The phenotype and genotype characteristics of 41 patients with steroid 5α-reductase type 2 deficiency / 中华内分泌代谢杂志

Objective To investigate the cliuical phenotype and the genotype of forty-one patients with steroid 5α-reductase type 2 deficiency.Methods The clinical data were collected including physical examination,medical history,laboratory test,as well as ultrasonic examination.Genomic DNA was extracted from peripheral blood leukocytes.Sanger sequencing and targeted gene captured next-generation sequencing were applied to detect the SRDSA2 gene mutation.Results All the patients are Han nationality and their ages ranged from 4 months to 11 years old.The karyotypes of 41 patients were 46,XY and all SRY genes were detected as positive.There were 26 (63％) patients manifested isolated micropenis,and the rest of fifteen patients were hypospadias associated with microphallus accounting for 37％.There were 39 patients who carried biallelic mutation.Two cases just identified one allele mutation.Sixteen gene mutation types were confirmed.Among them c.725A ＞ G (p.Tyr242Cys),c.694C ＞ G (p.His232Asp),and c.548-9T＞G are the novel gene types.The allele frequency of c.680G＞A (p.Arg227Gln) is 60％ (48/80).Conclusion The primary manifestations of patients with steroid 5α-reductase type 2 deficiency were micropenis or hypospadias accompanied with micropenis.c.680G＞A (p.Arg227Gln) is the predominantly mutation type of Chinese patient with steroid 5α-reductase type 2 deficiency.

Embarazo y adolescencia: comportamiento clínico-epidemiológico en el Policlínico Parraga: Arroyo Naranjo: 2005-2006 / Pregnancy and adolescents: epidemiologist and clinical features in Parraga Policlinic, Arroyo Naranjo: 2005-2006

Se presenta un estudio empírico, observacional, descriptivo y retrospectivo de 69 gestantes adolescentes captadas en el Policlínico Docente Párraga, Arroyo Naranjo, Ciudad de La Habana, en el período julio 2005 y julio del 2006. El propósito fue caracterizar el comportamiento social, epidemiológico y clínico del embarazo en las adolescentes. La información se obtuvo a través de la planilla de recolección de datos aplicada a las 69 pacientes con el auxilio del carné obstétrico y el Departamento de Estadística del Policlínico. Las variables estudiadas fueron: grupo etáreo, escolaridad, ocupación, estado conyugal, tipo de captación, planificación del embarazo, valoración nutricional, edad gestacional y características del parto, clasificación del peso y puntuación del Apgar. El grupo etáreo más afectado fue entre 17 y 19 años, predominaron las solteras y las uniones consensuales sobre las casadas. La Escuela Secundaria Básica constituyó el mayor nivel de escolaridad. El mayor número de los embarazos fue no deseado, coincidiendo con el mayor número de captaciones intermedias y tardías. Hubo predominio de las adolescentes normopeso, los casos de apgar bajo se relacionaron con los partos distócicos, mientras que en los recién nacidos bajo peso y riesgo por peso predominaron los partos pretérminos(AU)

It's an observational, descriptive and retrospective study of 69 pregnant adolescents captured in the Educational Policlinic Párraga of the Arroyo Naranjo Municipality Havana City, in the period understood between July 2005 and July of the 2006. The purpose was characterize the social behaviour, epidemiologist and clinical of the pregnancy in the adolescents. The information was gotten through the request data applied to 69 patients with the aid of the obstetric control and of the department of statistics of the Policlinic. The studied variables were: age group, educational level, occupation, and marital state, planning of the pregnancy, nutritional valuation, and gestational age to the childbirth, characteristic of the childbirth, classification of the weight and punctuation of the Apgar. The age group more affected was between 17 to 19 years, there prevailed the single and the consensual unions on the married. As for the educational level the secondary basic reached the biggest percent with a 59,4, and they constituted the predominant population. The biggest number of the pregnancies was not desired (47 cases), coinciding with the biggest number of intermediates and late citations. The adolescent's normal weight was the 60,8 percent, the cases of low Apgar were related distocics with the childbirths, it was also observed that in the newly born under weight and risk for weight prevailed the preterm births(AU)

Absence of PRKAR1A loss of heterozygosity in laser-captured microdissected pigmented nodular adrenocortical tissue from a patient with Carney complex caused by the novel nonsense mutation p. Y21X / Ausência da perda de heterozigose do PRKAR1A em células capturadas por microdissecção a laser de tecido de nódulo pigmentoso adrenocortical de um paciente com complexo de Carney causado por uma nova mutação nonsense

OBJECTIVE: Primary pigmented nodular adrenocortical disease (PPNAD) is the main endocrine manifestation of Carney complex, a multiple neoplasia syndrome caused by PRKAR1A gene mutations. The presence of PRKAR1A loss of heterozygosity (LOH) in adrenocortical tumorigenesis remains controversial. The aim of the present study is to investigate the presence of PRKAR1A LOH in adrenocortical cells in a patient with Carney complex. METHODS: The LOH was investigated using a PRKAR1A informative intragenic marker by GeneScan software analysis in DNA obtained from laser-captured microdissected cells of several adrenal nodules. Patients: A young adult male patient with Carney complex and his family were studied. RESULTS: A novel heterozygous mutation (p. Y21X) was identified at PRKAR1A in blood DNA of the male proband and his relatives. No PRKAR1A LOH was evidenced in the laser-captured microdissected cells from PPNAD tissue by different methodologies. CONCLUSION: We identified a new PRKAR1A nonsense mutation and in addition we did not evidence PRKAR1A LOH in laser-captured nodules cells, suggesting that adrenocortical tumorigenesis in PPNAD may occurs apart from the second hit.

