Consolidation Tumor Ratio Combined With Pathological Features Could Predict Status of Lymph Nodes of Early-Stage Lung Adenocarcinoma

IntroductionStage IA lung adenocarcinoma manifested as part-solid nodules (PSNs), has attracted immense attention owing to its unique characteristics and the definition of its invasiveness remains unclear. We sought to develop a nomogram for predicting the status of lymph nodes of this kind of nodules.MethodsA total of 2,504 patients between September 2018 to October 2020 with part-solid nodules in our center were reviewed. Their histopathological features were extracted from paraffin sections, whereas frozen sections were reviewed to confirm the consistency of frozen sections and paraffin sections. Univariate and multivariate logistic regression analyses and Akaike information criterion (AIC) variable selection were performed to assess the risk factors of lymph node metastasis and construct the nomogram. The nomogram was subjected to bootstrap internal validation and external validation. The concordance index (C-index) was applied to evaluate the predictive accuracy and discriminative ability.ResultsWe enrolled 215 and 161 eligible patients in the training cohort and validation cohort, respectively. The sensitivity between frozen and paraffin sections on the presence of micropapillary/solid subtype was 78.4%. Multivariable analysis demonstrated that MVI, the presence of micropapillary/solid subtype, and CTR >0.61 were independently associated with lymph node metastasis (p < 0.01). Five risk factors were integrated into the nomogram. The nomogram demonstrated good accuracy in estimating the risk of lymph node metastasis, with a C-index of 0.945 (95% CI: 0.916–0.974) in the training cohort and a C-index of 0.975 (95% CI: 0.954–0.995) in the validation cohort. The model’s calibration was excellent in both cohorts.ConclusionThe nomogram established showed excellent discrimination and calibration and could predict the status of lymph nodes for patients with ≤3 cm PSNs. Also, this prediction model has the prediction potential before the end of surgery.

Mercury is present in neurons and oligodendrocytes in regions of the brain affected by Parkinson’s disease and co-localises with Lewy bodies

Objective Environmental toxicants are suspected to play a part in the pathogenesis of idiopathic Parkinson’s disease (PD) and may underlie its increasing incidence. Mercury exposure in humans is common and is increasing due to accelerating levels of atmospheric mercury, and mercury damages cells via oxidative stress, cell membrane damage, and autoimmunity, mechanisms suspected in the pathogenesis of PD. We therefore compared the cellular distribution of mercury in the tissues of people with and without PD who had evidence of previous mercury exposure by mercury being present in their locus ceruleus neurons. Materials and methods Paraffin sections from the brain and general organs of two people with PD, two people without PD with a history of mercury exposure, and ten people without PD or known mercury exposure, were stained for inorganic mercury using autometallography, combined with immunostaining for a-synuclein and glial cells. All had mercury-containing neurons in locus ceruleus neurons. Laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) was used to confirm the presence of mercury and to look for other potentially toxic elements. Autometallography-stained locus ceruleus paraffin sections were examined to compare the frequency of previous mercury exposure between 20 PD and 40 non-PD individuals. Results In PD brains, autometallography-detected mercury was seen in neurons affected by the disease, such as those in the substantia nigra, motor cortex, striatum, thalamus, and cerebellum. Mercury was seen in oligodendrocytes in white and grey matter. Mercury often co-localised with Lewy bodies and neurites. A more restricted distribution of brain mercury was seen in people without PD (both with or without known mercury exposure), with no mercury present in the substantia nigra, striatum, or thalamus. The presence of autometallography-detected mercury in PD was confirmed with LA-ICP-MS, which demonstrated other potentially toxic metals in the locus ceruleus and high iron levels in white matter. Autometallography-detected mercury was found in locus ceruleus neurons in a similar proportion of PD (65%) and non-PD (63%) individuals. Conclusions In people with PD, mercury was found in neurons and oligodendrocytes in regions of the brain that are affected by the disease, and often co-localised with aggregated a-synuclein. Mercury in the motor cortex, thalamus and striatum could result in bradykinesia and rigidity, and mercury in the cerebellum could cause tremor. People without PD had a restricted uptake of mercury into the brain. The similar frequency of mercury in the locus ceruleus of people with and without PD suggests these two groups have had comparable previous mercury exposures but that PD brains have a greater predisposition to take up circulating mercury. While this post mortem study does not provide a direct link between mercury and idiopathic PD, it adds to the body of evidence that metal toxicants such as mercury play a role in the disease. A precautionary approach would be to reduce rising mercury levels in the atmosphere by limiting the burning of fossil fuels, which may be contributing to the increasing incidence of PD.

