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<p><b>OBJECTIVE</b>To investigate the in vitro effects of different culture systems on hematopoietic differentiation ability of induced pluripotent stem (iPS) cells.</p><p><b>METHOD</b>Two culture systems including E8 and mTESR(freeder-free medium), and the classical ES culture medium were chosen for culture of iPS cells. The iPS cells maintaining in above mentioning culcure systems were co-cultured with OP9 cells(murine bone marrow stromal cells) in vitro to be induced to differentiate into hematopoietic stem/progenitor cells. Flow cytometry and real-time quantitative PCR were used to detect the expression of specific hematopoietic markers and the effects of different culture systems on the differentiation of iPS in vitro.</p><p><b>RESULT</b>iPS cultured in the 3 selected medium could be differentiated into hematopoietic stem cells. Efficiency of hematopoietic differentiation was up to 28.4% in classical ES culture system, which was significantly higher than that in E8 and mTESR system.</p><p><b>CONCLUSION</b>Under the co-culture with OP9, iPS can differentiate into hematopoietic stem/progenitor cells, which shows higher efficiency when iPS maintained in the ES medium.</p>
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<p><b>BACKGROUND</b>Fetal insulin hypothesis was proposed that the association between low birth weight and type 2 diabetes is principally genetically mediated. The aim of this study was to investigate whether common variants in genes CDKAL1, HHEX, ADCY5, SRR, PTPRD that predisposed to type 2 diabetes were also associated with reduced birthweight in Chinese Han population.</p><p><b>METHODS</b>Twelve single nucleotide polymorphisms (rs7756992/rs10946398 in CDKAL1, rs1111875 in HHEX, rs391300 in SRR, rs17584499 in PTPRD, rs1170806/rs9883204/rs4678017/rs9881942/rs7641344/rs6777397/rs6226243 in ADCY5) were genotyped in 1174 unrelated individuals born in Peking Union Medical College Hospital from 1921 to 1954 by TaqMan allelic discrimination assays, of which 645 had normal glucose tolerance, 181 had developed type 2 diabetes and 348 impaired glucose regulation. Associations of these 12 genetic variants with birthweight and glucose metabolism in later life were analyzed.</p><p><b>RESULTS</b>Birthweight was inversely associated with CDKAL1-rs10946398 (β = -41 g [95% confidence interval [CI]: -80, -3], P = 0.034), common variants both associated with increased risk of impaired glucose metabolism and decreased insulin secretion index later in life. After adjusting for sex, gestational weeks, parity and maternal age, the risk allele of CDKAL1-rs7756992 was associated with reduced birthweight (β = -36 g [95% CI: -72, -0.2], P = 0.048). The risk allele in SRR showed a trend toward a reduction of birthweight (P = 0.085).</p><p><b>CONCLUSIONS</b>This study identified the association between type 2 diabetes risk variants in CDKAL1 and birthweight in Chinese Han individuals, and the carrier of risk allele within SRR had the trend of reduced birthweight. This demonstrates that there is a clear overlap between the genetics of type 2 diabetes and fetal growth, which proposes that lower birth weight and type 2 diabetes may be two phenotypes of one genotype.</p>
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Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adenilil Ciclasas , Genética , Alelos , Pueblo Asiatico , Genética , Peso al Nacer , Genética , Quinasa 5 Dependiente de la Ciclina , Genética , Diabetes Mellitus Tipo 2 , Genética , Predisposición Genética a la Enfermedad , Genética , Proteínas de Homeodominio , Genética , Recién Nacido de Bajo Peso , Polimorfismo de Nucleótido Simple , Genética , Proteínas Tirosina Fosfatasas Clase 2 Similares a Receptores , Genética , Factores de Transcripción , Genética , ARNt MetiltransferasasRESUMEN
