The concept of gene therapy for glaucoma:the dream that has not come true yet

Gene therapies,despite of being a relatively new therapeutic approach,have a potential to become an important alternative to current treatment strategies in glaucoma.Since glaucoma is not considered a single gene disease,the identified goals of gene therapy would be rather to provide neuroprotection of retinal ganglion cells,especially,in intraocular-pressure-independent manner.The most commonly reported type of vector for gene delivery in glaucoma studies is adeno-associated virus serotype 2 that has a high tro pism to retinal ganglion cells,res ulting in long-term expression and low immunogenic profile.The gene thera py studies recruit inducible and genetic animal models of optic neuropathy,like DBA/2J mice model of high-tension glaucoma and the optic nerve crush-model.Reported gene therapy-based neuroprotection of retinal ganglion cells is targeting specific genes translating to growth factors(i.e.,brain derived neurotrophic factor,and its receptor TrkB),regulation of apoptosis and neurodegeneration(i.e.,Bcl-xl,Xiap,FAS system,nicotinamide mononucleotide adenylyl transferase 2,Digit3 and Sarm1),immunomodulation(i.e.,Crry,C3 complement),modulation of neuroinflammation(i.e.,e rythropoietin),reduction of excitotoxicity(i.e.,Com KIlα)and transcription regulation(i.e.,Max,Nrf2).On the other hand,some of gene therapy studies focus on lowering intra ocular pressure,by impacting genes involved in both,decreasing aqueous humor production(i.e.,aquaporin 1),and increasing outflow facility(i.e.,COX2,prostaglandin F2a receptor,RhoA/RhoA kinase signaling pathway,MMP1,Myocilin).The goal of this review is to summarize the current stateof-art and the direction of development of gene therapy strategies for glaucomatous neuropathy.

Bone graft incorporation failure with inappropriate limb load transfer can lead to aseptic acetabular loosening of metal-on-metal prosthesis: A case report

BACKGROUND Aseptic acetabular loosening can result from various factors that can be categorized into groups:patient-related,surgeon-related and implant-related.We present a case of a 63-year-old patient who at first underwent a total hip arthro-plasty(THA)using a metal-on-metal bearing due to hip arthrosis.Follow-up visits revealed no complications after the procedure.Two years after the THA,acetabular component loosening occurred due to subsequent trauma of the opposite hip,necessitating a revision THA using a ceramic-on-ceramic bearing.CASE SUMMARY We aim to illustrate a rare case where the primary reason for undergoing THA revision was not only incomplete bone graft incorporation but also improper limb load distribution.Following the revision arthroplasty,a 9-year follow-up visit revealed improvements in all evaluation measures on questionnaire compared to the state before surgery:Harris Hip Score(before surgery:15;after surgery:95),Western Ontario and McMaster Universities Arthritis Index(before surgery:96;after surgery:0),and Visual Analogue Scale(before surgery:10;after surgery:1).CONCLUSION Opposite-hip trauma caused a weight transfer to the limb after a THA procedure.This process led to a stress shielding effect,resulting in acetabular component loosening.

