Staining Intensity Research Articles

TLR4 Downregulation Identifies High-Risk HPV Infection and Integration in H-SIL and Squamous Cell Carcinomas of the Uterine Cervix.

Growing scientific evidence suggests a link between the expression of toll-like receptor 4 (TLR4) and cervical cancer carcinogenesis. Specifically, a close relation between TLR4 expression and FIGO stage, lymph node metastases, and tumor size has been reported in cervical cancer. In the present study, we aimed to evaluate the relationship between TLR4 expression levels and human papillomavirus (HPV) infection and/or high-risk (hr) HPV integration status in patients with a histological diagnosis of high-grade squamous intraepithelial lesion (H-SIL), and squamous cell carcinoma (SCC) of the uterine cervix. Sixty biopsies of cervical neoplasia, comprising H-SIL (n = 20) and SCC (n = 40), were evaluated for TLR4 expression by immunohistochemistry. All samples were positive for high-risk HPV as confirmed by in situ hybridization (ISH) and broad-spectrum PCR followed by Sanger sequencing analysis. The intensity of TLR4 staining was higher in tissues negative for intraepithelial lesion or malignancy (NILM) than in H-SIL, and further reduced in SCC. Moreover, statistically significant differences have been observed in the percentage of TLR4 expression between NILM and H-SIL and between H-SIL and SCC, with higher percentages of expression in H-SIL than in SCC. Our results showed a significant downregulation of TLR4 in HPV-related H-SIL and SCC, compared to NILM. These data support the hypothesis that TLR4 expression is suppressed in HPV-driven oncogenesis.

Predictive value of methylene blue combined with indocyanine green in sentinel lymph node metastasis in breast cancer: a prospective pilot cohort study.

The status of sentinel lymph nodes is crucial for prognosis and treatment decisions in breast cancer patients. This study aimed to evaluate the predictive value of combined methylene blue and indocyanine green for sentinel lymph node metastasis in breast cancer. This prospective cohort study enrolled 90 clinically node-negative breast cancer patients. Methylene blue and indocyanine green were injected locally before surgery. Sentinel lymph nodes were grouped based on fluorescence intensity and methylene blue staining. A binary logistic regression model was established using 285 lymph node groups to predict metastatic risk. A total of 475 lymph nodes were identified, with 33 being metastatic. The metastatic risk reached 70% for partially blue-stained and weakly fluorescent lymph nodes between 1-2 cm. The model revealed associations between lymph node size, dye staining patterns, and metastatic risks (P<0.05). The AUC of the ROC curve was 0.855. The staining pattern of combined methylene blue and indocyanine green could predict risks of sentinel lymph node metastasis and facilitate rapid intraoperative identification of high-risk lymph nodes.

Deprivation of loading during rat Achilles tendon healing affects extracellular matrix composition and structure, and reduces cell density and alignment.

Tendon healing involves mechanosensitive cells that adapt to mechanical stimuli through mechanotransduction, resulting in increased tissue strength. However, detailed insights into this process in response to different loads remain limited. We aimed to investigate how different loading regimes impact the spatial composition of elastin and collagens during Achilles tendon healing. Histological analysis was conducted on healing rat Achilles tendons exposed to (1) full loading, (2) reduced loading, or (3) minimal loading. Histological analysis included Hematoxylin & Eosin and immunohistochemical staining targeting elastin, Collagen1, Collagen3, and CD31. Our results showed that the impact of mechanical stimuli on healing tendons varied with the degree of loading. Unexpectedly, minimal loading led to higher staining intensity for collagens and elastin. However, tendons exposed to minimal loading appeared thinner and exhibited a less organized matrix structure, with fewer, less aligned, and more rounded cells. Additionally, our findings indicated an inverse correlation between angiogenesis and load level, with more blood vessels in tendons subjected to less loading. Tissue integrity improved by 12 weeks post-injury, but the healing process continued and did not regain the structure seen in intact tendons even after 20 weeks. This study reveals a load-dependent effect on matrix alignment, cell density, and cell alignment.

The Association of Interleukin-36 Staining Intensity and Response to Biologic Therapy in Patients With Psoriasis: A Retrospective Immunohistochemical and Chart Review Pilot Study.

