Bcl-2 Staining Research Articles

Deciphering the spectrum of cutaneous lymphomas expressing TFH markers

T-follicular helper (TFH) markers are expressed in the microenvironnement of marginal zone B-cell lymphoma (MZL), and in lymphomas arising from TFH-cells, sometimes making the differential diagnosis difficult. In the skin, the “TFH-spectrum” is poorly defined, going from primary cutaneous lymphoproliferative disorder with small/medium CD4+ T-cells (SMLPD) to cutaneous localizations of systemic angioimmunoblastic T-cell lymphoma (cAITL), and may pass through intermediate forms (primary cutaneous T-follicular helper derived lymphoma, not otherwise specified (PCTFHL,NOS)). We retrospectively analyzed 20 MZL, 13 SMLPD, 5 PCTFHL, and 11 cAITL clinically, histologically, and molecularly, to define tools to differentiate them. Characteristics that might favor the diagnosis of MZL over SMLPD are: multiple skin nodules (p < 0.001), nodular architecture (p < 0.01), residual germinal centers with follicular dendritic cell network (p < 0.001), monotypic plasma cells (p < 0.001), and few staining with PD1 (p = 0.016) or CXCL13 (p = 0.03). PCTFHL and cAITL presented as multiple (p < 0.01) lesions, in older patients (p < 0.01), with systemic symptoms and/or biological alterations (p < 0.01). Immunophenotypic loss of T-cell markers (p < 0.001), BCL6 (p = 0.023) and/or CD10 staining (p = 0.08), and a higher proliferative index (≥ 30%, p = 0.039) favoured these diagnoses over SMLPD. Pathogenic variants were observed by genomic sequencing in 47% of MZL (TNFAIP3 (32%), EP300 (21%), NOTCH2 (16%), KMT2D (16%), CARD11 (10.5%)), 8% of SMLPD (TET2), 40% of PCTFHL (SOCS1 (20%), ARID1A (20%)) and 64% of cAITL (TET2 (63.6%), RHOA (36.4%), NOTCH1 (9%)). This study characterizes the various clinical and histological features between cutaneous lymphomas expressing TFH markers and highlights the value of the interest of screening for genomic mutations in difficult cases.

Construction of Fe-doped ZIF-8/DOX nanocomposites for ferroptosis strategy in the treatment of breast cancer.

Breast cancer has become one of the top five commonest causes of cancer death. The use of ferroptosis to induce the generation of reactive oxygen species (ROS) in cancer cells presents a promising and potential strategy for cancer treatment. Herein, a series of facile bimetallic nanoparticles (x% Fe-doped ZIF-8) were synthesized and tested, and doxorubicin (DOX), a classic drug for breast cancer therapy, was encapsulated. After comparing the ratios of Fe2+/(Fe2+ + Zn2+), 7% Fe-doped ZIF-8 (7FZ) was found to be the most suitable particle for medical application. The drug loading efficiency of DOX@7FZ was 58.01 ± 0.02%. The pH-sensitive DOX@7FZ was degraded and DOX was released in lysosomes once internalized. Both the intracellular content of iron and ROS increased significantly. Meanwhile, the cell viability declined to 13.98% in 24 h at a concentration of 60 μg mL-1 and the IC50 was 42.68 μg mL-1. Moreover, the expression of Bcl-2 and GPX-4 proteins decreased in a time-dependent manner, indicating that DOX@7FZ was able to enhance the ROS level in cancer cells via a synergistic effect between apoptosis and ferroptosis. The mechanism of action of DOX@7FZ was further verified using hematoxylin and eosin staining and immunohistochemical staining of Bcl-2 and GPX-4. These remarkable characteristics of DOX@7FZ may inspire further advancements in the treatment of breast cancer.

Primary cutaneous blastic marginal zone lymphoma: A comprehensive clinical, light microscopic, phenotypic and cytogenetic appraisal

