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Background: Lung cancer is the most common cause of cancer mortality worldwide. Smoking is undoubtedly the major risk factor of lung cancer in both genders. Adenocarcinoma is the most common form of lung cancer in both men and women and the most prevalent subtype in non-smokers. Lung cancer in never-smokers is a distinct entity with sparse studies. We studied the clinico-pathologic profile of lung adenocarcinoma and pattern of p53 expression in smokers and non-smokers. Methods: A prospective study involving 100 lung adenocarcinoma cases from January 2020 to June 2021 examined p53 expression using immunohistochemistry. Trucut biopsies, fine needle aspiration cytology (FNAC) cell blocks, and pleural effusion were analyzed to identify the predominant morphological subtype of the lung adenocarcinoma. Results: The most common histological pattern of lung adenocarcinoma was solid, and the presenting symptoms were cough and dyspnoea in both smokers and non-smokers. The incidence of lung adenocarcinoma was higher in non-smokers in the study. p53 expression had a significant correlation with smoking but not with stage of disease or morphological subtype of lung adenocarcinoma. Conclusions: p53 mutation has a statistical correlation with smoking in adenocarcinomas in our population. Among the adenocarcinoma cases in our study, non-smokers predominate (n=53). Even though our study showed the p53 mutation has no statistical correlation with the stage of the disease or histological subtype in adenocarcinoma, more cases need to be studied to prove this observation.
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SUMMARY: Although tacrolimus (TAC) significantly reduces allograft rejection incidence in solid-organ transplantation, its long-term use is associated with an increased risk of TAC-induced nephrotoxicity. In this study, we investigated the renoprotective effects of green tea extract (GTE) with or without the dipeptidyl peptidase 4 inhibitor, gemigliptin, by assessing serum creatinine levels, the amount of proteinuria, and histopathology in TAC-induced nephrotoxicity. TAC-induced nephrotoxicity was induced by intraperitoneal TAC injection, GTE was administered via subcutaneous injection, and gemigliptin was administered orally. Mice with TAC-induced nephrotoxicity exhibited a significant increase in both serum creatinine levels and 24-hour urine protein. However, when treated with GTE via subcutaneous injection, mice showed a decrease in serum creatinine levels and the amount of proteinuria. When GTE was combined with gemigliptin, further renoprotective effects were observed in biochemical assessments, consistent with the attenuation of TAC-induced nephrotoxicity in histopathology. The expression of p53 protein was lower in the mice treated with the combination of GTE and gemigliptin compared to mice with TAC-induced nephrotoxicity. Our results demonstrate that the combination of GTE and gemigliptin treatment reveals synergistic renoprotective effects by decreasing the expression of p53 protein. These findings suggest that the combination of GTE and gemigliptin could potentially be used as a prophylactic or therapeutic strategy for TAC-induced nephrotoxicity.
Aunque tacrolimus (TAC) reduce significativamente la incidencia de rechazo de aloinjertos en trasplantes de órganos sólidos, su uso a largo plazo se asocia con un mayor riesgo de nefrotoxicidad inducida por TAC. En este estudio, investigamos los efectos renoprotectores del extracto de té verde (GTE) con o sin el inhibidor de la dipeptidil peptidasa 4, gemigliptina, mediante la evaluación de los niveles de creatinina sérica, la cantidad de proteinuria y la histopatología en la nefrotoxicidad inducida por TAC. La nefrotoxicidad inducida por TAC se indujo mediante inyección intraperitoneal de TAC, el GTE se administró mediante inyección subcutánea y la gemigliptina se administró por vía oral. Los ratones con nefrotoxicidad inducida por TAC mostraron un aumento significativo tanto en los niveles de creatinina sérica como en la proteína en orina de 24 horas. Sin embargo, cuando se trataron con GTE mediante inyección subcutánea, los ratones mostraron una disminución en los niveles de creatinina sérica y en la cantidad de proteinuria. Cuando se combinó GTE con gemigliptina, se observaron efectos renoprotectores adicionales en las evaluaciones bioquímicas, lo que concuerda con la atenuación de la nefrotoxicidad inducida por TAC en histopatología. La expresión de la proteína p53 fue menor en los ratones tratados con la combinación de GTE y gemigliptina en comparación con los ratones con nefrotoxicidad inducida por TAC. Nuestros resultados demuestran que la combinación de tratamiento con GTE y gemigliptina revela efectos renoprotectores sinérgicos al disminuir la expresión de la proteína p53. Estos hallazgos sugieren que la combinación de GTE y gemigliptina podría usarse potencialmente como estrategia profiláctica o terapéutica para la nefrotoxicidad inducida por TAC.
