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Objective To evaluate the reliability of sweep visual evoked potential (SVEP) for assessing the normal objective visual acuity in adult wild-type C57BL/6J mice.Methods A total of 6 wild C57BL/6J mice of either gender at 32-33 days of age were collected.After the mice were anesthetized,the recording electrode (a 2-mm diameter tungsten electrode) was placed at 3 mm lateral to lambda over the left cortex (contralateral to the stimulated right eyes) and advanced to 400 μm within the cortex,and the needle electrodes which placed in the forehead and tail served as the reference and the ground,accordingly.Left eyes (unstimulated eyes) were covered during the recording.The screen (mean luminance 25 cd · m-2) was placed 20 cm in front of the mice,thereby covering 100° (horizontal) × 82° (vertical) of the visual field.The mice were stimulated by vertically sinusoidal grating,which consisted of 11 spatial frequencies,stimulus contrast was 100%,and time frequency was 1 Hz;the entire sweep took 12 seconds (11 spatial frequencies + 1s pre-adaptation),and the values were averaged until the error bars for the data were stable.The VEP waveforms of visual cortex corresponding to the left and right eyes were recorded separately.At least 3-4 trials consisting of 20 events for different frequencies were averaged.Left and right eye were tested alternately.Then the coordinate system was established by using the spatial frequency logarithm as x-axis and the amplitude of the response obtained by the discrete Fourier analysis as y-axis,the objective visual acuity was obtained by extrapolation to zero amplitude of the linear regression through the 4 effective data points adjacent to noise.Results SVEP showed that the amplitude of N100 wave was maximal at a spatial frequency of 0.05 c · deg-1.With the increase of the stimulus spatial frequency progressively,SVEP decreased in amplitude and increased in latency.The cortical electrical signal could not be distinguished from noise at a spatial frequency of 0.65 c · deg-1.The average acuity of 6 mice was (0.56 ± 0.04) c · deg-1 for right eyes and (0.50 ± 0.04)c · deg-1 for left eyes respectively.There was no significant difference between the two eyes(P>0.05);The visual evoked potential acuity of 12 eyes was (0.53 ±0.03)c ·deg-1.Conclusion SVEP could be used to estimate the mouse VEP acuity,which was closed to the behavioral visual acuity.However,the VEP acuity was more objective and accurate as well as time-saving.
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<p><b>OBJECTIVE</b>To investigate the association of serum vitamin D [25-(OH)D] level with the severity and treatment in children with Henoch-Schönlein purpura (HSP).</p><p><b>METHODS</b>A total of 50 children with newly-diagnosed HSP between January and December, 2015 were enrolled as HSP group, and 49 healthy children were enrolled as control group. Fasting serum samples were collected, and ELISA was used to measure serum 25-(OH)Dlevel. According to the serum 25-(OH)Dlevel, the HSP group were further divided into normal group (>20 ng/mL) (n=9), insufficiency group (15-20 ng/mL) (n=15), deficiency group (≤15 ng/mL) (n=25), and severe deficiency group (≤5 ng/mL) (n=1). The general data, clinical manifestations, hormone therapy, course of disease before admission, and length of hospital stay were compared between groups.</p><p><b>RESULTS</b>The HSP group had a significantly lower serum 25-(OH)Dlevel than the control group (16±6 ng/mL vs 29±5 ng/mL; P<0.01). Compared with the normal and insufficiency groups, the deficiency and severe deficiency groups had significant increases in the incidence rate of renal involvement, rate of hormone application, and median length of hospital stay (P<0.05), while there was no significant difference in course of disease before admission (P>0.05).</p><p><b>CONCLUSIONS</b>Children with HSP have a low serum 25-(OH)Dlevel, and such children may have a high risk of renal involvement, a high rate of hormone application, and a prolonged length of hospital stay. However, further studies are needed to investigate whether vitamin D supplementation is helpful to the treatment of HSP and can shorten the course of disease in children with HSP.</p>
