RESUMO
CTLs play important roles in host immune responses to tumors. CD4 CTLs are characterized by their ability to secrete cytotoxic effector molecules, such as granzyme B and perforin, and kill target cells in a MHC class II-restricted manner. However, the cell surface markers of CD4 CTLs remain unknown, which hinders their separation and research on their function. In this study, we performed a bioinformatics analysis and experimental validation that revealed that G protein-coupled receptor 56 (GPR56) is a cell surface marker that can be used to characterize CD4 CTLs. We found that GPR56 and granzyme B were coexpressed in extremely high levels in human peripheral blood T cells, and that anti-GPR56 stimulation significantly upregulated the expression of granzyme B in both CD4+GPR56+ and CD8+GPR56+ T cells. These findings suggest that GPR56 expression and the GPR56 signaling pathway could contribute directly to the toxic function of either CD4+ or CD8+ T cells. We also used GPR56 as a biomarker to investigate the clinical significance of CD4 CTLs. GPR56+ T cell levels were increased in patients with lung cancer, and GPR56 expression was significantly correlated with lung cancer progression. A further analysis revealed an increase in exhausted cell states in lung cancer patients because of upregulation of programmed cell death protein 1 expression in GPR56+ T cells. The findings of this study suggest that GPR56 characterizes the cytotoxic states of either CD4+ or CD8+ T cells.
Assuntos
Linfócitos T CD4-Positivos , Neoplasias Pulmonares , Humanos , Granzimas/metabolismo , Linfócitos T Citotóxicos , Receptores Acoplados a Proteínas G/metabolismo , Neoplasias Pulmonares/metabolismoRESUMO
Although the roles of E proteins and inhibitors of DNA-binding (Id) in T follicular helper (TFH) and T follicular regulatory (TFR) cells have been previously reported, direct models demonstrating the impact of multiple E protein members have been lacking. To suppress all E proteins including E2A, HEB and E2-2, we overexpressed Id1 in CD4 cells using a CD4-Id1 mouse model, to observe any changes in TFH and TFR cell differentiation. Our objective was to gain better understanding of the roles that E proteins and Id molecules play in the differentiation of TFH and TFR cells. The CD4-Id1 transgenic (TG) mice that we constructed overexpressed Id1 in CD4 cells, inhibiting E protein function. Our results showed an increase in the proportion and absolute numbers of Treg, TFH and TFR cells in the spleen of TG mice. Additionally, the expression of surface characterisation molecules PD-1 and ICOS was significantly upregulated in TFH and TFR cells. The study also revealed a downregulation of the marginal zone B cell precursor and an increase in the activation and secretion of IgG1 in spleen B cells. Furthermore, the peripheral TFH cells of TG mice enhanced the function of assisting B cells. RNA sequencing results indicated that a variety of TFH-related functional molecules were upregulated in TFH cells of Id1 TG mice. In conclusion, E proteins play a crucial role in regulating TFH/TFR cell differentiation and function and suppressing E protein activity promotes germinal centre humoral immunity, which has important implications for immune regulation and treating related diseases.
Assuntos
Diferenciação Celular , Proteína 1 Inibidora de Diferenciação , Camundongos Transgênicos , Células T Auxiliares Foliculares , Linfócitos T Reguladores , Animais , Proteína 1 Inibidora de Diferenciação/metabolismo , Proteína 1 Inibidora de Diferenciação/genética , Camundongos , Células T Auxiliares Foliculares/imunologia , Células T Auxiliares Foliculares/metabolismo , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Receptor de Morte Celular Programada 1/metabolismo , Receptor de Morte Celular Programada 1/genética , Proteína Coestimuladora de Linfócitos T Induzíveis/metabolismo , Proteína Coestimuladora de Linfócitos T Induzíveis/genética , Regulação para Cima , Linfócitos B/imunologia , Linfócitos B/metabolismo , Centro Germinativo/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Auxiliares-Indutores/metabolismo , Ativação Linfocitária , Camundongos Endogâmicos C57BL , Imunoglobulina G/imunologiaRESUMO
Drought stress is a predominant abiotic factor leading to decreased alfalfa yield. Genomic ploidy differences contribute to varying adaptation mechanisms of different alfalfa cultivars to drought conditions. This study employed a multi-omics approach to characterize the molecular basis of drought tolerance in a tetraploid variant of alfalfa (Medicago sativa, Xinjiang-Daye). Under drought treatment, a total of 4446 genes, 859 proteins, and 524 metabolites showed significant differences in abundance. Integrative analysis of the multi-omics data revealed that regulatory modules involved in flavonoid biosynthesis, plant hormone signalling transduction, linoleic acid metabolism, and amino acid biosynthesis play crucial roles in alfalfa adaptation to drought stress. The severity of drought led to the substantial accumulation of flavonoids, plant hormones, free fatty acids, amino acids, and their derivatives in the leaves. Genes such as PAL, 4CL, CHI, CHS, PP2C, ARF_3, and AHP_4 play pivotal regulatory roles in flavonoid biosynthesis and hormone signalling pathways. Differential expression of the LOX gene emerged as a key factor in the elevated levels of free fatty acids. Upregulation of P5CS_1 and GOT1/2 contributed significantly to the accumulation of Pro and Phe contents. ERF19 emerged as a principal positive regulator governing the synthesis of the aforementioned compounds. Furthermore, observations suggest that Xinjiang-Daye alfalfa may exhibit widespread post-transcriptional regulatory mechanisms in adapting to drought stress. The study findings unveil the critical mechanisms by which Xinjiang-Daye alfalfa adapts to drought stress, offering novel insights for the improvement of alfalfa germplasm resources.
