Elastic Stain Research Articles

Application of TTF1 immunohistochemistry combined with elastic fiber double staining in the diagnosis of lung adenocarcinoma

Application of TTF1 immunohistochemistry combined with elastic fiber double staining in the diagnosis of lung adenocarcinoma

The application of cytokeratin 7 and elastic fiber double staining with automation platform in the diagnosis of lung adenocarcinoma

为更好地显示肺腺癌中弹力纤维的变化及其与肿瘤细胞的关系，应用全自动免疫组织化学和全自动特殊染色平台对30例肺腺癌组织细胞进行角蛋白（CK）7和弹力纤维双重染色，结果显示在双染切片中CK7免疫组织化学的特异度和灵敏度与单独染色基本没有差异，两者相结合除可以特异性显示胸膜受侵的情况，还观察到多数厚壁血管弹力纤维的增生和血管壁的重构，以及更多CK7和弹力纤维单独染色难以观察到的肿瘤细胞早期浸润血管壁的现象。.

Changes in the expression levels of elastic fibres in yak lungs at different growth stages

BackgroundYaks have a strong adaptability to the plateau environment, which can be attributed to the effective oxygen utilization rate of their lung tissue. Elastic fibre confers an important adaptive structure to the alveolar tissues in yaks. However, little research has been focused on the structural development of lung tissues and the expression levels of elastic fibres in yaks after birth. Therefore, this study aimed to investigate the morphological changes of elastic fibers and expression profiles of fibre-formation genes in yak lungs at different growth stages and the relationship between these changes and plateau adaptation.ResultsHistological staining was employed to observe the morphological changes in the lung tissue structure of yaks at four different ages: 1 day old, 30 days old, 180 days old and adult. There was no significant difference in the area of a single alveolus between the 1-day-old and 30-day-old groups (P-value > 0.05). However, the single alveolar area was gradually increased with an increase in age (P-value < 0.05). Elastic fibre staining revealed that the amount of elastic fibres in alveolar tissue was increased significantly from the ages of 30 days to 180 days (P-value < 0.05) and stabilized during the adult stage. Transcriptome analysis indicated that the highest levels of differentially expressed genes were found between 30 days of age and 180 days of age. KEGG analysis showed that PI3K-Akt signalling pathway and MAPK pathway, which are involved in fibre formation, accounted for the largest proportion of differentially expressed genes between 30 days of age and 180 days of age. The expression levels of 36 genes related to elastic fibre formation and collagen fibre formation were also analysed, and most of these genes were highly expressed in 30-day-old and 180-day-old yaks.ConclusionsThe content of elastic fibres in the alveolar tissue of yaks increases significantly after birth, but this change occurs only from 30 days of age to 180 days of age. Our study indicates that elastic fibres can improve the efficiency of oxygen utilization in yaks under harsh environmental conditions.

Hidradenitis suppurativa lesions are associated with decreased collagen, decreased elastin, and increased neovascularization

ABSTRACT Hidradenitis suppurativa (HS) is a dermatological condition characterized by ruptured hair follicles inducing a localized, but massive chronic inflammatory response. Analysis of 104 HS lesions in formalin-fixed paraffin-embedded tissues identified the most common histologic features found associated with HS were migrating epithelial sheets, acute and chronic inflammation, perivascular chronic inflammation in regions adjacent to active lesions, and granulation tissue. Additionally, using a modified Masson’s trichrome stain principally for collagen, Verhoeff van Gieson elastic stain, and CD34 immunohistochemistry assay for endothelium, lesional areas in the dermis with chronic inflammation exhibited a significant decrease in collagen and elastin, and an increase in neovascularization.