<p><b>OBJECTIVE</b>To reprogram amniotic fluid cells into pluripotent stem cells in order to create an optimal internal control model for directed cell differentiation.</p><p><b>METHODS</b>Human amniotic fluid-derived cells (hAFDCs) from heterozygotic twin fetuses were induced by retroviral vectors encoding Oct4, Sox2, c-Myc and Klf4. In vivo pluripotency, differentiation capacity and karyotype of hAFDCs induced pluripotent stem cells (hAFDCs-iPSCs) were determined.</p><p><b>RESULTS</b>hAFDC-iPSCs derived from heterozygotic twins have maintained self renewal, with expression of high pluripotency marker gene detected at both mRNA and protein levels. The cells have maintained their differentiation capacity both in vitro and vivo, and showed normal karyotypes after long-term culturing in vitro.</p><p><b>CONCLUSION</b>hAFDCs-iPSCs derived from heterozygotic twins have good consistency in terms of genetic background, and can provide a good internal control for directed differentiation of iPSCs, and may be used an ideal source for autologous cell replacement therapy in the later life of the fetus.</p>
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Femenino , Humanos , Embarazo , Líquido Amniótico , Biología Celular , Metabolismo , Diferenciación Celular , Genética , Línea Celular , Feto , Metabolismo , Heterocigoto , Células Madre Pluripotentes Inducidas , Biología Celular , Metabolismo , Cariotipo , Células Madre Pluripotentes , Biología Celular , Metabolismo , GemelosRESUMEN
<p><b>OBJECTIVE</b>To analyze de novo copy number variations (CNVs) in a Chinese family affected with autism spectrum disorders (ASD).</p><p><b>METHODS</b>Affymetrix Cytogenetics Whole Genome 2.7M Array assay was performed to identify potential CNVs in four members from the family.</p><p><b>RESULTS</b>A total of 89 de novo CNV regions were identified in the autistic siblings. The CNV regions in total have exceeded 1/1000 of the lengths of chromosomes 5, 11 and 14. In addition, de novo CNV regions were also identified at 3p26.1, 4q22.2, and 5p15.2, which encompassed 10 genes associated with nerve development including GRM7, GRID2 and CTNND2.</p><p><b>CONCLUSION</b>A number of nerve development associated genes were at the de novo CNV sites, which may provide new clues for genetic research of ASD. High-resolution array-comparative genomic hybridization is an effective method for detecting submicroscopic chromosomal imbalances.</p>
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Preescolar , Femenino , Humanos , Masculino , Trastornos Generalizados del Desarrollo Infantil , Genética , Hibridación Genómica Comparativa , Métodos , Variaciones en el Número de Copia de ADNRESUMEN
<p><b>OBJECTIVE</b>To identify potential mutations in patients featuring Becker muscular dystrophy (BMD) and to enhance the understanding of non-deletion/duplication mutations of the dystrophin gene causing BMD.</p><p><b>METHODS</b>Clinical data of two patients affected with BMD were collected. Potential mutations in the dystrophin gene were screened with multiplex ligation-dependent probe amplification assay (MLPA). Biopsied muscle samples were examined with HE staining, immnostaining with anti-dystrophin antibody, and electronic microscopy.</p><p><b>RESULTS</b>MLPA assay suggested that both cases were probably due to non-deletion/duplication mutations of the dystrophin gene. Light and electronic microcopy of skeletal muscle biopsies confirmed dystrophic changes in both patients. For patient A, immunostaining showed non-contiguous weak staining for most parts of sarcolemma. For patient B, immunostaining showed positive result with N-terminal anti-dystrophin antibody and negative result with C-terminal anti-dystrophin antibody.</p><p><b>CONCLUSION</b>For patients with mild phenotypes but without dystrophin gene deletion/duplication, muscle biopsy and immunochemistry are helpful for diagnosis and prognosis.</p>
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Adolescente , Adulto , Humanos , Masculino , Distrofina , Genética , Metabolismo , Músculo Esquelético , Patología , Distrofia Muscular de Duchenne , Genética , Metabolismo , Patología , Mutación , GenéticaRESUMEN
