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Objective: To investigate the characteristics of gene mutations in angioimmunoblastic T-cell lymphoma (AITL). Methods: Seventy-five AITL cases diagnosed at the Department of Pathology, Ruijin Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai, China from June 2021 to June 2023 were included. Their formalin-fixed and paraffin-embedded or fresh tissues were subject to targeted next generation sequencing (NGS). The sequencing data was collected, and the distribution and type of gene mutations were analyzed. Results: 492 potential driver mutations were identified in 74 out of the 84 genes. Targeted sequencing data for the 75 AITL patients showed that the genes with mutation frequencies of ≥10% were TET2 (89.3%), RHOA (57.3%), IDH2 (37.3%), DNMT3A (36.0%), KMT2C (21.3%), PLCG1 (12.0%), and KDM6B (10.7%). There were significant co-occurrence relationships between TET2 and RHOA, TET2 and IDH2, and RHOA and IDH2 gene mutations (P<0.05), respectively, while TET2 and KDM6B gene mutations were mutually exclusive (P<0.05). Conclusions: The study reveals the mutational characteristics of AITL patients using NGS technology, which would provide insights for molecular diagnosis and targeted therapy of AITL.
Sujet(s)
Lymphadénopathie angio-immunoblastique , Lymphome T , Humains , Lymphome T/génétique , Lymphome T/anatomopathologie , Chine , Lymphadénopathie angio-immunoblastique/diagnostic , Mutation , Taux de mutation , Jumonji Domain-Containing Histone Demethylases/génétiqueRÉSUMÉ
Objective: To investigate the clinicopathological characteristics and prognosis of high-grade B-cell lymphoma (HGBL) involving combined rearrangements of MYC, bcl-2 and bcl-6. Methods: A total of 1 138 cases of large B cell lymphoma (LBL) that were treated at the Ruijin Hospital Affiliated to Shanghai Jiaotong University School of Medicine from January 2017 to September 2020 were analyzed using fluorescence in situ hybridization (FISH) with probes against MYC, bcl-2 and bcl-6. The clinical and pathological data of the 45 patients with HGBL that had rearrangements of MYC and bcl-2 and/or bcl-6 were collected and retrospectively analyzed. Results: Among the 1 138 LBL, 45 (4.0%) cases had combined rearrangements of MYC, bcl-2 and/or bcl-6 that included 6 HGBL cases with MYC, bcl-2 and bcl-6 rearrangements, 14 HGBL cases with MYC and bcl-2 rearrangements, and 25 HGBL cases with MYC and bcl-6 rearrangements. Of these 45 patients, 29 patients were male, and 16 patients were female, aged 29 to 83 years. HGBL with MYC, bcl-2 and bcl-6 rearrangements and HGBL with MYC and bcl-2 rearrangement were reclassified as the germinal center B-cell (GCB) subtype using the Hans algorithm. HGBL with MYC and bcl-6 rearrangement were reclassified as the GCB subtype (68.0%) and the non-GCB subtype (32.0%). The vast majority of HGBL cases had a high Ki-67 proliferation index. Most HGBL patients had advanced stage disease with a high IPI score and an increased LDH level. Also, some patients had clinical features including elevated plasma ß2-microglobulin levels, B symptoms, and bone marrow involvement. The IPI scores and LDH levels were significantly different between the HGBL cases with MYC, bcl-2 and bcl-6 rearrangements and the HGBL cases with MYC and bcl-6 rearrangements (P<0.05). Compared with the HGBL cases with MYC, bcl-2 and bcl-6 rearrangements, the HGBL cases with MYC and bcl-2 or bcl-6 rearrangements had a lower incidence of bone marrow involvement (P<0.05). There were no significant differences in the prognosis among HGBL cases with MYC, bcl-2 and bcl-6 rearrangements, the cases with MYC and bcl-2 rearrangements, and the cases with MYC and bcl-6 rearrangements (P>0.05). Conclusions: HGBL with MYC, bcl-2 and/or bcl-6 rearrangements are rare types of B-cell lymphoma with high degree of malignancy and have a short overall survival. To reduce misdiagnosis and improve diagnostic accuracy, it is necessary to assess the patients' clinical features and conduct histopathological, immunohistochemical and FISH analyses.