Evaluation of the nutritional status of patients with liver cirrhosis

BACKGROUND Progressive malnutrition coexists with liver diseases,particularly in patients with cirrhosis.Early diagnosis of malnutrition in patients with advanced stages of chronic liver disease and the implementation of appropriate nutritional treatment for malnourished patients should be an integral part of the therapeutic process.AIM To evaluate the nutritional status of patients with various severities of advanced liver fibrosis,using various nutritional status parameters.METHODS This study involved 118 patients with liver cirrhosis who were classified into three groups according to their Child-Pugh score.The nutritional status of the patients in each group was assessed using different methods.The average values obtained from the measurements were calculated for each research group.The influence of disease stage on the examined parameters of nutritional status was determined using one-way analysis of variance.To investigate the relationship between the parameters determining nutritional status and the stage of disease advancement,a correlation analysis was performed.RESULTS The Child-Pugh A group had the highest mean body weight(76.42 kg),highest mean body mass index(BMI)(26.72 kg/m²),and largest mean arm circumference(27.64 cm).In the Child-Pugh B group,the mean scores of all examined variables were lower than those of the Child-Pugh A group,whereas the mean body weight and BMI of the Child-Pugh C group were higher than those of the Child-Pugh B group.There was a very strong correlation between the Child-Pugh classification and subjective global assessment score;a very strong correlation between the Child-Pugh classification and arm circumference;a strong correlation between the Child-Pugh classification and body weight,albumin concentration,fat-free mass index,muscle mass index,phase angle,and BMI;and an average correlation between Child-Pugh classification and fat mass index.Notably,these indicators deteriorated with disease progression.CONCLUSION Advanced liver fibrosis leads to the deterioration of many nutritional status parameters.The extent of malnutrition increases with the progression of liver fibrosis.The Child-Pugh score reflects the nutritional status.

Real-world effectiveness and safety of direct-acting antivirals in hepatitis C virus patients with mental disorders

BACKGROUND It is estimated that 58 million people worldwide are infected with the hepatitis C virus(HCV).Patients with severe psychiatric disorders could not be treated with previously available interferon-based therapies due to their unfavorable side effect profile.This has changed with the introduction of direct-acting antivirals(DAA),although their real-life tolerance and effectiveness in patients with different psychiatric disorders remain to be demonstrated.AIM To evaluate the effectiveness and safety of DAA in patients with various mental illnesses.METHODS This was a retrospective observational study encompassing 14272 patients treated with DAA for chronic hepatitis C in 22 Polish hepatology centers,including 942 individuals diagnosed with a mental disorder(anxiety disorder,bipolar affective disorder,depression,anxiety-depressive disorder,personality disorder,schizophrenia,sleep disorder,substance abuse disorder,and mental illness without a specific diagnosis).The safety and effectiveness of DAA in this group were compared to those in a group without psychiatric illness(n=13330).Antiviral therapy was considered successful if serum ribonucleic acid(RNA)of HCV was undetectable 12 wk after its completion[sustained virologic response(SVR)].Safety data,including the incidence of adverse events(AEs),serious AEs(SAEs),and deaths,and the frequency of treatment modification and discontinuation,were collected during therapy and up to 12 wk after treatment completion.The entire study population was included in the intent-to-treat(ITT)analysis.Per-protocol(PP)analysis concerned patients who underwent HCV RNA evaluation 12 wk after completing treatment.RESULTS Among patients with mental illness,there was a significantly higher percentage of men,treatmentnaive patients,obese,human immunodeficiency virus and hepatitis B virus-coinfected,patients with cirrhosis,and those infected with genotype 3(GT3)while infection with GT1b was more frequent in the population without psychiatric disorders.The cure rate calculated PP was not significantly different in the two groups analyzed,with a SVR of 96.9% and 97.7%,respectively.Although patients with bipolar disorder achieved a significantly lower SVR,the multivariate analysis excluded it as an independent predictor of treatment non-response.Male sex,GT3 infection,cirrhosis,and failure of previous therapy were identified as independent negative predictors.The percentage of patients who completed the planned therapy did not differ between groups with and without mental disorders.In six patients,symptoms of mental illness(depression,schizophrenia)worsened,of which two discontinued treatments for this reason.New episodes of sleep disorders occurred significantly more often in patients with mental disorders.Patients with mental illness were more frequently lost to follow-up(4.2%vs 2.5%).CONCLUSION DAA treatment is safe and effective in HCV-infected patients with mental disorders.No specific psychiatric diagnosis lowered the chance of successful antiviral treatment.

