Fibrosis Grade Research Articles

Quantitative Analysis of Bone Marrow Features Highlights Heterogeneity in Myelofibrosis Patients Treated with Zinpentraxin Alfa in a Phase II Clinical Study

Quantitative image analysis has potential to transform the interpretation of bone marrow trephine (BMT) samples in myeloproliferative neoplasms (MPN) and improve the evaluation of anti-fibrotic therapies. To investigate the utility of recently developed algorithms evaluating reticulin fibrosis and megakaryocyte features in myelofibrosis (MF), we analysed samples from a multi-center, phase II study of zinpentraxin alfa (ZPN, PRM-151). ZPN is a recombinant form of human pentraxin-2 (PTX2) that has shown clinical activity as monotherapy and in combination with ruxolitinib (RUX) in a phase II trial in patients with Int-1/-2 or high risk MF (NCT01981850). In stage 2 of this study, patients ineligible for, intolerant of, or with an inadequate response to RUX were randomised to receive 0.3, 3.0, or 10.0 mg/kg ZPN on Days 1, 3, and 5 of Cycle 1, and every 4 weeks thereafter for up to 9 cycles. We demonstrate the potential of quantitative image analysis to augment and refine conventional expert histological assessment of MF. Reticulin and H+E-stained BMTs from three timepoints (screening, C4D1 and C9D29) were analysed from 50/97 patients enrolled. Manual assessment (MA) of marrow fibrosis and megakaryocyte features was performed by blinded, independent central review. Assessment of fibrosis using the Continuous Indexing of Fibrosis (CIF) score and analysis of megakaryocytes was performed by automated analyses ( Ryou H, Leukaemia, 2022). There was a moderate correlation between the automated average CIF score and manual fibrosis grade (Spearman's rho = 0.39). However, there was significant overlap in the distribution of CIF scores between samples manually assigned to MF-2 and MF-3. Approximately 38% of MF-2 samples fell within the interquartile range of CIF distribution observed in MF-3, and around 48% of MF-3 samples fell within the interquartile range observed in MF-2. Next, we visualised the screening BMTs in fibrosis PCA (principal component analysis) space to capture both average CIF score and Shannon entropy (‘unevenness‘) of CIF score for each sample. This revealed significant heterogeneity in reticulin fibrosis at screening when compared to an independent reference cohort of 130 MPN samples ( Ryou H, Leukaemia, 2022), likely reflecting the effects of pre-treatment [ Figure 1]. To track fibrosis across all timepoints we plotted the average CIF score for each sample. This revealed striking variability in fibrotic response within the trial cohort, with 15 of 42 patients (36%) demonstrating improvement in average CIF score [ Figure 2]. Notably, improvements in average CIF score by cycle 9 appeared to be most marked in patients with higher CIF scores at screening. No significant ZPN dose-dependent effect on fibrosis was observed. We next measured megakaryocyte density (megakaryocytes per unit area of intertrabecular space) and compared it to MA. We found a strong correlation (Spearman's rho = 0.81) between automated and MA of megakaryocyte density, with no significant change across any of the treatment arms. For megakaryocyte clustering (defined as ≥ 3 megakaryocytes in direct contact) we observed a moderate to strong correlation (Spearman's rho = 0.62) between MA and automated assessment, with no significant change across any of the treatment arms. Finally, we sought to compare the sample megakaryocyte cell features (cytomorphology and topology) at screening with those of an independent reference cohort of 88 MPN samples ( Sirinukunwattana, Blood Adv., 2020). Strikingly, not only was there marked heterogeneity in megakaryocyte features between patient samples at screening, but the observed features were significantly different to those of newly diagnosed MF. No significant changes in megakaryocyte cytomorphology or topology were seen across any of the treatment arms. In summary, our findings reveal considerable BMT morphological heterogeneity in patients participating in a Phase II trial of ZPN. This likely reflects poorly understood and under-recognised variation in morphological features encountered in patients with longstanding and/or pre-treated MF when compared to newly diagnosed patients. This highlights the potential of such variability to confound the evaluation of novel therapeutics in MPN, and emphasises the utility of robust quantitative methods to analyse and visualise morphological features in clinical study samples that can complement conventional manual assessment.

Application of Intravoxel Incoherent Motion in Clinical Liver Imaging: A Literature Review.

