RÉSUMÉ
Drug resistance remains a significant challenge in the treatment of pancreatic cancer. The development of drug-resistant cell lines is crucial to understanding the underlying mechanisms of resistance and developing novel drugs to improve clinical outcomes. Here, a novel pancreatic cancer cell line, PDAC-X1, derived from Chinese patients has been established. PDAC-X1 was characterized by the immune phenotype, biology, genetics, molecular characteristics, and tumorigenicity. In vitro analysis revealed that PDAC-X1 cells exhibited epithelial morphology and cell markers (CK7 and CK19), expressed cancer-associated markers (E-cadherin, Vimentin, Ki-67, CEA, CA19-9), and produced pancreatic cancer-like organs in suspension culture. In vivo analysis showed that PDAC-X1 cells maintained tumorigenicity with a 100% tumor formation rate. This cell line exhibited a complex karyotype, dominated by subtriploid karyotypes. In addition, PDAC-X1 cells exhibited intrinsic multidrug resistance to multiple drugs, including gemcitabine, paclitaxel, 5-fluorouracil, and oxaliplatin. In conclusion, the PDAC-X1 cell line has been established and characterized, representing a useful and valuable preclinical model to study the underlying mechanisms of drug resistance and develop novel drug therapeutics to improve patient outcomes.
Sujet(s)
Carcinome du canal pancréatique , Multirésistance aux médicaments , Résistance aux médicaments antinéoplasiques , Tumeurs du pancréas , Humains , Lignée cellulaire tumorale , Carcinome du canal pancréatique/anatomopathologie , Carcinome du canal pancréatique/traitement médicamenteux , Carcinome du canal pancréatique/génétique , Carcinome du canal pancréatique/métabolisme , Tumeurs du pancréas/anatomopathologie , Tumeurs du pancréas/traitement médicamenteux , Tumeurs du pancréas/génétique , Tumeurs du pancréas/métabolisme , Animaux , Souris , Multirésistance aux médicaments/génétique , Tests d'activité antitumorale sur modèle de xénogreffe , Mâle , Femelle , Antinéoplasiques/pharmacologie , Antinéoplasiques/usage thérapeutique , Marqueurs biologiques tumoraux/génétique , Marqueurs biologiques tumoraux/métabolisme , , Désoxycytidine/analogues et dérivés , Désoxycytidine/pharmacologie , Désoxycytidine/usage thérapeutiqueRÉSUMÉ
In this study, a novel pancreatic cancer cell line, termed pancreatic ductal adenocarcinoma (PDAC)-X3 cell line, was successfully derived from the primary tumor. Comprehensive analyses of its malignant phenotype, molecular properties, specific biomarkers, and histological features confirmed that PDAC-X3 cells serve as a valuable model for investigating the underlying mechanisms driving pancreatic carcinogenesis and advancing potential therapeutic strategies. The newly established cell line was continuously cultured for over 12 months and was stably passaged through more than 50 generations. Morphologically, PDAC-X3 cells displayed characteristics typical of epithelial tumors. The population doubling time for PDAC-X3 cells was determined to be 50 h. Karyotype analysis revealed that 75% of PDAC-X3 cells presented as hypotriploid, while 25% were sub-tetraploid, with representative karyotypes being 53 and XY der (1) inv (9) der (22). In suspension culture, PDAC-X3 cells efficiently formed organoids. Upon inoculation into BALB/C nude mice, these cells initiated the development of xenograft tumors, achieving a tumor formation rate of 33%. Morphologically, these xenografted tumors closely resembled the primary tumor. Drug sensitivity assays indicated that PDAC-X3 cells exhibited resistance to oxaliplatin but demonstrated sensitivity to 5-Fluorouracil (5-FU), gemcitabine, and paclitaxel. Immunohistochemical analysis revealed that CK7, CK19, E-cadherin, Vimentin, CA19-9 were positively expressed in PDAC-X3 cells. Meanwhile, the expression rate for Ki-67 was 30%, and that for CEA was not detected. Our findings underscore that PDAC-X3 represents a novel pancreatic cancer cell line, positioning it as a valuable model for basic research and the advancement of therapeutic strategies against pancreatic cancer.