There are limited surrogate biomarkers to identify the active inflammatory pathway in psoriasis to direct treatment with targeted biologic therapies. We investigated the association of interleukin (IL)-36 epidermal expression, a diagnostic marker of psoriasis, with response to biologic therapy in patients with psoriasis. Retrospective immunohistochemical and chart review pilot study. Patients with psoriasis with low (scores 0-2) vs. high (scores 3-4) IL-36 expression did not have significantly different response rates to tumor necrosis factor α (TNFα), IL-17, and IL-12/23 or IL-23 inhibitors; and similarly, mean IL-36 expression scores did not significantly differ among responders vs. non-responders to each treatment mechanism. However, in patients with psoriasis treated with IL-12/23 or IL-23 inhibitors, there was a marked absolute difference in response rates in those with high vs. low IL-36 (84% vs. 50%, p = 0.12) and in mean IL-36 scores in responders vs. non-responders (3.35 vs. 2.57, p = 0.19). Patients with psoriasis with high IL-36 expression were more likely to respond to IL-12/23 and IL-23 inhibition than those with low IL-36, though these findings were not statistically significant. Additional studies with larger sample sizes are needed to validate and expand upon these findings.

TGF-β effects on adipogenesis of 3T3-L1 cells differ in 2D and 3D cell culture conditions.

The TGF-β superfamily plays a pivotal role in the regulation of adipogenesis, but little is known about the potential differential role of the three isoforms of TGF-β, TGF-β-1~3. To further elucidate their role, two-dimensionally (2D) and three-dimensionally (3D) cultured 3T3-L1 mouse preadipocytes were subjected to the following analyses: (a) qPCR analysis of adipogenesis-related factors and major extracellular matrix protein (2D and /or 3D), (b) lipid staining by Oil Red O (2D) or BODIPY (3D), (c) Seahorse cellular metabolic measurement (2D), and (d) size and stiffness measurements of 3D 3T3-L1 spheroids. In the 2D cultured 3T3-L1 cells, mRNA expression levels of adipogenesis-related genes and Oil Red O lipid staining intensity were significantly increased by adipogenesis and they were substantially decreased following treatment with 0.1 nm TGF-β isoforms, with TGF-β2 having the greater effects. Consistent with these results, treatment with TGF-β2 resulted in suppression of mitochondrial and glycolytic functions in 2D cultured 3T3-L1 cells. However, the inhibitory effect of TGF-β on adipogenesis decreased under 3D spheroid culture conditions and TGF-β isoforms did not affect adipogenesis-induced (a) enlargement and downsizing of 3T3-L1 spheroids, (b) increase in BODIPY lipid staining intensity, and (c) up-regulation of the mRNA expression of adipogenesis-related genes. The findings presented herein suggest that the three TGF-β isoforms have different suppressive effects on adipogenesis-related cellular properties of 2D cultured 3T3-L1 cells and that their effects decrease under 3D spheroid culture conditions.

A COMPARATIVE STUDY OF CLINICOHISTOPATHOLOGICAL CORRELATION IN PATIENTS OF PSORIASIS AND PSORIASIFORM LESIONS WITH KI67 AND CD34 IMMUNOHISTOCHEMICAL EXPRESSION

Objectives: Psoriasis, characterized by chronic inflammation of the skin, stands as a notable example of a psoriasiform tissue pattern. In dermatology, the term “psoriasiform dermatitis” frequently appears across various inflammatory skin conditions, presenting challenges for dermatologists and pathologists alike in accurate differentiation. Relying solely on clinical features may prove insufficient in distinguishing between psoriasis and other psoriasiform dermatoses. Therefore, comprehensive differentiation requires consideration of clinical, histopathological, and immunohistochemical factors. Our study is to compare the clinicohistopathological features of psoriasis and psoriasiform dermatitis and to evaluate the immunohistochemical expression of CD34 and Ki67 in psoriasis and psoriasiform dermatitis. Methods: This retrospective study was conducted at the Department of Pathology, Mediciti Institute of Medical Sciences, Ghanpur, focusing on cases between January 1, 2020, and December 31, 2022. Clinical histories were extracted from requisition forms submitted by the Department of Dermatology. The study analyzed paraffin blocks from 50 patients each diagnosed with psoriasis and psoriasiform dermatitis. Formalin-fixed paraffin-embedded tissue samples were processed into 4-micron sections and initially stained with hematoxylin and eosin. The immunohistochemical analysis included primary antibodies against Ki-67 and CD34, with positive controls sourced from strongly positive samples of squamous cell carcinoma and capillary hemangioma, respectively. Appropriate negative controls were employed to ensure the accuracy of our findings. Results: In our study, we analyzed a total of 100 cases, comprising 50 cases each of psoriasis and psoriasiform dermatitis. Among the psoriasis cases, there was a male predominance with 29 males and 21 females, while psoriasiform dermatitis also showed male predominance with 32 males. The mean age was 33.8 years for psoriasis and 34.8 years for psoriasiform dermatitis. The majority of psoriasis cases fell in the 21–30 years age range, whereas for psoriasiform dermatitis, it was between 31 and 40 years. Significantly, our immunohistochemical analysis revealed that the mean staining intensity of the two markers was notably higher in psoriasis compared to psoriasiform dermatitis. Conclusions: In our study, we observed that the expression levels of Ki-67 and CD34 biomarkers were significantly elevated in psoriasis compared to psoriasiform dermatitis. Notably, four cases of psoriasiform dermatitis demonstrated positivity for CD34 in our analysis. This finding suggests a potential avenue for further research to explore the implications of CD34 positivity in psoriasis.