Modification of Pathologic T Classification for Non-small Cell Lung Cancer With Visceral Pleural Invasion: Data From 1,055 Cases of Cancers ≤ 3 cm

Visceral pleural invasion (VPI) with PL1 or PL2 increases the T classification from T1 to T2 in non-small cell lung cancers (NSCLCs)≤ 3cm. We proposed a modified T classification based on VPI to guide adjuvant therapy. Is it reasonable to upstage PL1-positive cases from T1 to T2 for NSCLCs≤ 3cm? In total, 1,055 patients with resected NSCLC were retrospectively included. Tumor sections were restained with hematoxylin and eosin stain and Victoria blue elastic stain for the elastic layer. Disease-free survival (DFS) and overall survival (OS) were calculated by the Kaplan-Meier method. Subgroup analysis and a Cox proportional hazards model were used to further determine the impact of VPI on survival. The extent of VPI was diagnosed as PL0 in 824 patients, PL1 in 133 patients, and PL2 in 98 patients. The 5-year DFS rates of patients with PL0, PL1, and PL2 were 62.6%, 60.2%, and 28.8%(P< .01), whereas the corresponding 5-year OS rates were 78.6%, 74.4%, and 50.0%(P< .01), respectively. As predicted, the DFS and OS of patients with PL2 were much worse than those of patients with PL0 (P< .01) and PL1 (P< .01). However, both the DFS and OS of patients with PL0 and PL1 were comparable (DFS: P= .198; OS: P= .150). For node-negative cases, the DFS and OS of patients with PL0 and PL1 were also comparable (DFS: P= .468; OS: P= .388), but patients with PL2 had much worse DFS and OS than patients with PL0 (P< .01) and PL1 (P< .01). Multivariable analyses suggested that PL2, together with node positivity and poor cell differentiation, was an independent adverse prognostic factor. In NSCLCs≤ 3cm, tumors with PL1 should remain defined as T1, not T2. Overtreatment by adjuvant chemotherapy in node-negative NSCLCs≤ 3cm might be avoided in PL1 cases.

Effects of Ambroxol Hydrochloride on Lung Tissue Cell Apoptosis and Vascular Remodeling in Rats with Smoke Induced Chronic Obstructive Pulmonary Disease

To evaluate the effects of ambroxol hydrochloride on lung tissue cell apoptosis and vascular remodeling in rats with smoke induced chronic obstructive pulmonary disease. Sprague Dawley rats were randomly divided into normal, model, experimental and control groups (n=20). The rat model of chronic obstructive pulmonary disease was established by fumigation using Yan’an Cigarettes for 64 d. After fumigation, the experimental and control groups were subcutaneously injected with 5 mL of Ambroxol hydrochloride (20 mg/kg) and 5 mL of Bambuterol hydrochloride (20 mg/kg) respectively, while the normal and model groups were intraperitoneally injected with an equal dose of normal saline. Following drug intervention for 28 d, the pathological changes in lung tissues, vascular remodeling, lung tissue cell apoptosis, expressions of alpha-smooth muscle actin and vascular endothelial growth factor, as well as levels of caspase-3, B-cell lymphoma-2 and B-cell lymphoma-2 associated X protein were detected by hematoxylin and eosin staining, elastic Van Gieson staining, terminal deoxynucleotidyl transferase dUTP nick end labeling staining, immunohistochemical staining and western blotting, respectively. Compared with normal group, the damage of lung tissues was obvious, the medial thickness of pulmonary arterioles significantly increased, the degree of vascular muscularization, apoptosis rate and expressions of alpha-smooth muscle actin, vascular endothelial growth factor, caspase-3 and B-cell lymphoma-2 associated X protein rose and the expression of B-cell lymphoma-2 decreased in model group (p<0.05). Compared with model group, the damage of lung tissues was significantly improved, the medial thickness of pulmonary arterioles decreased, the degree of vascular muscularization, apoptosis rate and expressions of alpha-smooth muscle actin, vascular endothelial growth factor, caspase-3 and B-cell lymphoma-2 associated X protein reduced and the expression of B-cell lymphoma-2 rose in experimental and control groups (p<0.05). Ambroxol hydrochloride can inhibit the apoptosis of lung tissue cells and improve vascular remodeling, thereby protecting the lung tissues of chronic obstructive pulmonary disease rats.

