ABSTRACT
We studied the mechanisms by which microRNA-126 regulates proliferation and migration of human umbilical vein endothelial cells (HUVEC) cultured in a medium with high glucose concentration and treated with chemokine CXCL8. Cell proliferation, apoptosis, and migration were analyzed by the CCK-8 assay, Annexin V-PI staining, and Transwell assay, respectively. The ratios of p-ERK/ERK, p-P38/P38, p-JNK/JNK were determined by ELISA. HUVEC cells cultured in the presence of high glucose concentration (30 mmol/ml) and treated with CXCL8 (50 ng/ml) demonstrated more intensive proliferation, migration, and p-ERK/ERK, p-P38/P38, and p-JNK/JNK ratios and significantly lower apoptosis rate than control cells (high glucose, no treatment) and cells treated with CXCL8 and transfected with microRNA-126-mimic. Thus, microRNA-126 regulates proliferation and migration of HUVEC cells cultured in the presence of high glucose concentrations and treated with CXCL8 through inhibition of MAPK signaling pathway.
Subject(s)
Glucose/pharmacology , Human Umbilical Vein Endothelial Cells/drug effects , Interleukin-8/pharmacology , MicroRNAs/physiology , Apoptosis/drug effects , Apoptosis/genetics , Cell Movement/drug effects , Cell Movement/genetics , Cell Proliferation/drug effects , Cell Proliferation/genetics , Cells, Cultured , Culture Media/chemistry , Culture Media/pharmacology , Dose-Response Relationship, Drug , Human Umbilical Vein Endothelial Cells/physiology , Humans , MAP Kinase Signaling System/drug effects , MAP Kinase Signaling System/genetics , MicroRNAs/genetics , Signal Transduction/drug effects , Signal Transduction/geneticsABSTRACT
Objective: To explore the phenotypes and genotypes of molybdenum cofactor deficiency type B (MoCD-B) manifested as Leigh-like syndrome. Methods: The clinical data, laboratory tests, neuroimaging and gene results of one patient diagnosed as MoCD-B at Beijing Children's Hospital and Hebei Children's Hospital in December 2018 were collected. Related literature was searched and reviewed at Wanfang Data Knowledge Service Platform, China National Knowledge Infrastructure and PubMed (up to September 2020) by using terms "MOCS2" "molybdenum cofactor deficiency" "Leigh-like syndrome,MOCS2" "molybdenum cofactor deficiency, Leigh-like syndrome". The phenotypes and genotypes of MoCD-B were summarized. Results: A 7 months and 14 days old boy with the chief complaint of "cough for 6 days, abnormal posture for 4 days and fever for 2 days" was admitted to Hebei Children' Hospital on December 2018. His abnormal posture presented as opisthotonos accompanied with dysphagia, without seizures. His previous psychomotor development was described as normal. He was born at term after an uneventful pregnancy to non-consanguineous parents. Blood test showed a slightly increased lactic acid and a significantly decreased uric acid. Urine metabolism test showed an obviously increased xanthine and hypoxanthine. Brain magnetic resonance imaging showed hyperintense signal on T2 weighted image and fluid attenuated inversion recovery in bilateral globus pallidus and pedunculus cerebri. The patient was diagnosed with Leigh-like syndrome. No obvious improvement was achieved after cocktail therapy and symptomatic treatment. The whole exome sequencing showed that the patient carried a homozygous variant of MOCS2 gene, c.19G>T(p.Val7Phe), which was a previously reported pathogenic site in the literature and could cause MoCD-B. His parents carried a heterozygous variant respectively. A total of 41 MoCD-B cases with MOCS2 gene variants were collected through literature review and our study, among which 30 cases had full medical records. The onset ages of 23 (77%) cases were in neonate, manifesting with severe encephalopathy, including neonatal-onset intractable seizures, developmental delay, laboratory abnormalities included very low levels of serum and urinary uric acid, increased urinary levels of xanthine and hypoxanthine. Cranial imaging showed cerebral atrophy, cystic encephalomalacia, etc. The onset ages of 7 patients varied from 5 months to 23 years. Four cases had normal psychomotor development before disease onset. Neurological disorders appeared acutely or exacerbated after external triggers and all of them had basal ganglia involvement. Among the 30 cases, 3 cases had a relatively milder phenotype with the ability of brief communication and walking without or with support. Conclusions: Molybdenum cofactor deficiency is a rare disease. Most cases had severe phenotypes and poor outcomes, but some cases may have mild phenotype. MoCD-B caused by MOCS2 gene variants may manifest as Leigh-like syndrome with a normal psychomotor development before the trigger of infection strike. Hypouricemia, xanthinuria and hypoxanthinuria can be indicators of the disease. The presence of MOCS2 gene variants would confirm a final diagnosis.