Intravoxel incoherent motion (IVIM) modeling is a widely used double-exponential model for describing diffusion-weighted imaging (DWI) signal, with a slow component related to pure molecular diffusion and a fast component associated with microcirculatory perfusion, which compensates for the limitations of traditional DWI. IVIM is a noninvasive technique for obtaining liver pathological information and characterizing liver lesions, and has potential applications in the initial diagnosis and treatment monitoring of liver diseases. Recent studies have demonstrated that IVIM-derived parameters are useful for evaluating liver lesions, including nonalcoholic fatty liver disease (NAFLD), liver fibrosis and liver tumors. However, the results are not stable. Therefore, it is necessary to summarize the current applications of IVIM in liver disease research, identify existing shortcomings, and point out the future development direction. In this review, we searched for studies related to hepatic IVIM-DWI applications over the past two decades in the PubMed database. We first introduce the fundamental principles and influential factors of IVIM, and then discuss its application in NAFLD, liver fibrosis, and focal hepatic lesions. It has been found that IVIM is still unstable in ensuring the robustness and reproducibility of measurements in the assessment of liver fibrosis grade and liver tumors differentiation, due to inconsistent and substantial overlap in the range of IVIM-derived parameters for different fibrotic stages. In the end, the future direction of IVIM-DWI in the assessment of liver diseases is discussed, emphasizing the need for further research on the stability of IVIM-derived parameters, particularly perfusion-related parameters, in order to promote the clinical practice of IVIM-DWI. LEVEL OF EVIDENCE: 5 TECHNICAL EFFICACY: Stage 3.

Histopathological Study of the Effects of Dura Adhesive Agents Used in Spinal Surgery Practice on Spinal Epidural Fibrosis in Experimental Animal Model.

Histopathological examination of the effects of Tisseel, Cova, Glubran and Coseal, which are used for sealing purposes in spinal surgery practice, on epidural fibrosis is aimed. Forty Sprague Dawley rats were randomly divided into five groups in our study as Group 1 (n=8) control group (Laminectomy); Group 2 (n=8) Cova group (Laminectomy + Cova); Group 3 (n=8) Tissel group (Laminectomy + Tisseel); Group 4 (n=8) Coseal group (Laminectomy + Coseal); and Group 5 Glubrane group (Laminectomy + Glubrane). Control group was only applied laminectomy. After laminectomy to other groups, Cova was applied to the 2nd group, Tissel to the 3rd group, Coseal to the 4th group and Glubran to the 5th group in surgical fields. After the rats were monitored in separate cages for 6 weeks after the operation, the relevant spinal level was extracted and the samples were examined histopathologically and the results were evaluated statistically. It was found that there was a statistically significant difference in Tisseel and Glubran groups in terms of fibrosis grading compared to the control group, and this had a positive effect on fibrosis. Compared to the control group, there was no statistically significant difference on fibrosis in Cova and Coseal groups. As dura adhesive agents used in spinal surgery practice did not increase spinal epidural fibrosis statistically significantly, we concluded that these products can be used safely during spinal surgery if necessary.

Point-of-care detection of fibrosis in liver transplant surgery using near-infrared spectroscopy and machine learning.

Visual assessment and imaging of the donor liver are inaccurate in predicting fibrosis and remain surrogates for histopathology. We demonstrate that 3-s scans using a handheld near-infrared-spectroscopy (NIRS) instrument can identify and quantify fibrosis in fresh human liver samples. We undertook NIRS scans on 107 samples from 27 patients, 88 from 23 patients with liver disease, and 19 from four organ donors. Liver disease patients had a median immature fibrosis of 40% (interquartile range [IQR] 20-60) and mature fibrosis of 30% (10%-50%) on histopathology. The organ donor livers had a median fibrosis (both mature and immature) of 10% (IQR 5%-15%). Using machine learning, this study detected presence of cirrhosis and METAVIR grade of fibrosis with a classification accuracy of 96.3% and 97.2%, precision of 96.3% and 97.0%, recall of 96.3% and 97.2%, specificity of 95.4% and 98.0% and area under receiver operator curve of 0.977 and 0.999, respectively. Using partial-least square regression machine learning, this study predicted the percentage of both immature (R 2 = 0.842) and mature (R 2 = 0.837) with a low margin of error (root mean square of error of 9.76% and 7.96%, respectively). This study demonstrates that a point-of-care NIRS instrument can accurately detect, quantify and classify liver fibrosis using machine learning.

