Vascular architectural patterns in clear cell renal cell carcinoma and clear cell papillary renal cell carcinoma

AUTHORS

Sofia Canete-Portillo, Maria del Carmen Rodriguez Pena, Dezhi Wang, Diego F. Sanchez, George J. Netto & Cristina Magi-Galluzzi

ABSTRACT

Renal cell carcinomas (RCC) are well-vascularized tumors. Although clear cell RCC (CCRCC) show a characteristic vascular network, some cases show overlapping features with other RCC. We aimed to evaluate vascular architectural patterns, microvessel density (MVD), and endothelial cell density (ECD) in CCRCC compared to clear cell papillary RCC (ccpRCC). Thirty-four RCC (17 CCRCC and 17 ccpRCC) were included in the study. CD34 was used to evaluate vascular architectural patterns by microscopic estimation in all cases. CD34, ERG, and Bioquant Osteo 2019 Imaging Analysis Software were used to evaluate MVD and ECD in 17 CCRCC and 15 ccpRCC. Mean MVD was 526.63 in CCRCC vs. 426.18 in ccpRCC (p = 0.16); mean ECD was 937.50 in CCRCC vs. 1060.21 in ccpRCC (p = 0.25). CD34 highlighted four distinct vascular architectural patterns: pseudoacinar, Golgi-like, lacunae, and scattered. Lacunae and pseudoacinar was the most frequent combination in CCRCC; lacunae and Golgi-like was the predominant combination among ccpRCC. Pseudoacinar was most extensive in CCRCC and least in ccpRCC; Golgi-like was predominant in ccpRCC and uncommon in CCRCC. The extent of pseudoacinar and Golgi-like vascular architectural patterns was significantly different between CCRCC and ccpRCC (p < 0.05). Pathologists acquainted with these different vascular architectural patterns may utilize them as an additional tool in the distinction of CCRCC from ccpRCC.

Mechanism of Pingyang Jiangya Formula in treating hypertension based on network pharmacology and in vivo study

AUTHORS

Liu Deguo, Li Zirong, Chen Qihua, Wang Yuhong, Xiao Changjiang

ABSTRACT

Objective

This study aimed to analyze the mechanism of action of the Pingyang Jiangya Formula (平阳降压方, PYJYF) in treating hypertension, based on network pharmacology, and to verify the subsequent predictions through animal experiments.

Methods

The active components and related target genes of PYJYF were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM), Encyclopedia of Traditional Chinese Medicine (ETCM), and DrugBank databases and available literature. The hypertension target genes were screened based on Therapeutic Target Database (TTD), GeneCards, Online Mendelian Inheritance in Man (OMIM), UniProt, and relevant literature. The component-disease-target network intersection target genes were inputted into the STRING database, and the key target genes were selected according to the degree algorithm. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed to explore the multitarget mechanism of action and molecular regulatory network of PYJYF in the treatment of hypertension. To verify this prediction, we used PYJYF to intervene in spontaneously hypertensive rats (SHRs) and Wistar–Kyoto rats (WKY) as normal control, and the noninvasive tail artery manometry method was used to measure systolic blood pressure (SBP) in the rat tail before PYJYF intervention. After drug intervention, the SBP of each group rats were measured and compared every week. Enzyme-linked immunosorbent assay (ELISA) was used to test plasma renin, angiotensin II (Ang II), and aldosterone (Ald) levels, and hematoxylin-eosin (HE) staining was used to observe pathological damage to the renal vessels in each group of rats. Western blot and reverse transcription real-time quantitative PCR (RT-PCR) were used to detect the protein and mRNA expression levels of PI3K, AKT1, BAX, and Bcl-2, respectively.

Results

A total of 4 123 hypertension targets were obtained from related databases. From the TCMSP and chemical databases, 78 active components of PYJYF and the corresponding 401 drug targets were retrieved. Data analysis revealed that 208 drug targets directly interacted with the hypertension targets in PYJYF. The 10 targets most closely related to hypertension target proteins in PYJYF were directly retrieved from relevant databases. GO analysis revealed that 10 direct target proteins were involved in all aspects of the antihypertensive effects of PYJYF, as well as molecular biological processes, such as the regulation of blood pressure, renin-angiotensin-aldosterone system (RAAS), angiotensin-mediated ligand reactions, and biological stimulation of cardiomyocyte apoptosis. KEGG pathway enrichment analysis revealed that PYJYF directly affected 20 signaling pathways associated with hypertension. In animal experiments, PYJYF reduced the protein and mRNA levels of PI3K, Akt, and Bax and upregulated the expression of the protein and mRNA levels of Bcl-2, reduced plasma renin, Ang II, and Ald levels, improved the hyperactivity of RAAS, and significantly reduced SBP in SHRs.