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Animals , Mice , Piperidones/administration & dosage , Pyrimidines/administration & dosage , Tea , Plant Extracts/administration & dosage , Tacrolimus/toxicity , Kidney Diseases/drug therapy , Piperidones/pharmacology , Pyrimidines/pharmacology , Plant Extracts/pharmacology , Protective Agents , Drug Synergism , Immunosuppressive Agents/toxicity , Kidney/drug effects , Kidney Diseases/chemically inducedABSTRACT
Introducción: El cáncer de endometrio ocupa el sexto lugar en incidencia del cáncer en mujeres. La caracterización molecular de este cáncer permite optimizar la estratificación de riesgo para mejorar el tratamiento de las pacientes. Objetivo: Determinar el perfil molecular TCGA de pacientes con cáncer de endometrio en Bogotá, D.C., Colombia. Método: Estudio descriptivo en una cohorte de pacientes con cáncer de endometrio. Las mutaciones en los exones 9 a 14 del gen POLE fueron identificadas mediante amplificación por reacción en cadena de la polimerasa, seguida de secuenciación Sanger y análisis bioinformático. La expresión de las proteínas MMR y p53 se identificó mediante inmunohistoquímica. Resultados: Se incluyeron 40 pacientes con una mediana de edad de 66 años. El 15% presentaron mutaciones en el dominio exonucleasa de POLE. El 32% de las pacientes que no presentaron mutaciones manifestaron deficiencia en el sistema MMR. El 43,47% de las pacientes sin mutaciones en POLE ni alteración del sistema MMR presentaron alteración de la proteína p53. Conclusiones: La población de cáncer de endometrio analizada presenta un perfil molecular TCGA similar a lo reportado para otras poblaciones.
Introduction: Endometrial cancer ranks sixth in cancer incidence among women. Its molecular characterization allows for a more precise risk stratification with the aim of improving patient treatment. Objective: To determine the TCGA molecular profile of patients with endometrial cancer in Bogota, Colombia. Method: A descriptive study of a cohort of patients with endometrial cancer. The expression of MMR proteins and p53 was identified through immunohistochemistry. Mutations in exons 9 to 14 of the POLE gene were identified through polymerase chain reaction amplification, followed by Sanger sequencing and bioinformatic analysis. Results: Forty patients were included in the study, with a median age of 66 years, 15% of them exhibited mutations in the exonuclease domain of POLE, while 32% of patients without mutations showed deficiency in the MMR system. Forty three percent of patients without mutations in POLE or MMR alterations showed aberrant p53 protein expression. Conclusions: The analyzed population of endometrial cancer presents a TCGA molecular profile similar to that reported for other populations.
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Humans , Female , Middle Aged , Aged , Endometrial Neoplasms/genetics , Immunohistochemistry , Polymerase Chain Reaction , Cross-Sectional Studies , Retrospective Studies , Genes, p53/genetics , Endometrial Neoplasms/diagnosis , Endometrial Neoplasms/pathology , Sequence Analysis, DNA , Colombia , Risk Assessment , DNA Polymerase II , DNA Mismatch Repair , Poly-ADP-Ribose Binding Proteins , MutationABSTRACT
The clinical application of 5-fluorouracil (5-Fu), a potent chemotherapeutic agent, is often hindered by its well-documented cardiotoxic effects. Nevertheless, natural polyphenolic compounds like resveratrol (RES), known for their dual anti-tumor and cardioprotective properties, are potential adjunct therapeutic agents. In this investigation, we examined the combined utilization of RES and 5-Fu for the inhibition of gastric cancer using both in vitro and in vivo models, as well as their combined impact on cardiac cytotoxicity. Our study revealed that the co-administration of RES and 5-Fu effectively suppressed MFC cell viability, migration, and invasion, while also reducing tumor weight and volume. Mechanistically, the combined treatment prompted p53-mediated apoptosis and autophagy, leading to a considerable anti-tumor effect. Notably, RES mitigated the heightened oxidative stress induced by 5-Fu in cardiomyocytes, suppressed p53 and Bax expression, and elevated Bcl-2 levels. This favorable influence enhanced primary cardiomyocyte viability, decreased apoptosis and autophagy, and mitigated 5-Fu-induced cardiotoxicity. In summary, our findings suggested that RES holds promise as an adjunct therapy to enhance the efficacy of gastric cancer treatment in combination with 5-Fu, while simultaneously mitigating cardiotoxicity.
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Background Arsenic is a human carcinogen. Arsenic and its metabolites affect the expression of p53, but whether there are any changes of p53 phosphorylation and ubiquitination levels in human bronchial epithelium cells (BEAS-2B) are not clear after exposure to arsenic and its metabolites. Objective To study the effects of arsenic and its metabolites monomethylarsic acid (MMA) and dimethylarsinic acid (DMA) on the expression of tumor suppressor gene p53 in BEAS-2B cells. Methods Different concentrations of sodium arsenite (NaAsO2) were used to infect BEAS-2B cells, and the cell viability was detected with CCK-8 reagent to determine the dose and time of NaAsO2 used for the following study. Based on the results of cell viability, the cells were divided into two panels: a sodium arsenide panel and an arsenic methylation metabolite penal. The doses of sodium arsenite were 0, 2, 4, and 6 μmol·L−1 NaAsO2; the arsenic methylation metabolite panel consisted of 0 μmol·L−1 NaAsO2 group (control), 6 μmol· L−1 MMA group, 6 μmol· L−1 DMA group, and 6 μmol· L−1 NaAsO2 group. The cells were collected after 48 h treatment, and the total protein and total RNA were extracted. The relative levels of p53 mRNA expression were determined by quantitative real-time polymerase chain reaction (qRT-PCR), the relative expression levels of p53 protein, p53 Ser9 and Ser15 phosphorylated proteins were determined by Western blot, and the level of p53 ubiquitination was detected by co-immunoprecipitation (CO-IP). Results Compared with the control group, the cell viability rates in all BEAS-2B cells treated by NaAsO2 were significantly reduced (P<0.05), and the 50% cell viability was observed at 6 μmol·L−1. Compared with the control group, the relative expression level of p53 mRNA gradually decreased after NaAsO2 (2, 4, 6 μmol·L−1) treatment (P<0.05), the relative expression levels of p53 protein and Ser9 phosphorylated protein induced by NaAsO2 also decreased gradually (P<0.05), and the relative expression level of p53 Ser15 phosphorylated protein induced by NaAsO2 followed the same pattern, but it was only lower than that of the control group in the 6 μmol·L−1 NaAsO2 group (P<0.05). Compared with the control group, there were no significant effects on the relative expression levels of p53 mRNA, p53 protein, Ser9 and Ser15 phosphorylated proteins in the MMA group and the DMA group. Compared with the control group, the expression level of p53 ubiquitination was significantly decreased and the expression of K48 ubiquitination decreased significantly after NaAsO2 infection. Conclusion Arsenic causes a decrease in the expression of the p53 protein in BEAS-2B cells, largely due to inhibition of the phosphorylated pathway and a decrease in mRNA expression, and protein changes caused by a decrease in p53 ubiquitination do not play a dominant role. MMA and DMA do not affect p53 gene expression.