Subject(s)
Metal Metabolism, Inborn Errors , Child , Child, Preschool , China , Homozygote , Humans , Infant , Infant, Newborn , Male , Metal Metabolism, Inborn Errors/diagnosis , Metal Metabolism, Inborn Errors/genetics , PhenotypeABSTRACT
Maxillary sinus augmentation is an effective procedure to gain bone height for implant placement in an atrophic posterior maxilla. But maxillary sinus diseases are prevalent in patients scheduled for sinus lift procedures. The presence of these diseases may increase the difficulties in performing the surgery and the risk of developing postoperative complications. This paper summarizes and introduces the common maxillary sinus mucosa diseases related to maxillary sinus augmentation.
Subject(s)
Maxillary Sinus , Mucous Membrane , Sinus Floor Augmentation , Dental Implantation, Endosseous , Dental Implants , Humans , Lifting , Maxilla , Maxillary Sinus/pathology , Mucous Membrane/pathology , Sinus Floor Augmentation/adverse effectsABSTRACT
Objective: To establish comprehensive laboratory reference intervals for Chinese children. Methods: This was a cross-sectional multicenter study. From June 2013 to December 2014, eligible healthy children aged from 6-month to 17-year were enrolled from 20 medical centers with informed consent. They were assessed by physical examination, questionnaire survey and abdominal ultrasound for eligibility. Fasting blood samples were collected and delivered to central laboratory. Measurements of 15 clinical laboratory parameters were performed, including estradiol (E2), testosterone(T), luteinizing hormone(LH), follicle-stimulating hormone(FSH), alanine transaminase(ALT), serum creatinine(Scr), cystatin C, immunoglobulin A(IgA), immunoglobulin G(IgG), immunoglobulin M(IgM), complement (C3, C4), alkaline phosphatase(ALP), uric acid(UA) and creatine kinase(CK). Reference intervals were established according to central 95% confidence intervals for reference population, stratified by age and sex. Results: In total, 2 259 children were enrolled. Finally, 1 648 children were eligible for this study, including 830 boys and 818 girls, at a mean age of 7.4 years. Age- and sex- specific reference intervals have been established for the parameters. Reference intervals of sex hormones increased gradually with age. Concentrations of ALT, cystatin C, ALP and CK were higher in children under 2 years old. Serum levels of sex hormones, creatinine, immunoglobin, CK, ALP and urea increased rapidly in adolescence, with significant sex difference. In addition, reference intervals were variable depending on assay methods. Concentrations of ALT detected by reagents with pyridoxal 5'-phosphate(PLP) were higher than those detected by reagents without PLP. Compared with enzymatic method, Jaffe assay always got higher results of serum creatinine, especially in children younger than 9 years old. Conclusion: This study established age- and sex- specific reference intervals, for 15 clinical laboratory parameters based on defined healthy children.
