Mast cells in oral lichen planus and oral lichenoid lesions related to dental amalgam contact

Abstract The aim of this study was to analyze the expression of mast cell markers toluidine blue, c-kit, and tryptase and presence of mononuclear inflammatory cells in oral lichen planus (OLP) and oral lichenoid lesions related to dental amalgam. Nineteen specimens of OLP, OLLC, and healthy oral mucosa were selected. Mononuclear inflammatory cells were analyzed. Histochemical and immunohistochemical analyses were performed using toluidine blue, anti-c-kit and anti-tryptase reagents, and the results were quantified in areas A and B of connective tissue. Mast cells of all OLP and OLLC samples were positive for toluidine blue, c-kit, and tryptase. The density of toluidine blue+, c-kit+ and tryptase+ mast cells was higher in tissue with OLP and OLLC compared with healthy controls (p < 0.05). No difference was noted in mast cells density between OLP and OLLC (p > 0.05). The density of tryptase+ mast cells was higher in the subepithelial region (area A) than the region below it (Area B) in OLLC (p = 0.047). The mononuclear inflammatory cell density was higher in OLLC compared to OLP, but without statistical significance (p > 0.05). A positive statistical correlation was found between mononuclear immune cells and density of c-kit+ and tryptase+ mast cells in OLP (r = 0.943 and r = 0.886, respectively). Our data demonstrate that the etiopathogenesis process of OLP and OLLC modulates the expansion and degranulation of mast cells; mast cells density, however, was similar between OLP and OLLC. The distribution of mast cells appears to vary along the lamina propria.

Mechanism of Bcl-2 BH4 selective inhibitor BDA-366 on NK/T cell lymphoma cells / 国际肿瘤学杂志

Objective:To investigate the inhibitory effect and killing mechanism of Bcl-2 BH4 selective inhibitor BDA-366 on NK/T cell lymphoma (NK/TCL) .Methods:Human NK cell leukemia cell line YT and human NK/TCL cell line NK92 cells were treated with 0, 0.05, 0.10, 0.20, 0.30, 0.40, 0.50 μmol/L BDA-366. CCK-8 assay was used to calculate the half inhibitory concentration (IC 50) value of BDA-366 on these cells. The apoptosis levels of cells in control group and IC 50 BDA-366 treated group were detected by flow cytometry. Western blotting was used to detect the expression levels of apoptosis-related proteins in cells of control group and 1/2 IC 50, IC 50, 2× IC 50 BDA-366 treated groups. TMRE and Fluo-3 fluorescent probe were used to detect mitochondrial membrane potential of control group and IC 50 BDA-366 treated group, and the intracellular Ca 2+ concentration of control group, IC 50, 2× IC 50 BDA-366 treated groups. NOD-SCID mice in control group and 10 mg/kg BDA-366 intraperitoneal injection group were weighed and HE staining was performed to evaluate the toxicity of BDA-366 in vivo. Results:The IC 50 of BDA-366 for YT and NK92 cells were 0.065 and 0.086 μmol/L respectively. The apoptosis rates of YT cells in the control group and 0.065 μmol/L BDA-366 group were (6.62±1.59) % and (34.60±3.06) % respectively. The apoptosis rates of NK92 cells in the control group and 0.086 μmol/L BDA-366 group were (5.57±0.88) % and (29.18±0.90) % respectively, both with statistically significant differences ( t=14.05, P<0.001; t=32.58, P<0.001). The relative expression of Bax in NK92 cells of the control group, 0.043, 0.086 and 0.172 μmol/L BDA-366 groups were 0.85±0.00, 1.26±0.04, 1.51±0.18, 1.15±0.10 ( F=20.70, P<0.001), the relative expression of Bax in BDA-366 groups were higher than that in the control group (all P<0.05). The fluorescence intensity of TMRE of YT cells in the control group and 0.065 μmol/L BDA-366 group were 8 372.00±330.47 and 6 419.67±311.34, and that of NK92 cells in the control group and 0.086 μmol/L BDA-366 group were 9 169.00±535.72 and 7 311.67±295.52 respectively, and there were statistically significant differences ( t=7.45, P=0.002; t=5.26, P=0.006). In YT cells, the intracellular Ca 2+ concentrations of 0.065 and 0.130 μmol/L BDA-366 groups were significantly higher than that of the control group (5 791.67±220.45, 6 729.33±585.39, 4 874.67±112.61, F=19.16, P=0.003) ( P=0.039; P=0.002). In NK92 cells, the intracellular Ca 2+ concentrations of 0.086 and 0.172 μmol/L BDA-366 groups were significantly higher than that of the control group (4 553.67±17.62, 4 740.33±254.50, 4 185.67±17.67, F=10.96, P=0.010) ( P=0.039; P=0.007). There was no statistically significant difference in body weight change on day 12 compared with day 0 of NOD-SCID mice between BDA-366 group and control group [ (3.18±0.01) g vs. (2.73±0.58) g, t=0.60, P=0.570], and HE staining showed no abnormal morphology of heart, liver, spleen, lung and kidney in BDA-366 group. Conclusion:BDA-366 promotes NK/TCL cells apoptosis in vitro, but does not cause weight loss and morphological changes of organs by HE staining in vivo. The inhibitory effect of BDA-366 on NK/TCL cells may be achieved by increasing Bax expression, inducing Ca 2+ release and reducing mitochondrial membrane potential.

