Monoclonal Ki-67 Research Articles

Mechanism of Paeoniae Radix Rubra and Aconiti Lateralis Radix Praeparata herbal pair in promoting hepatocellular regeneration in chronic acute liver failure based on HGF/PI3K/Akt signaling axis

Based on the hepatocyte growth factor(HGF)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt) signaling axis, this study investigated the therapeutic effect of Paeoniae Radix Rubra and Aconiti Lateralis Radix Praeparata(PRR-ALRP) her-bal pair on acute-on-chronic liver failure(ACLF) rats and its impact on hepatocellular regeneration. The rat model of ACLF was constructed by subcutaneous and tail vein injection of bovine serum albumin combined with intraperitoneal injection of lipopolysaccharides(LPS)+D-galactosamine(D-GalN). The rats were divided into normal control(NC) group, model(vehicle) group, PRR-ALRP(5.85 g·kg~(-1)) group, and hepatocyte growth factor granules(HGFG, 4.05 g·kg~(-1)) group. Hematoxylin-eosin(HE) staining was used to observe pathological changes in rat liver tissues. Serum alanine aminotransferase(ALT), aspartate transaminase(AST), and total bilirubin(TBIL) were detected using an automatic biochemical analyzer. The levels of interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) inflammatory factors were detected by ELISA. Immunofluorescence staining was used to detect the positive expression of proliferating cell nuclear antigen(PCNA), antigen identified by monoclonal antibody(Ki67), and cell cycle protein B1(CyclinB1). Real-time fluorescence-based quantitative polymerase chain reaction(RT-qPCR) and Western blot were used to detect the mRNA and protein expression levels of HGF, growth factor receptor-bound protein 1(Gab1), PI3K, Akt, phosphorylated PI3K(p-PI3K), and phosphorylated Akt(p-Akt). The results showed that compared with the vehicle group, the PRR-ALRP group had reduced liver tissue pathological scores, improved liver function, and reduced inflammatory response, with enhanced PCNA, Ki67, and CyclinB1 fluorescence expression. Furthermore, compared with the model group, the PRR-ALRP group showed upregulated expression of HGF and Gab1 proteins, as well as activation of PI3K and Akt phosphorylation. These findings suggest that PRR-ALRP herbal pair exerts anti-liver failure effects by alleviating hepatocyte inflammatory damage and promoting hepatocellular regeneration, and its specific regulatory mechanism may be related to the activation of the HGF/PI3K/Akt signaling pathway.

Relationship between PIWIL1 gene polymorphisms and epithelial ovarian cancer susceptibility among southern Chinese woman: a three-center case–control study

ObjectiveTo investigate the potential correlation between piwi-like RNA-mediated gene silencing 1 (PIWIL1) polymorphisms and susceptibility to epithelial ovarian cancer (EOC).MethodsA case–control study was conducted to evaluate the susceptibility of EOC using multinomial logistic regression analysis. The study analyzed the relationship between five functional single nucleotide polymorphisms (SNPs) in the PIWIL1 gene and EOC risk. Genotyping of 288 cases and 361 healthy samples from South China was identified using a TaqMan assay. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to estimate the relationship between the five selected SNPs and EOC susceptibility.ResultsAmong the five SNPs analyzed, the rs10848087 G > A and rs7957349 G > C variants significantly increased the susceptibility of EOC, rs10773771 C > T was associated with a decreased risk of EOC, while the rs35997018 and rs1106042 variants were not in Hardy–Weinberg equilibrium (p < 0.05). The rs10848087 G > A was significantly associated with increased risk of EOC in individuals with metastasis, FIGO stage I and III, low and high pathological grade, tumor numbers ≤ 3 and > 3, tumor size > 3 cm and ≤ 3 cm, pregnant more than 3 times, pre-menopausal status, and strong positive expression of ER (estrogen receptor), PR (progesterone receptor), PAX8 (paired-box 8), wild-type p53 (tumor protein 53), WT1 (Wilm’s tumor gene), P16 (cyclin-dependent kinase inhibitor 2A). In addition, rs10848087 G > A enhanced the EOC risk of cases with negative/mild positive expression of wild p53 and Ki67, and with or without mutant p53 expression. The rs7957349 G > C variant was linked to an increased risk of EOC in subgroups with certain characteristics, including age equal or less than 53 years, metastasis, clinical stage I, low pathological grade, tumor number, tumor size, pregnant times, post-menopause, pre-menopause, and strong positive expression of wild p53 and Ki67 (Antigen identified by monoclonal antibody Ki-67), as well as without mutant p53 expression. The rs10773771 CT/TT alleles were identified to have a protective effect on EOC in women aged 53 years or older, as well as in cases with metastasis, advanced clinical stage, high pathological grade, multiple tumors, tumor size equal to or less than 3 cm, history of pregnancy, post-menopausal status, and strong positive expression of ER, PR, wild-type p53, PAX8, WT1, P16, and Ki67. Furthermore, rs10773771 CT/TT also showed a protective effect in patients with negative or mildly positive expression of PR, PAX8, wild-type p53, WT1, and P16, as well as positive expression of mutant p53. Compared to the reference haplotype GCG, individuals harboring haplotypes GTG were found to have a significantly decreased susceptibility to EOC. PIWIL1 was significantly expressed in the thyroid, pituitary, and adrenal glands with rs7957349 CC alleles.ConclusionsPIWIL1 rs10848087 and rs7957349 were associated with increased risk of EOC, while rs10773771 may have a protective effect against EOC. These genetic variants may serve as potential biomarkers for EOC susceptibility in the South China population.