Conclusion

PYJYF is effective for hypertension therapy that acts through multiple compounds and targets. The possible underlying molecular mechanism includes regulating the PI3K/Akt signaling pathway to suppress RAAS, increasing the ratio of Bcl-2/Bax proteins, and inhibiting apoptosis, thereby mediating the repair of renal and renal vascular damage caused by hypertension. These findings warrant further research for use in clinical settings.

Evaluation of osteogenic potential of Cissus quadrangularis on mandibular alveolar ridge distraction

AUTHORS

Alaa Abdelqader Altaweel, Abdel Aziz Baiomy Abdullah Baiomy, Hazem Shawky Shoshan, Hisham Abbas, Ahmed Abdel-Shakour Abdel-Hafiz, Abd El-Hamid Gaber, Amr Abdelfatah Zewail & Marwa A. M. Elshiekh

ABSTRACT

Background

This randomized clinical trial was designed to evaluate osteogenic potential of Cissus quadrangularis in alveolar distraction to facilitate implant installation.

Material and methods

Twenty patients with atrophic ridge were treated by alveolar distraction. After completing distractor activation, patients were randomly divided into two equal groups according to administered drug (placebo and Cissus quadrangularis group). After a consolidation period, distractors were removed and implants were inserted. Clinical evaluation was done to assess wound healing, and distractor and implant stability. Histological evaluation was performed at time of implant insertion. Radiographic evaluation was performed to assess bone volume and density after distraction, as well as, density and bone loss around implant.

Results

Radiographic and histological results showed that bone formation and maturation of study group were faster than that of control group. There was a significant increased bone density in distracted area and around implant in study group than control group. A significant bone loss at end of consolidation period, and around implant at end of the study was reported in control group than study group.

Conclusion

Cissus quadrangularis administration during the consolidation period is associated with increased osteogenic potential of distracted bone. The histological and radiographic findings of current study proved that Cissus quadrangularis not only enhances rate of new bone formation, but also bone density to withstand the biomechanical requirements of implant placement in a shorter time.

Osteoclast-mediated bone loss observed in a COVID-19 mouse model

AUTHORS

Olatundun D. Awosanya, Christopher E. Dalloul, Rachel J. Blosser, Ushashi C. Dadwal, Mariel Carozza, Karen Boschen, Michael J. Klemsz, Nancy A. Johnston, Angela Bruzzaniti, Christopher M. Robinson, Edward F. Srour, Melissa A. Kacena

ABSTRACT

The consequences of SARS-CoV-2 infection on the musculoskeletal system represent a dangerous knowledge gap. Aging patients are at added risk for SARS-CoV-2 infection; therefore, a greater understanding of the resulting musculoskeletal sequelae of SARS-CoV-2 infection may help guide clinical strategies. This study examined fundamental bone parameters among mice treated with escalating viral loads. Male C57BL/6J (WT, n = 17) and B6.Cg-Tg(K18-ACE2)2Prlmn/J mice (K18-hACE2 transgenic mice, n = 21) expressing human ACE2 (TG) were divided into eight groups (n = 4–6/group) and subjected to intranasal dosing of 0, 1 × 103, 1 × 104, and 1 × 105 PFU (plaque forming units) of human SARS-CoV-2. Animal health was assessed daily by veterinary staff using established and validated scoring criteria (activity, posture, body condition scores and body weight). We report here that mock and WT infected mice were healthy and completed the study, surviving until 12–14 days post infection (dpi). In contrast, the TG mice infected with 1 × 105 PFU all experienced severe health declines that necessitated early euthanasia (6–7 dpi). For TG mice infected with 1 × 104 PFU, 2 mice were also euthanized after 7 dpi, while 3 mice showed signs of moderate disease at day 6 dpi, but recovered fully by day 11 dpi. Four of the 5 TG mice that were infected with 1 × 103 PFU remained healthy throughout the study. This suggests that our study mimics what is seen during human disease, where some patients develop severe disease resulting in death, while others have moderate to severe disease but recover, and others are asymptomatic. At necropsy, femurs were extracted and analyzed by μCT. No difference was found in μCT determined bone parameters among the WT groups. There was, however, a significant 24.4% decrease in trabecular bone volume fraction (p = 0.0009), 19.0% decrease in trabecular number (p = 0.004), 6.2% decrease in trabecular thickness (p = 0.04), and a 9.8% increase in trabecular separation (p = 0.04) among surviving TG mice receiving any viral load compared to non-infected controls. No differences in cortical bone parameters were detected. TRAP staining revealed surviving infected mice had a significant 64% increase in osteoclast number, a 27% increase in osteoclast surface, and a 38% increase in osteoclasts per bone surface. While more studies are needed to investigate the long-term consequences of SARS-CoV-2 infection on skeletal health, this study demonstrates a significant reduction in several bone parameters and corresponding robust increases in osteoclast number observed within 2 weeks post-infection in surviving asymptomatic and moderately affected mice.