<p><b>BACKGROUND</b>Genome-wide association studies for type 2 diabetes mellitus (T2DM) identified FTO gene as a locus conferring increased risk for common obesity in many populations with European ancestry. However, the involvement of FTO gene in obesity or T2DM related metabolic traits has not been consistently established in Chinese populations. The objective of this study was to investigate the association of FTO genetic polymorphisms with metabolic syndrome (MetS) in Han Chinese.</p><p><b>METHODS</b>We tested 41 FTO single nucleotide polymorphisms (SNPs) for association between FTO and MetS-related traits. There were a total of 236 unrelated subjects (108 cases and 128 controls), grouped according to the International Diabetes Federation (IDF) criteria.</p><p><b>RESULTS</b>Of the 41 SNPs examined, only SNP rs8047395 exhibited statistical significance (P = 0.026) under a recessive model, after Bonferroni adjustment for multiple testing (OR 1.64, 95%CI 1.11-2.42; P = 0.014). The common distributions of this polymorphism among Chinese--with a minor allele frequency (MAF) of 36% in the control group versus 48% in the MetS group--greatly improved our test power in a relatively small sample size for an association study. Previously identified obesity- (or T2DM-) associated FTO SNPs were less common in Han Chinese and were not associated with MetS in this study. No significant associations were found between our FTO SNPs and any endophenotypes of MetS.</p><p><b>CONCLUSIONS</b>A more common risk-conferring variant of FTO for MetS was identified in Han Chinese. Our study substantiated that genetic variations in FTO locus are involved in the pathogenesis of MetS.</p>
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Femenino , Humanos , Masculino , Persona de Mediana Edad , Dioxigenasa FTO Dependiente de Alfa-Cetoglutarato , Pueblo Asiatico , Predisposición Genética a la Enfermedad , Genética , Variación Genética , Genética , Genotipo , Haplotipos , Genética , Síndrome Metabólico , Genética , Polimorfismo de Nucleótido Simple , Genética , Proteínas , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To compare the development of abnormal pronuclear zygotes after intracytoplasmic sperm injection (ICSI) and analyze their genetic polymorphism.</p><p><b>METHODS</b>Four hundred and ninety three abnormal pronuclear zygotes after ICSI were divided into three groups based on the number of pronuclei: 347 nonpronuclear oocytes, 71 monopronuclear zygotes and 75 multipronuclear zygotes. All of them were cultured in the medium of Vitrolife G5 series(TM). Sixteen short tandem repeats (STR) of seven blastocysts were then analyzed by ABI3100.</p><p><b>RESULTS</b>The cleavage rate of nonpronuclear group (25.4%) was lower than that of the others (P<0.01), the proportion of blocked embryos in nonpronuclear group (48.9%) was significantly higher than that of the others (P<0.05), but the blastocyst rate showed no significant difference in three groups (P>0.05). The genetic polymorphism of the 16 STRs showed that the blastocysts from the nonpronuclear and multipronuclear were diploid, and one of the blastocysts from nonpronuclear oocyte was Y-bearing.</p><p><b>CONCLUSION</b>The zygotes with abnormal pronuclei after ICSI might have development potential, and the blastocysts from nonpronuclear oocytes and multipronuclear zygotes could be diploid.</p>
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Femenino , Humanos , Masculino , Blastocisto , Fisiología , Núcleo Celular , Fisiología , Desarrollo Embrionario , Genética , Fisiología , Fertilización In Vitro , Oocitos , Fisiología , Inyecciones de Esperma Intracitoplasmáticas , Secuencias Repetidas en Tándem , Cigoto , FisiologíaRESUMEN
<p><b>OBJECTIVE</b>Discussion of the relationship between Mycoplasma and chlamydia infection and lesions in the cervical tissue in high-risk HPV-positive infertile patients with cervical.</p><p><b>METHODS</b>HPV-negative patients with cervical as the control, retrospective analysis the relationship of Mycoplasma hominis and chlamydia infection, cervical histological graded, and inflammation graded.</p><p><b>RESULTS</b>The rate of HPV infection in mycoplasma-positive and those with negative mycoplasma has significant difference (P < 0.01), The rate of HPV infection in chlamydia-positive and those with negative chlamydia has no significant difference (P > 0.05). CIN and the incidence of cervical erosion and CIN grade were higher in HPV-positive than HPV-negative group (P < 0.01). The cervical erosion of HPV-positive was no difference in the degree (P > 0.05). Compared with the simple HPV-positive group, CIN and the incidence of severe cervical erosion in mixed infection of Mycoplasma was no difference (P > 0.05).</p><p><b>CONCLUSION</b>Mycoplasma infection increases the rate of high risk HPV infection, high-risk HPV infection increased cervical pathological damage, Mycoplasma infection might be the factor of persistent infection with high risk HPV, the degree of cervical pathological is the factor of cervical infertility which can not be ignored.</p>
