Liver Tissue Concentrations Research Articles

Reactive oxygen species/c-Jun N-terminal kinase/nuclear factor kappa-B signaling molecules are involved in pe-riodontitis-induced liver injury by regulating apoptosis.

The occurrence and development of periodontitis and nonalcoholic fatty liver disease (NAFLD) are closely related to the accumulation of reactive oxygen species (ROS). ROS are involved in regulating the activation of c-Jun N-terminal kinase (JNK)/nuclear factor kappa-B (NF-κB) signaling molecules. When the signaling molecules are overactivated by ROS, the internal environment of the body can be disturbed. Therefore, this study aimed to explore the mechanism by which ROS/JNK/NF-κB signaling molecules are involved in periodontitis-induced liver injury. Twelve SPF male Wistar rats were randomly divided into control and periodontitis groups. The perio-dontitis model of rats was established by wire ligation in the neck of bilateral maxillary first molars. After 8 weeks, the periodontal clinical indexes of the rats were examined, and the rats were sacrificed. Micro-CT reconstruction of a three-dimensional alveolar bone structure and analysis of alveolar bone absorption were conducted. Pathological changes in the periodontal and liver tissues were analyzed by histopathology. MitoSOX red reagent was used to detect the ROS content in liver tissue. Biochemical kits were used to detect liver function and oxidative stress biomarkers. The mRNA expression levels ofinterleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), NF-κB, BCL2-associated X (Bax), and B-cell lymphoma-2 (Bcl-2) in liver tissue were detected through quantitative real-time polymerase chain reaction (qRT-PCR). The protein expression levels of phosphorylated c-Jun N-terminal kinase (P-JNK), JNK, NF-κB, Caspase-3, Bax, and Bcl-2 in liver tissue were detected by Western blot. Apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Micro-CT results showed that the mice in the periodontitis group had obvious alveolar bone resorption and significantly greater distance from the cemento-enamel junction to the alveolar bone crest than those in the control group. Histopathological results showed that a large number of inflammatory cells were infiltrated in the periodontal tissue of the periodontitis group. In addition, the resorption of alveolar ridge bone was obvious and liver tissue structure was destroyed, with balloon-like changes and red lipid droplets. MitoSOX red staining results showed that the ROS level was significantly higher in the liver tissue of the periodontitis group than in that of the control group. Biochemical test results showed that the aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels in the serum of the periodontitis group were higher than those in the serum of the control group. The levels of superoxide dismutase (SOD) and glutathione (GSH) in liver tissue decreased, whereas the that of malondialdehyde (MDA) increased. Western blot and qRT-PCR results revealed that the mRNA levels of IL-6, TNF-α, Bax, and NF-κB and the protein levels of P-JNK/JNK, NF-κB, Caspase-3, and Bax were significantly higher in the liver tissue of the perio-dontitis group than in that of the control group. Meanwhile, the mRNA and protein levels of Bcl-2 were lower in the periodontitis group than in the control group. TUNEL staining showed that the number of apoptotic cells was significantly higher in the periodontitis group than in the control group. ROS/JNK/NF-κB signaling molecules are involved in periodontitis-induced liver injury by regulating apoptosis.

Alteration of Wnt3a overexpression and its early monitoring value during hepatocellular carcinogenesis

Objective: To investigate the dynamic expression pattern of carcinoembryonic Wnt3a and its early monitoring value using a hepatocellular carcinoma model. Methods: Forty-eight Sprague Dawley (SD) rats were fed with pellet feed containing 2-acetylaminofluorene (2-AAF, 0.05%) to induce hepatocarcinogenesis, and control rats were fed a pellet diet. Liver tissue and blood samples were collected every two weeks. Liver tissues were pathologically examined using HE staining and grouped. The gene and Wnt3a mRNA expression were analyzed by genome-wide microarray. The expression and distribution of Wnt3a in liver tissue were analyzed by immunohistochemistry. Wnt3a concentration in liver tissue and serum was quantified by enzyme-linked immunosorbent assay. Statistical methods such as χ2 test, Mann-Whitney test and analysis of variance were used to analyze the differences between groups. Results: According to the pathological examination results, the rat livers were divided into four groups: control, hepatocyte degeneration, precancerous lesions and hepatocellular carcinoma. Genome-wide expression profiling analysis and comparison with the control group revealed that 268 and 312 genes were up-regulated and 57 and 201 genes were down-regulated in the precancerous and cancerous group when signal logarithm ratio (SLR) was >8 log2cy5/cy3, and these significantly altered genes mainly involved in cell proliferation, signal transduction, tumor metastasis, and apoptosis. The expression of Wnt3a at mRNA level was significantly increased in all stages of cancer induction, including degeneration group (1.15±0.24, q=8.227), precancerous group (1.85±0.18, q=12.361) and cancerous group (2.59±0.55, q=18.082). Compared with the control group (0.25±0.11, F=121.103, P<0.001), the degeneration group, the precancerous group and the liver cancer group were up-regulated by 4.6, 7.4 and 10.4-folds, respectively. Immunohistochemistry showed that compared with the control group, the positive rate of Wnt3a in the degeneration group was 66.7% (12/18, χ2=10.701, P=0.001), and both the precancerous and liver cancer groups were positive (9/9, χ2=17.115, P<0.001). Wnt3a expression was gradually increased in liver and blood samples during the process of carcinogenesis, and the difference between two groups was statistically significant (F=176.711, P<0.001). Wnt3a overexpression was secreted into blood stream via cancerous liver tissue, and there was a linear correlation between Wnt3a levels in blood and liver samples (r=0.732, P<0.001). Conclusions: Wnt3a overexpression is closely related with hepatocellular carcinogenesis, and thus may become a new monitoring marker.

