Anti-CD163-dexamethasone conjugate inhibits the acute phase response to lipopolysaccharide in rats

AIM: To study the effect of a new anti-CD163-dexamethasone conjugate targeting activated macrophages on the hepatic acute phase response in rats. METHODS: Wistar rats were injected intravenous with either the CD163 targeted dexamethasone-conjugate(0.02 mg/kg) or free dexamethasone(0.02 or 1 mg/kg) 24 h prior to lipopolysaccharide(LPS)(2.5 mg/kg intraperitoneal). We measured plasma concentrations of tumour necrosis factor-a(TNF-a) and interleukin 6(IL-6) 2 h post-LPS and liver m RNAs and serum concentrations of the rat acute phase protein a-2-macroglobulin(a-2-M) 24 h after LPS. Also, plasma concentrations of alanine aminotransferase and bilirubin were measured at termination of the study. Spleen weight served as an indicator of systemic steroid effects.RESULTS: The conjugate halved the a-2-M liver m RNA(3.3 ± 0.6 vs 6.8 ± 1.1, P 0.01) and serum protein(201 ± 48 μg/mL vs 389 ± 67 μg/mL, P = 0.04) after LPS compared to low dose dexamethasone treated animals, while none of the free dexamethasone doses had an effect on liver m RNA or serum levels of a-2-M. Also, the conjugate reduced TNF-a(7208 ± 1977 pg/mL vs 21583 ± 7117 pg/mL, P = 0.03) and IL-6(15685 ± 3779 pg/mL vs 25715 ± 4036 pg/mL, P = 0.03) compared to the low dose dexamethasone. The high dose dexamethasone dose decreased the spleen weight(421 ± 11 mg vs 465 ± 12 mg, P 0.05) compared to controls, an effect not seen in any other group.CONCLUSION: Low-dose anti-CD163-dexamethasone conjugate effectively decreased the hepatic acute phase response to LPS. This indicates an anti-inflammatory potential of the conjugate in vivo.     

Obese diet-induced mouse models of nonalcoholic steatohepatitis-tracking disease by liver biopsy

AIM:To characterize development of diet-induced nonalcoholic steatohepatitis(NASH)by performing live biopsy in wild-type and genetically obese mice.METHODS:Male wild-type C57BL/6J(C57)mice(DIO NASH)and male Lep ob/Lep ob(ob/ob)mice(ob/ob-NASH were maintained on a diet high in trans-fat(40%)fructose(22%)and cholesterol(2%)for 26 and 12 wk respectively.A normal chow diet served as control in C57 mice(lean chow)and ob/ob mice(ob/ob chow)After the diet-induction period,mice were liver biopsied and a blinded histological assessment of steatosis and fibrosis was conducted.Mice were then stratified into groups counterbalanced for steatosis score and fibrosi stage and continued on diet and to receive daily PO dosing of vehicle for 8 wk.Global gene expression in liver tissue was assessed by RNA sequencing and bioin formatics.Metabolic parameters,plasma liver enzyme and lipids(total cholesterol,triglycerides)as well a hepatic lipids and collagen content were measured b biochemical analysis.Non-alcoholic fatty liver disease activity score(NAS)(steatosis/inflammation/ballooningdegeneration)and fibrosis were scored.Steatosis and fibrosis were also quantified using percent fractional area.RESULTS:Diet-induction for 26 and 12 wk in DIONASH and ob/ob-NASH mice,respectively,elicited progressive metabolic perturbations characterized by increased adiposity,total cholesterol and elevated plasma liver enzymes.The diet also induced clear histological features of NASH including hepatosteatosis and fibrosis.Overall,the metabolic NASH phenotype was more pronounced in ob/ob-NASH vs DIO-NASH mice.During the eight week repeated vehicle dosing period,the metabolic phenotype was sustained in DIO-NASH and ob/ob-NASH mice in conjunction with hepatomegaly and increased hepatic lipids and collagen accumulation.Histopathological scoring demonstrated significantly increased NAS of DIO-NASH mice(0 vs4.7±0.4,P0.001 compared to lean chow)and ob/ob-NASH mice(2.4±0.3 vs 6.3±0.2,P0.001compared to ob/ob chow),respectively.Furthermore,fibrosis stage was significantly elevated for DIO-NASH mice(0 vs 1.2±0.2,P0.05 compared to lean chow)and ob/ob NASH(0.1±0.1 vs 3.0±0.2,P0.001compared to ob/ob chow).Notably,fibrosis stage was significantly(P0.001)increased in ob/ob-NASH mice,when compared to DIO-NASH mice.CONCLUSION:These data introduce the obese dietinduced DIO-NASH and ob/ob-NASH mouse models with biopsy-confirmed individual disease staging as a preclinical platform for evaluation of novel NASH therapeutics.     