Sujet(s)
Carcinome du canal pancréatique , Fluorouracil , Souris de lignée BALB C , Souris nude , Tumeurs du pancréas , Tumeurs du pancréas/anatomopathologie , Tumeurs du pancréas/génétique , Humains , Carcinome du canal pancréatique/anatomopathologie , Carcinome du canal pancréatique/génétique , Lignée cellulaire tumorale , Animaux , Fluorouracil/pharmacologie , Paclitaxel/pharmacologie , Résistance aux médicaments antinéoplasiques/génétique , Souris , , Désoxycytidine/analogues et dérivés , Désoxycytidine/pharmacologie , Hétérogreffes , Oxaliplatine/pharmacologie , Transplantation tumorale , Peuples d'Asie de l'EstRÉSUMÉ
Chemotherapy resistance poses clinical challenges in pancreatic cancer treatment. Developing cell lines resistant to chemotherapy is crucial for investigating drug resistance mechanisms and identifying alternative treatment pathways. The genetic and biological attributes of pancreatic cancer depend on its aetiology, racial demographics and anatomical origin, underscoring the need for models that comprehensively represent these characteristics. Here, we introduce PDAC-X2, a pancreatic cancer cell line derived from Chinese patients. We conducted a comprehensive analysis encompassing the immune phenotype, biology, genetics, molecular characteristics and tumorigenicity of the cell line. PDAC-X2 cells displayed epithelial morphology and expressed cell markers (CK7 and CK19) alongside other markers (E-cadherin, Vimentin, Ki-67, CEA and CA19-9). The population doubling time averaged around 69 h. In vivo, PDAC-X2 cells consistently maintained their tumorigenicity, achieving a 100% tumour formation rate. Characterised by a predominantly tetraploid karyotype, this cell line exhibited a complex genetic markup. Notably, PDAC-X2 cells demonstrated resistance to multiple drugs, including gemcitabine, paclitaxel, 5-fluorouracil and oxaliplatin. In conclusion, PDAC-X2 presents an invaluable preclinical model. Its utility lies in facilitating the study of drug resistance mechanisms and the exploration of alternative therapeutic approaches aimed at enhancing the prognosis of this tumour type.
Sujet(s)
Carcinome du canal pancréatique , Résistance aux médicaments antinéoplasiques , Tumeurs du pancréas , Animaux , Femelle , Humains , Souris , Antinéoplasiques/pharmacologie , Antinéoplasiques/usage thérapeutique , Asiatiques , Carcinome du canal pancréatique/anatomopathologie , Carcinome du canal pancréatique/traitement médicamenteux , Carcinome du canal pancréatique/génétique , Lignée cellulaire tumorale , Multirésistance aux médicaments/génétique , Peuples d'Asie de l'Est , , Tumeurs du pancréas/anatomopathologie , Tumeurs du pancréas/traitement médicamenteux , Tumeurs du pancréas/génétique , Tumeurs du pancréas/métabolisme , Tests d'activité antitumorale sur modèle de xénogreffeRÉSUMÉ
OBJECTIVE: The development and current state of hemorrhagic fever with renal syndrome (HFRS) over the past 40 years are analyzed in this study, along with explored and discovered the hotspots and frontiers in the field, which serve as the foundation for future investigation. METHODS: CiteSpace and VOSviewer analysis software were used to visually analyze the literature data on HFRS from 1980 to 2022, including the annual number of publications, countries and research institutions, authors, co-cited literature and keywords. RESULTS: The number of pertinent papers published in the field of HFRS displayed an overall upward trend from 1980 to 2022. The United States, China, Germany, Sweden, and France are the top 5 countries in terms of publishing volume, with high intermediate centrality mainly concentrated in Europe and the United States. The top 10 co-occurring keywords were hemorrhagic fever, renal syndrome, infection, virus, epidemic, nephropathia epidemical, disease, hantavirus, outbreak, and transmission. According to keyword cluster analysis, there were 4 main research fields. In the HFRS-related study, there were mainly 21 notable keywords and "Korean hemorrhagic fever" had the highest hemorrhagic value (28.87). CONCLUSION: The United States, China, Germany, Sweden and other countries attached great importance to the HFRS-related research. Moreover, the collaboration between authors and institutions in various collaborator clusters should be strengthened. In recent decades, investigations have focused on the study of viral infection and the clinical symptoms and pathophysiology of HFRS. Future research may concentrate on factors affecting host population distribution and density, such as vaccine development and meteorological factors pertaining to virus transmission.
