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Background@#Advanced technology has become a valuable tool in etiological studies of intellectual disability/global developmental delay (ID/GDD). The present study investigated the role of genetic analysis to confirm the etiology in ID/GDD patients where the cause of the disease was uncertain in central China.@*Methods@#We evaluated 1051 ID/GDD children aged 6 months to 18 years from March 2009 to April 2017. Data concerning basic clinical manifestations were collected, and the method of etiology confirmation was recorded. Genome-wide copy number variations (CNVs) detection and high-throughput sequencing of exons in the targeted regions was performed to identify genetically-based etiologies. We compared the incidence of different methods used to confirm ID/GDD etiology among groups with differing degrees of ID/GDD using the Chi-square or Fisher exact probability test.@*Results@#We recruited 1051 children with mild (367, 34.9%), moderate (301, 28.6%), severe (310, 29.5%), and profoundly severe (73, 6.9%) ID/GDD. The main causes of ID/GDD in the children assessed were perinatal factors, such as acquired brain injury, as well as single gene imbalance and chromosomal gene mutation. We identified karyotype and/or CNVs variation in 46/96 (47.9%) of cases in severe ID/GDD patients, which was significantly higher than those with mild and moderate ID/GDD of 34/96 (35.4%) and 15/96 (15.6%), respectively. A total of 331/536 (61.8%) patients with clear etiology have undergone genetic analysis while 262/515 (50.9%) patients with unclear etiology have undergone genetic analysis (χ2 = 12.645, P < 0.001). Gene structure variation via karyotype analysis and CNV detection increased the proportion of children with confirmed etiology from 51.0% to 56.3%, and second-generation high-throughput sequencing dramatically increased this to 78.9%. Ten novel mutations were detected, recessive mutations in X-linked genes (ATPase copper transporting alpha and bromodomain and WD repeat domain containing 3) and dominant de novo heterozygous mutations in X-linked genes (cyclin-dependent kinase like 5, protocadherin 19, IQ motif and Sec7 domain 2, and methyl-CpG binding protein 2) were reported in the study.@*Conclusions@#The present study indicates that genetic analysis is an effective method to increase the proportion of confirmed etiology in ID/GDD children and is highly recommended, especially in ID/GDD children with uncertain etiology.
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BACKGROUND@#Advanced technology has become a valuable tool in etiological studies of intellectual disability/global developmental delay (ID/GDD). The present study investigated the role of genetic analysis to confirm the etiology in ID/GDD patients where the cause of the disease was uncertain in central China.@*METHODS@#We evaluated 1051 ID/GDD children aged 6 months to 18 years from March 2009 to April 2017. Data concerning basic clinical manifestations were collected, and the method of etiology confirmation was recorded. Genome-wide copy number variations (CNVs) detection and high-throughput sequencing of exons in the targeted regions was performed to identify genetically-based etiologies. We compared the incidence of different methods used to confirm ID/GDD etiology among groups with differing degrees of ID/GDD using the Chi-square or Fisher exact probability test.@*RESULTS@#We recruited 1051 children with mild (367, 34.9%), moderate (301, 28.6%), severe (310, 29.5%), and profoundly severe (73, 6.9%) ID/GDD. The main causes of ID/GDD in the children assessed were perinatal factors, such as acquired brain injury, as well as single gene imbalance and chromosomal gene mutation. We identified karyotype and/or CNVs variation in 46/96 (47.9%) of cases in severe ID/GDD patients, which was significantly higher than those with mild and moderate ID/GDD of 34/96 (35.4%) and 15/96 (15.6%), respectively. A total of 331/536 (61.8%) patients with clear etiology have undergone genetic analysis while 262/515 (50.9%) patients with unclear etiology have undergone genetic analysis (χ = 12.645, P < 0.001). Gene structure variation via karyotype analysis and CNV detection increased the proportion of children with confirmed etiology from 51.0% to 56.3%, and second-generation high-throughput sequencing dramatically increased this to 78.9%. Ten novel mutations were detected, recessive mutations in X-linked genes (ATPase copper transporting alpha and bromodomain and WD repeat domain containing 3) and dominant de novo heterozygous mutations in X-linked genes (cyclin-dependent kinase like 5, protocadherin 19, IQ motif and Sec7 domain 2, and methyl-CpG binding protein 2) were reported in the study.@*CONCLUSIONS@#The present study indicates that genetic analysis is an effective method to increase the proportion of confirmed etiology in ID/GDD children and is highly recommended, especially in ID/GDD children with uncertain etiology.
