Collagen Fibrosis Research Articles

P4 LEFT ATRIAL APPENDAGE ANATOMICAL CHANGES FOLLOWING RADIOFREQUENCY–BASED OSTIAL ISOLATION

Abstract Background Left atrial appendage (LAA) electrical isolation (ei) may be achieved via radiofrequency (RF) energy applications at the level of the appendage ostium targeting the sites of earliest activation recorded by a mapping catheter. Notably, RF has long been used in vascular, orthopedic, and aesthetic surgery to promote thermal–induced collagen matrix contraction, fibrosis, and tissue retraction. LAA anatomical changes associated to RF–induced tissue retraction have never been reported. Objective To quantify the anatomical changes of the LAA ostium following RF–based LAAei. Methods Thirty–four consecutive patients requiring AF ablation with LAAei underwent transesophageal echocardiography (TEE) within 7 days before (baseline TEE) and >6 months after (follow–up TEE) ablation. The diameter of LAA orifice and landing zone were measured at 4 different views (0°, 45°, 90°, 135°). Measurements were performed by two independent reviewers blinded to the patient’s identity. Results Among 34 AF patients (68±7yrs, 73.5% males), the LAA morphology was classified as chicken wing in 15 (44%) patients, windsock in 10 (29%), cactus in 6 (18%), and cauliflower in 3 (9%). At baseline TEE, the mean maximum and mean minimum ostial diameters were 25±4mm and 22±4mm, respectively. The mean maximum and mean minimum diameters of the landing zone were 26±4mm and 23±3mm, respectively. On average, LAAei was achieved after 16±7 minutes of RF at a power of 45–47W. Follow–up TEE was performed 257±148 days after LAAei. The median LAA contraction velocity was 0.1 m/s (IQR: 0.04–0.18) and was significantly impaired in all patients. At follow–up TEE, the mean maximum and mean minimum ostial diameters were 19±4mm and 17±3mm, respectively. The mean maximum and mean minimum diameters of the landing zone were 20±4mm and 18±4mm, respectively. The mean relative reduction of the ostium and the landing zone were –24.4% and –22.5%, respectively. Box–Whisker plots of the maximum and minimum ostial diameters before and after LAAei are reported in Fig. 1. Conclusion RF led to a > 20% reduction of the diameters of the ostium and the landing zone. These changes may have important implications for a successful percutaneous occlusion procedure and justify a staged approach of isolation and occlusion.

FGF19 Is Downregulated in Idiopathic Pulmonary Fibrosis and Inhibits Lung Fibrosis in Mice.

Idiopathic pulmonary fibrosis (IPF) is a devastating lung disease with limited therapeutic possibilities. FGF19 (fibroblast growth factor 19), an endocrine FGF, was recently shown to decrease liver fibrosis. To ask whether FGF19 had antifibrotic properties in the lung and decipher its effects on common features associated with lung fibrogenesis, we assessed, by ELISA, FGF19 concentrations in plasma and BAL fluids obtained from control subjects and patients with IPF. In vivo, using an intravenously administered adeno11-associated virus, we overexpressed FGF19 at the fibrotic phase of two experimental models of murine lung fibrosis and assessed its effect on lung morphology, lung collagen content, fibrosis markers, and profibrotic mediator expression at mRNA and protein levels. In vitro, we investigated whether FGF19 could modulate the TGF-β-induced differentiation of primary human lung fibroblasts into myofibroblasts and the apoptosis of murine alveolar type II cells. Although FGF19 was not detected in BAL fluid, FGF19 concentration was decreased in the plasma of patients with IPF compared with control subjects. In vivo, the overexpression of FGF19 was associated with a marked decrease of lung fibrosis and fibrosis markers, with a decrease of profibrotic mediator expression and lung collagen content. In vitro, FGF19 decreased alveolar type 2 epithelial cell apoptosis through the decrease of the proapoptotic BIM protein expression and prevented TGF-β-induced myofibroblast differentiation through the inhibition of JNK phosphorylation. Altogether, these data identify FGF19 as an antifibrotic molecule with potential therapeutic interest in fibrotic lung disorders.

D-Carvone Attenuates CCl4-Induced Liver Fibrosis in Rats by Inhibiting Oxidative Stress and TGF-ß 1/SMAD3 Signaling Pathway.

