Elastica Van Gieson Staining Research Articles

LncRNA SENCR suppresses abdominal aortic aneurysm formation by inhibiting smooth muscle cells apoptosis and extracellular matrix degradation.

Abdominal aortic aneurysm (AAA) is a progressive chronic dilatation of the abdominal aorta without effective medical treatment. This study aims to clarify the potential of long non-coding RNA SENCR as a treatment target in AAA. Angiotensin II (Ang-II) was used to establish AAA model in vitro and in vivo. Reverse transcription quantitative PCR and western blot were performed to measure the expression of SENCR and proteins, respectively. Annexin V-FITC/PI double staining was carried out to detect the apoptotic rate in vascular smooth muscle cells (VSMCs), and cell apoptosis in aortic tissues was determined by TUNEL staining. Besides, hematoxylin and eosin and Elastica van Gieson staining were performed for histological analysis of aortic tissues. SENCR was downregulated in AAA tissues and Ang-II-stimulated VSMCs. Overexpression of SENCR could inhibit Ang-II-induced VSMC apoptosis, while inhibition of SENCR facilitated Ang-II-induced VSMC apoptosis. Moreover, the expression of matrix metalloproteinase (MMP)-2 and MMP-9 in Ang-II-induced VSMCs was reduced following SENCR overexpression, while tissue inhibitor of metalloproteinases 1 (TIMP-1) expression was increased. In vivo, overexpression of SENCR improved the pathological change in aortic tissues and the damage in arterial wall elastic fibers induced by Ang-II, as well as suppressed Ang-II-induced cell apoptosis and extracellular matrix degradation. Overall, SENCR was decreased in AAA. Overexpression of SENCR inhibited AAA formation via inhibition of VSMC apoptosis and extracellular matrix degradation. We provided a reliable evidence for SENCR acting as a potential target for AAA treatment.

Defining the Histologic Support Structures of the Nasal Ala and Soft Triangle: Toward Understanding the Cause of Iatrogenic Alar Retraction.

As rhinoplasty techniques have evolved to more extensive dissections, the incidence of iatrogenic deformities, such as alar rim retraction, has risen. Its mechanism is presently unknown. This study examined the microscopic anatomy of the nasal ala to define architectural support elements at the histologic level to determine why rhinoplasty dissection creates such deformities. Eight cadaveric noses were harvested and sectioned through the soft triangle and ala. Various tissue stains were performed. Slides were examined using light microscopy. Anatomical features pertaining to cartilage, skin, mucosa, elastic fibers, and muscle were documented. Four male and four female noses were sectioned. The median cadaver age was 64 years (range, 47 to 83 years). On Elastica van Gieson stain, distinct elastic fibers span from the vestibular lining to the caudal margin of the lower lateral cartilage, and from the caudal edge of the lower lateral cartilage to the external alar skin. In the nasal ala midsection, trichrome stains reveal that skeletal muscle is located far beyond the lower lateral cartilage, close to the free alar margin. The soft triangle shows a distinct microanatomical structure, with heavy longitudinal condensations of elastin. These histologic findings have not been previously reported. A distinct anatomical alar wall endoskeleton has been identified. It is obligatorily disrupted by specific rhinoplasty maneuvers when dissection is carried out over the lateral crura and into areas without cartilaginous support. This microanatomy may explain factors that contribute to postoperative alar wall retraction. Leaving this area undisturbed or performing adjunctive measures with rhinoplasty can provide structural support to the external valves, thus minimizing the risk of deformity.

Wall shear stress analysis using 17.6 Tesla MRI: A longitudinal study in ApoE-/- mice with histological analysis.