Sujet(s)
Lymphome B diffus à grandes cellules , Protéines proto-oncogènes c-bcl-2 , Protéines proto-oncogènes c-bcl-6 , Protéines proto-oncogènes c-myc , Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Chine , Femelle , Réarrangement des gènes , Humains , Hybridation fluorescente in situ , Lymphome B diffus à grandes cellules/traitement médicamenteux , Lymphome B diffus à grandes cellules/génétique , Mâle , Adulte d'âge moyen , Pronostic , Protéines proto-oncogènes c-bcl-2/génétique , Protéines proto-oncogènes c-bcl-6/génétique , Protéines proto-oncogènes c-myc/génétique , Études rétrospectivesRÉSUMÉ
Objective: To investigate the relationship between the protein expression of C-MYC, bcl-2 and bcl-6 and the clinicopathological characteristics in patients with de novo CD5-positive diffuse large B cell lymphoma (CD5(+)DLBCL). Methods: Fifty seven cases of de novo CD5(+)DLBCL were collected at Ruijin Hospital Affiliated to Shanghai Jiaotong University School of Medicine from February 2013 to September 2018. The hematoxylin-eosin stained slides were reviewed, and immunohistochemical (IHC) staining and FISH were used to analyze the relationship between C-MYC, bcl-2, bcl-6 expression and the clinicopathologic characteristics of patients. Results: Among these 57 cases, 27 were male and 30 were female. The age of onset was 35-99 years old. The IHC expression rates of C-MYC, bcl-2 and bcl-6 were 50.9% (29/57), 84.2% (48/57), and 75.4% (43/57) respectively; and co-expression rate of C-MYC and bcl-2 proteins was 40.4 (23/57). There was no significant correlation between protein expression and patients' genders, clinical stage, the level of serum LDH,ß2 microglobulin, IPI,B symptoms, bone marrow involvement and central nervous system recurrence (P>0.05). Univariate analysis showed that the median OS of C-MYC negative patients was significantly longer than C-MYC positive patients (P<0.05); and the median OS of patients without double expression was significantly longer than that of patients with positive expression (P<0.05), and bcl-6 positive patients had longer median OS than bcl-6 negative patients (P<0.05). There was no significant correlation between prognosis and bcl-2 protein expression (P>0.05) . Cox multivariate analysis showed C-MYC protein expression was an independent predictor of OS in de novo CD5(+)DLBCL (P<0.05). Conclusions: Bcl-2 protein expression has no effect on the prognosis in de novo CD5(+)DLBCL whereas bcl-6 expression is correlated with good prognosis. C-MYC protein expression could be used as an independent and effective index to predict the prognosis of patients with de novo CD5(+)DLBCL.However, the relationship between protein expression and gene rearrangement of C-MYC, bcl-2 and bcl-6 needs to be further explored.
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Lymphome B diffus à grandes cellules , Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Chine , Femelle , Gènes myc , Humains , Mâle , Adulte d'âge moyen , Pronostic , Proto-oncogène Mas , Protéines proto-oncogènes c-bcl-2 , Protéines proto-oncogènes c-mycRÉSUMÉ
Objective: To observe the effects of 2-aminopurine (2-AP), a double-stranded RNA-dependent protein kinase (PKR) inhibitor, on organ function, plasma inflammatory factor expression and 7 days mortality in sepsis mice induced by cecal ligation puncture (CLP). Methods: Forty specific specific pathogen free C57BL/6 mice were randomly divided into sham group (n=10), CLP group (n=10), CLP+2-AP group (n=10) and 2-AP group (n=10). CLP was used to establish sepsis mice models.Peripheral blood serum was collected 24 hours after operation, alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum creatinine (Cr), blood urea nitrogen (BUN) and inflammatory factors (IL-1ß, IL-10 and TNF-α) were detected; peripheral blood and peritoneal lavage fluid were taken for bacterial clearance detection. Another 60 C57BL/6 mice were selected to observe the 7-day survival rate according to the above groups (n=15). Independent sample t test was used to compare the measurement data between groups. Results: The levels of ALT, AST, Cr and BUN in CLP Group and CLP+2-AP group were significantly higher than those in sham group (all P<0.001). The levels of ALT and AST in CLP+2-AP group were significantly lower than those in CLP Group (t=27.88, 11.33, both P<0.001); the levels of Cr and BUN in CLP+2-AP group were significantly lower than those in CLP Group (t=11.02, 7.15, bothP<0.001). Compared with sham group, the levels of pro-inflammatory (IL-1ß and TNF-α) and anti-inflammatory (IL-10) cytokines in CLP group were significantly higher (all P<0.001); the levels of IL-1ß and IL-10 in CLP+2-AP group were significantly lower (all P<0.001), but the levels of TNF-α in CLP+2-AP group were not significantly lower (P=0.33). The 7-day survival rate was 100% in sham group, 13.3% in CLP+2-AP group, 86.7% in 2-AP group and 20.0% in CLP+2-AP group. Inhibition of PKR activation slightly improved the trend of 7-days survival rate of CLP model mice (analysis by mantel Cox test, χ(2)=0.0012, P=0.97). Conclusion: In sepsis mice model, inhibition of PKR activity can reduce the expression of inflammatory factors in plasma, decrease bacterial load in blood and abdominal cavity, and protect organ function, which could suggest that inhibition of PKR activity has potential application in sepsis treatment.