Effect of astragaloside IV on PI3K/Akt signaling pathway in substantia nigra of mice with Parkinson′s disease / 中华麻醉学杂志

Objective:To evaluate the effect of astragaloside IV on phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in substantia nigra of mice with Parkinson′s disease.Methods:Forty-five SPF healthy male C57BL/6 mice, aged 8 weeks, weighing 19-25 g, were divided into 3 groups ( n=15 each) using a random number table method: control group (group C), Parkinson′s disease group (group PD) and astragaloside IV group (group A). The mouse model of Parkinson′s disease was developed by intraperitoneal injection of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) 30 mg/kg everyday for 7 consecutive days.Astragaloside 20 mg/kg was intraperitoneally injected everyday at 30 min before MPTP injection for 7 consecutive days before the model was prepared in group A, and the equal volume of normal saline was given instead in group C. Behavior was measured at 1 day interval after completion of administration.The mice were then sacrificed, and the substantia nigra of the brain tissue were obtained for determination of the expression of tyrosine hydroxylase(TH), phosphorylated PI3K(p-PI3K), phosphorylated Akt(p-Akt), glial fibrillary acidic protein (GFAP) and brain-derived neurotrophic factor (BDNF) (by Western blot). Results:Compared with C group, the total distance of movement and latency of falling were significantly shortened, the hanging score was decreased, the step width was increased, the expression of TH, p-PI3K, p-Akt and BDNF in substantia nigra was down-regulated, and the expression of GFAP was up-regulated in PD group and A group ( P<0.05). Compared with PD group, the total distance of movement and the latency to fall were significantly prolonged, the hanging score was increased, the step width was reduced, and the expression of TH, p-PI3K, p-Akt and BDNF in the substantia nigra was up-regulated, and the expression of GFAP was down-regulated in group A ( P<0.05). Conclusions:The mechanism by which astragaloside IV improves motor dysfunction is related to the activation of PI3K/Akt signaling pathway, up-regulation of BDNF expression and inhibition of astrocyte activation in mice with Parkinson′s disease.

Glucagon like peptide-1 mediated PI3K/Akt pathway antagonizes advanced oxidized protein products induced apoptosis of HTR-8/SVneo cells / 中国医师杂志

Objective:To explore the protective mechanism of phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) pathway in glucagon like peptide-1 (GLP-1) antagonizing the apoptosis of gestational trophoblasts (HTR-8/SVneo) induced by advanced oxidized protein products (AOPP).Methods:Pregnant trophoblast HTR-8/SVneo were cultured in vitro. The cells were divided into control group, AOPP group, GLP-1 group, AOPP + GLP-1 group and AOPP + GLP-1 + LY294002 group. The control group was cultured in 1640 medium; AOPP group was stimulated with 200 μg/ml AOPP; GLP-1 group was stimulated with 50-100 nmol/L GLP-1 for 1 h; AOPP + GLP-1 group was stimulated with 200 μg/ml AOPP for 48 hours, and then GLP-1 (50-100 nmol/L) was added for 1 hour; In AOPP + GLP-1 + LY294002 group, PI3K inhibitor LY294002 was added on the basis of the intervention of AOPP + GLP-1 group. The expression of PI3K/Akt pathway related protein p-Akt was detected by Western blot. Cell viability was detected by cell counting kit (CCK-8). Enzyme linked immunosorbent assay (ELISA) was used to detect the contents of apoptosis promoter protease caspase-9 and caspase-3, and the contents of apoptosis related proteins Bcl-2, Bax and Cyto-c. Results:After AOPP stimulation, the expression of p-Akt in AOPP group was lower than that in control group ( P<0.05); After 50 and 100 nmol/L GLP-1 intervention, the expression of p-Akt in AOPP + GLP-1 group was significantly higher than that in AOPP group (all P<0.05). After 24 and 48 hours of 100 nmol/L GLP-1 intervention, the expression of p-Akt in AOPP + GLP-1 group was significantly higher than that in AOPP group (all P<0.05). After AOPP stimulation, the cell viability of AOPP group was lower than that of control group ( P<0.05); After GLP-1 intervention, the cell viability of AOPP + GLP-1 group was significantly higher than that of AOPP group ( P<0.05). After adding PI3K inhibitor LY294002, the cell viability of AOPP + GLP-1 + LY294002 group was significantly lower than that of AOPP + GLP-1 group ( P<0.05). The results of ELISA showed that the contents of apoptosis promoter protein caspase-3, caspase-9, apoptosis related protein Bax and Cyto-c in AOPP group were higher than those in control group (all P<0.05), and the content of anti-apoptosis protein Bcl-2 was lower than that in control group ( P<0.05); After GLP-1 intervention, the contents of caspase-3, caspase-9, Bax and Cyto-c in AOPP + GLP-1 group were significantly lower than those in AOPP group ( P<0.05), and the content of anti-apoptosis protein Bcl-2 was higher than that in AOPP group ( P<0.05). After treatment with PI3K inhibitor LY294002, the contents of Bcl-2 in AOPP + GLP-1 + LY294002 group were lower than those in AOPP + GLP-1 group, and the contents of Bax and Cyto-c were higher than those in AOPP + GLP-1 group (all P<0.05). Conclusions:GLP-1 may mediate PI3K / Akt pathway to antagonize the apoptosis of HTR-8/SVneo induced by AOPP.

Effect of C-KIT gene mutation on the prognosis of acute myeloid leukemia associated with core-binding factor in children: a Meta-analysis / 白血病·淋巴瘤

Objective:To systematically evaluate the relationship between C-KIT gene mutation and the prognosis of childhood core-binding factor-related acute myeloid leukemia (CBF-AML).Methods:The PubMed database was searched with "KIT" "Acute Myeloid Leukemia" and "Children"; the Chinese Journal Full-text Database (CNKI), Chinese Biomedical Literature Database (CBM), VIP database and Wanfang database were also searched with "KIT" "Acute Myeloid Leukemia" and "Children", and the search time was from the establishment of the database to October 1, 2020. After strict screening, the literature was included in the analysis; according to the presence or absence of genetic changes, the included cases was divided into C-KIT mutation group and wild group, and the complete remission (CR) rate, event-free survival (EFS) rate, and overall survival (OS) rate of the two groups were analyzed.Results:Six articles were collected, including 4 articles in English and 2 articles in Chinese, with a total of 667 patients. There was a statistically significant difference in the EFS rate between the C-KIT mutation group and wild group in children with CBF-AML ( HR = 2.40, 95% CI 1.47-3.89, P = 0.001); there was no significant difference in the CR rate and OS rate ( OR = 0.93, 95% CI 0.48-1.80, P = 0.830; HR = 1.92, 95% CI 0.96-3.83, P = 0.065) between the two groups. Conclusions:C-KIT gene mutation may be a risk factor for poor prognosis in children with CBF-AML.