This longitudinal study was performed to evaluate the feasibility of detecting the interaction between wall shear stress (WSS) and plaque development. 20 ApoE-/- mice were separated in 12 mice with Western Diet and 8 mice with Chow Diet. Magnetic resonance (MR) scans at 17.6 Tesla and histological analysis were performed after one week, eight and twelve weeks. All in vivo MR measurements were acquired using a flow sensitive phase contrast method for determining vectorial flow. Histological sections were stained with Hematoxylin and Eosin, Elastica van Gieson and CD68 staining. Data analysis was performed using Ensight and a Matlab-based “Flow Tool”. The body weight of ApoE-/- mice increased significantly over 12 weeks. WSS values increased in the Western Diet group over the time period; in contrast, in the Chow Diet group the values decreased from the first to the second measurement point. Western Diet mice showed small plaque formations with elastin fragmentations after 8 weeks and big plaque formations after 12 weeks; Chow Diet mice showed a few elastin fragmentations after 8 weeks and small plaque formations after 12 weeks. Favored by high-fat diet, plaque formation results in higher values of WSS. With wall shear stress being a known predictor for atherosclerotic plaque development, ultra highfield MRI can serve as a tool for studying the causes and beginnings of atherosclerosis.

Left hemidiaphragmatic elevation: Value of histology

Abstract Two neonates presented with left hemidiaphragmatic elevation. Case 1 was a boy transferred to our center at age 3 d. Asymptomatic congenital diaphragmatic eventration (CDE) was suspected. At 8 months, chest computed tomography (CT) revealed persistent left hemidiaphragmatic elevation, and surgical treatment was performed. Immunohistochemistry (IHC) of a resected membranous part of the left hemidiaphragm showed that it contained desmin-positive muscle fibers. Collagen bundles were detected by Elastica van Gieson (EvG) staining. Case 2 was a boy delivered at 38 weeks 3 d gestational age. Fetal ultrasonography and MRI had suggested left hemidiaphragmatic elevation, suspicious for CDE. His Apgar scores were 8 and 6 at 1 minute and 5 minutes. He was intubated immediately after delivery. Weaning from ventilation was unsuccessful and surgery was performed 21 d after delivery. Partial resection of the membranous part and repair of the muscular hemidiaphragm were performed. EcG staining revealed collagen bundles, even though desmin-positive muscle fibers were not seen on IHC. Muscle fibers and collagen bundles are two important components of the diaphragm, and are not found in either the pleura or peritoneum. Accordingly, CDE was diagnosed in both cases. EvG suggested the diagnosis.

Grating-based phase-contrast CT (PCCT): histopathological correlation of human liver cirrhosis and hepatocellular carcinoma specimen

AimsTo correlate signal intensities in grating-based phase-contrast CT (PCCT) images obtained at a synchrotron light source and a conventional X-ray source with tissue components in human liver cirrhosis and hepatocellular...

Histological investigation of picosecond laser-toning and fractional laser therapy

Rejuvenation therapy using picosecond pulse laser and picosecond pulsed fractional therapy with a fractional lens have been performed with clinical effects evaluated. However, no histological analysis of effects on photoaged skin exists. In this study, influence of laser-toning and fractional therapy using picosecond pulse laser on photoaging was histologically investigated. The flexor side forearm of a male, age 61, with photoaging was divided into three 20 cm2 areas and irradiated with approximately 400 shots of 10-Hz laser, 8 mm spot size, and nine passes at an output of 0.7, 0.9, and 1.1 J/cm2 using picosecond laser-toning therapy six times, every two weeks. Two weeks post final irradiation, 2 mm punch biopsies were taken from the irradiation fields. Fractional therapy using Micro Lens Array (MLA) attached picosecond fractional therapy was applied to the medial crural skin with marked photoaging of a male, age 63. Irradiation was applied at 0.5 and 0.7 J/cm2 through two passes, with 3 mm punch biopsies taken from each irradiation field immediately after and again two months post-irradiation. Samples were subjected to hematoxylin and eosin (HE) and Elastica van Gieson staining and compared. In the picosecond laser-toning therapy sample, photoaging-induced dermis reconstruction occurred. The picosecond fractional therapy sample showed both epidermis and dermis reconstruction, with intrinsic aging and photoaging improvements. Recovery of dermal and epidermal age related atrophy by picosecond laser-toning and picosecond fractional therapy was histologically confirmed. Picosecond fractional therapy demonstrated superior improvement.