Sujet(s)
Fièvre hémorragique avec syndrome rénal , Humains , Fièvre hémorragique avec syndrome rénal/épidémiologie , Europe , Chine/épidémiologie , Allemagne , FranceRÉSUMÉ
Background: Graves' disease (GD) is an autoimmune thyroid disorder. Our previous study has demonstrated a significant decrease in flavone levels among children with GD compared to the control group. Puerarin, a well-known flavonoid with anti-inflammatory and antioxidant properties. We wanted to investigate its potential impact on GD pathogenesis, aiming to determine whether increasing puerarin intake could prevent or delay the onset of GD. Methods: Adenovirus with TSHR-289 subunit was used to establish a GD mice model, and mice were intragastrically administered with puerarin or sterilized water daily. Thyroid function and inflammatory cytokine levels were quantified using ELISA, lymphocyte subsets were analyzed via flow cytometry, oxidative stress (OS) markers were measured with a microplate reader, and the expression of pertinent signaling pathway proteins were assessed by Western blot. Results: The results demonstrated that puerarin treatment significantly decreased thyroxin levels and alleviated thyroid pathological changes in GD mice. Furthermore, the immune imbalance of GD mice was improved, as evidenced by reduced inflammatory indexes, elevated antioxidant levels, and decreased malondialdehyde (MDA) levels compared to untreated GD mice. Puerarin-treated GD mice exhibited significantly lower expressions of heat shock protein (HSP): HSP70, HSP90, phosphorylated extracellular regulated kinases (p-ERK) and phosphorylated protein kinase B (p-AKT) than untreated GD mice. Moreover, low dosage puerarin (400 mg/kg) was associated with a better protective effect than high dosage (1,200 mg/kg). Conclusions: Puerarin may have the potential to mitigate GD by inhibiting inflammatory and OS, through downregulating the expression of HSP70 and HSP90 and suppressing the activation of the PI3K/AKT/ERK signaling pathway. Furthermore, a lower dose exhibited superior protective effects compared to a higher dose.
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Mixed-type ampullary cancer is a distinct subtype of ampullary cancer that manifests a merging of the biological characteristics of both intestinal and pancreaticobiliary subtypes. The absence of established cell lines specific to this subtype has resulted in a concomitant scarcity of research on its tumorigenic mechanisms and the development of novel therapeutic modalities. The present study achieved the successful establishment of a novel mixed-type ampullary cancer cell line, designated DPC-X4 through primary culture techniques. Subsequent analyses pertaining to phenotypic characteristics, molecular profiling, biomarker identification, and histological features validated the DPC-X4 cell line as a potent model for delineating the pathogenesis of mixed-type ampullary cancer and facilitating the development of new pharmacological agents. This newly established cell line was subjected to continuous cultivation for 1 year, with stable passaging for over 50 generations. Notably, the DPC-X4 cell line manifested typical morphological features associated with epithelial tumors. Furthermore, the population doubling time for the DPC-X4 cell line was determined at 70 h. Short tandem repeat (STR) analysis confirmed that the DPC-X4 cell line exhibited a high genetic concordance with the primary tumor from the patient. Karyotypic profiling indicated an abnormal sub-triploid karyotype, with representative karyotypes of 57, XXY inv (9), 14p + , 15p + , der (17), + mar. The DPC-X4 cell line demonstrated a high capacity for efficient organoid formation under suspension culture conditions. In addition, the subcutaneous inoculation of DPC-X4 cells into NXG mice led to the formation of xenografted tumors. The results of drug sensitivity testing indicated that DPC-X4 cells were sensitive to paclitaxel and resistant to oxaliplatin, 5-fluorouracil, and gemcitabine. Immunohistochemistry revealed positive expression of CK7, CK19, and CK20 in DPC-X4 cells, while CDX2 demonstrated negative expression. In addition, positive expression of E-cadherin and vimentin was identified in DPC-X4 cells, with a proliferation index indicated by Ki-67 at 70%. The findings of our study establish DPC-X4 as a novel mixed-type ampullary cancer cell line, which can serve as a potential experimental model for exploring the pathogenesis of ampullary cancer and the development of therapeutic drugs.