Significance of serum leptin and adiponectin levels in Egyptian patients with chronic hepatitis C virus associated hepatic steatosis and fibrosis

AIM: To study serum levels of leptin and adiponectin in patients with chronic hepatitis C virus infection genotype-4 (HCV-4) related steatosis and fibrosis. METHODS: We prospectively studied 45 untreated men with chronic HCV-4, with proven steatosis (group I, 30 patients), and fibrosis (group II, 15 patients), on liver biopsy. In addition, 15 healthy men (group III), matched for age, and body mass index were included. However, we excluded another five patients with steatohepatitis, and six patients with cirrhosis. We measured total serum leptin and adiponectin levels, as potential predictors for liver steatosis and fibrosis. Also, a correlation between these adipokines and various clinical and laboratory data were evaluated. All subjects were selected from Tropical and Internal medicine departments, Menoufiya University Hospital, Menoufiya, Egypt, during the period from February 2010 to August 2011. RESULTS: In group I, severity of hepatic steatosis was mild, moderate, and severe, in 19 patients (63.5%), 8 patients (26.5%), and 3 patients (10%), respectively. In contrast, in group II, hepatic fibrosis was found to be in stage 1, 2, and 3, in 6 patients (40%), in 6 patients (40%), and in 3 patients (20%), respectively. On comparing group I with group II, there was a significant decrease in serum adiponectin levels (131.4 ± 7.91 pg/mL vs 436 ± 9.75 pg/mL, P < 0.001), while there was no significant difference between both groups regarding serum leptin levels (34.69 ± 7.69 ng/mL vs 35.17 ± 1.06 ng/mL, P > 0.05). However, in the same group, when compared with group III, there was a significant increase in serum leptin levels (34.69 ± 7.69 ng/mL vs 10.69 ± 0.84 ng/mL, P < 0.001), while there was a significant decrease in serum adiponectin levels (131.4 ± 7.91 pg/mL vs 342.4 ± 44.48 pg/mL, P < 0.001). In contrast, in group II, when compared with group III, there was a significant increase in serum leptin and adiponectin levels (35.17 ± 1.06 ng/mL vs 10.69 ± 0.84 ng/mL, P < 0.001, and 436 ± 9.75 pg /mL vs 342.4 ± 44.48 pg/mL, P < 0.05, respectively), while there was no significant difference between both groups regarding serum creatinine (0.83 ± 0.34 vs 0.89 ± 0.24, P > 0.05). On the other hand, serum leptin was not correlated with serum adiponectin in group I and in group II (r = 0.09, P > 0.05, and r = -0.1, P > 0.05, respectively). However, serum adiponectin was significantly negatively correlated with serum aspartate transaminase in group I, but no correlation detected in group II (r =-0.39, P > 0.05, and r = -0.03, P > 0.05). CONCLUSION: In male patients with chronic HCV-4, serum adiponectin levels are elevated in hepatic fibrosis, but decreased in steatosis. Therefore, in contrast to leptin, adiponectin may be used as a non-invasive marker.     

Phosalone-induced inflammation and oxidative stress in the colon: evaluation and treatment

AIM: To investigate the side effects of phosalone on intestinal cells and to evaluate benefits of ellagic acid(EA) as a remedy.METHODS: In order to conduct an in vivo study, a rat model was used. The rats were divided into ten groups based on the materials used in the experiment and their dosage. The first group was fed normally. The second group was administered EA through gavage. Next Four groups were given(1/3, 1/5, 1/10, 1/20) LD50 phosalone; an organophosphorus compound. The last four groups received(1/3, 1/5, 1/10, 1/20) LD50 phosalone and of EA. After one month, the rats were sacrificed and their colon cells were examined to evaluate the level of inflammation, proteins and oxidative stress markers.RESULTS: The results of this research show that phosalone elevates oxidative stress and changes the level of tumor necrosis factor-a(TNF-α), interlukin-6β(IL-6β) and nuclear factor(NF)-κB proteins. EA administration reduced phosalone toxicity and changed oxidative stress and inflammatory markers for all phosalone doses. Overall changes in reduction of TNF-α(230.47 ± 16.55 pg/mg protein vs 546.43 ± 45.24 pg/mg protein, P 0.001), IL-6β(15.85 ± 1.03 pg/mg protein vs 21.55 ± 1.3 pg/mg protein, P 0.05), and NF-κB(32.47 ± 4.85 pg/mg protein vs 51.41 ± 0.71 pg/mg protein, P 0.05) manifest that the efficacy of EA is more viable for 1/3 LD50 dose of phosalone. Furthermore, EA is effective to counteract the negative outcomes of oxidative stress. When EA was used to treat 1/3 LD50 of phosalone's side effects, it improved the level of ACh E activity(48.5% ± 6% vs 25% ± 7%, P 0.05), TTM(0.391 ± 0.008 mmol/L vs 0.249 ± 0.032 mmol/L, P 0.05), FRAP(46.04 ± 5.005 μmol/L vs 18.22 ± 1.9 μmol/L, P 0.01) and MPO(0.222 ± 0.019 U/mg protein vs 0.387 ± 0.04 U/mg protein, P 0.05). CONCLUSION: This research highlights that EA is effective to alleviate the side effects of phosalone by reducing the level of oxidative stress and inflammatory proteins.     

