Difference In Bcl-2 Expression Research Articles

Exploring the Role of EZH2 and BCL2 in Demarcating Oral Verrucous Hyperplasia and Verrucous Carcinoma

Introduction Oral verrucous hyperplasia (OVH) and verrucous carcinoma (OVC) are precursors of oral squamous cell carcinoma exhibiting overlapping histopathological picture which warrants distinction. EZH2 is an epigenetic marker possessing multifaceted function in cellular proliferation, migration, and malignant transformation, whereas BCL2 is an integral part of the antiapoptotic mechanism regulating cellular homeostasis. Aim The aim was to distinguish OVH and OVC by analysis of immunohistochemical expression of EZH2 and BCL2. Material and Methods The study sample consisted of 79 formalin-fixed paraffin-embedded tissue sections of normal oral mucosa (10), OVH (10), oral OVC (27), and oral squamous cell carcinoma (32). Immunohistochemical analysis of EZH2 and BCL2 was done and labeling indices were calculated. Additionally, six histopathological parameters were assessed in OVH and OVC. Statistical analysis was done using Kruskal–Wallis test, Tukey honest significant difference test, and Spearman's correlation. Receiver operating characteristic curve was plotted and sensitivity, specificity, and cutoff score of each marker were calculated. Result and Discussion Labeling indices of EZH2 and BCL2 depicted a gradual incline from normal mucosa to oral squamous cell carcinoma. Significant difference of EZH2 and nonsignificant difference in BCL2 expression between OVH and OVC were noted. Out of the six histopathological parameters, keratin plugging, juxtaepithelial lymphocytic response, and frank endophytic growth yielded a significant difference. EZH2 serves as a superior marker than BCL2 to differentiate OVH and OVC. Juxtaepithelial lymphocytic response can also serve as a histopathological parameter in distinguishing OVH and OVC.

Biological differences and molecular mechanism of different peripheral nerves after injury.

The aim of the study was to analyze the apoptosis of neurons and the differences in expression of Bcl-2 and Bax protein in the neurons in the corresponding spinal cord segment after the repair of the tibial nerve (TN) and common peroneal nerve (CPN) in rats. 126 healthy male Sprague-Dawley (SD) rats aged 7-8 weeks were randomly divided into group A (control group), group B (TN was cut and sutured), and group C (CPN was cut and sutured), with 42 rats in each group. The spinal cord tissues of rats in different groups were stained with hematoxylin-eosin (HE) on the 1st, 3rd, 7th, 14th, 21st, and 28th day after surgery; the number of neurons in anterior horn of spinal cord, axon density (AD), axon passage rate (APR), and recovery rate (RR) of muscle cell cross-sectional area (MCCA) were calculated; and differences in the expression of Bcl-2 and Bax proteins in the three groups of rats were analyzed by immunohistochemistry. The results showed that there was no statistically significant difference in the muscle wet weight (MWW) RR of the three groups of rats on the 14th day after the surgery (p>0.05), and the MWW RRs of rats in groups B and C were higher at the 28th day after surgery in contrast to group A (p<0.05). The number of motor neurons in the anterior horn of spinal cord in group B was higher than that in group C at the 3rd, 7th, 14th, and 21st day after surgery (p<0.05); the MWW RR, MCCA, and CSARR of rats in group B were lower than those in group C (p<0.05); the proximal AD, distal AD, and APR in group B were higher than those of group C on the 14th and 28th day after the surgery (p<0.05); and there were no positive staining results in the spinal cord tissue of rats in group A after staining. The expressions of Bcl-2 and Bax in group B were higher observably than the expressions in group C (p<0.05), which indicated that the recovery ability of TN was stronger than that of the CPN; the expression of Bcl-2 and Bax in TN was notably higher than that of the CPN. The expression of Bcl-2 and Bax was related to cell apoptosis and nerve regeneration after nerve injury. It provided a reference basis for clinical diagnosis and treatment of peripheral nerves.

Phoenix dactylifera L. Pollen and Fluvoxamine Maleate Protect PC12 Cells Against H2O2-Induced Oxidative Stress by Involvement of Nrf2 and SIGMAR1 Gene Expression

Background: The association between oxidative stress and the pathogenesis of neurodegenerative diseases has been documented. Besides, there is evidence that antidepressant agents, such as fluvoxamine maleate (Flv), can ameliorate neurotoxicity. The date palm (Phoenix dactylifera L.) pollen (DPP) contains various compounds with antioxidant capacity; however, its underlying mechanism of function has not been fully understood. Objectives: The present study aimed to compare the neuroprotective effects of DPP and Flv on H2O2-induced oxidative damage and their effects on Nrf2, SIGMAR1, and Bcl2 gene expression in PC12 cells. Methods: First, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay examined the toxicities of DPP, Flv, and H2O2 at various concentrations on the PC12 cells. Real-time polymerase chain reaction (PCR) measured the expression of Nrf2, SIGMAR1, and Bcl2 in PC12 cells in the presence or absence of DPP or Flv after 4 h of treatment with 100 μM H2O2. Results: Based on the MTT results, DPP at concentrations of 200 - 1000 µg/mL had no toxic effect on PC12 cells. The IC50 was evaluated at 57.26 μM and 109.5 μM under treatment with Flv and H2O2. Real-time PCR analysis showed a decrease in the expression of Nrf2 and SIGMAR1 in PC12 cells treated with 100 μM H2O2, an indicator of oxidative stress recruitment in PC12 cells. Pretreatment with DPP and Flv (500 µg/mL and 10 µM, respectively) for 24 h resulted in the upregulation of Nrf2 relative to the vehicle control. In addition, pretreatment with DPP increased the level of SIGMAR1 in PC12 cells compared with H2O2-exposed cells. Considering the role of SIGMAR1 in endoplasmic reticulum oxidative stress, the SIGMAR1 level should be evaluated at the translational level. Compared to the untreated cells, the expression of Bcl2 decreased in all the experimental cases, and the difference in Bcl2 expression was not significant between the co-treatment experimental cases. Conclusions: Taken together, DPP and Flv have neuroprotective effects against oxidative damage in PC12 cells. Exposure of PC12 cells to 500 μg/mL DPP and 10 μM Flv for 24 h protected the morphology and function of PC12 cells under H2O-induced oxidative stress via regulating oxidative stress-involved genes.