Application of mitoxantrone hydrochloride in lymph node imaging during radical thyroidectomy

Objective: To investigate the clinical application value of the new tracer mitoxantrone hydrochloride in imaging of cervical lymph nodes and identifying of parathyroid, recurrent laryngeal nerve during thyroid cancer radical surgery. Methods: A prospective controlled study was conducted between January 2022 and April 2024 at Tianjin First Central Hospital, recruiting 180 patients with thyroid cancer, including 54 males and 126 females, aged from 26 to 69 years. The patients were randomly divided into three groups: the mitoxantrone hydrochloride lymph node tracing group (MHI group), the nanocarbon lymph node tracing group (nanocarbon group), and the control group without lymph node tracers, with 60 cases in each group. All patients underwent total thyroid resection and regional lymph node dissection. The number of detected lymph nodes and positive metastatic lymph nodes and surgical field clarity scores and the identification rate of parathyroid glands and the instances of inadvertent excision were compared between the groups. Serum calcium and parathyroid hormone (PTH) levels at different perioperative time points and treatment-related complications were analyzed. One-way ANOVA, chi-square test and two independent sample non-parametric tests, were employed for statistical analyses. Results: The mean operation time, neck drainage volume and hospital stay in the MHI group and the nanocarbon group were respectively significantly lower than those in the control group (P<0.05). The mean staining intensity score, tracer success rate and surgical field clarity score in MHI group were respectively significantly higher than those in nanocarbon group (P<0.05). The mean numbers of dissected lymph nodes positive metastatic nodes and identification rates of parathyroid glands in MHI group were respectively significantly higher than those in both the nanocarbon and control groups (P<0.05). The rates of inadvertent parathyroid excision in both MHI group and nanocarbon group were significantly lower than those in control group (P<0.05). On postoperative day 1 and day 5, mean serum calcium and PTH levels in MHI group were respectively significantly higher than those in nanocarbon group and control groups. However, on postoperative day 1, there was no difference in mean serum calcium levels between the nanocarbon group and the control group (P>0.05), though PTH level in the nanocarbon group was higher than that in the control group. By postoperative day 5, both serum calcium and PTH levels were higher in the nanocarbon group compared to the control group (P<0.05). On postoperative days 30, there were no differences in serum calcium and PTH levels between the MHI group and the nanocarbon group (P>0.05). The risks of facial numbness, hand and foot tetany and choking during drinking were lower in both the MHI and nanocarbon groups compared to the control group (P<0.05). Conclusion: MHI demonstrates advantages in rapid targeted delivery and clear staining of the surgical field during regional lymph node dissection in radical thyroidectomy for thyroid cancer, effectively reducing the risk of collateral damage to the recurrent laryngeal nerve and parathyroid glands.

Chitosan-glucose derivative membrane obtained by Maillard reaction improves cartilage repair in a rabbit model

BackgroundTreatment of articular cartilage injury remains a challenging clinical problem in orthopedics. Chitosan-derived biomaterial could be a potential adjuvant treatment to improve cartilage repair. In the current study, we examined the effects of two potential chitosan-derived materials on cartilage regeneration of osteochondral defects in rabbits.MethodsAn osteochondral defect was created over the rabbit knee and treated using three approaches: group A received no material (n = 24), group B received chitosan membranes with glucose absorption (CGA; n = 25), and group C received chitosan-glucose derivative membranes obtained via the Maillard reaction (CGMR; n = 25). Cartilage repair over the osteochondral defect was analyzed 12 weeks post-surgery via histological analysis, immunostaining, and reverse transcription-qualitative polymerase chain reaction (RT-qPCR) for type-I and type-II collagen mRNA.ResultsAccording to histological analysis, CGMR-treated defects showed significantly improved modified O’Driscoll scoring when compared with no material- and CGA-treated defects (20.9 ± 4.3 vs. 13.00 ± 2.5 and 17.7 ± 4.6, p < 0.001). Moreover, group C exhibited higher intensity of type-II collagen immunohistochemical staining over the regenerated cartilage than groups A and B, along with increased expression of type-II collagen mRNA by RT-qPCR.ConclusionsCGMR might improve cartilage regeneration in osteochondral defects.