Effect of Helicobacter pylori eradication on serum ghrelin and obestatin levels

AIM: To investigate changes in serum ghrelin and obestatin levels before and after Helicobacter pylori (H. pylori ) eradication. METHODS: A total of 92 patients presenting with symptoms of dyspepsia were enrolled in the study. Upper endoscopy was performed on all patients and used to diagnose H. pylori infection according to the presence of characteristic histopathological findings; seventy patients were diagnosed with H. pylori infection and the remaining 22 non-infected patients were classified as healthy controls. H. pylori eradication was accomplished by administering the classical triple therapy drug regimen, consisting of lansoprazole 30 mg bid , amoxicillin 1 g bid , and clarithromycin 500 mg tid for 14 d. The eradication of H. pylori was assessed with C14-urea breath test, which was performed at eight weeks after treatment. Levels of serum active ghrelin and obestatin were assessed at beginning of the study (prior to treatment) and after eight weeks. The levels were comparatively analyzed between the H. pylori negative control group, the H. pylori eradicated group, and the H. pylori non-eradicated group. RESULTS: A total of 92 patients, 50 females and 42 males with a mean age of 38.2 ± 11.9 years (range: 19-64), were analyzed. H. pylori eradication success was achieved in 74.3% (52/70) of H. pylori positive patients. The initial levels of ghrelin in the H. pylori positive and control cases were 63.6 ± 19.8 pg/mL and 65.1 ± 19.2 pg/mL (P=0.78), respectively, and initial obestatin levels were 771±427 pg/mL and 830 ± 296 pg/mL (P=0.19), respectively. The difference between the initial levels and the week 8 levels of ghrelin and obestatin in the control group was insignificant [4.5% (P=0.30) and -0.9% (P=0.65), respectively]. The difference between the initial and week 8 levels of ghrelin and obestatin in the H. pylori non-eradicated group were also insignificant [0.9% (P=0.64) and 5.3% (P=0.32), respectively]. The H. pylori eradicated group had a greater change in obestatin levels when compared to the     

Endothelial nitric oxide synthase content in adipose tissue from obese and lean African American and white American women

It has been demonstrated that the enzyme endothelial nitric oxide synthase (eNOS) is present in adipose tissue, resulting in nitric oxide production and subsequent inhibition of lipolysis. A higher eNOS content has also been reported in the subcutaneous abdominal adipose tissue of obese than in that of lean white men. Furthermore, a lower lipolytic rate in obese than in lean women and a lower lipolytic rate in African American (AA) than in white American (WA) women have been demonstrated. The purpose of this study was to determine if eNOS protein content is higher in the subcutaneous and omental adipose tissues of obese than in those of lean women and if eNOS protein content is higher in the subcutaneous and omental adipose tissues of AA than in those of WA women. Whole tissue homogenates were prepared from frozen omental and subcutaneous adipose tissue samples obtained from lean and obese and AA and WA elective abdominal surgery patients and were analyzed for eNOS protein content using enzyme-linked immunosorbent assay. The adipose tissue eNOS protein content was approximately 40% higher in obese than in lean individuals (omental, 326.9 +/- 40.5 pg/mL lean and 445.3 +/- 38.0 pg/mL obese; subcutaneous, 246.8 +/- 20.8 pg/mL lean and 343.1 +/- 19.0 pg/mL obese; P < .05). There was no difference between the races for eNOS protein content in omental adipose tissue. In subcutaneous adipose tissue, there was a higher eNOS content in obese (417.1 +/- 78.9 pg/mg total protein) than in lean (216.7 +/- 29.9 pg/mg total protein) (P < .05) WA women, but there was no difference in subcutaneous adipose eNOS content between obese and lean AA women (250.7 +/- 47.4 and 294.1 +/- 42.2 pg/mg total protein, respectively). The higher eNOS content in the adipose tissue of obese than in that of lean WA women in the fasted state may contribute to the reduced lipolytic activity in WA women; however, eNOS protein content probably does not contribute to differences in lipolytic rates between AA and WA women.     

Gardenia jasminoides attenuates hepatocellular injury and fibrosis in bile duct-ligated rats and human hepatic stellate cells