La vía canónica PI3K/AKT/mTOR y sus alteraciones en cáncer / The PI3K/AKT/mTOR canonical pathway and its alterations in cancer

RESUMEN La vía PI3K/AKT/mTOR participa en múltiples procesos celulares fundamentales para la célula. Algunas mutaciones genéticas de los componentes de esta vía se han asociado a diversas enfermedades humanas: las más importantes son los carcinomas de mama, tiroides y endometrio, el glioblastoma multiforme, el cáncer de próstata y los linfomas. La vía canónica PI3K/AKT/mTOR se ha estudiado ampliamente en los últimos años. Sin embargo, el conocimiento de la complejidad de sus componentes principales y su interrelación con los elementos de otras vías va en aumento. Por ello, es importantes actualizar cada cierto tiempo la información disponible para la comprensión de este mecanismo. Así mismo, se están y se han desarrollado numerosos ensayos con medicinas selectivas en búsqueda de un tratamiento más inteligente para las enfermedades asociadas a alteraciones de esta vía. Por tanto, realizamos una revisión de esta vía de transducción con el objetivo de tener una visión cercana de su funcionamiento, sus alteraciones y enumerar algunas moléculas promisorias para ser utilizadas en futuros tratamientos.

ABSTRACT The PI3K/AKT/mTOR pathway is involved in multiple cellular processes which are essential for the cells. Some genetic mutations of the components of this pathway have been associated with various human diseases, the most important of which are breast, thyroid and endometrium carcinomas; glioblastoma multiforme; prostate cancer and lymphomas. The PI3K/AKT/mTOR canonical pathway has been extensively studied in recent years. However, as the complexity of its main components and their correlation with the components of other pathways are increasing, it is important to update from time to time the available information to understand this mechanism. Furthermore, many trials have been conducted with selective medicines aimed to look for a more intelligent treatment for diseases associated with alterations in this pathway. Therefore, we review this transduction pathway to take a close look at its functioning and alterations, and to list some promising molecules for future treatments.

Role of heat-shock proteins in infection of human adenocarcinoma cell line MCF-7 by tumor-adapted rotavirus isolates / Papel de las proteínas de choque térmico en la infección de la línea celular de adenocarcinoma humano MCF-7 por aislados de rotavirus adaptados a tumores

Abstract Background: Viruses are being used as alternative and complementary tools for treating cancers. Oncolytic viruses exhibit tumor tropism, ability to enhance anti-tumor immunity and ability to be used in combination with conventional chemotherapy and radiotherapy. We have recently selected some rotavirus isolates which are adapted to efficiently infect and kill tumor cell lines. Aim: We tested five tumor cell-adapted rotavirus isolates for their ability to infect the human adenocarcinoma cell line MCF-7. Methods: Cell surface membrane-associated proteins mediating virus particle attachment were characterized using ELISA, immunoprecipitation, FACS analysis, and antibody blocking. Results: It was found that heat shock proteins (HSPs) such as Hsp90, Hsp70, Hsp60, and Hsp40 are expressed on the cell surface forming complexes with protein disulfide isomerase (PDI), integrin β3, and heat shock cognate protein 70 (Hsc70) in lipid raft microdomains. Interaction of rotavirus isolates with these cellular proteins was further confirmed by a competition assay and an inhibition assay involving the HSPs tested. Conclusion: Our findings suggest that the tumor cell-adapted rotavirus isolates studied here offer a promising tool for killing tumor cells, thus encouraging further research into this topic, including animal models.

Resumen Antecedentes: Los virus se utilizan como herramientas alternativas y complementarias para el tratamiento del cáncer. Los virus oncolíticos exhiben tropismo por tumores, capacidad para intensificar la inmunidad antitumoral y la capacidad para utilizarse en combinación con quimioterapia y radioterapia convencionales. Recientemente, hemos seleccionado algunos aislamientos de rotavirus que están adaptados para infectar y eliminar de manera eficiente líneas de células tumorales. Objetivo: Se ensayaron cinco aislamientos de rotavirus adaptados a células tumorales para determinar su capacidad para infectar la línea celular de adenocarcinoma humano MCF-7. Métodos: Las proteínas asociadas a la membrana de la superficie celular que median la unión de partículas de virus se caracterizaron mediante ELISA, inmunoprecipitación, análisis FACS y bloqueo de anticuerpos. Resultados: Se encontró que las proteínas de choque térmico (HSPs) como Hsp90, Hsp70, Hsp60 y Hsp40 se expresan en la superficie celular formando complejos con la proteína disulfuro isomerasa (PDI), la integrina β3 y la proteína análoga de choque térmico 70 (Hsc70) en microdominios lipídicos (rafts). La interacción de los aislamientos de rotavirus con estas proteínas celulares se confirmó adicionalmente mediante un ensayo de competición y un ensayo de inhibición que incluía las HSP ensayadas. Conclusión: Nuestros hallazgos sugieren que los aislamientos de rotavirus adaptados a las células tumorales estudiados aquí ofrecen una herramienta prometedora para eliminar las células tumorales, lo que estimula más investigaciones sobre este tema, incluidos los modelos animales.