Simple SummaryLiver fibrosis is a challenging global health problem resulting in a significant morbidity and mortality rates worldwide due to its rapid progression to cirrhosis and hepatocellular carcinoma. Therefore, identifying nontoxic therapies with precise curative effects to slow the progression of liver fibrosis comprises one of the most popular and high-priority areas of current research. D-carvone is a naturally occurring monoterpene abundant in the essential oil of aromatic plants such as caraway and spearmint. In the present study, the protective impact of D-carvone on carbon tetrachloride (CCl4)-induced liver fibrosis in rats was evaluated. Administration of D-carvone significantly enhanced liver functions, oxidant/antioxidant balance as well as liver histology. D-carvone ameliorated the progression of liver fibrosis, evident by the decreased collagen deposition (fibrosis score) and the reduced expression of the pro-fibrogenic markers TGF-β1 and SMAD3 in the liver. These findings reveal the anti-fibrotic effects of D-carvone and suggest that D-carvone could be a promising candidate for therapeutic intervention of liver fibrosis and other oxidative stress-related hepatic diseases.D-carvone is a natural monoterpene found in abundance in the essential oil of aromatic medicinal plants with a wide range of pharmacological values. However, the impact of D-carvone on liver fibrosis remains unclear. This study aimed to evaluate the anti-fibrotic potential of D-carvone in a rat model of liver fibrosis and to clarify the possible underlying mechanisms. Liver fibrosis was induced in rats by carbon tetrachloride, CCl4 (2.5 mL/kg, interperitoneally every 72 h for 8 weeks). Oral treatment of rats with D-carvone (50 mg/kg, daily) started on the 3rd week of CCl4 administration. D-carvone significantly enhanced liver functions (ALT, AST), oxidant/antioxidant status (MDA, SOD, GSH, total antioxidant capacity; TAC), as well as histopathological changes. Moreover, D-carvone effectively attenuated the progression of liver fibrosis, evident by the decreased collagen deposition and fibrosis score by Masson trichrome staining (MT) and α-SMA protein expression. Moreover, D-carvone administration resulted in a significant downregulation of the pro-fibrogenic markers TGF-β1 and SMAD3 and upregulation of MMP9. These findings reveal the anti-fibrotic effect of D-carvone and suggest regulation of the TGF-β1/SMAD3 pathway, together with the antioxidant activity as a mechanistic cassette, underlines this effect. Therefore, D-carvone could be a viable candidate for inhibiting liver fibrosis and other oxidative stress-related hepatic diseases. Clinical studies to support our hypothesis are warranted.

Pirfenidone ameliorates chronic pancreatitis in mouse models through immune and cytokine modulation

Chronic pancreatitis (CP) is an irreversible fibro-inflammatory disease of the pancreas with no current targeted therapy. Pirfenidone, an anti-fibrotic and anti-inflammatory drug, is FDA approved for treatment of Idiopathic Pulmonary Fibrosis (IPF). Its efficacy in ameliorating CP has never been evaluated before. We recently reported that pirfenidone improves acute pancreatitis in mouse models. The aim of the current study was to evaluate the therapeutic efficacy of pirfenidone in mouse models of CP. We used caerulein and L-arginine models of CP and administered pirfenidone with ongoing injury, or in well-established disease. We evaluated for fibrosis by Sirius-red staining for collagen, immunohistochemistry, western blotting, and qPCR for fibrosis markers to show the salutary effects of pirfenidone in CP. Our results suggest that treatment with pirfenidone ameliorated CP related changes in the pancreas (i.e., atrophy, acinar cell loss, fibrosis, and inflammation) not only when administered with ongoing injury, but also in well-established models of caerulein as well as L-arginine induced CP. It reduces the pro-fibrotic phenotype of macrophages (in-vivo and in-vitro), reduces macrophage infiltration into the pancreas and alters the intra-pancreatic cytokine milieu preceding changes in histology. The therapeutic effect of pirfenidone is abrogated in absence of macrophages. Furthermore, it reduces collagen secretion, cytokine levels and fibrosis markers in pancreatic stellate cells in-vitro. As it is FDA approved, our findings in mouse models simulating clinical presentation of patients to the clinic, can be used as the basis of a clinical trial evaluating the efficacy of this drug as a therapeutic agent for CP.