Changes in characteristics of patients with hepatitis C virus-related cirrhosis from the beginning of the interferon-free era

BACKGROUND Nearly 290000 patients with chronic hepatitis C die annually from the most severe complications of the disease.One of them is liver cirrhosis,which occurs in about 20%of patients chronically infected with the hepatitis C virus(HCV).Direct-acting antivirals(DAAs),which replaced interferon(IFN)-based regimens,significantly improved the prognosis of this group of patients,increasing HCV eradication rates and tolerability of therapy.Our study is the first to assess changes in patient profile,effectiveness,and safety in the HCV-infected cirrhotic population in the IFN-free era.AIM To document changes in patient characteristics and treatment regimens along with their effectiveness and safety profile over the years.METHODS The studied patients were selected from 14801 chronically HCV-infected individuals who started IFN-free therapy between July 2015 and December 2021 in 22 Polish hepatology centers.The retrospective analysis was conducted in real-world clinical practice based on the EpiTer-2 multicenter database.The measure of treatment effectiveness was the percentage of sustained virologic response(SVR)calculated after excluding patients lost to follow-up.Safety data collected during therapy and the 12-wk post-treatment period included information on adverse events,including serious ones,deaths,and treatment course.RESULTS The studied population(n=3577)was balanced in terms of gender in 2015-2017,while the following years showed the dominance of men.The decline in the median age from 63 in 2015-2016 to 61 years in 2021 was accompanied by a decrease in the percentage of patients with comorbidities and comedications.Treatment-experienced patients dominated in 2015-2016,while treatment-naive individuals gained an advantage in 2017 and reached 93.2%in 2021.Genotype(GT)-specific options were more prevalent in treatment in 2015-2018 and were supplanted by pangenotypic combinations in subsequent years.The effectiveness of the therapy was comparable regardless of the period analyzed,and patients achieved an overall response rate of 95%,with an SVR range of 72.9%-100%for the different therapeutic regimens.Male gender,GT3 infection,and prior treatment failure were identified as independent negative predictors of therapeutic success.CONCLUSION We have documented changes in the profile of HCV-infected cirrhotic patients over the years of accessibility to changing DAA regimens,confirming the high effectiveness of IFN-free therapy in all analyzed periods.

Metabolic dysfunction-associated steatotic liver disease:Navigating terminological evolution,diagnostic frontiers and therapeutic horizon-an editorial exploration

Metabolic dysfunction-associated steatotic liver disease(MASLD),once known as non-alcoholic fatty liver disease(NAFLD),represents a spectrum of liver disorders characterized by lipid accumulation within hepatocytes.The redefinition of NAFLD in 2023 marked a significant reposition in terminology,emphasizing a broader understanding of liver steatosis and its associated risks.MASLD is now recognized as a major risk factor for liver cirrhosis,hepatocellular carcinoma,and systemic complications such as cardiovascular diseases or systemic inflammation.Diagnostic challenges arise,particularly in identifying MASLD in lean individuals,necessitating updated diagnostic protocols and investing in non-invasive diagnostic tools.Therapeutically,there is an urgent need for effective treatments targeting MASLD,with emerging pharmacological options focusing on,among others,carbohydrate and lipid metabolism.Additionally,understanding the roles of bile acid metabolism,the microbiome,and dietary interventions in MASLD pathogenesis and management holds promise for innovative therapeutic approaches.There is a strong need to emphasize the importance of collaborative efforts in understanding,diagnosing,and managing MASLD to improve physicians’approaches and patient outcomes.

Nodular regenerative hyperplasia:Evolving concepts on underdiagnosed cause of portal hypertension

Nodular regenerative hyperplasia(NRH)is a rare liver condition characterized by a widespread benign transformation of the hepatic parenchyma into small regenerative nodules.NRH may lead to the development of non-cirrhotic portal hypertension.There are no published systematic population studies on NRH and our current knowledge is limited to case reports and case series.NRH may develop via autoimmune,hematological,infectious,neoplastic,or drug-related causes.The disease is usually asymptomatic,slowly or nonprogressive unless complications of portal hypertension develop.Accurate diagnosis is made by histopathology,which demonstrates diffuse micronodular transformation without fibrous septa.Lack of perinuclear collagen tissue distinguishes NRH from typical regenerative nodules in the cirrhotic liver.While the initial treatment is to address the underlying disease,ultimately the therapy is directed to the management of portal hypertension.The prognosis of NRH depends on both the severity of the underlying illness and the prevention of secondary complications of portal hypertension.In this review we detail the epidemiology,pathogenesis,diagnosis,management,and prognosis of NRH.