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Objective To explore the clinical significance and mechanisms of chromatin licensing and DNA repli-cation factor 1(CDT1)in lung adenocarcinoma).Methods The gene expression samples of lung adenocarcinoma tissue and normal lung tissue were downloaded from the TCGA database,and perform differential analysis,GO a-nalysis,independent prognosis analysis,and correlation analysis with immunotherapy using R language.CDT1 ex-pression in lung adenocarcinoma and normal tissues was detected by PCR in clinical samples.The changes of cell proliferation and cycle in si-CDT1 knockdown group and si-NC control group were detected by flow cytometry.The invasive ability of each group was detected by Transwell.The expressions of CDT1,TPX2 and p53 in each group were detected by Western blot.Results The TCGA data analysis revealed CDT1 as a differentially expressed gene.GO analysis indicated that CDT1 was closely associated with the cell cycle.The high expression of CDT1 in lung adenocarcinoma tissues was validated in clinical samples.CDT1 could serve as an independent factor for predicting the prognosis of lung adenocarcinoma and had predictive value for immunotherapy in lung adenocarcinoma.Knock-down of CDT1 resulted in a significant decrease in cell proliferation ability compared to the control group,and cells were noticeably arrested in the G1 phase.Transwell assay results demonstrated a significant reduction in invasive capacity in the CDT1 knockdown group.Knockdown of CDT1 led to a significant decrease in TPX2 expression and a significant increase in p53 expression,while overexpression of CDT1 yielded the opposite effect.Conclusion Re-sults demonstrate the elevated expression of CDT1 in lung adenocarcinoma,its association with prognostic signifi-cance,and its impact on lung adenocarcinoma's occurrence and development by influencing TPX2 and p53.
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Objective To investigate the clinical diagnostic value of plasma serine protease inhibitor Ka-zal-type 4(SPINK4)expression in colorectal adenocarcinoma(CRC)and progressive adenoma(AA).Methods A total of 62 patients with CRC(CRC group)and 15 patients with AA(AA group)diagnosed by colonoscopy and pathological examination in this hospital from June 2020 to December 2021 were selected,and 22 healthy people undergoing physical examination during the same period were selected as the HC group.The expression of SPINK4 in plasma was detected by ELISA,and the expression of CEA in plasma was detected by electrochemiluminescence,and the correlation was analyzed.The diagnostic efficiency was analyzed by re-ceiver operating characteristic(ROC)curve,and the expression of p53 in CRC tissues was detected by immu-nohistochemistry.Results The expression of plasma SPINK4 in the CRC group and AA group was lower than that in the HC group(Z=3.72,-0.41,P<0.05),and the expression of CEA in the CRC group was higher than that in the HC group(Z=-3.63,P<0.05).The area under the curve(AUC),accuracy,sensi-tivity and specificity of SPINK4 combined with CEA in the diagnosis of CRC and AA were higher than those of SPINK4 and CEA alone.The positive rate of mutant type p53 in SPINK4 low expression group and CEA high ex-pression group was significantly increased in CRC patients(72.55%,75.00%,P<0.05).Conclusion The expression of plasma SPINK4 is decreased in CRC and AA,and the combined detection of SPINK4 and CEA has a good di-agnostic efficiency in CRC and AA.