PO-696-07 LEFT ATRIAL APPENDAGE ANATOMICAL CHANGES FOLLOWING RADIOFREQUENCY-BASED OSTIAL ISOLATION

Left atrial appendage (LAA) electrical isolation (ei) may be achieved via radiofrequency (RF) energy applications at the level of the appendage ostium targeting the sites of earliest activation recorded by a mapping catheter. Notably, RF has long been used in vascular, orthopedic, and aesthetic surgery to promote thermal-induced collagen matrix contraction, fibrosis, and tissue retraction. LAA anatomical changes associated to RF-induced tissue retraction have never been reported. To quantify the anatomical changes of the LAA ostium following RF-based LAAei. Thirty-four consecutive patients requiring AF ablation with LAAei underwent transesophageal echocardiography (TEE) within 7 days before (baseline TEE) and >6 months after (follow-up TEE) ablation. The diameter of LAA orifice and landing zone were measured at 4 different views (0°, 45°, 90°, 135°). Measurements were performed by two independent reviewers blinded to the patient’s identity. Among 34 AF patients (68±7yrs, 73.5% males), the LAA morphology was classified as chicken wing in 15 (44%) patients, windsock in 10 (29%), cactus in 6 (18%), and cauliflower in 3 (9%). At baseline TEE, the mean maximum and mean minimum ostial diameters were 25±4mm and 22±4mm, respectively. The mean maximum and mean minimum diameters of the landing zone were 26±4mm and 23±3mm, respectively. On average, LAAei was achieved after 16±7 minutes of RF at a power of 45-47W. Follow-up TEE was performed 257±148 days after LAAei. The median LAA contraction velocity was 0.1 m/s (IQR: 0.04-0.18) and was significantly impaired in all patients. At follow-up TEE, the mean maximum and mean minimum ostial diameters were 19±4mm and 17±3mm, respectively. The mean maximum and mean minimum diameters of the landing zone were 20±4mm and 18±4mm, respectively. The mean relative reduction of the ostium and the landing zone was -24.4% and -22.5%, respectively. Box-Whisker plots of the maximum and minimum ostial diameters before and after LAAei are reported in Fig.1. RF led to a >20% reduction of the diameters of the ostium and the landing zone. These changes may have important implications for a successful percutaneous occlusion procedure and justify a staged approach of isolation and occlusion.

Mutation in the 5'SL Region of Collagen I mRNA Attenuates Diastolic Dysfunction in Male, but not Female, Mice Following 28 Weeks of Feeding a High Fat High Sucrose Diet

IntroductionThe La ribonucleoprotein 6 (LARP6), an RNA binding protein, increases type I collagen expression by regulating mRNA stability and translation. This is mediated via its interaction with the 5’ stem loop (5’SL) structure in collagen I mRNA. Disrupting this interaction by mutating the 5’SL region has been shown to reduce fibrotic remodeling in liver. Therefore, we tested the hypothesis that this interaction plays a role in cardiac fibrosis and stiffening in diet induced obesity in 5’SL mutant and wild‐type littermate mice. In fact, obesity and hypertension promote remodeling and cardiac stiffening due to excess collagen deposition, fibrosis, and microvascular dysfunction, that ultimately lead to diastolic dysfunction.MethodsMale and female 5’SL mice and wild‐type (WT) littermates were fed a high fat high sucrose (HFHS) or control diet starting at 6 weeks of age. After 28 weeks of feeding, mice were assessed for cardiac function by echocardiography. Hearts were collected and a transverse section of the left and right ventricle was fixed for histology. RNA was isolated from the left ventricle and submitted for mRNA sequencing and qPCR.ResultsFollowing 28 weeks of a HFHS diet, male and female mice showed increased body weight, cardiac hypertrophy (heart weight normalized to tibia length), and diastolic dysfunction. However, the 5’SL mutation attenuated the impaired diastolic function in male, but not female mice. Surprisingly, cardiac fibrosis was not altered by the HFHS diet. RNA sequencing revealed that HFHS diet altered gene expression associated with glucose and ketone metabolism, as well as inflammation, in male WT littermates. However, the 5’SL mutation enriched gene networks associated with vascular development and angiogenesis.ConclusionsChronic HFHS feeding leads to increased body weight, hypertrophic cardiomyopathy, and diastolic dysfunction in both male and female mice. However, the 5’SL mutation blunts dysfunction only in male mice, demonstrating sex‐dependent cardioprotective effects.

A Comprehensive Analysis of the Role of Oxidative Stress in the Pathogenesis and Chemoprevention of Oral Submucous Fibrosis.