AIM:To investigate the anti-hepatofibrotic effects of Gardenia jasminoides in liver fibrosis.METHODS:Male Sprague-Dawley rats underwent common bile duct ligation(BDL) for 14 d and were treated with Gardenia jasminoides by gavage.The ef-fects of Gardenia jasminoides on liver fibrosis and the detailed molecular mechanisms were also assessed in human hepatic stellate cells(LX-2) in vitro.RESULTS:Treatment with Gardenia jasminoides decreased serum alanine aminotransferase(BDL vs BDL + 100 mg/kg Gardenia jasminoides,146.6 ± 15 U/L vs 77 ± 6.5 U/L,P = 0.0007) and aspartate aminotransferase(BDL vs BDL + 100 mg/kg Gardenia jasminoides,188 ± 35.2 U/L vs 128 ± 19 U/L,P = 0.005) as well as hydroxyproline(BDL vs BDL + 100 mg/kg Gardenia jasminoides,438 ± 40.2 μg/g vs 228 ± 10.3 μg/g liver tissue,P = 0.004) after BDL.Furthermore,Gardenia jasminoides significantly reduced liver mRNA and/or protein expression of transforming growth factor β1(TGF-β1),collagen type?Ⅰ?(Col?Ⅰ) and α-smooth muscle actin(α-SMA).Gardenia jasminoides significantly suppressed the upregulation of TGF-β1,Col?Ⅰand α-SMA in LX-2 exposed to recombinant TGF-β1.Moreover,Gardenia jasminoides inhibited TGF-β1-induced Smad2 phosphorylation in LX-2 cells.CONCLUSION:Gardenia jasminoides exerts antifibrotic effects in the liver fibrosis and may represent a novel antifibrotic agent.     

Antifibrotic effects of ambrisentan,an endothelin-A receptor antagonist,in a non-alcoholic steatohepatitis mouse model

AIM: To examine the effects of the endothelin type A receptor antagonist ambrisentan on hepatic steatosis and fibrosis in a steatohepatitis mouse model.METHODS: Fatty liver shionogi(FLS) FLS-ob/ob mice(male, 12 wk old) received ambrisentan(2.5 mg/kg orally per day; n = 8) or water as a control(n = 5) for 4 wk. Factors were compared between the two groups, including steatosis, fibrosis, inflammation, and endothelin-related gene expression in the liver.RESULTS: In the ambrisentan group, hepatic hydroxyproline content was significantly lower than in the control group(18.0 μg/g ± 6.1 μg/g vs 33.9 μg/g ± 13.5 μg/g liver, respectively, P = 0.014). Hepatic fibrosis estimated by Sirius red staining and areas positive for α-smooth muscle actin, indicative of activated hepatic stellate cells, were also significantly lower in the ambrisentan group(0.46% ± 0.18% vs 1.11% ± 0.28%, respectively, P = 0.0003; and 0.12% ± 0.08% vs 0.25% ± 0.11%, respectively, P = 0.047). Moreover, hepatic RNA expression levels of procollagen-1 and tissue inhibitor of metalloproteinase-1(TIMP-1) were significantly lower by 60% and 45%, respectively, in the ambrisentan group. Inflammation, steatosis, and endothelin-related m RNA expression in the liver were not significantly different between the groups.CONCLUSION: Ambrisentan attenuated the progression of hepatic fibrosis by inhibiting hepatic stellate cell activation and reducing procollagen-1 and TIMP-1 gene expression. Ambrisentan did not affect inflammation or steatosis.     

Pharmacology of novel Chinese medicines screened for treatment of severe acute pancreatitis

AIM: To screen the enzyme-inhibition and observe the pharmacologic effects of circulation-improving Chinese medicines on intestinal and pancreatic hemodynamics. METHODS: In vitro screening of the amylase-and lipase-inhibiting effects of nine Chinese traditional medicines was carried out. Each extract was prepared following methods described by LIN Qi-Shou. Tests of the inhibition of trypsin and elastase activity were carried out by following methods described by ZHANG Tian-Min s and JIANG Chuan-Kui. A Modified Leslie method was used determine the effects of the best circulation-improving Chinese herb, Tao-Ren (Semen Persicae), and its extract (HHI-I), on intestinal hemodynamics. An electromagnetic flowmeter was used to measure intestinal blood flow and and a blood oxygen meter was used to measure oxygen consumption. A laser Doppler microcirculation dynamic analyzer was used for to measurement the effect of HHI-I on pancreatic microcirculation, and a tissue oxygen meter was used to measure changes of pancreatic oxygen partial pressure. RESULTS: Of the nine Chinese traditional medicines, Yuan-Hu (Rhizome Corydalis) had the most potent inhibitory activity on both amylase and lipase activity, followed by Da Huang, Zhi Zi, Huang Qin, and Hang Shao. Canine experiments found that Huoxue Huayu (HHI-I) improved intestinal hemodynamics. Intestinal blood flow 20 min after infusion of HHI-I, was 225.0 ± 68.51 mL/min, which was significantly higher than that before infusion (201.34 ± 70.21 mL/min, P < 0.05). Blood flow increased to 245.40 ± 82.78 mL/min (P < 0.05) at 40 min and 252.20 ± 82.41 mL/min (P < 0.01). 60 min after infusion. Intestinal oxygen consumption also increased significantly, to 5.33 ± 2.57 mL/min at 60 min after infusion, compared with (2.72 ± 1.09 mL/min (P < 0.05) at baseline. HHI-I improved pancreatic microcirculation and tissue oxygen partial pressure. The basal microcirculatory blood flow of 20.4 ± 5.0 mL/min/100 g body weight increased to 49.0 ± 9.0 mL/min/100g body weight 20 min after administration (P < 0.05). Blood flow increase further to 51.0 ± 7.97 mL/min/100 g body weight (P < 0.01) at 40 min and 54.8 ± 15.51 mL/min/100 g body weight (P < 0.01) at 60 min. Pancreatic oxygen partial pressure was 6.21 ± 0.94 kPa at baseline, rose to 7.55 ± 1.40 kPa at 20 min (P < 0.01), 7.65 ± 1.76 kPa (P < 0.05) at 40 min, and 7.67 ± 1.64 kPa (P < 0.01) at 60 min after HHI-I administration. In the control group, saline administration had no significant effect of the indices. CONCLUSION: Yuan-Hu and Tao-Ren used together have the potential to become novel, effective medicines     