Subject(s)
Blood Chemical Analysis , Reference Values , Adolescent , Child , Child, Preschool , Cross-Sectional Studies , Estradiol/blood , Female , Follicle Stimulating Hormone/blood , Humans , Infant , Luteinizing Hormone/blood , MaleABSTRACT
Objective: To study effects of cyclin D1 overexpression on the proliferation and differentiation of cervical squamous cell carcinoma SiHa cells and to investigate related signaling molecules. Methods: Primers were designed to amplify the full length of cyclin D1 gene and cyclin D1 gene was amplified by PCR for constructing pcDNA3.1 plasmid vector. The construct was then transfected into SiHa cells, and the cells with stable overexpression of cyclin D1 were established, cyclin D1 gene and protein expression were detected by RT-PCR and Western blot, respectively. Cell growth curve was documented by MTT assay. CK7, E-cadherin, vimentin, Snail gene and protein expression in transfected cells were detected by RT-PCR and Western blot. RT-PCR was used to detect the mRNA expression of proliferation and differentiation-related genes like CDK4, CDK2, p21, p27, cyclin E, Rb, E2F, E6/E7 and Ki-67. After synchronization of cells, RT-PCR was used to detect of cyclin D1 and p21 mRNA expression at different time points of the cell cycle. Results: The G-3 cells with cyclin D1 overexpression were successfully established. The growth curve and Ki-67 mRNA expression accelerated in G-3 cells.Vimentin and Snail expression significantly increased at both gene and protein levels, while E-cadherin, CK7 gene and protein expression significantly decreased, indicating epithelial mesenchymal transitionoccurred in G-3 cells.Meanwhile, mRNA expression of cyclin D1, CDK4, CDK2, p21, p27, cyclin E, E2F and Rb increased, while E6/E7 and p16 showed no significant change. The expression trends of p21 and cyclin D1 were almost identical with fluctuation at different time points in the cell cycle. Conclusions: Overexpression of cyclin D1 induced by gene transfection promotes proliferation and epithelial mesenchymal transition in SiHa cells.The process is accompanied by up-regulation of CDK4, CDK2, p21, p27 and cyclin E genes.p21 expression increases synchronously with cyclin D1, suggesting a regulatory role in epithelial mesenchymal transition by affecting expression of vimentin in G-3 cells.
Subject(s)
Carcinoma, Squamous Cell/genetics , Cell Cycle Proteins/genetics , Cyclin D1/genetics , Epithelial-Mesenchymal Transition , Gene Expression Regulation, Neoplastic/genetics , Uterine Cervical Dysplasia/genetics , Uterine Cervical Neoplasms/genetics , Amplified Fragment Length Polymorphism Analysis , Antigens, CD , Cadherins , Carcinoma, Squamous Cell/metabolism , Cell Proliferation , Cyclin D1/metabolism , Cyclin E , Cyclin-Dependent Kinase 2 , Female , Humans , Oncogene Proteins , Up-Regulation , Uterine Cervical Neoplasms/metabolism , Uterine Cervical Dysplasia/metabolismABSTRACT
Objective: To investigate the clinically and genetic characteristics of children with Leigh syndrome. Method: Patients with clinically diagnosed Leigh syndrome(LS)in the department of Neurology, Beijing Children's Hospital from January 2013 to February 2016 underwent the mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) detecting with next generation sequencing (NGS) technology. The clinical data of gene confirmed cases were retrospectively collected and analyzed. The differences in the onset age, clinical manifestations, lactic acid level and MRI results between the mtDNA variation and nDNA variation were compared and analyzed.t test, Chi-square test and Fisher's exact test were used for statistical analysis. Result: Thirty-five cases were diagnosed by gene detection, including 20 males and 15 females. The median onset age was 1 year (ranging from the neonatal period to 4.4 years old). The age of onset within 2 years accounted for 74%(26 cases). The onset age of initial symptoms, including developmental delay, developmental regression, and seizures, were 6 (4, 12) months, 12 (8, 14) months, and 6 (1, 23) months respectively. The onset age of ptosis, extrapyramidal symptoms and ataxia were 26 (18, 44) months, 28 (23, 40) months and 28 (19, 35) months, respectively. There were significant differences in the onset age between the three groups (H=21.919, P=0.01). Within the 35 cases, 29 were manifested with developmental delay (83%), 26 with dystonia (74%), 18 with growth retardation, 15 with myasthenia, 13 with developmental regression, 11 with dysphagia, 10 with feeding