C-kitpos cells in the human left atrial appendage

BackgroundSubpopulations of myocardial c-kitpos cells have the ability to stimulate regeneration in ischemic heart disease by paracrine effects. The left atrial appendage (LAA), which is easy accessible during cardiac surgery, may represent a perfect source for c-kitpos cell extraction for autologous cell therapies in the living human. So far, frequency and distribution of c-kitpos cells in LAA are unknown. MethodsLAAs of patients who underwent cardiac surgery due to coronary artery disease (coronary artery bypass graft, CABG), valvular heart disease or both and of two body donors were examined. Tissue was fixed in 4 % paraformaldehyde, embedded in paraffin, dissected in consecutive sections and stained for c-kitpos cells. In parallel, grade of fibrosis, amount of fat per section and cells positive for mast cell tryptase were examined. ResultsWe collected 27 LAAs (37.0 % female, mean left ventricular ejection fraction 50.4 %, 63.0 % persistent atrial fibrillation (AF)). Most of the patients underwent combined CABG and valve surgery (51.9 %). C-kitpos cells were detected in 3 different regions: A) Attached to the epicardial fat layer, B) close to vascular structures and C) between cardiomyocytes. C-kitpos cells ranged from 0.05 c-kitpos cells per mm2 to 67.5 c-kitpos cells per mm2. We found no association between number of c-kitpos cells and type of AF, amount of fibrosis or amount of fat. Up to 72 % of c-kitpos cells also showed a positive staining for mast cell tryptase. ConclusionC-kitpos cells are frequent in LAAs of cardiovascular patients with a rather homogenous distribution throughout the LAA. The LAA can therefore be considered as a source for extraction of a reasonable quantity of autologous cardiac progenitor cells in the living human patient.

Serum vitamin D is associated with ultrasound-defined hepatic fibrosis

BackgroundEvidences from population-based investigations on the exact relationship between vitamin D and the severity of liver fibrosis remain debated and conflicting. Here, we aim to explore the relationship between serum vitamin D and ultrasound-defined advanced hepatic fibrosis in the US participants with nonalcoholic fatty liver disease (NAFLD). MethodsIn the retrospective study, individuals with intact information on interesting variables from the 2017–2018 National Health and Nutrition Examination Survey (NHANES) were included. NAFLD was diagnosed on the basis of controlling attenuation parameter (CAP) value≥ 274 dB/m without causes of other chronic hepatic diseases. We identified advanced fibrosis grades (F2) by liver stiffness measurement (LSM) score of ≥ 8.2 kPa in NAFLD patients. The impact of elevated serum vitamin D on the prevalence of hepatic fibrosis was assessed by multivariate logistic regression models on the basis of the NHANES recommended weights. ResultsThe study involved 1624 subjects with NAFLD in total, and 305 (18.28 %, weighted%) of whom were diagnosed with advanced hepatic fibrosis according to the definition based on parameters obtained from vibration controlled transient elastography (VCTE). In the multivariate logistic regression analysis, serum vitamin D presented a negative relationship to hepatic fibrosis with lower odds in patients with hepatic steatosis after being adjusted for potential confounding factors (fully adjusted: OR=0.47, 95 % CI: 0.24–0.90, p = 0.034). Our subgroup analysis revealed that the inverse relationship was still existed in males (fully adjusted: OR = 0.34, 95 % CI: 0.17–0.70, p = 0.014), non-obese subjects (fully adjusted: OR = 0.20, 95 % CI: 0.04–0.89, p = 0.042) and participants below 60 years (fully adjusted: OR = 0.43, 95 % CI: 0.21–0.90, p = 0.033), whereas in models adjusted for the potential confounding factors, no statistically significant correlation was noted in females, obese subjects or subjects with age≥ 60 years. ConclusionsThis large population-based investigation indicated that elevated serum vitamin D reduced the onset of advanced fibrosis diagnosed by ultrasound in males, non-obese subjects and younger participants with NAFLD.

Combi-Elasto Evaluation of the Degree of Liver Fibrosis in Children with Cholestatic Liver Disease.

Cholestatic liver disease is a common liver disease in infants and young children. Liver fibrosis is a key factor affecting the prognosis, and liver transplantation is the only treatment option for liver cirrhosis. This study aimed to explore the efficacy of Combi-elasto for diagnosing liver fibrosis in children affected by cholestatic liver disease. A total of 64 children with S1-S4-grade liver fibrosis were enrolled. The general data, routine ultrasound, Combi-elasto, aspartate aminotransferase-to-platelet ratio index (APRI) and Fibrosis-4 (FIB-4) score were compared among children with different grades of liver fibrosis, and the efficacy of the above indexes for evaluating the degree of liver fibrosis was reported. There were remarkable differences in liver size, liver echogenicity, Young's modulus (E), fibrosis index (FI), activity index (AI) and FIB-4 score among the groups (all p < 0.05). E and liver echogenicity were the independent impact factors of liver fibrosis. The areas under the curve of E, APRI, FIB-4 score and the combined model (E+ liver echogenicity) in the evaluation of liver fibrosis were 0.84, 0.61, 0.66 and 0.90, respectively. Ultimately, we concluded that CE is an effective method to evaluate liver fibrosis in children with cholestatic liver disease.