Comparative effects of &alpha;2&delta;-1 ligands in mouse models of colonic hypersensitivity

AIM: To investigate anti-hypersensitive effects of α₂δ-1 ligands in non-inflammatory and inflammation-associated colonic hypersensitivity (CHS) mouse models. METHODS: To induce an inflammation-associated CHS, 1% dextran sulfate sodium (DSS) was administered to C57Bl/6J male mice, in drinking water, for 14 d. Regarding the non-inflammatory neonatal maternal separation (NMS) -induced CHS model, wild-type C57BI/6J pups were isolated from their mother from day 2 to day 14 (P2 to P14), three hours per day (from 9:00 a.m. to 12:00 p.m.). Colorectal distension was performed by inflating distension probe from 20 μL to 100 μL by 20 μL increment step every 10 s. After a first colorectal distension (CRD), drugs were administered subcutaneously, in a cumulative manner, (Gabapentin at 30 mg/kg and 100 mg/kg; Pregabalin at 10 mg/kg and 30 mg/kg; Carbamazepine at 10 mg/kg and 30 mg/kg) and a second CRD was performed one hour after each injection. RESULTS: The visceromotor response (VMR) to CRD was increased by our NMS paradigm protocol in comparison to non-handled (NH) mice, considering the highest distension volumes (80 μL: 0.783 ± 0.056 mV/s vs 0.531 ± 0.034 mV/s, P < 0.05 and 100 μL: 1.087 ± 0.056 mV/s vs 0.634 ± 0.038 mV/s, P < 0.05 for NMS and NH mice, respectively). In the inflammation-associated CHS, DSS-treated mice showed a dramatic and significant increase in VMR at 60 and 80 μL distension volumes when compared to control mice (60 μL: 0.920 ± 0.079 mV/s vs 0.426 ± 0.100 mV/s P < 0.05 and 80 μL: 1.193 ± 0.097 mV/s vs 0.681 ± 0.094 mV/s P < 0.05 for DSS- and Water-treated mice, respectively). Carbamazepine failed to significantly reduce CHS in both models. Gabapentin significantly reduced CHS in the DSS-induced model for both subcutaneous injections at 30 or 100 mg/kg. Pregabalin significantly reduced VMR to CRD in the non-inflammatory NMS-induced CHS model for the acute subcutaneous administration of the highest cumulative dose (30 mg/kg) and significantly reduced CHS in low-dose DSS-treated mice in a dose-dependent manner. Finally, the percent decrease of AUC induced by acute GBP or Pregabalin treatment were higher in the inflammatory DSS-induced CHS model in comparison to the non-inflammatory NMS-induced CHS model. CONCLUSION: This preclinical study demonstrates α₂δ-1 ligands efficacy on inflammation-associated CHS, highlighting their potential clinical interest in patients with chronic abdominal pain and moderate intestinal inflammation.     

小檗碱通过调节肠道菌群改善高脂饮食诱导的非酒精性脂肪性肝病大鼠肝组织炎症实验研究

目的 探讨应用小檗碱(BBR)处理对高脂饮食诱导的非酒精性脂肪性肝病(NAFLD)大鼠肝组织炎症和肠道菌群的影响。方法 将45只SD大鼠随机分为对照组、模型组和小檗碱处理组,每组15只。在模型组和小檗碱处理组,采用高脂饮食喂养诱导NAFLD模型。实验结束时取肝组织行病理学检查,取血清检测空腹血糖(FPG)和空腹胰岛素(FINS),计算胰岛素抵抗指数(IRI);采集粪便,采用16SrRNA序列分析法检测肠道菌群变化;采用ELISA法检测血清白细胞介素-6(IL-6)、IL-10和肿瘤坏死因子-α(TNF-α)水平。结果 在高脂喂养16 w末,取2只动物肝组织检查,提示造模成功。在实验结束时,小檗碱处理组动物肝组织学病变明显改善;小檗碱处理组大鼠血清ALT、AST和LDL水平分别为(78.0±6.0)IU/L、(119.6±8.8)IU/L和(61.3±5.0)mg/dL,显著低于模型组【分别为(211.5±9.0)IU/L、(312.6±9.7)IU/L和(97.6±8.9)mg/dL,P<0.05】,而血清HDL水平为(70.0±5.4)mg/dL,显著高于模型组【(14.6±5...     