Bioaccumulation of Mercury and Radiocesium in Waterfowl Introduced to a Site with Legacy Contamination.

Despite the propensity of waterfowl species to readily accumulate anthropogenic contaminants within polluted environments, few studies have examined bioaccumulation rates over time when entering such a contaminated site. We examined mercury (Hg) and radiocesium (137 Cs) bioaccumulation over time in two waterfowl species released into a wetland system containing legacy contamination on the US Department of Energy's Savannah River Site in South Carolina. Released birds were collected at select time intervals over an exposure period of 94 days. We quantified total Hg concentrations in blood, muscle, and liver tissues, and 137 Cs activity in whole-body and muscle tissues. The relationship between the contaminant burdens of different body tissue types was examined over time. Likely a result of microhabitat selection, mallards in our study readily accumulated both Hg and 137 Cs at consistent rates over time within our study system, while ring-neck ducks did not. The findings demonstrated that whole blood can be used as a robust, nondestructive sampling alternative to estimate Hg burdens within muscle and liver, and whole-body 137 Cs activity is a good predictor of muscle burdens. Understanding such bioaccumulation information in waterfowl is useful for the assessment of the potential health risk in wildlife, as well as being important for human risk assessment toward the consumption of popular game species. Environ Toxicol Chem 2022;41:2479-2487. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Binaprofen induces zebrafish liver injury via the mitochondrial pathway.

Binaprofen (C18H23NO5) is a drug not commercially available that causes liver injury; however, the underlying mechanism is unknown. The aim of the present study was to determine the mechanism underlying binaprofen-induced liver injury at the genetic level. Zebrafish were treated with binaprofen. Serum biomarkers [alanine transaminase (ALT), aspartate transaminase (AST) and lactate dehydrogenase (LDH)], malondialdehyde (MDA) and glutathione (GSH) content analysis, liver cell morphology examination, DAPI staining, electron microscopy, microarray analysis and reverse transcription-quantitative (RT-q)PCR were performed 12, 24 and 48 h post-treatment to analyze the mechanism underlying binaprofen-induced liver injury. Following exposure to binaprofen, zebrafish serum levels of ALT, AST and LDH increased; MDA content of liver tissue increased and GSH content decreased. Liver cells exhibited mild to moderate vacuolization and mitochondria exhibited vacuolization and disrupted cristae. Liver cell apoptosis rate increased. There were 190 common differentially expressed genes at 12, 24 and 48 h. Gene Ontology analysis showed that the function of downregulated genes was primarily associated with ‘DNA replication’, ‘DNA metabolic process’, ‘cell cycle’, ‘cell redox homeostasis’, ‘mitochondrion’ and ‘lipid transport’. The function of upregulated genes was primarily associated with ‘peroxisome proliferator’, ‘oxidation activity’, ‘peroxisome’ and ‘apoptosis’. Pathway analysis showed that downregulated genes were those pertaining to ‘cell cycle’, ‘DNA replication’, ‘ribosome’, ‘spliceosome’, ‘pyrimidine metabolism’, ‘purine metabolism’, upregulated genes were those pertaining to ‘PPAR signaling pathway’, ‘p53 signaling pathway’; RT-qPCR assay supported the microarray results. The mechanism underlying binaprofen-induced liver injury was associated with lipid peroxidation and apoptosis. Binaprofen downregulated genes associated with lipid transport and anti-apoptosis genes, upregulated pro-apoptosis genes and induces liver cell injury via the mitochondrial signaling pathway.