Study of Ki67 in odontogenic benign tumors

The aim of the present study was to investigate the Ki67 expression level and to measure the cell proliferation index (Ki67) in odontogenic benign tumors.This was an analytical cross-sectional study of odontogenic benign tumors. The sampling was non-probabilistic with an exhaustive recruitment including all biopsy or surgical excision specimen of odontogenic benign tumors. Ki67 immunohistochemistry was performed on histological sections of paraffin-fixed tissues of 3µ thickness with the Ki67 (MM1) monoclonal antibody. The studied variables were sociodemographic, clinical and histopathological. The data have been analysed with SPSS 20.0 software.: Ameloblastoma represented 50.9% of cases and cemento-osseous dysplasia 36.4%. Among the 28 ameloblastoma cases, the 15 (53.6%) were plexiform type, the 7 (25%) follicular type and the 6 (21.4%) unicystic type. The percentage of Ki67 labelled cells was less than 1% in 74.6% (n=41) of the tumors and was equal to 5% in 12.7% (n=7) of cases. A Ki67 law labelling was found in 87.3% (n=48) of tumours and a negative labelling in 12.7% (n=7) of cases. The odontogenic benign tumors had a weak Ki67 expression level and a low epithelial cell proliferation index. The Ki67 could not therefore be used as a predictive biomarker of tumor cell proliferation and the risk of recurrence in odontogenic benign tumors.

Application effect of preoperative chemoradiotherapy combined with rehabilitation nursing in patients with rectal cancer surgery

ABSTRACT To study the effect of preoperative chemoradiotherapy combined with rehabilitation nursing in patients with rectal cancer surgery. 106 cases of rectal cancer patients in our hospital were selected. 53 cases in each group were treated with surgical treatment combined with rehabilitation nursing treatment and preoperative radiotherapy and chemotherapy combined with surgical treatment and rehabilitation nursing treatment in the study group. The T stage (ypT) and N stage (ypN) downgrading rates of serum carcinoembryonic antigen (CEA) and carbohydrate antigen (CA)19–9 were compared between the two groups after treatment. The 5-year cumulative survival rate, recurrence rate and the positive rate of Bax and antigen identified by monoclonal antibody Ki-67 (Ki-67) expression were detected. T stage downgrading rate and N stage downgrading rate were 77.36% (41/53) 35.85% (19/53) in control group and 94.34% (50/53) 64.15% (34/53) in research group, respectively. The CEA and CA19–9 levels measured at the end of surgery and one month after nursing in both groups were lower than those before treatment. After treatment, scores of quality of life indicators in both groups increased. The positive rates of Bax and Ki-67Ki-67 were significantly different between the two groups after treatment (P < 0.05). Preoperative chemoradiotherapy combined with rehabilitation nursing has obvious effect on patients with rectal cancer surgery, and has obvious advantages in inhibiting tumor growth, destroying tumor survival immune environment and reducing surgical complications, which can improve the prognosis and is worthy of clinical application. It could provide a potential treatment for patients with rectal cancer.

Effect and mechanism of longkui yinxiao soup in treating psoriasis in mice.

Objective: Longkui Yinxiao Soup is a traditional Chinese medicine formula used to treat psoriasis for decades. Although Longkui Yinxiao Soup showed promising efficacy in clinical practice, the regulatory mechanisms of Longkui Yinxiao Soup remain elusive. This study aimed to explore the underlying mechanisms of Longkui Yinxiao Soup in a psoriasis-like mouse model. Methods: Longkui Yinxiao Soup was quality controlled by determining the contents of imperatorin and rhoifolin using high-performance liquid chromatography. The imiquimod-induced psoriasis-like mouse model was used to study the therapeutic effect and mechanism of Longkui Yinxiao Soup. The histopathological skin changes were observed by hematoxylin and eosin staining; the infiltration of proliferating proteins, proliferating cell nuclear antigen and Ki67, in skin tissues were observed by immunohistochemical analysis; and the inflammatory factors such as interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-23, and IL-17 in serum were detected using enzyme-linked immunosorbent assay. RNA sequencing and bioinformatic analysis were used to predict the mechanism of LYS against psoriasis. mRNA expressions of p38, extracellular regulated protein kinases (ERK), mitogen-activated protein kinase 3 (MEK3), mitogen-activated protein kinase 6 (MEK6), RAP1 GTPase activating protein (Rap1gap), and Rap1 were determined using real-time quantitative polymerase chain reaction. The expression levels of proteins related to Rap1-mitogen-activated protein kinase signaling pathways were measured by Western blotting. Results: A quality-control method for Longkui Yinxiao Soup was successfully established using imperatorin and rhoifolin as content determination indexes. Longkui Yinxiao Soup significantly ameliorated the psoriatic symptoms in mice. The serum levels of inflammatory cytokines such as IL-6, TNF-α, IL-23, and IL-17 were decreased, and the expression levels of antigen identified by monoclonal antibody Ki67 (Ki67) and PCNA in skin tissues were downregulated. Moreover, the inhibition of Rap1-MAPK signaling pathways by Longkui Yinxiao Soup was detected. Conclusion: This study confirmed the antipsoriatic activity of Longkui Yinxiao Soup in psoriasis-like mice. This might be due to the inhibition of inflammatory factor secretion, keratinocyte proliferation, and the Rap1-MAPK signal pathway.