Assuntos
Adaptação Fisiológica , Secas , Regulação da Expressão Gênica de Plantas , Medicago sativa , Tetraploidia , Medicago sativa/genética , Medicago sativa/fisiologia , Medicago sativa/metabolismo , Adaptação Fisiológica/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Fisiológico/genética , Flavonoides/metabolismo , Flavonoides/biossíntese , Reguladores de Crescimento de Plantas/metabolismo , MultiômicaRESUMO
BACKGROUND: von Willebrand disease (vWD), caused by mutations in the von Willebrand factor (vWF) coding gene, is a disease characterized by abnormal coagulation activity and a severe tendency for hemorrhage. Therefore, identifying mutations in vWF is important for diagnosing congenital vWD. METHODS: We studied a 23-year-old male vWD patient and his parents. Clotting methods were used to determine activated partial thromboplastin time (aPTT), prothrombin time (PT), fibrinogen (FIB) levels, FVIII activity. Chromogenic substrate method was used to determine vWF antigen and activity. The platelet count was determined. Mutations were searched using whole-exome sequencing and certified by Sanger sequencing. Clinical data, including activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), fibrinogen levels, FX activity, FX antigen levels, and the platelet count were collected. A mixing study was performed to eliminate the presence of coagulation factor inhibitors and lupus anticoagulants. Mutations were screened by using whole-exome sequencing (WES) and were verified by using Sanger sequencing. RESULTS: The proband showed severely decreased vWF antigen, vWF activity, and FVIII activity. RIPA (RISTO-CETIN-induced platelet aggregation) was 0%. Data from WES showed that the proband carried compound heterozygous variants vWF: NM_000552.5 (c.3213C>A p.Cys1071Ter) and vWF: NM_000552.5 (c.6598+2T>C). The proband's mother carried variant vWF: NM_000552.5 (c.3213C>A p.Cys1071Ter) while the proband's father carried variant vWF: NM_000552.5 (c.6598+2T>C). All laboratory test indexes of the proband's parents, including vWF antigen, vWF activity, and FVIII activity, were within the normal ranges. CONCLUSIONS: We identified a compound heterozygosis with two novel mutations in vWF (c.3213C>A, c.6598+2T >C) in a family pedigree, and our results demonstrate that the compound heterozygous mutations probably exacerbate vWD.
Assuntos
Doenças de von Willebrand , Fator de von Willebrand , Masculino , Humanos , Adulto Jovem , Adulto , Fator de von Willebrand/genética , Doenças de von Willebrand/diagnóstico , Doenças de von Willebrand/genética , Linhagem , Mutação , Fibrinogênio , ChinaRESUMO
The direct-acting oral anticoagulant dabigatran etexilate (DE) targets thrombin and is used widely to prevent thromboembolism. A 79-year-old man was admitted to the Emergency Department due to anuria for 2 days. An urgent laboratory examination revealed a serum creatinine concentration of 888 µmol/L. He was diagnosed with acute exacerbation of chronic renal insufficiency. During continuous renal replacement therapy (CRRT), the coagulation test showed a severe reduction in the fibrinogen level as well as a significantly prolonged prothrombin time (PT) and activated partial thromboplastin time (APTT). The patient had been taking DE (110 mg twice daily) for a long time and had not suspended the medication or reduced the dose during the worsening of anuria. Therefore, it should be evaluated before considering plasma replacement therapy for the patient, whether the abnormal coagulation parameters were induced by interference of excessive DE. Tentatively, we used activated charcoal to treat the plasma and then retested the fibrinogen, PT, and APTT. Results showed that the coagulation indices nearly returned to normal. The present case indicated that activated charcoal could adsorb DE in plasma effectively and eliminate its interference with coagulation test results, thereby providing support for clinical diagnosis and treatment.