Are stem cell marker expression and cd133 analysis relevant to differentiate colorectal cancer? / A expressão de marcadores de células-tronco e a análise do cd133 são relevantes na diferenciação do câncer colorretal?

ABSTRACT Background: CD133 and AXL have been described as cancer stem cell markers, and c-MYC as a key regulatory cellular mechanism in colorectal cancer (CRC). Aim: Evaluate the prognostic role of the biomarkers CD133, AXL and c-MYC and their association with clinicopathologic characteristics in colorectal adenocarcinomas and adenomas. Methods: A total of 156 patients with UICC stage I-IV adenocarcinomas (n=122) and adenomas (n=34) were analyzed. Tissue microarrays (TMA) from primary tumors and polyps for CD133, c-MYC and AXL expression were performed and analyzed for their significance with clinicopathologic characteristics. Results: Poorly differentiated adenocarcinomas and disease progression were independent risk factors for poor overall survival. The median overall survival time was 30 months. Positive CD133 expression (35.9% of all cases), particularly of right-sided CRCs (44.8% of the CD133+ cases), was negatively correlated with death in the univariate analysis, which did not reach significance in the multivariate analysis. c-MYC (15.4% of all cases) was predominantly expressed in advanced-stage patients with distant (non-pulmonary/non-hepatic) metastasis. AXL expression was found only occasionally, and predominantly dominated in adenomas, with less penetrance in high-grade dysplasia. Conclusions: CD133 expression was not associated with inferior overall survival in CRC. While AXL showed inconclusive results, c-MYC expression in primary CRCs was associated with distant metastasis.

RESUMO Racional: CD133 e AXL são descritos na literatura como marcadores de células-tronco tumorais, e c-MYC cumpre papel chave como mecanismo de regulação celular no câncer colorretal (CCR). Objetivo: Avaliar o papel prognóstico dos biomarcadores CD133, AXL e c-MYC e sua associação com características clinicopatológicas de adenocarcinomas e adenomas colorretais. Métodos: Um total de 156 pacientes com adenocarcinomas de estádio UICC I-IV (n=122) e adenomas (n=34) colorretais foram avaliados. Microarranjos teciduais (TMA) dos tumores primários e adenomas foram realizados em busca de expressão de CD133, c-MYC e AXL, com posterior análise de relação significativa com características clinicopatológicas. Resultados: Adenocarcinomas pobremente diferenciados e progressão de doença foram fatores de risco independentes para má sobrevida global. A taxa mediana de sobrevida global foi de 30 meses. Expressão positiva de CD133 (35,9% dos casos), particularmente em cânceres de cólon direito (44,8% dos casos CD133+), correlacionou-se negativamente com óbito na análise univariada, sem significância estatística na análise multivariada. c-MYC (15,4% dos casos) teve predomínio de expressão em pacientes com estádio avançado com metástases distantes (não-pulmonares/não-hepáticas). Expressão de AXL foi pouco encontrada, com predomínio em adenomas, com menor penetrância em displasia de alto grau. Conclusão: Expressão de CD133 não se associou com sobrevida global inferior em CCR. Enquanto AXL demonstrou resultados inconclusivos, expressão de c-MYC em tumores primários se associou-se à metástases à distância.

MiR-26a promotes angiogenesis in rats with cerebral ischemia through PI3K/Akt pathway / 国际脑血管病杂志

Objective:To investigate the effect of miR-26a mediated phosphatidylinositol-3 kinase (PI3K)/protein kinase B (AKT) signaling pathway on angiogenesis in rats with cerebral ischemia.Methods:A total of 100 male SD rats were divided into sham operation group, model group, miR-NC group, and miR-26a group according to the random number table method. The miR-NC group and the miR-26a group were injected with 5 μl miR-26a simulant negative control and miR-26a simulant into the lateral ventricle respectively. The sham operation group and the model group were injected with the same amount of normal saline respectively. The middle cerebral artery occlusion model was induced by the modified intraluminal suture method. In the sham operation group, the thread was only inserted without ligation. Five rats in each group were injected intraperitoneally with 5-bromodeoxyuridine (BrdU) daily for 7 days. Rat brain microvascular endothelial cells (BMECs) cultured and transfected in vitro were divided into control group, oxygen glucose deprivation (OGD) group, miR-NC group, and miR-26a group. The dual luciferase experiment verified the regulatory effect of miR-26a on the phosphatase and tensin homolog deleted on chromosome 10 (PTEN). Longa score was used to detecte the neurological damage of rats. The volume of cerebral infarction was measured by triphenyltetrazolium chloride (TTC) staining. The methyl thiazolyl tetrazolium (MTT) staining, annexin Ⅴ fluorescein isothiocyanate/propidium iodide double staining and tubule formation experiment were used to detect the proliferation, apoptosis and angiogenesis of BMECs, respectively. Real-time fluorescence quantitative reverse transcription polymerase chain reaction was used to detect the miR-26a expression of ischemic brain tissue and BMECs. Immunofluorescence double labeling method (BrdU/von Willebrand factor [vWF]) was used to detect the proliferation of rat vascular endothelial cells. Western blot analysis was used to detect the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), angiopoietin-2 (Ang-2), PTEN, PI3K and Akt protein in ischemic brain tissue.Results:Bioinformatics and dual luciferase experiments verified the targeted regulation of PTEN by miR-26a. Compared with the sham operation group, the expression of miR-26a, VEGF, bFGF, Ang-2, PI3K, AKT and the number of BrdU + /VWF + cells in ischemic brain tissue in the model group and miR-NC group increased, while the expression of PTEN decreased (all P<0.05). Compared with the model group, the effect of various indexes in the miR-26a group was more significant (all P<0.05). Compared with the control group, the proliferation and angiogenesis of BMECs in the OGD group and the miR-NC group were significantly increased, and the apoptosis was significantly reduced (all P<0.05). Compared with the OGD group, the effect of various indexes in the miR-26a group was more significant (all P<0.05). Conclusion:miR-26a can mediate the targeted inhibition of PTEN expression, up-regulate angiogenesis related factors (VEGF, bFGF and Ang-2), and promote vascular endothelial cell proliferation and angiogenesis in rats with cerebral infarction by activating PI3K/Akt signaling pathway.