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Sepsie , Animaux , Aspartate aminotransferases , Modèles animaux de maladie humaine , Souris , Souris de lignée C57BL , Facteur de nécrose tumorale alpha , eIF-2 KinaseRÉSUMÉ
Objective: To observe the effect of sepsis single-disease manage system on the improvement of sepsis treatment and the value in predicting mortality in patients with sepsis. Methods: A retrospective study was conducted. Patients with sepsis admitted to the Department of Surgical Intensive Care Unit of Sun Yat-Sen University First Affiliated Hospital from September 22, 2013 to May 5, 2015 were enrolled in this study. Sepsis single-disease manage system (Rui Xin clinical data manage system, China data, China) was used to monitor 25 clinical quality parameters, consisting of timeliness, normalization and outcome parameters. Based on whether these quality parameters could be completed or not, the clinical practice was evaluated by the system. The unachieved quality parameter was defined as suspicious parameters, and these suspicious parameters were used to predict mortality of patients with receiver operating characteristic curve (ROC). Results: A total of 1 220 patients with sepsis were enrolled, included 805 males and 415 females. The mean age was (59±17) years, and acute physiology and chronic health evaluation (APACHE â ¡) scores was 19±8. The area under ROC curve of total suspicious numbers for predicting 28-day mortality was 0.70; when the suspicious parameters number was more than 6, the sensitivity was 68.0% and the specificity was 61.0% for predicting 28-day mortality. In addition, the area under ROC curve of outcome suspicious number for predicting 28-day mortality was 0.89; when the suspicious outcome parameters numbers was more than 1, the sensitivity was 88.0% and the specificity was 78.0% for predicting 28-day mortality. Moreover, the area under ROC curve of total suspicious number for predicting 90-day mortality was 0.73; when the total suspicious parameters number was more than 7, the sensitivity was 60.0% and the specificity was 74.0% for predicting 90-day mortality. Finally, the area under ROC curve of outcome suspicious numbers for predicting 90-day mortality was 0.92; when suspicious outcome parameters numbers was more than 1, the sensitivity was 88.0% and the specificity was 81.0% for predicting 90-day mortality. Conclusion: The single center study suggests that this sepsis single-disease manage system could be used to monitor the completion of clinical practice for intensivist in managing sepsis, and the number of quality parameters failed to complete could be used to predict the mortality of the patients.
Sujet(s)
Sepsie , Indice APACHE , Adulte , Sujet âgé , Chine , Femelle , Humains , Unités de soins intensifs , Mâle , Adulte d'âge moyen , Pronostic , Courbe ROC , Études rétrospectivesRÉSUMÉ
Grouper, a family of marine fishes, produce distinct vocalizations associated with their reproductive behavior during spawning aggregation. These low frequencies sounds (50-350 Hz) consist of a series of pulses repeated at a variable rate. In this paper, an approach is presented for automatic classification of grouper vocalizations from ambient sounds recorded in situ with fixed hydrophones based on weighted features and sparse classifier. Group sounds were labeled initially by humans for training and testing various feature extraction and classification methods. In the feature extraction phase, four types of features were used to extract features of sounds produced by groupers. Once the sound features were extracted, three types of representative classifiers were applied to categorize the species that produced these sounds. Experimental results showed that the overall percentage of identification using the best combination of the selected feature extractor weighted mel frequency cepstral coefficients and sparse classifier achieved 82.7% accuracy. The proposed algorithm has been implemented in an autonomous platform (wave glider) for real-time detection and classification of group vocalizations.
