Perichondrial progenitor cells promote proliferation and chondrogenesis of mature chondrocytes

Autologous chondrocytes are effective sources of cell therapy for engineering cartilage tissue to repair chondral defects, such as degenerative arthritis. The expansion of cells with chondrocyte characteristics has become a major challenge due to inadequate donor sites and poor proliferation of mature chondrocytes. The perichondrial progenitor cells (P cells) from the cambium layer of the perichondrium possessed significantly higher mesenchymal stem cell markers than chondrocytes (C cells). In the transwell co-culture system, P cells increased the passaging capacity of C cells from P6 to P9, and the cell number increased 128 times. This system increased the percentage of Alcian blue-positive chondrocytes from 40% in P6 to 62% in P9, contributing about 198 times more Alcian blue-positive chondrocytes than the control group. C cells co-cultured with P cells also exhibited higher proliferation than C cells cultured with P cell-conditioned medium. Similar results were obtained in nude mice that were subcutaneously implanted with C cells, P cells, or a mixture of the two cell types, in which the presence of both cells enhanced neocartilage formation in vivo. In aggregate, P cells enhanced the proliferation of C cells in a dose-dependent manner and prolonged the longevity of mature chondrocytes for clinical applications.

Teat sinus and duct adenomatous hyperplasia in dogs

Seventeen lesions diagnosed as teat sinus and duct adenomatous hyperplasia were identified in 10 dogs. All of the dogs were small breeds. Six were spayed female and 4 were male, 3 castrated and 1 intact. In 5 cases, the lesions involved multiple teats. They were pink to black, flattened to round, and sometimes crusted. Histologically, the lesions were usually pigmented (16/17), plaque-like to nodular masses composed of polygonal cells arranged in anastomosing trabeculae and bilayered ducts and/or cysts, with a fibrous to mucinous (Alcian blue-positive) stroma and squamous cysts (12/17). Scattered epithelial cells contained single, discrete, clear cytoplasmic vacuoles. Atypia was mild, and the mitotic count per 2.37 mm2 varied from 0 to 15 (average 2.7). Immunohistochemistry was performed on 14 of the lesions from 8 dogs. Epithelial cells were 100% panCK+ and included basally located CK14+/CK5_6+/p63+/calponin− cells and nonbasal CK19+/CK7+ cells. Cells manifesting squamous differentiation were usually panCK+/CK14+/CK5_6+/CK19−/CK7−/p63±/calponin−. In addition to fibroblasts, vimentin positivity was found in disseminated, round to stellate stromal and intraepithelial cells that often had black, granular, cytoplasmic pigment (consistent with dendritic/phagocytic cells and/or melanocytes). Of the 8 dogs for which clinical follow-up information was available, all were still alive and well, with no significant teat changes, development of mammary lesions or other masses 4 to 22 months (median 12.5) after biopsy. The histologic, immunohistochemical, and clinical findings were consistent with teat duct and sinus adenomatous hyperplasia. This is an uncommon, benign proliferative lesion that can involve multiple teats of female and male, small breed dogs.

LncRNA NONHSAT030515 promotes the chondrogenic differentiation of human adipose-derived stem cells via regulating the miR-490-5p/BMPR2 axis

Background Chondrogenic differentiation of human adipose-derived stem cells (hADSCs) is important for cartilage generation and degradation. LncRNAs play an essential role in stem cell differentiation. However, the role and mechanism of lncRNA in hADSCs remain unclear. Our previous study showed that miR-490-5p was downregulated during chondrogenic differentiation of hADSCs. In this study, we investigated the effect and mechanism of lncRNA NONHSAT030515 interacting with miR-490-5p on chondrogenic differentiation of hADSCs. Methods Alcian blue staining was used to assess the deposition of chondromatrix proteins following chondrogenic differentiation of human adipose stem cells. Immunohistochemistry was used to evaluate the expression of collagenII. TargetScan, miRTarBase and miRDB database analyses were applied to find the miRNA and target genes of lncRNA NONHSAT030515. A dual luciferase experiment was conducted to identify the direct target of NONHSAT030515. pcDNA3.1- NONHSAT030515 transfection and sh- NONHSAT030515 treatment were conducted to verify the role of lncRNA NONHSAT030515 in chondrogenic differentiation. The levels of Aggrecan, SOX9 and COL2A1 were analyzed by qRT-PCR and Western blot assay. Results Alcian blue staining, immunocytochemical, qRT-PCR, and Western blot have determined that lncRNA NONHSAT030515 can promote the chondrogenic differentiation of hADSCs. MiR-490- 5p was the direct target of NONHSAT030515, while BMPR2 was the target gene. This result was confirmed by luciferase reporter assay. Up-regulation of NONHSAT030515 promoted BMPR2 protein expression and promoted chondrogenic differentiation, whereas down-regulation of NONHSAT030515 caused completely opposite results. Conclusion LncRNA NONHSAT030515 promotes the chondrogenic differentiation of hADSCs through increasing BMPR2 expression by regulating miR-490- 5p.