Effect of ethanol extract from Chrysanthemum cinerariifolium leaves on Ki-67 proliferation and dysplasia severity in a rat model of oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is a malignant tumor that can rapidly infiltrate the oral epithelial tissue and cause high mortality worldwide because the available therapies are less effective. Chrysanthemum cinerariifolium leaf contains secondary metabolites as anti-inflammatory, antioxidant, anticancer, and antimutagenic. The study aimed to analyze the ethanolic extract of C. cinerariifolium leaf in reducing proliferation (Ki-67) and the degree of dysplasia in OSCC rats. This study used male Sprague Dawley induced by 7,12-dimethylbenz(a)anthracene (DMBA) 0.5% and divided into five treatment groups, namely positive control/C+ (sick), negative control/C- (healthy), and treatment group induced with DMBA and given extract C. cinerariifolium leaf with successive doses of T1, T2, and T3 (50, 100, and 200 mg/kg bw). The oral epithelium was stained with hematoxylin and eosin and immunohistochemically stained with a Ki-67 monoclonal antibody. The statistical analysis utilizes the one-way analysis of variance test. The results showed that T1 at a dose of 200 mg/kg bw could significantly reduce Ki-67 expression and the degree of oral epithelial dysplasia (OED; p < 0.05) close to healthy controls. The conclusion shows that C. cinerariifolium leaf extract can be a therapy against OSCC by decreasing cell proliferation and the degree of OED.

Focal adhesion alterations in G0-positive melanoma cells.

Melanoma is a highly heterogeneous malignant tumor that exhibits various forms of drug resistance. Recently, reversal transition of cancer cells to the G0 phase of the cell cycle under the influence of therapeutic drugs has been identified as an event associated with tumor dissemination. In the present study, we investigated the ability of chemotherapeutic agent dacarbazine to induce a transition of melanoma cells to the G0 phase as a mechanism of chemoresistance. We used the flow cytometry to analyze cell distribution within cell cycle phases after dacarbazine treatment as well as to identifyG0 -positive cells population. Transcriptome profiling was provided to determine genes associated with dacarbazine resistance. We evaluated the activity of β-galactosidase in cells treated with dacarbazine by substrate hydrolysis. Cell adhesion strength was measured by centrifugal assay application with subsequent staining of adhesive cells with Ki-67 monoclonal antibodies. Ability of melanoma cells to metabolize dacarbazine was determined by expressional analysis of CYP1A1, CYP1A2, CYP2E1 followed by CYP1A1 protein level evaluation by the ELISA method. The present study determined that dacarbazine treatment of melanoma cells could induce an increase in the percentage of cells in G0 phase without alterations of β-galactosidase positive cells which corresponded to the fraction of the senescent cells. Transcriptomic profiling of cells under dacarbazine induction of G0 -positive cells percentage revealed that 'VEGFA-VEGFR2 signaling pathway' and 'Cell cycle' signaling were mostly enriched by dysregulated genes. 'Focal adhesion' signaling was also found to be triggered by dacarbazine. In melanoma cells treated with dacarbazine, an increase in G0 -positive cells among adherent cells was found. Dacarbazine induces the alteration in a percentage of melanoma cells residing in G0 phase of a cell cycle. The altered adhesive phenotype of cancer cells under transition in the G0 phase may refer to a specific intercellular communication pattern of quiescent/senescent cancer cells.

A comprehensive analysis of penile cancer in the region with the highest worldwide incidence reveals new insights into the disease

BackgroundAlthough penile cancer (PC) is uncommon in developed countries, it is widespread in developing countries. The state of Maranhão (Northeast, Brazil) has the highest global incidence recorded for PC, and, despite its socioeconomic vulnerability, it has been attributed to human papillomavirus (HPV) infection. This study aimed to determine the histopathological features, the prevalence of HPV infection, and the immunohistochemical profile of PC in Maranhão.MethodsA retrospective cohort of 200 PC cases were evaluated. HPV detection was performed using nested-PCR followed by direct sequencing for genotyping. Immunohistochemistry (IHC) was performed using monoclonal antibodies anti-p16INK4a, p53, and ki-67.ResultsOur data revealed a delay of 17 months in diagnosis, a high rate of penile amputation (96.5%), and HPV infection (80.5%) in patients from Maranhão (Molecular detection). We demonstrated the high rate of HPV in PC also by histopathological and IHC analysis. Most patients presented koilocytosis (75.5%), which was associated with those reporting more than 10 different sexual partners during their lifetime (p = 0.001). IHC revealed frequent p16INK4a overexpression (26.0%) associated with basaloid (p < 0.001) and high-grade tumors (p = 0.008). Interestingly, p16 appears not to be a better prognostic factor in our disease-free survival analysis, as previously reported. We also demonstrated high ki-67 and p53 expression in a subset of cases, which was related to worse prognostic factors such as high-grade tumors, angiolymphatic and perineural invasion, and lymph node metastasis. We found a significant impact of high ki-67 (p = 0.002, log-rank) and p53 (p = 0.032, log-rank) expression on decreasing patients’ survival, as well as grade, pT, stage, pattern, and depth of invasion (p < 0.05, log-rank).ConclusionsOur data reaffirmed the high incidence of HPV infection in PC cases from Maranhão and offer new insights into potential factors that may contribute to the high PC incidence in the region. We highlighted the possible association of HPV with worse clinical prognosis factors, differently from what was observed in other regions. Furthermore, our IHC analysis reinforces p16, ki-67, and p53 expression as important diagnosis and/or prognosis biomarkers, potentially used in the clinical setting in emerging countries such as Brazil.