Sex‐differences and cell type‐specific alterations in brain Bcl2 expression after transient focal cerebral ischemia in aged mice

IntroductionStroke remains the second leading cause of death worldwide and remains the primary cause of long‐term disability in the US. To date, the only pharmacological treatment for stroke remains tissue plasminogen activator. A major factor for translational failure is likely overwhelming use of young male rodents despite the fact that stroke occurs predominantly in aged populations and is a sexually dimorphic disease with sex differences in incidence, prevalence, and outcomes. The Bcl2 family is known to play an important role in the evolution of injury following cerebral ischemia. Overexpression of pro‐survival Bcl2 protects against cerebral ischemia in vivo and in vitro. In the present study we explored whether Bcl2 could play a role in known age‐ and sex‐related differences in stroke.MethodsAll studies were conducted in accordance with National Institutes of Health guidelines for the use of experimental animals, with protocols approved by the Stanford Animal Care and Use Committee. Transient focal cerebral ischemia was induced by suture occlusion of the middle cerebral artery (MCAO) for 1h in 20 month‐old male and female C57BL/6 mice. Sham surgery consisted of anesthesia and internal carotid exposure without suture occlusion (n=6 per group). Histological assessment: Animals were sacrificed at post‐MCAO days 1, 3, and 30 (n=6 per group, per time point), and brains fixed with 4% phosphate‐buffered paraformaldehyde for stereological analysis. Fixed sections were stained for Bcl2, the astrocyte marker glial fibrillary acidic protein (GFAP), and the mature neuronal marker NeuN. An observer blinded to conditions quantified the cell‐type specific relative intensity of Bcl2 from maximum projection Z‐stack images.ResultsIn sham animals Bcl2 expression was significantly (p&lt;0.05) higher in both female astrocytes and neurons relative to male. After MCAO male neuronal Bcl2 expression was significantly depressed from days 1 until 30, while female neuronal Bcl2 remained unchanged. Conversely in astrocytes, female Bcl2 expression decreased significantly at post‐injury days 1 and 3, but recovered by day 30, while male astrocyte Bcl2 decreased by day 3 and remained significantly repressed at day 30.ConclusionsBaseline differences in Bcl2 expression between aged males and females, and cell‐type specific differences in post‐injury Bcl2 expression may account for age‐ and sex‐related differences in injury outcome from stroke. Bcl2 may represent a therapeutic target for stroke for both sexes in clinically‐relevant aged populations.

Propofol-induced sleep ameliorates cognition impairment in sleep-deprived rats.

Propofol has been shown to clear sleep debt in rats after sleep deprivation (SD). We examined whether or not propofol-assisted sleep can restore cognitive function in SD rats and explored the possible mechanisms. A sleep deprivation model was established by housing 9 to 12 week-old rats to a multiplatform water tank for 96h. Model rats were then intraperitoneally injected with different concentrations of propofol or 10% fat emulsion (vehicle control). All treatment groups were examined for spatial learning and memory ability in the Morris water maze (MWM). After euthanasia, morphological changes in the hippocampus, hippocampal neurons, and mitochondria were examined by hematoxylin-eosin staining and transmission electron microscopy. Serum and hippocampal levels of IL-1β, TNF-α, and hippocampal concentrations of ATP and Cyt-c were measured by ELISA (enzyme-linked immunosorbent assay). Immunohistochemistry and Western blotting were performed to assess hippocampal expression of Bcl-2, Bax, and cleaved caspase-3. Results showed that escape latencies in MWM training trials were significantly shorter and target crossings in the memory probe trial significantly greater in propofol-treated SD model rats compared to vehicle-treated SD rats. Propofol also reduced the number of apoptotic bodies in the hippocampal CA1 region. Sleep deprivation reduced IL-1β and ATP in hippocampus while increasing TNF-α and Cyt-c, and propofol treatment reversed all these changes. There was no significant difference in Bcl-2 expression between propofol- and vehicle-treated SD rats, but pro-apoptotic Bax and cleaved caspase-3 expression levels were significantly reduced by propofol in SD rats. Propofol-assisted sleep restored cognitive function in SD rats possibly by attenuating mitochondria-mediated neuronal apoptosis in the hippocampus.

Naïve CD4+ T Cell Lymphopenia and Apoptosis in Chronic Hepatitis C Virus Infection Is Driven by the CD31+ Subset and Is Partially Normalized in Direct-Acting Antiviral Treated Persons.