乙型肝炎肝硬化患者外周血dNLR、MLR和SII变化及其临床意义分析

目的 探讨乙型肝炎肝硬化患者外周血衍生中性粒细胞/淋巴细胞比值(dNLR)、单核细胞/淋巴细胞比值(MLR)和系统性免疫性炎症指数(SII)变化及其临床意义。方法 2019年1月～2021年12月我院收治的乙型肝炎肝硬化患者85例和慢性乙型肝炎(CHB)患者85例,采用ELISA法检测血清肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)和IL-17。常规检测血细胞计数。结果 乙型肝炎肝硬化患者血清TBIL和INR分别为(43.1±8.5)μmol/L和(1.3±0.6),显著高于【分别为(19.4±3.0)μmol/L和(1.1±0.2),P<0.05】,而血清ALT和ALB水平分别为(63.6±8.2)U/L和(30.8±4.6)g/L,显著低于CHB组【分别为(104.1±14.9)U/L和(39.0±8.1)g/L,P<0.05】;肝硬化患者血清TNF-α、IL-6和IL-17水平分别为(88.7±11.6)pg/mL、(95.6±12.5)pg/mL和(45.6±8.9)ng/mL,显著高于CHB组【分别为(68.2±9.3)pg/mL、(67.9±9.5)p...     

Increased duodenal expression of miR-146a and -155 in pediatric Crohn&rsquo;s disease

AIM: To evaluate the role of microRNA (miR)-146a, -155 and -122 in the duodenal mucosa of pediatric patients with Crohn’s disease (CD) and the effect of transforming growth factor-β (TGF-β) on these miRs in duodenal epithelial and fibroblast cells. METHODS: Formalin-fixed, paraffin-embedded biopsies derived from the macroscopically inflamed (CD inflamed: n = 10) and intact (CD intact: n = 10) duodenal mucosa of pediatric CD patients and control children (C: n = 10) were examined. Expression of miR-146a, -155 and -122 was determined by real-time polymerase-chain reaction (PCR). The expression of the above miRs was investigated in recombinant human TGF-β (1 nmol/L, 24 h) or vehicle treated small intestinal epithelial cells (CCL-241) and primary duodenal fibroblast cells derived from healthy children as well. RESULTS: Expression of miR-146a was significantly higher in the inflamed duodenal mucosa compared to the intact duodenal mucosa of children with CD (CD inflamed: 3.21 ± 0.50 vs CD intact: 0.62 ± 0.26, P≤ 0.01) and to the control group (CD inflamed: 3.21 ± 0.50 vs C: 1.00 ± 0.33, P≤ 0.05). The expression of miR-155 was significantly increased in the inflamed region of the duodenum compared to the control group (CD inflamed: 4.87 ± 1.02 vs Control: 1.00 ± 0.40, P≤ 0.001). The expression of miR-122 was unchanged in the inflamed or intact mucosa of CD patients compared to controls. TGF-β treatment significantly decreased the expression of miR-155 in small intestinal epithelial cells (TGF-β: 0.7 ± 0.083 vs Control: 1 ± 0.09, P≤ 0.05) and also the expression of miR-146a (TGF-β: 0.67 ± 0.04 vs Control: 1 ± 0.15, P≤ 0.01) and miR-155 (TGF-β: 0.72 ± 0.09 vs Control: 1 ± 0.06, P≤ 0.05) in primary duodenal fibroblasts compared to corresponding vehicle treated controls. TGF-β treatment did not influence the expression of miR-122. CONCLUSION: The elevated expression of miR-146a and -155 in the inflamed duodenal mucosa of CD patients suggests the role of these miRs in the pathomechanism of inflammatory bowel disease. Anti-inflammatory TGF-β plays an important role in the regulation of the expression of these miRs.     

Serum adipokines might predict liver histology findings in non-alcoholic fatty liver disease