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Objective To investigate the efeects of microRNA(miR)-103a-3p regulates tumor protein 53-regulated inhibitor of apoptosis 1(TRIAP1)on osteoblast differentiation and bone mass in ovariectomized mice.Methods MC3T3-E1 cells were divided into normal group,miR-103a-3p-NC group,miR-103a-3p mimic group,miR-103a-3p mimic+TRIAP1-NC group,miR-103a-3p mimic+TRIAP1 mimic group.mRNA expression of miR-103a-3p,TRIAP1,P53 were detected by Real-time PCR;Cell proliferation and apoptosis were detected by MTT test and flow cytometry;cytoskeleton and mineralization of cells were detected by F-actin immunofluorescence staining and alizarin staining;alkaline phosphatase(ALP)activity was detected by ELISA.24 female mice were divided into sham group,osteoporosis(OP)group,miR-103a-3p antagonist-NC group,miR-103a-3p antagonist group(six in each group),extract bilateral ovaries to establish an OP model,sham group mice only isolated fat around ovarian tissue.mRNA expression of miR-103a-3p,TRIAP1,P53,ALP,osteocalcin(OCN),osteopontin(OPN)of bone tissue were detected;microCT detect bone mineral density(BMD),bone mineral content(BMC);haematoxylin eosin staining was used to observe pathological changes of bone tissue.Results After miR-103a-3p mimic was transfected into cells,the miR-103a-3p and P53 expression increased,TRIAP1 expression decreased,cell proliferation decreased,apoptosis increased,F-actin expression decreased,the number of calcium nodules decreased,and ALP enzyme activity decreased(P<0.01);however,after TRIAP1 mimic was additionally transfected into cells,the above result caused by miR-103a-3p mimics were significantly reversed(P<0.01).In OP group,the miR-103a-3p and P53 expression in bone tissue increased,the TRIAP1,ALP,OCN and OPN expression decreased,BMD and BMC were decreased,and bone tissue construct was damaged(P<0.05);in miR-103a-3p antagonist group,the miR-103a-3p and P53 expression in bone tissue decreased,TRIAP1,ALP,OCN,OPN expression increased,BMD and BMC increased,and bone tissue construct was improved(P<0.05).Conclusion MiRNA-103a-3p mediate TRIAP1/P53 to inhibit proliferation and mineralization of osteoblast,while miR-103a-3p antagonistic treatment reduce bone loss in OP mice.
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Objective:To explore the mechanism of zoledronic acid (ZOL) affects osteogenic differentiation and bone formation through the regulation of sirtuin 3 (SIRT3) / P53 expression.Methods:Bone marrow mesenchymal stem cells (BMSCs) were induced to differentiate into osteogenic cells, the expression of SIRT3 in the cells was detected, and the targeting regulation relationship between SIRT3 and P53 was analyzed. The intracellular expressions of SIRT3 and P53 were intervened and ZOL was used to treat the cells. MTT method, Western blot method and kit were used to detect cell viability, osteogenesis-related genes Osteoprotegerin (OPG), runt-related transcription factor 2 (Runx2) expression, alkaline phosphatase (ALP) activity and alizarin red S (ARS) staining, respectively. Ovariectomy (OVX) was used to construct a rat model and explore the effect of ZOL on the progression of osteoporosis (OP) in vivo.Results:ZOL promoted osteogenic differentiation of BMSCs. The expression of SIRT3 was down-regulated in the serum of OP patients (0.78±0.23) compared with that of healthy subjects (1.00±0.26 vs. 0.78±0.23. t=3.85, P<0.001). During the osteogenic differentiation of BMSCs, the expression level of SIRT3 gradually increased with the prolonged induction of osteogenesis. Compared with the p53 protein expression and BMSCs activity in the control group, SIRT3 knockout could increase the expression level of p53 protein (0.59±0.05 vs. 1.01±0.11. t=6.02, P=0.004) but inhibited the activity of BMSCs (100.00±8.41 vs. 51.26±5.59. t=8.36, P=0.001). After ZOL treatment, the inhibitory effect of SIRT3 on cell viability (49.61±5.11 vs. 87.61±7.31. t=7.38, P=0.002) and osteogenesis was relieved, and the level of P53 was inhibited (1.10±0.10 vs. 0.69±0.04. t=6.59, P=0.003). P53 overexpression partially offseted the effects of ZOL on cell viability (84.61±6.52 vs. 66.54±5.47. t=3.68, P=0.021) and osteogenesis. Compared with the sham surgery group, the OVX group showed inhibition of osteogenesis in rats, and ZOL treatment significantly improved osteogenic inhibition. ZOL treatment increased the expression level of SIRT3 protein in bone tissue of OVX rats, but inhibited the expression level of P53. Conclusion:ZOL promoted osteogenic differentiation and bone formation of BMSCs by promoting the ubiquitination and degradation of P53 by SIRT3.