BackgroundPrimary cutaneous marginal zone lymphoma (PCMZL) is a form of indolent lymphoproliferative disease where the disease is largely a cutaneous confined process. It is typically a neoplasm composed of post germinal small B-cells and light chain restricted plasma cells in a background of reactive T-cell hyperplasia and benign germinal centers. Rarely a significant degree of large cell infiltration occurs warranting the categorization as blastic marginal zone lymphoma. Materials and methodsWe reviewed our data base over a time period of 2016 to 2022 for cases diagnosed as blastic MZL. Twelve cases were identified. The clinical records and pathological data were reviewed. ResultsNine of the cases represented de novo forms of blastic MZL while in three cases there was a prior history of MZL. Multifocal cutaneous disease was not uncommon and one quarter of the cases had evidence of extracutaneous dissemination. All patients except three achieved remission with varied therapeutic interventions depending on the extent of the disease ranging from conservative re-excision to chemotherapy. No patient died from lymphoma. Light microscopically, there was evidence of a background of conventional MZL in the majority of cases. The large cell component was typically characterized by multiple micronodular aggregates throughout the dermis although in three cases there was a striking diffuse large cell component as the dominant infiltrate. Phenotypically, a third of the cases showed either CD5 or CD23 positivity amidst neoplastic B cells. Significant staining for BCL-2 was noted in the majority of cases tested while extensive MUM-1 positivity was observed in half of the cases tested. Kappa or lambda light chain restriction was seen in most. The Ki67 proliferation index exceeded 30 % in all cases. There was C-MYC positivity in two cases. While most cases did not detect cytogenetic abnormalities, one case had multiple cytogenetic hits that are associated with diffuse large B cell lymphoma. Next generation sequencing showed a Ten-eleven translocation 2 mutation in the earlier biopsy prior to transformation and in the later biopsy after transformation along with an additional B2M mutation in the transformed biopsy. Both types of mutations are very uncommon but held to contribute to tumor progression in the setting of diffuse large B cell lymphoma. ConclusionBlastic MZL is associated with a more aggressive clinical course. Even when there is disseminated disease patients while not always cured did not have a fatal course in this series. The light microscopic findings are reproducible. The background of MZL, identification of larger cells in significant numbers without a follicle center phenotype, at times expressing CD5 or CD23 with variable positivity for MUM1, BCL-2 and C-MYC and a high proliferation index define the pathology in most. Certain cytogenetic abnormalities and genetic mutations implicated in large cell transformation into a diffuse large B cell lymphoma are seen in blastic MZL with earlier biopsies prior to transformation potentially harboring at risk genetic mutations.

Clinical Laboratory Mutation Analysis Performed on Tumor Biopsies from Patients with Relapsed/Refractory Aggressive B Cell Lymphoma Treated with Palliative-Intent Therapy

Background While recently published analyses have characterized findings from comprehensive genomic analyses (PMID: 29713087, 29641966) or targeted sequencing (PMID: 32187361) performed on tumor biopsies (bx) from patients (pts) with newly-diagnosed diffuse large B cell/high grade B cell lymphoma (DLBCL/HGBL), analyses of mutations (mut) found in tumor bx from pts with relapsed/refractory (R/R) DLBCL/HGBL have been limited and largely descriptive in nature (PMID: 26647218, ICML 2017 #294). We aim to describe the clinicopathologic characteristics including tumor mutation profile, as well as therapy (tx) received and survival outcomes for pts with R/R DLBCL/HGBL who were treated with palliative-intent at our institution. Methods Pts diagnosed with DLBCL/HGBL who had at least one prior tx and experienced R/R disease between 2015-2021, who had tumor bx performed immediately prior to the receipt of palliative-intent tx which underwent clinical laboratory mutation analysis (CLMA) performed at the University of Pennsylvania with one of three lymphoma sequencing panels, were retrospectively analyzed. Tx was given at the discretion of treating physician. Result of fluorescence in situ hybridization for MYC, BCL2 and BCL6 as well as immunohistochemical staining for MYC and BCL2 were from current bx or prior bx if not performed on current bx. Overall survival (OS) was defined from time of bx of R/R disease and date of death or last follow-up while alive, with cut-off at 7/1/22. Univariate Cox regression analysis for OS from time of bx was performed. Results CLMA was performed on 35 pt tumor bx with 39 successful assays (86% success rate) with median turnaround time for successful result of 19.5 days. Baseline clinicopathologic and treatment characteristics can be seen in the Table. Sequencing panels identified CREBBP as the most frequent mutation present in the tissue samples, occurring in 12 (48%) of samples tested for CREBBP, followed by TP53 occurring in 9 (30%) and MYD88 occurring in 7 (23%). A histogram of all detected mutations with n≥1 is depicted in the Figure. Next-line of tx was classified as polatuzumab-containing (n=8), small molecule inhibitor-containing (7), tafasitamab-containing (4), bispecific antibody (n=3) and other (n=8). With a median length of follow-up of 19.1 months, the median OS for the entire population was 18.1 months. Characteristics listed in the Table which were predictive of OS with P<0.10 on univariate Cox regression analysis were International Prognostic Index Score at the time of relapse ≥3 vs <3 (hazard ratio [HR] 3.0, 95% confidence interval [CI] 0.85-10.3, P=0.09), HGBL vs DLBCL histology (HR 6.8, 95% CI 2.3-20.2, P=0.001), MYC rearrangement vs not (HR 2.4, 95% CI 0.9-6.6, P=0.08) and double hit lymphoma vs not (HR 4.8, 95% CI 1.6-14.5, P=0.005). None of the above-mentioned mut predicted for OS. Conclusions In this pt population with R/R DLBCL/HGBL receiving palliative-intent tx, mut identified by CLMA revealed CREBBP, TP53 and MYD88 occurred frequently these nor other mut predicted for OS. Although a relatively small sample population without uniform tx, this study demonstrates the presence of recurring mut which could be used to guide palliative tx decisions. A study with a larger cohort of pts whose tumor bx undergo CLMA with a single comprehensive mut panel could be valuable in further understanding the predictive value of tumor biopsy mut this pt population based on tx received. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Significance of Bcl-2 expression in breast cancer

