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The precise synthesis of poly(thioester)s with diverse structures is still a significant challenge in the polymeric materials field. Herein, we report a novel approach to the synthesis of well-defined poly(thioester)s by the controlled alternating copolymerization of cyclic thioanhydrides and episulfides induced by simple organic ammonium salts. Both the cation and anion have strong effects on the copolymerization. [PPN]OAc ([PPN]=bis(triphenylphosphine)iminium) with a bulky cation was proven to be efficient in initiating this polymerization, yielding poly(thioester)s with a completely alternating structure, controlled molecular weight, and narrow polydispersity. The poly(thioester) obtained from succinic thioanhydride and propylene sulfide is a typical semicrystalline material, possessing a high refractive index of up to 1.78. Because it uses readily available monomers, this method is expected to open up a new route to poly(thioester)s with diverse structures and properties.
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Despite advances in therapeutic strategies, heart failure-associated mortality rates remain high. Thus, understanding the pathophysiological molecular mechanisms involved in the remodeling process is essential for developing new and effective therapies. LRRs are present various prokaryotic and eukaryotic proteins and important for the innate immune system via regulating protein-protein interactions. LRRC25 is a member of leucine-rich repeat (LRR)-containing protein family. LRRC25 has been shown to negatively modulate nuclear factor κB (NF-κB) activation, a crucial factor related to cardiac hypertrophy. Our aim was to explore the effects of LRRC25 on cardiac hypertrophy. In the present study, LRRC25 levels were decreased in human and mouse hypertrophied hearts. LRRC25 knockout exacerbated cardiac hypertrophy responding to pressure overloading or angiotensin II (Ang II) stimulation. Deletion of LRRC25 accelerated cardiac dysfunction and fibrosis in mice subjected to aortic banding (AB). LRRC25 ablation induced a strong increase in the transcription of both hypertrophy (ANP, BNP, and ß-MHC) and fibrosis associated molecules (col1, col3a1, α-SMA and fibronectin). In addition, the expression of transforming growth factor-ß1 (TGF-ß1), and its down-streaming signals of phosphorylated Smad2/3, was markedly induced by LRRC25 deficiency. LRRC25-knockout mice showed a significantly enhanced inflammation in response to AB surgery by promoting the activation of NF-κB signaling pathway. In mouse cardiomyocytes, LRRC25 deficiency markedly elevated TGF-ß1 and NF-κB activation stimulated by Ang II. Treatment with a combination of TGF-ß1 or NF-κB inhibitor abolished the effects of LRRC25-knockout on the promotion of cardiac hypertrophy in vitro. Together, our study identified LRRC25 as a critical molecular switch whose down-regulation resulted in cardiac hypertrophy in a TGF-ß1- and NF-κB-dependent manner.
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Cardiomegalia/patologia , Fibrose/etiologia , Inflamação/etiologia , Proteínas de Membrana/fisiologia , Fator de Crescimento Transformador beta1/farmacologia , Animais , Cardiomegalia/etiologia , Fibrose/induzido quimicamente , Técnicas de Inativação de Genes , Humanos , Técnicas In Vitro , Inflamação/induzido quimicamente , Proteínas de Membrana/deficiência , Proteínas de Membrana/genética , Camundongos , Miócitos Cardíacos/metabolismo , NF-kappa B/metabolismo , NF-kappa B/farmacologia , Fator de Crescimento Transformador beta1/metabolismoRESUMO
AIM: Cryoglobulinaemic glomerulonephritis related to hepatitis B virus (HBV) infection has been rarely reported. The aim of this study was to investigate the clinical features, renal biopsy findings in patients with HBV-associated cryoglobulinaemia. METHODS: Twelve patients with HBV-associated cryoglobulinaemia were identified in this study. The demographic, clinical, pathological characteristics, treatment and follow-up data were analyzed. RESULTS: Renal involvement was characterized by nephrotic range proteinuria with microscopic haematuria in all patients, and impaired renal function in nine patients (75%). Purpuric rash was the main extrarenal manifestation (58.3%). Type II cryoglobulinaemia was presented in three patients and type III in nine patients. Hypocomplementaemia and positive of rheumatoid factors were present in all patients. Membranoproliferative glomerulonephritis (MPGN) was observed in all kidney specimens. Seven patients also had evidence of prominent cryoglobulins thrombi on renal biopsy, but only three patients had HBV antigen deposits in renal tissues. Antiviral and steroids or immunosuppressive agents have been used in most of patients. During follow-up, two patients died, and four reaching end-stage renal disease; three patients had complete remission, and three patients had renal function improved after therapy. CONCLUSIONS: Nephrotic syndrome with haematuria and renal insufficiency are the main clinical manifestation; cryoglobulinaemic glomerulonephritis are the main renal lesion in patients with HBV-Associated cryoglobulinaemia; half of patients have poor outcome even with antiviral and immunosuppressive therapy. The results of this study indicate that cryoglobulins should be detected in hepatitis B virus-Associated nephropathy in endemic area.
