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Objective: In the era of image-guided adaptive radiotherapy, definition of the clinical target volume (CTV) is a challenge in various solid tumors, including esophageal cancer (EC). Many tumor microenvironmental factors, e.g., tumor cell proliferation or cancer stem cells, are hypothesized to be involved in microscopic tumor extension (MTE). Therefore, this study assessed the expression of FAK, ILK, CD44, HIF-1α, and Ki67 in EC patients after neoadjuvant radiochemotherapy followed by tumor resection (NRCHT+R) and correlated these markers with the MTE. Methods: Formalin-fixed paraffin-embedded tumor resection specimens of ten EC patients were analyzed using multiplex immunofluorescence staining. Since gold fiducial markers had been endoscopically implanted at the proximal and distal tumor borders prior to NRCHT+R, correlation of the markers with the MTE was feasible. Results: In tumor resection specimens of EC patients, the overall percentages of FAK+, CD44+, HIF-1α+, and Ki67+ cells were higher in tumor nests than in the tumor stroma, with the outcome for Ki67+ cells reaching statistical significance (p < 0.001). Conversely, expression of ILK+ cells was higher in tumor stroma, albeit not statistically significantly. In three patients, MTE beyond the fiducial markers was found, reaching up to 31 mm. Conclusion: Our findings indicate that the overall expression of FAK, HIF-1α, Ki67, and CD44 was higher in tumor nests, whereas that of ILK was higher in tumor stroma. Differences in the TME between patients with residual tumor cells in the original CTV compared to those without were not found. Thus, there is insufficient evidence that the TME influences the required CTV margin on an individual patient basis. Trial registration number and date: BO-EK-148042017 and BO-EK-177042022 on 20.06.2022, DRKS00011886, https://drks.de/search/de/trial/DRKS00011886. [ABSTRACT FROM AUTHOR]

The immunohistochemical assessment of mismatch repair (MMR) proteins represents a pivotal screening tool for identifying Lynch syndrome (LS)-related cancers, as the loss of their expression often indicates MMR dysfunction associated with genetic or epigenetic alterations. Frequently, LS-related colorectal cancers present germline pathogenic variants in the MLH1 or MSH2 genes, which result in the simultaneous immunohistochemical loss of MLH1 and PMS2 or MSH2 and MSH6 proteins expression, respectively. Less commonly observed is the single involvement of the MSH6 or PMS2 proteins expression, indicative of the presence of germline pathogenic variants in the corresponding genes. Extremely rarely reported are the null immunohistochemistry phenotypes represented by the complete loss of expression of all MMR proteins. The molecular mechanisms contributing to the raising of this latter uncommon immunohistochemical phenotype are derived from the combination of pathogenic germline variants in MMR genes with the somatic hypermethylation of the MLH1 gene promoter. This study focuses on elucidating the molecular cascade leading to the development of the null immunohistochemical phenotype, providing valuable insights into understanding the sequential molecular events driving the LS-associated tumorigenesis, which may have pivotal implications in the clinical management of patients with LS-related cancers. [ABSTRACT FROM AUTHOR]

Purpose: The main purpose of this study was to perform an immunohistochemical, functional, and anatomical evaluation of patients with idiopathic epiretinal membrane (ERM). Methods: Twenty-four specimens of idiopathic ERM from 24 consecutive patients who underwent 23 G pars plana vitrectomy for ERM and internal limiting membrane (ILM) peeling at the San Juan University Hospital in Alicante (Spain) in 2019 were analyzed. All patients underwent a complete ophthalmological examination including measurement of best corrected visual acuity (BCVA) and macular analysis by spectral-domain optical coherence tomography (SD-OCT) at the time of diagnosis and 3 months after surgery. Specific glial fibrillar acid protein antibodies (GFAP) and S100 calcium-binding protein β (S100β) immunostaining markers were used to identify the macroglial component of the ERM, Müller cells, and astrocytes. Ionized calcium-binding adapter molecule 1 protein (Iba1) antibodies were used as specific markers for inflammatory cells, such as microglia and macrophages. Results: Mean preoperative BCVA measured with Snellen chart was 0.3 and 0.6 preoperatively and at 3 months after surgery, respectively. SD-OCT identified 15 patients (62.5%) with a disruption of the outer retinal hyperreflective bands. The immunohistochemical study showed the presence of Müller cells in almost all cases (91.6%), as well of abundant microglia and macrophages. Microglia and macrophages were more frequently present in earlier stages of ERM. Microglia were present in ERM independently of the outer retinal hyperreflective bands integrity as measured by SD-OCT. A greater presence of macrophages was found in those ERMs with no outer retinal hyperreflective band disruption. Conclusions: Müller cells seem to be the most frequent cell group in ERMs, with also presence of microglia cells and macrophages. Astrocytes were more frequently found in early stages of ERMs. Microglia and macrophages were most frequent in ERMs with early stage (1, 2, or 3) than in advanced stages (4). [ABSTRACT FROM AUTHOR]

Undifferentiated pleomorphic osteosarcoma belongs to the group of rarely occurring tumors. Despite the treatment, the disease progresses in 30–40 % of patients with osteosarcomas. The main route of metastasis of bone tissue sarcomas is hematogenous, while lymphogenic spread is observed less frequently. As a rule, secondary metastatic changes occur in the lungs. Less often there is a secondary lesion of the bones of the skeleton and brain. Metastatic lesion of uterus, fallopian tubes and ovaries in malignant undifferentiated pleomorphic sarcoma is extremely rare. Therefore, we found it interesting to describe a clinical case of such a rare metastatic lesion. Patient K. underwent amputation of the left limb at the level of the lower third of the femur for undifferentiated pleomorphic sarcoma of the left tibia in 2019, and 4 courses of adjuvant polychemotherapy were performed. In 20 months after completion of complex treatment of the primary tumor, complaints of lower abdominal pain, increased body temperature up to 37.8 °C in the evenings appeared. According to the results of follow-up examination, a voluminous, multinodular, solid mass of merging character was detected in the pelvis, with total dimensions of up to 11 cm, and a cavitary mass of up to 5 cm was detected in the posterior vault. A trepan-biopsy of the mass in the projection of the right ovary was performed. The morphological picture in the volume of trepan biopsy specimens is characteristic of spindle cell sarcoma. Metastasis of undifferentiated pleomorphic bone sarcoma (malignant fibrous histiocytoma) is most likely. Due to metastatic lesions of the uterus, fallopian tubes, ovaries, omentum, mesentery and serous membrane of the colon loops, peritoneum of the bladder, surgical intervention in the volume of removal of the distal part of the sigmoid colon, rectosigmoid, upper ampullary parts of the rectum, uterus with fallopian tubes and ovaries, appendix was performed. Immunohistochemical study of the postoperative material revealed that the immunophenotype of tumor cells confirmed the morphological picture typical for undifferentiated pleomorphic bone sarcoma. The patient was further prescribed antitumor drug therapy. This clinical case demonstrates a rare, atypical metastasis of undifferentiated pleomorphic osteosarcoma, which allows to expand the knowledge about the flow of malignant diseases of this localization.