Abstract 5721: c-Myc expression coupled with loss of cytoplasmic Miz-1 expression and its re-localization to a nuclear expression in high grade cholangio and hepatocholangio carcinomas

Abstract Miz-1 (Myc-interacting zinc finger) activates target genes through transcription factors that are regulated by specific antimitogenic signals. Association with Myc converts Miz-1 from being a transcriptional activator to a repressor. This binding and the formation of Myc/Miz-1 complex induces a change in the biophysical properties of the Miz-1 protein, rendering the complex insoluble and causing to relocalize inside the nucleus. In addition, this association stabilizes Myc and as a result represses transcription. With uncontrolled tumor progress, metastatic tumor expansion can occur through epithelial-to-mesenchymal transition (EMT) and the induction of ZEB-1 (Zinc Finger E-Box Binding Homeobox-1). Zeb-1 expression is highly associated with aggressive behavior tumor types. A cohort of 98 human cholangiocarcinoma carcinoma (CC) including 14 with mixed (CC)/(HCC) resected cases whose formalin fixed paraffin embedded (FFPE) tissue sections containing adjacent benign and malignant tumor lesions were examined by immunohistochemistry (IHC) using Miz-1 (ZBTB17), ZEB-1 (OT13G6), c-Myc (Y69), and Ki-67 (Mib-1) monoclonal antibodies. The clinicopathologic variables, including viral hepatitis, cirrhosis, tumor size, grade and number, vascular invasion, tumor-node-metastasis (TNM) stage, were obtained from each patient’s medical records. The variables were assessed for correlation with the immunohistochemical results. Compared to the adjacent benign liver section, reduction of Miz-1 expression was observed in 98/98 (100%) (P&amp;lt;0.0001) cases, while positive expression of c-Myc, Ki-67 (&amp;gt; 5%), and ZEB-1 was observed in 18/98 (18.36%) (P&amp;lt;0.0001), 27/98 (27.55%) (P=0.000003), and 15/98 (15.30%) (P&amp;lt;0.00001) cases respectively. Of note, complete loss of Miz-1 expression was noted in 20/36 (55.55%) (P=0.106) high grade tumors (III/IV) associated with positive c-Myc, Ki-67 (&amp;gt;5%), ZEB-1, and nuclear Miz-1 expression in 12/20 (60%) (P=0.12) cases (4 CC and 8 mixed (CC)/(HCC) cases). All 12 cases with positive ZEB-1 had confirmed positive lymph node involvement. These results support the role of Miz-1 as one of the intermediary checkpoints required for the induction of cell cycle arrest and apoptosis. Positive c-Myc/ZEB-1 coupled with Miz-1 reduced cytoplasmic and its re-localization to a nuclear expression confirms the transcriptional repressor role of the Myc/Miz-1 complex and the activation of the ZEB-1 gene as a predictor of increased metastatic expansion, therapy resistance, and poor survival in high grade CC, and CC/HCC cases. These findings correlate with a cohort of an adequate number of differently graded lesions from 147 HCC cases that showed similar expression pattern of Miz-1, c-Myc and Zeb-1 to better confirm our hypothesis in epithelial derived human malignancies with frequently amplified 8q24 chromosome. Citation Format: Joeffrey J. Chahine, Sumeyye Culfaci, DongHyang Kwon, Pamela Tuma, Bhaskar V.S. Kallakury. c-Myc expression coupled with loss of cytoplasmic Miz-1 expression and its re-localization to a nuclear expression in high grade cholangio and hepatocholangio carcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5721.

Correlation of immunohistochemical expression of p16, Ki-67, and p53 with histopathological diagnosis of noninvasive cervical lesions: A multicenter study from South East Nigeria

Background: There is a high burden of cervical cancer in our environment. Most patients present late when the prognosis is guarded. Hence, accurate diagnosis of preinvasive lesions from cervical biopsies is important for clinical decisions and patient management. The aim of the study is to correlate the expression of p16INK4a, p53, and Ki-67 with histopathological diagnosis of noninvasive cervical lesions. Materials and Methods: The paraffin blocks of all cervical biopsies (excluding cases histologically diagnosed as invasive lesions) seen in two histopathology laboratories in Nnewi, Southeast Nigeria, over a 10-year period (2011–2020) were retrieved from the archives of both facilities. The cases were subjected to immunohistochemistry using p16INK4a, Ki-67, and p53 monoclonal antibodies. Results: There were 23 normal/reactive (45.1%), 6 low-grade squamous intraepithelial lesion (11.8%), and 22 hIL (43.1%). There is a very strong positive correlation between p16INK4a expression and the histopathological diagnosis (Spearman's correlation = 0.98). There is a strong positive correlation between Ki-67 expression and the histopathological diagnosis (Spearman's correlation = 0.70). There is a weak positive correlation between p53 expression and histopathological diagnosis (Spearman's correlation coefficient = 0.40). Conclusion: p16INK4a shows the best correlation with histopathological diagnosis of noninvasive cervical lesions and may be a very useful adjunct to H and E for diagnosing preinvasive cervical lesions. However, p53 correlates poorly with histopathologically diagnosed noninvasive cervical lesions and therefore may not be diagnostically useful.