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BACKGROUND:At present,effective preventive and therapeutic measures for hypertrophic scar are still limited.In contrast,most of botanical herbs have few side effects and abundant sources,offering new ideas and approaches for the prevention and treatment for hypertrophic scar. OBJECTIVE:To explore the potential molecular mechanism of plant-derived β-sitosterol on hypertrophic scar fibroblasts by network pharmacology and molecular docking techniques and to initially verify it by cytological experiments. METHODS:Through the network pharmacology,the relevant database and software were used to screen the drug targets of β-sitosterol and obtain the hypertrophic scar-related disease targets.The potential(intersection)targets of β-sitosterol on hypertrophic scar were obtained.Cytoscape software and STRING database were used to construct the"drug-target-disease"network and protein-protein interaction network,and screen out the core targets in the protein-protein interaction network.Gene ontology(GO)biological function and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses of intersection targets were conducted through the DAVID database,and the signaling pathways and core target genes closely related to the intersection targets were further identified through literature analysis.AutoDock software was used to perform the molecular docking of β-sitosterol and core target proteins.In vitro cellular assays were used to verify the effects of β-sitosterol on proliferation,apoptosis,cell cycle distribution and mRNA expression of core target genes in human hypertrophic scar fibroblasts. RESULTS AND CONCLUSION:There were 56 intersection targets of β-sitosterol and hypertrophic scar and 10 core targets were identified in the protein-protein interaction network,including tyrosine kinase,mitogen-activated protein kinase 3(MAPK3),cysteine protease 3(CASP3),apolipoprotein E,estrogen receptor 1,sterol regulatory element-binding transcription factor 1,peroxisome proliferator-activated receptor alpha,C-reactive protein,intercellular adhesion molecule 1,and catalase.Combined with the literatures and the functional analysis of the KEGG and GO,the MAPK signaling pathway was further identified to be closely related to the intersection targets,and MAPK3(ERK1-MAPK),CASP3,P53 and tumor necrosis factor were identified as the core targets.The molecular docking results indicated that β-sitosterol was well bound to the core target proteins.Cellular assays showed that 100 μmol/L β-sitosterol inhibited hypertrophic scar fibroblast proliferation,decreased mitochondrial membrane potential and induced apoptosis(P<0.01),increased the proportion of G1-phase cells and decreased the proportion of S-phase cells(P<0.05),upregulated the mRNA expression of CASP3,P53 and tumor necrosis factor(P<0.05),and downregulated the mRNA expression of MAPK3(P<0.001).To conclude,β-sitosterol may induce cell apoptosis in hypertrophic scar fibroblasts by activating the tumor necrosis factor pathway and upregulating the expression of CASP3 and P53,while inhibiting the ERK-MAPK pathway to arrest cell cycle and thus reduce the proliferation of hypertrophic scar fibroblasts.
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BACKGROUND:There are many treatment methods for knee osteoarthritis,among which electroacupuncture,as an important non-drug treatment,is effective in the treatment of knee osteoarthritis,but its exact mechanism is not clear. OBJECTIVE:Effect of electroacupuncture on the expression of p53 and P21 in articular cartilage and subchondral bone of aged rats with knee osteoarthritis. METHODS:Eight 6-month-old male Sprague-Dawley rats were included in the young group and sixteen 24-month-old male Sprague-Dawley rats were randomly divided into old group(n=8)and electroacupuncture group(n=8).The rats in the electroacupuncture group received electroacupuncture stimulation once a day,5 days a week,for 8 continuous weeks,and the other two groups did not do any treatment.Eight weeks later,the level of type Ⅱ collagen C-terminal peptide in peripheral blood was detected by ELISA,the morphology of left knee cartilage and subchondral bone was observed by safranin O-fast green staining,the degree of knee cartilage degeneration was evaluated by modified Mankin's score,the microstructure of left knee cartilage and subchondral bone was detected by micro-CT,and the expression levels of matrix metalloproteinase 13,P53,P21 Mrna and protein were detected by RT-PCR and western blot respectively. RESULTS AND CONCLUSION:Compared with the young group,the level of C-terminal peptide of type Ⅱ collagen in the peripheral blood was increased in the old group(P<0.05).The micro-CT results showed that the bone volume fraction,bone mineral density and the number of bone trabeculae were decreased in the old group compared with the young group(P<0.05),while the trabecular separation increased(P<0.05).Safranin O-fast green staining showed that in the old group,the surface layer of cartilage was uneven with fissures,the morphology of chondrocytes was irregular and stained unevenly,the boundary between the cartilage and subchondral bone was blurred,and the matrix loss was serious.The Mankin's score was higher in the old group than the young group(P<0.05).The expression of matrix metalloproteinase 13,P53,P21 at Mrna and protein levels increased in the old group compared with the young group(P<0.05).Compared with the old group,electroacupuncture decreased the level of C-terminal peptide of type Ⅱ collagen(P<0.05),increased the bone volume fraction,bone mineral density and the number of bone trabeculae(P<0.05),and decreased the trabecular separation(P<0.05).Safranin O-fast green staining showed that in the electroacupuncture group,the surface of cartilage was smooth and red staining was uniform,and the cell morphology and structure were between the young group and the old group.Following electroacupuncture treatment,the Mankin's score(P<0.05),matrix metalloproteinase 13 and P21 Mrna expression(P<0.05),and matrix metalloproteinase 13 and P53 protein expression decreased(P<0.05),while there was a decreasing trend of P53 Mrna and P21 protein expression,but with no statistical significance(P>0.05).To conclude,electroacupuncture may delay articular cartilage degeneration and subchondral osteoporosis in aged rats by inhibiting the expression of P53 and P21,so as to protect joints and delay joint aging.