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Criança , Feminino , Humanos , Masculino , Tempo de Internação , Vasculite por IgA , Sangue , Tratamento Farmacológico , Índice de Gravidade de Doença , Vitamina D , SangueRESUMO
<p><b>BACKGROUND</b>The mechanisms of pathological retinal neovascularization (RNV) remain unknown. Several microRNAs were reported to be involved in the process of RNV. Oxygen-induced retinopathy (OIR) is a useful model to investigate RNV. Our present work explored the expression and the role of microRNA-128 (miR-218) in oxygen-induced RNV.</p><p><b>METHODS</b>OIR was used to establish RNV model. The expression level of miR-218 in the retina from OIR mice was assessed by quantitative real-time reverse transcriptase polymerase chain reaction. Fluorescein angiography was performed in retinae of OIR mice, and RNV was quantified by hematoxylin and eosin staining to evaluate the effect of pCDH-CMV-miR-218 intravitreal injection on RNV in OIR mice. Roundabout 1 (Robo1) expression was detected by Western blotting in mouse retinal vascular endothelial cells expressing a high or low level of miR-218 and retinal tissues from OIR mice. Cell migration was evaluated by scratch wound assay.</p><p><b>RESULTS</b>In OIR mice, the expression level of miR-218 was significantly down-regulated (P = 0.006). Retinal Robo1 expression was significantly increased at both mRNA and protein levels (P = 0.001, 0.008; respectively). miR-218 intravitreal injection inhibited retinal angiogenesis in OIR mice, and the restoration of miR-218 in retina led to down-regulation of Robo1.</p><p><b>CONCLUSIONS</b>Our experiments showed that restoration of miR-218 inhibited retinal angiogenesis via targeting Robo1. MiR-218 contributed to the inhibition of retinal angiogenesis and miR-218 might be a new therapeutic target for preventing RNV.</p>
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Animais , Camundongos , Movimento Celular , Células Cultivadas , Camundongos Endogâmicos C57BL , MicroRNAs , Fisiologia , Proteínas do Tecido Nervoso , Fisiologia , Oxigênio , Farmacologia , Receptores Imunológicos , Fisiologia , Neovascularização RetinianaRESUMO
<p><b>BACKGROUND</b>Common variable immunodeficiency (CVID) is one of the most common symptomatic primary immunodeficiency syndromes. The purpose of this article was to broaden our knowledge about CVID for better diagnosis and treatment.</p><p><b>METHODS</b>Clinical and immunological features of 40 Chinese patients with CVID were analyzed retrospectively.</p><p><b>RESULTS</b>The median age at onset was 11-year-old (range 4-51 years). The median age at diagnosis was 14.5-year-old (range 5-66 years). The average time of delay in diagnosis was 5.3 years (range 1-41 years). The most common main complaint was fever due to infections (35 cases, 87.5%). Pneumonia (28 cases, 70%) was the most common type of infections. Bronchiectasis was present in 6 patients (15%). Autoimmune disease was detected in 6 cases of CVID, and malignancy in 2 cases. The median total serum levels of IgG, IgA, and IgM at diagnosis were 1.07 g/L, 0.07 g/L, and 0.28 g/L, respectively. The percentages of CD3- /CD19 + B-cells were 1%-3.14%.</p><p><b>CONCLUSIONS</b>Infection is the most frequent presentation of CVID. Patients with unexplainable infections should receive further examination including serum immunoglobulin (Ig) and lymphocyte subset analysis. Regular and sufficient substitution with Ig is recommended.</p>
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Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Humanos , Pessoa de Meia-Idade , Adulto Jovem , Bronquiectasia , Tratamento Farmacológico , Alergia e Imunologia , Patologia , China , Imunodeficiência de Variável Comum , Tratamento Farmacológico , Alergia e Imunologia , Patologia , Imunoglobulinas , Metabolismo , Imunoglobulinas IntravenosasRESUMO