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@# Objective To compare performance of C5.0 decision tree models and radial basis function(RBF) neural network in predicting the risk of hemorrhagic transformation in acute ischemic stroke. Methods Patients with acute ischemic stroke admitted to hospital were enrolled. Hemorrhagic transformation group and non-hemorrhagic transformation group were divided according to whether hemorrhagic transformation occurred within 2 weeks after admission. Retrospectively collected patients’ case information. C5.0 decision tree models and RBF neural network model were established with the ratio of 7 :3 for training set and test set, and the prediction performance of the model was compared. Results A total of 460 patients’ case information were collected and divided in 314 training set samples and 146 test set samples. Accuracy rates of the C5.0 decision tree model were 96.5% and 80.1%, sensitivities were 98.1% and 82.6%, specificities were 94.8% and 77.9%, Kappa index were 0.93 and 0.60, and AUC were 0.97 and 0.80. Accuracy rates of the neural network model were 72.6% and 74.7%, sensitivities were 87.6% and 88.4%, specificities were 56.9% and 62.3%, Kappa index were 0.45 and 0.50, and AUCs were 0.72 and 0.75. In the training set, the prediction performance of the C5.0 decision tree model was superior to the RBF neural network model. However, there was no statistical difference in the test set.Conclusion C5.0 decision tree model is better than RBF neural network model in risk prediction.
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<p><b>OBJECTIVE</b>To explore the ratio of lymphocyte subsets in peripheral blood of patients with aplastic anemia (AA) and patients with hypoplastic myelodysplastic syndrome (hypo-MDS) patients and to evaluate their significance.</p><p><b>METHODS</b>The clinical data of 181 cases of AA and 111 cases of hypo-MDS from January 2008 to December 2014 were collected from Blood Diseases Hospital of Chinese academy of medical sciences, and then the differences of lymphocyte subsets and its effect in 2 groups were analyzed.</p><p><b>RESULTS</b>CD4/CD8ratio, proportion of CD3cells and its subsets CD3CD4/CD3CD8cells in hypo-MDS group were not significant different from AA group (P>0.05). the proportion of CD3CD16/CD56NK cells and CD3CD57T-LGL cells in hypo-MDS group was significantly higher than that in AA group (P<0.05, P<0.01), but CD19B lymphocyte percentage in hypo-MDS patients was lower than that in AA patients (P<0.05). After dividing group according to CD4/CD8ratio, the ratios of CD3CD16/CD56NK cells and CD3/CD57T-LGL cells were higher only in normal CD4/CD8ratio group of hypo-MDS patients than those in AA patients, while the ratio of B lymphocytes was significant different in inverted CD4/CD8ratio group between hypo-MDS and AA patients. The CD19B lymphocyte ratio in hypo-MDS patients was significantly lower than that in AA patients (P<0.05). As well, the levels of erythrocytes and platelets in peripheral blood between hypo-MDS and AA patients only in normal CD4/CD8ratio group were significantly different, while the significant difference of WBC count and reticulocyte ratio were observed in high CD4/CD8ratio and non-inverted CD4/CD8ratio groups, respectively; the significant difference of bone marrow blast ratio and muture monocyte ratio was found in high CD4/CD8ratio group.</p><p><b>CONCLUSION</b>The changes of lymphocyte subsets can be used as an reference indicator for differential diagnosis of hypo-MDS and AA. The comparative analysis of patients with these 2 kinds of diseases after dividing into subgroups according to ratio of CD4/CD8cells is beneficial to differentiat diagnosis.</p>