The prevalence of thyroid cancer has shown a progressive rise over time. This study aimed to explore the expression and underlying mechanisms of decorin (DCN) in follicular thyroid carcinoma (FTC), employing bioinformatics analysis and immunohistochemistry techniques. The GSE27155 dataset was obtained from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) and core DEGs were identified through data mining and analysis using the R language and online databases. The expression of core DEGs was validated using The Cancer Genome Atlas database. Additionally, the correlation between DCN and clinicopathological stage, tumor-infiltrating lymphocytes, and hotspot molecules in thyroid cancer was assessed using the Gene Expression Profiling Interactive Analysis and TIMER databases. Immunohistochemical (IHC) analysis was then conducted to verify the differential expression of core DCN in FTC and adjacent tissues. We confirmed the downregulation of three DEGs (DCN, GPC3, and PDGFRA). Furthermore, the analysis revealed a significant association between DCN expression and the clinical stage of patients with thyroid cancer (p<0.0001). DCN expression and the infiltration of several immune cells were positively correlated (p<0.01). A significant positive correlation was also noted between DCN and the NRAS and KRAS genes (partial cor>0, p<0.05). Immunohistochemical analyses revealed a significantly lower staining score (3.071 ± 2.493) for DCN protein in cancer tissues than that in adjacent tissues (8.643 ± 2.094) (p<0.0001). DCN is underexpressed and contributes to tumor progression in FTC. Thus, DCN serves as a tumor suppressor gene in FTC and a promising therapeutic target. [ABSTRACT FROM AUTHOR]

Objectives: To determine the roles of small GTP-binding proteins Rac1, Rac2, and Rac3 expression in pterygial tissue and to compare these expressions with normal conjunctival tissue. Materials and Methods: Seventy-eight patients with primary pterygium were enrolled. Healthy conjunctival graft specimens obtained during pterygium surgery were used as control tissue. The real-time polymerase chain reaction method on the BioMark HD dynamic array system was utilized in genomic mRNA for the gene expression analysis. Protein expressions were analyzed using western blot and immunohistochemical methods. Results: RAC1, RAC2, and RAC3 gene expressions in pterygial tissues were not markedly elevated when compared to the control specimens (p>0.05). As a very low level of RAC1 gene expression was observed, further protein expression analysis was performed for the Rac2 and Rac3 proteins. Western blot and immunohistochemical analysis of Rac2 and Rac3 protein expression revealed no significant differences between pterygial and healthy tissues (p>0.05). Conclusion: This is the first study to identify the contribution of Rac proteins in pterygium. Our results indicate that the small GTP-binding protein Rac may not be involved in pterygium pathogenesis.

Aim: to present a clinical and morphological observation of an extremely rare combination of medullary carcinoma of the jejunum and intestinal lymphangiectasia in a 33-year-old patient with clinical features of malabsorption syndrome over the 10 years.Key points. An autopsy revealed a tumor formation spreading from the wall of the jejunum to the mesentery, with metastases to the mesenteric lymph nodes. The medullary carcinoma with positive expression of СD117, DOG1, EMA, PanCK, PDL-1, vimentin, mosaic non-intense expression of CA19-9, calretinin, CD10, CDX2, CEA, MUC-5AC, SATB2, and negative reaction to ALK, CD3, CD8, CD20, CD30, CD31, CD34, CD45, CD56, chromogranin, CK7, CK20, desmin. The proliferative index was high: Ki-67 > 80 %. Moreover, during the histological examination of the intestinal wall, intestinal lymphangiectasia complicated by the malabsorption syndrome was revealed.Conclusion. The uniqueness of this clinical and morphological case is in the combination of medullary carcinoma of the jejunum metastasized to the mesenteric lymph nodes with the underlying intestinal lymphangiectasia accompanied by the development of malabsorption syndrome.

The identification of markers for early diagnosis, prognosis, and improvement of therapeutic options represents an unmet clinical need to increase survival in Non-Small Cell Lung Cancer (NSCLC), a neoplasm still characterized by very high incidence and mortality. Here, we investigated whether proline dehydrogenase (PRODH), a mitochondrial flavoenzyme catalyzing the key step in proline degradation, played a role in NSCLC tumorigenesis. PRODH expression was investigated by immunohistochemistry; digital PCR, quantitative PCR, immunoblotting, measurement of reactive oxygen species (ROS), and functional cellular assays were carried out. PRODH expression was found in the majority of lung adenocarcinomas (ADCs). Patients with PRODH-positive tumors had better cancer-free specific and overall survival compared to those with negative tumors. Ectopic modulation of PRODH expression in NCI-H1299 and the other tested lung ADC cell lines decreased cell survival. Moreover, cell proliferation curves showed delayed growth in NCI-H1299, Calu-6 and A549 cell lines when PRODH-expressing clones were compared to control clones. The 3D growth in soft agar was also impaired in the presence of PRODH. PRODH increased reactive oxygen species production and induced cellular senescence in the NCI-H1299 cell line. This study supports a role of PRODH in decreasing survival and growth of lung ADC cells by inducing cellular senescence. [ABSTRACT FROM AUTHOR]

The identification of reliable brain-specific biomarkers in periphery contributes to better understanding of normal neurophysiology and neuropsychiatric diseases. The neurospecific proteins BDNF, NSE, VILIP-1, and S100B play an important role in the pathogenesis of neuropsychiatric disorders, including epilepsy. This study aimed to assess the correspondence of the expression of BDNF, NSE, VILIP-1, and S100B in the blood (serum and peripheral blood mononuclear cells (PBMCs)) to the in vivo hippocampal levels of subjects with drug-resistant epilepsy who underwent neurosurgery (N = 44) using multiplex solid-phase analysis, ELISA, and immunohistochemical methods, as well as to analyze the correlations and associations of the blood and hippocampal levels of these proteins with clinical parameters. We first studied the concordance between in vivo brain and blood levels of BDNF, NSE, VILIP-1, and S100B in epileptic patients. A positive correlation for NSE between hippocampal and PBMC levels was revealed. NSE levels in PBMCs were also significantly correlated with average seizure duration. BDNF levels in PBMCs were associated with seizure frequency and hippocampal sclerosis. Thus, NSE and BDNF levels in PBMCs may have potential as clinically significant biomarkers. Significant correlations between the levels of the neurospecific proteins studied herein suggest interactions between BDNF, NSE, VILIP-1, and S100B in the pathophysiology of epilepsy. [ABSTRACT FROM AUTHOR]

Objectives: To determine the roles of small GTP-binding proteins Rac1, Rac2, and Rac3 expression in pterygial tissue and to compare these expressions with normal conjunctival tissue. Materials and Methods: Seventy-eight patients with primary pterygium were enrolled. Healthy conjunctival graft specimens obtained during pterygium surgery were used as control tissue. The real-time polymerase chain reaction method on the BioMark HD dynamic array system was utilized in genomic mRNA for the gene expression analysis. Protein expressions were analyzed using western blot and immunohistochemical methods. Results: RAC1, RAC2, and RAC3 gene expressions in pterygial tissues were not markedly elevated when compared to the control specimens (p>0.05). As a very low level of RAC1 gene expression was observed, further protein expression analysis was performed for the Rac2 and Rac3 proteins. Western blot and immunohistochemical analysis of Rac2 and Rac3 protein expression revealed no significant differences between pterygial and healthy tissues (p>0.05). Conclusion: This is the first study to identify the contribution of Rac proteins in pterygium. Our results indicate that the small GTP-binding protein Rac may not be involved in pterygium pathogenesis. [ABSTRACT FROM AUTHOR]