<p><b>BACKGROUND</b>The mechanism of retinal neovascularization is not understood completely. Many growth factors are involved in the process of retinal neovascularization, such as vascular endothelial growth factor (VEGF) and pigment epithelium-deprived factor (PEDF), which are the representatives of angiogenic and antiangiogenic molecules respectively. Oxygen induced retinopathy (OIR) is a useful model to investigate retinal neovascularization. The present study was conducted to investigate the feasibility of small interference RNA (siRNA) targeting VEGF gene in attenuating oxygen induced retinopathy (OIR) by regulating VEGF to PEDF ratio (VEGF/PEDF).</p><p><b>METHODS</b>In vitro, cultured EOMA cells were transfected with VEGF-siRNA (psi-HI(TM)/EGFP/VEGF siRNA) and Lipofectamine(TM) 2000 for 24, 48, and 72 hours, respectively. Expression of VEGF mRNA was evaluated by real time polymerase chain reaction (PCR) and the level of VEGF protein was analyzed by Western blotting. In vivo, OIR model mice were established, the mice (C57BL/6J) received an intra-vitreal injection of 1 µl of mixture of psi-HI(TM)/EGFP/VEGF siRNA and Lipofectamine 2000. Expressions of retinal VEGF and PEDF protein were measured by Western blotting, retinal neovascularization was observed by fluorescein angiography, and quantified.</p><p><b>RESULTS</b>In vitro psi-HI(TM)/EGFP/VEGF siRNA treatment significantly reduced VEGF mRNA and protein expression. In vivo, with decreased VEGF and VEGF-PEDF ratio, significant attenuation of neovascular tufts, avascular regions, tortuous, and dilated blood vessels were observed in the interfered animals.</p><p><b>CONCLUSIONS</b>VEGF plays an important role in OIR, and the transfection of VEGF-siRNA can effectively downregulate VEGF expression in vivo, accompanied by the downregulation of VEGF-PEDF ratio, and simultaneous attenuation of retinal neovascularization was also observed. These findings suggest that VEGF/PEDF may serve as a potential target in the treatment of retinal neovascularization and RNA interference targeting VEGF expression, which represents a possible therapeutic strategy.</p>
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Animais , Camundongos , Proteínas do Olho , Camundongos Endogâmicos C57BL , Fatores de Crescimento Neural , RNA Interferente Pequeno , Genética , Neovascularização Retiniana , Terapêutica , Serpinas , Fator A de Crescimento do Endotélio Vascular , Genética , FisiologiaRESUMO
Background Arresting the overexpression of vascular endothelial growth factor (VEGF) will be a new approach to the inhibition of neovascularization.RNA interference (RNAi) can inhibit the expression of specific gene,and its application in eye has little interference to other gene expression.Objective This study was to investigate the effect of small interference RNA (siRNA) targeting VEGF on the expression of VEGF and retinal neovascularization in oxygen-induced retinopathy (OIR) model.Methods psi-HITM/enhanced green fluorescent protein (EGFP)/VEGF siRNA was designed and prepared in vitro.Mouse endothelioma (EOMA) were cultured in DMEM without antibiotic and divided into 5 groups.The cells were incubated in DMEM only in the blank control group;while 1 μl of LipofectamineTM 2000 + psi-HITM/EGFP,1 μl LipofectamineTM 2000 + 40,50 or 60 nmol/L of psi-HITM/EGFP/VEGF siRNA was added into DMEM in the negative control group and siRNA groups,respectively.The expression of VEGF mRNA and protein was detected by real time PCR (RT-PCR) and Western blot.The optimal effective concentration of VEGF siRNA was assessed.OIR models were established in 48 7-day-old C57BL/6J mice by raising them at an oxygen concentration of (75±2) % for 5 days and then to normal air.The mice were randomized into the model group,null vector group and VEGF siRNA group.1 μl of a mixture of psi-HITM/EGFP or VEGF siRNA (60 nmol/L) and LipofectamineTM 2000 was intravitreally injected,respectively,in the null vector group and VEGF siRNA group.The normal mice were used as the normal control group.Expression of VEGF mRNA and protein in the mouse retinas was detected by RT-PCR and Western blot,respectively,and FITC-dextran stretched retinal preparation was examined to evaluate the neovascularization,and retinal sections were examined to quantify the