PRİMER İMMÜN TROMBOSİTOPENİDE ARTMIŞ RETİKÜLER LİF DERECESİ

OBJECTIVE: Bone marrow fibrosis in primary immune thrombocytopenia (ITP) has become a centre of attention since cases with fibrosis were reported during trombopoeitin receptor agonist therapies but, there have been few studies evaluating the fibrosis status of the patients at diagnosis. The aim of the study was to evaluate the impact of marrow fibrosis on especially response to treatment and prognosis in patients with ITP. MATERIAL AND METHODS: Bone marrow reticulin fiber grade, haemoglobin, platelets, age, sex, co-morbidities of the patients, hepatitis and autoimmune markers on admission, response, remission status and duration of remission and treatments were recorded from medical files of the patients and each parameter was evaluated for an association with reticular fiber grade in 53 patients with ITP. RESULTS: 79.3% of patients had marrow reticulin content grade 1 or more. No significant correlations were found between bone marrow reticular fiber grade and total blood count at diagnosis, response times to the first, second- and third-line treatment, platelet counts after treatment and time between two treatment lines, age, gender, presence of comorbidity and antinuclear antibody positivity and response rate and time. There was a significant and positive correlation between platelet count at diagnosis and age (p&amp;lt;0.05) and, there was a statistically significant and negative correlation between white blood and neutrophil count at diagnosis and age (p&amp;lt;0.05) CONCLUSIONS: For the first time, higher grade of fibrosis was found in patients with ITP. Prospective studies with follow-up bone marrow biopsies are needed to validate the link between ITP and autoimmune fibrosis.

Glucocorticoid Replacement for Adrenal Insufficiency and the Development of Non-Alcoholic Fatty Liver Disease.

Glucocorticoid excess is a known risk factor for non-alcoholic fatty liver disease (NAFLD). Our objective was to analyse the impact of glucocorticoid replacement therapy on the development of NAFLD and NAFLD-related fibrosis and, therefore, on cardiovascular as well as hepatic morbidity in patients with adrenal insufficiency. Two hundred and fifteen individuals with primary (n = 111) or secondary (n = 104) adrenal insufficiency were investigated for hepatic steatosis and fibrosis using the fatty liver index (FLI), NAFLD fibrosis score (NAFLD-FS), Fibrosis-4 Index (FiB-4) plus sonographic transient elastography. Results were correlated with glucocorticoid doses and cardiometabolic risk parameters. The median dose of hydrocortisone equivalent was 20 mg daily, with a median therapy duration of 15 years. The presence and grade of hepatic steatosis and fibrosis were significantly correlated with cardiometabolic risk factors. We could not find any significant correlations between single, daily or cumulative doses of glucocorticoids and the grade of liver steatosis, nor with fibrosis measured via validated sonographic techniques. In patients with adrenal insufficiency, glucocorticoid replacement within a physiological range of 15-25 mg hydrocortisone equivalent per day does not appear to pose an additional risk for the development of NAFLD, subsequent liver fibrosis, or the cardiovascular morbidity associated with these conditions.

Prospective quantitative study: Doppler ultrasound in the evaluation of chronic renal allograft disease and correlation with histopathological finding.

Renal allograft biopsy is the gold standard for diagnosing chronic allograft nephropathy, but noninvasive methods are needed to avoid unnecessary biopsies. Doppler ultrasonography, particularly the resistive index (RI), correlates with renal allograft dysfunction. This study aims to assess the relationship between renal sonographic parameters and biochemical parameters in diagnosing graft interstitial fibrosis. The study evaluated 60 renal allograft recipients for sonographic renal morphological features and Doppler indices. The estimated glomerular filtration rate (eGFR) was calculated, and cortical fibrosis after the biopsy was determined using the Banff score. Continuous variables like mean and SD were calculated, and categorical variables were reported using frequencies and proportions. Associations were examined using independent sample t-tests, χ 2tests, and multivariate regression analysis. The mean eGFR was 75.23±25.45ml/min/1.73m2. A significant correlation of eGFR with RI (r=0.341, P=0.008) was seen. A significant difference in mean RI (F=10.167; df=2,57; P<0.001) was seen among the histological grades of fibrosis. Among the histological grades of fibrosis, significant differences in RI among mild and moderate (S.E. 0.033, P=0.043), mild and severe (S.E. 0.026, P=0.001) as well as moderate and severe (S.E. 0.036, P=0.029) was seen. Doppler was able to noninvasively predict allograft fibrosis and could be used as a complementary imaging tool during the follow-up of renal allograft patients. Future research is needed to improve evidence, diagnostic criteria, guidelines, and long-term impact.