Introduction. High-risk human papilloma virus (Hpv), especially genotype 16, causes oropharyngeal squamous cell carcinoma (OSCC). It is detected in about 70 % of tumors developing from lymphoid tissue of the tonsils or the base of the tongue. Due to the increased number of Hpv-positive OSCC, Hpv status is considered a marker of OSCC clinical outcome. Easy testing, low cost, reliability, and high sensitivity of immunohistochemical analysis for p16INk4a allowed to widely use this method for Hpv status determination.Aim. To determine the association between programmed death-ligand 1 (pD-L1) and p53 expression and presence of indirect Hpv marker – p16INk4a – in patients with OSCC.Materials and methods. The study included 76 patients with OSCC т1–4N0–3m0 who received treatment at the Republican Specialized Scientific and practical medical Center of Oncology and Radiology (n = 37) and its Tashkent branch (n = 39) between 2015 and 2020. for all selected patients, retrospective immunohistochemical analysis for the presence of p16INk4a, pD-L1 and р53 in tumor samples fixed with formalin in paraffin blocks was performed. In our work, immunohistochemical examination for p16INk4a was the only relevant tool for Hpv status determination. To reinforce its prognostic significance, we used additional molecular markers pD-L1 and p53 which play an important role in carcinogenic transformation and OSCC progression.Results. The results of immunohistochemical analysis showed that p16INk4a overexpression was accompanied by positive pD-L1 reaction in 46 % (6/13) of cases; there were no cases of positive expression of mutant type p53. wild type p53 was identified in only 1 (3 %) case in combination with p16INk4a overexpression.Conclusion. The developed panel consisting of 3 molecular markers (p16INk4a, pD-L1 and р53) may open new horizons in accurate prognosis, risk stratification and understanding of OSCC molecular signature. This, in turn, will help clinicians in selection of individual therapy strategies for treatment de-escalation and outcome optimization.

Introduction: Glucocorticosteroid therapy is the basic therapeutic option for rheumatoid arthritis; however, long-term treatment with low and medium doses is associated with the development of negative side effects. As reported, oxidative stress is crucial in the pathogenesis of rheumatoid arthritis. Therefore, pharmacotherapy that combines application of methylprednisolone pulse therapy and an aqueous molecular hydrogen solution seems to be a reasonable treatment option in these cases. Materials and Methods: The study of the effectiveness of combined pharmacotherapy for rheumatoid arthritis was carried out on mature male rats. Rheumatoid arthritis was simulated by introducing bovine type II collagen into the right knee joint. Animals of the 1st – control – group received placebo; animals of the 2nd group received molecular hydrogen-enriched water intragastrically; animals of the 3rd group received methylprednisolone solution intravenously by catheterization of the tail vein, and animals of the 4th group received water enriched with molecular hydrogenintragastrically and methylprednisolone solution intravenously. After withdrawal of animals from the experiment, microsections of their joint tissues were analysed histologically and biomarkers of the joint inflammation were detected immunohistochemically. Results and Discussion: Morphological analysis of microsections taken from animals of the 4th group evidenced effectiveness of the combined therapy based on quantitative estimation of the inflammatory biomarker expression. Dynamic polarization of M1/M2 macrophage was manifested with high quality in animals of this group. Conclusion: The search for new therapeutic options for rheumatoid arthritis is expanded due to major antioxidant substances that can reduce duration of treatment, while ensuring positive dynamics of the course of the disease.

Introduction. The adverse effects of urine on unadapted tissues are known. This is also entirely relevant for buccal grafts used in augmentation urethroplasty, where these effects have not been thoroughly studied so far.Objective. To assess the ongoing pathomorphosis in buccal grafts used for urethral augmentation of extensive strictures in the bulbous region and to evaluate how much urine influences their histological transformation following surgery.Materials & methods. The study included 15 patients with extensive strictures of the bulbous urethra, who underwent a two-stage augmentation urethroplasty with buccal grafts. The grafts pathomorphosis was studied 6 months after the first surgery stage where urethrotomy had been performed with graft augmentation of the dorsal semicircle and the formation of distal and proximal neomeatuses. Natural urination through the latter was restored on days 14 – 20 following the surgery. During the second stage, six months later, urethral tubularisation was performed with two preliminary biopsies of the proximal and distal segments of the grafts implanted in the areas of the neomeatuses formed earlier. The distal area of the graft had no contact with urine, while this contact has occurred in the proximal segment since restoration of natural urination. In biopsy specimens, pathomorphosis of the grafts was studied using immunohistochemical markers: vimentin, clone SRL33; CD34, clone QBEnd/10; MSA HHF-35; СD3, clone LN10; Bcl-2, clone bcl-2\100\D5; CK-HMW 34BE-12.Results. It was found that inflammation was minimal in areas of grafts implanted that had no contact with urine, while in areas where such contact occurred it was verified to be pronounced even 6 months after the operation. On the submucosal level, this was manifested by an uneven arrangement of collagen fibers, a dysplastically developed vascular network, uneven proliferation of the endothelium with swelling and loss of cellular connectivity, in contrast to areas where there was no contact with urine. In such areas, the graft submucosa had a dense collagen framework with organized microvasculature and uniform epithelial surface.Conclusion. The impact of urine on buccal grafts used in augmentation urethroplasty is characterised by the disorganisation of its collagen framework, with a pronounced inflammatory component and the “reactivity” of the epithelial lining to the “toxic agent” that persists even 6 months after surgery. This may underlie the risk of a stenosis relapse in the proximal anastomosis area.

In this study, in experiments on 35 healthy male mice of the C57BL/6 line weighing 25±3 g, it was shown that loud sound/music for 2 hours causes a temporary increase in the permeability of the blood-brain barrier (BBB) in rodents. To investigate changes in the permeability of the blood-brain barrier, the molecular mechanisms responsible for its discovery were investigated using laser speckle-contrast imaging of regional cerebral blood fl ow (rCBF), immunohistochemical analysis and biochemical analysis of adrenaline in blood plasma. With a sound-dependent increase in the permeability of the blood-brain barrier, there was a decrease in signal intensity from CLND-5, Occ, JAM and an increase in the signal from ZO-1. However, after 4 hours, the signal intensity from the studied proteins was restored, which may be due to their internalization. The results of the study of the eff ects of music and sound on of BBB in the intact brain require a revision of traditional knowledge about the barrier functions of the brain and open up new opportunities for non-invasive drug delivery strategies. They also may off er some insight into the etiology of brain disorders that follow inadvertent or deliberate exposure to very loud sounds, i.e. battle or rock concerts.

The immunohistochemical assessment of mismatch repair (MMR) proteins represents a pivotal screening tool for identifying Lynch syndrome (LS)-related cancers, as the loss of their expression often indicates MMR dysfunction associated with genetic or epigenetic alterations. Frequently, LS-related colorectal cancers present germline pathogenic variants in the MLH1 or MSH2 genes, which result in the simultaneous immunohistochemical loss of MLH1 and PMS2 or MSH2 and MSH6 proteins expression, respectively. Less commonly observed is the single involvement of the MSH6 or PMS2 proteins expression, indicative of the presence of germline pathogenic variants in the corresponding genes. Extremely rarely reported are the null immunohistochemistry phenotypes represented by the complete loss of expression of all MMR proteins. The molecular mechanisms contributing to the raising of this latter uncommon immunohistochemical phenotype are derived from the combination of pathogenic germline variants in MMR genes with the somatic hypermethylation of the MLH1 gene promoter. This study focuses on elucidating the molecular cascade leading to the development of the null immunohistochemical phenotype, providing valuable insights into understanding the sequential molecular events driving the LS-associated tumorigenesis, which may have pivotal implications in the clinical management of patients with LS-related cancers.