Nicotine poisoning-related deaths in Seoul Capital Area, Korea, 2014–2021

For last 8 years, fatal nicotine intoxication cases have occurred constantly in Korea. Even though nicotine has been utilized for long time as a cigarette form, high concentration of liquid nicotine for e-cigarette became easily accessible after its global marketing in 2015. As a discrepancy of blood nicotine concentration between reported fatal cases and our postmortem cases was observed, we reviewed fatal nicotine poisoning cases in Seoul capital area from 2014 to 2021. All decedents in Seoul capital area from 2014 to 2021 whose cause of death was fatal nicotine poisoning were retrospectively studied; the route of administration included injection or ingestion of e-liquid, and attachment of nicotine transdermal patches. The autopsies were carried out at Seoul Institute of National Forensic Service (NFS), and heart bloods and/or femoral vein bloods were submitted for toxicological analysis. As a substitute of blood sample, liver tissues were collected and analyzed. The concentrations of nicotine from biological samples were quantitated with liquid chromatography-tandem mass spectrometry (LC-MS/MS) after sample preparation. The concentrations of nicotine were evaluated with several parameters; method of administration, co-administered drug or alcohol etc. Seventy-one fatal cases of nicotine were identified and analyzed. The average blood concentration of nicotine was 42 mg/L in heart blood (range: 0.69–250, n = 37) and 18 mg/L in femoral vein blood (range: 0.33–130, n = 64). The average nicotine concentration in liver tissue was 150 mg/kg (range: 1.6–610, n = 6). In a fatal case of poisoning with 70 nicotine transdermal patches, heart and femoral vein blood concentration of nicotine were 7.7 mg/L and 3.3 mg/L, respectively. The average femoral vein blood concentration was 19 mg/L in fatal nicotine cases with no other drug or alcohol co-administered (range: 1.27–87, n = 24), whereas that was 17 mg/L in fatal nicotine cases with drug(s) and/or alcohol co-administered (range: 0.33–130, n = 40). As a sub-group, the average femoral vein blood concentration was 11 mg/L in the cases with alcohol but no other drug co-administered (range: 0.33–58 mg/L, n = 16). In analysis of nicotine poisoning cases, the results should be interpreted with caution. Even though average value was presented here, the lethal blood concentration range was wide; from less than 1 mg/L to more than 100 mg/L. The tolerance at a certain concentration might be different whether the exposed person was a habitual smoker or a non-smoker. Also, co-administration of alcohol or drugs may increase the risk of death at lower nicotine concentration.

Determination of concentrations and toxicokinetics of triptolide in plasma and liver of mice by UHPLC-MS/MS

This study aims to establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS) method for determining the concentrations of triptolide(TP) in plasma and liver, and to explore the toxicokinetics of TP and the relationship between TP exposure and liver injury in C57 BL/6 mice, so as to provide reference for dissecting the toxicity mechanism of TP. The liquid chromatography was conducted with ZORBAX SB-C_(18) column(3.0 mm×100 mm, 3.5 μm) and the mobile phase of methanol-0.05 mmol·L~(-1) ammonium acetate. Electrospray ionization(ESI) and multiple reaction monitoring(MRM) mode were employed for mass spectrometry. After oral administration of TP(toxic dose 600 μg·kg~(-1)), the blood and liver tissues of the C57 BL/6 mice were collected at different time points to measure the TP concentrations in plasma and liver tissues. Furthermore, the blood biochemical indexes, including alkaline phosphatase(ALP), alanine aminotransferase(ALT), aspartate aminotransferase(AST), and total bile acid(TBA), were determined. After being processed by DAS 2.0, the experiment data showed that the TP in mice had the toxicokinetic parameters of T_(max)=5 min, C_(max)=14.38 ng·mL~(-1), t_(1/2)=0.76 h, AUC_(0-t)=5.63 h·ng·mL~(-1), MRT_(0-t)=0.56 h, and CL_(Z/F)=103.19 L·h~(-1)·kg~(-1). The trend of TP concentration in mouse liver tissue was consistent with that in plasma. The concentration of TP peaked at the time point of 5 min and then decreased until TP was completely metabolized. The plasma biochemical indexes(ALT, AST, ALP, and TBA) showed no significant changes within 3 h after TP administration. TP had high clearance rate and short residence time and did not significantly increase the blood biochemical indexes in mice. The results suggested that the exposure amount of free TP in vivo cannot directly cause liver injury, which might be caused by the binding of TP to some substances or the stimulation of inflammation and immune response.

Microplastics detected in cirrhotic liver tissue.

SummaryBackgroundThe contamination of ecosystem compartments by microplastics (MPs) is an ubiquitous problem. MPs have been observed in mice tissues, and recently in human blood, stool and placenta. However, two aspects remain unclear: whether MPs accumulate in peripheral organs, specifically in the liver, and if liver cirrhosis favours this process. We aimed to examine human liver tissue samples to determine whether MPs accumulate in the liver.MethodsThis proof-of-concept case series, conducted in Germany, Europe, analyzed tissue samples of 6 patients with liver cirrhosis and 5 individuals without underlying liver disease. A total of 17 samples (11 liver, 3 kidney and 3 spleen samples) were analyzed according to the final protocol. A reliable method for detection of MP particles from 4 to 30 µm in human tissue was developed. Chemical digestion of tissue samples, staining with Nile red, subsequent fluorescent microscopy and Raman spectroscopy were performed. Morphology, size and composition of MP polymers were assessed.FindingsConsidering the limit of detection, all liver, kidney and spleen samples from patients without underlying liver disease tested negative for MPs. In contrast, MP concentrations in cirrhotic liver tissues tested positive and showed significantly higher concentrations compared to liver samples of individuals without underlying liver disease. Six different microplastic polymers ranging from 4 to 30 µm in size were detected.InterpretationThis proof-of-concept case series assessed the presence of MPs in human liver tissue and found six different MP polymers in the liver of individuals with liver cirrhosis, but not in those without underlying liver disease. Future studies are needed to evaluate whether hepatic MP accumulation represents a potential cause in the pathogenesis of fibrosis, or a consequence of cirrhosis and portal hypertension.FundingNo funding was received for conducting this investigator driven study.

