocnemius (GAS) muscles in Sprague–Dawley rats using belt electrodes attached to both ankle joints. After acute stimulation, a significant decrease in the glycogen content was observed in the left and right TA and GAS, suggesting that B-SES causes simultaneous contractions in multiple muscle groups. B-SES enhanced p70S6K phosphorylation, an indicator of the mechanistic target of rapamycin complex 1 activity. During chronic stimulations, rats were divided into control (CONT), denervation-induced atrophy (DEN), and DEN + electrically stimulated with B-SES (DEN + ES) groups. After seven days of treatment, the wet weight (n = 8–11 for each group) and muscle fiber cross-sectional area (CSA, n = 6 for each group) of the TA and GAS muscles were reduced in the DEN and DEN + ES groups compared with that in the CON group. The DEN + ES group showed significantly higher muscle weight and CSA than those in the DEN group. Although RNA-seq and pathway analysis suggested that mitochondrial biogenesis is a critical event in this phenomenon, mitochondrial content showed no difference. In contrast, ribosomal RNA 28S and 18S (n = 6) levels in the DEN + ES group were higher than those in the DEN group, even though RNA-seq showed that the ribosome biogenesis pathway was reduced by electrical stimulation. The mRNA levels of the muscle proteolytic molecules atrogin-1 and MuRF1 were significantly higher in DEN than those in CONT. However, they were more suppressed in DEN + ES than those in DEN. In conclusion, tetanic electrical stimulation of both ankles using belt electrodes effectively reduced denervation-induced atrophy in multiple muscle groups. Furthermore, ribosomal biosynthesis plays a vital role in this phenomenon.

Ameliorative Effects of Thunbergia erecta L. Leaves Against the Initiation of Hepatocarcinogenesis Induced by Diethylnitrosamine in the Rat Model

Thunbergia erecta L. contains cytotoxic and liver-protective compounds. Thunbergia erecta L. leaves were macerated in 70% aqueous ethanol, then fractionated with ethyl acetate (9.3 g) and butanol (12.7 g), and attenuated Den-induced liver cancer in a Wistar rat experimental model. Ethyl acetate and butanol fractions were chromatographed using column chromatography and solid-phase extraction (SPE); Vicenin-II (1), kaempferol (2), biochanin A, sissotrin 7-O-β-glucopyranoside (3), gentianose (4), acacetin 7-O-β-glucopyranoside (5), apigenin 7-O-β-glucopyranoside (6), and rosmarinic acid (7) were extracted, and their structures were determined using NMR spectroscopy and ESI-mass spectrometry. Sixty rats were divided into six groups (ten each): control group, Den group, doxorubicin/Den-treated group, butanol fraction/Den-treated group, and isolated acacetin 7-O-β-glucopyranoside/Den-treated group. The liver enzymes and proinflammatory biomarkers were used to estimate the liver function. In addition, liver tissues were collected for analysis of oxidative stress markers, gene expression, and histopathology. There is a significant increase in the levels of liver enzymes, AFP, and TNF-ἁ. This was conveyed by a significant increase of IL-1 and caspase-3, elevation of MDA and reduction of GSH, and suppression of Bcl2 and elevation of Bax expression. All parameters in butanol, ethyl acetate fractions, and isolated acacetin 7-O-β-glucopyranoside (major constituents) of T. erecta L. were significantly improved to values close to those of the control group.

Effect of Dendrobine on Cardiac Dysfunction in Rats with Myocardial Infarction by Regulating Autophagy

Aim: To observe Dendrobine (Den) on rats with post-myocardial infarction cardiac dysfunction and mechanism. Materials: Dividing 27 rats as Sham, Model and Den groups, rats treated with two weeks of drug had their cardiac function and structure measured by ultrasound; their myocardial pathological changes observed by HE and Masson staining and observe apoptosis cell number by TUNEL staining; their serum activities of LDH and CK-MB detected by ELISA; myocardial autophagy protein expressions detected by WB and immunohistochemistry. Results: Model group displayed decreased cardiac function levels, enlarged area of myocardial fibrosis, more serum activities of LDH and CK-MB, increased myocardial tissue structural damage and apoptosis cell number, downregulated LAMP2 expression, and up-regulated expressions of Beclin1, LC3-II/LC3-I rate, and P62. To rat victims of myocardial infarction, Den improved cardiac function, reduced area of myocardial fibrosis, compromised activities of serum LDH and CK-MB, and relieved damage in myocardial structure, decreased apoptosis cell number in myocardial tissue, up-regulated the expressions of Beclin1, LAMP2 and LC3-II, and down-regulated P62 to promote the autophagy in myocardium damaged by myocardial infarction. Conclusion: Den alleviates post-myocardial infarction cardiac dysfunction through improvement of autophagosomes formation and autophagic flux via Beclin1/LAMP2 pathway.