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<p><b>OBJECTIVE</b>To investigate the clinical and electrophysiological characteristics and prognosis of acute motor axonal neuropathy (AMAN) in children in South China.</p><p><b>METHODS</b>The clinical and electrophysiological data of 6 children with AMAN was analyzed, and they were followed up.</p><p><b>RESULTS</b>The mean age of onset was 4.4 years. Most patients came from rural areas and 5 cases had a history of prodromal infection. There were no seasonal differences in clinical onset among the patients. The most common first symptom was muscle weakness, and the mean time from onset to the most severe disease status was 4.2 days. Nerve conduction test results revealed that all patients showed significantly lower amplitude of motor nerve action potential, only 22.3%-73.4% of the lower limit of normal. Injury to the nerves of distal extremities was more serious than injury to the nerves of proximal extremities (P<0.05), while there was no significant difference in the injury to the nerves of upper and lower extremities (P>0.05). Motor nerve conduction velocity and sensory nerve conduction velocity were normal. All patients received intravenous immunoglobulin (IVIG). Of the 6 AMAN patients, 4 could walk independently after a follow-up of 3 months to 1 year.</p><p><b>CONCLUSIONS</b>AMAN in children occurs mostly in rural areas. There is no seasonal difference in the clinical onset of the disease. Muscle weakness is the most common first symptom and the worst status of AMAN appears in the early stage of the disease. Electrophysiological examination provides important information for the diagnosis of AMAN. Some children with AMAN regain the ability to walk independently 1 year after onset. Early application of IVIG treatment may help recovery of neural function.</p>
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Child , Child, Preschool , Female , Humans , Infant , Male , Guillain-Barre Syndrome , Drug Therapy , Allergy and Immunology , Immunoglobulins, Intravenous , Therapeutic Uses , Neural Conduction , Physiology , PrognosisABSTRACT
<p><b>OBJECTIVE</b>To observe and compare the effects of aging and hypertension on rats' aortic vascular smooth muscle cells (VSMCs) and the effects of the extracts from Panax ginseng, Panax notoginseng and Ligusticum chuanxiong.</p><p><b>METHODS</b>The rat aortic VSMCs model was established using the method of primary cell culture. Of them, the rats of the aging experiment were divided into 5 groups, i.e., the young control group (as Yon), the old group (as Old), the old + probucol group (as Old+Pro), the old +low dose extracts group (as Old+Pro), and the old+high dose extracts group (as Old+High). The rats of the hypertension experiment were divided into 5 groups, i.e., the Wistar-Kyoto control group (as WKY), the spontaneously hypertensive rat group (as SHR), the SHR +Valsartan group (as SHR+Val), the SHR+low dose extracts group (as SHR+Low), and the SHR+high dose extracts group (SHR+High). The proliferation of VSMCs was detected using MTT. The expression of MMP-9 was detected by immunocytochemical assay. The mRNA and protein expressions of peroxisome proliferator activated receptor-gamma (PPAR-gamma) were detected using RT-PCR and Western blot respectively.</p><p><b>RESULTS</b>Compared with the Yon group, the proliferation of VSMCs and the MMP-9 expression increased, the mRNA and protein expressions of PPAR-gamma decreased in the Old group, all showing statistical difference (P < 0.05). Compared with the Old group, the proliferation of VSMCs and the MMP-9 expression obviously decreased, the mRNA expression of PPAR-gamma obviously increased in the Old+Pro group, the Old+High group, and the Old+Low group (all P < 0.05). The PPAR-y protein expression obviously increased in the Old+Pro group and the Old+Low group (P < 0.05). Compared with the WKY group, the proliferation of VSMCs and the expression of MMP-9 obviously increased, the mRNA and protein expressions of PPAR-gamma obviously decreased in the SHR group (all P < 0.05). Compared with the SHR group, the proliferation of VSMCs and the expression of MMP-9 obviously decreased, the mRNA and protein expressions of PPAR-gamma obviously increased in the SHR+Val group, the SHR+High group, and the SHR + Low group (all P < 0.05).