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Crioglobulinemia/complicações , Glomerulonefrite/diagnóstico , Glomerulonefrite/etiologia , Hepatite B/complicações , Adulto , Idoso , Antivirais/uso terapêutico , Crioglobulinemia/patologia , Crioglobulinemia/virologia , Feminino , Glomerulonefrite/terapia , Hepatite B/patologia , Hepatite B/terapia , Humanos , Imunossupressores/uso terapêutico , Masculino , Pessoa de Meia-Idade , Avaliação de Resultados em Cuidados de Saúde , Estudos RetrospectivosRESUMO
Serum phospholipase A2 receptor antibodies (SAbs) and glomerular phospholipase A2 receptor antigen (GAg) deposits have been observed in idiopathic membranous nephropathy (IMN). However, the clinical application of these two biomarkers, particularly GAg deposition, needs to be further evaluated. We measured SAb concentration by ELISA and GAg deposition by immunofluorescence in 572 patients with biopsy-proven IMN. Overall, 68.5% of patients (392 of 572) had detectable SAb (SAb+), and 98.7% of patients who were SAb+ (387 of 392) and 70.6% of patients who were SAb- (127 of 180) had GAg deposition (GAg+). Compared with patients who were SAb-/GAg+, patients who were SAb+/GAg+ exhibited higher levels of proteinuria (P<0.001) and a lower chance of proteinuria remission (P<0.001). In 52 patients who underwent repeat biopsies, patients who did not achieve remission had a higher SAb+ rate on the first biopsy than patients who went into remission (P=0.001). Furthermore, SAb+ levels persisted in patients who did not achieve remission but significantly decreased in patients who achieved remission by the second biopsy. Patients who did not achieve remission also had a higher GAg+ rate on the first biopsy than patients who achieved remission (P<0.01). Sustained GAg+ deposits correlated with disease relapse. In conclusion, combining the measurements of SAb levels and detection of GAg deposition may provide additional information regarding diagnoses, treatment response, and disease relapse in patients with IMN.
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Autoanticorpos/imunologia , Glomerulonefrite Membranosa/imunologia , Glomérulos Renais/metabolismo , Receptores da Fosfolipase A2/imunologia , Receptores da Fosfolipase A2/metabolismo , Adulto , Autoanticorpos/sangue , Biópsia , Feminino , Glomerulonefrite Membranosa/sangue , Glomerulonefrite Membranosa/patologia , Humanos , Glomérulos Renais/patologia , Masculino , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
OBJECTIVE: To investigate the correlation of the single nucleotide polymorphism (SNP) rs4880 of the superoxide dismutase 2 (SOD2) gene with the risk of male infertility. METHODS: This caseîcontrol study included 519 male patients with idiopathic infertility (aged 19ï¼40 ï¼»28.93±4.93ï¼½ years) in the case group and 338 fertile men (aged 19ï¼40 ï¼»28.40±4.25ï¼½ years) in the control group. We collected the clinical data, genotyped the SNP rs4880 of the SOD2 gene by Sequenom Mass Array, and analyzed the association of different genotypes with male infertility using the logistic regression model. RESULTS: Statically significant differences were observed between the case and control groups in the level of follicleîstimulating hormone (FSH) (ï¼»4.72±2.51ï¼½ vs ï¼»15.65±17.24ï¼½ U/L, P< 0.01), the percentage of progressively mobile sperm (ï¼»9.12±13.5ï¼½ vs ï¼»41.95±9.03ï¼½%, P< 0.01), and sperm concentration (ï¼»12.95±24.38ï¼½ vs ï¼»72.88±45.60ï¼½ ×106/ml, P< 0.01), but not in other parameters. No correlation was found between male infertility and the heterozygous genotype TC (OR = 0.90, 95% CI: 0.65ï¼1.25, P = 0.516) or the homozygous genotype CC (OR=1.49, 95% CI: 0.38ï¼5.81, P = 0.566) as compared with the wild genotype TT, and similar results were obtained in the analysis of the subgroups. CONCLUSIONS: The SNP rs4880 of the SOD2 gene was not correlated with male infertility, which, however, is to be supported by further studies with larger samples from more areas.