number of vascular endothelial cell nuclei extending beyond the internal limiting membrane (ILM).Results The in vitro transfection test showed that the expression of VEGF mRNA and protein in the EOMA cells was significantly different among the blank control group,negative control group and 40,50,60 nmol/L VEGF siRNA groups (F =148.890,P < 0.001; F =306.960,P < 0.001),and the expression of VEGF mRNA was lower in different concentrations of VEGF siRNA groups than that in the blank control group (t=73.950,119.890,156.480,all at P<0.001).Also,the expression of VEGF protein was less in different concentrations of VEGF siRNA groups than that in the blank control group (t =15.452,23.344,42.119,all at P<0.001).The optimal inhibitory concentration of VEGF siRNA was 60 nmol/L.In vivo study determined that compared to the model group and null vector group,the non-perfusion zones and neovascular net in the retina were decreased,and the number of vascular endothelial cell nuclei extending beyond the ILM was less in the VEGF siRNA group.The relative expression level of VEGF mRNA in the retinas was 1.23±0.18,4.02±0.16,3.98±0.19 and 1.98±0.12 in the normal control group,model group,null vector group and VEGF siRNA group,respectively,with a significant difference among them (F=369.780,P<0.001),and the relative expression levels of VEGF mRNA in the model group and null vector group were higher than that in the normal control group (t=37.880,37.336,both P<0.001),and the expression of VEGF mRNA in the VEGF siRNA group declined by 50.8% (t=10.183,P<0.001).The difference in the expression levels of VEGF protein also was assayed among the various groups (F=408.980,P<0.001),and VEGF level in the retina was lowered by 68.0% in the VEGF siRNA group compared to the model group (t =11.473,P<0.001).However,VEGF level in the VEGF siRNA group remained at a high level in comparison with the normal control group (t =2.422,P<0.001).Conclusions Intravitreal injection of VEGF siRNA can attenuate retinal neovascularization by effectively downregulate the expression VEGF mRNA and protein in the retina.
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Objective To investigate children with nephrotic syndrome (NS) receiving long-term glucocorticoid treatment while monitoring insulin resistance(IR) index,and analyze the influencing factors,and to observe the efficacy of metformin for preventing glucocorticoid-induced obesity and diabetes mellitus type 2.Methods A total of 32 cases including 20 boys and 12 girls with mean age of 7.6 years old(2 to 14 years old),were diagnosed as NS.All children undergoing long-term treatment of prednisone > 1 mg/(kg · d) for at least 3 months.The height,body weight and body mass index(BMI) were monitored.The concentrations of serum insulin and C-peptide were determined by radioimmunoassay,the concentrations of serum HbA1 c were determined by high performance liquid chromatography,and the levels of fasting glucose,cholesterol and triglyceride were measured with an automatic measuring analyzer (American ABBO TT CCX-Ⅱ).The homeostasis model assessment-estimated insulin resistance(HOMA-IR) was calculated;When HOMA-IR > 3.5,the patients were given metformin treatment for 12 weeks while supervising their height,weight,fasting blood glucose levels monthly.Results Out of 32 cases of NS with long-term use of prednisone,23 cases had HOMAIR > 3.5 (72%).Thus,IR group included 23 patients,and non-IR (NIR) group had 9 patients.There were significant differences between 2 groups in BMI,age,prednisone dosage and the levels of triglyceride (TG) (all P < 0.05).After 12 weeks of metformin treatment,the HOMA-IR (5.24 ± 1.82 vs 2.54 ± 1.09,P < 0.01),HbA1 c levels [(6.36 ±0.82)% vs(5.39 ±0.51)%,P <0.05],BMI[(27.42 ±6.12) kg/m2 vs(22.72 ±5.48) kg/m2,P <0.01],and TG levels[(2.03 ± 1.10) mmol/L vs(1.45 ±0.48) mmol/L,P<0.05] were significantly reduced.Conclusions Older children with long-term use of glucocorticoid may easily lead to IR.Therefore,monitoring the HOMA-IR and early administration of mefformin is helpful for prevention and treatment of glucocorticoid-induced obesity and diabetes mellitus type 2.