Sujet(s)
Ampoule hépatopancréatique , Tumeurs du cholédoque , Tumeurs , Humains , Animaux , Souris , Marqueurs biologiques tumoraux/métabolisme , Ampoule hépatopancréatique/composition chimique , Ampoule hépatopancréatique/métabolisme , Ampoule hépatopancréatique/anatomopathologie , Tumeurs du cholédoque/génétique , Tumeurs du cholédoque/métabolisme , Tumeurs du cholédoque/anatomopathologie , Tumeurs/anatomopathologie , Lignée cellulaire , Lignée cellulaire tumoraleRÉSUMÉ
BACKGROUND: The pulmonary surfactant that lines the air-liquid surface within alveoli is a protein-lipid mixture essential for gas exchange. Surfactant lipids and proteins are synthesized and stored in the lamellar body (LB) before being secreted from alveolar type II (AT2) cells. The molecular and cellular mechanisms that regulate these processes are incompletely understood. We previously identified an essential role of general control of amino acid synthesis 5 like 1 (GCN5L1) and the biogenesis of lysosome-related organelle complex 1 subunit 1 (BLOS1) in surfactant system development in zebrafish. Here, we explored the role of GCN5L1 in pulmonary surfactant regulation. METHOD: GCN5L1 knockout cell lines were generated with the CRISPR/Cas9 system. Cell viability was analyzed by MTT assay. Released surfactant proteins were measured by ELISA. Released surfactant lipids were measured based on coupled enzymatic reactions. Gene overexpression was mediated through lentivirus. The RNA levels were detected through RNA-sequencing (RNA-seq) and quantitative reverse transcription (qRT)- polymerase chain reaction (PCR). The protein levels were detected through western blotting. The cellular localization was analyzed by immunofluorescence. Morphology of the lamellar body was analyzed through transmission electron microscopy (TEM), Lysotracker staining, and BODIPY phosphatidylcholine labeling. RESULTS: Knocking out GCN5L1 in MLE-12 significantly decreased the release of surfactant proteins and lipids. We detected the downregulation of some surfactant-related genes and misregulation of the ROS-Erk-Foxo1-Cebpα axis in mutant cells. Modulating the activity of the axis or reconstructing the mitochondrial expression of GCN5L1 could partially restore the expression of these surfactant-related genes. We further showed that MLE-12 cells contained many LB-like organelles that were lipid enriched and positive for multiple LB markers. These organelles were smaller in size and accumulated in the absence of GCN5L1, indicating both biogenesis and trafficking defects. Accumulated endogenous surfactant protein (SP)-B or exogenously expressed SP-B/SP-C in adenosine triphosphate-binding cassette transporterA3 (ABCA3)-positive organelles was detected in mutant cells. GCN5L1 localized to the mitochondria and LBs. Reconstruction of mitochondrial GCN5L1 expression rescued the organelle morphology but failed to restore the trafficking defect and surfactant release, indicating specific roles associated with different subcellular localizations. CONCLUSIONS: In summary, our study identified GCN5L1 as a new regulator of pulmonary surfactant that plays a role in the biogenesis and positioning/trafficking of surfactant-containing LBs.
Sujet(s)
Surfactants pulmonaires , Animaux , Souris , Pneumocytes/métabolisme , Corps lamellaires , Lipides , Surfactants pulmonaires/métabolisme , ARN , Tensioactifs , Danio zébré/métabolismeRÉSUMÉ
Malignant melanoma is the most aggressive form of skin cancer, and it is characterized by poor prognosis in patients with metastatic diseases. Accurate prediction of prognosis is crucial for therapeutic decisions. In this study, bioinformatics analysis was used to explore the prognostic value of growth factor receptor-bound protein 2-associated binding protein 3 (GAB3) mRNA. RNA transcriptome sequencing data and clinical data from The Cancer Genome Atlas and genotype-tissue expression (GTEx) were analyzed for differentially expressed genes in high and low GAB3 mRNA expression groups in melanoma. Performing gene enrichment analysis and constructing protein-protein interaction networks. High expression of GAB3 was significantly correlated with a lower T stage, melanoma Clark level, Breslow depth, and melanoma ulceration. And high GAB3 expression was also associated with better progression-free interval in T1 and T2 stages and N0 stage and longer overall survival in T1 and T2 stages, N0 stage, and N1 stage. GAB3 promoted high levels of infiltration of macrophages and activated natural killer cells in melanoma. High expression of GAB3 predicted a positive prognosis in early-stage melanoma that may be mediated by the anticancer immune response.