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BACKGROUND:The clinical manifestation of acute radiation skin injury is recurrent necrotic ulcers,and its pathogenesis is still not fully understood.The establishment of a suitable animal model will have important clinical implications for the study of its pathogenesis,prevention and treatment. OBJECTIVE:To establish a model of acute β-ray radiation skin injury and to investigate the mechanism of injury. METHODS:Sixty-nine Sprague-Dawley rats were randomly divided into 30,45,60 Gy 32P-β-ray groups(n=21 per group)and control group(n=6).A single local irradiation of the back of the rats was performed using 32P radionuclide.The control group was operated in the same way as the irradiated groups except that it was not irradiated.The body mass and skin appearance of the rats were measured at 7,15,30,45,and 60 days after irradiation.Three rats from each group were selected at each observation time point.The skin injury was observed by hematoxylin-eosin staining,Masson staining,transmission electron microscopy,and TUNEL assay.P53,Bcl-2 and Bax protein levels in the skin were measured by immunohistochemistry and western blot assay. RESULTS AND CONCLUSION:There was no accidental death after irradiation,and the body mass of rats showed a gradual increase.The rats showed different degrees of epidermal necrosis,inflammatory cell infiltration,reduction of hair follicles and appendages,and collagen fibrillation,which were evident at 60 and 45 Gy.The levels of serum inflammatory factors,interleukin-6 and tumor necrosis factor-α,were significantly increased in a dose-dependent manner.Under the electron microscope,there are varying degrees of mitochondrial reduction,vacuolization and nuclear pyknosis in the cells.The degree of cell apoptosis showed a certain dose-dependence.Immunohistochemistry and western blot results showed an increase in the expression of P53 and Bax proteins and a decrease in the expression of Bcl-2 protein in the skin after irradiation.There were significant differences between the 60 Gy group and the 45 Gy and 30 Gy groups(P<0.05).To conclude,irradiation with 60 Gy and 45 Gy 32P radionuclide on the back of rats could successfully establish a practically pre-clinical animal model,and the mechanism is related to the up-regulation of P53 and Bax and the down-regulation of Bcl-2.This model can provide a reference for the establishment of animal models for the study of the mechanism of radiation skin injury and its prevention and treatment.
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BACKGROUND:Pulsed electromagnetic fields,as an important physical therapy,are exactly effective in the treatment of osteoarthritis,but the mechanism has not been fully clarified. OBJECTIVE:To observe the effect of pulsed electromagnetic field on the degeneration of knee joint cartilage in aged rats. METHODS:Eight 6-month-old Sprague-Dawley rats were selected as the young group and were subjected to normal diet with no treatment.Sixteen 22-month-old Sprague-Dawley rats were randomly divided into old group(n=8)and pulsed electromagnetic field group(n=8).The rats in the pulsed electromagnetic field group were subjected to a pulsed electromagnetic field intervention,once a day,5 days per week for continuous 8 weeks.The rats in the old group were given no treatment.All rats were anesthetized and executed after 8 weeks for the detection of relevant indexes. RESULTS AND CONCLUSION:Compared with the young group,serum type Ⅱ collagen C-terminal peptide level was increased in the old group(P<0.05);compared with the old group,serum type Ⅱ collagen C-terminal peptide level was decreased in the pulsed electromagnetic field group(P<0.05).Micro-CT showed that the bone volume fraction,bone mineral density,and number of bone trabeculae decreased(P<0.05)and the trabecular separation increased(P<0.05)in the tibia of rats in the aged group compared with the young group;and the bone volume fraction,bone density,and number of trabeculae increased(P<0.05)and the trabecular separation decreased(P<0.05)in the tibia of rats in the pulsed electromagnetic field group compared with the aged group.The tibial plateau Safranin O-fast green staining showed that the articular cartilage structure of rats in the aged group was disorganized,and the number of chondrocytes was obviously reduced,and the tidal line could not be distinguished.The above results were improved in the pulsed electromagnetic field group.RT-qPCR and western blot assay showed that the mRNA and protein expression levels of matrix metalloproteinase 1,matrix metalloproteinase 13,P53 and P21 in the articular cartilage and subchondral bone of rats were elevated in the aged group compared with the young group(P<0.05)and decreased in the pulsed electromagnetic field group compared with the old group(P<0.05).To conclude,pulsed electromagnetic fields may improve osteoarthritis in aged rats by inhibiting chondrocyte senescence,alleviating articular cartilage degradation and inhibiting subchondral bone osteoporosis through suppressing the expression of P53/P21.
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Colorectal cancer is a common clinical malignant tumor. As the main therapeutic method of colorectal cancer, radiotherapy has a good inhibitory effect on tumor progression. In recent years, because of its good physical and biological advantages, carbon ion has shown better clinical efficacy than traditional radiotherapy in the treatment of local recurrence or distant metastasis of colorectal cancer. In this article, basic and clinical studies related to the efficacy of carbon ion therapy for the recurrence of colorectal cancer in recent years were reviewed, aiming to provide theoretical basis for preventing and reducing adverse reactions after radiotherapy and prolonging the survival of colorectal cancer patients.