Assuntos
Carvão Vegetal , Dabigatrana , Overdose de Drogas , Humanos , Masculino , Idoso , Carvão Vegetal/uso terapêutico , Overdose de Drogas/diagnóstico , Coagulação Sanguínea/efeitos dos fármacos , Antitrombinas , Testes de Coagulação Sanguínea , Tempo de Protrombina , Anuria/induzido quimicamente , Tempo de Tromboplastina Parcial , Insuficiência Renal Crônica/terapiaRESUMO
With the development of modern medical standards, autoimmune diseases and their associated successive osteoporosis have received increasing attention in recent years. Patients with autoimmune diseases, due to the characteristics of the disease and the prolonged use of glucocorticoid hormone therapy, may affect the bone formation and bone absorption of the patient, followed by severe successive osteoporosis, thereby increasing the risk of osteoporotic vertebral fractures. Vertebral compression fractures of the spine are common fracture types in patients with osteoporotic fractures. Osteoporosis is a common complication after glucocorticoid therapy in patients with autoimmune diseases. Percutaneous vertebroplasty (PVP) and percutaneous kyphoplasty (PKP) are minimally invasive operation and are commonly used surgical methods for the treatment of osteoporotic vertebral compression fractures. However, due to the operation of spinal puncture during the operation, there are serious surgical risks such as bone cement leakage, spinal epidural hemorrhage, subdural hemorrhage, and subarachnoid hemorrhage in both PVP and PKP. As a result, it is necessary to evaluate the patient' s body before surgery carefully, especially in the case of blood coagulation. This article reports a case of autoimmune disease patient admitted to Peking University People' s Hospital due to lumbar 4 vertebral compression fracture combined with Sjögren' s syndrome. The patient' s preoperative examination showed that the activated partial thromboplastin time (APTT) was significantly prolonged. After completing the APTT extended screening experiment and lupus anticoagulant factor testing, the multi-disciplinary team (MDT) of Peking University People' s Hospital jointly discussed the conclusion that the patient' s test results were caused by an abnormal self-immunity anti-copulant lupus (LAC). Based on the results of the laboratory examination, the patient was considered to be diagnosed with combined antiphospholipid syndrome (APS). For such patients, compared with the patient' s tendency to bleed, we should pay more attention to the risk of high blood clotting in the lower limbs of the patient, pulmonary clots and so on. With timely anti-coagulation treatment, the patient safely passed the peripheral period and was successfully discharged from the hospital. Therefore, for patients with autoimmune diseases with prolonged APTT in the perioperative period, doctors need to carefully identify the actual cause and carry out targeted treatment in order to minimize the risk of surgical and perioperative complications and bring satisfactory treatment results to the patients.
Assuntos
Doenças Autoimunes , Fraturas por Compressão , Cifoplastia , Osteoporose , Fraturas por Osteoporose , Fraturas da Coluna Vertebral , Vertebroplastia , Humanos , Fraturas da Coluna Vertebral/cirurgia , Fraturas da Coluna Vertebral/etiologia , Fraturas por Compressão/cirurgia , Vertebroplastia/efeitos adversos , Vertebroplastia/métodos , Tempo de Tromboplastina Parcial , Glucocorticoides , Tempo de Protrombina , Cifoplastia/efeitos adversos , Cifoplastia/métodos , Osteoporose/complicações , Fraturas por Osteoporose/cirurgia , Fraturas por Osteoporose/etiologia , Cimentos Ósseos , Resultado do Tratamento , Estudos RetrospectivosRESUMO
OBJECTIVE: To investigate the expression of layilin (LAYN) in human circulating monocytes and lymphocytes and its clinical significance in systemic lupus erythematosus (SLE). METHODS: Blood samples were collected from 51 SLE patients and 50 healthy controls. Flow cytometry was used to analyze LAYN in lymphocytes and monocyte subsets. Functionally characterized molecules including human HLA, CD74 and CD62L were studied in LAYN+ monocytes. A correlation analysis was conducted between LAYN-related subsets and clinical indicators of SLE such as anti-double-stranded DNA and complements levels. ROC curves were used to explore the potential clinical diagnostic value of LAYN in SLE. RESULTS: LAYN was significantly higher in monocytes than in lymphocytes and higher in CD14+CD16+ monocytes than in CD14-CD16+ and CD14+CD16- monocytes. CD74 was upregulated and CD62L was downregulated in LAYN+ monocytes compared with LAYN- monocytes. The absolute number of LAYN+ monocytes was increased in SLE patients, and the median fluorescence intensity of HLA was decreased. LAYN+ monocytes were positively correlated with complement C4, while decreased CD62L+ percentages in LAYN+ monocytes were negatively correlated with C4. The ROC analysis revealed that the area under the curve (AUCs) for CD62L+ percentages in LAYN+ monocytes, LAYN+ lymphocyte numbers, and LAYN+ monocyte numbers to distinguish SLE from healthy individuals were 0.6245, 0.6196 and 0.6173, respectively. CONCLUSION: LAYN is differentially expressed in monocytes and their subpopulations and has corresponding functional differences. Changes in LAYN expression on monocytes are associated with complement C4 levels in SLE patients. These suggest that LAYN may be involved in the pathogenesis of SLE. ABBREVIATION: ANOVA: analysis of variance; anti-dsDNA: anti-double-stranded DNA; anti-ENA: anti-extractable nuclear antigen; anti-SSA: anti-Sjogren syndrome A; anti-SSB: anti-Sjogren syndrome B; anti-U1RNP: anti-U1 ribonucleoprotein; AUC: area under the ROC curve; CBC: complete blood count; CD62L: L-selectin; CD74/Ii: MHC class II invariant chain; CD44/HCAM: homing cell adhesion molecule; cMos: classical monocytes; CRP: C-reactive protein; CXCR2: C-X-C motif chemokine receptor 2; CXCR4: C-X-C motif chemokine receptor 4; ESR: erythrocyte sedimentation rate; HCs: healthy controls; HA: hyaluronan; HLA: human leukocyte antigen; Ig: immunoglobulin; iMos: intermediate monocytes; LAYN: layilin; MFI: median fluorescence intensity; MIF: migration inhibitory factor; ncMos: nonclassical monocytes; PBMCs: peripheral blood mononuclear cells; ROC: receiver operating characteristic curve; SLE: systemic lupus erythematosus; SLEDAI, SLE disease activity index; Treg: regulatory T cells; WBCs: white blood cells.