BackgroundThe mechanisms underlying naïve CD4+ lymphopenia during chronic Hepatitis C Virus (HCV) infection are unclear. Whether direct-acting antiviral (DAA) therapy restores peripheral naïve CD4+ T cell numbers and function is unknown.MethodsWe enumerated frequencies and counts of peripheral naïve CD4+, CD4+CD31+ and CD4+CD31- T cells by flow cytometry in a cross sectional analysis comparing chronic HCV infected (n=34), DAA-treated(n=29), and age-range matched controls (n=25), as well as in a longitudinal cohort of HCV DAA treated persons (n=16). The cross-sectional cohort was stratified by cirrhosis state. Cell apoptosis/survival (AnnexinV+7AAD+/BCL-2 labeling) and cell cycle entry (Ki67 expression) of CD31+ and CD31- naïve CD4+ T cells was analyzed directly ex vivo and following 3 and 5 days of in vitro culture with media, interleukin (IL) -7 or CD3/CD28 activator.ResultsIn the cross-sectional cohort, naïve CD4+ proportions were lower in chronic HCV infected persons compared to controls and DAA-treated persons, an effect in part attributed to cirrhosis. Age was associated with naïve cell counts and proportions in HCV infected and treated persons as well. Naïve CD4+ cell proportions negatively correlated with plasma levels of soluble CD14 following therapy in DAA-treated persons. Naïve CD4+ cells from HCV infected persons exhibited greater direct ex vivo apoptosis and cell-cycling compared to cells from DAA-treated persons and controls, and this was localized to the CD4+CD31+ subset. On the other hand, no remarkable differences in expression of BCL-2 or IL-7 Receptor (CD127) at baseline or following in vitro media or IL7 containing culture were observed. In the longitudinal cohort, naïve CD4+CD31+/CD31- ratio tended to increase 24 weeks after DAA therapy initiation.ConclusionsActivation and apoptosis of peripheral naïve CD4+CD31+ T cells appear to contribute to naïve CD4+ lymphopenia in chronic HCV infection, and this defect is partially reversible with HCV DAA therapy. Age and cirrhosis -associated naïve CD4+ lymphopenia is present both before and after HCV DAA therapy. These findings have implications for restoration of host immune function after DAA therapy.

CD34 and Bcl-2 as predictors for the efficacy of neoadjuvant chemotherapy in cervical cancer

BackgroundSuccessful neoadjuvant chemotherapy (NACT) could improve the surgical resection rate and radical curability of patients with cervical cancer, but only a subset of patients benefits. Therefore, identifying predictive biomarkers are urgently needed. The aim of this study was to evaluate the predictive value of CD34 and Bcl-2 in the NACT effectiveness of cervical cancer.MethodsSixty-seven patients with locally advanced cervical cancer (FIGO stages IB3, IIA2 or IIB) were classified into two groups based on effective (n = 48) and ineffective (n = 19) response to NACT. Immunohistochemistry was employed to identify CD34 and Bcl-2 expression before and after NACT. We analyzed the associations between the pre-NACT expression of these two biomarkers and the response of NACT. The expression of these two biomarkers before and after NACT was also assessed and compared.ResultsMore patients were CD34 positive expression before NACT in effective group compared to ineffective group (p = 0.005). However, no statistically significant difference in Bcl-2 expression before NACT were found between two groups (p = 0.084). In NACT effective group, the expression of both CD34 and Bcl-2 after NACT are down-regulated (p < 0.001 and p < 0.001, respectively), while there are no statistical differences between the pre- and post-NACT expression of CD34 and Bcl-2 in NACT ineffective group (p = 0.453 and p = 0.317, respectively).ConclusionThe positive CD34 expression before NACT may serve as a predictive biomarker for NACT of cervical cancer, but the pre-NACT expression of Bcl-2 is not an independent predictor. The down-regulated expression of these two indicators after NACT may indicate effective NACT.

IgM-Secreting Diffuse Large B-Cell Lymphoma (DLBCL) Is a Poor Prognostic Subset within the Non-Germinal-Centre-Type (GC-type): An Italian Multicentre Study