difficulties, 4 with skeletal dysplasia, and 2 with digestive tract symptoms; nystagmus and respiratory abnormalities were observed in 9 cases respectively; extrapyramidal symptoms, peripheral nerve injury, ptosis, seizures were observed in 8 cases respectively; and ataxia, ophthalmoplegia and hypertrichiasis were found in 5 cases respectively.The blood lactic acid was measured in 32 LS patients, within which 23 cases (72%) had increased results; 8 out of 11 cases who underwent were cerebrospinal fluid lactic acid test had increased results. The results of neuroimaging revealed that all the patients were involved in the brainstem and (or) basal ganglia, of whom 27 (77%) had brainstem involvement, 24 (69%) had basal ganglia involvement. Thirteen out of 14 patients who had medulla oblongata involvement had nDNA variation; while 7 out of 8 patients with cerebellar involvement had nDNA variation. Genetic etiology was confirmed in all patients, among whom there were 17 cases (49%) with mtDNA mutation, including 8993T>C/G (n=5), 14487T>C (n=4), 13513G>A (n=2), 9176T>C, 10158T>C, 3697G>A, 10191T>C, 14459A>G and 11777C>A (n=1) respectively. Remaining 18 cases(51%) had nDNA mutation, including SURF1 gene(n=10), PDHA1 gene(n=3) and one case each of NDUFV1, NDUFAF6, NDUFAF5, NDUFS1 and COQ7 genes. In this study, 27 types of mutations were founded, 15 of which had not been previously reported. Respiratory chain gene mutations have been found in 31 cases(89%); 3 cases had PDHc gene mutations, and 1 case had other mutation. Conclusion: LS usually occurs in infants. The most common primary symptoms are age-dependent abnormal movements, ocular symptoms, and seizures. Respiratory chain defects is the most common causes of LS.SURF1 is the most common variation, followed by 8993T>C/G, 14487 T>C and 13513G>A mutation.
Subject(s)
Leigh Disease/genetics , Mutation , Age of Onset , Child , Child, Preschool , DNA, Mitochondrial , Dystonia , Female , High-Throughput Nucleotide Sequencing , Humans , Infant , Leigh Disease/diagnosis , Magnetic Resonance Imaging , Male , Nystagmus, Pathologic , Retrospective StudiesABSTRACT
Topological Weyl semimetal (TWS), a new state of quantum matter, has sparked enormous research interest recently. Possessing unique Weyl fermions in the bulk and Fermi arcs on the surface, TWSs offer a rare platform for realizing many exotic physical phenomena. TWSs can be classified into type-I that respect Lorentz symmetry and type-II that do not. Here, we directly visualize the electronic structure of MoTe2, a recently proposed type-II TWS. Using angle-resolved photoemission spectroscopy (ARPES), we unravel the unique surface Fermi arcs, in good agreement with our ab initio calculations that have nontrivial topological nature. Our work not only leads to new understandings of the unusual properties discovered in this family of compounds, but also allows for the further exploration of exotic properties and practical applications of type-II TWSs, as well as the interplay between superconductivity (MoTe2 was discovered to be superconducting recently) and their topological order.
ABSTRACT
To investigate the role of T-helper cells/Treg (Th17/Treg) and morbidity factors related to primary nephritic syndrome (PNS) in children, as well as the influence of ox-low density lipoprotein (ox-LDL) on Th17/Treg expression in children with PNS. To clarify the pathogenesis of PNS in children, 50 children with PNS treated in our hospital were enrolled in the study group. Additionally, 20 healthy children who came to our hospital for physical examination during the same period were enrolled in the control group. Th17 and Treg cells in children belonging to the two groups were detected by flow cytometry; the numbers of Th17/Treg cells in peripheral blood mononuclear cells at different concentrations of ox-LDL were detected simultaneously. Ox-LDL can affect the number of Th17/Treg cells in peripheral blood mononuclear cells, and both cell types decreased with increasing concentration of ox-LDL, with the numbers being significantly lower in the control group. However, the decrease in the number of Th17 cells was statistically insignificant (P > 0.05), whereas the decrease in Treg cells was more obvious and statistically significant (P < 0.05). The effect of ox-LDL the number of Treg cells was stronger than that on Th17 cells. We concluded that the imbalance of Th17/Treg cells influenced by high and low ox-LDL concentrations in children with PNS might be the immunological basis of the disease.