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Mélanome , Tumeurs cutanées , Humains , Mélanome/anatomopathologie , Tumeurs cutanées/anatomopathologie , ARN messager/génétique , Pronostic , Transcriptome , Protéine adaptatrice GRB2/génétique , Protéine adaptatrice GRB2/métabolisme , Protéines adaptatrices de la transduction du signal/génétique , Protéines adaptatrices de la transduction du signal/métabolismeRÉSUMÉ
Objective: To summarise the clinical characteristics of patients with Stevens-Johnson syndrome/toxic epidermal necrolysis syndrome (SJS/TEN) and analyse the efficacy and safety of systemic glucocorticoid therapy. Methods: This study was a retrospective study of 56 patients with SJS/TEN who had been systematically treated with glucocorticoids in the dermatology ward of Peking University Third Hospital from 2010 to 2020. The clinical characteristics, treatment regimen, effects on underlying diseases, incidence and outcome of hormone-related adverse reactions and skin lesion prognosis were summarised and analysed for each patient. Results: â The allergenic drugs were found to be antibiotics (31.51%), antipyretic and analgesics (21.92%), traditional Chinese medicines and health products (15.07%) and neuropsychiatric drugs (13.70%). â¡ Based on the 56 patients' scores of toxic epidermal necrosis at admission, the actual mortality rate was 1.8% (1/56), which was significantly lower than the average expected mortality rate of 15.0% (P = 0.032; standardised mortality ratio = 0.13; 95% confidence interval: 0.00-0.53). ⢠A total of 33 patients (58.9%) had underlying diseases, of which 10 patients (30.3%) had underlying diseases that fluctuated during treatment but stabilised after symptomatic treatment. ⣠During treatment, 73.2% (41/56) of patients had complications that may have been related to systemic glucocorticoids; 97.6% (40/41) had mild symptoms, and 92.7% (38/41) had improved/recovered complications at the time of discharge. Conclusion: â Antibiotics are still the most common sensitising drugs, and traditional Chinese medicine and health products are also common sensitising drugs. â¡ Early systemic application of medium- to high-dose glucocorticoids is effective in the treatment of SJS/TEN, and it is beneficial in reducing mortality. ⢠The short-term application of medium- to high-dose hormone therapy for SJS/TEN has little effect on underlying diseases. The related complications are mostly mild, and the treatment is safe.
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OBJECTIVE: To investigate current situation of facial wrinkles of male construction workers in Beijing area and to discuss the correlative factors. MATERIALS AND METHODS: A total of 149 male construction works and 63 male non-construction workers in Beijing were required to complete a questionnaire on their exposure to sunlight, dust, noise, and heat in their workplace environment. Their facial wrinkle scores were measured by VISIA Complexion Analysis System. The two-sample t test, chi-square test, and multiple linear regression were used for statistical analysis RESULTS: The exposure to sunlight, dust, noise, and heat of construction workers was significantly higher than that of non-construction workers (P < .01). The wrinkle score of construction workers between 20 and 29 years old was significantly higher than that of non-construction workers (t = 4.077, P < .01). The facial wrinkle score of construction workers(r = 0.657, P < .01) and non-construction workers (r = 0.681, P < .01) was both positively correlated with age. The wrinkle score of construction workers was related to age, sunlight, and noise(P < .01). CONCLUSION: The wrinkle score of male construction workers between 20 and 29 years old is significantly higher than that of non-construction workers in Beijing. Age, sunlight, and noise were the main influencing factors of wrinkle.
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Industrie de la construction , Vieillissement de la peau , Adulte , Face , Humains , Mâle , Lumière du soleil , Enquêtes et questionnaires , Jeune adulteRÉSUMÉ
Mammalian mitochondrial tRNA (mt-tRNA) plays a central role in the synthesis of the 13 subunits of the oxidative phosphorylation complex system (OXPHOS). However, many aspects of the context-dependent expression of mt-tRNAs in mammals remain unknown. To investigate the tissue-specific effects of mt-tRNAs, we performed a comprehensive analysis of mitochondrial tRNA expression across five mice tissues (brain, heart, liver, skeletal muscle, and kidney) using Northern blot analysis. Striking differences in the tissue-specific expression of 22 mt-tRNAs were observed, in some cases differing by as much as tenfold from lowest to highest expression levels among these five tissues. Overall, the heart exhibited the highest levels of mt-tRNAs, while the liver displayed markedly lower levels. Variations in the levels of mt-tRNAs showed significant correlations with total mitochondrial DNA (mtDNA) contents in these tissues. However, there were no significant differences observed in the 2-thiouridylation levels of tRNALys, tRNAGlu, and tRNAGln among these tissues. A wide range of aminoacylation levels for 15 mt-tRNAs occurred among these five tissues, with skeletal muscle and kidneys most notably displaying the highest and lowest tRNA aminoacylation levels, respectively. Among these tissues, there was a negative correlation between variations in mt-tRNA aminoacylation levels and corresponding variations in mitochondrial tRNA synthetases (mt-aaRS) expression levels. Furthermore, the variable levels of OXPHOS subunits, as encoded by mtDNA or nuclear genes, may reflect differences in relative functional emphasis for mitochondria in each tissue. Our findings provide new insight into the mechanism of mt-tRNA tissue-specific effects on oxidative phosphorylation.