Lipid Core Plaque Distribution Using Near-infrared Spectroscopy Is Consistent with Pathological Evaluation in Carotid Artery Plaques.

Carotid artery stenting (CAS) is performed as a treatment for carotid artery stenosis. However, lipid-rich plaques cause embolic complications and sequelae. Near-infrared spectroscopy (NIRS) can identify lipid components by applying a near-infrared absorption pattern, and the distribution of lipid components can be evaluated as the maximum lipid core burden index (maxLCBI). Intravascular ultrasound (IVUS) equipped with NIRS has been clinically applied recently, and its diagnostic usefulness and validation have been reported for coronary arteries; however, its consistency with actual pathological diagnosis in carotid artery lesions has not been validated. In this study, we investigated the consistency between the maxLCBI values and histopathological diagnoses. Patients with cervical carotid artery stenosis who underwent carotid endarterectomy (CEA) were examined in this prospective study. Pathological diagnosis was determined after NIRS evaluation, which was performed on the extracted plaques ex vivo. The histological slices of decalcified and paraffin-embedded sections were stained by hematoxylin–eosin (HE) and Elastica van Gieson (EVG), and for low-density lipoprotein (LDL), C-reactive protein (CRP), CD68, and glycophorin A. The correlation between maxLCBI values and histological findings. Seventy lesions assessed by NIRS were pathologically analyzed. There was a positive linear correlation between maxLCBI values and pathological findings as determined by HE (angle), HE (area%), EVG, CRP, and CD68 staining (respectively, r = 0.624, p <0.001; r = 0.578, p <0.001; r = 0.534, p <0.001; r = 0.723, p <0.001; r = 0.653, p <0.001). In conclusion, the maxLCBI values assessed by NIRS showed a significant positive linear correlation with pathological evaluations in carotid lesions. The maxLCBI values in carotid arteries are consistent with pathological evaluations.

Influence of caveolin-1 and endothelial nitric oxide synthase on adventitial inflammation in aortic transplants

Restenosis after endovascular interventions is a clinically relevant process that is directly associated with increased morbidity. Thereby, an increased migration and proliferation of vascular smooth muscle cells (VSMCs) is mainly responsible for recurrent lumen narrowing. Previously, we showed that caveolin‑1 (Cav‑1) and endothelial nitric oxide synthase (eNOS) were directly involved in neointimal proliferation. In the current study, we investigated the impact of Cav‑1 and eNOS on adventitial processes in a murine model. Denuded aortas from C57Bl6n (wild‑type [WT]), Cav‑1-/, eNOS-/, and Cav‑1-//eNOS-/ mice were transplanted into common carotid arteries of WT mice. The explantation was performed after 6 weeks, followed by Elastica van Gieson staining and immunohistochemistry. The Cav‑1-/ and the eNOS-/ aortas showed an increase in the adventitial content of macrophages, whereas their combined knockout did not lead to additive effects. Differences were observed despite the same acceptor, suggesting the local origin of inflammatory cells. Furthermore, the WT transplants exhibited the highest content of vascular endothelial growth factor A (VEGF‑A) despite the lowest macrophage content. In contrast, the knockout aortas showed a decreased content of VEGF‑A as well as decreased expression of α-smooth muscle actin (α-‑SMA) in the tunica media, suggesting induced VSMC migration. Moreover, the WT aortas exhibited increased neovessel formation. Cav‑1 and eNOS inhibit adventitial macrophage‑derived inflammation and modulate its cellular function. The knockout of Cav‑1 and eNOS leads to a decreased expression of VEGF-A, with decreased neovessel formation and increased migration of VSMCs, which promote a proatherogenic phenotype.

Cerebral cavernomas in adults and children express relaxin.