Metastasis to the gastrointestinal tract is rare. A 59-year-old woman who had a history of an invasive lobular carcinoma of breast with clinical complete response visited our hospital and complained of an upper abdominal pain and distension. We performed an upper gastrointestinal endoscopy which showed only a gastric ulcer without any malignant findings. She experienced a recurrence of symptoms 2 months after this visit. An endoscopy revealed pyloric stenosis, which did not improve with balloon dilatation. We performed a gastro-jejunal and cecal-transverse colonic bypass surgery. Diffuse wall thickening of the antrum was verified during the surgery, and a biopsy sample was collected. The diagnosis of gastric metastasis from breast was confirmed since it showed the same immunohistochemistry pattern as the prior breast lesion. Pyloric stenosis has still been confirmed with an endoscopy, she has been alive with satisfactory oral food intake for >10 years. [ABSTRACT FROM AUTHOR]

S100 proteins convey important roles in innate immune responses to infection and regenerative processes. However, their role in inflammatory or regenerative processes of the human dental pulp is poorly elucidated. The aim of the present study was to detect, localize, and compare the occurrence of 8 S100 proteins in normal, symptomatic, and asymptomatic irreversibly inflamed dental pulp specimens. Human dental pulp specimens from 45 individuals were clinically assigned to 3 groups of pulpal diagnosis: normal pulp (NP, n = 17), asymptomatic irreversible pulpitis (AIP, n = 13), and symptomatic irreversible pulpitis (SIP, n = 15). The specimens were prepared and immunohistochemically stained for proteins S100A1, -A2, -A3, -A4, -A6, -A7, -A8, and -A9. Staining was classified using semiquantitative analysis and a 4-degree staining score (ie, no, decent, medium, and intense staining) at 4 different anatomic or functional regions (ie, the odontoblast layer [OL], pulpal stroma [PS], border area of calcifications [BAC], and vessel walls). The distribution of staining degrees between the 3 diagnostic groups was calculated using the Fisher exact text (P ≤.05) at the 4 regions. Significant differences in staining were observed mainly in the OL and PS and at the BAC. The most significant differences were detected in the PS and when comparing NP with 1 of the 2 irreversibly inflamed pulpal tissues (AIP or SIP). The inflamed tissues were then invariably stained more intensely than their normal counterparts at this location (S100A1, -A2, -A3, -A4, -A8, and -A9). In the OL, NP tissue was significantly stronger stained for S100A1, -A6, -A8, and -A9 compared with SIP and for S100A9 when compared with AIP. Differences between AIP and SIP in direct comparison were rare and were found only for 1 protein (S100A2) at the BAC. Also, at the vessel walls, only 1 statistical difference in staining was observed (SIP was stronger stained than NP for protein S100A3). The occurrence of proteins S100A1, -A2, -A3, -A4, -A6, -A8, and -A9 is significantly altered in irreversibly inflamed compared with normal dental pulp tissue at different anatomic localizations. Some members of S100 proteins obviously participate in focal calcification processes and pulp stone formation of the dental pulp. [ABSTRACT FROM AUTHOR]

Introduction Squamous cell carcinoma arising from a suprapubic cystostomy tract is a rare complication of an indwelling catheter and is caused by long‐term inflammation and mechanical irritation. Prognosis is relatively poor. Biomarkers in the cancer pathway have not been investigated. Case presentation A 61‐year‐old woman with a 34‐year history of suprapubic catheter placement presented with a rapidly growing elevated lesion around the cystostomy site. Tumor biopsy confirmed squamous cell carcinoma. Local excision with partial cystectomy was performed. Multiple metastases were identified 5 months later. The patient died 14 months after the initial treatment. Immunohistochemical analysis of the resected specimen revealed alterations in vascular endothelial growth factor, epidermal growth factor receptor, cyclooxygenase‐2, and Ki‐67. Conclusion We encountered a case of squamous cell carcinoma arising from a suprapubic cystostomy tract. Immunohistochemical analysis revealed activation of multiple carcinogenic pathways in cancer cells, including those for angiogenesis, signal transduction by epidermal growth factor receptor, inflammation, and cell proliferation.

Abstract Primary extranodal diffuse large B‐cell lymphoma (PE‐DLBCL) is a heterogeneous subgroup of DLBCL. We investigated the prevalence and prognostic value of surface expression of PD‐L1, PD1, and CD30, copy number of 9p24.1 (PD‐L1 region), and mutations in MYD88, CD79B, CARD11, and BTK in a cohort of 116 patients, localized in the mediastinum (PMBL, n = 12), ear, nose and throat (ENT, n = 28), central nervous system (n = 29), testis (n = 7), breast (n = 4), stomach (n = 10), bone (n = 8), spleen (n = 2), and skin (n = 16). PD‐L1 expression is most frequent in PMBL (92%), followed by lymphomas originating in the stomach (57%), ENT (23%), and skin (18%). PD1 was expressed at low levels in less than 13% of PE‐DLBCL, while CD30 expression was found in 58% of PMBL. Mutation analysis revealed an unexpectedly high frequency of MYD88 and CD79B mutations in ENT lymphomas (46% and 50%, respectively). CARD11 mutations are rare but more frequently found in gastric lymphomas (30%), suggesting BTK resistance. Thirty‐four of 113 (30%) of the lymphomas harbored both MYD88 and CD79B mutations. Lower overall and progression‐free survival rates were found for cases with MYD88, CD79B, and BTK mutations. These data confirm the biologic singularity of PE‐DLBCLs and provide some suggestions for targeted therapies.

Glioblastoma is the most common malignant brain tumor in adults. Standard treatment includes tumor resection, radio-chemotherapy and adjuvant chemotherapy with temozolomide (TMZ). TMZ methylates DNA, whereas O6-methylguanine DNA methyltransferase (MGMT) counteracts TMZ effects by removing the intended proteasomal degradation signal. Non-functional MGMT mediates the mismatch repair (MMR) system, leading to apoptosis after futile repair attempts. This study investigated the associations between MGMT promoter methylation, MGMT and MMR protein expression, and their effect on overall survival (OS) and progression-free survival (PFS) in patients with glioblastoma. MGMT promoter methylation was assessed in 42 treatment-naïve patients with glioblastoma WHO grade IV by pyrosequencing. MGMT and MMR protein expression was analyzed using immunohistochemistry. MGMT promoter methylation was present in 52%, whereas patients <70 years of age revealed a significantly longer OS using a log-rank test and a significance threshold of p ≤ 0.05. MGMT protein expression and methylation status showed no correlation. MMR protein expression was present in all patients independent of MGMT status and did not influence OS and PFS. Overall, MGMT promoter methylation implicates an improved OS in patients with glioblastoma aged <70 years. In the elderly, the extent of surgery has an impact on OS rather than the MGMT promoter methylation or protein expression. [ABSTRACT FROM AUTHOR]

Introduction: Glucocorticosteroid therapy is the basic therapeutic option for rheumatoid arthritis; however, long-term treatment with low and medium doses is associated with the development of negative side effects. As reported, oxidative stress is crucial in the pathogenesis of rheumatoid arthritis. Therefore, pharmacotherapy that combines application of methylprednisolone pulse therapy and an aqueous molecular hydrogen solution seems to be a reasonable treatment option in these cases. Materials and Methods: The study of the effectiveness of combined pharmacotherapy for rheumatoid arthritis was carried out on mature male rats. Rheumatoid arthritis was simulated by introducing bovine type II collagen into the right knee joint. Animals of the 1st - control - group received placebo; animals of the 2nd group received molecular hydrogen-enriched water intragastrically; animals of the 3rd group received methylprednisolone solution intravenously by catheterization of the tail vein, and animals of the 4th group received water enriched with molecular hydrogen intragastrically and methylprednisolone solution intravenously. After withdrawal of animals from the experiment, microsections of their joint tissues were analysed histologically and biomarkers of the joint inflammation were detected immunohistochemically. Results and Discussion: Morphological analysis of microsections taken from animals of the 4th group evidenced effectiveness of the combined therapy based on quantitative estimation of the inflammatory biomarker expression. Dynamic polarization of M1/M2 macrophage was manifested with high quality in animals of this group. Conclusion: The search for new therapeutic options for rheumatoid arthritis is expanded due to major antioxidant substances that can reduce duration of treatment, while ensuring positive dynamics of the course of the disease. [ABSTRACT FROM AUTHOR]