Oral submucous fibrosis (OSMF) is a chronic oral potentially malignant disorder (OPMD). It is described as a scarring disease of the oral mucosa associated with excess oxidants and insufficient antioxidants. While it is becoming increasingly accepted that oxidative stress results in excessive accumulation of collagen and progressive fibrosis of the submucosal tissues, there is limited data regarding the moderation of oxidative stress to initiate or prevent OSMF. To assess the scope for mechanism-based approaches to prevent or reverse OSMF, we systematically evaluated the existing literature and investigated the role of oxidative stress in the pathogenesis and chemoprevention of OSMF. A search for relevant articles on PubMed and Scopus was undertaken using pre-defined inclusion and exclusion criteria. A total of 78 articles were selected in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) guidelines. The articles eligible for assessment investigated both OSMF and/or oxidative stress biomarkers or specific antioxidants. Both in vitro and human studies consistently demonstrated variations in oxidative stress biomarker levels in OSMF and revealed an increase in oxidative stress, paralleling the development of the disease. Furthermore, the use of antioxidant supplements was overall associated with an improvement in clinical outcomes. Having identified the significance of oxidative stress in OSMF and the therapeutic potential of antioxidant supplements, this scoping review highlights the need for further well-designed studies in the development of mechanism-based interventions for managing OSMF.

Predict the role of lncRNA in kidney aging based on RNA sequencing

BackgroundLong noncoding RNAs (lncRNAs) are involved in physiological and pathological processes. However, no studies have been conducted on the relationship between lncRNAs and renal aging.ResultsFirst, we evaluated the histopathology of young (3-month-old) and old (24-month-old) C57BL/6J mouse kidneys. Masson trichrome staining and PAS staining showed interstitial collagen deposition and fibrosis, mesangial matrix expansion, a thicker basement membrane and renal interstitial fibrosis in old mouse kidneys. Senescence-associated β-galactosidase (SA-β-gal)-positive areas in the kidneys of old mice were significantly elevated compared to those of young mice. Then, we analyzed the differential expression of lncRNAs and mRNAs in the kidneys of young and old mouse kidneys by RNA-seq analysis. 42 known and 179 novel differentially expressed lncRNAs and 702 differential mRNAs were detected in the mouse kidney. Next, we focused on the differentially expressed mRNAs and lncRNAs by RNA-seq. GO and KEGG analyses were performed based on differentially expressed mRNAs between young and old mouse kidneys. Transregulation based on RIsearch and the correlation coefficient of mRNA-lncRNA were also calculated. The mRNA-lncRNA network was constructed by choosing a Spearman correlation coefficient > 0.9 or <-0.9. GO and KEGG pathway enrichment analyses revealed that differentially expressed mRNAs participated in aging-related pathways. A total of 10 lncRNAs and trans-regulated mRNAs were constructed. Finally, we validated the role of lncRNA Gm43360 by CCK-8, flow cytometry, western blot and SA-β-gal staining. The expression level of Adra1a was positively correlated and Csnk1a1 was negatively correlated with lncRNA Gm43360. The cell counting kit-8 (CCK-8) results showed that lncRNA Gm43360 promoted cell viability. LncRNA Gm43360 increased the percentage of S phase cells and decreased the percentage of G1 phase cells compared with the negative control. LncRNA Gm43360 decreased the expression of p53, p21 and SA-β-gal.ConclusionsLncRNA Gm43360 may play a protective role in kidney aging.

Correlations Between the Expression of Stromal Cell Activation Related Biomarkers, L-NGFR, Phospho-ERK1-2 and CXCL12, and Primary Myelofibrosis Progression.

In myelofibrosis, pathologically enhanced extracellular matrix production due to aberrant cytokine signalling and clonal megakaryocyte functions result(s) in impaired hemopoiesis. Disease progression is still determined by detecting reticulin and collagen fibrosis with Gomori’s silver impregnation. Here, we tested whether the expression growth related biomarkers L-NGFR/CD271, phospho-ERK1-2 and CXCL12 can be linked to the functional activation of bone marrow stromal cells during primary myelofibrosis progression. Immunoscores for all tested biomarkers showed varying strength of positive statistical correlation with the silver impregnation based myelofibrosis grades. The intimate relationship between spindle shaped stromal cells positive for all three markers and aberrant megakaryocytes was likely to reflect their functional cooperation. L-NGFR reaction was restricted to bone marrow stromal cells and revealed the whole length of their processes. Also, L-NGFR positive cells showed the most intersections, the best statistical correlations with myelofibrosis grades and the strongest interrater agreements. CXCL12 reaction highlighted stromal cell bodies and a weak extracellular staining in line with its constitutive release. Phospho-ERK1-2 reaction showed a similar pattern to CXCL12 in stromal cells with an additional nuclear staining in agreement with its role as a transcription factor. Both p-ERK1-2 and CXCL12 were also expressed at a moderate level in sinus endothelial cells. Connexin 43 gap junction communication channels, known to be required for CXCL12 release to maintain stem cell niche, were also expressed progressively in the myelofibrotic stromal network as a support of compartmental functions. Our results suggest that, diverse growth related pathways are activated in the functionally coupled bone marrow stromal cells during myelofibrosis progression. L-NGFR expression can be a useful biological marker of stromal cell activation which deserves diagnostic consideration for complementing Gomori’s silver impregnation.