Study on the Mechanism of Improving Mice with Atherosclerosis Using Dendrocrepine

Aim: To discuss Den on apoptosis and Nrf2/ARE in atherosclerotic vulnerable plaque of apolipoprotein in E (ApoE)-/- mice. Methods: Randomly dividing ApoE-/- mice as 5 groups including Normal, Model, Den-L (10 mg/kg), Den-M (20 mg/kg) and Den-H (40 mg/kg) groups. The atherosclerotic vulnerable plaque model was established by high-fat feeding and right common carotid artery catheterization (perivascular carotid collar placement, PCR), and Den was given by difference concentration Den, the pathological changes of right common carotid arery, apoptosis of vascular smooth muscle, Bax, Bcl-2 and Caspase-3 proteins expression using IHC and WB assay, Nrf2, ARE and MDA, 8-OHdG and TAC levels were detected. Results: Model group showed typical pathological changes of vulnerable plaque, the apoptosis cell number, Bax, Caspase-3 and MDA, 8-OHdG significantly increased, the Bcl-2, Nrf2, ARE and TAC levels significantly decreased (P < 0.001, respectively); Compared with model group, the plaque of Den groups were reduced and tended to be stable, the apoptosis cell number, Bax, Caspase3 and MDA, 8-OHdG levels significantly decreased, Bcl-2, Nrf2, ARE and TAC levels significantly increased (P<0.05, respectively). Conclusion: Den improves atherosclerotic vulnerable plaque of ApoE-/- mice, which is related to the inhibition of apoptosis and Nrf2/ARE pathway.

Clonorchis sinensis infection contributes to hepatocellular carcinoma progression in rat.

Clonorchis sinensis (C. sinensis) infection is a risk factor for cholangiocarcinoma. Whether it also contributes to the development of hepatocellular carcinoma (HCC) is still unclear. This study explored the potential relationship between C. sinensis infection and HCC. A total of 110 Sprague-Dawley rats were divided into four treatment groups, the negative control group (NC) received intragastric (i.g.) administration of saline, while the clonorchiasis group (CS) received i.g. administration of 150 C. sinensis metacercariae. The diethylnitrosamine-induced group (DEN) received intraperitoneal (i.p.) administration of DEN. The clonorchiasis DEN-induced group (CSDEN) received i.g. administration of 150 C. sinensis metacercariae followed by i.p. administration of DEN. Hematoxylin and eosin staining, immunohistochemistry, and Masson's trichrome staining were performed for histopathological analysis of the isolated tissues. RNA-seq technology and RT-PCR were employed for gene expression. In the DEN group, 15 rats survived, of which 9 developed liver cirrhosis and 7 developed HCC. In the CSDEN group, all of the 17 surviving rats developed cirrhosis, and 15 showed development of HCC. The incidence of liver cirrhosis and HCC was significantly higher in the CSDEN group than in the DEN group. KEGG pathway analysis of the differentially expressed genes suggested significant upregulation in inflammation-associated pathways. Immunohistochemistry and RT-PCR results showed significant upregulation of hepatic progenitor cell markers (CK19, SOX9, EpCAM) in the CS group compared to the NC group, as well as in the CSDEN group compared to the DEN group. Our study suggests that C. sinensis infection increases risk of HCC in a rat model by stimulating proliferation of hepatic progenitor cells.