Chronic Intermittent Hypoxia Regulates CaMKII-Dependent MAPK Signaling to Promote the Initiation of Abdominal Aortic Aneurysm.

Obstructive sleep apnea (OSA) is highly prevalent in patients with abdominal aortic aneurysm (AAA). However, the effects of OSA on AAA initiation in a murine model of sleep apnea have not been completely studied. In this paper, Apoe−/− C57BL/6 mice infused with angiotensin II (Ang II) were placed in chronic intermittent hypoxia (CIH) condition for inducing OSA-related AAA. CIH significantly promoted the incidence of AAA and inhibited the survival of mice. By performing ultrasonography and elastic Van Gieson staining, CIH was found to be effective in promoting aortic dilation and elastin degradation. Immunohistochemical and zymography results show that CIH upregulated the expression and activity of MMP2 and MMP9 and upregulated MCP1 expression while downregulating α-SMA expression. Also, CIH exposure promoted ROS generation, apoptosis, and mitochondria damage in vascular smooth muscle cells (VSMCs), which were measured by ROS assay, TUNEL staining, and transmission electron microscopy. The result of RNA sequencing of mouse aortas displayed that 232 mRNAs were differently expressed between Ang II and Ang II+CIH groups, and CaMKII-dependent p38/Jnk was confirmed as one downstream signaling of CIH. CaMKII-IN-1, an inhibitor of CaMKII, eliminated the effects of CIH on the loss of primary VSMCs. To conclude, a mouse model of OSA-related AAA, which contains the phenotypes of both AAA and OSA, was established in this study. We suggested CIH as a risk factor of AAA initiation through CaMKII-dependent MAPK signaling.

The estimation of age from elastic fibers in the tunica media of the aortic wall in a thai population: a preliminary study using aorta image analysis.

Image analysis has an increasing role in the identification of individuals in forensic application. Beside the bones, microstructural of arteries can be used in age estimation study. Aorta is the largest elastic artery which consists of many elastic fibers. Elastin in arterial wall highly resist to chemical and physical influence. The purposes of the study were to quantify elastic fibers in tunica media in each location of the aorta and examine the correlation between elastic fibers and age by using image analysis program. A total of 36 human aortas were dissected in 4 locations. The aortas were obtained from cadavers with an age range of 20 to 90 years. Specimens were stained with Elastic Van Gieson staining. Histological images were investigated about elastic fibers using light microscope with cellSens program and aorta image analysis was used for the evaluation of data. The results showed that the mean percentage density of elastic fibers in the ascending aorta and the aortic arch increased. However, the mean percentage density of elastic fibers decreased in the 31 to 40 years age group in the thoracic aorta and the abdominal aorta and decreased in each location of aorta continuously until 81 to 90 years. The abdominal aorta showed the highest correlation with age (r=0.732) followed by the thoracic aorta, the aortic arch and the ascending aorta, respectively. Changes in the percentage density of elastic fibers in the tunica media of the aortic wall can be used to add information to age estimation for identification purposes.

Variability in Synoptic Reporting of Colorectal Cancer pT4a Category and Lymphovascular Invasion.