Relationship between first-line treatment response and bone marrow fibrosis in newly diagnosed multiple myeloma

Objectives: Multiple myeloma (MM) occurs with uncontrolled and clonal increase proliferation of plasma cells in the bone marrow. Myelofibrosis can be primary or can be secondary when associated with other malignant or non-malignant diseases. MM is a malignant disease in which both collagen and reticulin fibrosis can be detected together at the time of diagnosis.The aim of this study is to investigate the relationship between bone marrow fibrosis at diagnosis and response to after first-line treatment in newly diagnosed MM. Material and Methods: In this study, 95 newly diagnosed MM patients were analyzed retrospectively. Demographic characteristics, complete blood count, biochemical examinations, bone marrow fibrosis grades, and first-line treatment response of the patients were retrieved from records as it is a retrospective study. Patients were divided into 2 groups according to their response to first- line treatment. Patients who have a strict complete response (sCR), complete response (CR) or a very good partial response (VGPR) responses to first-line therapy were included in the first group. Patients who gave partial response (PR), minimal response (MR) or progressive disease (PD) responses to the first-line therapy were included in the second group. Results: There were 72 patients in the Group I (good response group) and 23 patients in Group II (poor response group). Between the good response group and poor response group myeloma type, platelet count at diagnosis, β2 microglobulin, lactate dehydrogenase, erythrocyte sedimentation rate, bone marrow plasma cell ratio, R-ISS, and first-line treatment were not statistically significant (P &gt; 0.05 ). Age was statistically significantly lower in the good response group (P = 0.04). In male gender, a better response was obtained (P = 0.02). At the time of diagnosis, hemoglobin levels in the good response group were found high compared to the poor response group (P = 0.02). Bone marrow fibrosis was found to be lower at the time of diagnosis in the group that responded good response to first-line treatment (P = 0.01). Conclusion: In this study, it was shown that bone marrow fibrosis at diagnosis is an important factor affecting the response to first-line treatment.The degree of bone marrow fibrosis detected at the time of diagnosis in MM may guide the selection of targeted therapy in first-line treatment.

Determination of cardiac fibrosis using picrosirius red fluorescence

Automatic measurement and quantification of collagen accumulation and fibrosis in the heart using multiphoton excitation methods is more difficult compared to other organs, as myosin and collagen both contribute to the second harmonic generation (SHG) signal, inhibiting any specificity of collagen only signals. Therefore, the most common fibrosis quantification using SHG often fails to quantify cardiac fibrosis. Picrosirius red (PSR) is a common dye that is used for the quantification of fibrosis using polarized light.

Gypenosides Attenuate Pulmonary Fibrosis by Inhibiting the AKT/mTOR/c-Myc Pathway.

Gypenosides (Gyps), the major active constituents isolated from Gynostemma pentaphyllum, possess anti-inflammatory and antioxidant activities. Previous studies have demonstrated that Gyps displayed potent ameliorative effects on liver fibrosis and renal fibrosis. In this study, we found that Gyps significantly reduced the mortality of bleomycin-induced pulmonary fibrosis mice (40% mortality rate of mice in the model group versus 0% in the treatment group). Masson staining showed that Gyps could reduce the content of collagen in the lung tissue of pulmonary fibrosis mice Masson staining and immunohistochemistry demonstrated that the expression of the collagen gene α-SMA and fibrosis gene Col1 markedly decreased after Gyps treatment. The active mitosis of fibroblasts is one of the key processes in the pathogenesis of fibrotic diseases. RNA-seq showed that Gyps significantly inhibited mitosis and induced the G2/M phase cell cycle arrest. The mTOR/c-Myc axis plays an important role in the pathological process of pulmonary fibrosis. RNA-seq also demonstrated that Gyps inhibited the mTOR and c-Myc signaling in pulmonary fibrosis mice, which was further validated by Western blot and immunohistochemistry. AKT functions as an upstream molecule that regulates mTOR. Our western blot data showed that Gyps could suppress the activation of AKT. In conclusion, Gyps exerted anti-pulmonary fibrosis activity by inhibiting the AKT/mTOR/c-Myc pathway.