β‑catenin expression in endometrioid type endometrial cancer: Expression patterns and impact on disease outcomes.

Determination of nuclear and/or cytoplasmic expression of β-catenin by immunohistochemistry in patients with endometrial cancer (EC) may constitute a potential diagnostic method for identifying patients with a catenin β1 (CTNNB1) gene mutation and those at risk of disease recurrence. The present study aimed to investigate β-catenin expression patterns in hysterectomy specimens of patients with endometrioid type EC using immunohistochemistry, and to examine the prognostic impact of β-catenin. The study was a single-institutional, retrospective cohort trial enrolling consecutive patients with a postoperative histopathological diagnosis of endometrioid EC who underwent hysterectomy between January 2015 and December 2018. Histopathology slides from 75 patients were stained with a monoclonal antibody targeting the β-catenin protein. Any percentage of nuclear staining, whether focal or diffuse, was considered 'β-catenin nuclear-positive'. The cytoplasmic staining reaction of β-catenin was assessed based on the percentage of stained cells and staining intensity. Immune-reactivity score (IRS) values were determined by multiplying the scores for the percentage of staining and staining intensity. IRS values 0 to 2 were regarded as negative expression, 3 to 4 as low expression, 6 to 8 as moderate expression, and 9 to 12 as high expression. Recurrence-free survival (RFS) was used as the prognostic endpoint. Only 2 out of 75 tissue samples (2.7%) exhibited nuclear β-catenin expression, with a low staining percentage of 5%. By contrast, cytoplasmic staining was observed in all samples (100%). According to the IRS findings, 1.3% of the samples exhibited negative cytoplasmic expression, 42.7% low expression, 38.7% moderate expression and 17.3% high expression. Cox regression analysis revealed that staining with β-catenin, either nuclear or cytoplasmic, had no impact on RFS, and stage was the sole independent prognostic factor. In conclusion, based on these results, β-catenin expression in endometrioid EC was revealed to be mostly cytoplasmic, with only 2.7% of tissue samples exhibiting nuclear expression. Overall, β-catenin expression has no impact on RFS.

Application of carbonic anhydrase IX in sporadic hemangioblastoma of the central nervous system and hemangioblastoma associated with von Hippel-Lindau disease.

This research aims to examine the expression of carbonic anhydrase IX (CAIX) protein in hemangioblastoma of the central nervous system and its potential application in pathological diagnosis and differential diagnosis. Immunohistochemistry was used to identify the expression of CAIX and the α-inhibin protein. The sensitivity and specificity of CAIX and α-inhibin for identifying hemangioblastoma of the central nervous system were compared. In addition, 86 patients with meningiomas were gathered to detect CAIX protein expression. Hemangioblastoma and angiomatous, microcystic the two subtypes of meningiomas, were compared for CAIX and EMA protein expression. In hemangioblastoma, there were significant differences in the median positive percentage and staining intensity of CAIX and α-inhibin (p < 0.05). There was no discernible difference in the expression of the CAIX protein between sporadic hemangioblastoma of the central nervous system and those linked to von Hippel‒Lindau disease. In comparison to angiomatous and microcystic meningiomas, the positive rate of CAIX in hemangioblastomas was substantially greater (p < 0.001). The expression of EMA in microcystic meningioma (6/6) and angiomatous meningioma (17/17) was significantly different from hemangioblastoma (0/30) (p < 0.0001). Hemangioblastoma might be diagnosed with high specificity and sensitivity through CAIX immunohistochemistry. The combination of CAIX with EMA is useful for the diagnosis and differential diagnosis of hemangioblastoma.