Drivers and high-molecular-risk mutations in Argentine patients with primary myelofibrosis

Primary myelofibrosis (PMF), a BCR-ABL negative myeloproliferative neoplasm, is a heterogeneous clinical and genetic disorder with a poor prognosis. We aimed to study the mutational profile of driver genes (JAK2, CALR, and MPL) and high-molecular-risk (HMR) genes (ASXL1, EZH2, IDH1/2, and SRSF2) and their prognostic impact in 65 Argentine patients with PMF. Mutually exclusive driver mutations were identified in 88% of the cases and HMR mutations were detected in 37% of patients, including 37.5% of triple negative cases. ASXL1 and SRSF2 were the most frequently mutated HMR genes, and a significant association between these mutations was observed (P = 0.04). Additionally, age &gt;65 years, lower hemoglobin level, constitutional symptoms, male sex, and bone marrow (BM) fibrosis grade ≥ 2 were correlated with ASXL1 mutations, and circulating blasts with SRSF2 mutations. HMR mutations were significantly associated with inferior overall survival (P &lt; 0.001). These mutations were more frequent with increasing risk of DIPSS score and allowed us to identify patients with poor survival in intermediate DIPSS groups. The application of the MIPSS70 score stratified our patients into three risk groups, with significant differences in overall survival (P &lt; 0.001). Interestingly, 85% of the patients who were upgraded to a higher risk category compared to the DIPSS score showed a HMR profile. In conclusion, our study contributes to the understanding of the mutational landscape in MFP, supports the integration of molecular data to improve prognostic models and reinforces the adverse prognostic impact of HMR mutations, particularly in ASXL1 and SRSF2 genes.

The Importance of 1-Year Protocol Biopsy in the Long-Term Prognosis of Kidney Transplants—5-Years Follow-Up

Protocol biopsies are performed to detect subclinical pathologies that may lead to future graft dysfunction. However, they are not routinely performed interventions in every transplant center. There is no established regimen for performing them. The study aimed to evaluate if protocol biopsies can improve long-term patient outcomes after detecting early disorders and modifying treatment. Our observational study included 61 patients who underwent protocol biopsy 12 months after the transplantation. Based on the biopsy results, patients with abnormal histologic material (n=37) were divided into 3 study groups as follows: patients with mild inflammatory lesions (n=21), patients with interstitial fibrosis and tubular atrophy (IFTA) grade II to III (n=12), and patients with BK virus nephropathy (n=4). The control group (n=24) included kidney recipients with IFTA 0 to I grade. Outcomes after 5-year follow-up were evaluated. Five years after the biopsy, patients in the control group had stable graft function (5-year change in serum creatinine was -0.09 mg/dL). An increase in serum creatinine levels was observed in patients with IFTA II to III compared with the control group (0.14 mg/dL, P=.04). Immunosuppressive treatment was modified in the group with mild inflammatory changes and in the BKV group after the biopsy result. In the group with mild inflammatory lesions, renal function was stable (change of serum creatinine was -0.01 mg/dL, P=.51). In the BKV nephropathy group, there was a significant reduction in serum creatine levels (-0.48 mg/dL, P=.016). The analysis showed no diagnostic value for serum creatinine concentration (95% CI 0.49-0.78, P=.08). Protocol biopsies are useful for detecting early pathologies and preventing allograft failure. They greatly benefit patients with detectable pathology that can be treated or in whom therapy modification is possible.

Meta-Analysis of Five Fraction Preoperative Radiotherapy for Soft Tissue Sarcoma

There is increasing interest in shorter courses of radiation therapy (RT) in the management of soft tissue sarcoma (STS). Studies investigating preoperative ultra-hypofractionated 5-fraction RT for STS are few and often limited to single institution experiences with less than 50 patients. We therefore performed a meta-analysis to determine the efficacy and safety of preoperative 5-fraction RT for STS based on currently published literature, with an analysis evaluating effects of dose delivered. This study level meta-analysis was conducted using Bayesian methods. Statistical estimation for risk of outcome rates were conducted by posterior mean and 95% highest posterior density (HPD) intervals. Studies with two-year local control (LC) and description of major wound complications (MWC) per the CAN-NCIC-SR2 study were included in this meta-analysis, and these served as the primary endpoints. Secondary endpoints included rates of acute and late toxicity. A total of ten studies were identified; seven met inclusion criteria (Kalbasi 2020, Kubicek 2021, Leite 2021, Spalek 2021, Gobo Silva 2021, Bedi 2022, Mayo 2022). Three studies (Kosela-Paterczyk 2014, Kosela-Paterczyk 2021, Potkrajcic 2021) did not include adequate information to accurately determine 2-year LC or MWC. Subgroup analyses were performed for ≥30 Gy vs <30 Gy as the EQD2 of 30 Gy in 5 fractions for an α/β of 3-4 is 50-54 Gy, equivalent to standard preoperative RT fractionation. A total of 208 patients were included from seven studies. Five studies used ≥30 Gy (n = 144) and 2 studies <30 Gy (n = 64). The median follow-up was 29 months (range: 21-57). Primary tumor location was lower extremity in 68%, upper extremity in 22%, and trunk in 10%. In studies with available information, most tumors were intermediate (27%, 46/169) or high grade (67%, 114/169) and 50% (79/158) were >10 cm. Two-year LC for the entire cohort was 96.9% (95% HPD: 0.9374-0.9889) and the rate of MWC was 30.6% (95% HPD: 0.2106-0.4149). Acute grade 2 and 3 dermatitis was seen in 12.7% and 2.2%, respectively. Late grade 2 toxicities included fibrosis (11.1%), stiffness (6.0%), and lymphedema (4.0%) and late grade 3 toxicities included fibrosis (1.8%) and stiffness (0.4%). Grade 4 toxicity was rare and included fibrosis (0.6%) and stiffness (0.6%). There was a trend toward improved LC with ≥ 30 Gy (95% HPD: 0.949-0.997 vs 0.838-0.986). There was no difference in MWC (95% HPD: 0.185-0.420 vs 0.172-0.553) or late toxicity between the two groups. Acute grade 3 dermatitis was seen less frequently with regimens <30 Gy (95% HPD: 0-0 vs 0.009-0.072), however rare overall. Preoperative 5-fraction RT for STS demonstrates excellent 2-year LC with MWC and toxicity similar to standard fractionation preoperative RT. Multi-institutional trials with a universal RT protocol and larger sample size are warranted to assess this novel treatment paradigm.