Correlation of Somatostatin Receptor 1-5 Expression, [68Ga]Ga-DOTANOC, [18F]F-FDG PET/CT and Clinical Outcome in a Prospective Cohort of Pancreatic Neuroendocrine Neoplasms.

Simple SummaryThe need for prognostic and predictive biomarkers in pancreatic neuroendocrine neoplasms (PNENs) is great. Overexpression of somatostatin receptors (SSTRs) provides a molecular basis for imaging these tumors with 68Ga-labeled somatostatin (SST) PET/CT and for treatment with somatostatin analogs. We evaluated all 5 somatostatin receptors (SSTR1-5) with immunohistochemistry and prospectively compared the results with both [68Ga]Ga-DOTANOC and [18F]F-FDG PET/CT in a cohort of 21 non-functional (NF) PNENs. SSTR2 was the only SSTR subtype to correlate with [68Ga]Ga-DOTANOC PET/CT. High SSTR5 expression correlated with a low Ki-67 proliferation index, suggesting a better prognosis for these patients. Thus, our results confirm that SSTR2 has the highest impact on SSTR PET signaling of PNENs.Purpose: The aim of this study was to correlate immunohistochemical (IHC) tissue levels of SSTR1-5 with the receptor density generated from [68Ga]Ga-DOTANOC uptake in a prospective series of NF-PNENs. Methods: Twenty-one patients with a total of thirty-five NF-PNEN-lesions and twenty-one histologically confirmed lymph node metastases (LN+) were included in this prospective study. Twenty patients were operated on, and one underwent endoscopic ultrasonography and core-needle biopsy. PET/CT with both [68Ga]Ga-DOTANOC and [18F]F-FDG was performed on all patients. All histological samples were re-classified and IHC-stained with monoclonal SSTR1-5 antibodies and Ki-67 and correlated with [68Ga]Ga-DOTANOC and [18F]F-FDG PET/CT. Results: Expression of SSTR1-5 was detected in 74%, 91%, 80%, 14%, and 77% of NF-PNENs. There was a concordance of SSTR2 IHC with positive/negative [68Ga]Ga-DOTANOC finding (Spearman’s rho 0.382, p = 0.043). All [68Ga]Ga-DOTANOC-avid tumors expressed SSTR2 or SSTR3 or SSTR5. Expression of SSTR5 was higher in tumors with a low Ki-67 proliferation index (PI) (−0.353, 95% CI −0.654–0.039, p = 0.038). The mean Ki-67 PI for SSTR5 positive tumors was 2.44 (SD 2.56, CI 1.0–3.0) and 6.38 (SD 7.25, CI 2.25–8.75) for negative tumors. Conclusion: SSTR2 was the only SSTR subtype to correlate with [68Ga]Ga-DOTANOC PET/CT. Our prospective study confirms SSTR2 to be of the highest impact for SST PET/CT signal.

Postrecurrence Survival After Liver Transplantation for Liver Metastases From Neuroendocrine Tumors.

Liver metastases from neuroendocrine tumors (NETs) are an accepted indication for liver transplantation (LT). Despite strict patient selection, post-LT recurrence is observed in 30%-50% of cases. Postrecurrence survival is poorly investigated as well as factors influencing postrecurrence outcomes. Consecutive patients treated at a single institution for post-LT recurrence of NET between January 1, 2004, and December 31, 2018, were included. Baseline patients' characteristics, data on the primary tumor, pretransplant therapies, posttransplant recurrence and treatments, and long-term outcomes were prospectively collected and retrospectively analyzed. Thirty-two patients presented with post-LT NET recurrence occurring 82.9 mo (interquartile range, 29.4-119.1 mo) from LT, and the most common sites were abdominal lymph nodes (59.4%), peritoneum (6.3%), and lungs (6.3%). Fourteen patients (43.8%) underwent surgery with radical intent. Five- and 10-y survival after recurrence were 76.3% and 45.5%, respectively. Only time from LT to recurrence had a significant impact on postrecurrence survival, being 5-y overall survival 89.5% versus 0% for patients recurring >24 mo after LT versus ≤24 mo, respectively (P = 0.001). Moreover, for patients with Ki-67 monoclonal antibody staining >2% at recurrence, 5 y overall survival was 87.5% versus 0% for those undergoing surgery versus locoregional or systemic treatments (P = 0.011). The presented results, although based on a retrospective and relatively small series, show that excellent long-term survival is observed after post-LT NET recurrence, particularly in those patients recurring long after LT (>24 mo). An aggressive surgical treatment might result in a new chance of cure for a selected subgroup of patients.