Predictive Value of E-Cadherin, β-Catenin and SOX4 in Recurrence and/or Progression of Non-Muscle Invasive Urinary Bladder Transitional Cell Carcinoma. A Retrospective Study

Abstract Background Bladder cancer (BC) is one of the most common genitourinary cancers worldwide, the pathophysiology and etiology of BC are complicated, including both internal genetic and environmental factors. Currently, some studies suggest that the abnormal function of intercellular adhesion molecules is highly correlated with the invasion of surrounding tissues and distant organ metastasis of BC. Objective To develop a correlation between these markers and NMIBC and to help predict the recurrence and/or progression. Patients and Methods It is a retrospective cohort (observational) study assessing a correlation regarding the alternation in the expression of E-cadherin, β-catenin and SOX4 in determining the recurrence and/or progression and their prognostic value in NMIBC TCC. Patients Diagnosed with NMIBC TCC and their follow ups data were evaluated at the urology department in Theodor Bilharz Research Institute (TBRI) and Ain Shams University from January 2019 – till July 2022. Results Loss of normal expression of E-cadherin and β-catenin, upregulation of SOX4 is significantly correlated with BC and tumorigenesis, yet no clear relation between E-cadherin, β- catenin and SOX4 in progression or recurrence of NMIBC, regarding It’s staining intensity, percentage of positive cells and immunoreactive score (IRS). Conclusion bases on our current study, E-cadherin, β-catenin and SOX4 showed highly significance difference between normal urothelium and TCC NMIBC indicating it may have a role in diagnosis, although its value changes with follow up of the patients but of non-significant difference to predict prognosis, recurrence and/or progression among TCC NMIBC.

Loss of phosphatase and tensin homolog expression castration-sensitive prostate cancer predicts outcomes in men after prostatectomy.

This study aimed to investigate the potential for using the phosphatase and tensin homolog (PTEN) gene as a prognostic marker in post-prostatectomy patients with castration-sensitive prostate cancer (PCa). A total of 180 patients with castration-sensitive PCa who underwent radical prostatectomy at our institution were included in this study. PTEN expression was evaluated using immunohistochemistry, and patients were classified into two groups based on the staining intensity: PTEN-Normal and PTEN-Loss. The association between PTEN expression and biochemical recurrence was analyzed using the Cox proportional hazards model. Patients in the PTEN-Loss group had a higher risk of biochemical recurrence (hazard ratio, 4.642; 95% confidence interval, 2.137-10.083; p < 0.001) and a lower recurrence-free rate compared to the PTEN-Normal group (35% vs. 75%). In addition to clinicopathological factors, such as the serum prostate-specific antigen level, Gleason score, and T stage, evaluation of PTEN expression improved the prediction of biochemical recurrence after prostatectomy (area under the curve, 0.577 vs. 0.688). Low PTEN expression is a significant predictor of biochemical recurrence in patients with castration-sensitive PCa who have already undergone prostatectomy.

Evaluation of Tumour-Associated Macrophages and Colony-Stimulating Factor-1 Expression in Invasive Breast Carcinoma and Their Association with Prognostic Parameters

INTRODUCTION: Recent breast cancer research has focused on tumour microenvironment (TME). Tumour-associated macrophages (TAMs) are the key players in TME as they provide pro-tumorigenic milieu for tumour progression and metastasis. These macrophages are primarily regulated by colony-stimulating factor-1 (CSF-1) secreted by breast cancer cells. This study investigated the association of localization of TAMs infiltration within breast carcinoma and CSF-1 expression by cancer cells with the pathological prognostic parameters. MATERIALS AND METHODS: TAMs were assessed in 128 cases of invasive breast carcinoma by CD163 immunohistochemical expression. The median TAM density in both the tumour nest and tumour stroma was utilized to classify TAMs into categories of low and high infiltration. The cancer cells were immunostained with anti-CSF-1 antibody and the staining intensity was evaluated as low or high expression. RESULTS: High nest and stromal TAMs were associated with higher tumour grades (p=0.005 and p=0.0001, respectively) whereas only high stromal TAMs showed significant association with negative oestrogen and progesterone receptors status (p=0.001 and 0.001, respectively); and triple-negative subtype (p=0.002). High CSF-1 expression was significantly associated with high stromal TAMs (p=0.031). High CSF-1 expression was associated with tumour grade and positive HER2 status (p=0.008 and 0.007, respectively). CONCLUSION: TAMs in tumour nest and stroma showed varying degrees of association with the clinicopathological parameters. High CSF-1 expression was associated with unfavourable prognostic parameters. Therefore, the evaluation of TAMs and CSF-1 expressions could potentially serve as prognostic markers and cellular targets for novel treatment modality in invasive breast cancers.

Characterisation of the tumour microenvironment in primary and recurrent glioblastomas.