Difference in the effects of three nutritional pathways on postoperative rehabilitation in patients with gastric cancer and type 2 diabetes mellitus*

Abstract Objective To explore the difference in the effects of three nutritional pathways on the rehabilitation of patients with gastric cancer and diabetes mellitus after operation. Methods Overall, 120 patients were randomly divided into the partial parenteral nutrition (PPN), early enteral nutrition (EEN), and diabetes mellitus special enteral nutritional emulsion (DEN) groups. The differences in the effects of three nutritional modes were compared. Results (1) On postoperative day four, the total protein level in the EEN and DEN groups was significantly higher than that in the PPN group (P < 0.05). On postoperative day ten, body mass index, lymphocyte count, total protein level, and pre-albumin level in the DEN group were significantly higher than those in the PPN group (P < 0.05). (2) On postoperative day four, there was no significant difference in the fasting blood glucose level between the EEN and DEN groups (P > 0.05), but this level was significantly lower than that in the PPN group (P < 0.05). On postoperative day ten, fasting and postprandial blood glucose levels in the DEN group were significantly lower than those in the PPN group. (3) On postoperative day four, the C-reactive protein level in the DEN group was significantly lower than that in the other groups (P < 0.05). (4) The incidence rates of complications in the PPN, EEN, and DEN groups were 25.0%, 10.0%, and 5.0%, respectively. The incidence of complications in the PPN group was significantly higher than that in the other groups. However, there was no significant difference in perioperative indexes among the three groups (P > 0.05). Conclusion Enteral nutrition is more conducive to the recovery of patients with gastric cancer and type 2 diabetes mellitus after operation; the special enteral nutrition emulsion for diabetes mellitus is more effective than the conventional nutrition solution in stabilizing blood sugar levels and reducing the degree of inflammation.

Early versus delayed enteral nutrition for neonatal hypoxic-ischemic encephalopathy undergoing therapeutic hypothermia: a randomized controlled trial

BackgroundThe practice of therapeutic hypothermia (TH) is widely used for neonatal hypoxic-ischemic encephalopathy (HIE) despite its corresponding feeding strategies are still controversial. This randomized controlled trial (RCT) demonstrated to evaluate the effect of early vs. delayed enteral nutrition on the incidence of feeding intolerance (FI) and other association during TH.MethodsThis single center, parallel-group, and no-blinded RCT was processed in a level III, and academic neonatal intensive care unit. Infants who were diagnosed with HIE and undertaken TH from September 2020 to August 2021 were enrolled. Participants were randomized to receive enteral nutrition either during TH/rewarming (early enteral nutrition, EEN) or after TH (delayed enteral nutrition, DEN) according to a recommend enteral feeding protocol. All data were analyzed using SPSS 26.0 software with a p-value< 0.05 was considered statistically significant.ResultsNinety-two infants were enrolled after randomization, but 12 (13.04%) cases including 3 (3.26%) deaths were excluded from eventually analyzed, who did not initiate or discontinue the intervention. 80 cases (42 and 38 in the EEN and DEN group, respectively) who completed the interventions were eventually analyzed. Besides initial time of enteral feeds, two groups had processed the same feeding method. Total 23 (25.0%) cases developed FI, and no difference of morbidity was found between two groups (23.4% vs 26.7%, p = 0.595; Log Rank, p = 0.803). There was no case died or developed late-onset bloodstream and no difference of the incidence of hypoglycemia or weight gain was found (p > 0.05). The percentage of infants who had not reaching the goal of full enteral feeding volume between the two groups was similar (21.43% vs 23.68%, p = 0.809). The average time of parenteral nutrition, reaching full enteral feeds and hospital stay were shorter in the EEN group compared with the DEN group with significant differences (8.81 ± 1.67 vs 10.61 ± 2.06 days, p < 0.001; 9.91 ± 1.88 vs 12.24 ± 2.50 days, p < 0.001; 12.55 ± 4.57 vs 16.47 ± 5.27 days, p = 0.001 respectively).ConclusionsCompared with delayed enteral nutrition, introduction of early enteral nutrition according to a recommend feeding strategy for neonatal HIE undergoing TH may be feasible and safe.FI is frequent in this high-risk group of infants which should not be ignored during feeding process.Trial registrationThe Chinese Clinical Trial Registry,ChiCTR2000038193, 2020-9-13, https://www.chictr.org.cn.