Dissecting the relationship between plasma and tissue metabolome in a cohort of women with obesity: Analysis of subcutaneous and visceral adipose, muscle, and liver.

Untargeted metabolomics of blood samples has become widely applied to study metabolic alterations underpinning disease and to identify biomarkers. However, understanding the relevance of a blood metabolite marker can be challenging if it is unknown whether it reflects the concentration in relevant tissues. To explore this field, metabolomic and lipidomic profiles of plasma, four sites of adipose tissues (ATs) from peripheral or central depot, two sites of muscle tissue, and liver tissue from a group of nondiabetic women with obesity who were scheduled to undergo bariatric surgery (n=21) or other upper GI surgery (n=5), were measured by liquid chromatography coupled with mass spectrometry. Relationships between plasma and tissue profiles were examined using Pearson correlation analysis subject to Benjamini-Hochberg correction. Plasma metabolites and lipids showed the highest number of significantly positive correlations with their corresponding concentrations in liver tissue, including lipid species of ceramide, mono- and di-hexosylceramide, sphingomyelin, phosphatidylcholine (PC), phosphatidylethanolamine (PE), lysophosphatidylethanolamine, dimethyl phosphatidylethanolamine, ether-linked PC, ether-linked PE, free fatty acid, cholesteryl ester, diacylglycerol and triacylglycerol, and polar metabolites linked to several metabolic functions and gut microbial metabolism. Plasma also showed significantly positive correlations with muscle for several phospholipid species and polar metabolites linked to metabolic functions and gut microbial metabolism, and with AT for several triacylglycerol species. In conclusion, plasma metabolomic and lipidomic profiles were reflective more of the liver profile than any of the muscle or AT sites examined in the present study. Our findings highlighted the importance of taking into consideration the metabolomic relationship of various tissues with plasma when postulating plasma metabolites marker to underlying mechanisms occurring in a specific tissue.

Anti-aging effect of phlorizin on D-galactose-induced aging in mice through antioxidant and anti-inflammatory activity, prevention of apoptosis, and regulation of the gut microbiota.

Aging is an inevitable and complicated process involving many physiological changes. Screening of natural biologically active anti-aging substances is a current research hotspot. Phlorizin (PZ), an important dihydrochalcone phytoconstituent, has been demonstrated to have antioxidant and anti-tumor effects. In this paper, different doses of PZ (20 and 40mg/kg) were used to research the protective effect on D-galactose (D-gal)-induced aging mice. Following hematoxylin and eosin staining and by observing the hippocampus, we found that PZ alleviated the damage caused by D-gal in neuronal cells, while PZ enhanced the learning and memory abilities of aging mice in a radical eight-arm maze. In order to explain the reasons for these anti-aging effects, we tested the antioxidant enzyme activity and malonic dialdehyde concentration in mouse serum, liver, and brain tissue. The contents of proteins related to anti-inflammation and apoptosis in brain tissue were analyzed, and the gut microbiota was also analyzed. The results indicated that PZ improved antioxidant enzyme activity while significantly reducing the malonic dialdehyde content. Western blotting analysis suggested that PZ effectively alleviated neuro-apoptosis via regulating the expressions of Bax, Bcl-2, and caspase-3. PZ also exerted anti-inflammation effects by regulating the interleukin-1β/inhibitor of nuclear factor kappa B alpha/nuclear factor kappa-light-chain-enhancer of activated B-cells signaling pathways in brain tissues. Importantly, PZ improved the structure and diversity of the gut microbiota, and the microbiota-gut-brain axis may hold a key role in PZ-induced anti-aging effects. In conclusion, PZ can be used as a potential drug candidate to combat aging.

Exercise Affects the Formation and Recovery of Alcoholic Liver Disease through the IL-6-p47phox Oxidative-Stress Axis.