To shed light on the role of relaxin in cerebral cavernous malformations (CCMs) in adults and children, the authors investigated endothelial cell (EC) expression of relaxin 1, 2, and 3; vascular endothelial growth factor receptor-1 and -2 (VEGFR-1 and -2); Ki-67; vascular geometry; and hemorrhage, as well as the clinical presentation of 32 patients with surgically resected lesions. Paraffin-embedded sections of 32 CCMs and 5 normal nonvascular lesion control (NVLC) brain tissue samples were immunohistochemically stained with antibodies to relaxin 1, 2, and 3; angiogenesis growth factor receptors Flt-1 (VEGFR-1) and Flk-1 (VEGFR-2); and proliferation marker Ki-67. For morphometric analysis, Elastica van Gieson stain was used, and for hemorrhage demonstration, Turnbull stain was used. Data from the pediatric and adult CCMs were compared with each other and with those obtained from the NVLCs. Statistical analyses were performed with Fisher's exact test, the chi-square test, the phi correlation coefficient, and the Student t-test. A p value < 0.05 was considered significant. Pediatric and adult cavernoma vessels did not significantly differ in diameter. Hemorrhage was observed in CCMs but not in NVLC samples (p < 0.05). There was no difference in expression of Ki-67, VEGFR-1 and -2, and relaxin 1, 2, and 3 in the ECs of pediatric and adult CCMs. The ECs of CCMs were largely negative for relaxin 3 compared to NVLCs (p < 0.05), whereas CCMs, compared to control brain tissue samples, more frequently expressed Flt-1 and relaxin 2 (p < 0.05). Ki-67 was not expressed in the NVLCs, but the difference was not statistically significant. Relaxin 1 and 2 expression and increased expression of VEGFR-1 were associated with a supra- versus infratentorial location (p < 0.05). Relaxin 1 and 2 and VEGFR-1 play a role in supratentorial cavernomas. Relaxin 3 may play a physiological role in normal brain vasculature. Relaxin 1 and 3 are also found in normal cerebral vasculature. Relaxin 1, 2, and 3 are associated with increased VEGFR-1 expression.

MS17.05 Controversies in Pathologic Staging

MS17.05 Controversies in Pathologic Staging

An intra-amniotic injection of mesenchymal stem cells promotes lung maturity in a rat congenital diaphragmatic hernia model.

We aimed to evaluate the effect of human mesenchymal stem cells (hMSCs) on congenital diaphragmatic hernia (CDH) by intra-amniotic injection in a rat CDH model. Nitrofen (100mg) was administered to pregnant rats at E9.5. hMSCs (1.0 × 106) or PBS was injected into each amniotic cavity at E18, and fetuses were harvested at E21. The fetal lungs were classified into normal, CDH, and CDH-hMSCs groups. To determine the lung maturity, we assessed the alveolar histological structure by H&E and Weigert staining and the alveolar arteries by Elastica Van Gieson (EVG) staining. TTF-1, a marker of type II alveolar epithelial cells, was also evaluated by immunohistochemical staining and real-time reverse transcription polymerase chain reaction. The survival rate after intra-amniotic injection was 72.1%. The CDH-hMSCs group had significantly more alveoli and secondary septa than the CDH group (p < 0.05). The CDH-hMSCs group had larger air spaces and thinner alveolar walls than the CDH group (p < 0.05). The medial and adventitial thickness of the pulmonary artery in the CDH-hMSCs group were significantly better (p < 0.001), and there were significantly fewer TTF-1-positive cells than in the CDH group (p < 0.001). These results suggest that intra-amniotic injection of hMSCs has therapeutic potential for CDH.

Abstract 794: Endothelial Specific Ablation of Estrogen Receptor Alpha Rapid Signaling Revealed Exacerbated Vascular Remodeling Response