Effect of Locally Administered Denosumab on Bone Graft Healing in Rabbit Critical-Size Calvarial Defects.

Denosumab is a human monoclonal immunoglobulin G2 antibody developed from the ovarian cells of Chinese hamsters. We aimed to histomorphometrically and radiologically evaluate the effects of xenografts used with local denosumab on the healing of defect sites using rabbit skulls. Two 10-mm diameter critical-size defects were created in 16 rabbits. The defect areas were filled with xenografts and xenograft + 3 mg denosumab in the control and denosumab groups (DEN), respectively. We evaluated new bone, residual graft, soft tissue areas, and bone volume in 4- and 8-week study groups. Histomorphometrically, there were no statistically significant differences between groups at both 4 and 8 weeks regarding residual graft, new bone, and soft tissue area (p > 0.05). The 4-week residual graft control group values were significantly higher than the 8-week values (p < 0.05). The soft tissue area was significantly greater in the 4-week compared with the 8-week DEN group (p < 0.05). The radiologically measured total bone volume was significantly greater in the 8-week specimens than in the 4-week specimens (p < 0.05). In this study, denosumab used locally with bone grafts, showed no direct effect on new and total bone volume.

FRUCTOSE DIET INDUCES A METABOLIC REPROGRAMMING TO ENHANCE TUMOR AGGRESSIVENESS

HCC is one of the main causes of cancer-related death worldwide and has third place in mortality. One of the main risk factors is metabolic-associated fatty liver disease (MAFLD), having hepatic steatosis, and related metabolic disorders. Mexican population has the highest obesity rate in both children and adults, and the consumption of hypercaloric diets has been related to that. Also, Mexico is in the top five countries with a higher fructose-enriched diet consumption and has been proved already the relation between fructose consumption and MAFLD. Likewise, fructose has been related to metabolic rewiring in transformed cells, enhancing aggressiveness and survival. To analyze fructose role on aggressiveness promotion of HCC cells. We used C57Bl/6J mice strain (both sex) with a high Fructose diet (Fru) (33% of fructose in the drinking water, ad libitum ). Fru supplementation started with 15 days-old mice, two days after DEN was injected (10 μg/Kg, i.p), and the treatment was ended 8 months later. The UAM ethics committee approved the protocol. In vitro studies were carried out with the Huh-7 HCC cell line and we evaluated metabolic and biochemical parameters. Tissue samples were analyzed by H&E. We observed that the fructose-enriched diet group mice presented fat accumulation in the hepatocytes and also areas with a greater inflammatory infiltrate (Fru). Mice in the fructose-enriched diet + DEN (Fru/HCC) group showed a marked difference between the tumor area and the surrounding tissue and an increase in the number of bile ducts, indicating liver tissue damage. Also, we analyzed the protein content of some lipogenic enzymes and noticed an increment in fatty acid synthase (Fasn) in Fru and Fru/HCC. Due to that, we analyzed if Fru treatment was inducing metabolic rewiring in transformed cells. We obtained metabolic changes in fructose-treated cells, reducing the glycolytic pathway and the traditional Warburg effect. Then we evaluated if the Huh-7 under a Fru treatment was more dependent on mitochondria or glycolysis ATP generation. We observed a reduction in proliferation under oligomycin treatment vs . 2-DG treatment. At least, we evaluated the pentose phosphate pathway (PPP) under a Fru treatment and obtained a higher glucose-6-phosphate dehydrogenase (G-6-P DH) activity with Fru vs. only glucose (Glc). Also, G-6-P DH had a more efficient activity in the presence of Fru because the time to reach the Vmax is lower vs . Glc. Fructose induces a metabolic rewiring in cancer cells to enhance ATP production, NADPH, and nucleotides to sustain the active lipid synthesis and proliferation. The fructose-enriched diets promote an aberrant lipogenic phenotype enhancing tumor aggressiveness. Conacyt, Froteras de la Ciencia 1320. The authors declare that there is no conflict of interest.