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<p><b>OBJECTIVE</b>To study the risk factors for recurrent pneumonia in children without underlying diseases.</p><p><b>METHODS</b>A case-control study was conducted in 106 children with recurrent pneumonia (case group) and 106 age, gender- and weight-matched children with pneumonia but no recurrence (control group). The children in both groups had no underlying disease. The risk factors for recurrent pneumonia were investigated by the Chi-Square analysis and the multivariate logistic regression model.</p><p><b>RESULTS</b>The Chi-Square analysis showed that the percentages with the history of wheezing, allergy (food or medicine) and eczema and the percentage of transient neutropenia in the case group were significantly higher than those in the control group. The multivariate logistic regression analysis showed that the wheezing history (OR=13.387, 95% CI: 5.541-32.343), allergic history (food or medicine) (OR=4.267, 95% CI: 2.081-8.751) and transient neutropenia (OR=3.606, 95% CI: 1.806-7.202) were the independent risk factors of recurrent pneumonia. CONCLUSIONS The wheezing history, allergic history and transient neutropenia may increase the risk of recurrence of pneumonia in pneumonic children without underlying diseases.</p>
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Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Estudos de Casos e Controles , Modelos Logísticos , Pneumonia , Recidiva , Fatores de RiscoRESUMO
Background About one third of congenital cataract is associated with inheriting factor.The inherited heterogeneity has been found in congenital cataract.To seek the pathogenic gene is essential for the gene therapy. Objective Present study was to map and identify the causal gene for autosomal dominant congenital cataract (ADCC) in a Chinese family. Methods The clinical features of all affected members in this family were examined.Blood samples were collected from eleven family members for genetic linkage analysis.Polymorphic microsatellite markers were selected from the regions which harbor all known loci linked with ADCC.Universal fluorescent-labeled M13 primer was used in linkage analysis.Direct genomic sequencing was used to evaluate the candidate gene for example CRYBB2 gene.This study followed Helsinki Declaration and was proved by Tianjin City Ethic Committee.Written informed consent was obtained from each SUbject before any medical procees. Results The maximum two-point LOD score of 1.20 was obtained for marker D22S315 (θ=0).The LOD score of 0.6 was obtained for marker D16S3068.No mutation in all exons of CRYBB2 gene was found in the family. Conclusion CRYBB2 gene associated with ADCC was excluded from the family.A genome-wide linkage screening should be conducted.Genotyping with microsatellite markers using Universal fluorescent-labeled M13 primer can decrease the cost and obtain the same result.
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Background Clinical and genetic heterogeneity of congenital cataract is well substantiated.Researchers often identify disease loci by linkage analysis and screen candidate gene by direct sequencing.Objective This study was to localize and identify the disease-causing genes for two Chinese families with congenital coralliform cataracts.Methods Two Chinese families(CC1 and CC2) with autosomal dominant inheritance congenital coralliform cataracts were ascertained and patients in the families underwent ophthalmological examination.Periphery blood samples were collected and DNA was extracted from 17 subjects including 11 cataract patients and 4 phenotype normal and 2 spouses.A linkage scan of genomic regions containing 25 known candidate genes was performed using 50 polymorphic microsatellite markers on genomic DNA from affected and unaffected family members and LOD scores were calculated.Candidate genes were sequenced and mutations were analyzed.Three single nucleotyde polymorphisms(SNP)(rs2305429,rs2305430,rs2242074) were sequenced and genotyped for the detect of the possibility of a common origin between CC1 and CC2.This study complied with the Declaration of Helsinki and was approved by Ethic Committee of Tianjin Eye Hospital.The informed consent was obtained from subjects and their guardian before the protocol.Results A significant LOD score of 3.28(θ=0) in family CC1 and a maximum LOD score of 1.50(θ=0) in family CC2 were both produced at the microsatellite marker D2S325 linked with CRYGD gene.Sequencing of CRYGD gene showed a heterozygous single base pair change c.70C>A in exon2,predicting to result in a P23T amino acid change.The haplotypes of two probands in their respective families was quite distinct.Conclusion These results indicate that c.C70A(p.P23T) mutation in CRYGD gene is the underlyingmolecular pathogenesis of the two families with congenital coralliform cataracts,and this mutation occurs independently in these two families rather than descending from a common ancestor.