Background: Estrogen exerts complex physiological effects via its rapid (non-genomic) and genomic actions. In particular, rapid signaling of estrogen receptor alpha (ERα) has been implicated in the vasculo-protective effects, in which both endothelial and smooth muscle cells might be involved. However, no prior studies have determined the role of ERα rapid signaling in the endothelium. This study aims to clarify the impact of ERα rapid signaling in the vasculo-protection, using a novel mouse model lacking endotheilal-specific ERα rapid signaling. Methods and Results: In immunoblotting, p85α and pGSK3β, non-genomic pathway downstream signals, were reduced in ERα mutants RR259/260AA with estradiol (E2) stimulation at concentrations of 10 -11 and 10 -10 M . ERE-luciferase assay demonstrated E2 induced genomic pathway activity was preserved even in ERα mutants RR259/260AA. We generated a mouse model in which rapid signaling of ERα was ablated in the endothelium by crossing Tie2-Cre transgenic mice with floxed ERα mutants (RR 259/260 AA) in which p85α and ERα interaction was disrupted. In endothelial cells isolated from ERα KI/KI Tie2 cre/+ animals, E2 at a concentration of 10 -8 M failed to induce phosphorylation of Akt, confirming the absence of ERα rapid signaling. Baseline characteristics at 8 to 12 weeks of age were undistinguishable between the genotypes, including body weight (KI:19.2±0.366 g vs WT: 18.9±0.356 g ) systolic blood pressure (KI:106.1±3.336 vs WT: 103.5±2.133 mmHg) and echocardiographic fractional shortening (KI:54.4±0.751 vs WT: 55.19±0.947 %). We then assessed how vascular remodeling process was impacted in a carotid artery wire injury model. Histological analyses with Elastica van Gieson staining two weeks after induction of injury revealed that wall thickness and area of medial layer composed mainly of smooth muscle cells were significantly increased in ERα KI/KI Tie2 cre/+ mouse, as compared to wild types (KI:44.85±1.948 μm vs WT: 29.32±1.753 μm , KI:42,047±1,560 μm 2 vs WT: 32,815±1,211 μm 2 , respectively ). Conclusions: Our results demonstrate that the rapid signaling of ERα in the endothelium critically regulates vascular smooth muscle cell growth after vascular injury, suggesting the essential role to vascular remodeling.

Giant cell arteritis without cranial manifestations caused mesenteric involvement: a case report

BackgroundGiant cell arteritis (GCA) is a granulomatous vasculitis and targets large vessels with predominance for the aortic arch and the cranial branches. GCA with cranial symptoms shows headache, jaw claudication, and ophthalmologic symptoms and thus was previously called temporal arteritis. Recently, cases of GCA without cranial manifestations and extracranial GCA have been reported.Case presentationA 76-year-old woman was referred to our hospital complaining of sudden abdominal pain and high fever. Her present history of illness did not show any cranial symptoms such as headache, visual disturbance, or stroke. CT images showed severe thickening of the small intestinal mesentery and massive ascites. She was diagnosed to have acute abdomen probably with gastrointestinal perforation and underwent the emergent laparotomy. Excisions of a 60-cm length of the jejunum including the thickening mesenteric lesion were carried out. Marked hypertrophy of the vascular intima and mild stenosis of the arterial lumen were displayed with infiltration of lymphocytes, neutrophils, and eosinophils. Scattered multinucleated giant cells on the endothelium, in the intima, media, and adventitia were demonstrated. Elastica van Gieson stain showed focal loss and fragmentation of the internal elastic lamina. Histopathological examinations showed typical GCA. Her postoperative process was uneventful without any symptoms, and she was followed as an out-patient prescribed with daily doses of 40 mg of prednisolone.ConclusionsWe hereby report a rare case of mesenteric involvement in GCA without cranial manifestations and elucidate the histopathological features of extracranial GCA in arteries as well as veins and jejunum.

Pathological evaluation of the visceral pleura in the radical pleurectomy/decortication for malignant pleural mesothelioma patients.