Necroptosis Inhibition by Hydrogen Sulfide Alleviated Hypoxia-Induced Cardiac Fibroblasts Proliferation via Sirtuin 3

Myocardial ischemia or hypoxia can induce myocardial fibroblast proliferation and myocardial fibrosis. Hydrogen sulfide (H2S) is a gasotransmitter with multiple physiological functions. In our present study, primary cardiac fibroblasts were incubated with H2S donor sodium hydrosulfide (NaHS, 50 μM) for 4 h followed by hypoxia stimulation (containing 5% CO2 and 1% O2) for 4 h. Then, the preventive effects on cardiac fibroblast proliferation and the possible mechanisms were investigated. Our results showed that NaHS reduced the cardiac fibroblast number, decreased the hydroxyproline content; inhibited the EdU positive ratio; and down-regulated the expressions of α-smooth muscle actin (α-SMA), the antigen identified by monoclonal antibody Ki67 (Ki67), proliferating cell nuclear antigen (PCNA), collagen I, and collagen III, suggesting that hypoxia-induced cardiac fibroblasts proliferation was suppressed by NaHS. NaHS improved the mitochondrial membrane potential and attenuated oxidative stress, and inhibited dynamin-related protein 1 (DRP1), but enhanced optic atrophy protein 1 (OPA1) expression. NaHS down-regulated receptor interacting protein kinase 1 (RIPK1) and RIPK3 expression, suggesting that necroptosis was alleviated. NaHS increased the sirtuin 3 (SIRT3) expressions in hypoxia-induced cardiac fibroblasts. Moreover, after SIRT3 siRNA transfection, the inhibitory effects on cardiac fibroblast proliferation, oxidative stress, and necroptosis were weakened. In summary, necroptosis inhibition by exogenous H2S alleviated hypoxia-induced cardiac fibroblast proliferation via SIRT3.

Vitamin E ameliorates disturbances in testosterone pathway and sperm quality of male rats induced by the glycosides vicine and convicine of Vicia faba

PurposeThis paper aims to design to evaluate the protective effect of vitamin E to ameliorate the disturbances in testosterone pathway and sperm quality of male rats induced by the glycosides vicine (V) and convicine (C) of Vicia faba.Design/methodology/approachForty male albino rats were divided into five equal groups; control, paraffin oil, V (400 mg/kg) C (150 mg/kg)-treated group, vitamin E (100 mg/kg) + VC-treated group, and vitamin E (200 mg/kg) + VC-treated groups which injected intraperioneally (IP) with 0.5-ml saline, 0.5-ml paraffin oil,V (400 mg/kg) and C (150 mg/kg) of Vicia faba, vitamin E (100 mg/kg) + VC-treated groups, and Vitamin E(200 mg/kg) + VC-treated groups, respectively. Blood and testicular tissue were obtained after one month of the study. The male genital organs were calculated. Testosterone (Ts), luteinizing hormone (LH), follicle stimulating hormone (FSH), dehydroepiandrosterone sulfate (DHEA-SO4), sex hormone binding globulin (SHBG),?-glutamyl transpeptidase (?-GT), glucose-6-phosphate dehydrogenase (G6PD), 3ß-hydroxysteroid dehydrogenase (3ßHSD), lactate dehydrogenase (LDH), spermatozoa concentration, percent of mortality and abnormal sperms were evaluated.FindingsThe VC-treated group showed significant decrease (p < 0.01) in Ts, DHEA-SO4, G6PD, spermatozoa number and mortality percent, as well as, the male genital organs (testes, epidydemis, seminal vesicle, prostate and vasa deferentia) while significant increase (p < 0.01) was found in LH, FSH, SHBG, LDH, ?-GT, sperms monoclonal Ki-67, and abnormal spermatocytes levels compared with control group. Vitamin E co-injection with VC-treated group returned all these parameters to the normal values. The higher dose of vitamin E (200 mg/kg) was more effect than the lower dose (100 mg/kg).Originality/valueVicia faba contains V and C glycosides. The V and C glycosides in Vicia faba are hydrolyzed by intestinal microflora to aglycones divicine and isouramil, respectively. Divicine and isouramil are highly reactive compounds generating free radicals where divicine and isouramil are the main factors of favism. The V and C glycosides induced disturbances in testosterone pathway and sperm quality of male rats and vitamin E ameliorates these disturbances.

CD3/TCRE Expression and Immunoregulatory Milieu Induced in a Secondary Intermediate Host by Different Phases of Hydatid Cyst.