BACKGROUND: In 2014 we identified a new subset of DLBCL, defined as "IgM-secreting" (Cox MC &amp; Di Napoli A , PLOS One 2014). This was characterised by poor prognostic features and outcome as well as frequent central nervous (CNS) system localizations. Furthermore, IgM-secretion, was an independent prognostic factor in multivariate analysis. Here we report on the largest series of IgM-secreting-DLBCL, from a multicentre Italian study. METHODS: The observational and biological study was approved by the Ethical Committee of the AUO Sant'Andrea, Italy. Enrolment criteria were: DLBCL with an associated IgM paraprotein diagnosed between 1st January 2010 and 31st December 2018 (IgM-secreting). Data were collected both prospectively and retrospectively from 17 Centres participating in the study. In addition, histopathology samples were centrally revised for immunohistochemistry (IHC) and FISH analyses. The control group (CTRL) consisted in a series of consecutive DLBCL, without an associated IgM-paraprotein (diagnosed between 01/01/2013 and 30/06/2016, enrolled in the Lymphoma Registry of the Lazio region (ReLLi Network). Last follow-up was carried out on 31st December 2019. RESULTS: 569 DLBCL cases were enrolled: 102 (17.9%) were IgM-secreting; 48 (8.4%) had a non-IgM paraprotein (IgA, IgG, or other), and 414 (72.7%) had no associated paraprotein (CTRL). IgM-secreting cases within the consecutive DLBCL patients enrolled in the ReLLi Registry were 41/466 (8.8%, 95CI 6.4-11.7%) while non IgM-paraprotein DLBCL cases were 11/466 (2.4%, 95CI 1.2-4.2%). The median level of IgM paraprotein was 17gr/L (range: &amp;lt;1-84gr/L); 83/102 (81.3%) were IgMk and 23/102 (22.5%) IgML respectively. The IgM-secreting group differed from the CTRL because the following characteristics were significantly more frequent: 1] age&amp;gt;60 (p=.001); 2] advanced stage (p&amp;lt;.001); 3] PS≥2 (p=.001); 4] LDH&amp;gt;UNL (p=.008) ; 5] ≥2 Extra-nodal sites involved (p&amp;lt;.001) ; 6] IPI 3-5 (p&amp;lt;.001); 7] central nervous system (CNS) involvement at diagnosis or relapse (p&amp;lt;.001); 8] lower rate of complete remission(CR) at the end of induction immunochemotherapy (p&amp;lt;.001). Conversely, no differences were observed for: sex, B-symptoms, HCV and HBV status, bulky disease, age≥80 years, and for transformation from low-grade lymphoma. PATHOLOGICAL AND MOLECULAR FEATURES: Paraffin tissue from 74 CTRL and 69 IgM-secreting was suitable for immunohistochemistry (IHC). The non-GCB subtype, based on Hans algorithm, was prevalent in the IgM-secreting (p=.005). No difference in BCL2 expression alone or in MYC and BCL2 double expression was observed within groups. In 48/63(76%; 95CI: 64-86%) IgM-secreting cases, both the IgM heavy and the corresponding kappa or lambda light chain protein expression were detected in the cytoplasm of the neoplastic clone. FISH analyses for MYC, BCL2 and BCL6 genes rearrangements performed in 25 IgM-secreting cases with either expression of MYC protein or a GC-phenotype showed no evidence of double or triple hits (DH/TH). TREATMENT: in the IgM-secreting group more patients were treated with RCOMP and with less intensive approach than the CTRL (p&amp;lt;.001). SURVIVAL: The median follow-up time was 46 months (95CI= 44-49; range 18-101) with 130 events and an incidence rate x100 person/year of 7.22 (95%CI 6.08-8.58) and a 5-yr OS of 76% (95CI 72-79%). The 5-PFS was 61% (95CI 57-65%). In univariable analysis age&amp;gt;60, B-symptoms, bulky disease, IPI &amp;gt;low risk and IgM-secreting IgM showed a worse survival (all with p&amp;lt;0.001). Also, the IgM-secreting group, showed a worse survival compared to the DLBCL with an associated IgG/IgA paraprotein (p&amp;lt;0.001). Adjusting in multiple Cox regression, IgM-secreting with IPI, gender, bulky and B-symptoms, maintain a higher risk of death either in the all cohort (HR 1.93, 95CI 1.34-2.78, p&amp;lt;0.001) or in patients with age&amp;lt;80 (HR 1.71, 95CI 1.16-2.54, p=0.007). Noteworthy, a survival sub-analysis showed that the 12/69 (17.4%) IgM-secreting with a GC-type had a better OS (9=0.008) and PFS (p=0.002) compared to the 57/69 (82.6%) IgM-secreting with a non-GC-type. CONCLUSION: Our data confirm that IgM-secreting DLBCL: 1) represents a sizable proportion of non-DH DLBCL; 2) have poor prognostic features and 3) have mostly a non-GC phenotype. Furthermore, IgM secretion appears to be an independent prognostic factor for both PFS and OS. Studies to define the biological features of this new subset are ongoing. Disclosures Cantonetti: Mundipharma: Consultancy; Takeda: Consultancy; Vifor: Consultancy; Roche: Consultancy. Re:BerGenBio ASA: Research Funding. Abruzzese:Incyte: Consultancy, Membership on an entity's Board of Directors or advisory committees; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees; Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees; Bms: Honoraria.

Abstract C098: Racial disparity in the prevalence of BCL-2 expression and associations with breast cancer survival in the Breast Cancer Care in Chicago study