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Infertilidade Masculina/genética , Polimorfismo de Nucleotídeo Único , Superóxido Dismutase/genética , Adulto , Estudos de Casos e Controles , Hormônio Foliculoestimulante/sangue , Predisposição Genética para Doença , Genótipo , Heterozigoto , Humanos , Modelos Logísticos , Masculino , Nucleotídeos/genética , Motilidade dos Espermatozoides , Adulto JovemRESUMO
BACKGROUND: The association between psoriasis and membranous nephropathy (MN) remains largely unclear. We examined the prevalence of serum PLA2R antibody and characterized the expression of PLA2R and THSD7A in glomeruli in patients with MN and psoriasis. METHODS: A total of 24 patients with MN without evidence of a secondary cause except psoriasis were enrolled. The clinical and pathological features were retrospectively analyzed. Serum anti-PLA2R antibody was measured using IFA Mosaic. Renal tissue samples stored in the laboratory bio-bank were used for PLA2R staining under immunofluorescence microscopy and THSD7A immunohistochemical analysis. RESULTS: Twenty-four patients (21 male and 3 female) with a mean age of 43.6 ± 15.7 years old were enrolled. Serum anti-PLA2R antibody was positive in 7 patients, which was significantly lower than the positivity observed in idiopathic MN (29.2% vs. 81.7%, P < 0.001). Glomerular PLA2R staining was positive in 7 patients with positive serum anti-PLA2R antibody. THSD7A staining was negative in all 24 patients. During the follow-up visits, 13 patients with negative serum PLA2R antibody achieved CR. In contrast, CR was only achieved in 1 patient with positive serum PLA2R antibody, PR was achieved in 2 patients. CONCLUSIONS: The prevalence of serum anti-PLA2R antibody and glomerular expression of PLA2R was significantly lower in patients with psoriasis and MN than in those with idiopathic MN, and THSD7A staining was negative, suggesting that MN is associated with psoriasis in the majority of patients. However, idiopathic MN might also accompany psoriasis in a minority of psoriatic patients with positive serum anti-PLA2R antibody.