Protective and therapeutic role of nano-curcumin against hepatocarcinogenesis in rat

Curcumin medicinal applications are hindered by its well-known unstable metabolic state and also insufficient absorption and bioavailability.In recent years, nano-based drug delivery system has effectively improved the aqueous solubility and bioavailability of curcumin. The current study was established to evaluate the protective and therapeutic effect of nano-curcumin (Nano-Cur) in hepatocellular carcinoma induced in rats. Fifty white albino rats were divided into 5 groups 10 rat of each. Group I served as negative control while group II received DEN with a daily dosage of (20 mg/kg body weight) orally by gavage during twelve weeks. Group III rats received Nano-Cur (20 mg/kg body weight) three days a week by gavage for eighteen weeks. Group IV rats received DEN as group II and treated with Nano-Cur as group III for six weeks. Group V received Nano-Cur as protector, for six weeks before DEN administration with continuous administration with Nano-Cur till the experiment end. Blood and tissue samples were collected and removed for determination of biochemical, antioxidant, cytokine and apoptotic marker p53 examination at the end of experiment. Based on the findings, DEN group indicated significantly increased serum liver function enzymes ALT, AST, ALP this was accompanied with significant decrease in liver antioxidant enzyme activities of (SOD and CAT) along with significant elevation in lipid peroxidation. DEN significantly elevated IL-6 and reduced apoptotic marker p53. The findings of this research indicated that Nano-cur has a anemaelioration effect against DEN carcinogenic effect.

Association between early enteral nutrition and clinical outcome in patients with severe acute heart failure who require invasive mechanical ventilation.

This study aimed to examine the association between early enteral nutrition (EEN) and clinical outcome in patients with severe acute heart failure (AHF). This retrospective observational study enrolled consecutive patients with AHF who required continuous invasive mechanical ventilation (IMV) for >48 h and were admitted to a single-center cardiac care unit (CCU). The primary outcome was CCU length of stay (LoS). We compared patients who were initiated on EN within 48 h of intubation (EEN group) with those who were initiated on EN after 49 h of intubation (delayed EN [DEN] group). Multivariate logistic regression analysis was performed to determine independent factors for primary and secondary outcomes. CCU LoS and IMV time were categorized using the median. We included 86 patients with AHF (EEN group, n = 56; DEN group, n = 30) who met the inclusion criteria. The median CCU LoS was significantly shorter in the EEN group (10 [8-15] days) than in the DEN group (15 [12-26] days, P = .007). Multivariate analysis indicated that time to EN initiation was an independent factor for CCU LoS (odds ratio [OR], 8.39; 95% confidence interval [CI], 2.18-32.20; P = .002), IMV time (OR, 4.84; 95% CI, 1.37-17.20; P = .015), and incidence of infection (OR, 2.73; 95％ CI, 1.04-7.18; P = .042). EEN (within 48 h of intubation) for patients with severe AHF who require continuous IMV might be associated with reduced CCU LoS, IMV time, and incidence of infection.

Microbiome, fibrosis and tumor networks in a non-alcoholic steatohepatitis model of a choline-deficient high-fat diet using diethylnitrosamine

Background & aimsHepatocellular carcinoma in nonalcoholic steatohepatitis is caused by the complex factors of inflammation, fibrosis and microbiomes. We used network analysis to examine the interrelationships of these factors. MethodsC57Bl/6 mice were categorized into groups: choline-sufficient high-fat (CSHF, n = 8), choline-deficient high-fat (CDHF, n = 9), and CDHF+ diethylnitrosamine (DEN, n = 8). All mice were fed CSHF or CDHF for 20 weeks starting at week 8, and mice in the CDHF + DEN group received one injection of DEN at 3 weeks of age. Bacterial gene was isolated from feces and analyzed using Miseq. ResultsThe CSHF group had less fibrosis than the other groups. Tumors were found in 22.2% and 87.5% of the CDHF group and CDHF + DEN groups, respectively. Gene expression in the liver of Cdkn1a (p21: tumor-suppressor) and c-jun was highest in the CDHF group. Bacteroides, Roseburia, Odoribacter, and Clostridium correlated with fibrosis. Streptococcus and Dorea correlated with inflammation and tumors. Akkermansia and Bilophila were inversely correlated with fibrosis and Bifidobacterium was inversely correlated with tumors. ConclusionsDEN suppressed the overexpression of p21 caused by CDHF. Some bacteria formed a relationship networking associated with their progression and inhibition for tumors and fibrosis.