Serosal involvement (pT4a category) and lymphovascular invasion have prognostic significance in colorectal carcinoma, but are subject to interobserver variation in assessment. To provide the first large-scale assessment of interobserver variability in pT4a category and lymphovascular invasion reporting in real-world practice and to explore the impact of information from guidelines on variability in reporting these features. Analysis of 1555 consecutive synoptic reports of colorectal carcinoma was performed using multivariate logistic regression. Interobserver variability before and after the presentation of guideline information was assessed using an image-based survey. Significant differences in the odds of reporting pT4a versus pT3 category, detecting lymphovascular invasion of any type, and detecting large vessel invasion were identified among hospital sites and for individual pathologists compared with the median pathologist at the same site. Consistent with these results, interobserver agreement was only moderate in the image-based survey regarding T4a staging and lymphovascular invasion (all κ ≤ 0.57). The provision of information from guidelines did not tend to increase interobserver agreement in the survey, though responses in favor of using an elastic stain increased following recommendations for their use. However, when observers were provided with elastic-stained images, interobserver agreement remained only moderate (κ = 0.55). Real-world reporting of pT4a category and lymphovascular invasion shows substantial variability at both local and regional levels. Our study underscores the need to address these features in quality initiatives, and provides a novel method through which existing synoptic data can be harnessed to monitor reporting patterns and provide individualized feedback.

Histological and ultrastructural changes of the colon in dextran sodium sulfate-induced mouse colitis.

Ulcerative colitis (UC) is a complex disease that results from a dysregulated immune response in the gastrointestinal tract. A mouse model orally administered with dextran sodium sulfate (DSS) is the most widely used experimental animal model of UC. However, the ultrastructure of the colon in mouse colitis is poorly understood. In the present study, colonic specimens from DSS-induced UC mice underwent hematoxylin and eosin staining, Masson's trichrome staining and Verhoeff's elastic staining. In addition, the ultrastructure of samples was examined by transmission electron microscopy. UC was successfully induced by 7 consecutive days of DSS oral administration. The goblet cell architecture of the UC tissue was damaged in the mucosa. Additionally, a significant number of inflammatory cells infiltrated into the stroma and the structure of the intestinal gland was destroyed. The tissue in the submucosa showed significant edema. Hyperplasia was also identified in the submucosa, promoting a disorganized microstructure within the colon wall. Numerous collagen fibers in the muscular layer were disrupted, and the fiber bundles were thinner compared with those in the normal control group. Furthermore, in the DSS-induced UC group, the smooth muscle cell showed edema, the cell membrane structure was unclear and the shape of the nucleus was irregular. In conclusion, the present study revealed important histological and ultrastructural changes in the colon of DSS-induced UC mice. These features may contribute to improved understanding of the pathogenesis and mechanism of UC.

Efficacy and limitations of the histological diagnosis of type 1 autoimmune pancreatitis with endoscopic ultrasound-guided fine needle biopsy with large tissue amounts

ObjectivesWe examined the efficacy and limitations of acquiring large specimens by endoscopic ultrasound-guided fine needle biopsy (EUS-FNB) for diagnosing type 1 autoimmune pancreatitis (AIP). MethodsPatients from 12 institutions with non-neoplastic diseases or pancreatic ductal adenocarcinoma (PDAC) with large EUS-FNB specimens were investigated. Slides stained with hematoxylin-eosin, elastic, IgG4, and IgG stains were evaluated. The IgG4- and IgG-positive cell numbers were counted in three foci. The diagnoses were based on the Japan Pancreas Society 2011 (JPS 2011) criteria and the International Consensus Diagnostic Criteria (ICDC). ResultsWe analyzed 85 non-neoplastic (definite type 1 AIP in 73/85 based on the ICDC) cases and 64 PDAC cases. IgG4-positive cells were numerous (>10 in 85.9%), and the IgG4/IgG ratios were high (>40% in 81.2%). Plasma cell crushing by an artifact caused unsuccessful immunostaining, notably in smaller samples. Tissue lengths were an important factor for the presence of storiform fibrosis and obliterative phlebitis, but storiform fibrosis was equivocal even in large tissues. A definite or possible histological diagnosis was achieved in 45.9% (39/85) and 41.2% (35/85), respectively, and contributed to the definite final diagnosis of type 1 AIP in 33.3% (ICDC) and 55.6% (JPS 2011) in cases with segmental/focal lesions. In the PDAC group, >10 IgG4-positive cells was rare (2/58), but elastic stains revealed fibrous venous occlusions in 10.3% (6/58). ConclusionsEUS-FNB with large tissue amounts was useful for diagnosing type 1 AIP, notably by facilitating successful IgG4 immunostaining, but definite diagnosis may not be achieved even in cases with large specimens.