Background and objectives: In patients with colorectal cancer (CRC), heterogeneous expression of Mismatch repair (MMR) proteins can manifest itself in several different forms and is not such a rare phenomenon. Therefore, it is very important to recognize the nuclear expression of MMR proteins of different MMR status in order to avoid false positive or false negative results. The aim of this study was to determine the frequency and distribution of heterogeneous expression of MMR proteins in patients with stages II and III of the disease as well as its association with clinical, demographic and pathological characteristics of CRC in relation to proficient and deficient expression of MMR proteins. Material and Methods: The study included 104 cases of colorectal cancer obtained from surgical colectomy material in stages II and III of the disease. Results: From a total of 104 patients with colorectal cancer, immunohistochemical analysis of the expression of all four MMR proteins showed that heterogeneous expression of MMR proteins (as well as deficient immunoreactivity of tumor cells) was present in 12 cases, while proficient expression of MMR proteins was detected in 80 tumors. Conclusions: Our study showed that the only independent predictors of the loss of MMR protein expression were younger patient age and right-sided anatomical location of the tumor. The study also established the existence of heterogeneous expression of MMR proteins in a non-negligible percentage of CRCs (11.5%), where heterogeneous nuclear expression of MMR proteins was described in several different forms. [ABSTRACT FROM AUTHOR]

The identification of markers for early diagnosis, prognosis, and improvement of therapeutic options represents an unmet clinical need to increase survival in Non-Small Cell Lung Cancer (NSCLC), a neoplasm still characterized by very high incidence and mortality. Here, we investigated whether proline dehydrogenase (PRODH), a mitochondrial flavoenzyme catalyzing the key step in proline degradation, played a role in NSCLC tumorigenesis. PRODH expression was investigated by immunohistochemistry; digital PCR, quantitative PCR, immunoblotting, measurement of reactive oxygen species (ROS), and functional cellular assays were carried out. PRODH expression was found in the majority of lung adenocarcinomas (ADCs). Patients with PRODH-positive tumors had better cancer-free specific and overall survival compared to those with negative tumors. Ectopic modulation of PRODH expression in NCI-H1299 and the other tested lung ADC cell lines decreased cell survival. Moreover, cell proliferation curves showed delayed growth in NCI-H1299, Calu-6 and A549 cell lines when PRODH-expressing clones were compared to control clones. The 3D growth in soft agar was also impaired in the presence of PRODH. PRODH increased reactive oxygen species production and induced cellular senescence in the NCI-H1299 cell line. This study supports a role of PRODH in decreasing survival and growth of lung ADC cells by inducing cellular senescence.

Trophoblast cell surface antigen 2 (Trop-2, encoded by TACSTD2) is the target protein of sacituzumab govitecan, a novel antibody-drug conjugate for locally advanced or metastatic urothelial carcinoma. However, the expression status of Trop-2 in upper tract urothelial carcinoma (UTUC) remains unclear. We performed immunohistochemical analysis of 99 UTUC samples to evaluate the expression status of Trop-2 in patients with UTUC and analyze its association with clinical outcomes. Trop-2 was positive in 94 of the 99 UTUC samples, and high Trop-2 expression was associated with favorable progression-free survival (PFS) and cancer-specific survival (p = 0.0011, 0.0046). Multivariate analysis identified high Trop-2 expression as an independent predictor of favorable PFS (all cases, p = 0.045; high-risk group (pT3≤ or presence of lymphovascular invasion or lymph node metastasis), p = 0.014). Gene expression analysis using RNA sequencing data from 72 UTUC samples demonstrated the association between high TACSTD2 expression and favorable PFS (all cases, p = 0.069; high-risk group, p = 0.029). In conclusion, we demonstrated that Trop-2 is widely expressed in UTUC. Although high Trop-2 expression was a favorable prognostic factor in UTUC, its widespread expression suggests that sacituzumab govitecan may be effective for a wide range of UTUC.

The objective was to develop an automated algorithm for the estimation of a protein (Ki-67) index based on cell nuclei area ratio of gastric epithelial tissue cells; for this purpose, digital histopathology images were used. An expert manually annotated each region of interest of the images. A proportion of Ki-67 positive and negative cells within that region was used to obtain the color distribution of the corresponding pixels. The histogram of each color distribution was modeled as a Gaussian and, later, thresholded for segmentation and classification. Finally, the Ki-67 index was estimated as the ratio between the segmented positive area of the nuclei divided by the total area of the positive and negative nuclei. The automated method has a strong correlation of 0.725 and a root mean square error of 0.293 when compared to the manual method, which gives certainty that the automated method can be used to analyze the proliferation rate. Furthermore, compared to manual classification, the presented method automatically classifies every image in the same Ki-67 category: low, intermediate, and high. Despite the small sample size, the utility of the presented method was demonstrated. However, the low number of scored images did not allow for thoroughly sampling the ranges of pixel values and intensities observed by pathologists, which will be addressed in future work.

A comprehensive molecular classification was published in 2014 within The Cancer Genome Atlas (TCGA) to guide clinical approaches and treatment strategies. This study aimed to investigate the clinicopathological and prognostic importance of the classification using immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH) to identify potential surrogate markers of molecular changes in gastroesophageal junction (GEJ) adenocarcinomas. A total of 52 GEJ adenocarcinomas were divided into five groups using IHC with MLH-1, E-cadherin, p53 and CISH with EBER: 1) microsatellite unstable (MSI: negative with MLH-1), 2) genomically stable tumors (GS: positive with p53), 3) chromosomally unstable tumors (CUN: negative with e-cadherin), 4) EBV+ tumors (EBV+: positive with EBER) and 5) unclassifiable (G-NOS: MLH-1 and e-cadherin positive with p53 and negative with EBER). The largest group consisted of 24 (46.2%) cases of CUN tumors. This group was followed by groups of GS with 14 (26.9%) cases, MSI with 7 (13.5%) cases, and EBV + with 3 (5.8%) cases, respectively. Although this classification was not associated with pathological features, it was found to be closely related to prognosis (p = 0.029). Patients with EBV+ tumors had the longest overall survival, followed by the G-NOS, MSI, CUN, GS groups.

The expression level of retinol‐binding protein 4 (RBP4) protein is closely related to liver damage and plays an important role in the diagnosis and prognosis of cancer. However, the preparation of anti‐RBP4 mAb or exploration on the application of anti‐RBP4 mAb has not been reported thus far. In the present study, we constructed a pET30a‐RBP4 recombinant vector, used E. coli BL21 (DE3) as the vector to express the RBP4 recombinant protein and prepared anti‐RBP4 mAb using hybridoma technology. We performed immunohistochemical analysis on hepatocellular carcinoma (HCC) and adjacent tissues by using this anti‐RBP4 mAb. In addition to the high‐purity RBP4 recombinant protein, we successfully developed the anti‐RBP4 mAb with high affinity and specificity; it binds to natural RBP4 protein and is suitable for immunohistochemical analysis.