Assuntos
Lúpus Eritematoso Sistêmico , Monócitos , Humanos , Leucócitos Mononucleares , Complemento C4 , Anticorpos Antinucleares , Receptores de Quimiocinas , Lectinas Tipo CRESUMO
BACKGROUND: The goal was to clarify the changes of TEG parameters in patients with uterine fibroids and endometrial cancer and the clinical diagnostic values of TEG parameters. METHODS: A total of 57 patients with uterine fibroids and 43 patients with endometrial cancer were included, and their TEG parameters were analyzed and compared with 45 healthy women. Routine coagulation indicators were also collected and compared. For significantly changed TEG indicators, the ROC curves were used to evaluate their diagnostic efficacy and determine the cutoff values. The TEG indicators of patients with endometrial cancer of stag I and II were also compared. RESULTS: APTT, and PT levels in endometrial cancer patients were significantly shorter than those in healthy controls. FIB level in endometrial cancer patients were significantly higher than those in healthy controls. Angle, MA, CI, E, G, and TPI levels were significantly upregulated in endometrial cancer patients while TMA was significantly decreased. According to ROC curve analysis, G and E had a good auxiliary diagnostic efficiency for the detection of uterine fibroids (cutoff value 6,691 d/sec and 133.8 d/sec) and TPI has good sensitivity and specificity for the diagnosis of endometrial cancer (cutoff value 51.3 dyn/cm2). The TEG index of patients with stage I and II endometrial cancer did not reach statistical difference. CONCLUSIONS: Thromboelastography parameters change significantly in patients with endometrial cancer and uterine fibroids.
Assuntos
Neoplasias do Endométrio , Leiomioma , Humanos , Feminino , Tromboelastografia , Testes de Coagulação Sanguínea , Coagulação SanguíneaRESUMO
OBJECTIVE: This study aimed to clarify the expression of HLA-DQ and granulysin in peripheral blood T-cell subsets in patients with chronic hepatitis B virus (CHB) and to evaluate their significance in assisting CHB diagnosis and immune status assessment. METHODS: Peripheral blood from 34 CHB patients, 36 inactive HBsAg carriers and 33 healthy controls were collected, and HLA-DQ and granulysin in a series of T-cell subsets were analysed by flow cytometry. The ability to secrete IL-10 and IFN-γ and the functional T-cell subsets were measured in Treg and CD4 cells expressing HLA-DQ or not. Correlation analyses were further conducted between HLA-DQ/granulysin-related subsets and clinical indicators of HBV infection, and ROC curves were built to evaluate diagnosis efficiency of HLA-DQ-related subsets. RESULTS: HLA-DQ+ percentages in circulating CD4 T cells were downregulated in CHB patients. The proportions of HLA-DQ + Tfh in CHB were upregulated while HLA-DQ+ percentages in Treg were decreased. In terms of function, the IFN-γ secretion ability of CD4 + T cells and IL-10 secretion in Tregs were stronger in HLA-DQ+ than HLA-DQ- subsets. HLA-DQ + CD4 + T cells and HLA-DQ + Treg were negatively correlated with HBV-DNA, while HLA-DQ + Tfh and Tfc cells were positively correlated with HBV-DNA and ALT. HLA-DQ + Treg/Tfh/Tfc could help to distinguish CHB from inactive HBsAg carriers. CONCLUSION: HLA-DQ on T cells can characterize the function of T-cell subsets and analysis of HLA-DQ can help to evaluate immune status and assist in diagnosis of CHB.
Assuntos
Hepatite B Crônica , DNA Viral , Antígenos HLA-DQ , Antígenos de Superfície da Hepatite B , Vírus da Hepatite B , Humanos , Interleucina-10 , Subpopulações de Linfócitos T , Linfócitos T ReguladoresRESUMO
OBJECTIVE: This study aims to elucidate the changes in the percentage of GPR56 and/or granzyme B (GZMB) positive cells in rheumatoid arthritis (RA) CD4 and CD8 T lymphocytes, and to explore their clinical value in diagnosing and reflecting the progression of RA. METHODS: The percentages of GPR56 and/or GZMB positive cells were analyzed in peripheral blood (PB) and spleen T cells in a collagen-induced arthritis (CIA) model established in DBA/1 mice. The percentages of GPR56+ and/or GZMB+ cells were further analyzed in PBs from RA patients and healthy controls. Correlation analysis was performed between clinical indicators and GPR56+, GZMB+, and GPR56+ GZMB+ T cells. Receiver operating characteristic (ROC) curves were used to evaluate the value of GPR56 and GZMB in differentiating active and stable remitting RA. RESULTS: GPR56+ levels were increased in CD4 and CD8 T cells in the PB of CIA mice. The percentages of GPR56+ and GZMB+ cells were increased in both CD4 and CD8 T cell subsets in patients with active RA. GPR56+, GZMB+, and GPR56+ GZMB+ cells were positively correlated with rheumatoid factor and DAS28. ROC analysis revealed that AUCs for GPR56+, GZMB+, and GPR56+ GZMB+ cell percentages to distinguish active RA from stable remission RA were 0.7106, 0.6941, 0.7024, with cut-off values of 16.35, 16.40, 14.80 in CD4 + T cells, and 0.8031, 0.8086, 0.8196 with cut-off values 60.25, 62.15, 40.15 in CD8 + T cells, respectively. CONCLUSIONS: GPR56+ and/or GZMB+ T cells are up-regulated in patients with active RA and reflect their condition. The detection of GPR56 and GZMB is helpful for RA disease assessment.