An analytical study of neocartilage from microtia and otoplasty surgical remnants: A possible application for BMP7 in microtia development and regeneration.

To investigate auricular reconstruction by tissue engineering means, this study compared cartilage regenerated from human chondrocytes obtained from either microtia or normal (conchal) tissues discarded from otoplasties. Isolated cells were expanded in vitro, seeded onto nanopolyglycolic acid (nanoPGA) sheets with or without addition of bone morphogenetic protein-7 (BMP7), and implanted in nude mice for 10 weeks. On specimen harvest, cartilage development was assessed by gross morphology, histology, and RT-qPCR and microarray analyses. Neocartilages from normal and microtia surgical tissues were found equivalent in their dimensions, qualitative degree of proteoglycan and elastic fiber staining, and quantitative gene expression levels of types II and III collagen, elastin, and SOX5. Microarray analysis, applied for the first time for normal and microtia neocartilage comparison, yielded no genes that were statistically significantly different in expression between these two sample groups. These results support use of microtia tissue as a cell source for normal auricular reconstruction. Comparison of normal and microtia cells, each seeded on nanoPGA and supplemented with BMP7 in a slow-release hydrogel, showed statistically significant differences in certain genes identified by microarray analysis. Such differences were also noted in several analyses comparing counterpart seeded cells without BMP7. Summary data suggest a possible application for BMP7 in microtia cartilage regeneration and encourage further studies to elucidate whether such genotypic differences translate to phenotypic characteristics of the human microtic ear. The present work advances understanding relevant to the potential clinical use of microtia surgical remnants as a suitable cell source for tissue engineering of the pinna.

Elastin and collagen IV double staining: A refined method to detect blood vessel invasion in breast cancer.

Blood vessel invasion (BVI) is a prognostic indicator in various cancers. Elastic stain, which highlights blood vessel walls, is commonly used to detect BVI. In the breast, however, its diagnostic usefulness is limited because it also highlights some intraductal carcinoma components, which often mimic BVI. In this study, we aimed to improve BVI detection in breast cancer and developed a double staining: Victoria blue for elastin and immunohistochemistry for collagen IV. Collagen IV fibers were retained along the basement membranes of intraductal carcinoma components, whereas they were rearranged or lost in BVI. From these observations, we defined BVI as the presence of tumor cells inside an elastic ring with a rearrangement or loss of collagen IV fibers. Using these criteria, we found BVI in 148 cases (49%) among 304 cases of primary operable invasive breast carcinoma, and the presence of BVI correlated significantly with poor prognosis. By contrast, we detected BVI in 94 cases (31%) or 14 cases (5%) by elastic van Gieson or CD31 immunostaining among the same cases, respectively, with no statistically significant association with prognosis. Thus, elastin and collagen IV double staining facilitates the detection of BVI in breast cancer and is useful to predict prognosis.

Identification of a Primary Stroma and Novel Endothelial Cell Projections in the Developing Human Cornea.

PurposeTo investigate the initial events in the development of the human cornea, focusing on cell migration, and extracellular matrix synthesis and organization. To determine whether elastic fibers are present in the extracellular matrix during early human corneal development.MethodsHuman corneas were collected from week 7 to week 17 of development. An elastic fiber-enhancing stain, tannic acid–uranyl acetate, was applied to all tissue. Three-dimensional serial block-face scanning electron microscopy combined with conventional transmission electron microscopy was used to analyze the corneal stroma.ResultsAn acellular collagenous primary stroma with an orthogonal arrangement of fibrils was identified in the central cornea from week 7 of corneal development. At week 7.5, mesenchymal cells migrated toward the central cornea and associated with the acellular collagenous matrix. Novel cell extensions from the endothelium were identified. Elastic fibers were found concentrated in the posterior peripheral corneal stroma from week 12 of corneal development.ConclusionsThis study provides novel evidence of an acellular primary stroma in the early development of the embryonic human cornea. Cell extensions exist as part of a communication system and are hypothesized to assist in the migration of the mesenchymal cells and the development of the mature cornea. Elastic fibers identified in early corneal development may play an important role in establishing corneal shape.