Carcinoma of the extrahepatic biliary tract accounts for

Background: Trophoblast cell-surface antigen 2 (TROP2) is expressed on the surface of trophoblast cells and many malignant tumor cells. However, data on TROP2 expression in advanced lung cancer are insufficient, and its changes have not been fully evaluated. Methods: We assessed the prevalence and changes in TROP2 expression in patients with lung cancer who received anti-cancer treatments using immunohistochemical (IHC) analysis with an anti-TROP2 antibody (clone: SP295). IHC scores were graded from 0 to 3; grade ≥ 2 was considered positive for TROP2 expression. We defined a difference in IHC score, before and after anti-cancer treatments, as the change in TROP2 expression. Results: Before anti-cancer treatment, TROP2 expression was observed in 89% (143/160) of the patients and was significantly more common in adenocarcinoma and squamous cell carcinoma than in neuroendocrine carcinoma (P < 0.001). After anti-cancer treatment, TROP2 expression was observed in 87% (139/160) of the patients. The distribution of TROP2 expression in post-treatment samples was analogous to that in pre-treatment samples when compared using the Wilcoxon signed-rank test (P = 0.509). However, an increase in TROP2 expression was seen in 19 (12%), and a decrease in 20 (13%) patients. Patients treated with targeted therapy showed significantly higher changes in TROP2 expression (P = 0.019) and thoracic radiotherapy was more likely to increase TROP2 expression than chemotherapy alone. Conclusion: Although some anti-cancer treatments might alter the TROP2 expression, TROP2 was expressed in most lung cancer specimens before and after anti-cancer treatments. These results support the development of TROP2-directed therapy against advanced lung cancer in various treatment lines. [ABSTRACT FROM AUTHOR]

Myocardial injury in patients with SARS-CoV-2 infection may be attributed to the presence of the virus at the cellular level, however, it may also be secondary to other diseases, playing an essential role in the evolution of the disease. We evaluated 16 patients who died because of SARS-CoV-2 infection and analyzed the group from both clinical and pathological points of view. All autopsies were conducted in the Sibiu County morgue, taking into consideration all the national protocols for COVID-19 patients. Of the 16 autopsies we performed, two were complete, including an extensive examination of the cranial cavity. In our study, the cardiac injury was primarily cumulative. Chronic cardiac injuries included fatty infiltration of the myocardium in five cases, fibrosis in 11 cases, and coronary atherosclerosis in two cases. Among the cases with evidence of acute cardiovascular injuries, inflammatory lymphocytic infiltrate was observed in nine cases, subepicardial or visceral pericardial neutrophil-rich vascular congestion in five cases, and venous thrombosis in three cases. Acute ischemia or myocytic distress was identified by vacuolar degeneration in four cases; areas of undulated and/or fragmented myocardial fibers, with eosinophilia and nuclear pyknosis with or without enucleation of the myocytes in nine cases; and in one case, we observed a large area of myocardial necrosis. Immunohistochemical criteria confirmed the presence of the SARS-CoV-2 antigen at the level of the myocardium in only two cases. Comorbidities existing prior to SARS-CoV-2 infection associated with systemic and local inflammatory, thrombotic, hypoxic, or immunological phenomena influence the development of cardiac lesions, leading to death. [ABSTRACT FROM AUTHOR]

Purpose: In clinical use of low-level laser therapy for bone regeneration (LLLT), application protocol (dose, duration, and repetitions) has not been established. This study aimed to depict a reliable dosage of LLLT by evaluating the efficacy of different dosing of LLLT (diode) on the healing of rabbit cranial defects. Methods: Critical size defects were prepared in calvarias of 26 New Zealand White Rabbits in such each animal containing both test and control groups. Test groups were irradiated with 4 Joule/cm2 (j/cm2), 6 j/cm2, and 8 j/cm2. The rabbits were subjected to six times of laser treatments in 10 days. At the end of the second week, 5 rabbits were sacrificed for histopathological and immunohistochemical analyses. At the 4th and 8th weeks, 20 rabbits (10 each) were sacrificed for micro-CT and histopathological analyses. Results: Micro-CT evaluation revealed improved new bone formation in all test groups compared to the control group. 6 j/cm2 group demonstrated the highest bone formation. The highest bone morphogenic protein -2 levels were found in the 4 j/cm2 group. Osteocalcin expression was significantly higher in 4 j/cm2 group. Conclusions: Our findings indicate that LLLT have a positive effect on new bone formation. The high efficacy of doses of 4 j/cm2 and 6 j/cm2 is promising to promote early bone healing. [ABSTRACT FROM AUTHOR]

Trophoblast cell surface antigen 2 (Trop-2, encoded by TACSTD2) is the target protein of sacituzumab govitecan, a novel antibody-drug conjugate for locally advanced or metastatic urothelial carcinoma. However, the expression status of Trop-2 in upper tract urothelial carcinoma (UTUC) remains unclear. We performed immunohistochemical analysis of 99 UTUC samples to evaluate the expression status of Trop-2 in patients with UTUC and analyze its association with clinical outcomes. Trop-2 was positive in 94 of the 99 UTUC samples, and high Trop-2 expression was associated with favorable progression-free survival (PFS) and cancer-specific survival (p = 0.0011, 0.0046). Multivariate analysis identified high Trop-2 expression as an independent predictor of favorable PFS (all cases, p = 0.045; high-risk group (pT3≤ or presence of lymphovascular invasion or lymph node metastasis), p = 0.014). Gene expression analysis using RNA sequencing data from 72 UTUC samples demonstrated the association between high TACSTD2 expression and favorable PFS (all cases, p = 0.069; high-risk group, p = 0.029). In conclusion, we demonstrated that Trop-2 is widely expressed in UTUC. Although high Trop-2 expression was a favorable prognostic factor in UTUC, its widespread expression suggests that sacituzumab govitecan may be effective for a wide range of UTUC. [ABSTRACT FROM AUTHOR]

Abstract Microanatomy of the vast majority of human organs at birth is characterized by marked differences as compared to adult organs, regarding their architecture and the cell types detectable at histology. In preterm neonates, these differences are even more evident, due to the lower level of organ maturation and to ongoing cell differentiation. One of the most remarkable finding in preterm tissues is the presence of huge amounts of stem/progenitor cells in multiple organs, including kidney, brain, heart, adrenals, and lungs. In other organs, such as liver, the completely different burden of cell types in preterm infants is mainly related to the different function of the liver during gestation, mainly focused on hematopoiesis, a function that is taken by bone marrow after birth. Our preliminary studies showed that the antigens expressed by stem/progenitors differ significantly from one organ to the next. Moreover, within each developing human tissue, reactivity for different stem cell markers also changes during gestation, according with the multiple differentiation steps encountered by each progenitor during development. A better knowledge of stem/progenitor cells of preterms will allow neonatologists to boost preterm organ maturation, favoring the differentiation of the multiple cells types that characterize each organ in at term neonates.

Technological advances have allowed the discovery of 6 subtypes of membranous nephropathy based on target antigens: M-type phospholipase A2 receptor (PLA2R), thrombospondin type 1 domain-containing 7A (THSD7A), neural epidermal growth factor-like 1 protein, semaphorin 3B, exostosin 1 (EXT1), and EXT2. EXT1/EXT2 are thought to be associated with secondary (autoimmune) membranous nephropathy. Although it has been reported that PLA2R- and THSD7A-associated membranous nephropathy have rarely been detected concomitantly, there have been no previous reports demonstrating PLA2R- or THSD7A-associated membranous nephropathy with enhanced glomerular staining of EXT1/EXT2. We describe 2 cases of primary membranous nephropathy with enhanced glomerular staining of EXT1/EXT2. Patient 1 was diagnosed with PLA2R-associated primary membranous nephropathy, and patient 2 was diagnosed with THSD7A-associated primary membranous nephropathy. Both patients achieved clinical remission in response to immunosuppressive therapy. Neither patient demonstrated signs of autoimmune diseases, and antinuclear antibodies were absent in their sera. Based on these 2 cases, enhanced staining of EXT1/EXT2 in glomeruli, although rare, can be detected in primary membranous nephropathy without autoimmune diseases.