(1) Background: To explore the effect of exercise on the formation and recovery of alcoholic liver disease (ALD) and whether the IL-6–p47phox oxidative–stress axis is involved in that process. (2) Methods: Firstly, 23 six-week-old male C57BL/6J mice were randomly divided into the Con group, ALD group, ALD + NOXI group, ALD + Ex group, and ALD + Ex + NOXI group. The Liber–DeCarli alcoholic liquid diet was used for 6 weeks to establish the ALD mice model, and the Con group was given the TP4030C control diet. The remaining groups were fed with the TP4030B alcoholic diet, and exercise intervention was started after the ALD model establishment and lasted for another 6 weeks, with or without administration of the NOX inhibitor apocynin by intraperitoneal injection on every exercise training day. Secondly, 28 mice were randomly divided into the Sed group, Eth group, Eth + Ex group and Eth + Ex + NOXI group. The Sed group was given the TP4030C control diet. The remaining groups were fed with the TP4030B alcoholic diet and exercise intervention was started synchronously combined with or without administration of intraperitoneal apocynin injections on every exercise training day for 5 weeks. After each individual experiment was accomplished, physiological assessment and biochemical analysis of blood and tissue samples were examined. (3) Results: The levels of TG in serum and IL-6 protein content in liver tissue in the ALD group were significantly increased compared to the Con group (p < 0.05); compared with ALD, p47phox expression in muscle was increased significantly in the ALD + NOXI group (p < 0.05), and TG in serum decreased in the ALD + Ex group (p < 0.05). TG in serum, AST/ALT ratio, and IL-6 content in both liver and muscle decreased (p < 0.05) in the ALD + Ex + NOXI group with MDA in muscle significantly increased (p < 0.01). The AST/ALT ratio, TG in serum, SOD in liver, and p47phox in both liver and muscle in the ALD + Ex + NOXI group were significantly decreased compared with the ALD + NOXI group (p < 0.01). Compared with the ALD + Ex group, the liver index and HDL-C levels in serum were decreased (p < 0.05) in the ALD + Ex + NOXI group. The degree of hepatocyte steatosis and inflammatory infiltration were ameliorated after exercise intervention. In the Eth group, the relative epididymal fat content, HDL-C level, and AST/ALT ratio were significantly decreased, and TG and gp91phox in liver were significantly higher than in the Sed group (p < 0.05, p < 0.01). Compared with the Eth group, the AST/ALT ratio, MDA in the liver, and NOX4 and p47phox protein expression in the liver were significantly increased, and body weight decreased significantly in the Eth + Ex group (p < 0.05, p < 0.01), as did TG in the liver and MDA in muscle. In the th + Ex + NOXI group, gp91phox expression in the liver and body weight were significantly decreased (p < 0.05, p < 0.01). In the Eth + Ex + NOXI group, the ratio of AST/ALT and MDA in muscle were increased when compared with the Eth + Ex group, and the protein expression of gp91phox and p47phox were much lower (p < 0.01). (4) Conclusions: 6 weeks of exercise intervention during the recovery phase of ALD ameliorates hepatocyte damage and dyslipidemia through the IL-6–p47phox oxidative–stress axis, and applying a NOX inhibitor in combination could optimize this. However, drinking alcohol during exercise exacerbates dyslipidemia and oxidative stress, with hepatocyte IL-6–p47phox downregulated.

Xanthophyll β-cryptoxanthin treatment inhibits hepatic steatosis without altering vitamin A status in β-carotene 9',10'-oxygenase knockout mice.

β-cryptoxanthin (BCX), one of the major carotenoids detected in human circulation, can protect against the development of fatty liver disease. BCX can be metabolized through β-carotene-15,15'-oxygenase (BCO1) and β-carotene-9',10'-oxygenase (BCO2) cleavage pathways to produce both vitamin A and apo-carotenoids, respectively, which are considered important signaling molecules in a variety of biological processes. Recently, we have demonstrated that BCX treatment reduced hepatic steatosis severity and hepatic total cholesterol levels in both wide type and BCO1-/-/BCO2-/- double knock out (KO) mice. Whether the protective effect of BCX is seen in single BCO2-/- KO mice is unclear. In the present study, male BCO2-/- KO mice at 1 and 5 months of age were assigned to two groups by age and weight-matching as follows: (I) -BCX control diet alone (AIN-93 purified diets); (II) +BCX 10 mg (supplemented with 10 mg of BCX/kg of diet) for 3 months. At 4 and 8 months of age, hepatic steatosis and inflammatory foci were evaluated by histopathology. Retinoids and BCX concentrations in liver tissue were analyzed by high-performance liquid chromatography (HPLC). Hepatic protein expressions of SIRT1, acetylated and total FoxO1, PGC1α, and PPARα were determined by the Western blot analysis. Real-time PCR for gene expressions (MCAD, SCD1, FAS, TNFα, and IL-1β gene expression relative to β-actin) was conducted in the liver. Steatosis was detected at 8 months but not at 4 months of age. Moreover, BCX supplementation significantly reduced the severity of steatosis in the livers of BCO2 KO mice, which was associated with changes in hepatic SIRT1 acetylation of FOXO1, PGC1α protein expression and PPARα protein expression in BCO2-/- KO mice. HPLC analysis showed that hepatic BCX was detected in BCX supplemented groups, but there were no differences in the hepatic levels of retinol and retinyl palmitate (RP) among all groups. The present study provided experimental evidence that BCX intervention can reduce liver steatosis independent of BCO2.

Population Pharmacokinetic-Pharmacodynamic Target Attainment Analysis of Flomoxef in the Serum and Liver Tissue of Patients Undergoing Hepatic Resection.