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Adulto , Femenino , Humanos , Adulto Joven , Alphapapillomavirus , Genética , Cuello del Útero , Microbiología , Patología , Virología , Chlamydia , Infecciones por Chlamydia , Microbiología , Patología , Virología , Infertilidad Femenina , Microbiología , Patología , Virología , Mycoplasma , Infecciones por Mycoplasma , Microbiología , Patología , Virología , Infecciones por Papillomavirus , Microbiología , Patología , Virología , Estudios Retrospectivos , Factores de RiesgoRESUMEN
<p><b>OBJECTIVE</b>Explore the optimal treatment of infertility patients infected with different types of human papillomavirus (HPV).</p><p><b>METHODS</b>According to cervical pathology, cervical status and the procreate desire of the infertility patients, the 144 clinic cases of high-risk human papillomavirus infected infertile patients were divided into two gruoups: group with treatment and without treatment. Real-time quantitative fluorescent PCR (RT-PCR) has been employed, follow-up time is 6 months, to detect the HPV-DNA in the crevical exfoliated cells, to observe the negative conversion rate and pregnancy rate, and compare analyzed.</p><p><b>RESULTS</b>(1) In high-risk HPV infectors, the negative conversion rate of treatment group (56.67%) is higher than those in non-treatment group (50.00%); (2) The pregnancy rate of secondary high-risk HPV non-treatment group (50.00%) is higher than the treatment group. The pregnancy rate of primary high-risk HPV treatment group (31.67%) is higher than the non-treatment group (4.00%). (3) Negative conversion rate increases accordingly, on primary high-risk HPV infected groups with Leep, with single drug and with Leep combined with drug therapy. (4) The negative conversion rate and the pregnancy rate of primary high-risk HPV infected groups with surgical therapy is higher than the groups with drug therapy. Surgical + Drugs is better in the two surgical therapies.</p><p><b>CONCLUSION</b>Infertile patients should be routinely screened for cervical HPV. The primary high-risk cervical HPV infection is the etiology of infertility. Preferably, patients with primary high-risk HPV infection in cervical lesions is treated with Leep combined drugs.</p>
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Adulto , Femenino , Humanos , Embarazo , ADN Viral , Quimioterapia , Infertilidad , Virología , Papillomaviridae , Genética , Infecciones por Papillomavirus , Quimioterapia , Epidemiología , Terapéutica , Preparaciones Farmacéuticas , Reacción en Cadena de la Polimerasa , Índice de Embarazo , Factores de Riesgo , Experimentación Humana TerapéuticaRESUMEN
<p><b>OBJECTIVE</b>To investigate the effect of sperm acrosin activity on the IVF-ET outcome.</p><p><b>METHODS</b>We analyzed sperm parameters, morphology and acrosin activity for 909 infertile husbands by computer-assisted self-assessment (CASA), modified Papanicolaou staining and N-alpha-benzoyl-DL-arginine-p-nitroanilide (BAPNA), respectively, and detected the rates of fertilization, cleavage, quality embryos, embryo cryopreservation, implantation, clinical pregnancy and abortion. The wives were identified as normal or with mere oviduct problems.</p><p><b>RESULTS</b>The rate of normal sperm morphology and sperm motility, vitality, rapid progressive velocity and concentration were significantly lower in the abnormal acrosin activity group than in the normal one (P < 0.01). Significant positive correlations were observed between acrosin activity and the above-mentioned semen parameters (P < 0.01). There were no significant differences in the number of retrieved eggs, the rates of cleavage, quality embryos, embryo cryopreservation, non-embryo transfer cycles and miscarriages, and the number of transferred embryos between the two groups (P > 0.05). The fertilization rate, the percentage of transfer cycles with only 1 embryo and the rate of implantation and clinical pregnancy were notably higher in the normal acrosin activity group than in the abnormal one (P < 0.01).</p><p><b>CONCLUSION</b>Sperm acrosin activity is closely related with semen parameters, and it helps to predict the sperm fertilizing capacity and IVF-ET outcome.</p>