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Objective: To evaluate the influence of serum procalcitonin in the diagnosis and treatment of pulmonary infection in patients with central nervous system injury. Methods: From October 2014 to February 2017, a retrospective study was performed. A total of 1 852 patients were screened in Department of Intensive Care Unite, First Affiliated Hospital of Sun Yat-sen University.Among them, 173 patients were identified with different kinds of infection. Finally, a total of 42 patients with pulmonary infection were enrolled. The clinical data of patients with pulmonary infection and central nervous system (CNS) injury was collected. A univariate and multivariate regression analysis was performed to study the correlation of serum procalcitonin (PCT) with clinical symptoms and signs of the pulmonary infection, body temperature(T), white blood cell count (WBC), percentage of neutrophils (NEU) and the severity of the pulmonary infection (CPIS). The relationship of serum PCT with type of CNS injury, GCS, and exogenous glucocorticoid was further studied. Results: During the period of pulmonary infection, the peak PCT was 0.83 (0.29, 2.79) µg/L and the CPIS was 5.50 (5.00, 7.00). In 9 of 42 patients, the peak PCT was less than 0.25 µg/L. In 7 of 42 patients, the peak PCT was ranged from 0.25 to 0.5 µg/L. In 12 of 42 patients, PCT was ranged from 0.5 to 2 µg/L. Only 10 patients had a PCT 2-10 µg/L and 4 patients had a PCT more than 10 µg/L. There is no correlation between serum PCT and body temperature, white blood cell, percentage of neutrophils and CPIS. There was no significant differences in patients with PCT<0.5 or ≥0.5 µg/L regarding the body temperature, white blood cell, percentage of neutrophils and CPIS. However, serum PCT in patients with pulmonary infection had independent correlation with the post CNS injury day (ß=0.17, 95% CI (0.02, 0.32), P<0.05). The serum PCT was 1.26 (0.47, 2.7) µg/L and 29.41% patients with a PCT less than 0.5 µg/L within 3 days post CNS injury. Serum PCT level was 0.23 (0.16, 0.39) µg/L, and 77.78% patients with a PCT less than 0.5 µg/L at day 4 to day 7 post-injury. The PCT level was 0.52 (0.33, 1.12) µg/L, and 44.44% patients with a PCT less than 0.5 µg/L at day 8 to day 14. The PCT was 3.26 (2.07, 12.40) µg/L, and no patient with a PCT less than 0.5 µg/L after day 15 post-injury. There were no significant relationship found between serum PCT level and type of the disease and surgery, GCS, and use of exogenous glucocorticoid. Conclusions: Serum PCT had no significant increase and was not able to be used in guiding the antibiotics use in patients with CNS injury and pulmonary infection.
Sujet(s)
Traumatismes du système nerveux , Protéine C-réactive , Calcitonine , Peptide relié au gène de la calcitonine , Humains , Procalcitonine , Précurseurs de protéines , Études rétrospectivesRÉSUMÉ
BACKGROUND: CCK1 (cholecystokinin receptor 1) and protein lysine acetylation were associated with several cancers, respectively. However, whether they are involved in the alternation of gallbladder carcinoma is unknown. AIMS: This study investigated the characteristics of CCK1 and protein lysine acetylation in GBC-SD cells and carcinoma of gallbladder. METHODS: The expression and localization of CCK1 were detected by western blot analysis and indirect immunofluorescence. GBC-SD cells were treated with CCK-8 and CCK-8+CCK1 inhibitor. The protein lysine acetylation from cells, as well as tissues of gallbladder carcinoma, was examined by western blotting. RESULTS: CCK1 receptor was expressed and localized in the GBC-SD cells. The synthetic octapeptide of CCK (CCK-8) could accelerate the lysine acetylation of a subset of proteins in dose-dependent manners in GBC-SD cells. Further investigation demonstrated that the specific inhibitor (CR1409) of CCK1 receptor could attenuate the CCK8-induced increase of protein lysine acetylation. In addition, we revealed that the rise of CCK1 receptor expression is associated with the increase of protein lysine acetylation in tissues from carcinoma of gallbladder. CONCLUSIONS: CCK might regulate protein lysine acetylation via CCK1 receptor.
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Tumeurs de la vésicule biliaire/génétique , Lysine/métabolisme , Récepteur cholécystokinine/métabolisme , Acétylation , Techniques de culture cellulaire , Lignée cellulaire tumorale , Tumeurs de la vésicule biliaire/anatomopathologie , HumainsRÉSUMÉ
OBJECTIVE: To study the mechanism of protection provided by dexmedetomidine against COPD-induced lung injury. METHODS: COPD rat model was determined by measuring lung function, and comparing HE staining between two different groups. We got the lung tissue and cells from the control and COPD groups. The cells were divided into three groups: control group, and blank and drug groups that were from the COPD rats. Cell apoptosis, relative gene expression and TNF-α and IL-1ß from nutrient solution were measured. RESULTS: The TV, PEF, EF50, FEV0.3 and FEV0.3/FVC in COPD group were significantly lower than in control group (1.26±0.17 vs 2.65±0.21; 17.61±0.35 vs 38.55±0.24; 1.20±0.14 vs 1.81±0.06; 2.52±0.28 vs 4.44±0.26; 63.39±0.22 vs 88.45±0.34, p < 0.05, respectively). Cell apoptosis was significantly different in blank and drug groups (21.65±0.86 vs 10.74±0.15; p < 0.05, respectively). The gene expressions of miRNA-146a, p53 and Bcl-2 were significantly downregulated compared with blank group. CONCLUSION: Dexmedetomidine protected COPD-induced lung injury by inhibiting miRNA-146a expression to reduce cell apoptosis (Tab. 1, Fig. 3, Ref. 25).