Assuntos
Artrite Reumatoide , Linfócitos T Citotóxicos , Animais , Artrite Reumatoide/diagnóstico , Artrite Reumatoide/imunologia , Linfócitos T CD4-Positivos , Progressão da Doença , Citometria de Fluxo , Humanos , Camundongos , Camundongos Endogâmicos DBA , Receptores Acoplados a Proteínas GRESUMO
OBJECTIVE: The aim of this study was to review the role of activated carbon (AC) in eliminating the interference of rivaroxaban in the detection of lupus anticoagulants (LAs). METHODS: Normal pooled plasma was obtained as group N1, group N2 took 1 mL plasma from N1 and added AC, group N3 was prepared by mixing normal plasma with rivaroxaban, and group N3 was treated with AC according to our procedure, as group N4. Plasma from 22 patients was collected before and 6-12 h after rivaroxaban therapy, described as group P1 and group P2, respectively, and 1 mL plasma was taken from group P2 and treated with AC, as group P3. Anti-Xa and diluted Russell's viper venom time (dRVVT)/silica clotting time (SCT) index in each group were measured and compared. RESULTS: Rivaroxaban concentrations and anti-Xa had high intercorrelations in group N3, and the levels of anti-Xa and dRVVT/SCT index had high intercorrelations. After treatment with AC, influence of rivaroxaban was removed, with LA and coagulation factor assays not influenced. Rivaroxaban administration could affect LA assay results in patients, with all LA results increased. After treatment with AC, results of anti-Xa and LA tests recovered to the level before rivaroxaban therapy. CONCLUSIONS: We proposed a reference procedure for the LA detection of patients using rivaroxaban by AC, and activated carbon was proven to be a simple product to eliminate the interference of rivaroxaban.
Assuntos
Inibidor de Coagulação do Lúpus , Rivaroxabana , Testes de Coagulação Sanguínea/métodos , Carvão Vegetal , Reações Falso-Positivas , Humanos , Tempo de Tromboplastina Parcial , Tempo de Protrombina , Rivaroxabana/uso terapêuticoRESUMO
The direct-acting oral anticoagulant dabigatran etexilate (DE prevent systemic thromboembolism and cerebral apoplexy in patients with nonvalvular atrial fibrillation. On September 22, 2021, an 86-year-old female patient in Peking University People's Hospital presented with a severely reduced fibrinogen level and significantly prolonged prothrombin time (PT) and activated partial thromboplastin time (APTT). She had a history of hypertension, hyperlipidemia, arrhythmia, and a 1-year history of taking DE (110 mg, p.o., b.i.d.). She had no apparent bleeding tendency but had abnormalities in coagulation markers. She denied taking a DE overdose. Before deciding whether to transfuse plasma for replacement therapy, the attending physician wanted to ascertain if the abnormal results resulted from the drug or the abnormal coagulation mechanism of the patient. Tentatively, we used activated charcoal to treat plasma and then retested her coagulation markers: all coagulation tests nearly returned to normal levels. Hence, the cause was a DE overdose. Re-investigation revealed that she had lived alone in the previous week and possibly mistook the number of doses taken due to confusion. DE was withdrawn, and diuretics (Furosemide injection, 40 mg, QD2) were administered simultaneously to accelerate drug excretion. Five days after drug withdrawal, the coagulation tests returned to normal levels. This case report shows that activated charcoal could be used to show that the coagulation disorder was caused by a DE overdose, thereby providing support for the clinical diagnosis and treatment.
Assuntos
Carvão Vegetal , Dabigatrana , Idoso de 80 Anos ou mais , Coagulação Sanguínea , Testes de Coagulação Sanguínea/métodos , Carvão Vegetal/uso terapêutico , Feminino , Humanos , Tempo de Tromboplastina ParcialRESUMO
BACKGROUND: Mutations in the F10-coding gene can cause factor X (FX) deficiency, leading to abnormal coagulation activity and severe tendency for hemorrhage. Therefore, identifying mutations in F10 is important for diagnosing congenital FX deficiency. METHODS: We studied a 63-year-old male patient with FX deficiency and 10 of his family members. Clotting and immunological methods were used to determine activated partial thromboplastin time (aPTT), prothrombin time (PT), thrombin time (TT), fibrinogen levels, FX activity, and FX antigen levels. The platelet count was determined. A mixing study was performed to eliminate the presence of coagulation factor inhibitors and lupus anticoagulant. Mutations were searched using whole-exome sequencing and certified by Sanger sequencing. RESULTS: Genetic analysis of the proband identified two single-base substitutions: c.1085G>A (p.Ser362Asn) and c.1152C>A (p.Tyr384Ter, termination codon, caused by the DNA sequence TAA). His FX activity and antigen levels were 1.7% and 408.53 pg/mL, respectively; aPTT and PT were 52.3 and 48.0 s, respectively. One brother had the same compound heterozygous mutations, and his FX activity and antigen levels were 1.3% and 465.47 pg/mL, respectively; his aPTT and PT were 65.2 and 54.5 s, respectively. His mother, another brother, and one sister were heterozygous for c.1085G>A (p.Ser362Asn), and his daughter and grandson (6 years old) were heterozygous for c.1152C>A (p.Tyr384Ter). CONCLUSION: The heterozygous variants p.Ser362Asn or p.Tyr384Ter indicate mild FX deficiency, but the compound heterozygous mutation of the two causes severe congenital FX deficiency and bleeding. Genetic analysis of these two mutations may help characterize the bleeding tendency and confirm congenital FX deficiency.