Introduction and Aim: The most prevalent and lethal form of cancer in women, breast carcinoma is thought to account for 2,088, 849 (11.6%) of all new cases each year. Protooncogene Bcl-2 is primarily present in the perinuclear membrane. Examining the significance of Bcl-2 expression as a predictive factor in breast cancer is the goal of the current investigation. Materials and Methods: From December 2019 to January 2021, a tertiary care hospital in Chennai conducted this investigation on 42 cases of mastectomy specimens. Using tools from Path Insitu, Bcl-2 marker immunohistochemistry was carried out. Scores were taken after observing Bcl-2 expression in the cytoplasm of tumor cells. Results: Total 42 cases were studied, and the age of patient ranged from 33-80 years. Out of 42 cases of IDC-NST (Invasive ductal carcinoma- No special type) 19 cases (45%) showed positive expression and 23 cases (54%) showed lack of staining for Bcl-2. Tumor size and grade of IDC-NST has significant relationship with Bcl-2 expression with p value of 0.05. No correlation has been found between lymph nodes harboring malignant cells and Bcl2 expression with p value &gt;0.05. Conclusion: The staging and prognosis of breast cancer depend heavily on the size, grading based on histopathology, and lymph node harboring tumor cells. Hormonal receptor tests are frequently used in the process of treating breast cancer. The goal of the current investigation was to assess Bcl2's value as an additional marker for prognosis. Significant Bcl-2 expression was seen in tumors of modest histologic grade and size.

Inflammatory and proapoptotic effects of inhaling gasoline fumes on the lung and ameliorative effects of fenugreek seeds

Impacts of inhaling gasoline fumes on the lungs of adult male rats and the alleviating role of fenugreek seeds were evaluated. Twenty-four rats were divided into four groups, unexposed control and fenugreek groups, gasoline exposed groups for 6 h/6 day/week for 10 weeks with and without supplementation of fenugreek seed powder in food (5% w/w). Rats exposed to gasoline fumes showed significant elevation in lung tumor necrosis factor-α, as an inflammatory marker, and the proapoptotic marker Bax with a reduction in the antiapoptotic marker Bcl2. Moreover, remarkable elevations in transforming growth factor-β1, collagen and hydroxyproline were observed as fibrotic markers. Lung oxidative stress markers (hydrogen peroxides, malondialdehyde, and protein carbonyl) increased significantly along with marked decrease in total antioxidant capacity, superoxide dismutase, and catalase levels. Additionally, marked decreases in white and red blood cell counts, hemoglobin content, platelet count, accompanied by elevated red cell distribution width percentage were observed, supporting the inflammatory status. Histopathological changes represented by hematoxylin&eosin, immunohistochemistry staining for Bax&Bcl2, and transmission electron microscopy supported the negative impacts of gasoline fumes compared to the control group. Fenugreek seeds supplementation with gasoline exposure showed pronounced alleviation of lung biochemical and histopathological changes compared to the gasoline-exposed group.

Focal Isolated Extraocular Muscle Lymphoma

A 61-year-old woman presented with a painless left subconjunctival nodule (Fig A). Magnetic resonance imaging revealed a heterogeneously enhancing mass at the insertion of the left medial rectus muscle on postcontrast T1-weighted images (Fig B). Incisional biopsy was performed. Histopathology demonstrated fibrous tissue with atypical lymphoid infiltrate (Fig C, Hematoxylin and eosin stain) and positive CD20 (Fig D) and BCL-2 staining consistent with extranodal marginal zone lymphoma (EMZL). The patient was treated with external beam radiotherapy with complete remission.

The effect of tropisetron on peripheral diabetic neuropathy: possible protective actions against inflammation and apoptosis.