Abstract Background: BCL-2 is an antiapoptotic protein that regulates apoptosis and has been associated with poor prognosis for many cancer sites. Despite a function that encourages tumorigenesis, increased BCL-2 expression has been associated with improved clinical outcomes in ER+ breast cancers, with mixed results observed for ER/PR- cancers. BCL-2 has been hypothesized to be an indicator of intact and fully functioning hormone receptor pathways and has also been hypothesized to facilitate cell death after treatment through mitochondrial priming. BCL-2 expression correlates strongly with hormone receptor expression; therefore, its association with better prognosis may be due to the strong effect ER/PR status has on survival, or due to its own biologic mechanism. To date, no studies have been conducted on racial differences in BCL-2 expression. We characterized racial/ethnic differences in BCL-2 expression and the association of BCL-2 expression with breast cancer specific survival among participants in the Breast Cancer Care in Chicago Study (BCCC). Methods: The BCCC was a cross-sectional study of 989 recently diagnosed non-Latina (nL) White, nL Black and Latina breast cancer patients diagnosed with a first primary breast cancer aged 30-79 in Chicago from 2005-2008. Tumor tissue was available for BCL-2 immunohistochemical staining for 264 patients. BCL-2 staining scores of 0 were classified as negative and all positive scores (1D, 1H, 2D, 2H, and 3) were classified as positive. Cox proportional hazards models were conducted to estimate the association of BCL-2 expression with breast cancer-specific death. Results: Roughly three-fourths of tumors (77%) stained positive for BCL. The prevalence of positive BCL staining varied by race/ethnicity (p=0.007), being highest for nL Whites (87%) and lowest for nL Blacks (68%); variability in BCL staining appeared to be limited to ER/PR-negative tumors (N=53), for which prevalence of positive BCL staining appeared to be much higher for nL whites than for minority patients (44% vs. 16%, p=0.054). BCL-2 expression was associated with a strong protective effect on BC survival in age and race-adjusted models (HR=0.40, 95% CI: 0.19, 0.84) and BCL-2 expression remained strongly positively associated with protection from breast cancer death with additional adjustment for ER/PR status (HR=0.33, 95% CI: 0.12, 0.89). Within subgroups defined by ER/PR status, BCL2 expression was qualitatively associated with better survival for both ER/PR positive and ER/PR negative subtypes, although sample size was insufficient to conduct a formal stratified analysis. Conclusions: BCL-2 appears to be an independent predictor of improved prognosis for breast cancer even after controlling for ER/PR status. The apparently higher prevalence of BCL2 expression for nL White vs. nL Black patients may provide etiologic clues regarding disparities in BC survival. Citation Format: Jibril M. Alim, Kent Hoskins, Abeer M. Mahmoud, Virgilia Macias, Andre Kadjascy-Balla, Garth H. Rauscher. Racial disparity in the prevalence of BCL-2 expression and associations with breast cancer survival in the Breast Cancer Care in Chicago study [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr C098.

Efek Proteksi dari Terapi Oksigen Hiperbarik terhadap Ekspresi Bcl-2 Miometrium Rattus norvegicus Bunting yang Terinfeksi oleh Tachyzoite Toxoplasma gondii

Hyperbaric Oxygen Therapy (HBOT) can increase oxygen delivery to tissues and stimulate the formation of H2O2 as a secondary messenger for phosphorylation of nuclear factor kappa beta (NF-kB) which plays an important role in the transcription of the anti apoptotic gene. This study aimed to determine the effects of Hyperbaric Oxygen Therapy (HBOT) in enhancing the expressions of Bcl-2 in the myometrium of pregnant rats infected by Toxoplasma gondii. This study was an experimental study with a randomized control group of post-test only and designed by 37 pregnant Rattus norvegicus Sprague Dawley. Randomly, the rats were divided into four groups. Group A is infected pregnant rats that exposed by 10 sessions of HBOT 2.4 ATA in 3x30 minutes. Group B is non-infected pregnant rats and exposed by 10 sessions of HBOT 2.4 ATA in 3x30 minutes. Group C is infected pregnant rats without any exposure. Group D is non-infected pregnant rats without any exposure. Each infected pregnant rat was given a 103 tachyzoite of T.gondii by intraperitoneal injection. Bcl-2 expressions were measured through immunohistochemistry. All data were analyzed using ANOVA test through SPSS 21 program application. There was a significant difference in Bcl-2 expression between Group A and Group C because p&lt;α (p&lt;0.017). HBOT can increase the expression of Bcl-2 from infected and not infected rat myometrium, in the provision of HBOT 2.4 ATA for 3x30 minutes, twice a day for 5 days.

Prognostic Significance of BCL2 Protein in Diffuse Large Cell Lymphoma of Head and Neck; Relation to Response to Chemotherapy

Diffuse large B cell lymphoma (DLBCL) is a heterogeneous disease that displays a highly variable clinical outcome. It is a neoplasm of large transformed B cells with a diffuse growth pattern. DLBCL is the most common type of non-Hodgkin’s lymphoma (NHL) (31% of all cases). Approximately half of patients with DLBCL are cured with current chemotherapy regimens. The purpose of this study was to evaluate BCl2 expression in 45 patients diagnosed with DLBCL of head and neck region and correlate the level of its immunohistochemical expression with different clinicopathological variables with emphasis upon patients’ age, gender, nodal or extra-nodal location of lymphoma, patients’ response to chemotherapy, progression-free survival (PFS) and overall survival (OS). A retrospective analysis of 45 patients diagnosed to have DLBCL. A cut off value of ≥ 50% protein expression denoted BCL2 positivity. Out of 45 cases, 36 cases (80%) revealed BCL2 positive expression and 9 cases (20%) were BCL2 negative. We found statistically significant differences in BCL2 expression regarding different patients’ responses to chemotherapy, patients’ OS and PFS (p ≤ 0.05). No statistically significant differences in BCL2 expression regarding the patients’ Ann Arbor clinical stage, age group and tumor site (nodal or extra-nodal, p > 0.05) using the Chi-square test. BCL2 expression was analyzed in relation to 5 years OS and PFS using Kaplan Meier curves and Log Rank test for survival analysis. Cases that demonstrated BCL2 positivity revealed shortened OS and PFS with highly statistically significant differences among the studied variables (p = 0.000). We also found that patients who respond well to the chemotherapeutic regimen had negative BCL2 expression, the differences were statistically significant (p = 0.015). In conclusion, BCL2 expression could be considered a predictor for patients’ chemotherapeutic response, OS and PFS.

Baicalin protects mouse testis from injury induced by heat stress.