Role of SPP1 in endogenous protective mechanism underlying neuropathic pain in mice with spinal cord injury: relationship with VEGF-AKT signaling pathway / 中华麻醉学杂志

Objective:To evaluate the role of secreted phosphoprotein 1 (SPP1) in endogenous protective mechanism underlying neuropathic pain (NP) in mice with spinal cord injury and the relationship with the vascular endothelial growth factor (VEGF)/kinase B (Akt) signaling pathway.Methods:Seventy-two clean-grade healthy female Kunming mice, aged 7-8 weeks, weighing 30-35 g, were divided into 4 groups ( n=18 each) using a random number table method: sham group (Sham group), NP caused by spinal cord injury group (NP group), NP caused by spinal cord injury+ SPP1-siRNA group (NS-siRNA group), and NP caused by spinal cord injury+ adeno-associated virus vector group (NP-EV group). A model of NP was established by a semi-transecting of the spinal cord.AAV-SPP1-siRNA-GFP adenovirus and AAV-vector-GFP adenovirus 7 μl were intrathecally injected in NS-siRNA group and NP-EV group, respectively, and 5 days later the model was established.At 1, 2 and 3 weeks after operation, 6 mice in each group were randomly selected to measure paw withdrawal threshold to mechanical stimulation (PWMT) and tail flick latency (TFL) to thermal stimuli.And then the mice were sacrificed and the ipsilateral injured spinal cord tissues were taken for determination of the expression of SPP1 mRNA (by real-time polymerase chain reaction) and expression of SPP1, VEGF, Akt and phosphorylated Akt (p-Akt) (by Western blot). Results:Compared with group Sham, PWMT was significantly decreased, TFL was shortened, and the expression of SPP1 mRNA, SPP1, VEGF and p-Akt protein was up-regulated at 1, 2 and 3 weeks after operation in NP, NS-siRNA and NP-EV groups( P<0.05). Compared with group NP, PWMT was significantly decreased, TFL was shortened, and the expression of SPP1 mRNA, SPP1, VEGF and p-Akt protein was down-regulated at 1, 2 and 3 weeks after operation in group NS-siRNA( P<0.05). Compared with group NS-siRNA, PWMT was significantly increased, TFL was prolonged, and the expression of SPP1 mRNA, SPP1, VEGF and p-Akt protein was up-regulated at 1, 2 and 3 weeks after operation in group NS-siRNA( P<0.05). Conclusion:SPP1 is involved in the endogenous protective mechanism underlying NP in mice with spinal cord injury, which may be related to the activation of the VEGF/AKT signaling pathway.

Correlation of c-MET and CXCR4 proteins and microvessel density with liver metastasis in colorectal cancer / 肿瘤研究与临床

Objective:To explore the correlation of c-MET and CXCR4 proteins and microvessel density (MVD) with liver metastasis in colorectal cancer tissues.Methods:A total of 40 colorectal cancer tissue samples and 10 paracancerous (5 cm from the edge of the tumor) normal colorectal tissue samples were collected from March 2015 to December 2020 in Shanxi Traditional Chinese Medical Hospital. Among 40 patients with colorectal cancer, 15 patients had liver metastasis. Immunohistochemistry was used to detect c-MET protein, CXCR4 protein and CD34-labeled MVD in various tissues, and the relationships between them and liver metastasis and between the three were analyzed.Results:The positive rates of c-MET protein [72.5% (29/40) vs. 30.0% (3/10)], CXCR4 protein [47.5% (19/40) vs. 10.0% (1/10)] and MVD (20.1±5.2 vs. 11.5±4.3) in colorectal cancer tissues were higher than those in paracancerous tissues, and the differences were statistically significant (all P < 0.05). The positive rates of c-MET protein [86.7% (13/15) vs. 64.0% (16/25)] and CXCR4 protein [66.7% (10/15) vs. 36.0% (9/25)] in colorectal cancer liver metastasis group were significantly higher than those in non-liver metastasis group, and the differences were statistically significant (both P < 0.05). MVD in colorectal cancer liver metastasis group was significantly higher than that in non-liver metastasis group (21.5±5.3 vs. 12.4±5.7), and the difference was statistically significant ( P < 0.05). In colorectal cancer tissues, c-MET protein expression was positively correlated with CXCR4 protein expression ( r = 0.568, P < 0.05), and MVD in c-MET-positive patients or CXCR4-positive patients was higher than that in negative ones (both P < 0.05). Conclusions:The c-MET protein, CXCR4 protein and MVD may play important roles in the liver metastasis of colorectal cancer. The three indicators can provide a certain reference for the early diagnosis and prognosis prediction of liver metastasis of colorectal cancer.

Are stem cell marker expression and cd133 analysis relevant to differentiate colorectal cancer? / A expressão de marcadores de células-tronco e a análise do cd133 são relevantes na diferenciação do câncer colorretal?