Radical pleurectomy/decortication (P/D) is applied as a surgical treatment of resectable malignant pleural mesothelioma (MPM). Although P/D removed visceral pleura, dissection plain was not histologically explored previously. We examined a pathological evaluation of surgically removed visceral pleura in P/D. Twenty-five patients with MPM who underwent P/D at the Tokyo Medical and Dental University Hospital between April 2010 and April 2018 were studied. The 25 cases included 20 with epithelioid tumors, 4 with a biphasic tumor and 1 with desmoplastic tumors. Nine, 1, 11 and 4 patients had mesotheliomas of stages I, II, III and IV, respectively. We analyzed the site of the visceral pleural lesions using the HE & Elastica van Gieson (EVG) staining. The tumor involvement of the pleura and the surgical dissection plane were defined using the depth criteria, D0-3. We added survival analyses according to the depth criteria. Ninety-nine lesions in total 45 lobes: 20 upper right, 14 middle, 20 lower, 22 upper left, and 23 lower were examined. Based on the depth D 0-3 criteria, there were 21 type D0, 18 type D1, 22 type D2 and 38 type D3 lesions. The growth of tumor cells in the pleura was partially diffuse or nodular in all cases. While 38 lesions which invaded the lung parenchyma were excised, another 61 lesions that reached within the pleura were dissected from lung parenchyma. Type D2&3 showed poor survivals than type D0&1. The lung parenchyma was always the dissection plane in P/D, regardless of tumor involvement in the visceral pleura. The depth criteria would help us in classifying pleural invasion histologically and possibly predicting the prognosis.

SR1001 confers protection against the experimental abdominal aortic aneurysm induced by angiotensin II in ApoE-/-mice

Objective To investigate the effect and mechanism of SR1001 treatment in angiotensin Ⅱ (AngⅡ) induced experimental abdominal aortic aneurysm (EAAA) in apolipoprotein E (ApoE-/-) mice. Methods Forty-five male apolipoprotein E-deficient (ApoE-/-) mice were randomly divided into the following 3 groups: Sham group, AngⅡ model group (Control group), AngⅡ+ SR1001 treatment group (SR1001 group). All mice were treatment with 0.5% dimethylsurfoxide (DMSO) or SR1001 by intraperitoneal injection. The aortic diameter was measured via ultrasound imaging, and the suprarenal aorta was chosen as the measuring point on days 0, 14, 28 thereafter. Twenty-eight days after AngⅡ infusion, the abdominal aortic specimens were harvested. The elastica van Gieson (EVG) staining, and immunofluorescent staining were performed for histopathologic analysis. The protein levels of interleukin (IL)-17, monocyte chemotactic protein (MCP)-1, interferon (IFN)-γ were determined by Western blotting analysis. Results (1) Compared with the Control group, the AAA incidence in the SR1001 group was significantly reduced (33.3% vs. 73.3%, χ2=4.821, P=0.028), Abdominal aorta diameter [(1.39±0.06) mm vs. (1.98±0.11) mm, P=0.009]. However, there was no significant difference in abdominal aorta diameter between the SR1001 group and Sham group [(1.39±0.06) mm vs. (1.07±0.08) mm, P=0.064]. (2) SR1001 treatment in SR1001 group preserves the structure of the aortic wall (P=0.018). (3) SR1001 treatment in SR1001 group attenuates local inflammatory cell infiltration in the EAAA model. (4) SR1001 treatment in SR1001 group suppresses the protein expression of pro-inflammatory mediators (IL-17, MCP-1, IFN-γ) in the EAAA model (P=0.008, 0.016, 0.023, respectively). Conclusion Treatment with SR1001 inhibits the Th17/IL-17A-related inflammatory responses, preserves the structure of the aortic wall, reduces local inflammatory cell infiltration and inflammatory factors expression, and attenuates the progression of abdominal aortic aneurysm. Key words: SR1001; Angiotensin Ⅱ; Apolipoprotein E-/-mice; Interleukin-17; Inflammatory cell; Experimental abdominal aortic aneurysm

Xanthine Oxidase Inhibition by Febuxostat in Macrophages Suppresses Angiotensin II-Induced Aortic Fibrosis.