Effect of interferon alpha and ribavirin treatment on serum levels of transforming growth factor-β1,vascular endothelial growth factor,and basic fibroblast growth factor in patients with chronic hepatitis C

AIM： To assess the role of transforming growth factor-β1 （TGF-β1）, vascular endothelial growth factor （VEGF） and basic fibroblast growth factor （bFGF） in the pathogenesis of fibrosis associated with chronic hepatitis C （CHC） and to evaluate the influence of the antiviral therapy on above parameter levels depending on the treatment results （complete response or no response）. METHODS： Study group included 100 patients with CHC, in whom fibrosis in liver specimens was assessed （Scheuer fibrosis score： 1-4 points）. Control group included 30 subjects with antibodies anti-HCV present and persistently normal ALT level, without fibrosis （Scheuer fibrosis score： 0 points）. Concentration of studied parameters was assayed in the serum by immunoenzymatic method before and after the therapy with interferon alpha-2b and ribavirin. RESULTS： TGF-β1 levels were significantly higher in the study group compared to the control group （35.89 vs 32.37 ng/mL; P= 0.023）. Such differences were not found in VEGF and bFGF levels. In patients showing complete response （negative HCV RNA and normal ALT level）, significant increase in VEGF （112.8 vs 315.03 pg/mL; P〈 0.05） and bFGF （2.51 vs 15.79 pg/mL; P=0.04） levels were found. Significant decrease in TGF-β1 level was observed both in responders （37.44 vs 30.02 ng/mL; P=0.05）, and in non-responders （38.22 vs 30.43 ng/mL; P=0.043）. bFGF levels before the treatment were significantly lower （2.51 vs 5.94 pg/mL; P=0.04）, and after the treatment significantly higher （15.79 vs 4.35 pg/mL; P=0.01） in patients with complete response response. CONCLUSION： Among the analyzed parameters TGF-β1 seems to play the most important role in the pathogenesis of fibrosis in CHC. Levels of this factor are significantly lower in subjects who do not have fibrosis developed in them. Good therapeutic effect in CHC patients is associated with significant changes in TGF-β1, VEGF, and bFGF levels, bFGF seems to have the highest usefulness in the prognosis of treatment efficacy.

Intestinal epithelium, intraepithelial lymphocytes and the gut microbiota-Key players in the pathogenesis of celiac disease

Celiac disease(CD) is a chronic immune-mediated disorder triggered by the ingestion of gluten in genetically predisposed individuals. Before activating the immune system, gluten peptides are transferred by the epithelial barrier to the mucosal lamina propria, where they are deamidated by intestinal tissue transglutaminase 2. As a result, they strongly bind to human leucocyte antigens(HLAs), especially HLA-DQ2 and HLA-DQ8, expressed on antigen-presenting cells. This induces an inflammatory response, which results in small bowel enteropathy. Although gluten is the main external trigger activating both innate and adaptive(specific) immunity, its presence in the intestinal lumen does not fully explain CD pathogenesis. It has been hypothesized that an early disruption of the gut barrier in genetically susceptible individuals, which would result in an increased intestinal permeability, could precede the onset of gluten-induced immune events. The intestinal barrier is a complex functional structure, whose functioning is dependent on intestinal microbiotahomeostasis, epithelial layer integrity, and the gutassociated lymphoid tissue with its intraepithelial lymphocytes(IELs). The aim of this paper was to review the current literature and summarize the role of the gut microbiota, epithelial cells and their intercellular junctions, and IELs in CD development.