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Autoanticorpos/sangue , Glomerulonefrite Membranosa/metabolismo , Glomérulos Renais/metabolismo , Psoríase/sangue , Receptores da Fosfolipase A2/imunologia , Receptores da Fosfolipase A2/metabolismo , Adolescente , Adulto , Idoso , Feminino , Glomerulonefrite Membranosa/complicações , Glomerulonefrite Membranosa/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-Idade , Proteinúria/etiologia , Psoríase/complicações , Psoríase/tratamento farmacológico , Estudos Retrospectivos , Trombospondinas/metabolismo , Resultado do Tratamento , Adulto JovemRESUMO
The mechanism by which glucocorticoids alleviate renal inflammatory disorders remains incompletely understood. Here, we report that the efficacy of glucocorticoids in ameliorating FSGS depends on the capacity to expand myeloid-derived suppressor cells (MDSCs). After glucocorticoid treatment, the frequency of CD11b(+)HLA-DR(-)CD14(-)CD15(+) MDSCs in peripheral blood rapidly increased in patients with glucocorticoid-sensitive FSGS but remained unchanged in patients with glucocorticoid-resistant FSGS. The frequency of CD11b(+)Gr-1(+) MDSCs in mouse peripheral blood, bone marrow, spleen, kidney-draining lymph nodes (KDLNs), and kidney also increased after glucocorticoid treatment. The induced MDSCs from glucocorticoid-treated mice strongly suppressed T cells, dendritic cells, and macrophages but induced regulatory T cells in spleen, KDLNs, and kidney. Moreover, glucocorticoid treatment suppressed doxorubicin-induced T cell proliferation, dendritic cell and macrophage infiltration, and proinflammatory cytokine production, whereas this protective effect was largely abolished by depleting MDSCs using anti-Gr-1 antibody. Finally, the adoptive transfer of induced MDSCs into the doxorubicin-treated mice not only confirmed the protective role of MDSCs in doxorubicin-induced renal injury but also showed that the transferred MDSCs rapidly migrated into the lymphocyte-accumulating organs, such as the spleen and KDLNs, where they suppressed T cell proliferation. Taken together, these results demonstrate that glucocorticoid treatment ameliorates FSGS by expanding functional MDSCs and that this rapid elevation of MDSCs in peripheral blood may serve as an indicator for predicting the efficacy of glucocorticoid treatment.
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Células Dendríticas/efeitos dos fármacos , Glomerulosclerose Segmentar e Focal/sangue , Glomerulosclerose Segmentar e Focal/tratamento farmacológico , Glucocorticoides/farmacologia , Células Mieloides/efeitos dos fármacos , Linfócitos T Reguladores/efeitos dos fármacos , Adolescente , Transferência Adotiva , Adulto , Animais , Antígenos Ly/análise , Antígeno CD11b/análise , Contagem de Células , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Doxorrubicina/farmacologia , Feminino , Glomerulosclerose Segmentar e Focal/patologia , Antígenos HLA-DR/análise , Humanos , Rim/patologia , Antígenos CD15/análise , Receptores de Lipopolissacarídeos/análise , Linfonodos/patologia , Macrófagos/efeitos dos fármacos , Masculino , Camundongos , Pessoa de Meia-Idade , Células Mieloides/química , Receptores de Quimiocinas/análise , Adulto JovemRESUMO
To investigate the role of mast cells (MCs) renal infiltration in the progression of human anti-GBM nephritis, 38 patients diagnosed with anti-GBM nephritis were enrolled. Renal biopsies were performed. Immunohistochemistry was conducted to detect MCs in renal tissues. Patients were divided into group 1 (MCs <50 mm(-2), n = 18) and group 2 (MCs ≥50 mm(-2), n = 20) according to the infiltrating renal MC count. The clinical-pathological indices were compared. And, correlation between MCs and the clinical-pathological indices was analyzed. Patients of group 2 had more severe renal dysfunctions, expressed as higher levels of serum creatinine (SCr 8.95 ± 3.66 vs. 4.75 ± 2.73 mg/dL, p < 0.001), urine retinol-binding protein (RBP 29.8 ± 13.9 vs. 15.7 ± 11.5 mg/dL, p = 0.005), and lower urinary osmotic pressure. Pathologically, patients of group 2 had a higher percentage of fibrous/fibrocellular crescents (66.7 ± 21.9 vs. 47.0 ± 33.6%, p = 0.037) but a lower percentage of cellular crescents. More CD8 (268 mm(-2) vs. 180 mm(-2), p = 0.045) and CD68 (268 mm(-2) vs. 180 mm(-2), p = 0.045) positive cells infiltrating the interstitium were observed in group 2. Furthermore, renal MCs correlated significantly with the total number of crescents and the tubular interstitial CD8 and CD68 positive cells. And, the number of MCs was associated with the histological types. The renal function was significantly different between the two groups at presentation. However, at 3 and 6 month follow-up, the patient outcome was associated with the histological types. Our study showed that MC infiltrations were associated with chronic lesions in anti-GBM nephritis and may be involved in the loss of renal function with pathological changes.