Subject(s)
Lipoproteins, LDL/blood , Nephrotic Syndrome/immunology , T-Lymphocytes, Regulatory/immunology , Th17 Cells/immunology , Cell Count , Child, Preschool , Female , Flow Cytometry , Humans , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/metabolism , Lipoproteins, LDL/immunology , Male , Nephrotic Syndrome/blood , Nephrotic Syndrome/pathology , T-Lymphocytes, Regulatory/metabolism , Th17 Cells/metabolismABSTRACT
OBJECTIVE: To investigate the JNK/AP-1 signaling pathway of airway mucus hypersecretion of severe pneumonia under respiratory virus (RSV) infection. PATIENTS AND METHODS: Total of 56 severe pneumonia children under RSV infection were selected. Reverse transcription polymerase chain reaction (RT-PCR) was performed to measure the expression quantity of MUC5B mRNA and MUC5AC mRNA, and ELISA was used to measure the expression quantity of MUC5AC and MUC5B proteins. Following that, the children were divided into airway mucus hypersecretion group (n = 37) and non-hypersecretion group (n = 19). Western blotting was performed to detect the expression levels of JNK1/2, p-JNK1/2 and AP-1 proteins. RESULTS: Expression of MUC5AC and MUC5B proteins, and MUC5AC mRNA and MUC5B mRNA in the airway mucus hypersecretion group were significantly higher than those in the non-hypersecretion group (p < 0.05). The expression levels of JNK1/2, p-JNK1/2 and AP-1 proteins in airway mucus hypersecretion group were higher than those in the non-hypersecretion group (p < 0.05). CONCLUSIONS: MUC5AC and MUC5B can be used as marker molecules of airway mucus hypersecretion. Airway mucus hypersecretion of severe pneumonia induced by RSV might be related to the activation of JNK/AP-1 signaling pathway.
Subject(s)
MAP Kinase Signaling System , Mucus/metabolism , Pneumonia/genetics , Respiratory System/metabolism , Respirovirus Infections/genetics , Child , Child, Preschool , Female , Humans , Infant , MAP Kinase Signaling System/genetics , Male , Mucin 5AC/genetics , Mucin 5AC/metabolism , Mucin-5B/genetics , Mucin-5B/metabolism , Pneumonia/metabolism , Pneumonia/virology , Respiratory System/virology , Respirovirus Infections/complications , Respirovirus Infections/metabolism , Severity of Illness Index , Transcription Factor AP-1/genetics , Transcription Factor AP-1/metabolismABSTRACT
Diverse non-cardiac drugs adversely influence cardiac electrophysiology by inhibiting repolarising K(+) currents mediated by channels encoded by the human ether-a-go-go-related gene (hERG). In this study, pharmacological blockade of hERG K(+) channel current (I(hERG)) by a novel investigative serotonin-selective reuptake inhibitor (SSRI), CONA-437, was investigated. Whole-cell patch-clamp measurements of I(hERG) were made from human embryonic kidney (HEK 293) cells expressing wild-type (WT) or mutant forms of the hERG channel. With a step-ramp voltage-command, peak I(hERG) was inhibited with an IC(50) of 1.34 µM at 35 ±1°C; the IC(50) with the same protocol was not significantly different at room temperature. Voltage-command waveform selection had only a modest effect on the potency of I(hERG) block: the IC50 with a ventricular action potential command was 0.72 µM. I(hERG) blockade developed rapidly with time following membrane depolarisation and showed a weak dependence on voltage, accompanied by a shift of ≈ -5 mV in voltage-dependence of activation. There was no significant effect of CONA-437 on voltage-dependence of I(hERG) inactivation, though at some voltages an apparent acceleration of the time-course of inactivation was observed. Significantly, mutation of the S6 aromatic amino acid residues Y652 and F656 had only a modest effect on I(hERG) blockade by CONA-437 (a 3-4 fold shift in affinity). CONA-437 at up to 30 µM had no significant effect on either Nav1.5 sodium channels or L-type calcium channels. In conclusion, the novel SSRI CONA-437 is particularly notable as a gating-dependent hERG channel inhibitor for which neither S6 aromatic amino-acid constituent of the canonical drug binding site on the hERG channel appears obligatory for I(hERG) inhibition to occur.