The purpose of this study was to investigate the population pharmacokinetics of prophylactic flomoxef based on serum and liver tissue concentrations and to demonstrate a pharmacodynamic target concentration in the serum and liver tissue exceeding the MIC in order to design an effective dosing regimen. Serum samples (n = 210) and liver tissue samples (n = 29) from 43 individuals were analyzed using a nonlinear mixed-effects model. The pharmacodynamics index target value was regarded as the probability of maintaining flomoxef serum trough and liver tissue concentrations exceeding the MIC90 values, 0.5 mg/L and 1.0 mg/L, for Escherichia coli and methicillin-susceptible Staphylococcus aureus, respectively. The final population pharmacokinetic model was a two-compartment model with linear elimination. Creatinine clearance (CLCR) was identified as a significant covariate influencing total clearance when CLCR was less than 60 mL/min. The probability of achieving concentrations in the serum and liver tissue exceeding the MIC90 for E. coli or methicillin-susceptible S. aureus for a 1 g bolus dose was above 90% at 2 h after the initial dose. Our findings suggest that population pharmacokinetic parameters are helpful for evaluating flomoxef pharmacokinetics and determining intraoperative flomoxef redosing intervals.

Qualitative and Quantitative Changes in Total Lipid Concentration and Lipid Fractions in Liver Tissue of Periparturient German Holstein Dairy Cows of Two Age Groups.

Fatty liver syndrome (FLS) is a common disease in high-producing dairy cows. Studies in humans suggest that the different hepatic lipid fractions play a role in this context. In dairy cows, little is known about the composition of fat stored in the liver, its periparturient dynamics, and the effect of cows' age. Therefore, our goal was to generate primary data in healthy cows to serve as reference values for future studies. Eight healthy German Holstein cows (2nd lactation, n = 3; ≥3rd lactation, n = 5) were examined 14 d antepartum and 7, 28, and 42 d postpartum. The examinations included clinical assessment, liver biopsy, blood sampling, and recording of milk yield. Total lipids (TL) in liver tissue were measured gravimetrically. The TL were separated into lipid fractions (triacylglycerol, TAG; phospholipids, PL; non-esterified fatty acids, NEFA; and cholesterol esters) using thin-layer chromatography, followed by gas chromatography for fatty acid determination. Concentrations of NEFA, ß-hydroxybutyrate, and cholesterol were analyzed in blood. Concentrations of TL, TAG, NEFA, and cholesterol esters in liver tissue and NEFA in blood increased in the periparturient period. The older cows had higher hepatic TL, TAG, and PL concentrations, higher relative hepatic concentrations of TAG in TL, higher NEFA concentrations in blood, a greater decrease in body condition, and higher milk yields between d 9 and 40 than the younger cows. We proposed that due to higher milk yield, older cows mobilized and deposited more fat in the liver, and the increase in hepatic TAG concentration was longer-lasting than in younger cows. Higher levels of structural lipids (PL) in older cows could be explained by higher demand for storage of TAG and cholesterol esters in lipid droplets or for the export of TAG via very-low-density lipoproteins. Results show that hepatic fat storage is a reversible process and does not necessarily cause clinical disease. Nevertheless, older cows have a more sustained and greater increase in hepatic TAG concentration, which may explain their increased risk of FLS. The results are limited in their extrapolation due to the small sample size and thereby possible selection bias but present a valuable basis for future studies.

Mechanism of dact2 gene inhibiting the occurrence and development of liver fibrosis

There are few reports about the relationship between dact2 and liver fibrosis. The inhibitory mechanism of dact2 in liver fibrosis is not clear, we need to further explore it. In this study has been shown that the dact2 gene can inhibit liver fibrosis. In this experiment, we used lentivirus as a vector to construct a lentivirus vector carrying the dact2 gene and packaged dact2 recombinant lentivirus and its control vector. HSC-T6 infected cells were observed. The effect of dact2 gene expression was activated by Wnt3a HSC-T6 cells. Immunoblot was used to detect α - SMA expression, TGF - β 1, Smad3, Smad7, β - Catenin and CyclinD1. The expression of MMP-2 and TIMP-1 was detected by real-time PCR. At the same time, dact2 recombinant lentivirus was injected into the tail vein. Carbon tetrachloride was used to establish the liver fibrosis model. After 7 weeks of modeling, the staining was used to observe the pathological changes of liver tissue, hydroxyproline was used to analyze the changes of collagen content in liver tissue, the expression of the protein was observed by immunohistochemistry, and the expression of fibrosis-related genes was detected by real-time PCR. Results showed that the dact2 gene expression could inhibit the activation of HSC-T6 cells and reduce the expression of TGF - β 1. The percentage of Smad3, β - Catenin and cyclinD1 protein was 50.02%, 46.73%, 47.58% and 37.50% respectively (P &lt; 0.05).