Glioblastoma patients have a dismal prognosis, due to inevitable tumour recurrence and respond poorly to immunotherapy. Tumour-associated microglia/macrophages (TAMs) dominate the glioblastoma tumour microenvironment and have been implicated in tumour progression and immune evasion. Early recurrent glioblastomas contain focal reactive regions with occasional fibrosis, chronic inflammation, TAMs and tumour cells. Surgical specimens from these tumours are rare and provide crucial insights into glioblastoma recurrence biology. This study aimed to characterise TAM- and lymphocyte phenotypes in primary vs early- and late-recurrent glioblastomas. Patient-matched primary and recurrent glioblastomas were compared between patients with early recurrences (n = 11, recurrence ≤6months) and late recurrences (n = 12, recurrence after 12-19 months). Double-immunofluorescence stains combining Iba1 with HLA-DR, CD14, CD68, CD74, CD86, CD163, CD204 and CD206 along with stains for CD20, CD3, CD8 and FOXP3 were quantified with software-based classifiers. Reactive regions in early recurrent tumours contained more TAMs (31.4% vs 21.7%, P = 0.01), which showed increased expression of CD86 (59.4% vs 38.4%, P = 0.04), CD204 (48.5% vs 28.4%, P = 0.03), CD206 (25.5% vs 14.4%, P = 0.04) and increased staining intensity for CD163 (86.4 vs 57.7 arbitrary units, P = 0.02), compared to late recurring tumours. Reactive regions contained more B-lymphocytes compared to patient-matched primary tumours (0.71% vs 0.40%, P = 0.04). Fractions of total, cytotoxic and regulatory T-lymphocytes did not differ. Early recurrent glioblastomas showed enrichment for TAMs, expressing both pro- and anti-inflammatory markers and B-lymphocytes. This may indicate a time-dependent response to immunotherapy explained by time-dependent alterations in the immune-microenvironment in recurrent glioblastomas.

Genetic Signatures: CD44 Single-Nucleotide Polymorphisms Affect Cell Surface Expression and Elevate Risk in Head and Neck Squamous Cell Carcinoma.

This study aimed to investigate the impact of single-nucleotide polymorphisms (SNPs) in the CD44 gene, specifically in the 3'UTR region (rs13347) and intronic region (rs187115), on the cell surface expression of CD44 protein and the risk of development of head and neck squamous cell carcinoma (HNSCC). The study involved analysis of 85 samples and 85 healthy controls. Immunohistochemistry (IHC) and flow cytometry were used to assess cell surface protein expression using CD44 antibody. DNA from formalin-fixed paraffin-embedded tissue sections was isolated and amplified using targeted primers. Sanger sequencing of the resultant amplified products was performed to determine the genotypes of the CD44 rs13347 and rs187115 SNPs. GTEx and RegulomeDB were queried to evaluate the genotypic effects of these variants on target gene expression and regulation. A comparison between patients with HNSCC and healthy controls revealed a significant association between CD44 rs13347 and an increased risk of HNSCC in all the analyzed models, especially the TT genotype showed a significantly higher risk with an odds ratio of 8.69 (95% CI, 2.35 to 32.09; P = .0003). However, no significant association was found between CD44 rs187115 and HNSCC in any of the models analyzed (all P > .05). Other notable findings included significant associations between CD44 rs13347 genotype and age (P = .031), number of CD44-positive tumor cells (P = .049), CD44 staining intensity (SI; P = .039), and CD44 immunoreactivity score (IRS) status (P = .019). The T allele and homozygous TT genotype of CD44 rs13347 SNP were associated with increased susceptibility to HNSCC and decreased proportion of CD44-positive tumor cells, low SI, and reduced IRS.

Immunohistochemical Expression of P63 and P40 in Squamous Cell Carcinoma of Head and Neck

Background: Squamous cell carcinoma (SCC) in head and neck region has a high worldwide prevalence. Increased accuracy in diagnosis is achieved by immunohistochemical markers. Objectives: To evaluate the immunohistochemical expression of p63 and p40 in SCC and correlate with histological grades to determine p40 as a better potential diagnostic immunohistochemical marker for SCC. Methods: Descriptive cross-sectional study was conducted on eighty-three cases of SCC of head and neck from March 2020 to February,2022 at Pathology Department, KEMU, Lahore. Data including age, sex, risk factors and tumor site was collected. SCC grading was done on H&amp;E stained tissue sections. Staining with p63 and p40 was performed and independently scored for percentage of tumor cells stained and intensity of nuclear staining. The results were compared using H-scores in all grades of SCC. Data was entered in Statistical Package for Social Sciences(SPSS-26). Quantitative variables like age were presented as mean ±SD. Qualitative variables as frequency and percentages. Correlation of two groups was done by applying Chi-square test. P value ≤0.05 was taken as significant. Results: SCC was graded according to the W.H.O. grading system. All cases stained positive for p63 and p40. Strong nuclear staining by p40 was observed in 65.02% cases versus 42.17% by p63. The mean tumor area stained by p40 was greater than p63 (90.35% vs 75.5%) in all cases. A significant higher mean H-score for p40 (9.69±2.86) versus p63 (7.05±3.12) was observed as indicated by significant P-value. The mean H-score of p40 immunoreactivity was 10.11 and p63 was 6.00 in poorly differentiated SCC. Males comprised 74.70% cases whereas females 25.30% only. SCC in oral cavity was (96.39%) followed by oropharynx and larynx. Conclusion: P40 immunohistochemical marker was better compared to P63 for the diagnosis of SCC particularly in poorly differentiated SCC.