Sujet(s)
Mitochondries , ARN de transfert , Animaux , Noyau de la cellule/métabolisme , Souris , Spécificité d'organe , Phosphorylation oxydative , Maturation post-transcriptionnelle des ARNRÉSUMÉ
BACKGROUND AND AIM: Transient global amnesia (TGA) was first described by Bender in 1956 and is characterized by sudden, temporary, and anterograde memory loss. This study aimed to explore the possible mechanisms of and lesions responsible for TGA. METHODS: Retrospective data were collected from all patients with TGA admitted to Zhongshan Hospital, affiliated with Xiamen University, between October 1, 2011, and October 30, 2018. Information about the TGA condition, previous history, and clinical examination of the TGA and control groups was recorded. Functional magnetic resonance imaging was performed on the patients to explore the possible lesions responsible for TGA. RESULTS: A total of 73 patients with TGA and 73 age- and gender-matched controls were included in the analysis. The differences in the migraine history (9/2, p = 0.038) were statistically significant in both groups, but no statistically significant difference was observed regarding the history of hypertension, diabetes, and other diseases. In addition, seven patients with TGA had lesions located in the hippocampal CA1 region; the dome column and hippocampal CA1 region exist in the same functional loop and play a synergistic role. The average follow-up period in the groups was 36 months. During the follow-up period, no significant differences in cerebral infarction, cerebral hemorrhage, CHD, or TGA attack between the groups were observed. CONCLUSION: Migraine may be a risk factor of TGA, and cerebral infarction may be one of the pathogeneses. The brain area responsible for TGA may involve a memory loop comprising the hippocampal CA1 region and the fornix column among other parts.
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Defects in the posttranscriptional modifications of mitochondrial tRNAs have been linked to human diseases, but their pathophysiology remains elusive. In this report, we investigated the molecular mechanism underlying a deafness-associated tRNAIle 4295A>G mutation affecting a highly conserved adenosine at position 37, 3' adjacent to the tRNA's anticodon. Primer extension and methylation activity assays revealed that the m.4295A>G mutation introduced a tRNA methyltransferase 5 (TRMT5)-catalyzed m1G37 modification of tRNAIle. Molecular dynamics simulations suggested that the m.4295A>G mutation affected tRNAIle structure and function, supported by increased melting temperature, conformational changes and instability of mutated tRNA. An in vitro processing experiment revealed that the m.4295A>G mutation reduced the 5' end processing efficiency of tRNAIle precursors, catalyzed by RNase P. We demonstrated that cybrid cell lines carrying the m.4295A>G mutation exhibited significant alterations in aminoacylation and steady-state levels of tRNAIle. The aberrant tRNA metabolism resulted in the impairment of mitochondrial translation, respiratory deficiency, decreasing membrane potentials and ATP production, increasing production of reactive oxygen species and promoting autophagy. These demonstrated the pleiotropic effects of m.4295A>G mutation on tRNAIle and mitochondrial functions. Our findings highlighted the essential role of deficient posttranscriptional modifications in the structure and function of tRNA and their pathogenic consequence of deafness.
Sujet(s)
Surdité neurosensorielle/génétique , Mutation ponctuelle , ARN de transfert de l'isoleucine/génétique , Adénosine triphosphate/biosynthèse , Adulte , Protéines d'archée/métabolisme , Autophagie , Séquence nucléotidique , Lignée cellulaire , ADN mitochondrial/génétique , Ethnies/génétique , Femelle , Pléiotropie , Surdité neurosensorielle/ethnologie , Humains , Isoleucine/métabolisme , Mâle , Hérédité maternelle , Potentiel de membrane mitochondriale , Methanocaldococcus/enzymologie , Méthylation , Adulte d'âge moyen , Mitochondries/métabolisme , Simulation de dynamique moléculaire , Conformation d'acide nucléique , Phosphorylation oxydative , Pedigree , Biosynthèse des protéines , Maturation post-transcriptionnelle des ARN , Protéines recombinantes/métabolisme , Aminoacylation des ARN de transfert , Jeune adulte , T-RNA methyltransferases/métabolismeRÉSUMÉ
The tissue specificity of mitochondrial tRNA mutations remains largely elusive. In this study, we demonstrated the deleterious effects of tRNAThr 15927G>A mutation that contributed to pathogenesis of coronary artery disease. The m.15927G>A mutation abolished the highly conserved base-pairing (28C-42G) of anticodon stem of tRNAThr. Using molecular dynamics simulations, we showed that the m.15927G>A mutation caused unstable tRNAThr structure, supported by decreased melting temperature and slower electrophoretic mobility of mutated tRNA. Using cybrids constructed by transferring mitochondria from a Chinese family carrying the m.15927G>A mutation and a control into mitochondrial DNA (mtDNA)-less human umbilical vein endothelial cells, we demonstrated that the m.15927G>A mutation caused significantly decreased efficiency in aminoacylation and steady-state levels of tRNAThr. The aberrant tRNAThr metabolism yielded variable decreases in mtDNA-encoded polypeptides, respiratory deficiency, diminished membrane potential and increased the production of reactive oxygen species. The m.15927G>A mutation promoted the apoptosis, evidenced by elevated release of cytochrome c into cytosol and increased levels of apoptosis-activated proteins: caspases 3, 7, 9 and PARP. Moreover, the lower wound healing cells and perturbed tube formation were observed in mutant cybrids, indicating altered angiogenesis. Our findings provide new insights into the pathophysiology of coronary artery disease, which is manifested by tRNAThr mutation-induced alterations.