Background. Currently, there are several forms of breast cancer (BC): nodular, diffuse, edematous-infiltrative, mastitis-like form, as well as Paget’s cancer. A special form of BC is the hidden or occult form. Taking into account the analysis of methods for the diagnosis and treatment of occult BC, it is important that this form of the disease is detected at a later stage. It includes axillary lymphadenopathy and histologically appears to be a metastatic lesion of the lymph nodes from undetectable invasive BC.Materials and methods. We conducted a thorough analysis of domestic and foreign literature and described two clinical examples with authentic documentation of the results of the examination and treatment. The article describes in detail the schemes of drug treatment and descriptions of the results of morphological examination of the surgical material.Results. Given the complexity of the diagnostic stage, the clinical form of the course of diseases, the described clinical examples can be attributed specifically to the occult form of BC. An important and determining factor in treatment is the diagnostic stage using all possible methods, including positron emission tomography–computed tomography, breast magnetic resonance imaging and trepan-biopsy of regional lymph nodes, followed by the determination of the molecular biological subtype of the tumor.Conclusions. In this article, the presented data confirm the general statement about the uniqueness of this disease, which requires a more detailed and multidisciplinary approach at the stage of diagnosis and determination of treatment tactics.

Primary mediastinal sarcomas are extremely rare. Additionally, mediastinal leiomyosarcomas account for approximately 9% of mediastinal sarcoma cases. Until date, only few cases of anterior mediastinal leiomyosarcomas have been reported. Herein, we report a case of an 85-year-old female with an anterior mediastinal mass of 15 mm. Histological examination revealed spindle tumor cells showing a fascicular growth pattern. Immunohistochemically, the tumor cells were focal positive for desmin, calponin, and α-smooth muscle actin. The pathological diagnosis was leiomyosarcoma. In conclusion, we encountered a case of a very rare leiomyosarcoma that occurred in the anterior mediastinum, and our report may contribute to the understanding of this disease.

Glioblastoma is the most common malignant brain tumor in adults. Standard treatment includes tumor resection, radio-chemotherapy and adjuvant chemotherapy with temozolomide (TMZ). TMZ methylates DNA, whereas O6-methylguanine DNA methyltransferase (MGMT) counteracts TMZ effects by removing the intended proteasomal degradation signal. Non-functional MGMT mediates the mismatch repair (MMR) system, leading to apoptosis after futile repair attempts. This study investigated the associations between MGMT promoter methylation, MGMT and MMR protein expression, and their effect on overall survival (OS) and progression-free survival (PFS) in patients with glioblastoma. MGMT promoter methylation was assessed in 42 treatment-naïve patients with glioblastoma WHO grade IV by pyrosequencing. MGMT and MMR protein expression was analyzed using immunohistochemistry. MGMT promoter methylation was present in 52%, whereas patients p ≤ 0.05. MGMT protein expression and methylation status showed no correlation. MMR protein expression was present in all patients independent of MGMT status and did not influence OS and PFS. Overall, MGMT promoter methylation implicates an improved OS in patients with glioblastoma aged MGMT promoter methylation or protein expression.

Background and objectives: In patients with colorectal cancer (CRC), heterogeneous expression of Mismatch repair (MMR) proteins can manifest itself in several different forms and is not such a rare phenomenon. Therefore, it is very important to recognize the nuclear expression of MMR proteins of different MMR status in order to avoid false positive or false negative results. The aim of this study was to determine the frequency and distribution of heterogeneous expression of MMR proteins in patients with stages II and III of the disease as well as its association with clinical, demographic and pathological characteristics of CRC in relation to proficient and deficient expression of MMR proteins. Material and Methods: The study included 104 cases of colorectal cancer obtained from surgical colectomy material in stages II and III of the disease. Results: From a total of 104 patients with colorectal cancer, immunohistochemical analysis of the expression of all four MMR proteins showed that heterogeneous expression of MMR proteins (as well as deficient immunoreactivity of tumor cells) was present in 12 cases, while proficient expression of MMR proteins was detected in 80 tumors. Conclusions: Our study showed that the only independent predictors of the loss of MMR protein expression were younger patient age and right-sided anatomical location of the tumor. The study also established the existence of heterogeneous expression of MMR proteins in a non-negligible percentage of CRCs (11.5%), where heterogeneous nuclear expression of MMR proteins was described in several different forms.

Introduction: The specific pathogenesis of ankylosing spondylitis (AS) remains unclear, and our study aimed to investigate the possible pathogenesis of AS. Materials and Methods: Two datasets were downloaded from the GEO database to perform differentially expressed gene analysis, GO enrichment analysis, KEGG pathway analysis, DO enrichment analysis, GSEA analysis of differentially expressed genes, and construction of diagnostic genes using SVM and WGCNA along with Hypoxia-related genes. Also, drug sensitivity analysis was performed on diagnostic genes. To identify the differentially expressed immune genes in the AS and control groups, we analyzed the composition of immune cells between them. Then, we examined differentially expressed genes in three AS interspinous ligament specimens and three Degenerative lumbar spine specimens using high-throughput sequencing while the immune cells were examined using the neutrophil count data from routine blood tests of 1770 HLA-B27-positive samples and 7939 HLA-B27-negative samples. To assess the relationship between ANXA3 and SORL1 and disease activity, we took the neutrophil counts of the first 50 patients with above-average BASDAI scores and the last 50 patients with below-average BASDAI scores for statistical analysis. We used immunohistochemistry to verify the expression of ANXA3 and SORL1 in AS and in controls. Results: ANXA3 and SORL1 were identified as new diagnostic genes for AS. These two genes showed a significant differential expression between AS and controls, along with showing a significant positive correlation with the neutrophil count. The results of high-throughput sequencing verified that these two gene deletions were indeed differentially expressed in AS versus controls. Data from a total of 9707 routine blood tests showed that the neutrophil count was significantly higher in AS patients than in controls (p < 0.001). Patients with AS with a high BASDAI score had a much higher neutrophil count than those with a low score, and the difference was statistically significant (p < 0.001). The results of immunohistochemistry showed that the expression of ANXA3 and SORL1 in AS was significantly higher than that in the control group. Conclusion: Upregulated of ANXA3, SORL1, and neutrophils may be a key factor in the progression of Ankylosing spondylitis. [ABSTRACT FROM AUTHOR]

The expression level of retinol‐binding protein 4 (RBP4) protein is closely related to liver damage and plays an important role in the diagnosis and prognosis of cancer. However, the preparation of anti‐RBP4 mAb or exploration on the application of anti‐RBP4 mAb has not been reported thus far. In the present study, we constructed a pET30a‐RBP4 recombinant vector, used E. coli BL21 (DE3) as the vector to express the RBP4 recombinant protein and prepared anti‐RBP4 mAb using hybridoma technology. We performed immunohistochemical analysis on hepatocellular carcinoma (HCC) and adjacent tissues by using this anti‐RBP4 mAb. In addition to the high‐purity RBP4 recombinant protein, we successfully developed the anti‐RBP4 mAb with high affinity and specificity; it binds to natural RBP4 protein and is suitable for immunohistochemical analysis. [ABSTRACT FROM AUTHOR]

The aim of this study was to quantify the distribution of microvessels and mast cells in all three parts of the trigeminal ganglion (TG). Statistical analyses were applied to investigate possible micromorphological regional differences in their density. Five serially sectioned human TGs were prepared for CD34 and mast cell tryptase immunostaining. The following quantifications were performed in microscopic fields of three parts of the TG: microvessel density (MVD), mast cell density (MCD) and ganglionic cell count. The density of CD34-positive microvessels was not significantly different in any of the three observed parts of the TG. The distribution of neurons showed no significant statistical difference in three parts of the TG. There was no difference in the density of tryptase-positive mast cells within the TG, but there was an abundant presence of mast cells in the periganglionic dural and subdural tissues, a finding hitherto not reported. We can say that there is a homogenous vascular pattern within the TG which excludes local predominance in pathogenesis of trigeminal neuralgia. Second, and more important, the finding of peri-trigeminal mast cells indicates their important role in migraine pain and confirms their degranulation as the main therapeutic goal for this condition.