Aberrant positivity for BCOR immunohistochemistry in merkel cell carcinoma - a potential diagnostic pitfall

BackrgoundMerkel cell carcinoma (MCC) is a rare, aggressive primary cutaneous neuroendocrine carcinoma, frequently associated with clonal Merkel cell polyomavirus integration. MCC can pose significant diagnostic challenges due to its diverse clinical presentation and its broad histological differential diagnosis. Histologically, MCC presents as a small-round-blue cell neoplasm, where the differential diagnosis includes basal cell carcinoma, melanoma, hematologic malignancies, round cell sarcoma and metastatic small cell carcinoma of any site. We here report strong aberrant immunoreactivity for BCOR in MCC, a marker commonly used to identify round cell sarcomas and other neoplasms with BCOR alterations.MethodsBased on strong BCOR expression in three index cases of MCC, clinically mistaken as sarcoma, a retrospective analysis of three patient cohorts, comprising 31 MCC, 19 small cell lung carcinoma (SCLC) and 5 cases of neoplasms with molecularly confirmed BCOR alteration was conducted. Immunohistochemical staining intensity and localization for BCOR was semi-quantitatively analyzed.ResultsThree cases, clinically and radiologically mimicking a sarcoma were analyzed in our soft tissue and bone pathology service. Histologically, the cases showed sheets of a small round blue cell neoplasm. A broad panel of immunohistochemistry was used for lineage classification. Positivity for synaptophysin, CK20 and Merkel cell polyoma virus large T-antigen lead to the diagnosis of a MCC. Interestingly, all cases showed strong positive nuclear staining for BCOR, which was included for the initial work-up with the clinical differential of a round cell sarcoma. We analyzed a larger retrospective MCC cohort and found aberrant weak to strong BCOR positivity (nuclear and/or cytoplasmic) in up to 90% of the cases. As a positive control, we compared the expression to a small group of BCOR-altered neoplasms. Furthermore, we investigated a cohort of SCLC as another neuroendocrine neoplasm and found in all cases a diffuse moderate to strong BCOR positivity.ConclusionsThis study demonstrates that neuroendocrine neoplasms, such as MCC and SCLC can express strong aberrant BCOR. This might represent a diagnostic challenge or pitfall, in particular when MCC is clinically mistaken as a soft tissue or a bone sarcoma.

Expression of Programmed Death Ligand 1 and Indoleamine 2,3-Dioxygenase in Oral Lichen Planus and Oral Lichenoid Lesions.

Oral lichen planus (OLP) and oral lichenoid lesions (OLL) are inflammatory T-cell mediated disorders of the oral mucosa (OM). Both are associated with an increased risk of oral squamous cell carcinoma, with OLL possibly having a higher rate of malignant transformation than OLP. Programmed death ligand 1 (PD-L1) and indoleamine 2,3-dioxygenase (IDO) are immunosuppressive molecules possessing inhibitory effect on T-cells and have been implicated in carcinogenesis. The aim of this study was to examine the expression of PD-L1 and IDO in OLP and OLL. Sixty-eight formalin-fixed, paraffin-embedded tissue samples diagnosed as OLP, compatible with OLP, or OLL were divided into OLP (n = 39) or OLL (n = 29) groups based on both clinical and histopathological diagnostic criteria. Samples of healthy OM (n = 9) served as controls. Samples were immunohistochemically stained for PD-L1 and IDO, and staining distribution and intensity were evaluated. Immunohistochemical expression of PD-L1 was increased in the basal and intermediate layers of epithelium in OLP and in lamina propria in both OLP and OLL compared to controls. OLP and OLL showed increased expression of IDO in epithelium and lamina propria compared to controls. PD-L1 staining intensity in the basal epithelial layer, and IDO staining intensity in lamina propria were increased in OLP compared to OLL. The results indicate that the expression of PD-L1 and IDO increases in OLP and OLL, suggesting that these molecules may play a role in the pathogenesis of both disorders.