BACKGROUNDSeveral reports from basic researches and clinical studies have suggested that xanthine oxidase (XO) inhibitors have suppressive effects on cardiovascular diseases. However, the roles of a XO inhibitor, febuxostat (FEB), in the pathogenesis of vascular remodeling and hypertension independent of the serum uric acid level remain unclear.METHODSTo induce vascular remodeling in mice, angiotensin II (Ang II) was infused for 2 weeks with a subcutaneously implanted osmotic minipump. FEB was administered every day during Ang II infusion. Aortic fibrosis was assessed by elastica van Gieson staining. Mouse macrophage RAW264.7 cells (RAW) and mouse embryonic fibroblasts were used for in vitro studies.RESULTSFEB suppressed Ang II-induced blood pressure elevation and aortic fibrosis. Immunostaining showed that Ang II-induced macrophage infiltration in the aorta tended to be suppressed by FEB, and XO was mainly colocalized in macrophages, not in fibroblasts. Transforming growth factor-β1 (TGF-β1) mRNA expression was induced in the aorta in the Ang II alone group, but not in the Ang II + FEB group. Ang II induced α-smooth muscle actin-positive fibroblasts in the aortic wall, but FEB suppressed them. XO expression and activity were induced by Ang II stimulation alone but not by Ang II + FEB in RAW. FEB suppressed Ang II-induced TGF-β1 mRNA expression in RAW.CONCLUSIONSOur results suggested that FEB ameliorates Ang II-induced aortic fibrosis via suppressing macrophage-derived TGF-β1 expression.

Histological study of the thin membranous structure made of dense connective tissue around the esophagus in the upper mediastinum

The structure of the fascia in upper mediastinum has already been reported from gross anatomical viewpoints by Sarrazin. But it is necessary to understand meticulous anatomy for thoracoscopic or mediastinoscopic surgery. So herein, we investigate histologically the thin membranous structure made of dense connective tissues. Semi-sequential transverse sections of the mediastinum were obtained from three cadavers. Hematoxylin and eosin staining, Elastica van Gieson staining, and Masson trichrome staining were performed to identify the presence and location of the thin membranous structure made of dense connective tissues. The "visceral sheath" and "vascular sheath," as previously described by Sarrazin, were observed histologically. These two thin membranous structures do not surround the esophagus and trachea cylindrically. In addition, the "visceral sheath" on the right side of the upper mediastinum was unclear in comparison to the left side. The "visceral sheath" (on the left side) gradually became unclear, and seemed to almost disappear; the esophagus was found to be very close to the thoracic duct on the caudal side of the bifurcation of the trachea. Although the left recurrent nerve was located inside the "visceral sheath" in all cadavers, the left recurrent nerve lymph nodes were located inside the "visceral sheath" in cadaver 1 and between the "visceral sheath" and "vascular sheath" in cadaver 3. The "visceral sheath" around the esophagus in the upper mediastinum was histologically demonstrated; however, the findings were not constant.

Fractional CO2 laser treatment for vaginal laxity: A preclinical study.

Various studies have investigated treatment for vaginal laxity with microablative fractional carbon dioxide CO2 laser in humans; however, this treatment has not yet been studied in an animal model. Herein, we evaluate the therapeutic effects of fractional CO2 laser for tissue remodeling of vaginal mucosa using a porcine model, with the aim of improving vaginal laxity. The fractional CO2 laser enables minimally invasive and non-incisional procedures. By precisely controlling the laser energy pulses, energy is sent to the vaginal canal and the introitus area to induce thermal denaturation and contraction of collagen. We examined the effects of fractional CO2 laser on a porcine model via clinical observation and ultrasound measurement. Also, thermal lesions were histologically examined via hematoxylin-eosin staining, Masson's trichrome staining, and Elastica van Gieson staining and immunohistochemistry. The three treatment groups, which were determined according to the amount of laser-energy applied (60, 90, and 120 mJ), showed slight thermal denaturation in the vaginal mucosa, but no abnormal reactions, such as excessive hemorrhaging, vesicles, or erythema, were observed. Histologically, we also confirmed that the denatured lamina propria induced by fractional CO2 laser was dose-dependently increased after laser treatment. The treatment groups also showed an increase in collagen and elastic fibers due to neocollagenesis and angiogenesis, and the vaginal walls became firmer and tighter because of increased capillary and vessel formation. Also, use of the fractional CO2 laser increased HSP (heat shock protein) 70 and collagen type I synthesis. Our results show that microablative fractional CO2 laser can produce remodeling of the vaginal connective tissue without causing damage to surrounding tissue, and the process of mucosa remodeling while under wound dressings enables collagen to increase and the vaginal wall to become thick and tightened. Lasers Surg. Med. 50:940-947, 2018. © 2018 Wiley Periodicals, Inc.