Diabetic peripheral neuropathy (DPN) is a common nerve disorder of diabetes. The aim of this study was to explore the protective effects of tropisetron in DPN. Type 1 diabetes was created by a single injection of streptozotocin (50 mg/kg, ip). Tropisetron (3 mg/kg, ip) was administered daily for 2 weeks. Our analysis showed that nerve fibers and their myelin sheaths were thinned with decreased myelinated fiber number in diabetic animals. The intensity of Bcl-2 staining decreased and the intensity of Bax staining increased in the sciatic nerves of diabetic rats by using immunohistochemical staining. Furthermore, diabetes significantly increased tumor necrosis factor-alpha, interleukin 1-β (TNFα and IL-1β) and Bax/Bcl-2 ratio in sciatic nerves of rats. However, intraperitoneal injection of tropisetron significantly reversed these alterations induced by diabetes. These findings suggest that tropisetron attenuates diabetes-induced peripheral nerve injury through its anti-inflammatory and anti-apoptotic effects, and may provide a novel therapeutic strategy to ameliorate the process of peripheral neuropathy in diabetes.

Rapid death due to pulmonary epithelioid haemangioendothelioma in several weeks: A case report

A 49-year-old woman was admitted to our hospital because of haemoptysis for 6 days. This patient claimed no medical history except high blood sugar. Chest computed tomography (CT) showed infection and multiple nodules on both sides of the lung. Blood tests showed no obvious abnormalities. Tracheoscopy showed haemorrhagic discharge in the left upper lobe and an old thrombus obstructing the lumen in the anterior basal segment of the right lower lobe. Then, CT-guided percutaneous lung biopsy was performed. The pathological results suggested multiple nodular-like lesions in the submitted tissues, and tumour cells were round or short fusiform, forming a solid nest structure, visible mitosis, and a vascular cavity-like structure containing red blood cells. Immunohistochemistry revealed positive staining for Vimentin, Bcl-2, CD31, and CD34; negative staining for CD68, SMA, CR, and D2-40; and 40% Ki67+ positivity. Based on the earlier data, the patient was diagnosed with pulmonary epithelioid haemangioendothelioma. This patient did not receive any treatment for several reasons. Unfortunately, the patient died 8 weeks after diagnosis. In conclusion, we present a case featuring the rapid death due to PEH.

The neuroprotective effects of transcranial direct current stimulation on global cerebral ischemia and reperfusion via modulating apoptotic pathways

BackgroundCerebral ischemia-reperfusion, subsequent hyperthermia, and hyperglycemia lead to neural damage. This study aimed to investigate the effects of using cathodal and/or anodal transcranial direct current stimulation (tDCS) in different stages of ischemia-reperfusion on apoptosis and controlling hyperthermia and hyperglycemia. Materials and MethodsA total of 78 male Wistar rats were randomly assigned into six groups (n = 13), including sham, ischemia/reperfusion (I/R), anodal-tDCS (a-tDCS), cathodal-tDCS (c-tDCS), anodal/cathodal-tDCS (a/c-tDCS), and cathodal/anodal-tDCS (c/a-tDCS) groups. Global cerebral I/R was induced in all of the groups except for sham group. In a-tDCS and c-tDCS groups, the rats received anodal and cathodal currents in both I/R stages, respectively. In a/c-tDCS group, the rats received anodal current during the ischemia and cathodal current during the reperfusion. The c/a-tDCS group received the currents in the reverse order. The current intensity of 400 µA was applied in ischemia phase (15 min) and reperfusion phase (30 min, twice a day). Body temperature and plasma blood sugar were measured daily. Rats were also tested for novel object recognition and passive avoidance memory. The apoptosis of hippocampal tissue was evaluated by measuring Bax, Bcl-2, Caspase-3, and TUNEL staining. ResultsAll tDCS significantly reduced hyperthermia and hyperglycemia, as well as Bax and Caspase-3 levels, it also increased Bcl-2 expression. The preliminary results from c/a-tDCS mode could improve the expression of apoptotic markers, memory function, hyperthermia, and hyperglycemia control and reduce DNA fragmentation compared to other stimulatory therapies. ConclusionAll tDCS modes could save neurons by suppressing apoptotic and enhancing anti-apoptotic pathways, especially in the c/a tDCS mode.