Subject(s)
Dimethylamines/pharmacology , Ether-A-Go-Go Potassium Channels/physiology , Potassium Channel Blockers/pharmacology , Pyridines/pharmacology , Selective Serotonin Reuptake Inhibitors/pharmacology , Animals , CHO Cells , Calcium Channels, L-Type/physiology , Cell Line , Cricetinae , Cricetulus , HEK293 Cells , Humans , NAV1.5 Voltage-Gated Sodium Channel/genetics , NAV1.5 Voltage-Gated Sodium Channel/physiology , RatsABSTRACT
The in vitro culture of lymphatic smooth muscle cells (SMCs) is a crucial step in studying their function and involvement in disease. Yet there is no efficient approach available so far because of the difficulties posed by the small size of most lymphatic vessels. We present a simple yet efficient method for isolating and culturing SMCs of collecting lymphatic vessels from guinea pig mesenteric tissue. In our approach, thin lymphatic vessels were digested twice from adventitia to media to release SMCs, which were then cultured by traditional methods. The lymphatic SMCs we cultured did not exhibit contact inhibition and demonstrated typical SMCs characteristics under light microscope, electron microscope and by immunohistochemical studies. This method is applicable to the culturing of lymphatic SMCs from other organs and provides useful materials for physiological and pathological lymphatic studies.
Subject(s)
Lymphatic Vessels/cytology , Mesentery/cytology , Myocytes, Smooth Muscle/ultrastructure , Actins/metabolism , Adipocytes , Animals , Blood Cells , Cell Separation , Cells, Cultured , Culture Techniques/methods , Endothelial Cells , Female , Fibroblasts , Fluorescent Antibody Technique , Guinea Pigs , Immunohistochemistry , Male , Microscopy, Electron , Models, Animal , Myocytes, Smooth Muscle/metabolism , Tunica Media/cytologyABSTRACT
Mucoepidermoid carcinoma (MEC) is common in the salivary glands, but alterations of the p16(INK4a) tumour suppressor gene are largely unknown. The aim of this study was to analyse p16(INK4a) gene alterations in MEC, and evaluate their significance for carcinogenesis. Thirty-eight salivary glands with MEC and six normal salivary glands were studied for p16(INK4a) alterations. In the MEC-affected group, there were 23.7% (9/38) and 13.2% (5/38) cases of homozygous deletion, and 5.3% (2/38) and 2.6% (1/38) cases of point mutation in p16(INK4a) exon 1 and exon 2, respectively. Hypermethylation of the p16(INK4a) gene promoter was found in 13 cases (13/38, 34.2%). Alterations of the p16(INK4a) gene were not found in the normal salivary glands. These findings suggest that the main mechanisms of inactivation of the p16(INK4a) gene in MEC of the salivary glands are promoter hypermethylation and homozygous deletion.
Subject(s)
Carcinoma, Mucoepidermoid/genetics , Genes, p16/physiology , Mutation/genetics , Salivary Gland Neoplasms/genetics , Adolescent , Adult , Aged , Child , Exons/genetics , Female , Gene Deletion , Gene Expression Regulation, Neoplastic/genetics , Homozygote , Humans , Male , Methylation , Middle Aged , Point Mutation/genetics , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Promoter Regions, Genetic/genetics , Salivary Glands/pathologyABSTRACT
Tartrate-resistant acid phosphatase (TRAP) isoform 5b is a potential serum marker for osteoclastic activity. Biochemical assays for serum TRAP activity with para-nitrophenylphosphate (pNPP) have low specificity for bone because of hydrolysis by unrelated nontype 5 TRAPs of blood cells and by related isoform 5a. Our purpose was to increase the specificity of TRAP assay for osteoclastic activity by using naphthol-ASBI phosphate (N-ASBI-P) as a substrate for serum type 5 TRAP activity and heparin as an inhibitor of isoform 5a. TRAP activity in individual and pooled sera of normal subjects and patients with endstage renal disease (ESRD) and rheumatologic diseases was quantitated using pNPP and N-ASBI-P as substrate at pH 5.5 and 6.1. For some experiments, heparin (23U/ml) was added as a specific inhibitor of isoform 5a activity. Isoforms 5a and 5b were separated from serum pools by cation exchange chromatography and identified by nondenaturing polyacrylamide gel electrophoresis (PAGE). N-ASBI-P was selectively hydrolyzed by TRAP isoform 5b. TRAP assays with pNPP and N-ASBI-P correlated only in ESRD sera, which contained primarily isoform 5b. The two assays did not correlate in normal or rheumatic sera with significant amounts of 5a. Heparin inhibited isoform 5a activity approximately 50% but had little effect on isoform 5b activity. Biochemical assay of serum TRAP activity can be made specific for isoform 5b by using N-ASBI-P and heparin. This method can be adapted to simple microplate biochemical or immunochemical assays. This simplified method for assessment of osteoclastic TRAP 5b activity warrants a detailed investigation in diseases of bone metabolism.