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Doença Antimembrana Basal Glomerular/patologia , Rim/patologia , Mastócitos/citologia , Proteínas de Ligação ao Retinol/urina , Adulto , Doença Crônica , Creatinina/sangue , Feminino , Fibrose , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Pressão Osmótica , Adulto JovemRESUMO
OBJECTIVE: To determine the correlation of the CYP1A1 (rs4646422) gene polymorphisms with male infertility in the Chinese Han population. METHODS: Using the Mass ARRAY iPLEX GOLD technique, we conducted a case-control study on theCYPlA1 (rs4646422) gene polymorphisms in 636 infertile males aged 21-49 years (case group) and 442 normal healthy men aged 23-47 years (control group) of the Chinese Han population. We analyzed the genotypes and allele frequencies in the two groups ofsubjects with the SPSS 20.0 software. RESULTS: Compared with the wild homozygous genotype GG, the heterozygous genotype AG (OR = 1.06, 95% CI 0.81-1.38) and homozygous genotype AA (OR = 1.11, 95% CI 0.56-2.21) showed no correlation with male infertility, nor did the mutant allele A (OR = 1.06, 95% CI 0.85-1.32) in comparison with the wild allele G. CONCLUSION: The CYP1A1 (rs4646422) gene polymorphisms might not be correlated with male infertility in the Chinese Han population.
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Citocromo P-450 CYP1A1/genética , Infertilidade Masculina/genética , Polimorfismo Genético , Adulto , Alelos , Estudos de Casos e Controles , China , Frequência do Gene , Genótipo , Homozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
The auto-test equipment for relative spectral response of charge coupled devices (CCD) was designed and realized. The equipment covered the range of 400 to 950 nm. Firstly, testing theory of relative spectral response for detectors was introduced. Secondly, taking the high sensitive and broad spectral scientific spectrometer-QE65000 as reference, auto-test equipment for relative spectral response was built up by direct comparison method in the same radiation field. Uncertainty analysis showed that the maximum uncertainty of the equipment was less than 6.21%. This auto-test equipment can be used in the CCD assessment and its photoelectrical parameters testing.
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Alport syndrome (AS) is an inherited glomerular basement membrane (GBM) disease leading to end-stage renal disease (ESRD). X-linked AS (XLAS) is caused by pathogenic variants in the COL4A5 gene. Many pathogenic variants causing AS have been detected, but the genetic modifications and pathological alterations leading to ESRD have not been fully characterized. In this study, a novel frameshift variant c.980_983del ATGG in the exon 17 of the COL4A5 gene detected in a patient with XLAS was introduced into a mouse model in by CRISPR/Cas9 system. Through biochemical urinalysis, histopathology, immunofluorescence, and transmission electron microscopy (TEM) detection, the clinical manifestations and pathological alterations of Del-ATGG mice were characterized. From 16 weeks of age, obvious proteinuria was observed and TEM showed typical alterations of XLAS. The pathological changes included glomerular atrophy, increased monocytes in renal interstitial, and the absence of type IV collagen α5. The expression of Col4a5 was significantly decreased in Del-ATGG mouse model. Transcriptomic analysis showed that differentially expressed genes (DEGs) accounted for 17.45% (4,188/24003) of all genes. GO terms indicated that the functions of identified DEGs were associated with cell adhesion, migration, and proliferation, while KEGG terms found enhanced the degradation of ECM, amino acid metabolism, helper T-cell differentiation, various receptor interactions, and several important pathways such as chemokine signaling pathway, NF-kappa B signaling pathway, JAK-STAT signaling pathway. In conclusion, a mouse model with a frameshift variant in the Col4a5 gene has been generated to demonstrate the biochemical, histological, and pathogenic alterations related to AS. Further gene expression profiling and transcriptomic analysis revealed DEGs and enriched pathways potentially related to the disease progression of AS. This Del-ATGG mouse model could be used to further define the genetic modifiers and potential therapeutic targets for XLAS treatment.