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<p><b>OBJECTIVE</b>To assess the practical value of BIOMED-2 primers in the diagnosis of ocular adnexal lymphoma by PCR.</p><p><b>METHODS</b>DNA was extracted from 63 formalin-fixed paraffin-embedded (FFPE) ocular adnexal lymphoma specimens. The DNA quality was evaluated by PCR-based amplification of housekeeping gene beta-actin. IgH(B) and IgK(B) primers of BIOMED-2 standardized clonality analysis system were used to evaluate the immunoglobin gene rearrangements. PCR products were analyzed using capillary electrophoresis and GeneScan software.</p><p><b>RESULTS</b>76.2% (48/63) of FFPE samples produced amplifiable DNA for detection of Ig gene rearrangements.Positive detection rates by BIOMED-2 IgH(B) and IgK(B) primers were 79.2% (38/48) and 68.8% (33/48), respectively, with a combined positive detection rate of 91.7% (44/48).</p><p><b>CONCLUSIONS</b>IgH(B) and IgK(B) primers of BIOMED-2 are suitable for the detection of clonal rearrangements of Ig gene using FFPE specimens of ocular adnexal lymphomas.</p>
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Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Actinas , Genética , Primers do DNA , Neoplasias Oculares , Diagnóstico , Genética , Patologia , Rearranjo Gênico de Cadeia Pesada de Linfócito B , Rearranjo Gênico de Cadeia Leve de Linfócito B , Linfoma de Células B , Diagnóstico , Genética , Patologia , Linfoma Folicular , Diagnóstico , Genética , Patologia , Inclusão em Parafina , Projetos PilotoRESUMO
<p><b>OBJECTIVE</b>To screen the transforming growth factor, beta-induced (TGFBI) gene mutation in three Chinese families with autosomal dominant corneal dystrophy.</p><p><b>METHODS</b>Analysis of the TGFBI gene mutations was performed by direct sequencing of the whole coding regions and exon-intron boundaries of the TGFBI gene in all affected members from the three families.</p><p><b>RESULTS</b>Three kinds of TGFBI gene mutations, R124C and H626R were detected in the patients of the two lattice conneal dystrophy families, and R124H was detected in the Avellino corneal dystrophy family.</p><p><b>CONCLUSION</b>TGFBI gene mutations are the underlying molecular mechanism of the pathogenesis for corneal dystrophy. The R124 and H626 are the hot spots of TGFBI gene mutation in this disease.</p>
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Feminino , Humanos , Masculino , Povo Asiático , Genética , Distrofias Hereditárias da Córnea , Genética , Patologia , Substância Própria , Patologia , Análise Mutacional de DNA , Saúde da Família , Mutação , Linhagem , Fatores de Crescimento Transformadores , GenéticaRESUMO
<p><b>OBJECTIVE</b>To investigate the changes of the human telomerase reverse transcriptase gene (hTERT) alterative splicing pattern in gastric carcinogenesis.</p><p><b>METHODS</b>Three alternative splicing sites (alpha, beta, gamma) were selected to design primers. The expression of eight hTERT alternative splicing variants (ASVs) in normal gastric mucosa, precancerous lesions and gastric cancer was detected by semi-nested reverse transcription-polymerase chain reaction (RT-PCR). The expression of beta site-remaining ASV (beta (+) hTERT mRNA) in precancerous lesions and gastric cancer tissues was detected by SYBR green real-time RT-PCR.</p><p><b>RESULTS</b>The positive rate of alpha(+) beta(+)gamma(+) hTERT mRNA was significantly higher in gastric cancer than in precancerous lesions and normal mucosa (94.7% vs. 40.0% and 0, P<0.05). The positive rates of other ASVs were not different among the three groups. The positive rates of beta deletion ASV were 72.2% in normal mucosa, 95.0% in precancerous lesions and 100.0% in gastric cancer. The mRNA level of beta(+) hTERT was 5.49 folds higher in gastric cancer than in precancerous lesions.</p><p><b>CONCLUSION</b>The hTERT alternative splicing pattern changes during gastric carcinogenesis. The beta(+) hTERT mRNA is expressed increasingly during gastric carcinogenesis and may provide useful information for diagnosis of gastric cancer or precancerous lesions.</p>
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Humanos , Processamento Alternativo , Genética , Transformação Celular Neoplásica , Genética , Patologia , Células Cultivadas , Regulação Neoplásica da Expressão Gênica , Fisiologia , Lesões Pré-Cancerosas , RNA Mensageiro , Metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias Gástricas , Genética , Metabolismo , Patologia , Telomerase , Classificação , Genética , Metabolismo , Telômero , GenéticaRESUMO