Subject(s)
Acid Phosphatase/metabolism , Bone Remodeling , Clinical Enzyme Tests/methods , Isoenzymes/metabolism , Organophosphorus Compounds/metabolism , Osteolysis/diagnosis , Acid Phosphatase/antagonists & inhibitors , Acid Phosphatase/isolation & purification , Aniline Compounds/metabolism , Arthritis, Rheumatoid/enzymology , Biomarkers , Chromatography, Ion Exchange , Electrophoresis, Polyacrylamide Gel , Enzyme Inhibitors/pharmacology , Heparin/pharmacology , Humans , Hydrogen-Ion Concentration , Hydrolysis , Isoenzymes/antagonists & inhibitors , Isoenzymes/isolation & purification , Kidney Failure, Chronic/enzymology , Osteoclasts/metabolism , Osteolysis/blood , Osteolysis/enzymology , Sensitivity and Specificity , Substrate Specificity , Tartrate-Resistant Acid PhosphataseABSTRACT
BACKGROUND: Tartrate-resistant acid phosphatase (AcP) 5b is a marker of osteoclastic activity and bone resorption. Immunoassays for serum TRAcP may lack sensitivity and specificity because of the presence of non-bone isoform 5a. The purpose of this study was to isolate the serum isoforms, quantify their disease-related expressions, and test an improved immunoassay for TRAcP 5b. METHODS: We separated TRAcP isoforms chromatographically from pooled sera of healthy, rheumatoid arthritis (RA) and endstage renal disease (ESRD) subjects. TRAcP isoforms were identified by electrophoresis and quantified by biochemical and immunochemical assays. Serum TRAcP activity in healthy, RA, and ESRD cohorts was assessed at pH 5.5 and 6.1, and compared with bone alkaline phosphatase (BAP) and N-telopeptides of type I collagen (NTx). RESULTS: TRAcP isoforms 5a and 5b were present in all sera; 5b was identical to osteoclastic TRAcP. In serum from healthy subjects, 5a accounted for 87% of the enzyme protein but only 55% of the activity. In RA, both isoforms were increased two- to threefold in protein, but their specific activities were subnormal. In ESRD, only 5b was abnormal, being increased fivefold in protein and threefold in activity. In RA sera, TRAcP activity did not correlate with either BAP or NTx. In ESRD sera, TRAcP activity correlated with BAP and NTx only when measured at pH 6.1. CONCLUSIONS: All sera contained both TRAcP isoforms 5a and 5b, but only 5b was present in bone. TRAcP isoform expression was variable in different diseases. Measurement of TRAcP activity at pH 6.1 improves the specificity of immunoassay for isoform 5b.
Subject(s)
Acid Phosphatase/metabolism , Isoenzymes/metabolism , Osteoclasts/enzymology , Arthritis, Rheumatoid/enzymology , Biomarkers/blood , Humans , Immunoassay , Kidney Failure, Chronic/enzymology , Reference Values , Sensitivity and Specificity , Tartrate-Resistant Acid PhosphataseABSTRACT
OBJECTIVE: To manufacture adriamycin-porous tricalcium phosphate (A-PTCP) ceramic drug delivery system (DDS) as a possible method for bone defect treatment after bone tumor operation. METHODS: A-PTCP DDS was made from putting adriamycin into PTCP. Thirty rabbits were divided randomly into group A(24 rabbits) and group B(6 rabbits). A-PTCP was implanted in the greater trochanter of the right femur in group A. Adriamycin were injected into veins in group B. Muscle around A-PTCP and plasma were taken out at different period. Adriamycin concentrations in muscle and plasma were measured by high performance liquid chromatography (HPLC). RESULTS: A-PTCP could gradually release adriamycin over 10 weeks. Adriamycin concentrations in the muscle were higher than that in plasma. CONCLUSION: A-PTCP may be a new method for repairing bone defects after bone tumor operation.