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In order to realize the irradiance calibration of SBUV-ICCD (solar blind ultraviolent Intensified change-coupled device) with dynamic range reaching 120 dB, a light source with long dynamic range was designed and realized. Firstly, the irradiance dynamic range was estimated. Then using deuterium lamp, integrating sphere, precise stop and rail, an ultraviolent light source was developed, which has fixed structure of spectrum, but the irradiance can change continuously in long range. At last the light source's performance was tested. The result shows that the irradiance between 0.278 and 2.8 x 10(-7) microW x cm(-2) was covered, and the stability was 0.93%/3 h. So the demand of calibration of irradiance was satisfied. It will help for measuring the surface uniformity of detector and the calibration of imaging systems.
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OBJECTIVE: To investigate the application of high offset femoral stem prosthesis in primary total hip arthroplasty. METHODS: From January 2015 to June 2017, 51 patients with unilateral hip diseases who underwent primary total hip arthroplasty with Corail high offset femoral stem prosthesis(KHO type) were selected for retrospective study, including 20 females and 31 males;the age ranged from 21 to 71 years old with an average of(50.8±13.3) years old. The abduction arm, femoral offset, acetabular offset and the length of lower limbs were measured on the positive X-ray film of hip joint after operation. Harris scores before and after operation and related complications were recorded, and the stability of prosthesis was analyzed. RESULTS: The femoral offset, combined offset and abduction arm of the affected side were significantly greater than those of the healthy side(P<0.05). There was no significant difference in acetabular offset between the affected side and the healthy side (P>0.05). The femoral offset of 17 hips (33.3%) was reconstructed normally, of which 15 cases (88.2%) had equal length of both lower limbs. The femoral offset of 34 hips (66.7%) was greater than that of the healthy side, and 34 cases (100%) had equal length of both lower limbs. All 51 patients were followed up for(42.3±7.3) months. The Harris score increased from 38.0±7.6 before operation to 92.1±3.1 at the final follow-up(P<0.001). CONCLUSION: Although the high offset Corail prosthesis can not normally reconstruct the femoral offset in unilateral primary total hip arthroplasty, it does not affect the reconstruction of the length of lower limbs and the stability of the prosthesis.
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Artroplastia de Quadril , Prótese de Quadril , Adulto , Idoso , Feminino , Seguimentos , Articulação do Quadril/cirurgia , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Resultado do Tratamento , Adulto JovemRESUMO
An intensification of the "electrophile-nucleophile" synergistic effect was achieved in a microreactor for the coupling reaction of CO2 and epoxides mediated by the binary Al complex/ternary ammonium salt catalyst system. The microreactor technology is proven to be a powerful tool for the preparation of cyclic carbonates with an improved reaction rate and a wide substrate scope.
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Dyschromatosis symmetrica hereditaria (DSH) is a rare autosomal dominant pigmentary genodermatosis, which is characterized by a mixture of hyperpigmented and hypopigmented macules on the dorsal of the hands and feet, and on the face presented like freckle. Identification of RNA-specific adenosine deaminase 1 (ADAR1) gene results in DSH. This study was mainly to explore the pathogenic mutation of ADAR1 gene and provide genetics counselling and prenatal diagnostic testing for childbearing individuals.Mutational analysis of ADAR1 gene was performed by polymerase chain reaction (PCR) and electrophoretic separation of PCR products by 1.5% agarose gel electrophoresis. The coding exons and intron/exon flanking regions followed by bidirectional sequencing was performed on all participants. In this study, we found that a 28 year-old male patient harbouring a deleterious substitution of Leu1052Pro in the ADAR1 gene in a typical DSH family. His mother suffered from the DSH also owns the same mutation. This mutation, however, is not identified in the unaffected members in this family and those 200 normal controls. In summary, this new mutation Leu1052Pro reported here is pathogenic and detrimental for DSH. Our finding not only enriches mutation database and contributes to dissecting further the correlation between mutation position and phenotypical features of DSH, but also provides genetics counselling and prenatal diagnostic testing for childbearing couple.