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Lésion pulmonaire aigüe/prévention et contrôle , Anti-inflammatoires/pharmacologie , Dexmédétomidine/pharmacologie , microARN/génétique , Broncho-pneumopathie chronique obstructive/complications , Lésion pulmonaire aigüe/anatomopathologie , Animaux , Apoptose/génétique , Modèles animaux de maladie humaine , Régulation de l'expression des gènes/effets des médicaments et des substances chimiques , Régulation de l'expression des gènes/génétique , Interleukine-1 bêta , Poumon/métabolisme , Poumon/anatomopathologie , Broncho-pneumopathie chronique obstructive/anatomopathologie , Rats , Rat Sprague-Dawley , Tests de la fonction respiratoire , Facteur de nécrose tumorale alpha/métabolismeRÉSUMÉ
Transient receptor potential cation channel, subfamily M, member 8 (TRPM8) is a nonselective cation channel and a candidate for cold sensation signaling, but the relationship between TRPM8 and diabetes remains unclear. In the present study, we determined the expression levels of TRPM8 messenger RNA (mRNA) and the levels of the TRPM8 protein in the bladder tissue of diabetic rats. We also investigated the correlation between TRPM8 expression and the visceral pain stimulation-related factor, calcitonin gene-related peptide (CGRP) in diabetic rats. The rats were sacrificed 3, 5, 7, and 15 days after streptozotocin injection, and blood was collected from their tail veins to determine the blood glucose levels. Bladder tissue was removed to assess the expression of TRPM8 mRNA by reverse transcription-polymerase chain reaction, and the expression of the TRPM8 protein by western blotting. After administering electrical stimulation (5 V/1 Hz), the expression levels of TRPM8 and CGRP proteins were determined. Our results revealed that the blood glucose level, and TRPM8 mRNA and TRPM8 protein expression levels increased significantly in the diabetic rats. Spinal tissue protein expression levels of both TRPM8 and CGRP also increased significantly following electrical stimulation. This possibly indicates that TRPM8 is closely associated with visceral pain stimulation, and could be an independent prognostic biomarker for diabetes.
Sujet(s)
Diabète expérimental/métabolisme , Canaux cationiques TRPM/génétique , Canaux cationiques TRPM/métabolisme , Douleur viscérale/métabolisme , Animaux , Peptide relié au gène de la calcitonine/génétique , Peptide relié au gène de la calcitonine/métabolisme , Diabète expérimental/génétique , Mâle , ARN messager/génétique , Rats , Rat Sprague-Dawley , Douleur viscérale/génétiqueRÉSUMÉ
Women who carry fragile X mental retardation 1 (FMR1)gene premutation expansions frequently report neurological or endocrine symptoms and prior studies have predominantly focused on questionnaire report of medical issues. Premutation carrier (PMC) women (n = 33) and non-carrier controls (n = 13) were recruited and evaluated by a neurologist, neuropsychologist, and endocrinologist. Blood and skin biopsies were collected for molecular measures. Scales for movement disorders, neuropathy, cognitive function, psychiatric symptoms, sleep, and quality of life were completed. The average age of the women was 51 years (n = 46) and average CGG repeat size was 91 ± 24.9 in the FMR1 PMC women. Seventy percent of the PMC women had an abnormal neurological examination. PMC women had significantly higher scores on the Fragile X-Associated Tremor Ataxia Syndrome (FXTAS) rating scale, more neuropathy, and difficulty with tandem gait compared to controls. Central sensitivity syndromes, a neuroticism profile on the NEO Personality Profile, and sleep disorders were also prevalent. Discrepancies between subject report and examination findings were also seen. This pilot study suggests that women with the FMR1 premutation may have a phenotype that overlaps with that seen in FXTAS. Additional research with larger sample sizes is warranted to better delineate the clinical features.