AIM: To assess significance of serum adipokines to determine the histological severity of non-alcoholic fatty liver disease.METHODS: Patients with persistent elevation in serum aminotransferase levels and well-defined characteristics of fatty liver at ultrasound were enrolled. Individuals with a history of alcohol consumption, hepatotoxic medication, viral hepatitis or known liver disease were excluded. Liver biopsy was performed to confirm nonalcoholic liver disease(NAFLD). The degrees of liver steatosis, lobular inflammation and fibrosis were determined based on the non-alcoholic fatty liver activity score(NAS) by a single expert pathologist. Patients with a NAS of five or higher were considered to have steatohepatitis. Those with a NAS of two or lower were defined as simple fatty liver. Binary logistic regression was used to determine the independent association of adipokines with histological findings. Receiver operating characteristic(ROC) analysis was employed to determine cut-off values of serum adipokines to discriminate the grades of liver steatosis,lobular inflammation and fibrosis. RESULTS: Fifty-four participants aged 37.02 ± 9.82 were enrolled in the study. Higher serum levels of visfatin, IL-8, TNF-α levels were associated independently with steatosis grade of more than 33% [β = 1.08(95%CI: 1.03-1.14), 1.04(95%CI: 1.008-1.07), 1.04(95%CI: 1.004-1.08), P 0.05]. Elevated serum IL-6 and IL-8 levels were associated independently with advanced lobular inflammation [β = 1.4(95%CI: 1.09-1.8), 1.07(95%CI: 1.003-1.15), P 0.05]. Similarly, higher TNF-α, resistin, and hepcidin levels were associated independently with advanced fibrosis stage [β = 1.06(95%CI: 1.002-1.12), 19.86(95%CI: 2.79-141.19), 560.72(95%CI: 5.98-5255.33), P 0.05]. Serum IL-8 and TNF-α values were associated independently with the NAS score, considering a NAS score of 5 as the reference value [β = 1.05(95%CI: 1.01-1.1), 1.13(95%CI: 1.04-1.22), P 0.05]. CONCLUSION: Certain adipokines may determine the severity of NAFLD histology accurately.     

Steatotic livers are susceptible to normothermic ischemia-reperfusion injury from mitochondrial Complex-I dysfunction

AIM: To assess the effects of ischemic preconditioning (IPC, 10-min ischemia/10-min reperfusion) on steatotic liver mitochondrial function after normothermic ischemia-reperfusion injury (IRI). METHODS: Sixty male Sprague-Dawley rats were fed 8-wk with either control chow or high-fat/high-sucrose diet inducing > 60% mixed steatosis. Three groups (n = 10/group) for each dietary state were tested: (1) the IRI group underwent 60 min partial hepatic ischemia and 4 h reperfusion; (2) the IPC group underwent IPC prior to same standard IRI; and (3) sham underwent the same surgery without IRI or IPC. Hepatic mitochondrial function was analyzed by oxygraphs. Mitochondrial Complex-I, Complex-II enzyme activity, serum alanine aminotransferase (ALT), and histological injury were measured. RESULTS: Steatotic-IRI livers had a greater increase in ALT (2476 ± 166 vs 1457 ± 103 IU/L, P < 0.01) and histological injury following IRI compared to the lean liver group. Steatotic-IRI demonstrated lower Complex-I activity at baseline [78.4 ± 2.5 vs 116.4 ± 6.0 nmol/(min.mg protein), P < 0.001] and following IRI [28.0 ± 6.2 vs 104.3 ± 12.6 nmol/(min.mg protein), P < 0.001]. Steatotic-IRI also demonstrated impaired Complex-I function post-IRI compared to the lean liver IRI group. Complex-II activity was unaffected by hepatic steatosis or IRI. Lean liver mitochondrial function was unchanged following IRI. IPC normalized ALT and histological injury in steatotic livers but had no effect on overall steatotic liver mitochondrial function or individual mitochondrial complex enzyme activities. CONCLUSION: Warm IRI impairs steatotic liver Complex-I activity and function. The protective effects of IPC in steatotic livers may not be mediated through mitochondria.     

Desferrioxamine in warm reperfusion media decreases liver injury aggravated by cold storage

AIM:To evaluate whether desferrioxamine decreases ischemia and perfusion injury aggravated by cold storage(CS)in a rat liver perfusion model. METHODS:Isolated rat livers were kept in CS in University of Wisconsin Solution for 20 h at 4℃,then exposed to 25 min of warm ischemia(WI)at 37℃ followed by 2 h of warm perfusion(WP)at 37℃with oxygenated(95%oxygen and 5%carbon dioxide) Krebs-Henseleit buffer.Desferrioxamine(DFO),an iron chelator,was added at different stages of storage,ischemia and perfusion:in CS only,in WI only,in WP only, in WI and perfusion,or in all stages.Effluent samples were collected after CS and after WI.Perfusate samples and bile were collected every 30 min(0,0.5,1,1.5 and 2 h)during liver perfusion.Cellular injury was assessed by the determination of lactate dehydrogenase(LDH) and aspartate aminotransferase(AST)in the effluent and perfusate samples.Total iron was analysed in the perfusate samples.After WP,the liver was collected for the determination of liver swelling(wet to dry ratio) and liver morphological examination(hematoxylin and eosin staining). RESULTS:Increased CS time caused increased liver dysfunction during WP.After 2 h of WP,liver injury was indicated by increased release of AST(0.5 h CS:9.4± 2.2 U/g liver vs 20 h CS:45.9±10.8 U/g liver,P<0.05) and LDH(0.5 h CS:59±14 U/g liver vs 20 h CS:297 ±71 U/g liver,P<0.05).There was an associated increase in iron release into the perfusate(0.5 h CS:0.11 ±0.03μmoL/g liver vs 20 h CS:0.58±0.10μmoL/g liver,P<0.05)and reduction in bile flow(0.5 h CS: 194±12μL/g vs 20 h CS:71±8μL/g liver,P<0.05). When DFO was added during WI and WP following 20 h of CS,release of iron into the perfusate was de- creased(DFO absent 0.58±0.10μmoL/g liver vs DFO present 0.31±0.06μmoL/g liver,P<0.05),and liver function substantially improved with decreased release of AST(DFO absent 45.9±10.8 U/g liver vs DFO present 8.1±0.9 U/g liver,P<0.05)and LDH(DFO absent 297±71 U/g liver vs DFO present 56±7 U/g liver,P<0.05),and inc     