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Adulto , Femenino , Humanos , Masculino , Embarazo , Acrosina , Metabolismo , Transferencia de Embrión , Fertilización In Vitro , Infertilidad Masculina , Índice de Embarazo , Análisis de Semen , EspermatozoidesRESUMEN
<p><b>OBJECTIVE</b>To investigate the clinical and lab features of sibling brother and sister both with Duchenne muscular dystrophy (DMD).</p><p><b>METHODS</b>We conducted comprehensive clinical and lab investigations including the test of serum enzymes, electromyography (EMG), electrocardiography, color Doppler echocardiography, HE staining of skeletal muscles, immunohistochemical study of dystrophin and utrophin, multiple ligation probe amplification (MLPA) on exon 1-79 of dystrophin gene, and short tandem repeat-poly- merase chain reaction of CA repeats located in dystrophin gene.</p><p><b>RESULTS</b>These two patients were confirmed to suffer from DMD. They were characterized by typical features of DMD including typical clinical manifestations, increased serum enzymes, EMG presenting myogenic impairment, HE staining presentation belonging to DMD, negative dystrophin in brother, and inconstantly positive on the sarcolemma of sister. Furthermore, no deletion or duplication was found in the 1-79 exons of dystrophin gene. The suffering brother and sister carried the same maternal X chromosome.</p><p><b>CONCLUSIONS</b>Carriers of DMD gene show typical clinical and laboratory manifestations of DMD. Comprehensive examinations should be performed for such carriers.</p>
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Femenino , Humanos , Masculino , Distrofina , Genética , Ligamiento Genético , Heterocigoto , Distrofia Muscular de Duchenne , Genética , Metabolismo , HermanosRESUMEN
<p><b>OBJECTIVE</b>To compare the effectiveness of using multiple ligation probe amplification (MLPA) and denaturing high-performance liquid chromatography (DHPLC) in screening the exon deletions and duplications of the DMD gene.</p><p><b>METHODS</b>MLPA technique was applied to detect exon deletions and duplications previously confirmed by denaturing high-performance liquid chromatography (DHPLC).</p><p><b>RESULTS</b>From October 2004 to October 2005, 22 unrelated DMD probands and their possible female relatives with clinical diagnosis with dystrophinopathy at our hospital entered this study. Both DHPLC and MPLA detected DMD gene depletion in 11 probands and DMD duplications in 3 probands. MLPA detected deletions and duplications in 2 probands, which were not detected by DHPLC. MLPA also successfully identified the carriage status of the potential female carriers of the probands.</p><p><b>CONCLUSION</b>Compared with DHPLC and traditional PCR techniques, MLPA is a superior tool to analyze the deletions and duplications in affected males as well as in the identification of the carriage status of potential females carriers.</p>
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Femenino , Humanos , Masculino , Cromatografía Líquida de Alta Presión , Eliminación de Gen , Duplicación de Gen , Predisposición Genética a la Enfermedad , Distrofia Muscular de Duchenne , Genética , Mutación , Técnicas de Amplificación de Ácido Nucleico , MétodosRESUMEN