Heat stress has been documented to reduce reproductive performance of female animals through injury to germ cells, with few studies available in male animals. The objectives of this study were to evaluate protective effects of baicalin on testicular tissue damage of mice subjected to heat stress and its related mechanisms. In this experiment, A total of forty mice were divided into four groups, including control group (C), baicalin group (B), heat stressed group (H) and heat stress with baicalin treatment (H+B) group. Morphological changes, activities of antioxidant enzymes and apoptosis-related parameters in the mice testes tissue were monitored. The results showed that the process of spermatogenesis in mice testis was impaired and the cellular apoptosis increased due to acute heat stress at 41°C. Interestingly, the tissue damage was alleviated with the significant (P < 0.05) increase in the activities of SOD, CAT and GSH-Px enzymes, decrease (P<0.05) in MDA content and number of cellular apoptosis recorded in mice of H+B group compared with those in mice from H group. In addition, the Fas, FasL and P-JNK protein expressions were significantly (P<0.05) increased; and apaf-1, caspase-3, -9 were slightly expressed in the H group, while there was no difference in Bcl-2 expression, compared with C, B and H+B groups. The above results clearly indicate that heat stress induces macroscopic/apoptotic and oxidative changes in the testicular tissue of mice; these changes are alleviated by Baicalin through increasing anti-oxidative enzyme activities and possibly through blocking Fas/FasL pathway.

Venetoclax in Combination with FLAG-IDA Chemotherapy (FLAG-V-I) for Fit, Relapsed/Refractory AML Patients: Interim Results of a Phase 1b/2 Dose Escalation and Expansion Study

Venetoclax in Combination with FLAG-IDA Chemotherapy (FLAG-V-I) for Fit, Relapsed/Refractory AML Patients: Interim Results of a Phase 1b/2 Dose Escalation and Expansion Study

The Effect of Se'i (Smoked Beef) Toward the Improvement of the Bcl-2 Protein expression on Colon Cells of Balb/c strain Mice as a Carcinogenesis Indicator

Se'i is smoked beef with added nitrite which can precipitate the Bcl-2 protein expression. The purpose of this study was to analyze the effect of se'i toward the improvement of the Bcl-2 protein expression on colon cells of Balb/c strain mice as an indicator of carcinogenesis. This research is true experimental with completely randomized design. Samples were male Balb/c mice, weighing 23.8 gram which was 36 mices, divided into 4 treatment groups P1: Mice were given se'i without nitrite with weight 8.840 mg for 28 days, P2: Mice were given se'i containing nitrite from the modern industry weighing 8.840 mg for 28 days, P3: Mice were given nitrite se'i from home industry weighing 8.840 mg for 28 days and K: Mice fed only standardize feed, and drinking water for 28 days. The examination was done by using immunohistochemical techniques to see the expression of a Bcl-2 protein. The result showed that there was a significant difference of Bcl-2 protein expression on colon cells of male Balb/c strain mice which are given se'i. The LSD test showed that there was a difference of Bcl-2 protein expression between P3, P1 and K. The new finding of this study is the provision of se'i causing the differences in Bcl-2 expression in all treatment groups.

IMMUNOHISTOPATHOLOGICAL ROLE OF BCL2 AND P53 GENE EXPRESSION IN HELICOBACTER PYLORI CYTOTOXIN-ASSOCIATED GENE A POSITIVE VERSUS CYTOTOXIN-ASSOCIATED GENE A NEGATIVE ANTRAL PREDOMINANT NON-ATROPHIC GASTRITIS IN IRAQI PATIENTS

ABSTRACTObjectives: To determine the role of Bcl2 and p53 gene expression in the pathogenesis of antral-predominant non-atrophic gastritis (APNG) accordingto Helicobacter pylori (HP) cytotoxin-associated gene A (CagA) status.Methods: Multiple antral biopsies were taken from 78 patients for rapid urease test, histopathology, Bcl2 and P53 immunohistochemistry and HPCagA in situ hybridization.Results: CagA was detected in 74.35% cases. There was no significant difference in Bcl2 expression among CagA+ and CagA− APNG cases. Significantdifferences in polymorphonuclear neutrophils (PMNs) and lymphocytes grades were detected among CagA+ and CagA− APNG cases. There was nosignificant correlation among patients’ age, inflammatory infiltrates, Bcl2, p53 expression. CagA has positive correlation with p53 (p=0.001), PMNsgrade (p=0.027), lymphocytes grade (p=0.003), inflammation intensity (p=0.006), and inflammation activity (p=0.007). Bcl2 has no significantcorrelation with p53, CagA, PMNs, and lymphocytes indexes. P53 expression has significant correlation with PMNs and lymphocyte grades (p=0.000),inflammation intensity (p=0.003), and inflammation activity (p=0.002). PMNs grade has positive correlation with lymphocyte grade, inflammationintensity, and activity (p=0.000). Lymphocyte grade has a significant correlation with inflammation intensity and activity (p=0.000). Inflammationintensity has a significant correlation with inflammation activity (p=0.000).Conclusions: CagA cytotoxin has direct effect on P53 gene and indirect effect on Bcl2 gene expression in APNG cases. Bcl2 and P53 expression donot get affected by patient’s age and gender. PMNs grade, lymphocytes grade, inflammation intensity, and inflammation activity affected directly byP53 and CagA cytotoxin expression and indirectly by Bcl2 expression. The balance of P53-Bcl2 pathways play a vital role in pathogenesis of HP andCagA-induced APNG.Keywords: Antral-predominant non-atrophic gastritis, Bcl2, p53, Helicobacter pylori, Cytotoxin-associated gene A.