Distinct Morphological and Molecular Profiles of NAFLD and NAFLD-associated HCC Revealed by Immunohistochemistry and MicroRNA Analysis.

Non-alcoholic fatty liver disease (NAFLD) is a common hepatic condition that can progress to hepatocellular carcinoma (HCC) in non-cirrhotic livers. To better understand the development of NAFLD-associated HCC, we performed an integrated morphological and molecular analysis to identify new insights that can improve the follow-up of NAFLD patients. Our study included a cohort of 14 NAFLD-associated HCC and 41 NAFLD patients. We analyzed clinical parameters, a four-microRNA (miRNA) panel (miR-21-5p, miR-34a-5p, miR-130a-3p, and miR-155-3p) panel and their relationship with p53 and β-catenin expression. In the study cohort, the NAFLD-associated HCC patients were predominantly male, older, had significantly altered hepatic function, and a higher incidence of hypertension, type 2 diabetes, and dyslipidemia. Morphologically, the NAFLD-HCC group had substantially higher steatosis, ballooning, and fibrosis grades than the NAFLD group. The β-catenin expression was higher in both adjacent non-tumoral liver tissue (ANT) from NAFLD-associated HCC patients and in HCC tissue com-pared with NAFLD samples. The 4 miRNAs panel showed a dysregulated expression profile between NAFLD, ANT and HCC samples. This study provides important insights regarding the molecular mechanisms underlying HCC progression in NAFLD patients, allowing for the development of better screening strategies for the early detection of NAFLD-associated HCC.

Interleukin-34 is more suitable than macrophage colony-stimulating factor for predicting liver significant fibrosis in patients with chronic hepatitis B

Aims Interleukin-34 (IL-34) and macrophage colony-stimulating factor (CSF-1) have similar functions, such as promoting the formation of liver fibrosis. This study aimed to evaluate and compare the diagnostic value of serum IL-34 and CSF-1 for significant liver fibrosis in patients with chronic hepatitis B (CHB). Methods A total of 369 CHB patients, consisting of 208 HBeAg-negative patients and 161 HBeAg-positive patients, were enrolled in this study. Additionally, 72 healthy individuals served as healthy controls (HCs). Serum levels of IL-34 and CSF-1 were measured using the enzyme-linked immunosorbent assay method. Liver fibrosis grades were assessed using the modified Scheuer scoring system. Results Serum IL-34 and CSF-1 levels exhibited significant elevation in both HBeAg-negative and HBeAg-positive patients in comparison to HCs (p < 0.001). IL-34 emerged as an independent factor linked to significant liver fibrosis, whereas CSF-1 did not exhibit such an association. Receiver operating characteristic (ROC) analysis indicated higher areas under the curves (AUCs) for IL-34 (0.814, p < 0.001 and 0.673, p < 0.001) when diagnosing significant liver fibrosis in HBeAg-negative and HBeAg-positive patients, respectively, as opposed to CSF-1 (0.602, p < 0.001; 0.619, p = 0.385). Within the HBeAg-negative patient subgroup, the AUC for IL-34 surpassed that of FIB-4 (p = 0.009) and APRI (p = 0.045). Conclusion Serum IL-34 has the potential to be a straightforward and practical biomarker that demonstrates superior performance to serum CSF-1 in the diagnosis of significant liver fibrosis in CHB patients, especially within the HBeAg-negative patients.