Development of Nystagmus With the Absence of MYOD Expression in the Extraocular Muscles

AUTHORS

Laura L. Johnson; Rachel B. Kueppers; Erin Y. Shen; Jolene C. Rudell; Linda K. McLoon

ABSTRACT

Purpose: Myoblast determination protein 1 (MYOD) is a critical myogenic regulatory factor in muscle development, differentiation, myofiber repair, and regeneration. As the extraocular muscles significantly remodel their myofibers throughout life compared with limb skeletal muscles, we hypothesized that the absence of MYOD would result in their abnormal structure and function. To assess structural and functional changes in the extraocular muscles in MyoD−/− mice, fiber size and number and optokinetic nystagmus reflex (OKN) responses were examined.

Methods: OKN was measured in MyoD−/− mice and littermate wild-type controls at 3, 6, and 12 months. The extraocular muscles were examined histologically for changes in mean myofiber cross-sectional area, total myofiber number, and nuclei immunostained for PAX7 and PITX2, markers of myogenic precursor cells.

Results: The MyoD−/− mice developed nystagmus, with both jerk and pendular waveforms, in the absence and in the presence of moving visual stimulation. At 12 months, there were significant losses in mean myofiber cross-sectional area and in total number of orbital layer fibers in all rectus muscles, as well as in global layer fibers in the superior and inferior rectus muscles. Haploinsufficient mice showed abnormal OKN responses. PITX2-positive cell entry into myofibers of the MyoD−/− mice was significantly reduced.

Conclusions: This study is the first demonstration of the development of nystagmus in the constitutive absence of expression of the muscle-specific transcription factor MYOD. We hypothesize that myofiber loss over time may alter anterograde and/or retrograde communication between the motor nerves and extraocular muscles that are critical for maintaining normalcy of extraocular muscle function.

Differences in the Microarchitectural and Histomorphologic Characteristics Between Glucocorticoid-induced Osteonecrosis of Femoral Head and Alcohol-induced Osteonecrosis of Femoral Head

AUTHORS

Yiwei Chen, Kexin Liu, Yu Miao, Bin Zhu, Feng Xue, Junhui Yin, Minghao Zheng, guangyi li, Changqing Zhang

ABSTRACT

Aims

To analyze microarchitecture and histomorphology characteristics of different regions in femoral heads from patients with glucocorticoid-induced osteonecrosis of femoral head (GIONFH) and alcohol-induced osteonecrosis of femoral head (AIONFH).

Methods

Patients diagnosed with GIONFH and AIONFH were recruited. Femoral heads were obtained after total hip replacement. Micro-CT was applied to evaluate the microstructure of 9 regions of interest (ROIs) in the femoral head. Along the supero-inferior orientation, the femoral head was divided into necrotic region, reactive interface, and normal region; along the medio-lateral orientation, the femoral head was divided into medial region, central region and lateral region. Decalcified and undecalcified bone histology were then performed to assess histopathological alterations and bone remodeling levels.

Results

42 GIONFH patients (50 hips) and 43 AIONFH patients (50 hips) anticipated in the study. In the necrotic region, most of the microarchitectural parameters did not differ significantly between GIONFH and AIONFH, whereas both the reactive interface and normal region illustrated significant differences in the microstructure and histomorphometry. The reactive interface and normal region exhibited a less sclerotic microarchitecture, but a higher bone remodeling level in GIONFH as compared with AIONFH. Despite similar necrotic pathological manifestations, subchondral trabecular microfracture in the necrotic region was more severe and vasculature of the reactive interface was more abundant in GIONFH.

Conclusions

Although these two subtypes of ONFH shared similar microarchitecture and pathological features in the necrotic region, GIONFH exhibited a less sclerotic microarchitecture and a more active bone metabolic status in both the reactive interface and normal region.