Developmental Characteristics of the Glomerular and Tubular Portion of the Nephron in the Human Foetal Kidney Cortex: Morphometrical and Immunohistochemical Analysis

This study aimed to morphometrically examine the development of glomeruli and tubules in the kidney cortex of human foetuses at different gestational ages (GAs). We also investigated the expression of the proliferation marker Ki-67 and apoptosis-related markers Bcl-2 and Bax during nephrogenesis using immunohistochemistry. Kidney samples from 38 human foetuses of both sexes with GA ranging from 13 to 40 weeks were analysed. The samples were divided into 7 groups based on GA, each corresponding to 1 lunar month. Foetal kidneys showed a spatiotemporal gradient of nephron differentiation with the transient stages of nephron anlage located in the nephrogenic zone and immature nephrons located in the subjacent maturation zone. In the inner cortex, nephrons establish the morphological characteristics of definitive nephrons. The average area, perimeter, and Feret’s diameter of the glomeruli formed within the kidney cortex gradually decreased up to a period of 29–32 weeks of gestation and subsequently increased until a period of 37–40 weeks. There was a weak negative correlation with GA. In contrast, the areal density of glomeruli increased up to a period of 21–24 weeks and then gradually decreased until a period of 37–40 weeks, showing a moderate negative correlation with GA. The average area of renal tubules slightly decreased until a period of 21–24 weeks of gestation and then gradually increased until a period of 36–40 weeks, showing a moderate positive correlation with GA. The average areal density of renal tubules increased significantly until a period of 21–24 weeks of gestation, remained relatively constant until a period of 33–36 weeks, and then increased significantly at 36–40 weeks. There was a strong positive correlation with GA. Our results showed that Ki-67 was expressed in numerous cells of the metanephric mesenchyme, pretubular aggregates, renal vesicles, comma-shaped bodies, and early S-shaped bodies. During subsequent development and the spatial expansion of nephrons towards the mature zone, the expression of Ki-67 was markedly reduced. Similarly, Bcl-2 was strongly expressed in induced nephrogenic progenitor cells, pretubular aggregates, renal vesicles, and comma-shaped bodies. As vascularisation and maturation of the nephron proceeded, Bcl-2 staining became less intense and limited to the parietal layer of the Bowman’s capsule and renal tubules. Weak Bax expression was observed in individual scattered cells within segments of the nephrons at all developmental stages. In the mature zone, more intense Bax staining was observed in the renal tubules.

Anti-Cancer Effect of Phosphatidylcholine Containing Conjugated Linoleic Acid at sn-2 Position on MCF-7 Breast Cancer Cell Line

Conjugated linoleic acid (CLA) exhibits anti-cancer effects; however, most studies have only investigated its free fatty acid forms (CLA-FFA). Here, we compared the anticancer effect of CLA with its lipid forms (FFA and phosphatidylcholine [PC]) on the breast cancer cell line MCF-7 with Soy-PC as a control. CLA-FFA, CLA-PC, and Soy-PC caused a dose- and time-dependent inhibition of MCF-7 growth. However, the lowest antiproliferative activity was observed with CLA-PC. These results were consistent with Hoechst staining and protein expression of apoptosis markers Bax and Bcl-2. Interestingly, CLA-PC uptake was four times lower than that of Soy-PC, which may have affected its antiproliferative effect. Many papers showing that phospholipid forms of functional compounds have superior activity than intact forms have been reported, however, our results suggest that the PC form of CLA was inferior to its FFA form in inhibiting MCF-7 cell growth. This is the first report showing that the phospholipid form is not always the superior form.