Contouring variation affects estimates of normal tissue complication probability for breast fibrosis after radiotherapy

BackgroundNormal tissue complication probability (NTCP) models can be useful to estimate the risk of fibrosis after breast-conserving surgery (BCS) and radiotherapy (RT) to the breast. However, they are subject to uncertainties. We present the impact of contouring variation on the prediction of fibrosis. Materials and methods280 breast cancer patients treated BCS-RT were included. Nine Clinical Target Volume (CTV) contours were created for each patient: i) CTV_crop (reference), cropped 5 mm from the skin and ii) CTV_skin, uncropped and including the skin, iii) segmenting the 95% isodose (Iso95%) and iv) 3 different auto-contouring atlases generating uncropped and cropped contours (Atlas_skin/Atlas_crop). To illustrate the impact of contour variation on NTCP estimates, we applied two equations predicting fibrosis grade ≥ 2 at 5 years, based on Lyman-Kutcher-Burman (LKB) and Relative Seriality (RS) models, respectively, to each contour. Differences were evaluated using repeated-measures ANOVA. For completeness, the association between observed fibrosis events and NTCP estimates was also evaluated using logistic regression. ResultsThere were minimal differences between contours when the same contouring approach was followed (cropped and uncropped). CTV_skin and Atlas_skin contours had lower NTCP estimates (−3.92%, IQR 4.00, p < 0.05) compared to CTV_crop. No significant difference was observed for Atlas_crop and Iso95% contours compared to CTV_crop. For the whole cohort, NTCP estimates varied between 5.3% and 49.5% (LKB) or 2.2% and 49.6% (RS) depending on the choice of contours. NTCP estimates for individual patients varied by up to a factor of 4. Estimates from “skin” contours showed higher agreement with observed events. ConclusionContour variations can lead to significantly different NTCP estimates for breast fibrosis, highlighting the importance of standardising breast contours before developing and/or applying NTCP models.

Saturated Fat-Mediated Upregulation of IL-32 and CCL20 in Hepatocytes Contributes to Higher Expression of These Fibrosis-Driving Molecules in MASLD.

Metabolic dysfunction-associated steatotic liver disease (MASLD) comprises a spectrum of liver diseases, ranging from liver steatosis to metabolic dysfunction-associated steatohepatitis (MASH), increasing the risk of developing cirrhosis and hepatocellular carcinoma (HCC). Fibrosis within MASLD is critical for disease development; therefore, the identification of fibrosis-driving factors is indispensable. We analyzed the expression of interleukin 32 (IL-32) and chemokine CC ligand 20 (CCL20), which are known to be linked with inflammation and fibrosis, and for their expression in MASLD and hepatoma cells. RT-PCR, ELISA and Western blotting analyses were performed in both human liver samples and an in vitro steatosis model. IL-32 and CCL20 mRNA expression was increased in tissues of patients with NASH compared to normal liver tissue. Stratification for patatin-like phospholipase domain-containing protein 3 (PNPLA3) status revealed significance for IL-32 only in patients with I148M (rs738409, CG/GG) carrier status. Furthermore, a positive correlation was observed between IL-32 expression and steatosis grade, and between IL-32 as well as CCL20 expression and fibrosis grade. Treatment with the saturated fatty acid palmitic acid (PA) induced mRNA and protein expression of IL-32 and CCL20 in hepatoma cells. This induction was mitigated by the substitution of PA with monounsaturated oleic acid (OA), suggesting the involvement of oxidative stress. Consequently, analysis of stress-induced signaling pathways showed the activation of Erk1/2 and p38 MAPK, which led to an enhanced expression of IL-32 and CCL20. In conclusion, cellular stress in liver epithelial cells induced by PA enhances the expression of IL-32 and CCL20, both known to trigger inflammation and fibrosis.

Fibro-scan Based Score and Novel Noninvasive Serum Marker, FGF-21, to Assess Liver Fibrosis in Morbidly Obese Patients with Nonalcoholic Fatty Liver Disease