<p><b>OBJECTIVE</b>To detect genomic deletion and duplication mutations in the dystrophin gene of the Duchenne muscular dystrophy (DMD) patients and their potential female carriers.</p><p><b>METHODS</b>Genomic deletions and duplications of the DMD gene in 32 affected males and 27 potential female carriers were screened by mutiplex ligation-dependent probe amplification (MLPA).</p><p><b>RESULTS</b>Of the 32 investigated affected males, 24 were detected to have deletions of one or more exons of the DMD gene, 1 patient had a duplication from exon 5 to 55, 1 patient had a nonsense point mutation (R768X) in exon 19, the other 6 affected males were predicted to have possible disease-causing point mutations. MLPA analysis showed a DMD deletion or duplication in 18 female relatives, and the female carriers had the same deletion or duplication as their probands, respectively.</p><p><b>CONCLUSION</b>MLPA analysis is proven to be an efficient tool for identification of both affected males and female carriers of DMD rearrangements in cases in which the disease-causing mutation in the affected male was not known. It could provide useful information for the genetic counseling of the family involved.</p>
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Femenino , Humanos , Masculino , Codón sin Sentido , Análisis Mutacional de ADN , Métodos , Distrofina , Genética , Duplicación de Gen , Predisposición Genética a la Enfermedad , Genética , Genotipo , Heterocigoto , Distrofia Muscular de Duchenne , Genética , Mutación Puntual , Eliminación de SecuenciaRESUMEN
<p><b>OBJECTIVE</b>To establish a protocol for culturing of human embryonic stem cells (HUES4) without any animal-derived feeder cells and to investigate the karyotype stabilities of HUES4 cells after long-term cultivation.</p><p><b>METHODS</b>HUES4 cells were cultured on mitomycin C treated MEFs or human foreskin fibroblast feeder cells. The pluripotency of the ES cells was analyzed by immunocytochemistry staining to detect the expression of pluripotent marker, karyotype of the ES cells at passage 27, 34, 41, 44 and short tandem repeat (STR) at passage 27 were analyzed.</p><p><b>RESULTS</b>The HUES4 cells cultured on human feeder cells were positive for alkaline phosphatase activity, SSEA-4, TRA-1-60 and TRA-1-81 staining, but negative for SSEA-1. Analysis of karyotype at different passages suggested an abnormal karyotype 46, XY, t(9;15)(q22;q26) mosaicism occurred in HUES4, and the ratios of abnormal increased with passage.</p><p><b>CONCLUSION</b>HUES4 could be cultured without animal-derived feeder cells and the incidence of abnormal karyotype might be increased with long-term culture.</p>
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Humanos , Masculino , Técnicas de Cultivo de Célula , Métodos , Línea Celular , Proliferación Celular , Células Madre Embrionarias , Biología Celular , Metabolismo , Fibroblastos , Biología Celular , Metabolismo , Prepucio , Biología Celular , Inmunohistoquímica , Cariotipificación , Métodos , Repeticiones de Microsatélite , Genética , Polimorfismo GenéticoRESUMEN
<p><b>OBJECTIVE</b>High risk human papilomavirus (HPV) infection is often related to cervical cancer. This study investigated the infection of high risk HPV in cervical epithelia among infertile patients. Relative quantification and absolute quantification were applied for determination of "real" HPV viral load in the clinical setting.</p><p><b>METHODS</b>Adopting multi-channels real time PCR to genotype and quantify eight high risk HPV (HPV16, 18, 45, 31; intermediate risk types: HPV33, 52, 58, 67) DNA in cervical epithelia of the 130 infertile patients and the 150 controls. This study applied housekeeping gene (beta-globin) for the DNA quantification on secretions samples for clinical diagnosis.</p><p><b>RESULTS</b>The infection rate of the infertility group was 25.38 percent (33/130) and that of the control group was 11.33 percent (17/150), the difference was statistically significant. Among the 33 positive cases in the infertility group, 24 cases showed a viral load no less than 106; in 9 of them, the viral load was less than 106. Among the 17 positive cases in the control group, 4 cases had a viral load no less than 106; in 13 of them, the viral load was less than 106. There is a statistically significant difference in viral load between the infertility group and the control group.</p><p><b>CONCLUSION</b>The HPV infection rate of the infertility group was higher than that of the control group.</p>
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Adulto , Femenino , Humanos , Adulto Joven , Alphapapillomavirus , Genética , Infertilidad Femenina , Virología , Infecciones por Papillomavirus , Virología , Frotis Vaginal , Carga ViralRESUMEN