Modalidades quimiossensoriais das larvas de Brycon orbignyanus / Chemosensory modalities of Brycon orbignyanus larvae

Com o objetivo de descrever o desenvolvimento das estruturas sensoriais quimiorreceptoras, corpúsculos gustativos e epitélio olfativo, das larvas de piracanjuba, Brycon orbignyanus, entre zero e 172 horas após eclosão, exemplares foram coletados a cada meia hora e fixados em fluído de bouin, por6 a 8 horas, e transferidos para solução de álcool 70%. As lâminas histológicas foram coradas por pelos métodos HE e Alcian Blue. As análises foram realizadas com fotomicroscópio de luz trinocular, acoplado a um microcomputador contendo um software de análise de imagens. Verificou-se que as larvas de piracanjuba, com 28 horas após a eclosão, desenvolveram estruturas quimiossensoriais importantes, como epitélio sensorial na cavidade olfatória e corpúsculos gustativos projetados na cavidade bucofaringiana e nas valvas orais, sugerindo que a quimiorrecepção é requerida na primeira alimentação exógena, que acontece neste momento. às 72 horas após eclosão, verificou-se a cavidade olfatória ampla e funcional e aumento de corpúsculos gustativos na cavidade bucofaringiana, o que revela uma elevada sensibilidade gustatória que, combinada com comportamento alimentar ativo, típico da espécie, sugere habilidade para detectar e selecionar alimentos e ingerí-los ou rejeitá-los, além de reconhecer substâncias tóxicas e venenosas. As larvas de piracanjuba, com 172 horas após eclosão, no final da metamorfose, apresentaram estruturas quimiossensoriais desenvolvidas, lamelas olfatórias formadas e gustação desenvolvida.

The clinico-pathologic profile of primary and recurrent orbital/periorbital plexiform neurofibromas (OPPN)

To evaluate and compare the clinical and histopathological profile of primary and recurrent orbital-periorbital plexiform neurofibromas (OPPN) in patients with neurofibromatosis type 1. We retrospectively evaluated 43 primary or recurrent neurofibroma (NF) specimens from 26 patients (2002 to 2018) at the King Khaled Eye Specialist Hospital, Saudi Arabia. Demographics, clinical presentation, and surgical intervention data were collected. Histopathological specimens were studied with hematoxylin-eosin, Alcian blue, and immunohistochemical markers; S-100, CD44, CD117, smooth muscle actin (SMA), neurofilament, and Ki-67. Of the 43 NFs specimens, 20 were primary and 23 recurrent tumors. For primary NF, the ratio of plexiform to the diffuse type was 13:7, however in recurrent tumors was 3:8 after the first recurrence, and 1:5 after multiple recurrences. Of the 17 patients with primary tumors that had paired recurrent tumors, 12/17 (70.6%) primary NFs were plexiform and 5/17 (29.4%) were diffuse. However, when tumors recurred, 13/17 tumors (76.5%) were diffuse and only 4/17 tumors (23.5%) had a plexiform pattern. The odds of a tumor having a diffuse pattern in recurrent NF was significantly higher than the plexiform pattern [OR = 7.8 (95% confidence interval 1.69:36.1) P = 0.008]. Primary plexiform NFs underwent an excision at a significantly younger age than the diffuse type. Recurrent NFs had significantly higher CD44, CD117, and neurofilament labeling (P = 0.02, P = 0.01 and P<0.001 respectively) but had significantly decreased Alcian blue, and S-100 labeling (P = 0.03, and P = 0.02 respectively) compared to primary tumors. SMA and Ki-67 proliferation index were not different between primary and recurrent NFs (P = 0.86, and P = 0.3 respectively). There appears to be a high risk for primary plexiform NFs to develop a diffuse histologic pattern when they recur. Immunohistochemical staining suggests a role of mast cells (CD117) and expression of infiltration makers (CD44) in the transformation of plexiform tumors to the diffuse phenotype.