Assuntos
Povo Asiático/genética , Deficiência do Fator X/patologia , Fator X/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Sequência de Bases , Criança , China , Deficiência do Fator X/genética , Feminino , Heterozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Tempo de Tromboplastina Parcial , Linhagem , Polimorfismo de Nucleotídeo Único , Tempo de ProtrombinaRESUMO
E proteins, a subset of basic helix-loop-helix (bHLH) proteins, are transcription activators and their functions are inhibited by DNA-binding inhibitor (Id) 1-4. Studies have shown that Treg levels are decreased in Id3 knockout mice. Mice over-expressing Id1 in CD4 T cells possessed a greater number of regulatory T cells (Treg) and exhibited attenuated experimental autoimmune encephalomyelitis (EAE). The significance of Id proteins in human systemic lupus erythematosus (SLE) remains unclear. In this study, we systematically analyzed Id transcription in naïve, memory CD4 cells and regulatory T cells in peripheral blood mononuclear cells (PBMCs) in patients with active or inactive SLE. In parallel, Treg subsets in PBMCs were analyzed using different strategies. Id expression levels were correlated with Treg numbers as well as clinical indicators. We found that Id genes expressed in human peripheral CD4 cells were mainly Id2 and Id3. Id3 levels were significantly elevated in CD4+CD25hi T cells of patients with active SLE. Likewise, Id3 levels were positively correlated with increased CD4+FoxP3+ and CD4+Helios+FoxP3+ Treg cells in these patients. Id3 levels were found to be positively correlated with erythrocyte sedimentation rate (ESR), lupus anticoagulant (LAC), ribosomal antibody and SLE Disease Activity Index (SLEDAI) in patients with active SLE. Mice overexpressing Id1 in CD4+ T cells possessed significantly higher Treg levels in spleen and lower autoantibody concentrations in serum. Our results suggest that during the pathogenesis of SLE, up-regulation of Id3 can promote Treg differentiation to play an inhibitory effect on autoimmune responses.
Assuntos
Proteínas Inibidoras de Diferenciação/metabolismo , Lúpus Eritematoso Sistêmico/diagnóstico , Proteínas de Neoplasias/metabolismo , Subpopulações de Linfócitos T/imunologia , Linfócitos T Reguladores/imunologia , Animais , Autoanticorpos/sangue , Autoanticorpos/imunologia , Estudos de Casos e Controles , Modelos Animais de Doenças , Voluntários Saudáveis , Humanos , Proteína 1 Inibidora de Diferenciação/genética , Proteína 1 Inibidora de Diferenciação/metabolismo , Proteínas Inibidoras de Diferenciação/análise , Proteínas Inibidoras de Diferenciação/genética , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/induzido quimicamente , Lúpus Eritematoso Sistêmico/imunologia , Camundongos Knockout , Camundongos Transgênicos , Proteínas de Neoplasias/análise , Índice de Gravidade de Doença , Subpopulações de Linfócitos T/metabolismo , Linfócitos T Reguladores/metabolismo , Terpenos/administração & dosagem , Terpenos/imunologia , Regulação para Cima/imunologiaRESUMO
AIM: This study was designed to investigate the levels of circulating regulatory T cells (Tregs), and their functional subpopulations and related cytokines in chronic hepatitis B patients (CHB) and inactive hepatitis B surface antigen carriers. METHODS: The peripheral blood of 24 hepatitis B virus inactive carriers, 26 CHB patients, and 34 healthy controls was collected and analyzed by flow cytometry for Tregs and CD4+ CXCR5+ FoxP3+ follicular regulatory T cells. Interleukin (IL)-10, transforming growth factor-ß, and IL-21 levels in plasma were determined by enzyme-linked immunosorbent assay. Proportions of functional Treg subpopulations were analyzed by staining of Helios, CD45RA and FoxP3, TIGIT, and CD226, and the correlations between Treg subsets and clinical indicators were explored. RESULTS: CD4+ FoxP3+ levels in the peripheral blood of CHB patients were significantly increased, and the inhibitory ability of Tregs in CHB patients for cytokine secretion was stronger, and CD4+ CXCR5+ FoxP3+ follicular Tregs were also significantly higher than inactive carriers and healthy controls. Transforming growth factor-ß and IL-10 in the plasma of CHB patients were significantly higher than those of healthy controls, with IL-21 levels not significantly changed. Circulating CD4+ CXCR5-FoxP3+ Treg cells in CHB patients were positively correlated with hepatitis B surface antigen, hepatitis B e antigen, and hepatitis B virus DNA. The proportions of Helios+ FoxP3+ , CD45RA- FoxP3hi , and CD226- TIGIT+ functional subpopulations in CD4+ CXCR5- FoxP3+ Tregs in CHB patients were significantly increased, and they were significantly correlated with clinical indicators. CONCLUSIONS: Circulating Tregs in CHB patients not only have elevated levels, but their follicular Treg subpopulations are also increased, and Tregs tend to have stronger immunosuppressive functions.