Protective Effect of Extract From America Ginseng and Hovenia dulcis Thunb Against Acute Alcohol-induced Liver Injury in Mice

Objective: The preventive effects of American ginseng and Hovenia dulcis Thunb (AGH) on alcohol-induced acute liver injury in mice. Methods: After one week of adaptation, ICR mice were randomly divided into normal group, model group and AGH (Ⅰ,Ⅱ,Ⅲ) groups. After giving the mice a continuous intragastric administration of AGH (100 mg/kg∙bw) for 14 days, all groups except the normal group were given three times of alcohol (5 g/kg) within 24 hours to induce liver damage. After the third alcohol administration for 6 hours, the mice were sacrificed. The activities of Aspartate aminotransferase (AST), Alanine aminotransferase (ALT) and the content of Triglycerides (TG) in serum were tested. The activity of Glutathione peroxidase (GSH-Px), Superoxide dismutase (SOD) and the contents of Glutathione (GSH) and Malondialdehyde (MDA) were detected. Pathological changes of liver were observed by H & E staining. The expression of p-ERK, p-JNK and p-P38 protein in liver tissue was detected by WB method. Results: Compared with model group, AGH could significantly reduce the activities of AST and ALT in serum (P<0.05), decreased the level of TG in serum (P<0.05) and MDA content in liver tissue (P<0.05). Meanwhile, AGH can also increase the activities of SOD and GSH-Px in liver tissue (P<0.05) and improve the level of GSH in liver tissue (P<0.05). Pathological observation showed that the main degeneration was fat drops, and some inflammatory cells were infiltrated in model group. AGH group had different degree of improvement on the pathological changes of the liver in mice. AHE significantly inhibited the expression of p-ERK, p-JNK and p-P38 (P<0.01) and inhibited the apoptosis of hepatocytes. Conclusion: AGH has obvious protective effect on acute alcohol-induced liver injury in mice.

Hepatoprotective Effects of Pomegranate Against Methotrexate-Induced Acute Liver Injury

Background: Methotrexate (MTX) is a synthetic antimetabolite with a wide range of clinical applications, but its liver toxicity induced mainly through oxidative stress represents a primary concern on its clinical use. Pomegranate fruit contains many polyphenolic compounds that possess potent antioxidant effects and, therefore, have a possible hepatoprotective effect. Objectives: This study seeks to address the hepatoprotective effects of pomegranate against MTX-induced liver injury. Materials and Methods: Twenty-eight healthy female albino mice were grouped into four groups; control and MTX groups received oral 0.5 ml distilled water, while; the PG150 group received 150mg/kg oral pomegranate, and the PG300 group received 300mg/kg oral pomegranate. The oral course continues for 10 days, and on the last day, all groups were injected with 20 mg/kg MTX intraperitoneally, except the control group injected with normal saline. 48-hours later, samples were collected and prepared for biochemical and histopathological evaluation. Results: After biochemical analysis, MTX causes an elevation in serum Alanine aminotransferase, Lactate dehydrogenase, and liver tissue malondialdehyde, indicating hepatic injury, while pomegranate pre-treatment will hold down this elevation significantly and dose-dependently, causing amelioration of the toxic effect of MTX; the histopathological findings support this finding. Also, MTX causes consumption of liver tissue content of superoxide dismutase (SOD) and Glutathione (GSH), while pomegranate pre-treatment boosts the SOD and GSH hepatic tissue level. Conclusion: Pomegranate has a dose-dependent amelioration effect on the toxic effect of MTX on the liver.

Effects of garlic extract on inflammatory cytokines in rats with type 1 and type 2 diabetes

Background and purposeOne of the most important complications of diabetes mellitus (DM) is liver damage. Hyperglycemia-induced inflammation is a major cause of diabetic complications. Therefore, compounds with hypoglycemic properties can be useful in controlling the complications of diabetes. The purpose of the current report was to evaluate the impact of garlic extract on inflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and transforming growth factor-β2 (TGF-β2) proteins levels in serum and liver tissue of diabetic rats. Experimental approachA total of thirty-six male Wistar rats were accidentally assigned to six classes: C: untreated group, D1: type 1 diabetes group (induced with streptozotocin, STZ), D1 + G: type 1 diabetes + garlic group (rats with diabetes type 1 that received garlic extract), D2: type 2 diabetes group (induced with STZ + niacinamide), D2 + G: type 2 diabetes + garlic group (rats with diabetes type 2 that received garlic extract), and G: garlic group (healthy rats that treated with garlic extract). After a period of five weeks, blood and tissue sections of animal's liver were obtained for serum and liver cytokines levels evaluation using specific ELISA kits. Findings/resultsThe serum and liver tissue TNF-α, IL-1β and TGF-β2 proteins concentrations were increased significantly in both groups with diabetes type 1 and type 2 compared with the untreated group (p < 0.01). The serum and liver tissue concentrations of the cytokines were attenuated in the diabetic rats treated with garlic extract (p < 0.05). Conclusion and implicationsThese findings indicated that garlic extract could act as potential medicinal purpose for diabetic complications through its beneficial effects in prevention of inflammation.