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Objective To explore the mechanism by which CD137 signal regulates the aging of vas-cular smooth muscle cells(VSMCs).Methods Thirty 8-week-old male C57BL/6J mice were ran-domly divided into a young group(8 weeks old)and an aged group(80 weeks old),with 30 mice in each group.After corresponding periods of feeding,the mice were euthanized,and the plasma and aortic blood vessels were isolated.In the cell experiments,normal VSMCs were divided into a control group,bleomycin(BLM)group,combined agonist group,and combined inhibitor group.The cellular senescence level of VSMCs was assessed using a cellular senescence β-galactosidase staining kit.Western blotting and PCR were employed to examine the expression of senescence-related proteins in tissues and cells,while ELISA was utilized to measure the expression of senes-cence-related inflammatory factors.Results The expression of CD137 and γ-H2AX in the aorta was significantly higher,while that of PCNA was obviously lower in the aged group than the young group(P<0.05).The plasma level of CD137 was notably higher in the aged group than the young group(154.0±4.1 pg/ml vs 98.0±2.3 pg/ml,P<0.05).Compared with the normal control group,there were significantly more aged VSMCs in the BLM group(P<0.05).While,treatment of combined agonist resulted in larger amount of aged VSMCs when compared with the BLM group(P<0.05),which was reversed by combined inhibitor treatment(P<0.05).The levels of TNF-α,IL-6 and IL-1β were significantly elevated in the BLM group than the normal control group(P<0.05).The combined agonist group had even higher levels of TNF-α,IL-6,and IL-1βthan the BLM group(P<0.05),but the levels were decreased in the combined inhibitor group(P<0.05).Compared with the normal control group,the expression of Bcl-2,γ-H2AX,P53,and P21 were significantly increased in the BLM group,combined agonist group,and combined inhibi-tor group,while that of PCNA was significantly decreased(P<0.05).Compared with the BLM group,the expression of P53 and P21 in the combined agonist group showed an increase(P<0.05),and the expression of P53 was significantly decreased in the combined inhibitor group(P<0.05).Conclusion CD137 signal regulates the P53/P21 pathway to promote VSMC aging.
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Objective:To establish a single-tube, one-step method for detecting and identifying 16 high-risk human papillomavirus (HR-HPV) subtypes (HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, 73, 82) and genotyping p53 (rs1042522) and RB1 (rs3092905) in cervical cells, using high-throughput two-dimensional PCR (2D-PCR) technology. Methods:Applied Research. Specific primers were designed according to the DNA sequences of the 16 different HR-HPV subtypes, p53, and RB1 genes, with the target genes p53 and RB1 serving as internal references to assess the success of sample collection and PCR amplification. In three fluorescent detection channels, upstream primers labeled with corresponding tags were used for different HR-HPV subtypes, p53, and RB1, constructing a comprehensive 2D-PCR detection system. Using this method, 804 cervical brush samples collected from the gynecology outpatient department of Changzhou First People′s Hospital from December 2022 to August 2023 were tested. The test results were compared for consistency with PCR-reverse dot blot assay, flow cytometric fluorescence hybridization assay, and single-plex real-time quantitative PCR assay, respectively. Meanwhile, the genotypes of p53 and RB1 were detected using Sanger sequencing. The Kappa test was applied to determine the consistency between 2D-PCR method and other methods. Results:2D-PCR accurately discriminated and identified the genotypes of 16 HR-HPV types and p53, RB1 through characteristic melting valleys in the FAM, HEX, and Alexa Fluor568 channels. 2D-PCR showed high consistency with PCR-reverse dot blot assay, with a Kappa value of 0.699, even higher consistency with flow cytometric fluorescence hybridization assay, with a Kappa value of 0.793, and the highest consistency with single-plex quantitative PCR, with a Kappa value of 0.880 (95% CI 0.862-0.907). Using Sanger sequencing as the gold standard, the accuracy of 2D-PCR method in detecting p53 and RB1 genotypes is 100%. The distribution frequencies of the three genotypes (G/G, G/C, and C/C) at the p53 rs1042522 locus were 32.09% (258/804), 49.88% (401/804) and 18.03% (145/804), respectively, while all detected genotypes at the RB1 rs3092905 locus were A/A. Conclusion:This study successfully developed a 2D-PCR method for the identification and genotyping of high-risk human papillomavirus types and related tumor suppressor genes p53 and RB1 for cervical cancer.
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【Objective】 To explore the expression of PCDH9 loss in regulating cell cycle and promoting tumor progression. 【Methods】 The clinical records of 127 cases of prostate cancer treated during 2018 and 2023 were collected, including 87 paraffin tissue samples from the G4-5 group and 40 from the G1-3 group. The expressions of PCDH9, p53, Rb and STAT3 were detected with immunohistochemical staining, and the relationship between their expressions and clinicopathological characteristics was analyzed. 【Results】 The expression deletion rate of PCDH9 in prostate cancer tissues in G4-5 group (44.8% vs.7.5%) was significantly higher than that in G1-3 group (P<0.001). The positive expression rates of p53 and STAT3 were 34.5% and 89.7%, respectively, and the expression loss rate of Rb was 27.6% in G4-5 group. The expression loss rates of PCDH9 and Rb were associated with neuroendocrine-like histological morphology, nerve invasion and vascular invasion (P<0.05). In G4-5 group of prostate cancer, PCDH9 expression was positively correlated with the expressions of p53 (r=0.345, P<0.05), Rb (r=0.503, P<0.05) and STAT3 (r=0.224, P<0.05). 【Conclusion】 PCDH9 is prone to loss of expression in high-group prostate cancer tissues, especially in cases with neuroendocrine-like histological morphology, which may regulate the cell cycle through the STAT3 signaling pathway, thereby promoting tumor progression.