Meprin β mediates Osteopontin‐Signaling in IR‐Induced Acute Kidney Injury

Ischemia/reperfusion (IR) causes acute kidney injury (AKI) and is associated with high morbidity and mortality rates. Meprins metalloproteases are expressed abundantly in the brush border membranes of proximal kidney tubules and implicated in the progression of AKI. However, the underlying mechanisms are not fully understood. Meprins are capable to proteolytically processing osteopontin (OPN), a highly phosphorylated multifunctional and matricellular glycophosphoprotein. OPN mediates cell signaling pathways involved in apoptosis and extracellular matrix metabolism. The objective of the present study was to determine the effect of meprin β expression on OPN and downstream mediators of the OPN‐signaling pathway in IR‐induced renal injury. Surgical procedures were used to induce unilateral IR in wild‐type (WT) and meprin β knockout (βKO) mice. The mice were sacrificed at 24h post‐IR. Blood samples and kidney tissues were obtained for analysis. Neutrophil gelatinase associated lipocalin (NGAL) was used for biochemical assessment of kidney injury. Real‐time PCR, immunohistochemical staining, and western blot analysis were used to evaluate the levels of OPN, caspase‐3, Bcl‐2, and NFκB. Statistical analysis utilized 2‐way ANOVA. Our data show that NGAL levels were increased significantly in the kidney of WT (P=0.0001) and βKO (P=0.01) mice at 24h post‐IR when compared to their control counterparts. OPN mRNA increased in both genotypes at 24h post‐IR. Western blot analysis detected a full length OPN protein band at 66 kDa, with significant increases (p=0.001) in protein levels in WT kidney at 24h post‐IR when compared to levels in control kidneys. This increase in OPN was not observed in meprin βKO mice subjected to IR. Additionally, we detected a cleaved OPN fragment at 32 kDa, for which there were no significant changes in either genotype at 24h IR post‐IR. Immunohistochemical staining showed the increase in the levels of OPN occurred in select WT kidney tubules and in renal corpuscles. To determine whether meprin β expression correlates with the increased OPN, we used immunofluorescence counterstaining with proximal tubule (PT) markers (meprin β for WT and villin for βKO). Our data show that the increase in OPN occurred in both proximal and distal kidney tubules. Interestingly, higher levels of OPN were detected in the lumen of PTs from WT but not βKO kidneys, suggesting that meprin β is partly responsible for enhanced release of OPN into filtrate and ultimately into urine as previously reported. OPN functions as anti‐apoptotic protein in different diseases, therefore we evaluated its impact on caspase‐3, Bcl‐2, and NFκB. The mRNA levels for caspase‐3, Bcl‐2 and NFκB decreased in WT but increased in βKO mice at 24h post‐IR. Western blot detected higher levels of caspase‐3 and Bcl‐2 only in WT kidneys. Immunohistochemical staining for caspase‐3, Bcl‐2 and NFκB also showed higher levels in tubules and renal corpuscle of WT kidneys when compared to βKO kidneys at 24h post‐IR. Taken together, our data show that meprin β regulates OPN and down‐stream mediators of OPN‐signaling pathway in IR‐induced AKI.

Oral Lipomatous Neurofibroma: A report of two cases and a review of the literature

Neurofibroma is a benign tumor that originates from the peripheral nerve sheath. This lesion may assume multiple growth patterns and various histologic variants have been described in the literature. We present two cases of lipomatous neurofibroma, a rare variant in the oral cavity. This variant is typically seen as a cutaneous lesion predominately located in the head and neck. Case 1 was a 62-year-old female who presented with a mass of her left posterior palate. Case 2 presented as a submucosal dorsal tongue mass in an 18-year-old male. Both lesions appeared clinically benign. Neither patient had a known history of neurofibromatosis type 1. The biopsies from case 1 and case 2 showed similar histopathological features. These two lesions were well delineated submucosal tumors characterized by an admixture of spindled neural cells and mature adipocytes. The mature adipocytes were separate from the regional adipose tissue. The spindle cells contained serpentine nuclei set in a wavy well-vascularized fibrillar stroma. The immunohistochemical staining for both lesions revealed S100 positivity of the tumor cells and negative staining for CD99, CD34, and BCL2. Differential diagnosis for these tumors included solitary fibrous tumor and spindle cell lipoma. However, based on the lack of a hemangiopericytoma-like vascular pattern, wire-like collagen and the pattern of IHC staining, these entities were excluded. While the diagnosis of a conventional neurofibroma is straightforward, the diagnosis of a lipomatous neurofibroma may be more challenging. Neither of our patients had a known history of Neurofibromatosis type 1 (NF1); however, cutaneous lipomatous neurofibromas have been associated with NF1. Therefore, a patient with a diagnosis of lipomatous neurofibroma should have a medical work-up to exclude this syndrome. With the presentation of two cases of lipomatous neurofibroma arising intraorally, we hope to raise awareness of this distinct and rare morphologic subtype of neurofibroma.

Comparison of melatonin, oxytetracycline, and N-acetylcysteine pre-treatments in autologous intraperitoneal ovarian transplantation in rats

This study was aimed at investigating the effects of melatonin, oxytetracycline and N-acetylcysteine on the ovarian follicle reserves and surface epithelium in autologous intraperitoneal ovarian transplantation in rats. Thirty adult female Wistar Albino were selected and randomly divided into six groups (n = 5). Group 1, which was the control group, only had their abdomens opened and closed while Group 2 underwent ovarian transplantation. Group 3, 4, 5 and 6 received 20 μg/kg/IM melatonin, 10 mg/kg/IM oxytetracycline, 150 mg/kg/IP N-Asetil sistein (NAC) and 1% ethanol respectively 15 min before the ovarian transplantation. Vaginal cytology was performed to monitor the estrus phase and the follicle reserve and changes in the surface epithelium were histopathologically evaluated during the preparations. Moreover, cellular apoptosis in tissues was evaluated with immunofluorescence staining of Bcl-2 and Bax. The Bax/Bcl-2 ratio was then calculated as the mean fluorescence intensity (MFI) of Bax and Bcl-2 MFI. Dysplastic change was found only significantly higher in the transplantation group (G2) (p < 0.01). Histopathologically, it was found that the follicle reserve was preserved significantly in the oxytetracycline and melatonin treated group (G3, G4) (p < 0.01). It was also observed that the oxytetracycline treated group (G4) were able to show better preventive effects against dysplastic changes of the surface epithelium. Moreover, the melatonin treated group depicted a low Bax/Bcl-2 ratio compared to the group that only underwent transplantation (G2) (p < 0.01). This study indicated that oxytetracycline and melatonin might be more effective than N-acetylcysteine in protecting against oxidative stress during ovarian transplantation.