Abstract Background Non-alcoholic fatty liver disease (NAFLD) is currently the most common cause of liver disease in the Western world, affecting up to 20-30% of the general adult population. It is a growing public health problem, because of the increasing prevalence of the pathologies that contribute to its development such as obesity and inflammation status. Aim of the Work This prospective cohort study aims to develop and validate noninvasive scoring system for assessment liver fibrosis in morbidly obese patients with NAFLD. Subjects and Methods This is a Prospective cohort study, was carried out on 50 morbidly obese patients selected from endocrinology department, AIN SHAMS UNIVERSITY, through a period of six months. Results The main results of the study revealed that: There were 32(64.0%) male and 18(36.0%) female with mean age 40.48 (± 12.16) SD. There were 14(28.0%) between (20 -30), 11(22.0%) between (30-40), 9(18.0%) between (40-50) and 16(32.0%) ≥50. Mean Weight (Kg) was 106.6 (± 13.31) SD with range (82.50 – 145.5), mean Height (Cm) was 171.7 (± 6.25) SD with range (158.0 – 185.0), mean BMI (Kg/m2) was 36.17 (± 4.15) SD with range (29.30 – 43.40), mean Waist circumference 113.1 (± 8.64) SD with range (92.0 – 129.0). There were 37(74.0%) with Comorbidity. 10(20.0%) have DM, 11(22.0%) have HTN, 11(22.0%) have CDK and 5(10.0%) have COPD. mean Hb was 12.24 with range (11.0 – 13.50), mean Plts was 193.2 with range (151.0 – 250.0), mean WBCs with range (4.70 – 8.20), mean AST was 85.20 with range (21.0 – 210.0), mean ALT was 86.64 with range (15.0 – 212.0), mean Cholesterol was 181.4 with range (122.0 – 248.0), mean Triglycerides was 156.3 with range (12.0 – 294.0), mean LDL was 119.1 with range (61.0 – 193.0), mean HDL was 61.32 with range (23.0 – 96.0), mean GGT (U/l) was 18.22 with range (7.0 – 33.0), mean HBA1C was 5.72 with range (4.70 – 9.30), mean Fasting blood sugar was 101.8 with range (81.0 – 155.0) and mean Uric acid was 6.23 with range (4.0 – 10.10). Regarding ultrasound outcome there were 33(66.0%) have Gallbladder wall blurring, 34(68.0%) have Portal vein wall blurring, 42(84.0%) have Hepatic vein blurring and 50 (100.0%) have fatty liver. Regarding fibrosis grade there were 12(24.0%) F1, 17(34.0%) F2, 18(36.0%) F3 and 3(6.0%) F4. Conclusion That the combination of Fibroscan and, APRI, FGF-21 and FIB-4 methods provides a valuable approach for assessing liver fibrosis in NAFLD patients. This can eliminate the need for liver biopsy in patients without clear indication. In addition, several recent studies have also validated the used of non-invasive markers in the diagnosis of NAFLD. The establishment of a national program for the recognition of NAFLD is therefore essential to reduce the risk of liver disease progression.

Phytosomal curcumin and shilajit decrease adhesion bands post-Achilles tendon surgery in animal model

Aim: This study aimed to investigate the protective effects of phytosomal curcumin and/or shilajit in reducing post-surgical tendon adhesion band formation in a rat model. Background: Tendon adhesion is one of the severe complications after tendon surgery which causes limited tendon movement and functional disability. Objective: According to the central role of inflammatory reactions in fibrosis and the formation of tendon adhesions, we investigated the therapeutic effects of phytosomal curcumin and shilajit either alone or in combination on reducing post-surgical tendon adhesion band formation in a rat model. Methods: We randomly divided 36 Wistar male rats into six equal groups. (A) Control group with no surgical incision and no intervention. (B) Sham group with surgical incision, but no adhesion, (C) Positive control group with total surgical transection and adhesion receiving normal saline daily, (D-F) Treatment groups which are the same as group C except that rats were treated with the following drugs. Phytosomal curcumin alone (250 µL/day; orally), shilajit alone (500 mg/kg/day; orally) and a combination of phytosomal curcumin + shilajit. In groups with surgical intervention, the rats' Achilles tendons were cut and repaired with a modified Kessler technique. At 3 weeks, all rats were euthanized. Histological and pathological scoring systems were used to evaluate the protective effects of phytosomal curcumin and/or shilajit in reducing adhesion bands at the site of tendon injuries. Results: Our results revealed that the administration of phytosomal curcumin and/or shilajit remarkably reduced length, density, grading, severity, and thickness of post-surgical adhesion bands. Compared to the untreated control group, the histological changes and inflammatory reactions were significantly attenuated in treated rats. Furthermore, treatment with phytosomal curcumin and/or shilajit inhibited fibrotic responses by alleviating collagen deposition, fibrosis quantity, fibrosis grading, and total fibrosis scores, as visualized by Masson’s trichrome staining. Conclusion: Our findings indicated the anti-inflammatory and anti-fibrotic properties of phytosomal curcumin and/or shilajit supporting their therapeutic potential in preventing post-operative tendon adhesion bands.

Liver fibrosis screening, evaluation pathway, and management in patients with fatty liver

Non-alcoholic fatty liver disease (NAFLD) is not a benign condition, especially in patients with non-alcoholic steatohepatitis (NASH) combined with liver fibrosis grades F2-4, who have a higher risk of liver-related events and mortality. Thus, this population is considered "at-risk" for developing NASH. China has a large NAFLD patient population, so how to screen for those with liver fibrosis is an important socio-economic concern. At the moment, serological models, liver stiffness detection based on vibration-controlled transient elastography, and magnetic resonance elastography (MRE) are the only non-invasive tests (NITs) for liver fibrosis. The prevention and treatment guidelines for NAFLD at home and abroad are reviewed here, based on the research progress of NITs in recent years, so as to suggest screening, evaluation pathways, and management for liver fibrosis in patients with NAFLD.