Impact of sodium glucose cotransporter-2 inhibitors on liver steatosis/fibrosis/inflammation and redox balance in non-alcoholic fatty liver disease

BACKGROUND Sodium glucose cotransporter-2 inhibitors(SGLT2-I)are the most recently approved drugs for type 2 diabetes(T2D).Recent clinical trials of these compounds reported beneficial cardiovascular(CV)and renal outcomes.A major cause of vascular dysfunction and CV disease in diabetes is hyperglycemia associated with inflammation and oxidative stress.Pre-clinical studies demonstrated that SGLT2-I reduce glucotoxicity and promote anti-inflammatory effects by lowering oxidative stress.AIM To investigate the effects of SGLT2-I on markers of oxidative stress,inflammation,liver steatosis,and fibrosis in patients of T2D with non-alcoholic fatty liver disease(NAFLD).METHODS We referred fifty-two consecutive outpatients treated with metformin monotherapy and exhibiting poor glycemic control to our centre.We introduced the outpatients to an SGLT2-I(dapagliflozin,empagliflozin,or canagliflozin;n=26)or a different hypoglycemic drug[other glucose-lowering drugs(OTHER),n=26].We evaluated circulating interleukins and serum hydroxynonenal(HNE)-or malondialdehyde(MDA)-protein adducts,fatty liver index(FLI),NAFLD fibrosis score,aspartate aminotransferase(AST)/alanine aminotransferase(ALT)ratio,AST-to-platelet-ratio index(APRI),and fibrosis-4 on the day before(T0)and following treatment for six months(T1).We also performed transient elastography at T0 and T1.RESULTS Add-on therapy resulted in improved glycemic control and reduced fasting blood glucose in both groups.Of note,following treatment for six months,a reduction of FLI and APRI,as well as of the FibroScan result,was reported in patients treated with SGLT2-I,but not in the OTHER group;furthermore,in the SGLT2-I group,we reported lower circulating levels of interleukin(IL)-1β,IL-6,tumor necrosis factor,vascular endothelial growth factor,and monocyte chemoattractant protein-1,and higher levels of IL-4 and IL-10.We did not observe any modification in circulating interleukins in the OTHER group.Finally,serum HNE-and MDA-protein adducts decreased significantly in SGLT2-I rather than OTHER patients and correlated with liver steatosis and fibrosis scores.CONCLUSION The present data indicate that treatment with SGLT2-I in patients with T2D and NAFLD is associated with improvement of liver steatosis and fibrosis markers and circulating pro-inflammatory and redox status,more than optimizing glycemic control.

Usefulness of two independent hist classifications of tumor regression iUsefulness of two independent histopathological classifications of tumor regression in patients with rectal cancer submitted to hyperfractionated pre-operative radiotherapy

AIM： To assess the usefulness of two independent histopathological classifications of rectal cancer regression following neo-adjuvant therapy. METHODS： Forty patients at the initial stage cT3NxM0 submitted to preoperative radiotherapy （42 Gy during 18 d） and then to radical surgical treatment. The relationship between ＂T-downstaging＂ versus regressive changes expressed by tumor regression grade （TRG 1-5） and Nasierowska-Guttmejer classification （NG 1-3） was studied as well as the relationship between TRG and NG versus local tumor stage ypT and lymph nodes status, ypN. RESULTS： Complete regression （ypT0, TRG 1） was found in one patient. ＂T-downstaging＂ was observed in 11 （27.5%） patients. There was a weak statistical significance of the relationship between ＂T-downstaging＂ and TRG staging and NG stage. Patients with ypT1 were diagnosed as TRG 2-3 while those with ypT3 as TRGS. No lymph node metastases were found in patients with TRG 1-2. None of the patients without lymph node metastases were diagnosed as TRG 5. Patients in the ypT1 stage were NG 1-2. No lymph node metastases were found in NG 1. There was a significant correlation between TRG and NG. CONCLUSION： Histopathological classifications may be useful in the monitoring of the effects of hyperfractionated preoperative radiotherapy in patients with rectal cancer at the stage of cT3NxM0. There is no unequivocal relationship between ＂Todownstaging＂ and TRG and NG. There is some concordance in the assessment of lymph node status with ypT, TRG and NG. TRG and NG are of limited value for the risk assessment of the lymph node involvement.