Automated Hematoxylin and Eosin Staining with Leica AutoStainer XL v1

PURPOSE: To stain formalin fixed paraffin tissue for microscopic evaluation. Hematoxylin will stain the nuclei of the tissue blue/purple and Eosin will stain cytoplasmic elements a spectrum of brilliant pink. QUALITY CONTROL: Fixation: 10% Neutral Buffered Formalin Technique: Paraffin sections at 5 microns Control: PCRL PROCEDURE: Station #Time in StationExact Time?Solution Name185:00NoXylene175:00NoXylene165:00NoXylene150:20No100% Alcohol140:20No100% Alcohol130:20No95% Alcohol120:20No70% AlcoholWash 11:20NoWater83:30YesHematoxylinWash 20:30YesWaterWash 30:30YesWater90:10YesAcid AlcoholWash 40:30YesWater100:10YesAmmonia WaterWash 50:30YesWater112:00Yes95%70:20YesEosin60:30Yes95%50:30Yes100%40:30Yes100%30:30NoXylene21:00NoXylene11:00NoXylene RESULTS: Nuclei……………………………………………..……………Blue/Purple Cytoplasmic Elements………………………………….Shades of brilliant pink For digital storage and possible pathology review slides are scanned on the Leica Aperio slide scanner. The file output is .svs and can be viewed by downloading freely available software from Leica.

Common Transcriptomic Effects of Abatacept and Other DMARDs on Rheumatoid Arthritis Synovial Tissue

ObjectivesOur goal was to assess for the histological and transcriptomic effects of abatacept on RA synovia, and to compare them with previously published data from four other DMARDs: tocilizumab, rituximab, methotrexate, and adalimumab.MethodsSynovial tissue was obtained using ultrasound-guided biopsy from affected joints of 14 patients, before and 16 weeks after treatment with subcutaneous abatacept 125 mg weekly. Paraffin-sections were stained and scored for CD3+, CD20+, and CD68+ cell infiltration. Transcriptional profiling was performed using GeneChip Human Genome U133 Plus 2.0 arrays (Affymetrix) and analyzed on Genespring GX (Agilent). Pathway analyses were performed on Genespring GX, Metascape, and EnrichR.ResultsGene expression analysis identified 304 transcripts modulated by abatacept in synovial tissue. Downregulated genes were significantly enriched for immune processes, strongly overlapping with our findings on other therapies. Data were pooled across these studies, revealing that genes downregulated by DMARDs are significantly enriched for both T-cell and myeloid leukocyte activation pathways. Interestingly, DMARDs seem to have coordinate effects on the two pathways, with a stronger impact in good responders to therapy as compared to moderate and non-responders.ConclusionWe provide evidence that the effects of five DMARDs on the RA synovium culminate in the same pathways. This confirms previous studies suggesting the existence of common mediators downstream of DMARDs, independent of their primary targets.

Restorative Processes in the Thyroid Gland of Russian Long-Haired Rams in Postnatal Ontogenesis