ABSTRACT Background: CD133 and AXL have been described as cancer stem cell markers, and c-MYC as a key regulatory cellular mechanism in colorectal cancer (CRC). Aim: Evaluate the prognostic role of the biomarkers CD133, AXL and c-MYC and their association with clinicopathologic characteristics in colorectal adenocarcinomas and adenomas. Methods: A total of 156 patients with UICC stage I-IV adenocarcinomas (n=122) and adenomas (n=34) were analyzed. Tissue microarrays (TMA) from primary tumors and polyps for CD133, c-MYC and AXL expression were performed and analyzed for their significance with clinicopathologic characteristics. Results: Poorly differentiated adenocarcinomas and disease progression were independent risk factors for poor overall survival. The median overall survival time was 30 months. Positive CD133 expression (35.9% of all cases), particularly of right-sided CRCs (44.8% of the CD133+ cases), was negatively correlated with death in the univariate analysis, which did not reach significance in the multivariate analysis. c-MYC (15.4% of all cases) was predominantly expressed in advanced-stage patients with distant (non-pulmonary/non-hepatic) metastasis. AXL expression was found only occasionally, and predominantly dominated in adenomas, with less penetrance in high-grade dysplasia. Conclusions: CD133 expression was not associated with inferior overall survival in CRC. While AXL showed inconclusive results, c-MYC expression in primary CRCs was associated with distant metastasis.

RESUMO Racional: CD133 e AXL são descritos na literatura como marcadores de células-tronco tumorais, e c-MYC cumpre papel chave como mecanismo de regulação celular no câncer colorretal (CCR). Objetivo: Avaliar o papel prognóstico dos biomarcadores CD133, AXL e c-MYC e sua associação com características clinicopatológicas de adenocarcinomas e adenomas colorretais. Métodos: Um total de 156 pacientes com adenocarcinomas de estádio UICC I-IV (n=122) e adenomas (n=34) colorretais foram avaliados. Microarranjos teciduais (TMA) dos tumores primários e adenomas foram realizados em busca de expressão de CD133, c-MYC e AXL, com posterior análise de relação significativa com características clinicopatológicas. Resultados: Adenocarcinomas pobremente diferenciados e progressão de doença foram fatores de risco independentes para má sobrevida global. A taxa mediana de sobrevida global foi de 30 meses. Expressão positiva de CD133 (35,9% dos casos), particularmente em cânceres de cólon direito (44,8% dos casos CD133+), correlacionou-se negativamente com óbito na análise univariada, sem significância estatística na análise multivariada. c-MYC (15,4% dos casos) teve predomínio de expressão em pacientes com estádio avançado com metástases distantes (não-pulmonares/não-hepáticas). Expressão de AXL foi pouco encontrada, com predomínio em adenomas, com menor penetrância em displasia de alto grau. Conclusão: Expressão de CD133 não se associou com sobrevida global inferior em CCR. Enquanto AXL demonstrou resultados inconclusivos, expressão de c-MYC em tumores primários se associou-se à metástases à distância.

Neutral lactate promotes proliferation, migration and invasion of hepatocellular carcinoma cells by activating pi3k/akt signaling pathway / 肿瘤

Objective: To investigate the effects of neutral lactate on the proliferation, migration and invasion of hepatocellular carcinoma (HCC) cells, and to explore its possible molecular mechanism. Methods: The HCC SMMC-7721 and HepG2 cells were treated with 0 (as the control), 2.5, 5 and 10 mmol/L neutral lactate, respectively. Then the proliferation of HCC cells was detected by MTT assay. The migration and invasion abilities of HCC cells were detected by Transwell chamber method. The expression levels of phosphoinositide 3-kinase (PI3K), protein kinase B (PBK, Akt), phospho-Akt (Ser473) [p-Akt (S473)], matrix metallopeptidase-2 (MMP-2) and MMP-9 were detected by Western blotting. The HCC cells SMMC-7721 and HepG2 were divided into four groups, and treated with equal volume of medium (as the control), neutral lactate, perifosine (an inhibitor of PI3K/Akt pathway), and neutral lactate combined with perifosine, respectively. Then MTT assay and Transwell chamber assay were used to detect the proliferation, migration and invasion abilities of HCC cells in each group. The expression levels of PI3K, Akt, p-Akt (S473), MMP-2 and MMP-9 proteins in HCC cells were detected by Western blotting. Results: The proliferation, migration and invasion abilities of SMMC-7721 and HepG2 cells in 5 and 10 mmol/L neutral lactate treatment groups were significantly enhanced as compared with the control group (all P < 0.01). The expression levels of PI3K, p-Akt (S473), MMP-2 and MMP-9 proteins in HCC cells treated with neutral lactate were significantly increased as compared with the control group (all P < 0.01). Compared with the control group, the proliferation, migration and invasion abilites of HCC cells in the perifosine treatment group were significantly decreased (all P < 0.05), and the expression levels of PI3K, p-Akt (S473), MMP-2 and MMP-9 proteins were also significantly down-regulated (all P < 0.05). Compared with the neutral lactate treatment group, the proliferation, migration and invasion abilities of HCC cells in the neutral lactate combined with perifosine treatment group were significantly reduced (all P < 0.01), and the expression levels of PI3K, p-Akt (S473), MMP-2 and MMP-9 proteins were significantly decreased (all P < 0.01). Conclusion: Neutral lactate promotes the proliferation, migration and invasion of HCC cells by activating PI3K/Akt pathway, while the PI3K/ Akt inhibitor perifosine can significantly reduce the stimulative effects of neutral lactate on the proliferation, migration and invasion of HCC cells.