The Influence of Dietary Zinc Oxide Nanoparticles on Zootechnical Performance and Biometric Indices of Growing New Zealand White Rabbits

This study was designed to compare the effects of dietary conventional inorganic Zinc oxide (ZnO) and nano-zinc oxide (ZnO-NPs) on the zootechnical performance, selected blood serum indices, antioxidant status, general health and production of rabbit meat enriched with zinc enriched with nano-zinc in growing New Zealand White rabbits (NZW). A total of 481 newly-born rabbits produced from two natural successive mating between 36 primiparous (NZW) Rabbit does and 12 bucks by the intensive breeding system were utilized throughout this study. Rabbits does and bucks were randomly distributed into four equal groups, the control positive group G1 was fed the basal diet supplemented with premix contain 50 ppm inorganic zinc in the form of ZnO; while the control negative G2 was fed on a basal diet supplemented with Zinc Free premix, meanwhile, G3 was fed on the basal diet supplemented with premix contain 25 ppm nano zinc as zinc oxide nanoparticles, however, G4 was fed on the basal diet supplemented with 50 ppm nano zinc as zinc oxide nanoparticles in the premix, Litters after weaning continued on the same diet like their dams till the age of selling at 62 days old. The results showed that the growing rabbits in (G4) supplemented with 50 mg ZnO-NPs/kg diet surpassing significance (P≤ 0.05) in all groups and achieved the best result regarding (birth weight(g), weaning weight(g), sealing weight (g) moreover lower Pre-weaning mortality %, and Post-weaning mortality%). Also, G4 showed significant improvement in FCR in comparison with other groups. Furthermore, showed highest significant increase in total protein and globulin, HDL, SOD, GSH, Catalase activity, and zinc concentration in serum, liver, and thigh tissue and also showed the highest significant decrease in AG ratio, creatinine, urea, uric acid, cholesterol, Triglyceride, LDL, VLDL, risk factor, MDA and NO level in G4 at 62 day age old, compared to other groups, while there was no significant difference in albumin level between all the experimental groups. In conclusion, supplementation of 50 mg ZnO-Npskg diet in the premix to the diet of growing NZW rabbit maximize the productive performance and improve FCR without any adverse effect.

Anti- hyperglycemic activity of polyherbal formulation (Herbinsulplus) in experimental diabetes and its effects on key metabolic enzymes of carbohydrate metabolism

Diabetes mellitus is a metabolic disorder in which the body does not produce or properly utilize insulin, causes disturbances in carbohydrate, protein and lipid metabolism. In spite of the presence of series of known anti-diabetic medicine in the pharmaceutical market, remedies from medicinal plants are used with success to treat this disease. The investigation was undertaken to study the effect of polyherbal formulation on changes in Body weight, Plasma glucose, RBC, WBC, HB, Platelets, Hepatic glucokinase &amp; hexokinase, Glucose-6-phosphate and Glycogen content. In the current study, diabetes mellitus was induced by alloxan monohydrate at a dose of 150 mg/kg i.p to rats. A significant increase was observed in plasma glucose level in alloxan induced diabetic rats, whereas treatment with Glipizide (10mg/kg), polyherbal formulation (herbinsulplus) at two different doses (120mg/kg and 240mg/kg) showed significant antihyperglycemic activity in mild hyperglycemia. The administration of polyherbal formulation (herbinsulplus) at two doses showed an increase in body weight in alloxan-induced diabetic rats. Treatment with polyherbal formulation (herbinsul plus) at two doses prevented this alteration in glycogen content of liver tissue, but could not normalize the content of glycogen of the non-diabetic control. The groups treated with polyherbal formulation (herbinsul plus) (120mg/kg and 240mg/kg) and Glipizide (10mg/kg) showed greater persistence of islets of Langerhans and lesser degree of necrotic changes as compared to the untreated alloxan-induced diabetic rats. In conclusion, HerbinsulPlus exhibited significant anti-hyperglycemic activity in normal and alloxan induced diabetic rats and showed improvement in carbohydrate metabolizing enzymes as well as regeneration of β-cells of pancreas and so might be of value in diabetes treatment.

Plasma oxyphytosterols most likely originate from hepatic oxidation and subsequent spill-over in the circulation

We evaluated oxyphytosterol (OPS) concentrations in plasma and various tissues of two genetically modified mouse models with either increased cholesterol (apoE KO mice) or increased cholesterol and plant sterol (PS) concentrations (apoExABCG8 dKO mice). Sixteen female apoE KO and 16 dKO mice followed the same standard, low OPS-chow diet. Animals were euthanized at 36 weeks to measure PS and OPS concentrations in plasma, brain, liver and aortic tissue. Cholesterol and oxysterol (OS) concentrations were analyzed as reference for sterol oxidation in general. Plasma campesterol (24.1 ± 4.3 vs. 11.8 ± 3.0 mg/dL) and sitosterol (67.4 ± 12.7 vs. 4.9 ± 1.1 mg/dL) concentrations were severely elevated in the dKO compared to the apoE KO mice (p < 0.001). Also, in aortic and brain tissue, PS levels were significantly elevated in dKO. However, plasma, aortic and brain OPS concentrations were comparable or even lower in the dKO mice. In contrast, in liver tissue, both PS and OPS concentrations were severely elevated in the dKO compared to apoE KO mice (sum OPS: 7.4 ± 1.6 vs. 4.1 ± 0.8 ng/mg, p < 0.001). OS concentrations followed cholesterol concentrations in plasma and all tissues suggesting ubiquitous oxidation. Despite severely elevated PS concentrations, OPS concentrations were only elevated in liver tissue, suggesting that OPS are primarily formed in the liver and plasma concentrations originate from hepatic spill-over into the circulation.