The aimof this study is to study the recovery processes in the thyroid gland of Russian long-haired sheep in postnatal ontogenesis.Material and methods. The material for the study was 77 rams of the Russian long-haired breed of various ages. Fragments of the thyroid gland were fixed in a 10% solution of neutral formalin, in liquids of Shtive and Buena. Thin paraffin sections with a thickness of 4–5 μm were stained with hematoxylin and eosin, azan according to Heydenhein, and the PAS-reaction was applied. The average area of all follicles in the field of view was determined at a magnification of ×80 (ob. 10; oc. 8) using G.G. Autandilov ocular grid (50 fields of viewfor each animal). Based on the obtained absolute values of the area of the follicles and the interfollicular epithelium (mm 2 ), the relative value (%) of the area of the follicles and the interfollicular epithelium was determined. Using a screw eyepiece micrometer, the diameter of the nuclei of thyrocytes in the follicles and thyrocytes of the interfollicular epithelium was measured. The area of the nuclei was determined by the diameter of the nuclei. The mitotic index (MI) in ‰ was calculated . The number of dividing cells per thousand thyrocytes was calculated. At least 20,000 cells were counted for each animal.Results.It was found that in rams aged 15 days, the area of interfollicular epithelium was 86.5%, microfollicles occupied an area of 13.5%. Dividing cells were common, with the MI of follicular and interfollicular thyrocytes being 0.19‰. In rams at the age of 2 months, the number of small follicles increased, the areaof which was 19.7%. MI in follicles increased to 0.21‰, in interfollicular epithelium it decreased to 0.16‰. In rams at the age of 4 months, the area of follicles almost doubled, the area of interfollicular epithelium decreased by 16.1%. In rams aged 2–3 years, the area of interfollicular epithelium decreased, while MI decreased and was 0.13‰ in 3-year-old animals. In rams at the age of 4 and 5 years, there was a decrease in the functional activity of thefollicles. MI in large follicles was higher than in small follicles (0.12–0.13‰ versus 0.09‰).

AKR1B1 and AKR1B10 as Prognostic Biomarkers of Endometrioid Endometrial Carcinomas

The roles of aldo-keto reductase family 1 member B1 (AKR1B1) and B10 (AKR1B10) in the pathogenesis of many cancers have been widely reported but only briefly studied in endometrial cancer. To clarify the potential of AKR1B1 and AKR1B10 as tissue biomarkers of endometrial cancer, we evaluated the immunohistochemical levels of AKR1B1 and AKR1B10 in tissue paraffin sections from 101 well-characterized patients with endometrioid endometrial cancer and 12 patients with serous endometrial cancer and compared them with the clinicopathological data. Significantly higher immunohistochemical levels of AKR1B1 and AKR1B10 were found in adjacent non-neoplastic endometrial tissue compared to endometrioid endometrial cancer. A trend for better survival was observed in patients with higher immunohistochemical AKR1B1 and AKR1B10 levels. However, no statistically significant differences in overall survival or disease-free survival were observed when AKR1B1 or AKR1B10 were examined individually in endometrioid endometrial cancer. However, analysis of AKR1B1 and AKR1B10 together revealed significantly better overall and disease-free survival in patients with both AKR1B1 and AKR1B10 staining above the median values compared to all other patients. Multivariant Cox analysis identified strong AKR1B1 and AKR1B10 staining as a statistically important survival prediction factor. Conversely, no significant differences were found in serous endometrial cancer. Our results suggest that AKR1B1 and AKR1B10 play protective roles in endometrioid endometrial cancer and show potential as prognostic biomarkers.

Diagnosis of Ion-Exchange Resin Depositions in Paraffin Sections Using Corrective Light and Electron Microscopy-NanoSuit Method

Ion-exchange resins are commonly used to treat complications such as hyperkalemia, hyperphosphatemia, and hypercholesterolemia. Gastrointestinal complications may occur as side effects of such treatments. Sodium and calcium polystyrene sulfonate (PS-Ca) are cation-exchange resins comprising an insoluble structure that binds to potassium ions in the digestive tract and exchanges them with sodium and calcium ions, respectively, to promote their elimination. PS crystals are rhomboid, refractive, and basophilic in hematoxylin and eosin staining. To differentiate PS crystals from other ion-exchange resin crystals such as sevelamer and cholestyramine, periodic acid–Schiff, Ziehl–Neelsen, and Congo red staining are usually performed. Here, correlative light and electron microscopy (CLEM)-energy-dispersive X-ray spectroscopy and the NanoSuit method (CENM) was applied to perform a definitive identification of ion-exchange resins. CENM could detect sulfur in PS crystals without destroying the glass slides. Notably, PS retained its ion-exchange ability to bind potassium in paraffin sections. Differential diagnosis of anion-exchange resins, such as sevelamer and cholestyramine, was possible using these characteristics. The phosphorus:carbon ratio was higher in sevelamer than in cholestyramine after soaking paraffin sections in a phosphate solution. Therefore, CENM may be used for the differential pathological diagnosis of ion-exchange resins in paraffin sections.