Sujet(s)
Syndrome du chromosome X fragile/diagnostic , Syndrome du chromosome X fragile/génétique , Études d'associations génétiques , Hétérozygote , Phénotype , Adulte , Sujet âgé , Études cas-témoins , Femelle , Protéine du syndrome X fragile/génétique , Syndrome du chromosome X fragile/physiopathologie , Humains , Adulte d'âge moyen , Mutation , Tests neuropsychologiques , Expansion de trinucléotide répétéRÉSUMÉ
Monocyte interactions with implanted biomaterials can contribute significantly to the ability of a biomaterial to support tissue integration and wound healing, as opposed to a chronic pro-inflammatory foreign body reaction, provided the materials are designed to do so. However, there are few biomaterials available designed to regulate immune cell response with the intention of reducing the pro-inflammatory activation state. Material chemistry is a powerful tool for regulating protein and cell interactions that can be incorporated into surfaces while maintaining desired mechanical properties. The aspects of material chemistry that can support monocyte activation away from a pro-inflammatory state are still poorly understood. Protein adsorption is a key initial event that transforms the surface of a biomedical device into a biological substrate that will govern subsequent cellular interactions. In this study, the chemistry of degradable block polyurethanes, termed degradable polar hydrophobic ionic (D-PHI) polyurethanes, were studied for their unique interactions with bound immunoglobulin G (IgG), a pro-inflammatory protein that supports monocyte-biomaterial interactions. The specific immunological active sites of the polyurethane-adsorbed protein were compared with IgG's adsorbed state on a homopolymeric material with surface chemistry conducive to cell interactions, e.g. tissue culture polystyrene (TCPS). IgG-coated TCPS supported sustained monocyte adhesion and enhanced monocyte spreading, effects not observed with IgG-coated PU. The degradable PU was subsequently shown to reduce the number of exposed IgG-Fab sites following pre-adsorption vs. IgG adsorbed to TCPS, with antibody inhibition experiments demonstrating that Fab-site exposure appears to dominate monocyte-biomaterial interactions. Minor changes in chemical segments within the PU molecular chains were subsequently investigated for their influence on directing IgG interactions towards reducing pro-inflammatory activity. A reduction in chemical heterogeneity within the PU, without significant differences in other material properties known to regulate monocyte response, was shown to increase Fab exposure and subsequently led to monocyte interactions similar to those observed for IgG-coated TCPS. These results infer that reduced IgG-Fab site exposure can be directed by material chemistry to attenuate pro-inflammatory monocyte interactions with biomaterial surfaces, and identify the chemical features of polymeric biomaterial design responsible for this process. STATEMENT OF SIGNIFICANCE: There is currently limited understanding of material design features that can regulate protein-material interactions in order to prevent adverse inflammatory responses to implanted biomaterials. In this paper, monocyte interactions with biomaterials (specifically a block co-polymeric degradable polyurethane [D-PHI] and tissue culture polystyrene [TCPS]) were investigated as a function of their interactions with adsorbed immunoglobulin G (IgG). D-PHI was shown to attenuate IgG-induced monocyte retention and spreading by reducing IgG-Fab site exposure upon adsorption relative to TCPS. Aspects of D-PHI chemistry important in regulating Fab site exposure were determined. This study thus identifies features of biomaterials, using D-PHI as a case study, which can contribute to the development of new immunomodulatory biomaterial design.
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Matières plastiques biodégradables/composition chimique , Matériaux revêtus, biocompatibles/composition chimique , Réaction à corps étranger/immunologie , Immunoglobuline G/composition chimique , Monocytes/immunologie , Polyuréthanes/composition chimique , Adhérence cellulaire/immunologie , Femelle , Humains , Fragments Fab d'immunoglobuline/composition chimique , Mâle , Monocytes/cytologieRÉSUMÉ
Tissue engineering strategies rely on the ability to promote cell proliferation and migration into porous biomaterial constructs, as well as to support specific phenotypic states of the cells in vitro. The present study investigated the use of released factors from monocytes and their derived macrophages (MDM) and the mechanism by which they regulate vascular smooth muscle cell (VSMC) response in a VSMC-monocyte co-culture system within a porous degradable polyurethane (D-PHI) scaffold. VSMCs cultured in monocyte/MDM-conditioned medium (MCM), generated from the culture of monocytes/MDM on D-PHI scaffolds for up to 28 days, similarly affected VSMC contractile marker expression, growth and three-dimensional migration when compared to direct VSMC-monocyte co-culture. Monocyte chemotactic protein-1 (MCP-1) and interleukin-6 (IL-6) were identified as two cytokines present in MCM, at concentrations that have previously been shown to influence VSMC phenotype. VSMCs cultured alone on D-PHI scaffolds and exposed to MCP-1 (5 ng ml(-1)) or IL-6 (1 ng ml(-1)) for 7 days experienced a suppression in contractile marker expression (with MCP-1 or IL-6) and increased growth (with MCP-1) compared to no cytokine medium supplementation. These effects were also observed in VSMC-monocyte co-culture on D-PHI. Neutralization of IL-6, but not MCP-1, was subsequently shown to decrease VSMC growth and enhance calponin expression for VSMC-monocyte co-cultures on D-PHI scaffolds for 7 days, implying that IL-6 mediates VSMC response in monocyte-VSMC co-cultures. This study highlights the use of monocytes and their derived macrophages in conjunction with immunomodulatory biomaterials, such as D-PHI, as agents for regulating VSMC response, and demonstrates the importance of monocyte/MDM-released factors, such as IL-6 in particular, in this process.