RESUMO
Hemocoagulase is often used for hemostasis in patients with bleeding and hemorrhagic diseases, and to avoid or stanch bleeding after surgery. Herein, three patients with hepatic diseases suffering from hypofibrinogenemia were treated with hemocoagulase agkistrodon (HCA) in Peking University People's Hospital during September 2018. All the 3 patients were chronic hepatitis B patients: Patient 1 presented with hepatic carcinoma and chronic hepatitis B, and right hepatectomy was performed; patient 2 presented with chronic hepatitis B and gastrointestinal bleeding; patient 3 presented with chronic hepatitis B, acute liver failure with hematemesis, and was awaiting liver transplantation. All three patients were percutaneously injected with HCA to prevent late-onset bleeding. After HCA was discontinued, coagulation was restored to > 60 mg/dL on day 6, without injection of fibrinogen. HCA significantly reduced the need for fibrinogen in patients with hepatic diseases, and the level of fibrinogen should be carefully monitored in clinical applications.
Assuntos
Afibrinogenemia , Agkistrodon , Batroxobina/uso terapêutico , Hemostáticos/uso terapêutico , Hepatopatias/tratamento farmacológico , Animais , Fibrinogênio , HumanosRESUMO
The plateau zokor (Eospalax baileyi) is a species of subterranean rodent endemic to the Tibetan Plateau. It is well adapted to the cold and hypoxic and hypercapnic burrow. To study the oxygenation properties of plateau zokor hemoglobins (Hbs), we measured intrinsic Hb-O2 affinities and their sensitivities to pH (Bohr effect); CO2; Cl-, 2,3-diphosphoglycerate (DPG); and temperature using purified Hbs from zokor and mouse. The optimal deoxyHb model of plateau zokor was constructed and used to study its structural characteristics by molecular dynamics simulations. O2 binding results revealed that plateau zokor Hbs exhibit remarkably high intrinsic Hb-O2 affinity, low CO2 effects compared with human and the relatively low anion allosteric effector sensitivities (DPG and Cl-) at normal temperature, which would safeguard the pulmonary Hb-O2 loading under hypoxic and hypercapnic conditions. Furthermore, the high anion allosteric effector sensitivities at low temperature and low temperature sensitivities of plateau zokor Hbs would facilitate the releasing of O2 in cold extremities and metabolic tissues. However, the high Hb-O2 affinity of plateau zokor is not compensated by high pH sensitivity as the Bohr factors of plateau zokor Hbs were as low as those of mouse. The results of molecular dynamics simulations revealed the reduced hydrogen bonding between the α1ß1- and α2ß2-dimer interface of deoxyHb in zokor compared with mouse. It may be the primary mechanism of the high intrinsic Hb-O2 affinities in zokor. Specifically, substitution of the 131SerâAsn in the α2-chain weakened the connection between α1- and ß2-subunit.
Assuntos
Hemoglobinas/metabolismo , Consumo de Oxigênio , Oxigênio/sangue , Oxiemoglobinas/metabolismo , Animais , Biomarcadores/sangue , Temperatura Corporal , Dióxido de Carbono/sangue , Hemoglobinas/química , Humanos , Ligação de Hidrogênio , Concentração de Íons de Hidrogênio , Masculino , Camundongos , Simulação de Dinâmica Molecular , Oxiemoglobinas/química , Ligação Proteica , Estrutura Terciária de Proteína , Subunidades Proteicas , Roedores , Especificidade da Espécie , Relação Estrutura-AtividadeRESUMO
Objective: This study aims to clarify the changes and clinical significance of red cell distribution width (RDW) during HBV-related chronic diseases, including inactive hepatitis B virus (HBV) carriers, HBV immune tolerant individuals, chronic hepatitis B (CHB) patients and HBV-related hepatocirrhosis patients. Methods: RDW was measured 288 CHB patients, 100 patients with hepatitis B e antigen(HBeAg)-negative chronic HBV infection (inactive carriers), 92 patients with HBeAg-positive chronic HBV infection (immune tolerant), and 272 patients with HBV-related hepatocirrhosis. Their RDW changes were compared with 160 healthy controls. Correlations between RDW and clinical indicators were conducted. For HBeAg+ CHB patients, RDW was measured before and after antiviral therapy. The efficiency of RDW to distinguish hepatocirrhosis from CHB and/or inactive carriers was evaluated by receiver operating characteristic (ROC) curves. Results: RDW was higher in hepatocirrhosis patients than other groups of patients and healthy controls. Besides, HBeAg+ CHB patients possessed higher RDW than HBeAg- CHB patients. For HBeAg+ patients that underwent HBeAg seroconversion after antiviral therapy, RDW was decreased. RDW was positively correlated with total bilirubin and Child-Pugh scores and negatively correlated with albumin among hepatocirrhosis patients. The areas under the curve (AUC) of ROC curves to distinguish hepatocirrhosis from CHB patients was 0.7040 for RDW-standard deviation (RDW-SD) and 0.6650 for RDW-coefficient of variation (RDW-CV), and AUC to distinguish hepatocirrhosis from inactive carriers was 0.7805 for RDW-SD and 0.7991 for RDW-CV. Conclusions: RDW is significantly increased in HBeAg+ CHB patients and patients with HBV-related hepatocirrhosis and could reflect their severity. RDW could help to distinguish hepatocirrhosis from CHB patients and inactive HBV carriers.