IntroductionThe specific pathogenesis of ankylosing spondylitis (AS) remains unclear, and our study aimed to investigate the possible pathogenesis of AS.Materials and MethodsTwo datasets were downloaded from the GEO database to perform differentially expressed gene analysis, GO enrichment analysis, KEGG pathway analysis, DO enrichment analysis, GSEA analysis of differentially expressed genes, and construction of diagnostic genes using SVM and WGCNA along with Hypoxia-related genes. Also, drug sensitivity analysis was performed on diagnostic genes. To identify the differentially expressed immune genes in the AS and control groups, we analyzed the composition of immune cells between them. Then, we examined differentially expressed genes in three AS interspinous ligament specimens and three Degenerative lumbar spine specimens using high-throughput sequencing while the immune cells were examined using the neutrophil count data from routine blood tests of 1770 HLA-B27-positive samples and 7939 HLA-B27-negative samples. To assess the relationship between ANXA3 and SORL1 and disease activity, we took the neutrophil counts of the first 50 patients with above-average BASDAI scores and the last 50 patients with below-average BASDAI scores for statistical analysis. We used immunohistochemistry to verify the expression of ANXA3 and SORL1 in AS and in controls.ResultsANXA3 and SORL1 were identified as new diagnostic genes for AS. These two genes showed a significant differential expression between AS and controls, along with showing a significant positive correlation with the neutrophil count. The results of high-throughput sequencing verified that these two gene deletions were indeed differentially expressed in AS versus controls. Data from a total of 9707 routine blood tests showed that the neutrophil count was significantly higher in AS patients than in controls (p < 0.001). Patients with AS with a high BASDAI score had a much higher neutrophil count than those with a low score, and the difference was statistically significant (p < 0.001). The results of immunohistochemistry showed that the expression of ANXA3 and SORL1 in AS was significantly higher than that in the control group.ConclusionUpregulated of ANXA3, SORL1, and neutrophils may be a key factor in the progression of Ankylosing spondylitis.

Objective of the study: to study peculiarities of the inflammatory reactions development in placenta according to results of a general histological and immunohistochemical study of the prevalence and intensity of reactions caused by CD45 and tumor necrosis factor α (TNF-α) during childbirth at different gestational periods. Materials and methods. Morphological and immunohistochemical studies of various placental structures and CD45 and TNF-α expression level in 142 women with preterm delivery at different gestational periods were performed: group I – at 23–29 weeks, group II – at 30–34 weeks; group III – at 35–36 weeks + 6 days, group IV (control) – 40 women at 38–40 weeks. Results. Inflammatory infiltration and the most expressive CD45 expression prevailed in the decidual membrane, TNF-α expression was predominantly negative in group I. In group II, inflammatory changes were evidenced by collagenization and stromal fibrosis, a decrease in fetal vessels, fibroblast proliferation, an increase in intervillous fibrinoid, and CD45 spread was expressed in decreasing order of intensity in decidual membrane, in stroma of intermediate villi and in stem villi. In group III, foci of inflammatory infiltration spread from the decidual membrane to the intervillous space and villi stroma, vessels plethora of all types of villi, dystrophic changes, and fibrinoid volume increased. CD45 expression was more common and expressive than in the groups I and II, and TNF-α expression was weakly positive only in the decidual membrane. The control group determined the widespread expressive CD45 expression in all placental structures and positive moderate TNF-α expression. Conclusions. Preterm delivery and delivery on term occur when there is an activation of inflammatory reactions in placenta. Inflammation in the placental structures is a necessary condition for development of the mother-fetal interphase, however, the features of local immune response prove the need to determine genetic mechanisms of the preterm labor.

Abstract Background Clear cell tumors of the lung (CCTLs) are rare and mostly benign pulmonary neoplasms arising from perivascular epithelioid cells. Only approximately 100 cases have been reported, and half of them were in China. Limited details about CCTLs often cause diagnostic or therapeutic problems. Case presentation We describe a case of a 28-year-old woman with multiple gradually replicating and enlarging nodules in the left lower lobe. The patient underwent fine-needle aspiration biopsy and was diagnosed with CCTL. A left lower lobectomy and mediastinal lymph node dissection were performed. The gradual changes in size (1.4 cm to 2.8 cm) and quantity (10 to 49) of the CCTLs in this case were the biggest differences from previously reported cases. Conclusions CCTLs are very uncommon and mostly benign PEComatous tumors with no specific morphologic features. We present a case of CCTL with multiplicity and rapid growth, which may indicate its aggressive nature. The accumulation of similar cases will help clarify the exact nature and improve our understanding of the disease.

Nuclear protein in testis (NUT) carcinoma is a rare, aggressive carcinoma that is a diagnostic challenge for pathologists. Here, we report a case of NUT carcinoma in a 63‐year‐old woman with uncommon immunohistochemical results. The initial bronchoscopic biopsy revealed a poorly differentiated carcinoma with p63 immunohistochemical stain positivity. However, the cytomorphological features of the pleural fluid were unusual. Immunohistochemical staining of the pleural fluid revealed diffuse positivity for vimentin and focal positivity for cytokeratin and neuroendocrine markers. Because of chemoresistance, other malignancies, including sarcomatoid carcinoma, combined small cell carcinoma, and an unusual form of NUT carcinoma, were considered as differential diagnoses. The diagnosis of NUT carcinoma was confirmed using NUT‐specific antibodies and fluorescence in situ hybridization. The current case was a diagnostic challenge because of the poorly differentiated cytomorphology and uncommon immunohistochemical results. Pathologists and clinicians should consider NUT carcinoma in the differential diagnosis, as this malignancy has a dismal prognosis and needs to be diagnosed accurately for the most effective treatment. Key points Metastatic NUT carcinoma can show diffuse vimentin positivity and focal neuroendocrine marker positivity. NUT carcinoma can be misdiagnosed as basaloid squamous cell carcinoma in routine diagnosis, especially in older‐aged patients. This study was a diagnostic challenge because of the poorly differentiated cytomorphology and uncommon immunohistochemical results for NUT carcinoma. Pathologists should differentially diagnose NUT carcinoma when rare cytohistological features are observed at any age.

A comprehensive molecular classification was published in 2014 within The Cancer Genome Atlas (TCGA) to guide clinical approaches and treatment strategies. This study aimed to investigate the clinicopathological and prognostic importance of the classification using immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH) to identify potential surrogate markers of molecular changes in gastroesophageal junction (GEJ) adenocarcinomas. A total of 52 GEJ adenocarcinomas were divided into five groups using IHC with MLH-1, E-cadherin, p53 and CISH with EBER: 1) microsatellite unstable (MSI: negative with MLH-1), 2) genomically stable tumors (GS: positive with p53), 3) chromosomally unstable tumors (CUN: negative with e-cadherin), 4) EBV+ tumors (EBV+: positive with EBER) and 5) unclassifiable (G-NOS: MLH-1 and e-cadherin positive with p53 and negative with EBER). The largest group consisted of 24 (46.2%) cases of CUN tumors. This group was followed by groups of GS with 14 (26.9%) cases, MSI with 7 (13.5%) cases, and EBV + with 3 (5.8%) cases, respectively. Although this classification was not associated with pathological features, it was found to be closely related to prognosis (p = 0.029). Patients with EBV+ tumors had the longest overall survival, followed by the G-NOS, MSI, CUN, GS groups. [ABSTRACT FROM AUTHOR]