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Adenosina Desaminase/genética , Predisposição Genética para Doença , Transtornos da Pigmentação/congênito , Pigmentação/genética , Proteínas de Ligação a RNA/genética , Adulto , Análise Mutacional de DNA , Éxons/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mutação de Sentido Incorreto , Transtornos da Pigmentação/genética , Transtornos da Pigmentação/fisiopatologiaRESUMO
Alport syndrome (AS) is a clinically and genetically heterogeneous, progressive nephropathy caused by mutations in COL4A3, COL4A4, and COL4A5, which encode type IV collagen. The large sizes of these genes and the absence of mutation hot spots have complicated mutational analysis by routine polymerase chain reaction (PCR)-based approaches. Here, in order to design a rapid and effective method for the genetic diagnosis of AS, we developed a strategy by utilizing targeted capture associated with next-generation sequencing (NGS) to analyze COL4A3, COL4A4, and COL4A5 simultaneously in 20 AS patients. All the coding exons and flanking sequences of COL4A3, COL4A4, and COL4A5 from the probands were captured followed by HiSeq 2500 sequencing. Candidate mutations were validated by classic Sanger sequencing and quantitative (q)PCR. Sixteen patients (16/20, 75%) showed X-linked inheritance, and four patients (4/20, 20%) showed autosomal recessive inheritance. None of the individuals had autosomal-dominant AS. Fifteen novel mutations, 6 known mutations, and 2 novel fragment deletions were detected by targeted capture and NGS. Of these novel mutations, 12, 3, and 2 mutations were detected in COL4A5, COL4A4, and COL4A3, respectively. A comparison of the clinical manifestations caused by different types of mutations in COL4A5 suggested that nonsense mutations and glycine substitution by an acidic amino acid are more severe than the other missense mutations. Pathogenic mutations were detected in 20 patients. These novel mutations can expand the genotypic spectrum of AS. Our results demonstrated that targeted capture and NGS technology are effective in the genetic diagnosis of AS.
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Povo Asiático/genética , Autoantígenos/genética , Colágeno Tipo IV/genética , Mutação , Nefrite Hereditária/genética , Adolescente , Adulto , Criança , Pré-Escolar , China , Colágeno Tipo IV/deficiência , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Deleção de Sequência , Adulto JovemRESUMO
BACKGROUND: The interaction between activated microglia and T lymphocytes can yield abundant pro-inflammatory cytokines. Our previous study proved that thymus immune tolerance could alleviate the inflammatory response. This study aimed to investigate whether intrathymic injection of myelin basic protein (MBP) in mice could suppress the inflammatory response after co-culture of T lymphocytes and BV-2 microglia cells. METHODS: Totally, 72 male C57BL/6 mice were randomly assigned to three groups (n = 24 in each): Group A: intrathymic injection of 100 µl MBP (1 mg/ml); Group B: intrathymic injection of 100 µl phosphate-buffered saline (PBS); and Group C: sham operation group. Every eight mice in each group were sacrificed to obtain the spleen at postoperative days 3, 7, and 14, respectively. T lymphocytes those were extracted and purified from the spleens were then co-cultured with activated BV-2 microglia cells at a proportion of 1:2 in the medium containing MBP for 3 days. After identified the T lymphocytes by CD3, surface antigens of T lymphocytes (CD4, CD8, CD152, and CD154) and BV-2 microglia cells (CD45 and CD54) were detected by flow cytometry. The expressions of pro-inflammatory factors of BV-2 microglia cells (interleukin [IL]-1ß, tumor necrosis factor-α [TNF-α], and inducible nitric oxide synthase [iNOS]) were detected by quantitative real-time polymerase chain reaction (PCR). One-way analysis of variance (ANOVA) and the least significant difference test were used for data analysis. RESULTS: The levels of CD152 in Group A showed an upward trend from the 3rd to 7th day, with a downward trend from the 7th to 14th day (20.12 ± 0.71%, 30.71 ± 1.14%, 13.50 ± 0.71% at postoperative days 3, 7, and 14, respectively, P < 0.05). The levels of CD154 in Group A showed a downward trend from the 3rd to 7th day, with an upward trend from the 7th to 14th day (10.00 ± 0.23%, 5.28 ± 0.69%, 14.67 ± 2.71% at postoperative days 3, 7, and 14, respectively, P < 0.05). The ratio of CD4+/CD8 + T in Group A showed a downward trend from the 3rd to 7th day, with the minimum at postoperative day 7, then an upward trend from the 7th to 14th day (P < 0.05). Meanwhile, the levels of CD45 and CD54 in Group A were found as the same trend as the ratio of CD4+/CD8 + T (CD45: 83.39 ± 2.56%, 82.74 ± 2.09%, 87.56 ± 2.11%; CD54: 3.80 ± 0.24%, 0.94 ± 0.40%, 3.41 ± 0.33% at postoperative days 3, 7, and 14, respectively, P < 0.05). The expressions of TNF-α, IL-1ß, and iNOS in Group A were significantly lower than those in Groups B and C, and the values at postoperative day 7 were the lowest compared with those at postoperative days 3 and 14 (P < 0.05). No significant difference was found between Groups B and C. CONCLUSIONS: Intrathymic injection of MBP could suppress the immune reaction that might reduce the secondary immune injury of brain tissue induced by an inflammatory response.