THE EFFECT OF DIFFERENT LEVELS OF DRY SEASON SUPPLEMENTARY FEEDING ON THE REPRODUCTIVE PERFORMANCE OF YANKASA SHEEP IN A TWICE-YEARLY LAMBING PROGRAMME

Ninety-four pregnant Yankasa ewes were divided into 3 groups and fed;(1) 0, (2) 200 or (3) 400 g/head/day of a concentrate supplement in addition to 6 h grazing daily, from 35.9±2.0 days before, to 98. 122.0 days after lambing in the dry season between 2 Jan vary and 15 May 1984 in Zaria, Northern Nigeria. There were no significant differences (P&gt;0.05) bet den groups in litter size and adjusted birth weight in the initial lambing 35.9±2.0 days into the experiment although the latter showed consistent increase with level of supplementation. Adjusted 90-day Weaning weights increased significantly (P&lt;0.05) with each increase in level of supplementation: 5:5, 7.1 and 8.6kg for groups 1, 2 and 3 respectively. Correspondingly, the percentage of lambs weaned increased (48.6, 56.4 and 90.7%) while ewe mortality declined (12.9, 10.0 and 3.0%) with increasing level of supplementation, The percentage of ewes exhibiting oestrus within 98.1±2.0 days postpartum increased with level of supplementation: 77,8,91.3 and 96.8% for groups 1, 2 and 3 respectively. However, of the ewes showing oestrus, there were no significant differences between groups in post-partum intervals to oestrus and conception. Litter size from matings during the experiment showed non-significant increase with level of supplementation. It is concluded that dry season supplementary feeding is essential for optimum reproductive performance of Yankasa sheep in a twice-yearly lambing programme.

Wistar Ratlarında Dietilnitrozamin Uygulanarak Oluşturulan Hepatoselüler Karsinom Modelinde Silimarin’in Koruyucu Etkisi; Kanser Paternleri ve Sitolojik Varyantlar

In this study, the protective effect of silymarin was investigated with histopathological methods in terms of cancer patterns and cytological variants in the hepatocellular carcinoma model created by applying diethylnitrosamine experimentally in rats. For this purpose, the rats were randomly divided into 5 groups. The rats in the control group were fed pellet feed and drinking water for 20 weeks. Animals in Sham group were administered with propylene glycol dissolved in 0.9% NaCl intraperitoneally 3 times a week for 20 weeks. Animals in DEN group administered DEN at a dose of 50 mg/kg once a week for 20 weeks. The rats in the DEN +Silymarin group were Silymarin intraperitoneally administered for 21 weeks 3 times a week at a dose of 100 mg/kg, starting 1 week prior to the administration of DEN. Silymarin was administered to rats in the group of Silymarin 3 times a week intraperitoneally for 20 weeks at a dose of 100 mg/kg. At the end of the study, systemic necropsy of the animals was performed and liver tissue samples were taken. After routine tissue procedures, sections were examined under the light microscope. We observed that Hepatocellular carcinoma occurred in DEN and DEN + Silymarin groups.

The impact of dental status on perceived ability to eat certain foods and nutrient intakes in older adults: cross-sectional analysis of the UK National Diet and Nutrition Survey 2008\u20132014