<p><b>OBJECTIVE</b>To detect the disease-causing point mutations in the dystrophin gene of Duchenne muscular dystrophy (DMD) patients.</p><p><b>METHODS</b>The approach of denaturing high performance liquid chromatography (DHPLC) coupling with sequencing was used to screen the point mutations of 79 exons and the untranslated regions of dystrophin gene without large deletions/duplications, which was in 6 unrelated DMD probands from 6 DMD families.</p><p><b>RESULTS</b>Five disease-causing mutations, 697-698insGT, C616T, G1255T, C4279T, and C2302T, were ides created the new stop codons in downstream sites of mutations, respectively. In addition to the disease-causing point mutations, a point mutation T5586+61A in intron 39 was also found at patient 3, and a missense mutation A694T in exon 8 was detected at patient 5. Four point mutations, C2168+13T, 5740-13dupG, G5234A and C5280T, were also detected at patient 6 whose causative point mutation was unavailable. Seven point mutations have not been reported previously. Bi-directional PCR amplification of specific alleles (Bi-PASA) method was established to distinguish the haplotypes of heterozygote or homozygote in a single PCR reaction.</p><p><b>CONCLUSION</b>Via automated DHPLC screening or detecting the subexonic mutations in dystrophin gene is feasible to clinical laboratories, and also is a superior method in terms of sensitivity and efficiency.</p>
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Humanos , Masculino , Secuencia de Bases , Cromatografía Líquida de Alta Presión , Análisis Mutacional de ADN , Distrofina , Genética , Duplicación de Gen , Distrofia Muscular de Duchenne , Genética , Mutación Puntual , Reacción en Cadena de la Polimerasa , Eliminación de SecuenciaRESUMEN
<p><b>OBJECTIVE</b>To investigate the effects of Tianzhi Keli (TZ) on acetylcholine (ACh) and catecholamine levels in striatum of rats with neuromitochondrial impairment, and try to find out the neuroprotective mechanism of TZ.</p><p><b>METHOD</b>The microdialysis and high performance liquid chromatography (HPLC)-post column Immobilized enzyme reactor (IMER)-electrochemical detection (ED) were used to establish a model of mitochondrial energy metabolism impairment which induced by perfusion with sodium azide (NaN3), and measure continuously the effects of TZ on extracellular ACh, choline (Ch) and catecholamine of model rats.</p><p><b>RESULT</b>After perfusion with NaN3, ACh, noradrenalin (NE), adrenaline (E), dopamine (DA), 3,4-Dihydroxyphenyl-aletic (DOPAC), and homovanillic acid (HVA) levels were decreased obviously (P < 0.05-0.01), while Ch level was increased distinctly (P < 0.01). Transmitters levels were recovered individually after stop the perfusion with NaN3. TZ can postpone the decrease of ACh and advance the recover of Ch. The effect of TZ coupled with duxil on increasing ACh level is more obviously than effect of TZ or duxil. TZ is also showing a tendency to postpone the decrease of catecholamine and advance its recovery. TZ coupled with duxil can advance the recovery of DOPAC and adjust the metabolic abnormity positively.</p><p><b>CONCLUSION</b>TZ has effect on protecting impairment of choline neurosystem, which induced by damage of mitochondrion and abnormity of energy metabolism; coupled with duxil have synergistic action. TZ also has tendency to protect the impairment of epinephrine and dopamine neurosystem.</p>
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Animales , Masculino , Ratas , Ácido 3,4-Dihidroxifenilacético , Metabolismo , Acetilcolina , Metabolismo , Catecolaminas , Metabolismo , Cuerpo Estriado , Metabolismo , Dopamina , Metabolismo , Combinación de Medicamentos , Medicamentos Herbarios Chinos , Farmacología , Espacio Extracelular , Metabolismo , Gastrodia , Química , Microdiálisis , Enfermedades Mitocondriales , Metabolismo , Norepinefrina , Metabolismo , Plantas Medicinales , Química , Ratas Sprague-Dawley , Azida Sódica , Uncaria , QuímicaRESUMEN