Short- and medium-term reproducibility of gastric emptying of a solid meal determined by a low dose of ^(13)C-octanoic acid and nondispersive isotope-selective infrared spectrometry

AIM： To evaluate the reproducibility of a modified 13^C breath test-based measurement of solid phase gastric emptying （GE） within the frames of a simple-repeated measure study protocol. METHODS： Twelve healthy subjects （6 females and 6 males, mean age 24.9＋0.7 years） were recruited to undergo three identical GE examinations. In six subjects the first two examinations were performed 2 d apart, and the third session was carried out at a median interval of 19.5 d （range 18 - 20 d） from the second one. In another six subjects the first two measurements were taken 20 d apart （median, range： 17-23 d）, whereas the third session took place 2 d after the second one. Probes of expiratory air collected before and during six hours after intake of a solid meal （378 kcal） labelled with 75 μL （68 mg） 13^C-octanoic acid, were measured for 13^CO2 enrichment with the nondispersive isotopeselective infrared spectrometry NDIRS apparatus. RESULTS： Taking coefficients of variation for paired examinations into account, the short-term reproducibility of the GE measurement was slightly but not significantly better than the medium-term one： 7.7% and 11.2% for the lag phase （T-Lag）, 7.3% and 10.9% for the gastric half emptying time （T1/2）. The least differences in GE parameters detectable at P= 0.05 level in the 12 paired examinations were 9.6 and 15.6 min for T-Lag, 11.6 and 19.7 min for T1/2 by a two-day or two to three-week time gap, respectively CONCLUSION： The low-cost modification of the breath test involving a lower dose of 13^C-octanoic acid and NDIRS, renders good short- and medium-term reproducibility, as well as sensitivity of the measurement of gastric emptying of solids.

Simultaneous liver mucinous cystic and intraductal papillary mucinous neoplasms of the bile duct:A case report

Cystic hepatic neoplasms are rare tumors,and are classified into two separate entities:mucinous cystic neoplasms(MCNs)and intraductal papillary mucinous neoplasms of the bile duct(IPMN-B).We report the case of a 56-year-old woman who presented with abdominal pain and jaundice due to the presence of a large hepatic multilocular cystic tumor associated with an intraductal tumor.Partial hepatectomy with resection of extrahepatic bile ducts demonstrated an intrahepatic MCN and an intraductal IPMN-B.This is the first report of the simultaneous occurrence of these two histologically distinct entities in the liver.

Use of ebselen as a neuroprotective agent in rat spinal cord subjected to traumatic injury

Spinal cord injury(SCI) causes disturbances of motor skills.Free radicals have been shown to be essential for the development of spinal cord trauma.Despite some progress,until now no effective pharmacological therapies against SCI have been verified.The purpose of our experiment was to investigate the neuroprotective effects of ebselen on experimental SCI.Twenty-two rats subjected to SCI were randomly subjected to SCI with no further treatment(n = 10) or intragastric administration of ebselen(10 mg/kg) immediately and 24 hours after SCI.Behavioral changes were assessed using the Basso,Beattie,and Bresnahan locomotor scale and footprint test during 12 weeks after SCI.Histopathological and immunohistochemical analyses of spinal cords and brains were performed at 12 weeks after SCI.Magnetic resonance imaging analysis of spinal cords was also performed at 12 weeks after SCI.Rats treated with ebselen presented only limited neurobehavioral progress as well as reduced spinal cord injuries compared with the control group,namely length of lesions(cysts/scars)visualized histopathologically in the spinal cord sections was less but cavity area was very similar.The same pattern was found in T2-weighted magnetic resonance images(cavities) and diffusion-weighted images(scars).The number of FluoroGold retrogradely labeled neurons in brain stem and motor cortex was several-fold higher in ebselen-treated rats than in the control group.The findings suggest that ebselen has only limited neuroprotective effects on injured spinal cord.All exprimental procedures were approved by the Local Animal Ethics Committee for Experiments on Animals in Katowice(Katowice,Poland)(approval No.19/2009).