BackgroundMany factors determine dietary intake in older adults, including physical health, psychological well-being and socio-economic status. Dental status may also be important. The aim was to examine how dental status impacts perceived ability to eat to certain foods, nutrient intake and nutritional status in UK older adults.MethodsData collected by the National Diet and Nutrition Survey Rolling Programme was analysed. A 4-day food diary assessed dietary intake, while a Computer Assisted Personal Interview collected socio-demographic, health behaviour and oral health information. Participants aged 65 years and over (n = 1053) were categorised into three groups according to their dental status: edentate with dentures (E-DEN, n = 292), dentate with dentures (D-DEN, n = 305) or dentate with no dentures (DEN, n = 456). A total of 515 participants provided a blood sample that was used to assess nutrient concentrations including vitamin B12, vitamin C, ferritin, vitamin B6 (pyridoxal-5-phosphate, PLP), retinol, β-carotene and 25-hydroxyvitamin D (25-OH-D). Multiple regression methods were performed to examine cross-sectional associations between dental status, food selection, nutrient intake and nutritional status.ResultsBoth E-DEN and D-DEN groups, compared with the DEN group, were more likely to report difficulty eating apples, raw carrots, lettuce, nuts, well-cooked steak and crusty bread (P < 0.01). No group differences were observed in perceived ability to eat sliced bread, sliced cooked meats and cheese. The E-DEN group compared with the DEN group had lower mean daily intakes of omega 3 fatty acids (P = 0.006), non-starch polysaccharides (P = 0.001), β-carotene (P = 0.001), folate (P = 0.001), vitamin C (P = 0.008), magnesium (P < 0.001) and potassium (P < 0.001), and had lower plasma vitamin B6 PLP (P = 0.001), vitamin C (P = 0.009) and β-carotene (P = 0.015) concentrations, after adjusting for socio-demographic and health behavioural factors. Compared with the DEN group, the D-DEN group did not have lower nutrient intakes or lower blood nutrient concentrations.ConclusionsWithin this sample of older adults, impaired dental status appears to influence food selection, and intake of important nutrients. Future research should focus on developing dental interventions coupled with dietary counselling to encourage the adoption of healthy eating habits in this high-risk population group.

Inhibitory effect of targeted therapies of different signalling pathways in DEN-induced hepatocarcinogenesis in C57BL/6 mice

Background: We describe the successful DEN-induced hepatocarcinogenesis in C57BL/6 mice and investigate the impact of 2 different waiting periods (10 and 12 months), and targeted therapies. Methods: In 60 male mice C57BL/6 DEN-induced hepatocarcinogenesis was achieved. DEN, 5mg/kg bw, was administered intraperitoneally on their 15th day of life. We divided the 60 mice in two major categories according to the post hepatocarcinogenesis waiting periods (10 and 12 months). We then allocated 6 groups with 10 experimental subjects. Group A (10 months) control HCC group, group B (10 months) with administration of anti-Aurora Kinase (PHA-739358, Danusertib), group C (10 months) with administration of anti-WNT1, group D (12 months) control HCC, group E (12 months) with administration of anti-HDAC (Romidepsin) and group F (12 months) with administration of anti-TOPK (HI-TOPK-32). Group G served as control. All animals were sacrificed on the 10th or 12th month of their life according to the protocol. Liver, lungs and serum were retrieved for analyses. Results: Histopathology analysis showed achievement of HCC genesis in the DEN group and administration of targeted therapies intervened in the development of HCC and the cell signaling. Conclusion: DEN-induced hepatocarcinogenesis proved to develop HCC successfully in mice. Targeted therapies for HCC demonstrated an inhibitory effect in the development of HCC and intervened in the HCC development cell signaling.

PO-58 The influence of different intensity exercise on DEN-induced hepatocellular carcinoma and liver metabonomic study

Objective To study the influence of high internal intensity exercise and long-term moderate intensity endurance exercise on DEN-induced hepatocellular carcinoma (HCC) by metabonomics approach, and compare the anti-tumor effect between the different intensity exercise. &#x0D; Methods From 8 to 26 weeks of age, mice with DEN treatment run on the treadmill at different intensity. NC: injection saline only. DEN: injection DEN only. HIT: treatment DEN, running on the treadmill at 1.5km/h, alternating run 2 minutes and rest 2 minutes for 40 minutes once a day, 5 days a week. ET: treatment DEN, running on the treadmill at o.8 km/h for 40 minutes once a day, 5 days a week. Gas chromatograph time-of-flight mass spectrometry (GC−TOFMS) test liver samples of mice and multi-dimensional statistical methods fond differential metabolites in all group.&#x0D; Results We observed that tumor incidence was similar between DEN and HIT; however, it was signicicantly smaller in ET compared to DEN and HIT. Orthogonal partial least squares analysis (OPLS) model was generated based on identified metabolites and showed clear discrimination from only DEN injection group and endurance exercise group, endurance exercise group and high intensity interval exercise group. Compared to DEN group, the level of galactinol, timonacic, glycine, cholesterol, carbamate and palmitic acid significantly decreased in endurance exercise group. Nevertheless, the level of malonic acid, ornithine and glutamic acid obviously increased in endurance group. &#x0D; Conclusions Long-term endurance exercise inclines the hepatic tumor incidence by regulating the level of liver palmitic acid and glutamic acid metabolism . In addition, high intensity interval exercise does not have the effect to prevent tumor development.