<p><b>OBJECTIVE</b>To study the PAX6 gene mutation in a Chinese pedigree with congenital aniridia.</p><p><b>METHODS</b>Linkage analysis was performed to the Chinese family with congenital aniridia using two microsatellite markers D11S904 and D11S935. Analysis of the PAX6 gene mutation was done by direct sequencing of the whole coding region and exon-intron boundaries of the PAX6 gene in all affected and unaffected individuals in the family.</p><p><b>RESULTS</b>The significant Lod Score of 3.01 was acquired at D11S935. Direct DNA sequence analysis identified a 1080C to T change in exon 9 of the patients, resulting in an Arginine substitution by a stop codon at codon 240 of the PAX6 gene, which was absent in the unaffected individuals in the family and 100 normal controls.</p><p><b>CONCLUSION</b>Our results indicate that mutation p.Arg240Ter of the PAX6 is the genetic basis of the Chinese family with congenital aniridia.</p>
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Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Aniridia , Genética , Povo Asiático , Genética , Sequência de Bases , Códon sem Sentido , Proteínas do Olho , Genética , Proteínas de Homeodomínio , Genética , Repetições de Microssatélites , Dados de Sequência Molecular , Fator de Transcrição PAX6 , Fatores de Transcrição Box Pareados , Genética , Linhagem , Proteínas Repressoras , GenéticaRESUMO
<p><b>OBJECTIVE</b>Many studies have shown that glucocorticoids play a crucial role in the development of obesity and insulin resistance. This study investigated the therapeutic effects of long-term inhibition of glucocorticoid activity on obesity and insulin resistance.</p><p><b>METHODS</b>Four-week-old male Sprague-Dawley (SD) rats were randomly fed with a high-fat diet (fat content accounting for 20% of total calorie) (control group, n=8) or with a high-fat diet along with glycyrrhetic acid (GE, 800 mg/L), an inhibitor of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) for 24 weeks (GE-treated group, n=9). The body weights and the amount of food intake were monitored weekly and daily, respectively. After 24 weeks of GE treatment, oral glucose tolerance tests were performed. Blood glucose was measured by glucose oxidase method. The levels of plasma glucocorticoids, insulin and leptin were measured with radioimmunoassay. The levels of serum cholesterol and triglyceride were determined with an automatic measuring analyzer.</p><p><b>RESULTS</b>The food intake amount decreased significantly in the GE-treated group from 6 weeks and body weight gain was markedly less from 8 weeks after GE administration compared with the control group. After 24 weeks of treatment, the plasma levels of leptin and insulin in GE-treated rats were significantly reduced compared with the control group. The serum levels of cholesterol and triglyceride decreased markedly compared with the control group and the levels of blood glucose were significantly lower 15, 30, 60 and 120 minutes after oral glucose load in the GE-treated group compared with the control group.</p><p><b>CONCLUSIONS</b>Long-term GE treatment may contribute to resisting diet-induced obesity and insulin resistance.</p>
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Animais , Masculino , Ratos , 11-beta-Hidroxiesteroide Desidrogenases , Peso Corporal , Gorduras na Dieta , Inibidores Enzimáticos , Farmacologia , Glucocorticoides , Fisiologia , Teste de Tolerância a Glucose , Ácido Glicirretínico , Farmacologia , Usos Terapêuticos , Insulina , Sangue , Resistência à Insulina , Leptina , Sangue , Obesidade , Tratamento Farmacológico , Ratos Sprague-DawleyRESUMO
Objective To investigate the effect of RNA interference(RNAi) on inhibiting respiratory syncytial virus(RSV) replication through comparing the anti-RSV activities between pshRNA7816 and ribavirin in cell culture system.Methods The recombinated plasmid pshRNA7816 and ribavirin was added to HEp-2 cells.Methyl thiazolyl tetrazolium(MTT) assay was used to detect cytotoxicity of pshRNA7816 and ribavirin on normal HEp-2 cells and protective effects of them on RSV infected HEp-2 cells.The effects of pshRNA7816 and ribavirin on change of cytopathogenic effect(CPE) of HEp-2 cells induced by RSV infection were observed through microscopically.Results pshRNA7816 had not significant toxicity on the growth of HEp-2 cells,but the ribavirin had significant toxicity when the concentration above 1.0 mmol/L.The pshRNA7816 and ribavirin could alleviate the CPE of HEp-2 cells induced by RSV infection,but the pshRNA7816 showed a more potent inhibition than ribavirin.The inhibition rates of pshRNA7816 were significantly higher than the maximum inhibition rate of ribavirin on RSV infection(P