Prevalence and Prognostic Significance of Extramural Venous Invasion in Patients with Locally Advanced Esophageal Cancer

BackgroundExtramural venous invasion (EMVI) is a known adverse prognostic factor in patients with colorectal carcinoma. The prevalence and significance of EMVI in esophageal cancer (EC) patients is still unclear.MethodsFrom a prospectively maintained database, we retrospectively reviewed the resection specimens of patients with pathologic locally advanced (pT3/T4/N0-3) EC who were treated with curative intent between 2000 and 2015. Patients with previous malignancies and gastroesophageal junction (type II/III) tumors were excluded. Included were 81 patients who underwent surgery alone and 37 patients who underwent neoadjuvant chemoradiotherapy (nCRT). EMVI was assessed on hematoxylin and eosin slides and confirmed or excluded by additional Elastica van Gieson staining. Survival was analyzed using a multivariable Cox regression.ResultsEMVI was present in 23.5% (n = 19) of patients in the surgery-alone group and 21.6% (n = 8) of patients in the nCRT group. The prevalence of EMVI after surgery alone was significantly high in squamous cell carcinomas and among tumors located in the mid-esophagus, as well as those with lymphovascular invasion (p < 0.05). After nCRT, the presence of EMVI was significantly high in tumors with lymphovascular and perineural tumor growth (p = 0.034). EMVI status was an independent adverse prognostic factor for disease-free survival [hazard ratio (HR) 7.0, 95% confidence interval (CI) 2.3–21.8; p =0.001] and overall survival (HR 6.5, 95% CI 2.2–19.1; p = 0.001) in the surgery-alone group for node-positive tumors.ConclusionsIn this study of locally advanced > pT3/N0-3 EC patients, EMVI was present in 23.5% of patients in the surgery-alone group and in 21.6% of patients after nCRT. EMVI was an independent adverse prognostic factor in patients after surgery alone.

Vibratory function and healing outcomes after small intestinal submucosa biomaterial implantation for chronic vocal fold scar.

Vocal fold scar is a major cause of dysphonia, and optimal treatments do not currently exist. Small intestinal submucosa (SIS) is a biomaterial developed for the treatment of a variety of pathologies. The purpose of this study was to investigate the effects of SIS implantation on tissue remodeling in scarred vocal folds using routine staining, immunohistochemistry, and high-speed videoendoscopy (HSV). Prospective, blinded group analysis. Thirteen New Zealand White rabbits underwent a vocal fold scarring procedure followed by microflap elevation with or without SIS implantation. Seven months later, they underwent a phonation procedure with HSV and laryngeal harvest. Alcian blue and elastica van Gieson staining and immunohistochemistry for collagen types I and III were used to evaluate histological healing outcomes. Dynamic functional remodeling of the scarred vocal fold in the presence of SIS implants was evaluated using HSV imaging to capture restoration of vibratory amplitude, amplitude ratio, and left-right phase symmetry. Density of collagen I was significantly decreased in SIS versus microflap-treated vocal folds. No differences were found between groups for hyaluronic acid, elastin, or collagen type III. Organization of elastin in the subepithelial region appeared to affect amplitude of vibration and the shape of the vocal fold edge. SIS implantation into chronic scar reduced the density of collagen I deposits. There was no evidence of a negative impact or complication from SIS implantation. Regardless of treatment type, organization of elastin in the subepithelial region may be important to vibratory outcomes. NA. Laryngoscope, 128:901-908, 2018.