Assuntos
Antígenos E da Hepatite B/sangue , Vírus da Hepatite B/patogenicidade , Hepatite B Crônica/sangue , Cirrose Hepática/sangue , Adulto , Diagnóstico Diferencial , Índices de Eritrócitos/imunologia , Feminino , Antígenos E da Hepatite B/imunologia , Vírus da Hepatite B/imunologia , Vírus da Hepatite B/metabolismo , Hepatite B Crônica/imunologia , Hepatite B Crônica/patologia , Hepatite B Crônica/virologia , Humanos , Cirrose Hepática/imunologia , Cirrose Hepática/patologia , Cirrose Hepática/virologia , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Urine sediment parameters of pregnant women are different from those of non-pregnant women, and it is necessary to establish reference intervals for pregnant women. The aim of this study was to establish reference intervals of white blood cell (WBC), red blood cell (RBC), bacteria (BACT), squamous epithelial cell (EC), small round epithelial cell (SRC), and mucous strands (MUS) for urine sediment test of pregnant women using a UF-1000i analyzer as the detection device. The differences between pregnant women and non-pregnant women in terms of the aforementioned parameters as well as the differences of such parameters in different trimesters of pregnancy were clarified. METHODS: The experimental subjects were divided into two groups: the experiment group (612 healthy pregnant women) and the control group (582 healthy non-pregnant women). Subjects of both groups are women between the age of 22 and 46. The urine specimens were analyzed using the Sysmex UF-1000i analyzer, followed by manual correction. A statistical analysis was performed by SPSS 22.0. Results were considered significant at p < 0.01. RESULTS: The pregnancy reference intervals of WBC, RBC, BACT, EC, SRC, and MUS were 0 ~ 30/µL, 0 ~ 23/µL, 0 ~ 698/µL, 0 ~ 28/µL, 0 ~ 8/µL, and 0 ~ 3/µL, respectively. In the experiment group, the concentrations of WBC, BACT, EC, and SRC were significantly higher than those of the control group (p < 0.01), while the concentrations of RBC and MUS were significantly lower than those of the control group (p < 0.01). The inter-trimester differences in terms of the concentrations of WBC, BACT, EC, and SRC were statistically indistinguishable (p > 0.05). However, the concentration of RBC was significantly lower with the increase of trimester of pregnancy (the comparison between the first trimester with the second trimester: p = 0.000 < 0.01; the comparison between the second trimester and the third trimester: p = 0.004 < 0.01). The WBC, BACT, EC, and SRC had moderate intercorrelations (0.569 ~ 0.681, p < 0.01). CONCLUSIONS: There were significant differences in the aforementioned parameters between the two groups. The intervals of WBC, RBC, BACT, EC, SRC, and MUS for urine sediment analysis of healthy pregnant women using a UF-1000i should be established.
Assuntos
Primeiro Trimestre da Gravidez/urina , Segundo Trimestre da Gravidez/urina , Trimestres da Gravidez/urina , Urinálise/instrumentação , Urinálise/métodos , Adulto , Contagem de Eritrócitos , Feminino , Humanos , Contagem de Leucócitos , Pessoa de Meia-Idade , Gravidez , Valores de Referência , Reprodutibilidade dos Testes , Urina/microbiologia , Adulto JovemRESUMO
Intracranial teratoma, a subtype of non-germinomatous germ cell tumors, is rare in adults. Clinical presentation of intracranial teratomas varies according to where they grow. In particular, cases of spontaneous ruptures of intracranial teratoma are sporadic. This study reports the case of an adult with a spontaneously ruptured mature teratoma in the cerebellar vermis, which was comorbid with a dermal sinus tract and subcutaneous lipoma. Before surgery, because the images were atypical of a teratoma, the patient was misdiagnosed as having vascular malformation rupture and bleeding in the cerebellar vermis. Due to the patient's level of consciousness dropping drastically to a coma, a craniotomy was performed. During the surgery, the tumor was observed to be a mixed cystic and solid mass. The liquid in the cyst was dark green and with a fatty component. The solid part had a tough texture and comprised hair, fat, cartilage, and calcification components. Post-surgery multipoint biopsy proved that it was a mature teratoma and that it was connected to a subcutaneous lipoma through the dermal sinus tract across the occipital bone. After proactive treatment, the patient's prognosis was favorable.