Fibrosis and Collagen-I Accumulation in Bali Cattle Liver Tissue Infected with Fasciola gigantica

This study aims to determine the histopathological changes of fibrosis and accumulation of collagen I in liver tissue of Bali cattle infected with Fasciola gigantica in the Badung district, Bali Indonesia. On examination of 100 Bali cattle livers at the traditional abattoir of Badung, 37 of them were found to contain F. gigantica worms in both the bladder and bile ducts. The liver tissue was then fixed with 10% formalin for processing and stained with hematoxylin-eosin dyes. The liver sample preparation in the coating slide was also reacted with rabbit anti-collagen-I antibody. On histopathological examination, it was found that the distribution of fibrosis varied in the portal, interlobular, and bridging fibrosis areas. Immunohistochemical examination showed an immunoreactive reaction using rabbit anti-collagen I in the portal area with strong and weak intensity in the interlobular area and in bridging fibrosis. Fibrosis and accumulation of collagen I in Bali cattle infected with F. gigantica at the traditional abattoir of Badung district, Bali Indonesia was found in the portal, interlobular, and bridging fibrosis areas.

Human umbilical cord mesenchymal stem cells combined with pirfenidone upregulates the expression of RGS2 in the pulmonary fibrosis in mice

ObjectiveThe therapeutic effect of umbilical cord-derived mesenchymal stem cells (hUC-MSCs) in combination with pirfenidone (PFD) on pulmonary fibrosis in mice and its possible mechanism were investigated.MethodsC57BL/6 mice were randomly divided into six groups: control group, model group, P10 group, P30 group, P100 group, and P300 group. Modeled by tracheal intubation with 3 mg/kg bleomycin drip, each dose of PFD was administered daily by gavage from day 7 onwards. The mice were observed continuously for 21 days and survival was recorded. Lung tissues were collected on day 21, and hematoxylin–eosin (HE) and Masson staining were performed to assess morphological changes and collagen deposition in the lungs. Collagen content was measured by the Sircol method, and fibrosis marker levels were detected by PCR and Western blot. Another batch of C57BL/6 mice was then randomly divided into five groups: hUC-MSC control group, model group, P100 group, hUC-MSC treatment group, and hUC-MSCs + P30 group. On day 7, 5 × 105 hUC-MSCs were injected into the tail vein, the mice were administered PFD gavage daily from day 7 onwards, and their survival was recorded. Lung tissues were collected on day 21 to detect pathological changes, the collagen content, and the expression of regulator of G protein signaling 2 (RGS2). Pulmonary myofibroblasts (MFBs) were divided into an MFB group and an MFB + hUC-MSCs group; different doses of PFD were administered to each group, and the levels of RGS2, intracellular Ca2+, and fibrosis markers were recorded for each group.ResultsCompared with other PFD group doses, the P100 group had significantly improved mouse survival and lung pathology and significantly reduced collagen and fibrosis marker levels (p < 0.05). The hUC-MSCs + P30 group had significantly improved mouse survival and lung pathology, significantly reduced collagen content and fibrosis marker levels (p < 0.05), and the efficacy was better than that of the P100 and hUC-MSCs groups (p < 0.05). RGS2 expression was significantly higher in the MSCs + P30 group compared with the P100 and hUC-MSCs groups (p < 0.05). PFD increased RGS2 expression in MFBs (p < 0.05) in a dose-dependent manner. Compared with PFD and hUC-MSCs treatment alone, combination of hUC-MSCs and PFD increased RGS2 protein levels, significantly decreased intracellular Ca2+ concentration, and significantly reduced fibrosis markers.ConclusionThe findings suggest that hUC-MSCs combined with low-dose PFD have a therapeutic effect better than that of the two treatments used separately. Its effect on attenuating bleomycin-induced pulmonary fibrosis in mice is related to the increase of RGS2.