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Mitochondries/génétique , Néovascularisation pathologique/génétique , ARN de transfert de la thréonine/composition chimique , ARN de transfert/génétique , Apoptose/génétique , Maladie des artères coronaires/génétique , Maladie des artères coronaires/anatomopathologie , ADN mitochondrial/génétique , Cellules endothéliales de la veine ombilicale humaine , Humains , Mitochondries/anatomopathologie , Mutation , Conformation d'acide nucléique , ARN de transfert de la thréonine/génétique , Espèces réactives de l'oxygène/métabolismeRÉSUMÉ
Graphene is a hot star in materials science with various potential application aspects, including in Li-ion battery and supercapacitor. The burst of scientific papers in this area seems to validate the performance of graphene, but also arouses large dispute. Herein, we share our judgment of these trends to all, encouraging the discussion and enhancing the understanding of the structure-performance relationship of graphene.
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OBJECTIVE: To investigate the effect of oridonin on the human acute lymphocytic leukemia cell line Jurkat and its mechanism. METHODS: Jurkat cells were cultured in vitro and treated with various concentrations (0, 1.25, 2.5, 5, and 10â µmol/L) of oridonin for different lengths of time (24, 48, and 72â hours). The proliferation of Jurkat cells was analyzed by MTT assay. The changes in nuclear morphology were evaluated by fluorescence microscopy at 12 hours after treatment with various concentrations of oridonin. The expression levels of Brg1, P53, and C-myc were determined by semi-quantitative Western blot in Jurkat cells treated with various concentrations of oridonin for 24â hours or 5â µmol/L oridonin for various lengths of time (0, 2, 6, 12, and 24â hours). The expression levels of P53 and C-myc and proliferation of Jurkat cells were evaluated after Brg1 expression was knocked down by Brg1-specific siRNA. RESULTS: Compared with the control group, the proliferation of oridonin-treated Jurkat cells was significantly inhibited in a concentration- and time-dependent manner (P<0.05). According to the florescence microscopic analysis, oridonin treatment led to nuclear pyknosis in Jurkat cells. Compared with the control group, Jurkat cells treated with 5â µmol/L oridonin had reduced expression of Brg1 and C-myc but elevated expression of P53. Brg1 knock-down led to a significant reduction in proliferation of Jurkat cells (P<0.05), up-regulated expression of P53, and down-regulated expression of C-myc. CONCLUSIONS: Oridonin can inhibit the proliferation of Jurkat cells, probably via the Brg1 signaling pathway.
