Morphological aspects of small-duct cholangiopathies:A minireview

The biliary system consists of intrahepatic and extrahepatic bile ducts lined by biliary epithelial cells(cholangiocytes).Bile ducts and cholangiocytes are affected by a variety of disorders called cholangiopathies,which differ in aetiology,pathogenesis,and morphology.Classification of cholangiopathies is complex and reflects pathogenic mechanisms(immune-mediated,genetic,drug-and toxininduced,ischaemic,infectious,neoplastic),predominant morphological patterns of biliary injury(suppurative and non-suppurative cholangitis,cholangiopathy),and specific segments of the biliary tree affected by the disease process.While the involvement of large extrahepatic and intrahepatic bile ducts is typically visualised using radiology imaging,histopathological examination of liver tissue obtained by percutaneous liver biopsy still plays an important role in the diagnosis of cholangiopathies affecting the small intrahepatic bile ducts.To increase the diagnostic yield of a liver biopsy and determine the optimal therapeutic approach,the referring clinician is tasked with interpreting the results of histopathological examination.This requires knowledge and understanding of basic morphological patterns of hepatobiliary injury and an ability to correlate microscopic findings with results obtained by imaging and laboratory methods.This minireview describes the morphological aspects of small-duct cholangiopathies pertaining to the diagnostic process.

Regulated upon activation,normal T cell expressed and secreted(RANTES)levels in the peripheral blood of patients with Alzheimer’s disease

Alzheimer’s disease(AD)is the most common type of dementia,but it is very difficult to diagnose with certainty,so many AD studies have attempted to find early and relevant diagnostic markers.Regulated upon activation,normal T cell expressed and secreted(RANTES,also known as C-C chemokine ligand)is a chemokine involved in the migration of T cells and other lymphoid cells.Changes in RANTES levels and its expression in blood or in cerebrospinal fluid have been reported in some neurodegenerative diseases,such as Parkinson’s disease and multiple sclerosis,but also in metabolic diseases in which inflammation plays a role.The aim of this observational study was to assess RANTES levels in peripheral blood as clinical indicators of AD.Plasma levels of RANTES were investigated in 85 AD patients in a relatively early phase of AD(median 8.5 months after diagnosis;39 men and 46 women;average age 75.7 years),and in 78 control subjects(24 men and 54 women;average age 66 years).We found much higher plasma levels of RANTES in AD patients compared to controls.A negative correlation of RANTES levels with age,disease duration,Fazekas scale score,and the medial temporal lobe atrophy(MTA)score(Scheltens’s scale)was found in AD patients,i.e.,the higher levels corresponded to earlier stages of the disease.Plasma RANTES levels were not correlated with cognitive scores.In AD patients,RANTES levels were positively correlated with the levels of pro-inflammatory cytokines interleukin-6 and tumor necrosis factor-α,which is consistent with the wellknown fact that AD is associated with inflammatory processes.RANTES levels were also positively correlated with insulin levels in AD patients,with insulin resistance(HOMA-R)and pancreatic beta cell function(HOMA-F).This study evaluated several clinical and metabolic factors that may affect plasma levels of RANTES,but these factors could not explain the increases in RANTES levels observed in AD patients.Plasma levels of RANTES appear to be an interesting peripheral marker for early stages of AD.The study was approved by the Ethics Committee of Institute of Endocrinology,Prague,Czech Republic on July 22,2011.

Epithelial markers of colorectal carcinogenesis in ulcerative colitis and primary sclerosing cholangitis