AKT modulates PTEN expression in hepatocarcinoma HepG2 cells via C-MYC pathway / 肿瘤

Objective: To investigate whether C-myc is involved in the modulation of protein kinase B (PKB, also known as AKT) on the expression of phosphate and tension homology deleted on chromsome ten (PTEN) in hepatoma HepG2 cells. Methods: HepG2 cells were treated with different concentrations of AKT inhibitor Capivasertib (5, 10 and 20 nmol/L). The expression levels of AKT, phosphorylated (p)-AKT, Bad, p-Bad, C-myc and p-C-myc proteins were detected by Western blotting. The transcription levels of C-myc downstream genes eukaryotic initiation factor-4E (eIF-4E), branched‐chain amino acid aminotransferase 1 (BCAT1) and WW domain-containing E3 ubiquitin protein ligase 1 (WWP1) were detected by real-time fluorescent quantitative PCR. The expressions of eIF-4E, BCAT1 and WWP1 genes were silenced by siRNA, then the transcription and expression of PTEN were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The effects of Capivasertib treatment or WWP1 gene silence on the expression and localization of PTEN protein in HepG2 cells were detected by cell immunofluorescence technique. Results: In HepG2 cells treated with different concentrations of Capivasertib for 8 h, the expression levels of p-AKT, p-C-myc and p-Bad were significantly downregulated as compared with the control group (all P < 0.01), and the expression levels of eIF-4E, BCAT1 and WWP1 mRNAs were also downregulated (all P < 0.01). Silencing eIF-4E, BCAT1 and WWP1 gene expressions had no significant effect on the transcription of PTEN (all P 0.01) but the silence of WWP1 gene could significantly enhance the expression of PTEN protein (P < 0.01). Both Capivasertib treatment and WWP1 gene silence could increase the expression level of PTEN and induce its aggregation on the cell membrane. Conclusion: AKT is able to affect the transcription of WWP 1 gene by C-myc pathwaty, and ultimately participates in the modulation of PTEN expression in HepG2 cells.

Clinicopathological features of dedifferentiated liposarcomas with meningothelial-like whorls: report of six cases / 中华病理学杂志

Objective@#To investigate the histopathologic, immunohistochemical, molecular genetic characteristics of dedifferentiated liposarcomas with meningothelial-like whorls(DDLMW).@*Methods@#Six cases of DDLMW diagnosed at Jiangsu Province Hospital(the First Affiliated Hospital of Nanjing Medical University) from March 2012 to August 2018 were enrolled. The cases were analyzed by routine HE staining, immunohistochemistry(MDM2, CDK4 and p16) and fluorescent in-situ hybridization(FISH) on MDM2 gene. Related literatures were also reviewed.@*Results@#Three of the 6 patients were male.The patient ages ranged from 40 to 77 years (mean, 58 years). Four tumors occurred in the retroperitoneum and two in the mediastinum. Histologically, the tumors showed, in addition to foci of well-differentied liposarcoma, characteristic, scattered meningothelial-like concentrical whorls. The whorls were composed of tightly, concentrically arranged, spindle to ovoid cells with mild to mederate cytological atypia. Metaplastic bone was present within or in their immediate vicinity in four case. The tumors cell also showed strong and diffuse immunoreactivity to MDM2, CDK4 and p16, but no immunoreactivity to S-100 protein, SMA, SOX10, EMA, CD21, CD23 or CD35. The Ki-67 labeling indexes were low, while FISH showed high levels of MDM2 amplification in all cases.@*Conclusions@#DDLMW is a rare morphologic variant of dedifferentiated liposarcoma. The whorls in DDLMW do not represent perineurial or follicular dendritic differentiation. Recognition and familiarity with its existence, as well as combined application of immunohistochemical staining and MDM FISH, are important to avoid confusion with other lesions.

Clinicopathological features of dedifferentiated liposarcomas with meningothelial-like whorls: report of six cases / 中华病理学杂志

To investigate the histopathologic, immunohistochemical, molecular genetic characteristics of dedifferentiated liposarcomas with meningothelial-like whorls(DDLMW). Six cases of DDLMW diagnosed at Jiangsu Province Hospital(the First Affiliated Hospital of Nanjing Medical University) from March 2012 to August 2018 were enrolled. The cases were analyzed by routine HE staining, immunohistochemistry(MDM2, CDK4 and p16) and fluorescent in-situ hybridization(FISH) on MDM2 gene. Related literatures were also reviewed. Three of the 6 patients were male.The patient ages ranged from 40 to 77 years (mean, 58 years). Four tumors occurred in the retroperitoneum and two in the mediastinum. Histologically, the tumors showed, in addition to foci of well-differentied liposarcoma, characteristic, scattered meningothelial-like concentrical whorls. The whorls were composed of tightly, concentrically arranged, spindle to ovoid cells with mild to mederate cytological atypia. Metaplastic bone was present within or in their immediate vicinity in four case. The tumors cell also showed strong and diffuse immunoreactivity to MDM2, CDK4 and p16, but no immunoreactivity to S-100 protein, SMA, SOX10, EMA, CD21, CD23 or CD35. The Ki-67 labeling indexes were low, while FISH showed high levels of MDM2 amplification in all cases. DDLMW is a rare morphologic variant of dedifferentiated liposarcoma. The whorls in DDLMW do not represent perineurial or follicular dendritic differentiation. Recognition and familiarity with its existence, as well as combined application of immunohistochemical staining and MDM FISH, are important to avoid confusion with other lesions.

Immunophenotypic characteristics of diffuse large b-cell lymphoma and expression of C-MYC protein: single Ecuadorian center experience / Características inmunofenotípicas del linfoma difuso de células grandes B y expresión de la proteína C-MYC: experiencia en un centro de Ecuador

Abstract Background: Diffuse large B-cell lymphoma (DLBCL) makes up from 25% to 40% of all non-Hodgkin lymphomas (NHL) and is the most common histological subtype worldwide. In Ecuador, DLBCL makes up 49% of all NHL cases, but there have been no studies on the immunophenotypic classificationof DLBCL in germinal center (GC) and non-germinal center (NGC)subtypes.This study was conducted to ascertain the immunophenotypic profile of DLBCL in an Ecuadorian hospital. Methods: A total of 38 DLBCL cases from 2006 to 2015 were compiled from the Pathology Service at Metropolitan Hospital (HM) in Quito, Ecuador. Eleven of these cases failed to meet the inclusion criteria; thus, the final sample consisted of 27 cases. Manual tissue microarrays were constructed, and three immunohistochemical markers (CD10, BCL6, and MUM1) were applied according to the Hans algorithm; in addition, the expression of the c-myc protein expression was also investigated. Results: The results showed that 77.8% of cases were of the GC subtype, 11.1% were NGC, and 11.1% were unclassifiable according to the Hans algorithm. Conclusions: The most frequent DLBCL subtype was GC, with 21 cases; and 40.7% of these cases overexpressed c-myc.