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Antinéoplasiques d'origine végétale/pharmacologie , Helicase/physiologie , Diterpènes de type kaurane/pharmacologie , Protéines nucléaires/physiologie , Facteurs de transcription/physiologie , Prolifération cellulaire/effets des médicaments et des substances chimiques , Helicase/analyse , Relation dose-effet des médicaments , Régulation négative , Humains , Cellules Jurkat , Protéines nucléaires/analyse , Leucémie-lymphome lymphoblastique à précurseurs B et T/traitement médicamenteux , Protéines proto-oncogènes c-myc/analyse , Transduction du signal/physiologie , Facteurs de transcription/analyse , Protéine p53 suppresseur de tumeur/analyseRÉSUMÉ
Objective To comparatively observe clinical efficacies of Fusidic Acid Cream (FAC) , Longzhu Ointment (LO) , and their combination of minocycline hydrochloride for treating facial acne vulgaris. Methods Totally 186 patients with acne vulgaris were randomly assigned to the FAC group (103 cases) and the LO group (83 cases). Each group was further divided into two subgroups ac- cording to the severity of acne: single treatment group and united treatment group. Patients with mild ac- ne vulgaris in the FAC group received FAC alone (39 cases) , and those with severe acne vulgaris in the FAC group received FAC and minocycline hydrochloride (64 cases). Patients with mild acne vulgaris in the LO group received LO alone (27 cases) , and those with severe acne vulgaris in the LO group received LO and minocycline hydrochloride. The therapeutic course for all was 4 weeks, with one return vis- it once per week. Grading of skin lesions was assessed by global acne grading system (GAGS). Clinical improvement was evaluated. Skin spots, red areas, and other data were statistically analyzed by VISIA skin analyzer. Results GAGS score was statistically different between before and after treatment in the FAC group and the LO group (P <0. 05). The total effective rate was 64. 1% (25)39) in single treatment group of the FAC group and 66. 7% (18/27) in single treatment group of the LO group, but with no statisti- cal difference between the two groups (Χ² =0. 09, P >0. 05). The total effective rate was 70. 3% (45/64) in united treatment group of the FAC group and 62. 5% (35/56) in united treatment group of the LO group, but with no statistical difference between the two groups (Χ² =0. 04, P >0. 05). Results of VISIA showed, compared with before treatment, statistical difference existed in red area of single treatment group of the FAC group and the LO group (P <0. 05). Statistical difference existed in ultraviolet rays, red area, sclererythrin of united treatment group of the FAC group and the LO group (P <0. 05). Conclusions FAC and LO could effectively control the inflammation of acne. LO had a rapid onset. Combined with minocy- cline hydrochloride, FAC could significantly reduce the secretion of fats, and LO could defense against ultraviolet more significantly.
Sujet(s)
Acné juvénile , Antibactériens , Médecine traditionnelle d'Asie orientale , Acné juvénile/traitement médicamenteux , Antibactériens/usage thérapeutique , Humains , Minocycline/usage thérapeutique , Résultat thérapeutiqueRÉSUMÉ
BACKGROUND: Monilethrix is an autosomal dominant hair disorder characterized clinically by alopecia and follicular papules. In this study, we collected a Han monilethrix family to detect the mutations in patients and investigated the correlation between the genotype and phenotype of monilethrix. METHODS: In this study, we identified a Chinese family with monilethrix through light microscopic and scanning electron microscopic (SEM) examination. Genomic DNA from peripheral blood samples was prepared. DNA samples from controls and monilethrix patients were subject to polymerase chain reaction (PCR) amplification. Two pairs of primers were used to amplify the seventh exon of KRT86. Mutation screening of the PCR products was detected using direct sequencing. RESULTS: Light microscopic examination showed a regular alternate enlargement and narrow area. SEM examination showed that part of the cuticle of the nodules shed and disappeared gradually in the narrow area with granular protrusions on the surface similar to the erosion-like structure. Parallel longitudinal ridge and groovepattern appeared, and the ridges varied in width, like dead wood. A heterozygous transversion mutation c.1204G > A (p.E402K) in the seventh exon of KRT86 was identified in both patients. CONCLUSIONS: The mutation of extron 7 of KRT86 identified plays a major role in the pathogenesis of this pedigree with monilethrix, and is a mutation hot spot of KRT86. Further research is needed to explore the relationship between the phenotype and the mutation of the type II hair keratin gene KRT86 of monilethrix.
Sujet(s)
Kératines spécifiques du cheveu/génétique , Kératines de type II/génétique , Monilethrix/génétique , Mutation , Asiatiques/génétique , Chine/ethnologie , Humains , Microscopie électrochimique à balayage , Monilethrix/étiologie , Monilethrix/anatomopathologieRÉSUMÉ
OBJECTIVE: To establish a HPLC-MS/MS method for the determination of vitexin in rat plasma and its pharmacokinetics. METHODS: The HPLC-MS/MS method used Capcell Pak C18 column (50 mm x 2.0 mm I. D., 5 microm). The mobile phase was methanol and water (95:5, V/V, containing 0.1% formic acid) at a flow rate of 0.2 mL/min. Electrospray ionization (ESI) in negative ion mode and multiple reaction monitoring (MRM) was used for the quantification of vitexin with a monitored transitions m/z 431-->311 for vitexin and m/z 269-->225 for internal standard (I. S., emodin). RESULTS: Linear calibration curves were obtained over the concentration range of 0.5-2000 ng/mL (r = 0.9960) with the lowest limit of quantification (LLOQ) of 0.5 ng/mL. The recovery was in the range of 76.1%-89.0%. The relative standard deviations for the intra-day and inter-day validation were less than 11%. CONCLUSION: The method is simple, accurate, fast, sensitive and suitable for the pharmacokinetic study of vitexin in rats.