Semi-Digital Workflow of Removable Partial Denture Fabrication for Scleroderma-Induced Microstomia Patients: Two Clinical Reports

Scleroderma is a chronic connective tissue disease characterized by collagenous fibrosis resulting in the hardening and contracture of the skin and mucosa. Most of the scleroderma patients develop Raynaud&#39;s phenomenon that the fingers and toes feel numb, prickly and frigid in response to cold temperatures or stress. Due to fibrosis of the skin and soft tissues, scleroderma induces microstomia that clinically represents limited mouth opening which results in difficulty both for the patients a

Comparative pathomorphological characteristics of idiopathic pulmonary fibrosis and fibrotic hypersensitivity pneumonitis

The paper presents an X-ray morphological differential diagnosis of idiopathic pulmonary fibrosis (IPF) and fibrotic hypersensitivity pneumonitis (FHP). It describes the etiology, pathogenesis, radiological signs, and pathoanatomy of IPF and FHP. For differential diagnosis, radiological and morphological signs were studied in 105 patients with IPF and in 111 patients with FHP. The mean ages of patients with IPF or FHP were 65.0±8.9 and 48.9±12.3 years, respectively. The history of IPF to the moment of its diagnosis ranged from 1 to 18 months, while that of FHP was 35 to 79 days. The authors describe the additional morphological signs of FHP: delicate collagen fibrosis; smooth muscle metaplasia in the interalveolar septa and fibrotic areas; fibroblastic foci mainly in the walls of bronchioles; plasma cell infiltration of interalveolar septa with a touch of neutrophils and eosinophils. A table has been compiled for differential diagnosis according to the morphological signs of IPF and FHP.

The Place of Histochemical Stains for Differential Diagnosis in Duodenal Biopsies

The biopsy is the primary examination method in diagnosing duodenal pathologies and Gluten enteropathy that can cause symptoms such as gas, diarrhea, weight loss, anemia, osteoporosis. Allows monitoring of response to therapy. Histochemical studies are inevitable in specific findings such as reactive changes, giardia, subepithelial collagen thickening, fibrosis, and gastric metaplasia. We aimed to understand the contribution of histochemical applications to differential diagnosis and to show their superiority compared to hematoxylin-eosin sections in routine use by simultaneously making histomorphological evaluations on hematoxylin and eosin sections. This research is a retrospective study conducted in 2011, using the archives of Yüzüncü Yıl University, Faculty of Medicine, Department of Pathology, between 2001-2010. Ten normal, 50 Gluten enteropathy and 50 duodenitis samples; Along with histopathological examinations, we applied Masson Trichrome, Periodic acid-Schiff, Periodic acid-Schiff with diastasis, Periodic acid Schiff-Alcian Blue, High Iron Diamine-Alcian Blue. Chi-square, likelihood ratio tests, and SPSS (ver: 13) statistical package program used. We detected microscopic findings parallel to diagnostic criteria. While the number of goblet cells and intraepithelial neutrophils did not differ in the groups (P =0.176 and P=0.096), there was a significant variation in the intraepithelial lymphocyte count (P=0.010). The frequency of flattening and blunting of the villi, crypt hyperplasia, and Brunner gland hyperplasia were significant in gluten enteropathies (P=0.000). Conclusion We could not find any difference in specific pathologies associated with histochemical studies in our group, so hematoxylin-eosin sections are more valuable in the differential diagnosis.

Reduced M2 macrophages and adventitia collagen dampen the structural integrity of blood blister-like aneurysms and induce preoperative rerupture.

ObjectiveBlood blister–like aneurysms (BBAs) are extremely rare aneurysms. They are predisposed to preoperative rerupture with a high case‐fatality rate. Here, we attempt to interrogate the distinct clinicopathology and the histological basis underlying its clinical rerupture.MethodsThree middle meningeal arteries, 11 BBA (5 reruptured, 6 non‐rerupture) and 19 saccular aneurysm samples were obtained for histopathological investigation. Three reruptured BBAs, 3 non‐reruptured BBAs and 6 saccular (3 ruptured, 3 unruptured) aneurysms were obtained for quantitative flow cytometry analysis.ResultsCompared with true saccular aneurysms, the BBA aneurysm wall lacks arterial stroma cells including CD31+ endothelial cells and α‐SMA + smooth muscle cells. Only fibroblasts and adventitial collagen were observed in the BBA aneurysm wall. Meanwhile, BBAs were enriched with infiltrated inflammatory cells, especially polarized macrophages. Based on the rerupture status, those reruptured BBAs showed drastically reduced fibroblasts and adventitia collagen. Moreover, M2‐polarized macrophages were observed dominant in BBAs and exhibit repairing cellular functions based on their interplays with arterial fibroblasts. Reduced M2 macrophages and arterial tissue repairing modulation may be responsible for the decreasing collagen synthesis and fibrosis repairment, which potentially dampens the aneurysm integrity and induces BBA aneurysm reruputre.ConclusionsBBAs poses histopathological features of occult pseudoaneurysms or dissecting aneurysms. Reduced M2 macrophages and adventitia collagen may dampen the structural integrity of BBAs and induce preoperative rerupture.