Modified Intravascular Interventional Operation of Balloon-Bound Elastase Induction Method to Establish Rabbit Aneurysm Model Using Cardiovascular Angiography

Objective: In this paper, we establish a simple, efficient and stable rabbit aneurysm model using modified intravascular interventional operation of balloon-bound elastase induction method. Methods: We selected 20 healthy adult New Zealand white rabbits. An aortic aneurysm model of rabbit right common carotid artery was established by modified intravascular interventional balloon-binding elastase induction method using cardiovascular angiography. The cerebral angiography confirmed the model successfully after 4 weeks. Some animals were given paraffin-embedded H-E staining and elastic fiber staining after sacrifice. Angiography was performed for all experiments to build our models and to confirm our findings. Results: Two rabbits died during postoperative feeding, and the remaining 18 rabbits were cultured for 4 weeks and confirmed by angiography to make an aneurysm model. The success rate of aneurysm was 90%, the diameter of the tumor-bearing artery was 3.12±0.20, the short diameter of the tumor was 3.41±0.88, the long diameter of the tumor was 6.30±1.65, and the width of the neck was 3.49±0.80. HE pathology showed that the destruction of the elastic layer of the aneurysm disappeared, similar to the pathological changes of human aneurysms. Conclusions: The rabbit model of aneurysm was established by modified intravascular intervention combined with elastase induction based on angiography. It is similar to human aneurysm in morphology, histology and hemodynamics. The method is simple, the damage is small, the aneurysmal formation rate is high, and the performance is high. Stable and reliable, low production cost, it is an ideal animal model for the development of preclinical neurological interventional devices.

Eruptive Cutaneous Collagenoma: Report of Two Cases

Abstract Introduction: Eruptive cutaneous collagenoma is non familial connective tissue nevi of unknown etiology presented with an abrupt onset. To date, the literatures on eruptive cutaneous collagenoma are extremely rare in china. Here, we report two women with eruptive cutaneous collagenoma. Case present: Two women presented with multiple asymptomatic, skin-colored papules and nodules on the trunk and extremities with no systemic involvement. Histopathology revealed dense, coarse collagen fibers by hematoxylin-esoin stain and decreased, fragmented elastic fibers by Elastic stain compared with the normal skin. Basing on these findings, the diagnosis of eruptive cutaneous collagenoma was made. No specific treatment was given. Discussion: Eruptive cutaneous collagenoma is a very rare dermatosis that is often misdiagnosed as other connective tissue nevi. The pathogenesis is unclear, and also no efficient treatment is available. It is usually diagnosed based on clinical and histopathological findings. Conclusion: The present cases are relatively rare type of eruptive cutaneous collagenoma that provide more experience for clinician, and may be helpful for them to make correct diagnosis for suspicious cases.

Early Intervention of Tongxinluo () on Right Ventricular Function Assessed by Echocardiography in Rats with Pulmonary Arterial Hypertension Induced by Monocrotaline.