Is endoscopic therapy the treatment of choice in all patients with chronic pancreatitis?

Chronic pancreatitis(CP) is a progressive inflammatory disease of the pancreas characterized by destruction of the pancreatic parenchyma with subsequent fibrosis that leads to pancreatic exocrine and endocrine insufficiency.Abdominal pain and local complications(bile duct or duodenal stenosis and pancreatic tumor) secondary to CP are indications for therapy.At the beginning,medical therapy is used.More invasive treatment is recommended for patients with pancreatic duct stones(PDS) and pancreatic obstruction in whom standard medical therapy is not sufficient.Recently,Clarke et al assessed the long-term effectiveness of endoscopic therapy(ET) in CP patients.The authors compared ET with medical treatment.They reported that ET was clinically successful in 50% of patients with symptomatic CP.In this commentary,current CP treatment,including indications for ET and surgery in CP patients,is discussed.Recommendations for endoscopic treatment of CP according to the European Society of Gastrointestinal Endoscopy Clinical Guidelines are reviewed.Different surgical methods used in the treatment of CP patients are also discussed.ET is the most useful in patients with large PDS,pancreatic duct obstruction and dilation.It should be the first-line option because it is less invasive than surgery.Surgery should be the first-line option in patients in whom ET has failed or in those with a pancreatic mass with suspicion of malignancy.ET is a very effective and less invasive procedure,but it cannot be recommended as the treatment of choice in all CP patients.

Impact of senescence on the transdifferentiation process of human hepatic progenitor-like cells

BACKGROUND Senescence is characterized by a decline in hepatocyte function,with impairment of metabolism and regenerative capacity.Several models that duplicate liver functions in vitro are essential tools for studying drug metabolism,liver diseases,and organ regeneration.The human HepaRG cell line represents an effective model for the study of liver metabolism and hepatic progenitors.However,the impact of senescence on HepaRG cells is not yet known.AIM To characterize the effects of senescence on the transdifferentiation capacity and mitochondrial metabolism of human HepaRG cells.METHODS We compared the transdifferentiation capacity of cells over 10(passage 10[P10])vs P20.Aging was evaluated by senescence-associated(SA)beta-galactosidase activity and the comet assay.HepaRG transdifferentiation was analyzed by confocal microscopy and flow cytometry(expression of cluster of differentiation 49a[CD49a],CD49f,CD184,epithelial cell adhesion molecule[EpCAM],and cytokeratin 19[CK19]),quantitative PCR analysis(expression of albumin,cytochrome P4503A4[CYP3A4],γ-glutamyl transpeptidase[γ-GT],and carcinoembryonic antigen[CEA]),and functional analyses(albumin secretion,CYP3A4,andγ-GT).Mitochondrial respiration and the ATP and nicotinamide adenine dinucleotide(NAD^(+))/NAD with hydrogen(NADH)content were also measured.RESULTS SAβ-galactosidase staining was higher in P20 than P10 HepaRG cells;in parallel,the comet assay showed consistent DNA damage in P20 HepaRG cells.With respect to P10,P20 HepaRG cells exhibited a reduction of CD49a,CD49f,CD184,EpCAM,and CK19 after the induction of transdifferentiation.Furthermore,lower gene expression of albumin,CYP3A4,andγ-GT,as well as reduced albumin secretion capacity,CYP3A4,andγ-GT activity were reported in transdifferentiated P20 compared to P10 cells.By contrast,the gene expression level of CEA was not reduced by transdifferentiation in P20 cells.Of note,both cellular and mitochondrial oxygen consumption was lower in P20 than in P10 transdifferentiated cells.Finally,both ATP and NAD^(+)/NADH were depleted in P20 cells with respect to P10 cells.CONCLUSION SA mitochondrial dysfunction may limit the transdifferentiation potential of HepaRG cells,with consequent impairment of metabolic and regenerative properties,which may alter applications in basic studies.