三维适形放射治疗不同大体肿瘤体积的原发性肝癌患者疗效研究*

目的 探讨不同大体肿瘤体积(GTV)的原发性肝癌(PLC)患者放射治疗后生存情况。方法 2018年1月～2020年12月我院收治的PLC患者75例,根据GTV的不同将患者分为A组(＜125 cm³)20例、B组(126～999 cm³)37例和C组(＞1000 cm³)18例,均接受三维适形放射治疗(3DCRT)。采用ELISA法检测血清甲胎蛋白(AFP)、甲胎蛋白异质体L3(AFP-L3)、高尔基体糖蛋白73(GP73)、血管内皮生长因子(VEGF)和基质金属蛋白酶(MMP)水平。结果 在治疗后4 w末,A组客观缓解率(ORR)为70.0%,显著高于B组的51.4%或C组的33.3%(P＜0.05);A组血清AFP、AFP-L3、GP73、VEGF和MMP水平分别为(121.0±20.8)ng/mL、(5.9±0.9)ng/mL、(62.8±3.5)ng/mL、(265.3±20.4)pg/mL和(60.2±11.7)ng/L,显著低于B组【分别为(133.0±18.1)ng/mL、(6.5±0.7)ng/mL、(71.4±3.2)ng/mL、(296.1±30.9)pg/mL和(72.5±7.3)ng/L,P＜0.05】或C组【分别为(143.0±15.3)ng/mL、(6.9±0.8)ng/mL、(78.3±3.7)ng/mL、(320.5±32.7)pg/mL和(85.5±15.6)ng/L,P＜0.05】;A组1 a累积生存率为72.2%(13/18),显著高于B组的52.9%(18/34)或C组的41.2%(7/17,P＜0.05)。结论 采用3DCRT治疗PLC患者近期有效,但需考虑GTV的影响,GTV越小,近期疗效越好,值得进一步研究。     

抑制TRAF6基因表达对自身免疫性肝炎大鼠肝组织NF-κB信号通路相关蛋白表达的影响

目的 探讨抑制TRAF6基因表达对自身免疫性肝炎(AIH)大鼠肝组织NF-κB信号通路相关蛋白表达的影响。方法 随机将40只SD大鼠分为对照组、模型组、空载体组和TRAF6 shRNA干扰组,每组10只,采用特异性抗原S-100诱导建立大鼠AIH模型,分别给予生理盐水、转染空载体和转染TRAF6 shRNA处理。采用免疫组化法检测肝组织TRAF6蛋白表达,采用qRT-PCR法检测肝组织TRAF6 mRNA水平,采用ELISA法检测肝组织匀浆白介素1β(IL-1β)和白介素6(IL-6)水平,采用Western blot检测肝组织NF-κB信号通路相关基因蛋白表达。结果 模型组大鼠肝组织TRAF6 mRNA水平为(6.2±0.4),显著高于正常组(1.0±0.2,P＜0.05),而TRAF6 shRNA干预组大鼠肝组织TRAF6 mRNA水平为(1.5±0.2),较模型组显著降低(P＜0.05);模型组大鼠肝组织匀浆IL-1β水平为(60.3±8.1)pg/mL,IL-6水平为(41.5±5.9)pg/mL,均显著高于正常组[(8.9±0.6)pg/mL和(15.6±0.6)pg/mL, P＜0.05];模型组大鼠肝组织IκBα、NF-κB p65、NF-κB p50、p-NF-κB p65和p-NF-κB p50蛋白表达相对水平分别为(1.3±0.1)、(1.6±0.1)、(1.3±0.1)、(1.3±0.1)和(1.1±0.1),均显著高于正常组[分别为(0.3±0.03)、(0.3±0.02)、(0.3±0.03)、(0.3±0.03)和(0.3±0.03),P＜0.05],而抑制了TRAF6表达组大鼠肝组织IL-1β为(31.5±4.0)pg/mL,IL-6为(27.4±4.2)pg/mL,IκBα、NF-κB p65、NF-κB p50、p-NF-κB p65和p-NF-κB p50蛋白表达分别为(1.0±0.1)、(0.9±0.05)、(0.7±0.1)、(0.8±0.1)和(0.5±0.05),均显著低于模型组(均P＜0.05)。结论 抑制TRAF6基因表达可降低AIH大鼠肝内炎症水平,从而能改善肝组织损伤,其机制可能与调节了NF-κB信号通路的活化有关。