</p><p><b>CONCLUSIONS</b>Both aging and hypertension could result in excessive proliferation of rat aortic VSMCs and the expression changes of correlated cytoactive factors. The extracts from Panax ginseng, Panax notoginseng (Burk.) and Ligusticum chuanxiong can lower their proliferation levels and reduce the expressions of negative cytokines, thus reducing aging and hypertension induced injury of VSMCs and delaying angiocellular aging.</p>
Subject(s)
Animals , Male , Rats , Aging , Drugs, Chinese Herbal , Pharmacology , Ligusticum , Matrix Metalloproteinase 9 , Metabolism , Muscle, Smooth, Vascular , Cell Biology , Metabolism , Myocytes, Smooth Muscle , Metabolism , PPAR gamma , Metabolism , Panax , Panax notoginseng , Plant Extracts , Pharmacology , RNA, Messenger , Genetics , Rats, Inbred SHR , Rats, Inbred WKY , Rats, WistarABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of extracts from Radix Ginseng, Radix Notoginseng and Rhizoma Chuanxiong (EXT) on delaying vascular smooth muscle cells (VSMCs) aging in aged rats.</p><p><b>METHODS</b>VSMCs were obtained by the modified tissue explants technique and were shown to be positive for smooth muscle α-actin (SM-α-actin) by immunohistochemistry staining. VSMCs obtained from the young rats were served as the young control group; VSMCs obtained from the old rats were treated with no drug (the old group), with low dose extracts (20 mg/L, the EXT low-concentration group) and high dose extracts (40 mg/L, the EXT high concentration group), and with Probucal (10(-6) mol/L, the Probucal group) as a positive control. All groups were cultured for 24 h in the medium with 10% serum for 24 h followed by another 24 h in the serum-free medium. At the end of the 48-h culture, the following analyses were performed including determination of senescence-associated β-galactosidase (SAβ-Gal) activity, flow cytometry analysis of cell cycle, real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) analyses of p16, Cyclin D1, cyclin-dependent kinase 4 (CDK4) and retinoblastoma (Rb) mRNA expression, and Western blotting analyses of p16, cyclin D1, CDK4 and phosphoretinoblastoma (pRb) protein expressions.</p><p><b>RESULTS</b>(1) In comparison to the younger rats, VSMCs from aged rats had significantly more SAβ-Gal positive cells (P<0.01) and more cells in S phase (P<0.05). VSMCs from the all treated groups showed a significant decrease in both SAβ-Gal positive cells (P<0.05) and S phase (P<0.05) compared to the old rats. (2) Compared with the young group, VSMCs in the old group had a significant decrease in p16 and Rb mRNA expression and a significant increase in Cyclin D1 and CDK4 mRNA expression. Compared with the old group, VSMCs in the treated groups had a significant increase in p16 and Rb mRNA expression and a significant decrease in Cyclin D1 and CDK4 mRNA expression (P<0.05). (3) Compared with the young group, VSMCs in the old group had a significant decrease in p16 protein expression and a significant increase in Cyclin D1, CDK4 and pRb protein expressions (P<0.05). Compared with the old group, VSMCs in the treated groups had a significant increase in p16 protein expression and a significant decrease in cyclinD1, CDK4 and pRb protein expressions (P<0.05).</p><p><b>CONCLUSIONS</b>VSMCs obtained from old rats showed typical signs of cellular senescence and vascular aging. EXT had an effect on delaying senescence of VSMCs in vitro by altering the p16-cyclinD/CDK-Rb pathway.</p>
Subject(s)