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Cytokines/pharmacologie , Macrophages/cytologie , Monocytes/cytologie , Muscles lisses vasculaires/cytologie , Myocytes du muscle lisse/cytologie , Polyuréthanes/pharmacologie , Structures d'échafaudage tissulaires/composition chimique , Technique de Western , Techniques de coculture , ADN/métabolisme , Humains , Protéines et peptides de signalisation intercellulaire/pharmacologie , Myocytes du muscle lisse/effets des médicaments et des substances chimiques , Myocytes du muscle lisse/métabolisme , PorositéRÉSUMÉ
In the present study, we investigate the irradiation-defects hybridized graphene scaffold as one potential building material for the anode of Li-ion batteries. Designating the Wigner V2(2) defect as a representative, we illustrate the interplay of Li atoms with the irradiation defects in graphene scaffolds. We examine the adsorption energetics and diffusion kinetics of Li in the vicinity of a Wigner V2(2) defect using density functional theory calculations. The equilibrium Li adsorption sites at the defect are identified and shown to be energetically preferable to the adsorption sites on pristine (bilayer) graphene. Meanwhile, the minimum energy paths and corresponding energy barriers for Li migration at the defect are determined and computed. We find that, while the defect is shown to exhibit certain trapping effects on Li motions on the graphene surface, it appears to facilitate the interlayer Li diffusion and enhance the charge capacity within its vicinity, because of the reduced interlayer spacing and characteristic symmetry associated with the defect. Our results provide critical assessment for the application of irradiated graphene scaffolds in Li-ion batteries.
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Alimentations électriques , Graphite/composition chimique , Intercalants/composition chimique , Lithium/composition chimique , Diffusion , Ions/composition chimique , CinétiqueRÉSUMÉ
Underlying endothelial dysfunction (EnD) may present in the early stage of ED or psychogenic ED. We retrospectively evaluated 191 ED patients with effective nocturnal penile tumescence and rigidity (NPTR) recording, including detailed medical and psychosexual history, International Index of Erectile Function-5 and vascular parameter. All patients were allocated into psychogenic and organic groups according to the NPTR test. Brachial artery flow-mediated dilation (FMD) was used to diagnose EnD, and ED patients were classified into two groups: non-EnD (FMDî¶10) and EnD (FMD<10). General and vascular parameters were compared between psychogenic and organic groups, and non-EnD and EnD groups with ED were compared in terms of NPTR parameters. In all, 48.7% and 51.3% patients were diagnosed as psychogenic and organic ED, respectively. 73.1% of the psychogenic patients had EnD and 39.8% organic patients had normal endothelial function. In all parameters, only the FMD value showed significant difference between psychogenic and organic ED groups (8.26±2.57 vs 9.16±2.76, P=0.020). No statistical difference was founded in NPTR parameters between non-EnD and EnD groups (P>0.05). In conclusion, NPTR cannot effectively identify the underlying vasculogenic ED from psychogenic ED. Psychogenic causes may cause or aggravate EnD in these ED patients with normal NPTR.
Sujet(s)
Endothélium vasculaire/physiopathologie , Dysfonctionnement érectile/physiopathologie , Érection du pénis/physiologie , Érection du pénis/psychologie , Adolescent , Adulte , Artère brachiale/physiopathologie , Dysfonctionnement érectile/étiologie , Dysfonctionnement érectile/psychologie , Humains , Mâle , Adulte d'âge moyen , Débit sanguin régional/physiologie , Études rétrospectivesRÉSUMÉ
Murine serum inducible kinase (mSnk) was recently cloned and characterized as an early-growth response gene involved in cell proliferation. Here we report the isolation and characterization of its human homologue, named hSnk. Sequence comparison shows that hSnk is highly conserved and its deduced protein sequence shares a significant amino acid identity with mSnk and rSnk proteins, as well as with other polo family kinase gene products. A survey of hSnk expression reveals that while a wide variety of human tissues express a low to moderate level of hSnk transcripts, fetal tissues, testis, and spleen express the most abundant hSnk transcripts. In addition, serum stimulation rapidly induces hSnk expression in fibroblast cells, reaching the peak level of induction within one hour post treatment. Considering that Plk and Prk, two other known human polo-family kinases, control cell cycle checkpoint and cell cycle progression, our current observations suggest that hSnk may also play an important role in cells undergoing rapid cell division or having a high mitotic index.