AIM:To evaluate the expression of epithelial markers of colorectal carcinogenesis in patients with long-term ulcerative colitis(UC) and primary sclerosing cholangitis(PSC) before and after transplantation.METHODS:Eight patients with UC and PSC prior to liver transplantation(PSC-UC),22 patients with UC after liver transplantation for PSC(OLT),9 patients with active ulcerative colitis without PSC(UCA),7 patients withUC in remission(UCR) and 10 controls(N) underwent colonoscopy with multiple biopsies.Specimens were analysed histologically and semi-quantitatively immunohistochemically for p53,Bcl-2 and cyclooxygenase-2(COX-2) markers.Statistical analysis was performed by Kruskal-Wallis and Fisher's exact tests.RESULTS:PSC-UC had a statistically significantly higher expression of p53 in the nondysplastic mucosa as compared to OLT,UCA,UCR and N(P < 0.05).We also found a statistically significant positive correlation between the incidence of PSC and the expression of p53(P < 0.001).UCA had a higher p53 expression as compared to UCR.OLT had a significantly lower expression of p53 as compared with PSC-UC(P < 0.001).Bcl-2 had a significant higher bcl-2 expression as compared with controls.No difference in COX-2 expression between PSC-UC,UCR and UCA was found.UCA had higher COX-2 expression as compared to UCR.We also found a statistically significant positive correlation between the expression of COX-2 and p53.Patients after liver transplantation for PSC had a statistically significantly lower expression of the p53 compared with PSCUC(P < 0.001).PSC-UC had the same inflammatory endoscopic activity as OLT and UCR when evaluated with the Mayo score.CONCLUSION:Our study shows that the nondysplatic mucosa of UC patients with PSC is characterised by a higher expression of the tumour suppressor gene p53,suggesting a higher susceptibility of cancer.This p53 overexpression correlates with the presence of PSC whilst it is not present in patients with UC after liver transplantation for PSC.

Parallel fluorescence and phosphorescence monitoring of singlet oxygen photosensitization in rats

The time-and spectral-resolved set-up for measurements of weak infrared luminescence of photosensitizers(PSs)and singlet oxygen using optical lightguides was used on skin of laboratory animals in vivo.Wistar rats with abdominal incisions treated by methylis aminolevulinitis(MAL)were used as a model.A control group of animals with abdominal incisions was also tested.Spectrally resolved fluorescence of the PS was acquired during the treatment from the same spot.The intensity and spectral profile of the fiuorescence signal from the skin can be used to guide the detection setup to the investigated spots in the lesion.The rate of bleaching of Protoporphyrin IX band and appearance of a band of its photoproducts during the treatment can be characterized by the exposition ED,under which the latter becomes dominant feature infiuorescence spectrum.ED value differs statistically significantly between the normal skin and the lesion treated by MAL.No direct proportionality was found between the fluorescence signal and singlet oxygen production.Nevertheless,the strong fluorescence signal is necessary but not a su±cient condition for higher singlet oxygen production in vivo.ED value correlates rather well with production of singlet oxygen,but differently in lesion and normal skin.Lifetimes of singlet oxygen differ between spots outside and in the lesion.PS triplet state lifetimes exhibit weak di®erence between spots treated and untreated by MAL.

Visual Performance of a Polynomial Extended Depth of Focus Intraocular Lens

Purpose: To clinically evaluate a new extended depth of focus intraocular lens (ISOPURE, PhysIOL) with optic design modification based on a unique polynomial concept to improve intermediate vision while keeping the quality of distance vision equal to a monofocal lens. Methods: 18 patients (11 female, 7 male, mean age of 69.4 years) with bilateral cataract and regular corneal astigmatism ≤ 1.0 D underwent bilateral cataract surgery with ISOPURE implantation. Patients were followed for up to 6 months. Measured parameters were uncorrected (UDVA) and corrected distance visual acuity (CDVA), uncorrected (UIVA) and distance-corrected intermediate visual acuity at 80 cm and 66 cm (DCI80VA, DCI66VA) subjective refraction, defocus curve, tolerance of cylinder induction, and contrast sensitivity. The data from all implanted eyes (all-eyes) and a subset only including the first eye implanted for each patient were analysed. Results: The mean manifest refraction spherical equivalent (MRSE) decreased from 1.05 D pre-operatively to ?0.15 D at the 4 - 6 month assessment, with 80.6% of eyes within ±0.50 D of emmetropia. At the final follow-up, mean (SD) monocular CDVA was ?0.06 (0.04) logMAR, DCI80VA was 0.18 (0.08) logMAR and DCI66VA was 0.27 (0.13) logMAR. Despite a cylinder induction of ?0.50 D, uncorrected distance visual acuity of 0.02 logMAR was still achieved. Conclusion: The ISOPURE intraocular lens provides excellent distance corrected visual acuity for far and intermediate distances along with high contrast sensitivity and good tolerance of residual refractive cylinder.