Investigation of the effect of safranal and crocin pre-treatment on hepatic injury induced by infrarenal aortic occlusion.

Ischemia-reperfusion (IR) injury of the liver is an unresolved problem that occurs during certain surgical approaches, including hepatic, cardiac and aortic operations. In this study we aimed to investigate whether crocin and safranal had protective effects on liver IR injury induced in an infrarenal aortic clamping (IRAC) model. Male Wistar-Albino rats (n=32) were divided into four groups with 8 animals each as follows: Sham, IR, IR+crocin, and IR+safranal. The infrarenal aorta (IRA) was clamped for 60min for the ischemic period and allowed to reperfuse for 120min. Blood and tissue samples were collected for biochemical, histological and immunohistological analysis. Plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were found to be significantly higher in the IR group than the sham group (respectively; p=0.015, p<0.001). There were significant differences between the IR group and the IR+crocin group or the IR+safranal group in AST levels (respectively; p=0.02, p<0.001). ALT showed a significant decrease in the IR+crocin group compared to the IR group (p<0.05). We also observed histopathological changes among the groups. Bax and Caspase-3 expression in the IR group was remarkably higher than in the other groups. Caspase-3 and Bax expression in the IR+crocin and the IR+safranal groups were significantly lower than in the IR group. Nevertheless, there were no significant differences in BCL2 expression among the groups. IRAC is a cause of IR injury in the liver. This study showed that crocin and safranal have protective effects on IR induced liver injury.

Interval of IL-2 Administration Has a Major Impact on Treatment Efficacy for Regulatory T Cell Expansion and Homeostasis

IL-2 has a critical role in the immune homeostasis by expansion and maintenance of regulatory T cell (Treg). We previously demonstrated that low-dose IL-2 administration preferentially increased Treg in patients with active chronic GVHD and resulted in clinical improvement of the symptoms. In these years, the tolerogenic effects of IL-2 have been tested in the setting of various autoimmune-based diseases by many clinical trials. However, the schedules of IL-2 administration in each trial are different and the optimal strategy for expansion and maintenance of Treg is still unclear. To tackle this issue, we examined the impact of IL-2 dose intervals on Treg and the optimization for the induction and maintenance therapy by using murine IL-2 therapy model. CD4+CD25+Foxp3+ Treg were analyzed comparing with CD4+CD25-Foxp3- conventional CD4 T cells (Tcon) and CD8+ T cells. The expressions of Ki-67 and Bcl-2 in each subset were also examined. First, we performed the experimental allogeneic BMT, in which model 1x10E6 spleen cells (CD45.2) and 5x10E6 T cell depleted bone marrow cells (CD45.1) from C57BL/6 donors were transplanted into lethally-irradiated B6D2F1 recipients (CD45.2). To explore the optimum dose-interval for the increase of Treg from the baseline level as the induction therapy, we administered 5000 IU of IL-2 to recipients subcutaneously once (Induction-A), twice (Induction-B), four (Induction -C) or seven (Induction-D) times a week from day 35 to 49 after transplant. On day 50 after transplant, peripheral blood and spleen cells of mice from each group were harvested and CD45.1+H-2Kd- Donor bone-marrow derived lymphocytes were evaluated. The absolute number and %Treg of CD4 T cells, and the Ki-67 and Bcl-2 expression in Treg were compared with each frequency of administration. To increase Treg from baseline, the daily administration (Induction-D) provided the best Treg response among the tested groups and there was significant difference between control group and group I-D (%Treg: 10.5 % vs. 16.7 %, p<0.05). Treg proliferation was positively related to the frequency of IL-2 administration (%Ki-67+ cells: 17.7%, 17.2 %, vs. 9.9%, 9.0% and 7.8%, respectively, p<0.05). Secondly, we investigated the optimum dose-interval for the maintenance of expanded Treg level after the initial inductive IL-2 administration. We expanded Treg by daily administration of IL-2 to B6 mice for 2 weeks, thereafter we administrated IL-2 twice (Maintenance-A), four (Maintenance-B), or seven (Maintenance-C) times a week from day 14 to 21 for the maintenance. Of interest, to maintain expanded Treg level after the induction IL-2 therapy, Treg in cohorts of Maintenance-B and Maintenance-C were significantly higher than in cohort Maintenance-A (mean 9.7%, 10.2% vs 7.1%, respectively) and there was no significant differences between cohorts Maintenance-B and Maintenance-C, suggesting daily administration is not necessary and intermittent administration within the threshold may work for the maintenance of expanded Treg level after the intensive IL-2 administration. Treg proliferation of cohort Maintenance-C was significantly higher than that of the other cohorts (mean %Ki-67: 19.8 %, 20.5 % vs. 23.9 %, p<0.05). There were no significant difference in Bcl-2 expression in Treg among these cohorts but seemed negatively related to the frequency of IL-2 administration. These data suggested that daily administration of IL-2 seems to be optimal for expansion of Treg for induction therapy. In contrast, to maintain the expanded Treg, daily administration may not be required but less frequent times of administration within the threshold could be preferable. Taken together, the interval of IL-2 administration should be a major factor for Treg homeostasis as well as IL-2 dosage. Our data provide important information for developing therapeutic strategies to modulate Treg homeostasis in vivo and promote immune tolerance after transplant. DisclosuresNo relevant conflicts of interest to declare.