To investigate the effect of early intervention of Tongxinluo (, TXL) on right ventricular function (RVF) of rats with pulmonary arterial hypertension (PAH) induced by monocrotaline (MCT). A total of 30 adult male Sprague-Dawley rats were assigned to 5 groups with complete random experiment design: Sham group (Sham), MCT group, TXL group, sildenafil (SIL) group and combination group (TXL+SIL), 6 rats in each group. Rats were injected with 50 mg/kg MCT solution for inducing PAH model except for those in the sham group. From the day of modeling, rats of TXL, SIL and TXL+SIL groups were given TXL (1.2 g/kg), SIL (10 mg/kg) and combination solution (TXL:1.2 g/kg, SIL: 10 mg/kg) respectively, and rats in Sham and MCT groups were given normal saline (5 mL/kg). The samples were collected and tested after 21 consecutive days of intragastric administration. Echocardiography was used to measure the related indices of RVF, including pulmonary arterial flow spectrum, pulmonary artery diameter (PAD), right ventricular wall thickness (RVWT), right ventricular diameter (RVD), tricuspidannular plane systolic excursion (TAPSE), right atrium transverse diameter (RAT), and inferior vena cava diameter (IVCD). Elastic Verhoeff-Van Gieson staining was adopted to measure the percentage of wall thickness (WT%) of pulmonary arteriols. Hematoxylin-eosin staining was used to measure the cross-sectional area (CSA) of right ventricular cardiomyocytes. MCT-induced PAH rat model was successfully established. In MCT group the wall of pulmonary arterioles exhibited a prominent-increase thickness, PAD, RVWT, RVD, RAT, IVCD, WT%, right ventricular hypertrophy index (RVHI) as well as CSA of RV cardiomyocyte significantly increased (all P<0.01), and TAPSE markedly decreased (P<0.01). At the same time, TXL prominently improved all of the above indices (all P<0.01). In comparison with SIL, TXL significantly reduced RVD (P<0.05) and decreased CAS of RV cardiomyocytes (P<0.01), but TAPSE in SIL group was much larger than in TXL group (P<0.01). Moreover, TAPSE in TXL+SIL group was larger than that in TXL group (P<0.01), while the two groups performed equally well in terms of the other indices. Early intervention of TXL could inhibit pulmonary arterioles remodeling, and improve RVF by attenuating right ventricular hypertrophy, and TXL has a stronger effect on inhibiting right ventricular remodeling than SIL.

The sympathetic transmitter norepinephrine inhibits VSMC proliferation induced by TGFβ by suppressing theexpression of the TGFβ receptor ALK5 in aorta remodeling.

The sympathetic system is involved in the arterial diseases, but its mechanism remains poorly understood. The present study aimed to explore the impact of the sympathetic neurotransmitter norepinephrine (NE) on transforming growth factor (TGF) β signaling and the role of NE in aortic remodeling. Guanethidine was used to induce a regional chemical sympathetic denervation (CSD) in angiotensin II (AngII) and β-aminopropionitrile (BAPN)-induced aortic aneurysm models. The diameter of the aorta was measured, and elastic fiber staining was performed. TGFβ type I receptor kinase (ALK5) expression in rat aortic NE-treated vascular smooth muscle cells (VSMCs) was detected by reverse transcription-quantitative PCR and western blotting. The effects of NE and ALK5 overexpression on migration, proliferation, apoptosis and TGFβ signaling were also evaluated. Furthermore, adrenergic receptor blockers were used to determine which receptor was involved in the modulation on TGFβ signaling by NE. The results of the present study demonstrated that CSD protected rats from AngII+BAPN-induced aortic remodeling and aneurysm formation. Compared with the control group, NE inhibited VSMC proliferation and migration, but promoted apoptosis by suppressing ALK5 expression, reversing the effects of TGFβ signaling through the suppression of the SMAD-dependent canonical pathway and promotion of the non-canonical pathway. These effects were prevented by ALK5 overexpression. The inhibition of α- or β-adrenergic receptors alleviated the NE-mediated suppression of ALK5 expression. In conclusion, regional CSD protected rats from aortic aneurysm. NE inhibited SMAD2/3-dependent TGFβ signaling by suppressing ALK5 expression, which may serve an important role in VSMC biological functions. Both α- and β-adrenergic receptors were involved in the regulation of ALK5 expression by NE. Abnormal sympathetic innervation of the aorta may be used as a therapeutic target in aortic diseases.