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Objective To investigate the distribution of blood pressure and incidence of hypertension in children and adolescents from the urban area of Guiyang city aged 8-17 years old,in order to provide scientific evidence for preventing hypertension.Methods Seven thousand nine hundred and sixty-three primary and secondary students were sampled and standardized in blood pressure,height and weight.In accordance with the Working Group on Obesity in China(WGOC)recommendations,the subjects were divided into overweight,obesity and normal weight group,then the relationship between blood pressure and body mass index(BMI)and the prevalence of hypertension in different BMI groups were analyzed.Results 1.Compared with the USA and Shanghai city,the distribution of blood pressure in children and adolescents from the urban area of Guiyang city were different(Pa
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Objective To study the construction of the lentiviral-mediated RNA interference(RNAi) vector targering rat suppressors of cytokine signaling 3(SOCS3) gene.Methods Three target sequences were selected by on-line designer software on Ambion according to rat SOCS3 mRNA sequence(NM053565),the complementary DNA contained both sense and antisense oligonucleotides were designed and synthesized.After annealing,these double strands DNA were cloned to pRNA-Lenti-green fluorescent protein(GFP),which contained U6 promoter and GFP.The resulting Lentiviral vector containing SOCS3 shRNA was named pRNA-Lenti-SOCS3-GFP.After the rat glioma cells(C6)were transduced with the constructed 1entiviral vectors,real-time polymerase chain reaction was used to evaluate the level of SOCS3 expression(including siRNA1 group,siRNA2 group,siRNA3 group,vacuity group and siRNA-Negative group).The pRNA-Lenti-SOCS3-GFP and Lentivector Pakaging plasmid mix were cotransfected into 293T to package Lentivirus particles.Culture supematant was harvested,then the virus titer was determined by serial dilution assay.Results The SOCS3 mRNA sequence was successfully cloned to pRNA-Lenti-GFP,which was proved by PCR and DNA sequence.Compared with control group,the SOCS3 mRNA expressions were obviously suppressed in all 3 experimental groups,especially the expression rate in siRNA1 group was reduced by 80%.The Lentiviral particle titer was determined by serial dilution assay with 1.0?1010 TU?L-1.Conclusion The lentiviral-mediated RNAi vector of rat SOCS3 gene has been constructed successfully,this may provide a potential tool for studying and treating SOCS3-related diseases.
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Objective To study the changes of Na+K+-ATPase,Ca2+Mg2+-ATPase activities in erythrocyte membrane and blood viscosity in children with essential hypertension.Methods The activities of Na+K+-ATPase,Ca2+Mg2+-ATPase in erythrocyte membrane were determined by a colorimetric method.Blood viscosity was measured and analyzed with the statistic analysis SPSS 12.0 software in 50 children from Nov.2004 to Dec.2004 in the people's hospital of guizhou province and adolescents with essential hypertension.Thirty healthy children were collected as control group.Results The activities of Na+ K+-ATPase[(6.12?1.30)?molpi/(gHb?h)and(4.59?1.40)?molpi/(gHb?h)],Ca2+Mg2+-ATPase[(7.46?1.30)?molpi/(gHb?h)and(5.81?1.20)?molpi/(gHb?h)] were lower significantly in hypertension group than those in control group(Pa
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Objective To explore the hemorheological changes in children with essential hyepertension,and explore their clinical significance.Methods The hemorheological parameters,blood lipid,blood glucose in 37 children with essential hyepertension [19 boys and 18 girls,median age(12.49?3.20) years] were measured and analyzed.The parameters in 30 healthy children were measured as controls.SPSS 11.5 software was used to analyze the data.Results The whole blood viscosity under high,middle and low shear rate,plasma viscosity,hematocrit,rigidity index of erythrocyte,platelet aggregation M were significantly higher in hypertension group than those in healthy control group(Pa0.05).Conclusions There have been developed significant hemorheological changs in children with essential hypertension.The hemorheological changes might play an important role in pathogenesis of childhood hypertension.