Protein phosphatase 2A activation mechanism contributes to JS-K induced caspase-dependent apoptosis in human hepatocellular carcinoma cells

BackgroundJS-K is a nitric oxide (NO) donor and could generate intracellularly high levels of NO. The study explores PP2A as a tumor suppressor is a major determinant mediating JS-K-caused apoptosis in human hepatocellular carcinoma (HCC) cells.MethodsThe human HCC cell lines (PLC5, Huh-7, Bel-7402, SMMC-7721 and HepG2) were used to assess effects of JS-K on cell viability, apoptosis induction and PP2A activation. Effects of JS-K on cell morphology, mitochondrial membrane potential, apoptosis and NO levels were determined in HCC cells expressing PP2A. Simultaneously, the expression of PP2A family including PP2A-A(α/β), PP2A-B55, PP2A-C(α/β) and the substrates of PP2A, such as β-catenin, c-Myc and p-Bcl-2 (Ser70) were detected in sensitive HCC cells. Furthermore, the role of NO in mediating the expression of PP2A was further validated with Z-VAD-FMK (a caspase inhibitor), Carboxy-PTIO (a NO scavenger), okadaic acid (OA, a PP2A inhibitor) and FTY720 (a PP2A agonist) in JS-K treated cells. In addition, the genetic manuplation of PP2A including overexpression and knockdown have been also performed in JS-K treated cells. Moreover, the rat model of primary hepatic carcinoma was established with diethylnitrosamine for 16 weeks to verify the anti-tumor effects of JS-K in vivo. Immunohistochemical and Western blot analysis were used to determine the expression of proteins in rat primary hepatic carcinoma tissues.ResultsJS-K significantly inhibited cell proliferation, increased apoptosis rate and activated PP2A activity in five HCC cells viability, especially SMMC7721 and HepG2 cells. It was characterized by loss of mitochondrial membrane potential, significant externalization of phosphatidylserine, nuclear morphological changes. Moreover, JS-K enhanced Bax-to-Bcl-2 ratio, released cytochrome c (Cyt c) from mitochondria, activated cleaved-caspase-9/3 and the cleavage of PARP, and decreased the expression of X-linked inhibitor of apoptosis protein (XIAP). Both Z-VAD-FMK and Carboxy-PTIO suppressed the activation of cleaved-caspase-9/3 and of cleaved-PARP in JS-K-treated sensitive HCC cells. Simultaneously, JS-K treatment could lead to the activation of protein phosphatase 2A-C (PP2A-C) but not PP2A-A and PP2A-B55, which subsequently inactivated and dephosphorylated the PP2A substrates including β-catenin, c-Myc, and p-Bcl-2 (Ser70). However, silencing PP2A-C could abolish both the activation of PP2A-C and down-regulation of β-catenin, c-Myc and p-Bcl-2 (Ser70) in sensitive HCC cells. Conversely, PP2A overexpression could enhance the effects of JS-K on activation of PP2A and down-regulation of β-catenin, c-Myc and p-Bcl-2 (Ser70). In addition, adding okadaic acid (OA), a PP2A inhibitor, abolished the effects of JS-K on apoptosis induction, PP2A activation and the substrates of PP2A dephosphorylation; FTY720, a PP2A agonist, enhanced the effects of JS-K including apoptosis induction, PP2A activation and the substrates of PP2A dephosphorylation. The mice exhibited a lower number and smaller tumor nodules in response to JS-K-treated group. A marked increase in the number of hepatocytes with PCNA-positive nuclei (proliferating cells) was evident in DEN group and tended to decrease with JS-K treatment. Furthermore, JS-K treatment could induce PP2A activation and the substrates of PP2A inactivation such as β-catenin, c-Myc and p-Bcl-2(Ser70) in DEN-induced hepatocarcinogenesis.ConclusionsHigh levels of NO released from JS-K induces a caspase-dependent apoptosis through PP2A activation.