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Lung cancer, the most common malignant tumor, is characterized by a complex pathogenesis and high malignancy, and poses a significant threat to the health and lives of affected individuals. p53 signaling pathway plays a crucial role in the progression of lung cancer and is considered one of the potential targets for targeted therapy. In recent years, multiple studies have indicated that traditional Chinese medicine (TCM) can exert anticancer effects by modulating the p53 signaling pathway. Based on this, this article systematically summarizes the current status and progress of research on TCM intervening in lung cancer by regulating p53 signaling pathway. It was found that TCM formula and preparations, such as Qingjin desheng tablet, Tiaoqi xiaoji decoction, Bufei tongluo jiedu formula, Jianpi bushen formula and Yiqi fuzheng jiedu formula, can promote autophagy and apoptosis of lung cancer cells, inhibit the growth and metastasis of lung cancer cells and strengthen the immune function of the body by activating p53 signaling pathway, thereby inhibiting lung cancer. TCM monomers, such as pseudoginsenoside-Rh2, saikosaponin D, polyphyllin Ⅶ, dendrobiine, sophoridine, gambogic acid, triptolide and triptolide succinate monoester YJ-4, can accelerate cell apoptosis, inhibit the proliferation of lung cancer cells and regulate cell cycle by activating p53 signaling pathway, thereby inhibiting lung cancer.
ABSTRACT
Lung cancer, the most common malignant tumor, is characterized by a complex pathogenesis and high malignancy, and poses a significant threat to the health and lives of affected individuals. p53 signaling pathway plays a crucial role in the progression of lung cancer and is considered one of the potential targets for targeted therapy. In recent years, multiple studies have indicated that traditional Chinese medicine (TCM) can exert anticancer effects by modulating the p53 signaling pathway. Based on this, this article systematically summarizes the current status and progress of research on TCM intervening in lung cancer by regulating p53 signaling pathway. It was found that TCM formula and preparations, such as Qingjin desheng tablet, Tiaoqi xiaoji decoction, Bufei tongluo jiedu formula, Jianpi bushen formula and Yiqi fuzheng jiedu formula, can promote autophagy and apoptosis of lung cancer cells, inhibit the growth and metastasis of lung cancer cells and strengthen the immune function of the body by activating p53 signaling pathway, thereby inhibiting lung cancer. TCM monomers, such as pseudoginsenoside-Rh2, saikosaponin D, polyphyllin Ⅶ, dendrobiine, sophoridine, gambogic acid, triptolide and triptolide succinate monoester YJ-4, can accelerate cell apoptosis, inhibit the proliferation of lung cancer cells and regulate cell cycle by activating p53 signaling pathway, thereby inhibiting lung cancer.
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ObjectiveTo observe the effects of Sargassum and Glycyrrhizae Radix et Rhizoma incompatible pair with the Haizao Yuhutang (HYT) on oxidative stress in the liver of goiter rats under the condition of 2 times the dose limit of the Pharmacopoeia of the People's Republic of China 2020. MethodA total of 128 male Wistar rats were randomly divided into a blank group, a model group, a euthyrox group (20 μg·kg-1), a HYT group (12.06 g·kg-1), a HYT without Sargassum (HYT-H) group (9.90 g·kg-1), a HYT without Glycyrrhizae Radix et Rhizoma (HYT-G) group (10.26 g·kg-1), a HYT without Sargassum and Glycyrrhizae Radix et Rhizoma (HYT-HG) group (8.10 g·kg-1), and a Sargassum and Glycyrrhizae Radix et Rhizoma (HG) group (3.96 g·kg-1). The blank group was given deionized water by gavage, and the others were given propylthiouracil (PTU) to replicate the goiter pathological model. Euthyrox was taken as a positive control drug, and the rest of the Chinese medicine groups were given the corresponding decoction by gavage, the material was collected 12 hours after the last dose. The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and the contents of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), reactive oxygen species (ROS) in liver tissue were detected in each group. The pathological changes in the liver were observed via hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was utilized to detect the mRNA expressions of Kelch-like Ech-associated protein 1 (Keap1), nuclear factor E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), p53 and Caspase-3 in liver tissues. Western blot was adopted to detect the protein expressions of Nrf2 and HO-1 in liver tissues in oxidative stress-related signaling pathways. ResultCompared with control group, the model group showed significantly increased serum ALT level and contents of MDA and ROS in liver tissues (P<0.05, P<0.01), significantly reduced activities of SOD and GSH-Px in the liver (P<0.01), significantly increased mRNA expression of Keap1 (P<0.01), and significantly decreased mRNA and protein expressions of Nrf2 and HO-1 (P<0.05, P<0.01). Compared with the model group, the HYT group manifested significantly reduced serum levels of AST, ALT, and ALP (P<0.05, P<0.01), significantly reduced contents of MDA and ROS in liver tissue (P<0.01), significantly increased the activities of SOD and GSH-Px (P<0.01), significantly decreased mRNA expressions of Keap1, p53, and Caspase-3 (P<0.01), and significantly increased mRNA and protein expressions of Nrf2 and HO-1 (P<0.05, P<0.01). ConclusionUnder the condition of 2 times the dose limit of the Pharmacopoeia of the People's Republic of China 2020, Sargassum and Glycyrrhizae Radix et Rhizoma incompatible pair with the HYT on oxidative stress in the liver of goiter rats had different effects. The HYT that contains Sargassum and Glycyrrhizae Radix et Rhizoma has a protective effect on the liver of goiter rats, and the effect is better than that of the HG group, the euthyrox group, and the incomplete groups. Its mechanism may be related to activating the Nrf2/HO-1 signaling pathway to alleviate liver oxidative stress and inhibiting the p53/Caspase-3 signaling pathway to reduce hepatocyte apoptosis.