Impact of ultrasound and optical biometry on refractive outcomes of cataract surgery after penetrating keratoplasty in keratoconus

AIM: To analyse the impact of ultrasound and optical intraocular lens(IOL) calculation methods on refractive outcomes of cataract phacoemulsification performed after penetrating keratoplasty(PK) in keratoconus. METHODS: Phacoemulsification cataract surgery was performed on 42 eyes of 34 patients with keratoconus who had previously undergone PK. The IOL power was determined by using both standard and corneal topography-derived keratometry using the SRK/T formula. We used two independent methods-ultrasound biometry(UB) and interferometry [optical biometry(OB)] for IOL calculation. The analysed data from medical records included demographics, medical history, best corrected visual acuity(BCVA) on Snellen charts, technique of IOL calculation and calculation formula and its impact on final refractive result.RESULTS: BCVA ranged from 0.01 to 0.4(mean 0.09±0.19) before surgery and ranged from 0.2 to 0.7(mean 0.38±0.14) at 1 mo and from 0.2 to 1.0(mean 0.56±0.16)(P<0.05) at 3 mo, postoperatively. The refractive aim differed significantly from the refractive outcome in both the UB and OB groups(P<0.05). There was no statistically significant difference in the accuracy of the two biometry methods.CONCLUSION: The refractive aim in keratoconus eyes post-PK is not achieved with either ultrasound or OB.

Ascites as a Symptom of a Rotavirus Infection—Case Report

Rotaviruses (RV) are the most common cause of acute diarrhoea in children. The course of RV in-fections may vary from a mild to a serious course of diarrhoea with significant disturbances to the water-electrolyte and acid-base homeostasis and complications requiring hospitalisation. There are known cases of postinfectious enteropathy and severe cases may involve a complete villous atrophy and temporary intolerance to lactose in the course of rotavirus infection. Opinions on the changes in the permeability of the mucous of the small intestine in the result of ongoing inflammation are largely diverse. This study presents a very rare complication of a rotavirus infection which is ascites in a 3-month infant. It seems that the pathomechanism of ascites in presented patient is very complex and affected by other factors such as nutritional disorders, including hypoproteinemia and hypoalbuminemia and immunological factors.

Composite neuroendocrine and adenomatous carcinoma of the papilla of Vater

Malignant tumors of papilla are usually adenocarcinomas. We present a 67-year-old female who became icteric as result of a malignant tumor infiltrating the papilla of Vater. Histopathological assessment of surgically excised tumor showed both neuroendocrine and adenocarcinomatous features. To our knowledge, this is the seventh report of this rare neoplastic association in the duodenal periampullary region.

Cell therapy of a patient with type Ⅲ Osteogenesis imperfecta caused by mutation in COL1A2 gene and unstable collagen type I

The allogenic bone marrow derived mesenchymal stem cells transplantation was given to the newborn girl diagnosed with osteogenesis imperfecta type III, with multiple bone fractures, extreme shortness and limbs deformities. The treatment was performed at the age of 4 and 6 weeks. The clinical diagnosis was supported by biochemical analysis of collagen type I recovered from culture medium of cultivated patient’s skin fibroblast, which revealed its triple helix instability at temperature about 2?C lower than normal. Sequencing of both genes encoding procollagen type I revealed heterozygous substitution G23569Ain COL1A2 gene causing change of glycine at position 517 to aspartate. The donor of mesenchymal stem cells was the girl’s father. She received two intravenous infusions of suspended cultured mesenchymal cells in 16 days apart without any side effects. An analysis of procollagen type I secreted to the culture medium by bone marrow-derived mesenchymal stem cells obtained from the patient, 3 months following transplantation revealed its normal triple helix stability. During the subsequent two years of follow up two new bone fractures were noted. Currently a two-year-old girl’s presents extreme growth and weight deficiency. The motoric development is also retarded, but the patient constantly improves and makes progresses.