Echinococcosis is a common health problem in the Mediterranean and the Middle East, and manifests without any symptoms, even in the advanced stages. The present study aimed to investigate the cell mediated-immunoregulatory milieu in rats' echinococcosis induced by three different viability status of Echinococcus granulosus especially in the semi-calcareous stage, which can be used as novel biomarkers to monitor disease progression and open the door to a deeper understanding of the pathways that could contribute to complementary echinococcosis therapies. Rat infection with echinococcosis was induced by three different viable statuses of Echinococcus granulosus (G6) camel strain. During the different stages of parasitic infection, blood serum was harvested from rats containing low-, high-, and not viable (not completely transformed to the calcareous status) protoscoleces fluid. The host Th1/Th2 cytokines-mediated immune cell activation, as well as CD3/TCRE immunoregulation, and proliferation responses were investigated; especially in the semi-calcareous stage as this is the first report characterizing this stage. Both IFN-γ and IL-6 levels significantly increased in the infected groups (P < 0.05), in addition, increased positive immunoreactions in splenic tissue for both CD3/TCRE and Ki-67 monoclonal antibodies. E. granuloses infection-induced immune tolerance is involved in disease progression, and modulates the activation and regulation of host immune response, even in the early stages of infection, rather than the last stages of viability (semi-calcareous) is not neglected stage. This study is the first to report that the semi-calcareous stage causes a severe immunological response.

Determination and Comparison of Ki-67 Index in Astrocytic Tumours: A Tertiary Care Centre Experience

Introduction: The proper management of the astrocytic tumours largely depends on its correct diagnosis. But, smaller size and complicated histomorphology makes it difficult for the histopathologist to conclude the diagnosis. Monoclonal antibody Ki-67 {MIB-1/MIB-1 Labelling Index (LI)} plays a role of diagnostic as well as prognostic marker. It is an important marker, since it helps in deciding malignant potential of the astrocytic tumours where histology alone does not suffice. This study promotes the idea of including Ki-67 in routine practice for astrocytic tumours as it helps in quantifying the growth of the tumour which is of utmost importance in predicting the outcome accurately. Aim: The aim of this study was to determine the mean and ranges of Ki-67/MIB-1 LI in astrocytic tumours and comparing between different grades. Materials and Methods: This was a hospital based cross- sectional study with 50 cases of astrocytoma of varying grades over period of six years, retrospective study period started from 1st April 2009 till 30th November 2012 and prospective study period started from 1st December 2012 till 31st March 2014. Immunolabeling was done using Ki-67/MIB-1 antibody. The mean and ranges of Ki-67 was calculated in astrocytic tumours and its correlation with each World Health Organization (WHO) grade of histological diagnosis and clinical presentations were studied. One-way Analysis of Variance (ANOVA) and unpaired t-test were used for statistical analysis. Results: There were 50 astrocytic tumours, Pilocytic Astrocytoma (PA), grade I-5 (10%) cases; Diffuse Astrocytoma (DA), grade II-8 (16%) cases; Anaplastic Astrocytoma (AA), grade III-10 (20%) cases; and Glioblastoma Multiforme (GBM), grade IV-27 (54%) cases, with mean Ki-67 LI as 2.63%, 2.65%, 18.85% and 30.2%, respectively. The difference in mean Ki-67 LI for low grade astrocytoma and high grade astrocytoma was highly significant statistically (p&lt;0.0001). The most common presentation was seizure, which was seen in 26 (52%) cases. Conclusion: Histological grading with Ki-67 LI can work synergistically to reach the diagnosis, as both are subjected to heterogeneity induced diagnostic accuracy. Ki-67 LI increases with increase in the grade of the astrocytic tumours and hence, its quantification can help histopathologists as well as clinicians to agree on a point, especially in those cases where two differ diagnostically and it effects patients’ prognosis.

The features of proliferative processes in the thyroid gland of the Wistar rat’s offspring after intrauterine action of dexamethasone

The features of proliferative processes in the thyroid gland of the Wistar rat’s offspring after intrauterine action of dexamethasone were studied. Animals were divided into 3 groups: I – intact rats; II – control – animals, which on the 18th day of the dated pregnancy transuterine, transdermal, subcutaneously in the interscapular area was injected with 0.9 % saline in the amount of 0.05 ml; III – experimental group – animals, which during laparotomy by intrauterine, transdermal subcutaneous injection in the interscapular area was injected with a solution of dexamethasone at a dose of 0.05 ml at a dilution of 1:40 intrauterinely on the 18th day of pregnancy (Ukrainian patent No.112288). In the experimental subgroups used the allowable, generally accepted number of animals for statistical processing and obtaining reliable results – 6 animals. The thyroid gland with the tracheal area was removed on the 21st, 30th, 45th, 60th, 90th, 120th days of life. Immunohistochemical study was performed according to the protocol recommended for a particular antibody manufacturer. Monoclonal antibodies ki-67 (Ki-67), Fox-1 Antibody (A-12) were used to assess proliferative activity, the company Santa Cruz Biotechnology, Inc. (USA). The study found that the thyroid gland of rats of infantile period, which prenatally exposed to dexamethasone, is structurally represented by chaotically located follicles of different diameters with a predominance of large with desquamated cells in the lumen, and proliferative changes aimed at forming extrafollicular which is confirmed immunohistochemically by the presence of Ki-67-positive cells. Intracellularly, protein- synthesizing organelles of thyrocytes also proliferate, to which there is a clear cytoplasmic and nuclear reaction with Fox-1 antibodies. Dining the juvenile period, proliferative processes in the thyroid gland of animals of the experimental group are stabilized while maintaining the morphological structure of the hypofunctional type, and remain lower compared to the control and intact groups. Morphological signs of functional tension of the thyroid gland animals exposed prenatally to dexamethasone, which correlate with a decrease in proliferative activity, indicate a functional compensatory response of synthetic and hormone-producing function, but suppression of proliferative processes, despite the slight manifestations. The thyroid gland of morphological hypofunctional type after prenatal action of dexamethasone in young rats, indicates an adaptogenic compensatory response and morphofunctional immaturity of the organ during this period, which may be the basis for provoking the preservation of such morphogenetic factors under the influence of stressors. Keywords: thyroid gland, proliferation, dexamethasone, experiment, rats.