Histopathological Evaluation of Burn Injury

Background: Burns always have posed a threat to the sensitive human body. It is a common catastrophe today as burn injury cases are one of the common emergencies admitted to any hospital. In a medico-legal setting, the main difficulty arises when a burnt body found in unknown circumstances is brought for post-mortem examination. In such a case, a forensic expert has to opine whether the burn injuries sustained are antemortem or post-mortem. It is also of paramount importance to determine the age of burn injuries sustained. Aging of burn injuries by the naked eye is sometimes difficult and unreliable. By histopathological examination of burnt tissue, one can predict whether the victim was alive or dead when the fire was going on and determine the age of burn injuries. Materials and Methods: The present cross-sectional study was carried out in the Department of Forensic Medicine and Toxicology, Pt. B.D.Sharma PGIMS, Rohtak. Total 3033 autopsies were conducted in the mortuary of the Department of Forensic Medicine andToxicology, PGIMS, Rohtak, from November 5, 2011`to November 5, 2013. During that period, out of 486 cases reported burns, randomly 100 cases were selected for this study. Results and Conclusions: In the present study, efforts were made to determine burn wound vitality and age by histopathological evaluation. The study revealed that the inflammatory infiltrate consisted mainly of polymorphonuclear leucocytes in initial stages (up to 3rd-day post-burn) with gradual lymphocytes, macrophages, and fibroblasts forming immature granulation tissue. On the 14th-day post-burn, the burn wound revealed repair by regeneration, increased collagen deposition, fibrosis, and granulation tissue formation. It was infiltrated with fibroblasts, macrophages, and lymphocytes. Our study demonstrated that evaluation of various histopathological changes occurring during the distinctive phases of burn wound healing help in the estimation of its vitality and age.

Recovery from Liver Failure and Fibrosis in a Rat Portacaval Anastomosis Model after Neurointermediate Pituitary Lobectomy.

Liver diseases, including cirrhosis, viral hepatitis, and hepatocellular carcinoma, account for approximately two million annual deaths worldwide. They place a huge burden on the global healthcare systems, compelling researchers to find effective treatment for liver fibrosis-cirrhosis. Portacaval anastomosis (PCA) is a model of liver damage and fibrosis. Arginine vasopressin (AVP) has been implicated as a proinflammatory-profibrotic hormone. In rats, neurointermediate pituitary lobectomy (NIL) induces a permanent drop (80%) in AVP serum levels. We hypothesized that AVP deficiency (NIL-induced) may decrease liver damage and fibrosis in a rat PCA model. Male Wistar rats were divided into intact control (IC), NIL, PCA, and PCA+NIL groups. Liver function tests, liver gene relative expressions (IL-1, IL-10, TGF-β, COLL-I, MMP-9, and MMP-13), and histopathological assessments were performed. In comparison with those in the IC and PCA groups, bilirubin, protein serum, and liver glycogen levels were restored in the PCA+NIL group. NIL in the PCA animals also decreased the gene expression levels of IL-1 and COLL-I, while increasing those of IL-10, TGF-β, and MMP-13. Histopathology of this group also showed significantly decreased signs of liver damage with lower extent of collagen deposition and fibrosis. Low AVP serum levels were not enough to fully activate the AVP receptors resulting in the decreased activation of cell signaling pathways associated with proinflammatory-profibrotic responses, while activating cell molecular signaling pathways associated with an anti-inflammatory-fibrotic state. Thus, partial reversion of liver damage and fibrosis was observed. The study supports the crucial role of AVP in the inflammatory-fibrotic processes and maintenance of immune competence. The success of the AVP deficiency strategy suggests that blocking AVP receptors may be therapeutically useful to treat inflammatory-fibrotic liver diseases.