Corn embryo ameliorates cognitive dysfunction and anxiety-like behaviors in D-galactose-induced aging rats via attenuating oxidative stress, apoptosis and up-regulating neurotrophic factors

Aging is the primary cause of neurodegenerative diseases, which are mainly characterized by cognitive decline and neuropsychiatric symptoms. Corn embryo, an important component of corn kernels, contains plenty of essential nutrients and bioactive compounds. However, corn embryo is often removed in the process of refining corn. To reveal potential biological benefits of corn embryo, the present study investigated the intervention effects of corn embryo on age-related cognitive decline and neuropsychiatric symptoms. Ninety male Wistar rats were randomly divided into six groups: Control, Corn embryo, Aging model, Low-, Medium- and High-dose intervention group. Aging models induced by an intraperitoneal injection of 60 mg/kg D-galactose plus a gavage of 200 mg/kg aluminum chloride were intervened with a gavage of 0.3, 0.6 or 1 g/kg corn embryo while the Control and Corn embryo groups received saline and 0.6 g/kg corn embryo respectively. Morris water maze and open field test were performed to assess cognitive abilities and anxiety-like behaviors. Brain biochemical parameters including the malondialdehyde, glutathione, glutathione sulfhydryl transferase and γ-glutamylcysteine synthetase were detected to evaluate oxidative stress levels. The mRNA expression of brain-derived neurotrophic factor was determined to estimate neurotrophic factor levels. Besides, histopathological alterations were visualized by hematoxylin-eosin staining and neuronal apoptosis levels were measured by the immunohistochemical staining of Bax and Bcl-2. Ultimately, the mimetic aging rats showed significant cognitive impairment (n = 15, P < 0.01) and anxiety-like behaviors (n = 15, P < 0.01), increased oxidative stress (n = 5, P < 0.001), neurodegeneration (n = 5, P < 0.001) and apoptosis (n = 5, P < 0.01) and reduced neurotrophic factors (n = 5, P = 0.074) in the brain. However, corn embryo effectively prevented the above undesirable neurobehavioral alterations via attenuating oxidative stress (n = 5, P < 0.01), neurodegeneration (n = 5, P < 0.001) and apoptosis (n = 5, P < 0.01) and increasing the levels of neurotrophic factors (n = 5, P < 0.001), suggesting its strong neuroprotective effects.

English

BACKGROUND Bcl-2 gene is an apoptotic protein that blocks apoptosis and thereby its over expression contributes to neoplastic transformation and decreased tumour survival. So, it is necessary to find out the relationship of Bcl-2 expression with histological types and tumour grade in ovarian surface epithelial tumours, which may predict the prognosis. METHODS The objective was to study the expression of Bcl-2 in ovarian surface epithelial tumours and to correlate Bcl-2 expression with histopathological features and tumour grade in ovarian surface epithelial tumours. Histological types and tumour differentiation for each case is determined from the routine H and E sections. Immunohistochemical stain for Bcl-2 was done. Then intensity and extent of staining for Bcl-2 was compared with the age, histological type and tumour grade. RESULTS Out of the 47 cases studied, 66% were in &lt;55 years of age category and 34% in &gt;55 years of age group. There was statistically significant associations of Bcl-2 expression with various histological types (P&lt;0.001) and tumour differentiation (P&lt;0.001). In the case of extent of Bcl-2 staining, statistically significant associations were present with various histological types (P=0.004) and tumour differentiation (P&lt;0.001). CONCLUSIONS Bcl-2 expression decreases with tumour progression. Poorly differentiated tumours with decreased Bcl-2 expression may be helpful in predicting disease progression. Further studies are warranted since, Bcl-2 expression may be important for prognostic outcome or provide useful targets for therapeutic intervention in patients with surface epithelial ovarian cancers. KEY WORDS Bcl-2, Immunohistochemistry, Tumour Grade, Epithelial Tumours.