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Anti-Inflamatórios/farmacologia , Microglia/imunologia , Proteína Básica da Mielina/farmacologia , Linfócitos T/imunologia , Animais , Antígenos de Superfície/análise , Lesões Encefálicas Traumáticas/tratamento farmacológico , Relação CD4-CD8 , Técnicas de Cocultura , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteína Básica da Mielina/administração & dosagemRESUMO
BK virus infection accompanied with plasma cell-rich infiltrates is a dilemma in renal transplant recipients. One young female patient diagnosed as BK virus-associated nephropathy with plasma cell-rich infiltrates at 16 months after renal transplant was treated with bortezomib and a sequential immuno-suppressive protocol of tacrolimus combined with leflunomide. After a short period of reduction, her serum creatinine increased slowly with stable BK viruria. The patient underwent repeat biopsy. The histologic changes showed a decrease in plasma cells and CD20+ cells in the allograft, but the other mononuclear cells showed no difference from the first biopsy. The immunosuppressive protocol was converted to tacrolimus combined with enteric-coated mycophenolate sodium. Her serum creatinine decreased gradually during 6 months of follow-up. We speculate that bortezomib can be used in BK virus-associated nephropathy accompanied with plasma cell-rich infiltrates, and this effect might be mediated through a decrease of plasma cells and CD20+ cells in the allograft. The dosage and time of therapy need to be explored in the future; additional studies of large samples are needed.
Assuntos
Antineoplásicos/uso terapêutico , Vírus BK , Bortezomib/uso terapêutico , Nefropatias/tratamento farmacológico , Nefropatias/patologia , Transplante de Rim , Plasmócitos/patologia , Infecções por Polyomavirus/tratamento farmacológico , Infecções por Polyomavirus/patologia , Infecções Tumorais por Vírus/tratamento farmacológico , Infecções Tumorais por Vírus/patologia , Adulto , Feminino , Humanos , Isoxazóis/uso terapêutico , Leflunomida , Complicações Pós-Operatórias/tratamento farmacológico , Tacrolimo/uso terapêuticoRESUMO
The current study was performed to investigate the effect of aminoguanidine (AG) on spinal cord injury (SCI) in rat. AG (75, 150 and 300mg/kg, i.p. respectively ) was administered to rats immediately following SCI. It was found that AG (150mg/kg) significantly reduced spinal cord water content and improved motor function, however, AG at the doses of 75 and 300mg/kg had no effect. Compared to SCI group without treatment, AG at the dosage of 150mg/kg induced a reduction in the permeability of blood-spinal cord barrier (BSCB) after injury 48h (from 59.8+/-5.5microl/g to 39.8+/-3.8microl/g), a 38% decrease of Malondialdehyde (MDA) values and a 1-fold increase of the Glutathione (GSH) levels at 12h after SCI. And the expression of inducible nitric oxide synthase (iNOS) protein reached a peak at 24h after injury, which was significantly attenuated by treatment with AG (150mg/kg). In addition, the expression of AQP4 protein was down-regulated by the treatment of AG (150mg/kg) at 24h after SCI, and the changes still lasted at 48h after injury. Our results indicated that AG could induce spinal cord edema clearance and improve motor function, which could be correlated with antioxidative property, the down-regulation of iNOS and AQP4 protein expression after SCI.