Resumen Antecedentes: El linfoma difuso de células grandes B (LDCGB) constituye el 25 al 40% del total de los linfomas no Hodgkin (LNH) y es el subtipo histológico más frecuente en el mundo. En Ecuador el LDCGB corresponde al 49% del total de los casos de LNH, sin embargo no hay estudios de clasificación inmunofenotípica del LDCGB en centro germinal (CG) y no centro germinal (NCG). Este estudio se realizó para conocer el perfil inmunofenotípico del LDCGB en un hospital de Ecuador. Métodos: Se recopiló del Servicio de Patología del Hospital Metropolitano de Quito, Ecuador, un total de 38 casos de LDCGB desde el 2006 al 2015, de los cuales 11 no cumplieron con los criterios de inclusión. La muestra final fue de 27 casos. Se realizaron microarreglos tisulares manuales para la aplicación de tres marcadores de inmunohistoquímica según el algoritmo de Hans (CD10, BCL6 y MUM1) y luego se correlacionó con la sobreexpresión de la proteína c-MYC. Resultados: El 77,8% de casos fue tipo CG, 11,1% fue NCG y 11,1% fueron inclasificables según Hans. Conclusiones: El subtipo de LDCGB más frecuente fue CG con 21 casos y de estos 40,7% sobreexpresaron c-MYC.

Primary colonic liposarcomatosis: report of a case with review of literature

The colon is a rare site of occurrence of liposarcoma, as either the primary site or by secondary involvement from a retroperitoneal liposarcoma. Liposarcomatosis denotes simultaneous occurrence of multiple liposarcomas. There are only 17 cases of primary colonic liposarcoma reported in the English literature­one of which was primary colonic liposarcomatosis. We depict the second case of primary colonic liposarcomatosis in a 57-year-old female who presented with abdominal swelling and pain. On exploratory laparotomy, two large masses were seen arising from the wall of the right colon along with multiple smaller masses attached to the colon. Right hemicolectomy with en bloc excision of the masses was performed along with hysterectomy and pelvic floor repair. Macroscopically, multiple exophytic masses and one endophytic mass were identified. The exophytic masses were of variable size and were found to hang from the colon by a thin pedicle simulating variable-sized appendices epiploicae. Histopathologically, the lesions showed the morphology of well-differentiated liposarcoma. This appears to be a case of primary colonic liposarcomatosis. There is only one other similar case reported in the English literature, to the best of our knowledge.

Linfoma tipo Burkitt con afectación ovárica como única manifestación inicial: caso clínico y revisión de la literatura / Ovarian Burkitt lymphoma as the primary manifestation: a case report and literature review

La afectación ovárica como debut de un linfoma de Burkitt sin enfermedad extraovárica detectable es anecdótica, por lo que habitualmente no se incluye como hipótesis diagnóstica tras el hallazgo de una tumoración ovárica. Su desconocimiento lleva a realizar un tratamiento equivocado que puede llegar a comprometer el deseo reproductivo de la paciente. Presentamos el caso de una paciente que presenta un linfoma de Burkitt con afectación ovárica como manifestación inicial. La paciente desarrolló una progresión sistemática excepcionalmente rápida. A propósito de este caso y de su inusual evolución, revisamos la literatura existente.

Ovarian involvement as the initial manifestation of a Burkitt lymphoma without detectable extra-ovarian disease is rare, which is why it is usually not included in the differential diagnosis when an ovarian tumor is detected. A missed diagnosis will lead to the wrong treatment being given, and this can compromise any future reproductive wishes of the patient. In this article, a patient presents a Burkitt lymphoma with ovarian involvement as an initial manifestation and an unusually rapid systemic progression of the disease. Prompted by this case and its unusual course, we reviewed the existing literature.

Expression and significance of phosphorylated protein kinase B and spleen tyrosine protein kinase in different TNM stages of gastric cancer patients / 中国医师杂志

Objective To investigate the expression of phosphorylated protein kinase B (PKB/AKT) and spleen tyrosine protein kinase (Syk) in different tumor node metastasis (TNM) stages of gastric cancer patients.Methods From January 2015 to April 2018,82 patients with gastric cancer confirmed by gastroscopy and surgical pathology were enrolled in this study.All patients were selected for cancer tissue,and 30 patients were randomly selected from normal gastric mucosa at 5 cm adjacent to the tumor.Immunohistochemistry was used to detect the expression of PKB/AKT and Syk protein.To compare the expression of PKB/AKT and Syk in gastric cancer and adjacent normal tissues,and to analyze the relationship between PKB/AKT and Syk expression and clinicopathological features in gastric cancer tissues,and the correlation between PKB/AKT and Syk and TNM staging of gastric cancer patients.Results The positive expression rate of PKB/AKT in gastric cancer tissues was higher than that in adjacent tissues (P ＜ 0.05).The positive expression rate of Syk was lower than that in adjacent tissues (P ＜ 0.05).PKB/Akt and Syk gene expression in gastric cancer were related to histological grade,tumor infiltration,TNM staging,lymph node metastasis and distant metastasis (P ＜ 0.05),and the expression of PKB/AKT was significantly increased with the increase of TNM staging in gastric cancer patients,and the positive expression of Syk was significantly decreased (P ＜ 0.05).Conclusions PKB/AKT is positively correlated with TNM staging of gastric cancer patients.Syk is negatively correlated with TNM staging of gastric cancer patients.The clinical expression of PKB/AKT and Syk can be used to determine the TNM staging of gastric cancer,which provides a strong basis for tumor treatment.It is of great significance in treatment.