Sujet(s)
Protéines de Drosophila , Protein kinases/génétique , Protein-Serine-Threonine Kinases/génétique , Adénosine triphosphate/métabolisme , Séquence d'acides aminés , Substitution d'acide aminé , Animaux , Séquence nucléotidique , ADN complémentaire , Humains , Souris , Données de séquences moléculaires , Rats , Similitude de séquences d'acides aminés , Distribution tissulaireRÉSUMÉ
PURPOSE: To study the mechanism of regulation of GSH in HLE-B3 cells expressing alphaA-crystallin (alphaA) and in alphaA knockout mouse lenses. METHODS: GSH levels and maximal rates of GSH synthesis were measured in immortalized, alphaA-transfected HLE-B3 cells containing varying amounts of alphaA. The mRNA and protein for the rate-limiting enzyme for GSH synthesis, gamma-glutamylcysteine synthetase (GCS), were also determined in alphaA- and mock-transfected cells by Northern blot analysis and Western blot analysis of heavy (GCS-HS) and light (GCS-LS) subunits. The effect of absence of alphaA and alphaB on lens GSH concentrations was evaluated in whole lenses of alphaA knockout and alphaB knockout mice as a function of age. GCS-HS mRNA and protein were determined in young, precataractous and cataractous alphaA knockout lenses. RESULTS: GSH levels were significantly higher in HLE-B3 cells expressing alphaA- compared with mock-transfected cells and were correlated positively with alphaA content. Mean rate of GSH synthesis was also higher in alphaA-expressing cells than in mock controls (0.84 vs. 0.61 nmol. min(-1) per mg protein, respectively). GCS-HS mRNA and GCS-LS mRNA were approximately twofold higher in alphaA-expressing cells, whereas the heavy and light GCS subunit proteins increased by 80% to 100%. In alphaA(-/-) mouse lenses, GSH level was not different from that of wild type up to 2 months from birth, after which it dropped to approximately 50% of controls. On the other hand, GCS-HS and GCS-LS proteins showed a significant decrease before cataract formation as early as 15 days after birth. GSH level in cataract-free alphaB(-/-) lenses was similar to that of wild type for up to 14 months. CONCLUSIONS: Expression of alphaA caused an increase in cellular GSH, in part, because of an increase in mRNA and protein of both GCS subunits. GSH levels decreased with increasing age in cataractous alphaA(-/-) lenses but not in the noncataractous alphaB(-/-) lenses. It is suggested that neonatal precataractous lenses (with normal GSH and decreased GCS) may maintain their GSH level by other compensatory mechanisms such as increased GSH transport.
Sujet(s)
Cristallines/métabolisme , Cellules épithéliales/métabolisme , Glutathion/métabolisme , Cristallin/métabolisme , Animaux , Technique de Northern , Technique de Western , Lignée cellulaire , Cristallines/génétique , Expression des gènes , Glutamate-cysteine ligase/génétique , Glutamate-cysteine ligase/métabolisme , Humains , Nourrisson , Souris , Souris knockout , ARN messager/métabolisme , TransfectionRÉSUMÉ
Activating mutations of RAS are thought to be early events in the evolution of thyroid follicular neoplasms. We used a doxycycline-inducible expression system to explore the acute effects of H-RAS12 on genomic stability in thyroid PCCL3 cells. At 2-3 days (first or second cell cycle) there was a significant increase in the frequency of micronucleation. Treatment of cells with YVAD-CHO inhibited RAS-induced apoptosis, but had no effect on micronucleation. The effects of H-RAS(V12) were mediated by activation of MAPK, as treatment with PD98059 at concentrations verified to selectively inhibit MEK1 reduced the frequency of prevalence of cells with micronuclei. In addition, doxycycline-inducible expression of a constitutively active MEK1, but not of a mutant RAC1, mimicked the effects of H-RAS(V12). The effects of H-RAS(V12) on genome destabilization were apparent even though the sequence of p53 in PCCL3 cells was confirmed to be wild-type. Acute activation of H-RAS(V12) evoked a proportional increase in both CREST negative and CREST positive micronuclei, indicating that both clastogenic and aneugenic effects were involved. H-RAS(V12) and activated MEK1 also induced centrosome amplification, and chromosome misalignment. Evidence that acute expression of constitutively activated RAS destabilizes the genome of PCCL3 cells is consistent with a mode of tumor initiation in which this oncogene promotes phenotypic progression by predisposing to large scale genomic abnormalities.