EXPRESSION OF VITAMIN D3 RECEPTOR IN TUMOR TISSUE AND RELATIONSHIP BETWEEN CLINICAL FEATURES, PROGNOSTIC INDICATORS AND EXPRESSION OF VITAMIN D3 RECEPTOR IN HODGKIN LYMPHOMA

&lt;p&gt;&lt;strong&gt;Background and objectives:&lt;/strong&gt; In our study, we aimed to investigate the expression of vitamin D receptor and the relationship between clinical features, prognostic indicators, and expression of vitamin D receptor in Hodgkin lymphoma. &lt;strong&gt;Materials and Methods:&lt;/strong&gt; Pathology records and medical files of patients with Hodgkin lymphoma were evaluated. Forty-two tissue samples were analyzed retrospectively. Vitamin D receptor staining was performed with immunohistochemistry. The patients were divided into low, medium, and intense staining subgroups. The demographic data, clinical and biochemical values of patients were reviewed retrospectively, and the relationship between clinical features, prognostic indicators, and vitamin D receptor expressions were examined. &lt;strong&gt;Results:&lt;/strong&gt; VDR staining was positive in Reed-Sternberg cells of all Hodgkin lymphoma patients. The cases were divided into three groups according to VDR staining intensity as low, medium and intense staining and compared with known clinical prognostic parameters. Only the presence of symptom B was statistically significantly different. Gender, bone marrow involvement, extralymphatic involvement, mediastinal mass, bulky mass, mean leukocyte count, and treatment response were associated with vitamin D receptor staining intensity, but they were statistically insignificant. &lt;strong&gt;Conclusion: &lt;/strong&gt;Our study is the second study showing VDR in tumor Hodgkin lymphoma tumor tissues and is meant to be the first to evaluate its association with clinical and biochemical prognostic laboratory tests. Vitamin D receptor staining in all Hodgkin lymphoma patients led to the conclusion that VDR can be used in the histopathological differential diagnosis of HL. More studies are needed to assess and demonstrate the relationship between prognostic and diagnostic indicators and vitamin D receptor intensity of immunohistochemically detected VDR in tumor tissues of Hodgkin lymphoma. Also, further studies with serum vitamin D levels and other histologic subtypes can provide more reliable results.&lt;/p&gt;&lt;p&gt; &lt;/p&gt;&lt;p&gt;&lt;strong&gt; Article visualizations:&lt;/strong&gt;&lt;/p&gt;&lt;p&gt;&lt;img src="/-counters-/soc/0715/a.php" alt="Hit counter" /&gt;&lt;/p&gt;

Fabrication of 3D microfibrous composite polycaprolactone/hydroxyapatite scaffolds loaded with piezoelectric poly (lactic acid) nanofibers by sequential near-field and conventional electrospinning for bone tissue engineering

Near-field electrospinning (NFES) has recently gained considerable interest in fabricating tissue engineering scaffolds. This technique combines the advantages of both 3D printing and electrospinning. It allows for the production of fibers with smaller resolution and the ability to make regular structures with suitable pores. In this study, a microfibrous composite scaffold of polycaprolactone (PCL)/hydroxyapatite (HA) was prepared by NFES in the first step. The microfibrous scaffold had a fiber spacing of 414.674 ± 24.9 μm with an average fiber diameter of 94.695 ± 16.149 μm. However, due to the large fiber spacing, the surface area was insufficient for cell adhesion. Therefore, the hybrid scaffold was prepared by adding aligned and random electrospun poly (L-lactic acid) (PLLA) nanofibers to the microfibrous scaffold. Cellular studies showed that cell adhesion to the hybrid scaffold increased by 334 % compared to the microfibrous scaffold. These nanofibers also exhibited piezoelectric properties, which helped stimulate bone regeneration. Aligned nanofibers in the hybrid scaffold enhanced alkaline phosphatase activity and the intensity of alizarin red staining 1.5 and 1.6 times, respectively, compared to the microfibrous scaffold. Furthermore, the elastic modulus and ultimate tensile strength increased by 268 % and 130 %, respectively, by adding aligned nanofibers to the microfibrous scaffold. Therefore, the hybrid microfibrous composite scaffold of PCL/HA containing aligned electrospun PLLA nanofibers with improved properties showed the potential for bone regeneration.