<p><b>OBJECTIVE</b>To investigate the effect of Tianma Gouteng recipe (TGR) on interfering left ventricular (LV) and aortic hypertrophy and tissue angiotensin II (Ang II) in rats with renovascular hypertension.</p><p><b>METHOD</b>The animal model of renovascular hypertension was used in this experiment. Hypertensive rats were randomly allocated into model group, Enalapril group and TGR group, and the drugs were used for 6 weeks continuously. During this period, the blood pressure of rats was measured every two weeks. After rats were sacrificed, the wet weight, tissue Ang II level of LV and aorta, and the cardiac index were measured.</p><p><b>RESULT</b>One week after renovascular stenosis, the systolic blood pressure (SPS) of model group was increased by 37.4 mmHg, and 7 weeks after stenosis, the LV and aortic hypertrophy was obvious increased, meanwhile, tissue Ang II of LV and aorta was raised markedly (P < 0.01). Contrasting with the model group, blood pressure was reduced and the morphological index was improved in Enalapril group respectively (P < 0.05 or P < 0.01); the wet weight of LV and aorta were reduced, the morphological index was improved, the rise of Ang II in tissue was suppressed, in TGR group significantly.</p><p><b>CONCLUSION</b>TGR can attenuate myocardial and aorta hypertrophy induced by renovascular hypertension, and suppress the rise of Ang II in tissue significantly. This suggests that TGR has the effects on interfering LV and aortic hypertrophy by an independent-antihypertensive way.</p>
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Animales , Masculino , Ratas , Angiotensina II , Metabolismo , Antihipertensivos , Farmacología , Aorta , Metabolismo , Patología , Combinación de Medicamentos , Medicamentos Herbarios Chinos , Farmacología , Enalapril , Farmacología , Gastrodia , Química , Hipertensión Renovascular , Hipertrofia Ventricular Izquierda , Metabolismo , Patología , Plantas Medicinales , Química , Distribución Aleatoria , Ratas Wistar , Uncaria , QuímicaRESUMEN
The effects of Gastrodia elata on preventing decrepitude and advancing memory are closely associated with its neuroprotective activity. Previous researches proved that G. elata, its active components and preparations played a neuroprotective role by affecting the excitotoxicity, nitric monoxide (NO) system, neuroglia, biomembrane, oxidative neurotoxicity, apoptosis et al. Recent researches also suggest that reducing energy metabolism impairment, anti-inflammatory and immune modulating function may be new research targets of neuroprotective mechanism of G. elata.
Asunto(s)
Animales , Humanos , Antioxidantes , Farmacología , Apoptosis , Calcio , Metabolismo , Medicamentos Herbarios Chinos , Farmacología , Aminoácidos Excitadores , Gastrodia , Química , Fármacos Neuroprotectores , Farmacología , Óxido Nítrico , Metabolismo , Óxido Nítrico Sintasa , Metabolismo , Plantas Medicinales , QuímicaRESUMEN
<p><b>OBJECTIVE</b>To observe the effects of Tianma Gouteng Fang (TGF) on the transmitter amino acids in the hippocampus extracellular liquids in freely moving rats subjected to incomplete brain ischemia.</p><p><b>METHOD</b>Hippocampus extracellular liquids was collected continuously by the microdialysis sampling technology in freely moving rats during pre-ischemia, incomplete ischemia and reperfusion periods induced by the occlusion and loose of both common carotid arteries. Each dialysate sample was assayed for GABA, Tau, Glu, Cys and Arg with HPLC-electrochemical detector.</p><p><b>RESULT</b>TGF increased the concentrations of GABA and Tau in the extracellular liquids of rat hippocampus. Compared with the model group, the concentration of Glu in the middle and large dosage groups of TGF, during the 120 min of ischemia, reduced by 38.64% and 31.35%, Tau increased by 13.99% and 12.86%, GABA advanced 25.89% and 33.99%, Cys decreased by 40.93% and 42.08%, Arg raised to 116.95% and 108.96%, respectively. After 120 min of reperfusion, the concentration of Glu decreased by 14.55% and 11.48%, Tau increased by 16.13% and 14.03%, GABA increased by 24.41% and 26.22%, respectively.</p><p><b>CONCLUSION</b>TGF can increase the concentration of inhibitory amino acids in hippocampus extracellular liquids of rats and inhibit the excessive release of excitatory amino acids and raise the concentration of the inhibitory amino acids and Arg during the ischemia-reperfusion periods. Therefore, TGF can play the neuroprotective role.</p>