Expression of Fas (CD95) and Bcl-2 in peripheral blood mononuclear cells in patients with chronic HCV and schistosomiasis

A high incidence of viral persistence and progression to chronic hepatitis are characteristic features of HCV infection. The aim of this case control study was to investigate the expression of the apoptosis related proteins Fas (CD95) and Bcl-2 in peripheral blood mononuclear cells (PBMCs) from patients with chronic HCV and schistosomiasis, and to evaluate their contribution in the development and maintenance of the different clinical forms of these diseases. The level of Fas and Bcl-2 expression in PBMCs was detected using flow cytometry in 85 cases in this study, including chronic HCV (40 patients) and schistosomiasis (25 patients) in addition to 20 healthy controls. Results showed that the increased apoptotic level correlated with increased Fas expression on the surface of PBMCs in chronic HCV patients (p = 0.001) which did not show increased expression of the anti-apoptotic factor Bcl-2 (p = 0.19). Apoptotic PBMCs in intestinal schistosomiasis patients showed increased expression of Fas (p = 0.001) but again did not exhibit a difference in Bcl-2 expression (p = 0.61) when compared to either hepatoplenic schistosomiasis or control patients. The Fas expression was highly significant in HCV > schistosomiasis > control (p < 0.002) while non-significant difference was found between the 3 groups as regards to Bcl-2 expression (p > 0.12). It was concluded that the detection of apoptosis of PBMCs in patients with chronic HCV was associated with increased expression of apoptotic markers in comparison with schistosomiasis patients, which may be an indicator of disease progression and severity. Modulation of this process may offer valuable methods of HCV therapy.

Cyclooxygenase-2 and B-cell lymphoma-2 expression in cystitis glandularis and primary vesicle adenocarcinoma

BackgroundAlthough cystitis glandularis (CG) is a common benign urinary bladder epithelial abnormality, it remains unclear whether CG is a premalignant lesion. Cyclooxygenase-2 (COX-2) and B-cell lymphoma-2 (Bcl-2) overexpression has recently been reported as a potential tumor initiator or promoter. We evaluated and compared COX-2 and Bcl-2 expression in CG, chronic cystitis (CC), and primary vesicle adenocarcinoma (ADC) tissues.MethodsWe conducted a retrospective study to investigate COX-2 and Bcl-2 levels in CG and ADC. We obtained tissue samples from 75 patients (including 11 cases of CC, 30 typical cases of CG (CGTP), 30 cases of intestinal CG (CGIT), and 4 cases of ADC) between 1989 and 2009 from the Surgical Pathology Archives of the No. 2 People’s Hospital of Zhenjiang, affiliated with Jiangsu University. COX-2 and Bcl-2 immunohistochemical staining was performed on all tissues. Nine normal bladder epithelial specimens were evaluated as control samples. Correlations between COX-2 and Bcl-2 expression in CG were also analyzed.ResultsCOX-2 and Bcl-2 expression was higher in the ADC group compared to other groups (p < 0.05). COX-2 and Bcl-2 levels were higher in the CGIT group compared to the CGTP group (p = 0.000 for both). The CGIT and CGTP groups both showed higher COX-2 expression compared to the CC group (p = 0.000 for both). There was no difference in Bcl-2 expression between the CGTP and CC groups (p = 0.452). Additionally, the difference in COX-2 and Bcl-2 expression between the control and CC groups was also insignificant (p = 0.668 and p = 0.097, respectively). Finally, we found that COX-2 and Bcl-2 levels were positively related (r = 0.648, p = 0.000).ConclusionCOX-2 and Bcl-2 overexpression in the CG group suggests that CG, particularly the intestinal type, may be a premalignant lesion that converts into a tumor in the presence of carcinogens.

P53 and bcl2 expression in malignant and premalignant lesions of uterine cervix and their correlation with human papilloma virus 16 and 18

Background and Objective:Persistent high risk human papilloma virus (HPV) infection is probably the best predictor of increased risk of cervical cancer, but expression of certain markers of cell proliferation and apoptosis have been studied. The present study was conducted to evaluate the expression of p53 and bcl2 in premalignant and malignant lesions of cervix and its correlation with HPV type 16 and 18.Materials and Methods:The study comprised of 35 cases (including 24 prospective cases and 11 retrospective cases) of premalignant and malignant lesions of the cervix. Slides were stained with Hematoxylin and Eosin and p53, bcl2 (immunohistochemistry), HPV 16 and HPV 18 (in situ hybridization).Results:p53 positivity was seen in 8/19 (42.1%) cases of cervical intraepithelial neoplasia (CIN) and 8/16 (50%) cases of carcinoma cervix, the difference not significant statistically. The difference in bcl2 expression in CIN versus carcinoma cervix (84.21% vs. 43.75%) was statistically significant (P = 0.030). There was no significant difference between p53 and bcl2 expression and the stage and grade of the tumors. Seven out of 19 cases of CIN (36.84%) were positive for HPV 16/18 infection and 8/16 cases (50%) of carcinoma cervix were HPV positive (P = 0.628).Conclusions:No significant association was found between HPV 16/18 infection and p53 and bcl2 expression in premalignant and malignant lesions of uterine cervix. Although, bcl2 staining showed a significant difference between CIN and carcinoma cervix, a larger case series is required to assess the association between HPV infection and overexpression of p53 and bcl2 proteins in these lesions.