New insights in bilirubin metabolism and their clinical implications

Bilirubin,a major end product of heme breakdown,is an important constituent of bile,responsible for its characteristic colour.Over recent decades,our understanding of bilirubin metabolism has expanded along with the processes of elimination of other endogenous and exogenous anionic substrates,mediated by the action of multiple transport systems at the sinusoidal and canalicular membrane of hepatocytes.Several inherited disorders characterised by impaired bilirubin conjugation(Crigler-Najjar syndrome typeⅠand typeⅡ,Gilbert syndrome)or transport(Dubin-Johnson and Rotor syndrome)result in various degrees of hyperbilirubinemia of either the predominantly unconjugated or predominantly conjugated type.Moreover,disrupted regulation of hepatobiliary transport systems can explain jaundice in many acquired liver disorders.In this review,we discuss the recent data on liver bilirubin handling based on the discovery of the molecular basis of Rotor syndrome.The data show that a substantial fraction of bilirubin conjugates is primarily secreted by MRP3 at the sinusoidal membrane into the blood,from where they are subsequently reuptaken by sinusoidal membrane-bound organic anion transporting polypeptides OATP1B1 and OATP1B3.OATP1B proteins are also responsible for liver clearance of bilirubin conjugated in splanchnic organs,such as the intestine and kidney,and for a number of endogenous compounds,xenobiotics and drugs.Absence of one or both OATP1B proteins thus may have serious impact on toxicity of commonly used drugs cleared by this system such as statins,sartans,methotrexate or rifampicin.The liverblood cycling of conjugated bilirubin is impaired in cholestatic and parenchymal liver diseases and this impairment most likely contributes to jaundice accompanying these disorders.

Dubin-Johnson syndrome coinciding with colon cancer and atherosclerosis

Hyperbilirubinemia has been presumed to prevent the process of atherogenesis and cancerogenesis mainly by decreasing oxidative stress.Dubin-Johnson syndrome is a rare,autosomal recessive,inherited disorder characterized by biphasic,predominantly conjugatedhyperbilirubinemia with no progression to end-stage liver disease.The molecular basis in Dubin-Johnson syndrome is absence or deficiency of human canalicular multispecific organic anion transporter MRP2/cMOAT caused by homozygous or compound heterozygous mutation(s) in ABCC2 located on chromosome 10q24.Clinical onset of the syndrome is most often seen in the late teens or early adulthood.In this report,we describe a case of previously unrecognized Dubin-Johnson syndrome caused by two novel pathogenic mutations (c.2360_2366delCCCTGTC and c.3258+1G>A),coinciding with cholestatic liver disease in an 82-year-old male patient.The patient,suffering from advanced atherosclerosis with serious involvement of coronary arteries,developed colorectal cancer with nodal metastases.The subsequent findings do not support the protective role of Dubin-Johnson type hyperbilirubinemia.

ABCB4 mutations underlie hormonal cholestasis but not pediatric idiopathic gallstones

AIM:To investigate the contribution of ABCB4 mutations to pediatric idiopathic gallstone disease and the potential of hormonal contraceptives to prompt clinical manifestations of multidrug resistance protein 3 deficiency.METHODS:Mutational analysis of ABCB4,screening for copy number variations by multiplex ligation-dependent probe amplification,genotyping for low expression allele c.1331T>C of ABCB11 and genotyping for variation c.55G>C in ABCG8 previously associated with cholesterol gallstones in adults was performed in 35 pediatric subjects with idiopathic gallstones who fulfilled the clinical criteria for low phospholipid-associated cholelithiasis syndrome(LPAC,OMIM#600803)and in 5young females with suspected LPAC and their families(5 probands,15 additional family members).The probands came to medical attention for contraceptiveassociated intrahepatic cholestasis.RESULTS:A possibly pathogenic variant of ABCB4was found only in one of the 35 pediatric subjects with idiopathic cholesterol gallstones whereas 15 members of the studied 5 LPAC kindreds were confirmed and another one was highly suspected to carry predictably pathogenic mutations in ABCB4.Among these 16,however,none developed gallstones in childhood.In 5index patients,all young females carrying at least one pathogenic mutation in one allele of ABCB4,manifestation of LPAC as intrahepatic cholestasis with elevated serum activity of gamma-glutamyltransferase was induced by hormonal contraceptives.Variants ABCB11c.1331T>C and ABCG8 c.55G>C were not significantly overrepresented in the 35 examined patients with suspect LPAC.CONCLUSION:Clinical criteria for LPAC syndrome caused by mutations in ABCB4 cannot be applied topediatric patients with idiopathic gallstones.Sexual immaturity even prevents manifestation of LPAC.