OBJETIVO: A doença adrenocortical nodular pigmentosa primária (PPNAD) é uma das manifestações do complexo de Carney, uma neoplasia endócrina múltipla causada por mutações no PRKAR1A. A perda de heterozigose (LOH) do PRKAR1A na tumorigenese adrenal permanece controversa dada à possibilidade de contaminação com o tecido normal. Nosso objetivo foi investigar a presença de LOH no PRKAR1A a partir de células do nódulo adrenal de um paciente com complexo de Carney. MÉTODOS: A pesquisa da LOH do PRKAR1A foi realizada através do estudo de um marcador intragênico em DNA de células do nódulo adrenal microdissecadas a laser, evitando contaminação com o tecido normal. Pacientes: Um paciente com PPNAD e cinco familiares foram estudados. RESULTADOS: A nova mutação (p. Y21X) foi identificada no PRKAR1A sem evidência de LOH no tecido adrenal. CONCLUSÃO: Identificamos uma nova mutação no PRKAR1A e não evidenciamos LOH nas células dos nódulos adrenocorticais, sugerindo que a PPNAD possa ocorrer na ausência de um segundo evento molecular.

Eye Movement Recording using Computer Image Recognition Technique

PURPOSE: To describe that Computer image recognition technique is more accurately and less invasive than other methods, which using contact lens with a search probe or electrodes for record of eye movements. METHODS: A reference marker which was infrared was attached to the center of the forehead of the patients and the infrared light was illuminated to both eyes. Video image sequences were recorded using digital CCD camera and the captured image frames (640 X 480) were processed using a image analysis program. From which each image frame captured, pupil area was saperated using difference of brightness in pupil and iris. And then the pupil center was determined by calculating the center-of-mass of black pixels to meet the pupil threshold criteria. RESULTS: In a post-processing process, we could acquire horizontal and vertical eye position and velocity data. Inverse fast fourier transformation(FFT) and digital filtering algorithm were applied to filter out noise due to limited resolution of the CCD camera and sampling rates. Experimental result showed that the system could detect about 0.2mm positional location and 30Hz horizontal and vertical eye movements simultaneously. CONCLUSIONS: We could accurately measure and record two dimensional eye movements of patients (horizontal and vertical simultaneously) in real time with some head movements.

Analysis of the Gene Expression by Laser Captured Microdissection (I): Minimum Conditions Required for the RNA Extraction from Oocytes and Amplification for RT-PCR / 대한불임학회지

OBJECTIVE: Recently, microdissection of tissue sections has been used increasingly for the isolation of morphologically identified homogeneous cell populations, thus overcoming the obstacle of tissue complexity for the analysis cell-specific expression of macromolecules. The aim of the present study was to establish the minimal conditions required for the RNA extraction and amplification from the cells captured by the laser captured microdissection. METHODS: Mouse ovaries were fixed and cut into serial sections (7 micrometer thickness). Oocytes were captured by laser captured microdissection (LCM) method by using PixCell IITM system. The frozen sections were fixed in 70% ethanol and stained with hematoxylin and eosin, while the paraffin sections were stained with Multiple stain. Sections were dehydrated in graded alcohols followed by xylene and air-dried for 20 min prior to LCM. All reactions were performed in ribonuclease free solutions to prevent RNA degradation. After LCM, total RNA extraction from the captured oocytes was performed using the guanidinium isothiocyanate (GITC) solution, and subsequently evaluated by reverse transcriptase -polymerase chain reaction (RT-PCR) for glyceraldehyde-3-phosphate-dehydrogenase (GAPDH). RESULTS: With the frozen sections, detection of the GAPDH mRNA expression in the number of captured 25 oocytes were not repeatable, but the expression was always detectable from 50 oocytes. With 25 oocytes, at least 27 PCR cycles were required, whereas with 50 oocytes, 21 cycles were enough to detect GAPDH expression. Amount of the primary cDNA required for RT-PCR was reduced down to at least 0.25 microl with 50 oocytes, thus the resting 19.75 microl cDNA can be used for the testing other interested gene expression. Tissue-to-slide, tissue-to-tissue forces were very high in the paraffin sections, thus the greater number of cell procurement was required than the frozen sections. CONCLUSION: We have described a method for analyzing gene expression at the RNA level with the homogeneously microdissected cells from the small amount of tissues with complexity. We found that LCM coupled with RT-PCR could detect housekeeping gene expression in 50 oocytes captured. This technique can be easily applied for the study of gene expression with the small amount of tissues.