Application of pyroptosis score in the treatment and prognosis evaluation of gastric cancer. / 细胞焦亡评分在胃癌治疗和预后评估中的应用.

Pyroptosis is a kind of proinflammatory programmed cell death mediated by inflammasome. It affects the occurrence and development of gastric cancer through different ways, showing dual effects. On the one hand, inflammasome-mediated inflammatory response is highly likely to participate in the formation and development of early tumors; on the other hand, drugs can inhibit the deterioration process of tumor proliferation, invasion and metastasis through activating the pathways of inflammasome and pyroptosis. Recently, many agents based on pyroptosis have been found to inhibit gastric cancer by promoting the secondary pyroptosis pathway, regulating NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome and inhibiting caspase-1. The establishment of cell pyrodeath models can predict the prognosis of gastric cancer patients. Most of the models show that gastric cancer patients with high pyroptosis level have better prognosis and longer overall survival. Pyroptosis scores can also be used to predict the response of gastric cancer patients to immunotherapy and to screen potential anti-gastric cancer drugs. Therefore, in-depth understanding of the potential mechanism of pyroptosis affecting the progression of gastric cancer and the role of pyroptosis score in the treatment and prognosis assessment of gastric cancer will be helpful to find a new and effective method for the treatment of gastric cancer.

Aerobic training method in throwing application / Aplicação do método de treinamento aeróbico em arremessos / Aplicación del método de entrenamiento aeróbico en los lanzamientos

ABSTRACT Introduction Strength training is a vital training modality for sports. Upper body strength training is critical for throwing. Aerobic activity can effectively develop upper body strength in throwing athletes. Objective Analyze the effect of aerobic training on upper body strength in throwing athletes. Methods Several ball pitchers were selected as research volunteers. They were randomly divided into control and experimental groups. The study adopted the method of establishing statistics to analyze the strength and performance of throwing athletes before and after aerobic training. This work also analyzed the relationship between aerobics and throwing ability. Results There were significant differences in the upper limb strength of throwing athletes after aerobic intervention (P<0.05). The strength of the experimental group was significantly improved (P<0.05). Conclusion The aerobic intervention method is an effective way to improve upper limb strength in throwing athletes. It is recommended that athletes apply upper limb strength training to their daily training. The aerobic training method is a safe and effective choice. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMO Introdução O treinamento de força é uma modalidade de treinamento vital para a prática de esportes. O treinamento de força da parte superior do corpo é fundamental para o arremesso. A atividade aeróbica pode efetivamente desenvolver a força nos membros superiores de atletas arremessadores. Objetivo Analisar o efeito do treinamento aeróbico sobre a força dos membros superiores de atletas arremessadores. Métodos Selecionou-se vários arremessadores de bolas como voluntários de pesquisa. Estes foram divididos aleatoriamente em grupo de controle e experimental. O trabalho adotou o método do estabelecimento de estatísticas para analisar a força e o desempenho dos atletas arremessadores antes e depois do treinamento de aeróbica. Paralelamente, este trabalho também analisou a relação entre a aeróbica e a capacidade de arremesso. Resultados Houve diferenças significativas na força dos membros superiores dos atletas arremessadores após a intervenção aeróbica (P<0,05). A força do grupo experimental foi significativamente aperfeiçoada (P<0,05). Conclusão O método de intervenção aeróbica é uma maneira eficaz de melhorar a força nos membros superiores de atletas arremessadores. Recomenda-se aos atletas aplicar o treinamento da força dos membros superiores ao treinamento diário. O método de treinamento de aeróbica é uma escolha segura e eficaz. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

RESUMEN Introducción El entrenamiento de fuerza es una modalidad de entrenamiento vital para la práctica deportiva. El entrenamiento de la fuerza de la parte superior del cuerpo es fundamental para el lanzamiento. La actividad aeróbica puede desarrollar eficazmente la fuerza en los miembros superiores de los atletas de lanzamiento. Objetivo Analizar el efecto del entrenamiento aeróbico en la fuerza de la parte superior del cuerpo de los atletas de lanzamiento. Métodos Se seleccionaron varios lanzadores de pelota como voluntarios para la investigación. Se dividieron aleatoriamente en grupos de control y experimental. El trabajo adoptó el método de establecer estadísticas para analizar la fuerza y el rendimiento de los atletas de lanzamiento antes y después del entrenamiento aeróbico. Paralelamente, este trabajo también analizó la relación entre el aeróbic y la capacidad de lanzamiento. Resultados Hubo diferencias significativas en la fuerza de las extremidades superiores de los atletas de lanzamiento después de la intervención aeróbica (P<0,05). La fuerza del grupo experimental mejoró significativamente (P<0,05). Conclusión El método de intervención aeróbica es una forma eficaz de mejorar la fuerza de las extremidades superiores en los atletas de lanzamiento. Se recomienda que los deportistas apliquen el entrenamiento de fuerza de las extremidades superiores en su entrenamiento diario. El método de entrenamiento aeróbico es una opción segura y eficaz. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