The immune hunger games:the effects of fasting on monocytes

Currently,different types of fasting are becoming increasingly popular for their potential health benefits,particularly in improving cardiometabolic diseases.However,how these practices affect immune function is not well understood.In a recent study published in Immunity,Janssen et al.delve into the complex relationship among fasting,refeeding,and the immune system.While fasting caused monocyte homing in bone marrow(BM),refeeding escalated monocyte counts in the circulation but altered immune responses to bacterial infection[1].

Interaction of Diet/Lifestyle Intervention and TCF7L2 Genotype on Glycemic Control and Adiposity among Overweight or Obese Adults:Big Data from Seven Randomized Controlled Trials Worldwide

Objective.The strongest locus which associated with type 2 diabetes(T2D)by the common variant rs7903146 is the transcription factor 7-like 2 gene(TCF7L2).We aimed to quantify the interaction of diet/lifestyle interventions and the genetic effect of TCF7L2 rs7903146 on glycemic traits,body weight,or waist circumference in overweight or obese adults in several randomized controlled trials(RCTs).Methods.From October 2016 to May 2018,a large collaborative analysis was performed by pooling individualparticipant data from 7 RCTs.These RCTs reported changes in glycemic control and adiposity of the variant rs7903146 after dietary/lifestyle-related interventions in overweight or obese adults.Gene treatment interaction models which used the genetic effect encoded by the allele dose and common covariates were applicable to individual participant data in all studies.Results.In the joint analysis,a total of 7 eligible RCTs were included(n=4,114).Importantly,we observed a significant effect modification of diet/lifestyle-related interventions on the TCF7L2 variant rs7903146 and changes in fasting glucose.Compared with the control group,diet/lifestyle interventions were related to lower fasting glucose by-3.06(95%CI,-5.77 to-0.36)mg/dL(test for heterogeneity and overall effect:I^(2)=45:1%,p<0:05;z=2:20,p=0:028)per one copy of the TCF7L2 T risk allele.Furthermore,regardless of genetic risk,diet/lifestyle interventions were associated with lower waist circumference.However,there was no significant change for diet/lifestyle interventions in other glycemic control and adiposity traits per one copy of TCF7L2 risk allele.Conclusions.Our findings suggest that carrying the TCF7L2 T risk allele may have a modestly greater benefit for specific diet/lifestyle interventions to improve the control of fasting glucose in overweight or obese adults.

四逆软肝方对高浓度乙醛培养的人肝星状细胞肝纤四项及MMP-1、TIMP-1的影响

目的:探讨四逆软肝方对高浓度乙醛培养的人肝星状细胞肝纤四项(CG、HA、LN、PCIII)及基质金属蛋白酶1(MMP-1)、组织金属蛋白酶抑制剂1(TIMP-1)的影响。方法:采用高浓度乙醛对人肝星状细胞LX-2株造成损伤后,随机分为四逆软肝方、清热解毒化浊片、扶正化瘀片和脱离乙醛4组;经给药24h后搜集各组细胞培养液上清,采用ELISA试剂盒检测肝纤四项及MMP-1、TIMP-1含量。结果:250mmol/L乙醛培养72h,细胞存活率最高;与正常组比,乙醛组CG、HA、LN、MMP-1、TIMP-1差异无统计学意义(P>0.05),PCIII升高(P<0.01)。与正常组比较,脱离乙醛组CG下降(P<0.01)、PCIII升高(P<0.01);与脱离乙醛组比较,四逆软肝方组HA、PCIII、TIMP-1均降低(P<0.01),CG、LN、MMP-1差异无统计学意义(P>0.05)。四逆软肝方组、清热解毒化浊片组、扶正化瘀片组组间比较,差异无统计学意义(P>0.05)。结论:四逆软肝方可降低受损的LX-2细胞HA、PCIII、TIMP-1指标的表达,可能对抗肝纤维化有一定的作用。