Animals , Male , Rats , Aging , Aorta , Cell Biology , Cell Cycle , Cellular Senescence , Cyclin D1 , Genetics , Metabolism , Cyclin-Dependent Kinase 4 , Genetics , Metabolism , Cyclin-Dependent Kinase Inhibitor p16 , Genetics , Metabolism , Drugs, Chinese Herbal , Pharmacology , Flow Cytometry , Gene Expression Regulation , Muscle, Smooth, Vascular , Cell Biology , Myocytes, Smooth Muscle , Cell Biology , Metabolism , Panax , Plant Extracts , Pharmacology , RNA, Messenger , Genetics , Metabolism , Rats, Wistar , Retinoblastoma Protein , Genetics , Metabolism , beta-Galactosidase , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the influence of lipopolysaccharide (LPS) on the permeability of rat brain microvascular endothelial cells (BMECs) and possible molecular mechanism.</p><p><b>METHODS</b>Monolayers of primary rat BMECs were separated and cultured, and then treated with (LPS group) or without LPS (control group). The barrier integrity was measured by transendothelial electrical resistance (TEER) assay. The degrees of RhoA activation were determined by Pull-down assay. The expression levels of p115RhoGEF, zonula occludens-1 (ZO-1), occludin and claudin-5 proteins were detected by Western blot analysis.</p><p><b>RESULTS</b>The average TEER values of rat BMECs in the LPS group were 108.3±4.2 Ω•cm2 and 85.4±2.5 Ω•cm2 respectively 3 and 12 hrs after LPS treatment, which were significantly lower than that in the control group (159.0±8.6 Ω•cm2). Compared with the control group, the activity of RhoA started to increase 5 minutes after LPS treatment, and the expression of p115RhoGEF protein started to increase 1 hr after LPS treatment and the cellular protein levels of ZO-1, occludin and claudin-5 decreased significantly 3 hrs after LPS treatment in the LPS group (P<0.05).</p><p><b>CONCLUSIONS</b>LPS may activate the p115RhoGEF/RhoA pathway and decrease protein expression of ZO-1, occludin and claudin-5, resulting in an increased permeability of rat BMECs.</p>
Subject(s)
Animals , Rats , Brain , Capillary Permeability , Electric Impedance , Endothelial Cells , Metabolism , Guanine Nucleotide Exchange Factors , Lipopolysaccharides , Pharmacology , Rats, Sprague-Dawley , Rho Guanine Nucleotide Exchange Factors , Tight Junctions , Chemistry , rhoA GTP-Binding ProteinABSTRACT
<p><b>BACKGROUND</b>Psoriasis is a common inflammatory skin disease, yet knowledge of the factors that may induce, trigger, or exacerbate psoriasis is not fully delineated. Recent advances have improved our understanding of the link between psoriasis and cell-wall-deficient bacteria (CWDB) infections. In the present study we assessed the prevalence of CWDB infection in patients with psoriasis.</p><p><b>METHODS</b>The carriage rate of CWDB in the tonsil or pharynx of psoriasis patients, chronic tonsillitis patients and controls were investigated using hypertonic medium. Psoriasis patients with CWDB were randomly assigned to two groups and respectively treated with antibiotics or systemic therapy without antibiotic. Human peripheral blood mononuclear cells (PBMC) from psoriasis patients, chronic tonsillitis patients and control subjects were stimulated with bacteria antigens and extra-cellular levels of interferon-gamma (IFN-gamma) and interleukin (IL)-10 were measured in the supernatants using the ELISA technique, in vitro. Meanwhile, the proliferation ability of PBMC to respond to bacteria antigens was detected by MTT assay.</p><p><b>RESULTS</b>CWDB were isolated from 74.2% of psoriasis patients, 23.5% of chronic tonsillitis patients and only 6.3% of controls. Antibiotic therapy was appropriate for approximately 80% of psoriasis patients with CWDB infection, and in only 8.9% psoriasis patients CWDB infection was detected after antibiotic therapy. Meanwhile, our study showed that CWDB and wide-type bacteria did remarkably enhance the production of IFN-gamma, in vitro, and PBMC proliferation.</p><p><b>CONCLUSION</b>CWDB infection may be a virtual triggering factor in psoriasis by regulating T-cell activation.</p>