Orofacial skin inflammation increases the number of macrophages in the maxillary subregion of the rat trigeminal ganglion in a corticosteroid-reversible manner

Inflammation of the cutaneous orofacial tissue can lead to a prolonged alteration of neuronal and nonneuronal cellular functions in trigeminal nociceptive pathways. In this study, we investigated the effects of experimentally induced skin inflammation by dithranol (anthralin) on macrophage activation in the rat trigeminal ganglion. Tissue localization and protein expression levels of ionized calcium-binding adaptor molecule 1 (Iba1), a macrophage/microglia-specific marker, and proliferation/mitotic marker antigen identified by the monoclonal antibody Ki67 (Ki67), were quantitatively analyzed using immunohistochemistry and western blots in control, dithranol-treated, dithranol- and corticosteroid-treated, and corticosteroid-treated trigeminal ganglia. Chronic orofacial dithranol treatment elicited a strong pro-inflammatory effect in the ipsilateral trigeminal ganglion. Indeed, daily dithranol treatment of the orofacial skin for 3–5 days increased the number of macrophages and Iba1 protein expression in the maxillary subregion of the ipsilateral ganglion. In the affected ganglia, none of the Iba1-positive cells expressed Ki67. This absence of mitotically active cells suggested that the accumulation of macrophages in the ganglion was not the result of resident microglia proliferation but rather the extravasation of hematogenous monocytes from the periphery. Subsequently, when a 5-day-long anti-inflammatory corticosteroid therapy was employed on the previously dithranol-treated orofacial skin, Iba1 immunoreactivity was substantially reduced in the ipsilateral ganglion. Collectively, our findings indicate that both peripheral inflammation and subsequent anti-inflammatory therapy affect macrophage activity and thus interfere with the functioning of the affected sensory ganglion neurons.

An Immunohistochemical Study Showing Ki-67 as an Analytical Marker in Oral Malignant and Premalignant Lesions.

ABSTRACTIntroduction:Ki-67 is a nuclear protein. It is a proliferation marker that has an essential function in tumorigenesis due to its positive connection with tumor expansion.Aim:The aim of this study was to evaluate the articulation of Ki‑67 as prognostic marker in various grades of oral epithelial dysplasia (OED) and in oral squamous cell carcinoma (OSCC).Materials and Methods:A total of 100 histologically affirmed samples of normal oral mucosa (NOM), OED, and OSCC were divided into three groups—Group I (10 samples of normal oral mucosa), Group II (45 samples of OED), Group III (45 samples of OSCC). Routine hematoxylin and eosin and immunohistochemical staining with Ki-67 monoclonal antibody were carried out in all the samples.Results:Within Group I, articulation of Ki-67 was constrained to the basal layers. In Group II, cells showing positive expression of Ki-67 were available in the basal, suprabasal, and spinous layers. Cells showing positive expression of Ki-67 among well-differentiated OSCC were presented mainly in the periphery of the tumor nests; in moderately differentiated OSCC, cells were located in both peripheral and part of a center of the tumor nests; and in most cases of poorly differentiated OSCC, cells were diffused. Statistically significant difference in positive expression of Ki-67 was appreciated between three groups.Conclusion:Ki-67 antigen may perhaps be used as a marker for the histological reviewing of OED and OSCC. With the increase in the severity of OED, cells showing positive expression of Ki-67 also increased.

To the question of proliferative verrucous leukoplakia malignant potential

To study of neoplastic transformation of the epithelial cells of the oral mucosa with verrucous hyperortokeratosis, verrucous carcinoma and oral squamous cell carcinoma (OSCC). Oral mucous membrane biopsies of 33 patients with clinical diagnosis of proliferative verrucous leukoplakia were investigated. Histologically in 19 cases (57.6%) was revealed verrucous hyperorthokeratosis, in 8 cases (24.2%) - verrucous carcinoma and in 6 cases (18.2%) - oral squamous cell carcinoma. Tissue antigens were determined using mouse monoclonal antibodies to Ki-67 and mouse monoclonal antibodies to cytokeratin 15. In comparison with the proliferative activity of epithelial cells in the malpighian layer in verrucous hyperorthokeratosis (17.2±8.1%) was detected increasingof cell proliferation in epithelial growth zone in verrucous carcinoma (33.8±8.1%) and in peripheral zone of solid areas in oral squamous cell carcinoma (51.2±35.7%). A significant decrease in the expression of cytokeratin 15 in the cytoplasm of tumor cells was noted inverrucous carcinoma and oral squamous cell carcinoma in comparison with verrucous hyperorthokeratosis. The obtained results proclaim that there is no stage of epithelial dysplasia during tumor transformation of proliferative verrucous leukoplakia into verrucous carcinoma and oral squamous cell carcinoma. Verrucous carcinoma by the nature of proliferative activity, cell atypia and expression of cytokeratin 15 corresponds to carcinoma in situ.