Papillary Renal Cell Carcinoma with Extensive Spindle Cell Foci: Mimicker of Mucinous Tubular and Spindle Cell Carcinoma

Introduction/Objective Papillary Renal Cell Carcinoma (PRCC), the 2nd most common RCC, accounts for 10-15% of cases and is usually composed of tubules and papillae with foamy histiocytes in papillary cores. Mucinous tubular and spindle cell carcinoma (MTSC) is composed of tightly packed, elongated, curvilinear tubules with smooth luminal surfaces, separated by mucinous stroma. MTSC is associated with a more favorable prognosis than PRCC. PRCC and MTSC have significant histologic and histochemical overlap including elongated tubules and stromal Alcian blue positive mucin deposits. Methods/Case Report We report a case of a 75 year old female who underwent a robotic assisted partial nephrectomy for resection of a 5.7 x 5.2 x 5.0 cm left upper pole solid renal mass. Spindle cell change with elongated tubules reminiscent of MTSC was present in several blocks; however, the luminal surface was shaggy favoring PRCC. Patchy prominent extracellular Alcian blue positive mucin deposits were also present. PRCC and MTSC both express CK7, AMACR, and EMA. However, absent expression of E-cadherin and strong CD10 expression favored PRCC. Multiple foci of solid spindle cells in a whorled pattern with clear cell change, necrosis, and high grade nuclei bordering on sarcomatoid RCC were present in other blocks. Multiple papillations and psammoma bodies also supported PRCC. A spectrum of spindle cell change was present, ranging from elongated tubules reminiscent of MTSC to whorled foci with high grade nuclei approaching sarcomatoid RCC. Results (if a Case Study enter NA) NA Conclusion Submission of multiple sections and awareness of the protean morphologic features of PRCC are essential in making the correct diagnosis.

Prophylactic Efficacy of Tibial Plateau Levelling Osteotomy for a Canine Model with Experimentally Induced Degeneration of the Cranial Cruciate Ligament

Objective The aim of this study was to clarify the histological effects of tibial plateau levelling osteotomy on cranial cruciate ligament degeneration induced by excessive tibial plateau angle. Study Design Five female Beagles were used to bilaterally create excessive tibial plateau angle models surgically. A second tibial plateau levelling osteotomy was performed 11 months after the first surgery on the right stifle (tibial plateau levelling osteotomy group), and a sham operation that did not change the tibial plateau angle was performed on the left stifle (excessive tibial plateau angle group). At 6 months after the second surgery, the dogs were euthanatized. The cranial cruciate ligament was stained with haematoxylin–eosin to assess the cell density, Alcian-Blue to assess proteoglycans and Elastica-Eosin to assess elastic fibres, and immunohistochemically stained to assess type I (COL1) and type II collagen and SRY-type HMG box 9 (SOX9) expression. Results In each group, the cranial cruciate ligament degeneration, especially on the tibial side, including the presence of Alcian-Blue- and Elastica-Eosin-positive regions, decreased in COL1-positive regions, and enhancement of SOX9 expression was observed. Besides, compared with the tibial plateau levelling osteotomy group, the excessive tibial plateau angle group showed increases in Alcian-Blue- and Elastica-Eosin-positive regions and a decrease in the COL1-positive regions. Conclusion The results suggested that excessive tibial plateau angle-induced cranial cruciate ligament degeneration can be suppressed by reducing the biomechanical load on the cranial cruciate ligament by performing tibial plateau levelling osteotomy.

Construction of Loose Positively Charged NF Membrane by Layer-by-Layer Grafting of Polyphenol and Polyethyleneimine on the PES/Fe Substrate for Dye/Salt Separation

The effective separation of dyes and inorganic salts is highly desirable for recycling inorganic salts and water resource in printing and dyeing wastewater treatment. In this work, tannic acid (TA) and polyethyleneimine (PEI) were grafted on the PES/Fe ultrafiltration membrane via the coordination assembly and Michael addition strategy to fabricated a loose nanofiltration membrane (LNM). The effect of PEI concentration on membrane morphologies and properties was systematically investigated. The membrane surface becomes more hydrophilic and transforms into positive charge after the PEI grafting. The optimized PES/Fe-TA-PEI membrane possesses high pure water flux (124.6 L·m−2·h−1) and excellent dye rejections (98.5%, 99.8%, 98.4%, and 86.4% for Congo red, Eriochrome black T, Alcian blue 8GX, and Bromophenol blue, respectively) under 2 bar operation pressure. Meanwhile, the LNM showed a high Alcian blue 8GX rejection (>98.4%) and low NaCl rejection (<5.3%) for the dye/salt mixed solutions separation. Moreover, the PES/Fe-TA-PEI LNM exhibited good antifouling performance and long-term performance stability. These results reveal that such LNM shows great potential for effective fractionation of dyes and salts and recycling of textile wastewater.