Dual targeting nanoparticles based on hyaluronic and folic acids as a promising delivery system of the encapsulated 4-Methylumbelliferone (4-MU) against invasiveness of lung cancer in vivo and in vitro

Lung cancer is the most common cause of cancer death worldwide. Thereby, new treatment strategies as targeting nano-therapy present promising possibilities to control the aggressiveness of lung cancer. Dual CD44 and folate receptors targetable nanocapsule based on folic-polyethylene glycol-hyaluronic (FA-PEG-HA) were fabricated to improve the therapeutic activity of 4-Methylumbelliferone (4-MU) toward lung cancer. In this study, we fabricate 4-MU Nps as a hybrid polymeric (protamine) protein (albumin) nanocapsule, then functionalized by targeting layer to form 4-MU@FA-PEG-HA Nps with encapsulation efficacy 96.15%. The in vitro study of free 4-MU, 4-MU Nps and 4-MU@FA-PEG-HA Nps on A549 lung cancer cells reveal that the 4-MU Nps and 4-MU@FA-PEG-HA Nps were more cytotoxic than free 4-MU on A549 cells. The observed therapeutic activity of 4-MU@FA-PEG-HA Nps on urethane-induced lung cancer model, potentiality revealed a tumor growth inhibition via apoptotic mechanisms and angiogenesis inhibition. The results were supported by Enzyme-linked immunosorbent assay (ELIZA) of transforming growth factors (TGFβ1) and serum HA, histopathological analysis as well as immunohistochemical Ki67, CD44, Bcl-2 and caspace-3 staining. Moreover, 4-MU@FA-PEG-HA Nps exhibited a promising safety profile. Hence, it is expected that our developed novel nano-system can be used for potential application on tumor therapy for lung cancer.

A small round blue cell tumour of the rib: A case report of Ewing sarcoma

Ewing sarcoma is a prototypical small round blue cell tumour with a characteristic EWSR1 translocation. We report a case of Ewing sarcoma arising in the rib of an 81-year-old female. The tumour was comprised of uniform small round cells which demonstrated an organoid growth pattern with a suggestion of rosette-like structures. Immunohistochemistry showed positive staining for CD99, vimentin and desmin; stains for cytokeratins, melanocytic markers, neuroendocrine markers, CD45, CD138, NFP and CD34 were negative. EMA and BCL2 demonstrated weak, patchy staining. FISH studies demonstrated an EWSR1 fusion, supporting the diagnosis of Ewing sarcoma. Desmin positivity, although recognised in Ewing sarcoma, has been reported in only 2% of cases.1 The main differential diagnosis based on morphology and immunohistochemistry was between Ewing sarcoma and synovial sarcoma due to CD99 positivity and focal EMA and BCL2 staining. However, EWSR1 translocations are not associated with synovial sarcoma.2 Recently a proportion of ‘undifferentiated round cell sarcomas’ with similar morphology to Ewing sarcoma have been divided into specific entities largely based on their molecular characteristics. These include CIC-rearranged sarcomas, sarcomas with BCOR alterations and round cell sarcomas with EWSR1-non ETS fusions.3

Immunoexpression of REGγ and apoptosis-related proteins in oral tongue squamous cell carcinoma

The aim of this study was to investigate the immunohistochemical expression of REGγ, p53, MDM-2, Bcl-2, and Bax in oral tongue squamous cell carcinoma (OTSCC), and to correlate the findings with clinicopathological parameters. Fifty-eight OTSCC cases were selected for the study. The percentages of nuclear (REGγ, p53, and MDM-2) and cytoplasmic (Bcl-2 and Bax) staining in epithelial cells were determined and correlated with clinicopathological parameters (regional lymph node metastasis, clinical stage, clinical outcome, and histopathological grade of malignancy). Expression of REGγ was observed in all cases studied. Significantly lower percentages were observed in tumours with lymph node metastasis (P = 0.036) and in high-grade tumours (P = 0.013). No significant differences in p53, MDM-2, or Bax expression were observed according to the clinicopathological parameters. Lower percentages of Bcl-2 staining were found in high-grade OTSCC (P = 0.040) and in cases of disease-related death (P = 0.032). The expression of REGγ showed a weak positive correlation with the expression of MDM-2 (P = 0.001) and Bcl-2 (P = 0.014). The results of this study suggest that lower expression of REGγ may contribute to the progression of OTSCC. The role of REGγ in the development of OTSCC does not appear to be primarily related to the modulation of apoptosis in neoplastic cells.