Subject(s)
Calcium Phosphates , Ceramics , Doxorubicin/pharmacokinetics , Drug Delivery Systems , Prostheses and Implants , Animals , Biocompatible Materials , Bone Neoplasms/metabolism , Doxorubicin/administration & dosage , Femur/surgery , Muscles/metabolism , Rabbits , Random AllocationABSTRACT
OBJECTIVE: To study the curative effect of pulpotomy in the treatment of deciduous teeth with deep caries. METHODS: 45 deciduous molars with deep caries from 3 8 years old children were selected at random. Pulpotomy was performed on these treated teeth. Clinical examination and X-ray films for the treated teeth were taken to follow up all the teeth at 6 months, one year and two years respectively after treatment. RESULTS: At one year after treatment, 45 cases were followed up, the rate of success in clinical examination and in X-ray photographs both were 100%. While at two years after treatment, 42 cases were followed up and 3 cases were lost, the rate of success in clinical examination was 95% and the rate of success in X-ray photographs was 88%. The reasons of the failure were internal absorption and pulpitis followed by fracture and exfoliation of the obturators in the treated teeth. CONCLUSION: Pulpotomy in deciduous teeth with deep caries was a reliable and effective method. It is necessary to follow up the treated teeth by X-ray at regular intervals after treatment.
ABSTRACT
The objective of this study was to identify the isoform, type-5a or type-5b, responsible for increased tartrate-resistant acid phosphatase (TRAP) activity in endstage renal disease (ESRD) and TRAP protein in rheumatoid arthritis (RA). We studied 24 sera each from healthy, ESRD and RA subjects. Type-5 TRAP activity and protein were quantitated by immunoassays. Isoform expression was determined by computerized imaging of non-denaturing polyacrylamide gels (PAGE) stained for TRAP activity. Other biochemical markers included: intact parathyroid hormone (iPTH), total and bone-specific alkaline phosphatase (TAP, BAP), N-telopeptides of type-I collagen (NTx), and free pyridinoline (Pyd). Isoform 5a was normal in both ESRD and RA. Isoform 5b was elevated in ESRD only. Serum TRAP activity correlated with both isoforms 5a and 5b in RA, but only with 5b in ESRD. TRAP protein assays did not correlate with PAGE assays for 5a or 5b. TRAP activity, but not protein, correlated with BAP and NTx in RA sera. Both TRAP activity and protein correlated with iPTH, TAP and Pyd in ESRD sera. Increased TRAP activity in ESRD was due to increased osteoclastic isoform 5b and related to bone turnover. Increased TRAP protein in RA was suspected, but not proven, to be isoform 5a and not related to bone turnover. Heterogeneity of serum TRAP and preferential expression of isoforms has clinical significance in different diseases including ESRD and RA.
Subject(s)
Acid Phosphatase/blood , Arthritis, Rheumatoid/blood , Isoenzymes/blood , Kidney Failure, Chronic/blood , Bone and Bones/metabolism , Electrophoresis, Polyacrylamide Gel , Humans , Tartrate-Resistant Acid PhosphataseABSTRACT
OBJECTIVE:To observe the effect of guided tissue regeneration (GTR) on induction of ostegenesis in developmental bone clefts,and provide a basis for the use of GTR to repair bony clefts in cleft lip and palate patients.METHODS:The e-PTFE membrane was placed on the labial and lingual sides of the rabbit mandibular central fissure.A radiological and pathological study was performed to determine the healing of the fissure integration.RESULTS:The results showed that the bony cleft was completely integrated 4 weeks after placement of the barrier membrane.CONCLUSION:GTR is an effective approach to reconstruct bony cleft,and is likely to be used in repair of alveolar cleft and cleft palate.
ABSTRACT
The magnetic porous tricalcium phosphate (MPTCP) and porous tricalcium phosphate (PTCP) ceramic cylinders were implanted into right and left bone defects of rabbits' radii in order to determine the utility of the MPTCP ceramics. Based on naked eye inspection, light and scanning electron microphotography, roentgenography, quantitative histological measurement of new bone formation and anti-break test for a period of 5 months. The results showed that the two kinds of ceramics were biocompatible with human tissue. MPTCP ceramics could induce more new bone formation than PTCP ceramics. Treatment of fractures with synthetic calcium phosphate ceramics and magnetic fields were discussed.