Effect of Long-term Administration of Prostaglandin E1 on Morphologic Changes in Ductus Arteriosus

To improve survival of patients with hypoplastic left heart syndrome, combination therapy with bilateral pulmonary artery banding and prostaglandin E1 (PGE1)-mediated ductal patency was developed as an alternative for high-risk neonates in Japan. However, the effect of long-term PGE1 administration on ductus arteriosus remains unclear. Synchrotron radiation-based X-ray phase-contrast tomography (XPCT) enables clear visualization of soft tissues at an approximate spatial resolution of 12.5 μm. We aimed to investigate morphologic changes in ductus arteriosus after long-term PGE1 infusion using XPCT. Seventeen ductus arteriosus tissue samples from patients with hypoplastic left heart syndrome were obtained during the Norwood procedure. The median duration of lipo-prostaglandin E1 (lipo-PGE1) administration was 48 days (range, 3 to 123). Structural analysis of ductus arteriosus was performed and compared with conventional histologic analysis. The XPCT was successfully applied to quantitative measurements of ductal media. Significant correlation was found between the duration of lipo-PGE1 infusion and mass density of ductal media (R= 0.723, P= .001). The duration of lipo-PGE1 administration was positively correlated with elastic fiber staining (R= 0.799, P < .001) and negatively correlated with smooth muscle formation (R=-0.83, P < .001). No significant increase in intimal cushion formation was found after long-term lipo-PGE1 administration. Expression of ductus arteriosus dominant PGE2-receptor EP4 almost disappeared in specimens when lipo-PGE1 was administered over 3 days. Disorganized elastogenesis and little intimal cushion formation after long-term lipo-PGE1 administration suggest that ductus arteriosus remodeled to the elastic artery phenotype. Because EP4 was downregulated and ductus arteriosus exhibited elastic characteristics, the dosage of lipo-PGE1 might be decreased after a definite administration period.

The effect of the peritoneal tumor microenvironment on invasion of peritoneal metastases of high-grade serous ovarian cancer and the impact of NEOADJUVANT chemotherapy.

Peritoneal metastases of high-grade serous ovarian cancer (HGSOC) are small-sized deposits with superficial growth toward the peritoneal cavity. It is unknown whether integrity of the peritoneal elastic lamina (PEL) correlates with the peritoneal tumor microenvironment (pTME) and whether neoadjuvant chemotherapy (NACT) affects the pTME. We explored integrity of PEL, composition of pTME, effects of NACT, and the prognostic implications in patients with extensive peritoneal metastases of HGSOC. Peritoneal samples (n = 69) were collected during cytoreductive surgery between 2003 and 2016. Clinical data were collected from medical charts. Integrity of PEL was evaluated with elastic stains. T cell (CD3, CD8) and M2-macrophage markers (CD163) were scored using algorithms created in definiens tissue studio. Patients with a disrupted PEL (n = 39; 57%), more often had residual disease after surgery (p = 0.050), compared to intact PEL. An intact PEL was associated with increased intraepithelial (ie) CD8+ cells (p = 0.032), but was not correlated with improved survival. After NACT, increased ieCD3+ cells were shown, compared to no-NACT (p = 0.044). Abundance of total CD3+ and CD8+ cells were associated with PFS (multivariate HR 0.40; 95%CI 0.23-0.69 and HR 0.49; 95%CI 0.29-0.83) and OS (HR 0.33; 95%CI 0.18-0.62 and HR 0.36; 95%CI 0.20-0.64). M2-macrophage infiltration was not correlated with survival. NACT increases abundance of ieCD3+ cells in peritoneal metastases of HGSOC. Increase of CD3+ and CD8+ cells is associated with improved PFS and OS. This suggests that CD3+ and CD8+ cells may function as prognostic biomarkers. Their role as predictive biomarker for chemotherapy or immunotherapy response in HGSOC warrants further research.