Impact of plyometric training on the lower limb strength of gymnasts / Impactos do treino pliométrico na força dos membros inferiores de ginastas / Impacto del entrenamiento pliométrico en la fuerza de los miembros inferiores de gimnastas

ABSTRACT Introduction: Plyometric training consists of a compound of stretching followed by immediate contraction, favoring the elastic properties of the soft tissues and aiming to promote a higher explosive force in the athletes. Objective: Explore the impact of plyometrics on aerobic gymnastics practitioners' explosive force in the lower limbs. Methods: In this experiment, a total of 16 aerobic gymnastics athletes were selected and divided into two groups: the control group and the experimental group. The control group remained with their usual training, while a sport-specific plyometric protocol was added to the experimental group. Results: Composite plyometric training can improve the short-distance running ability of aerobic gymnastics athletes; the effect of running 5m in the start and 10m in the start was statistically evidenced. However, the 20m run had similar results. Conclusion: The explosive power of the lower limbs in different activities is related to the specific training of aerobic gymnastics athletes focused on the muscles corresponding to the activity, showing a significant positive correlation. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMO Introdução: O treinamento pliométrico consiste num composto de alongamento seguido de imediata contração, favorecendo as propriedades elásticas dos tecidos moles visando promover uma maior força de explosão nos atletas. Objetivo: Explorar os impactos da pliometria sobre a força explosiva nos membros inferiores dos praticantes de ginástica aeróbica. Métodos: Neste experimento, um total de 16 atletas em ginástica aeróbica foram selecionados e divididos em dois grupos: o grupo de controle e o grupo experimental. O grupo controle permaneceu com seu treinamento usual enquanto ao grupo experimental foi acrescido um protocolo pliométrico específico para o esporte. Resultados: O treinamento composto de pliometria pode melhorar a capacidade de corrida de curta distância de atletas de ginástica aeróbica, o efeito de correr 5m em largada e 10m em largada foi estatisticamente evidenciado, porém a corrida de 20m não teve resultados diferentes. Conclusão: O poder explosivo dos membros inferiores em diferentes atividades está relacionado ao treinamento específico dos atletas de ginástica aeróbica focado nos músculos correspondentes à atividade, evidenciando uma correlação positiva significativa. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

RESUMEN Introducción: El entrenamiento pliométrico consiste en un compuesto de estiramientos seguido de una contracción inmediata, favoreciendo las propiedades elásticas de los tejidos blandos con el objetivo de promover una mayor fuerza explosiva en los deportistas. Objetivo: Explorar los impactos del entrenamiento pliométrico sobre la fuerza explosiva en los miembros inferiores de los practicantes de gimnasia aeróbica. Métodos: En este experimento, se seleccionaron un total de 16 atletas de gimnasia aeróbica y se dividieron en dos grupos: el grupo de control y el grupo experimental. El grupo de control siguió con su entrenamiento habitual mientras que al grupo experimental se le añadió un protocolo pliométrico específico para el deporte. Resultados: El entrenamiento compuesto de pliometría puede mejorar la capacidad de correr distancias cortas de los atletas de gimnasia aeróbica, se evidenció estadísticamente el efecto de correr 5m en salida y 10m en salida, sin embargo, la carrera de 20m no tuvo resultados diferentes. Conclusión: La potencia explosiva de los miembros inferiores en diferentes actividades está relacionada con el entrenamiento específico de los atletas de gimnasia aeróbica centrado en los músculos correspondientes a la actividad, evidenciando una correlación positiva significativa. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

Adaptation of cardiorespiratory endurance in obese people by aerobic exercise / Adaptação da resistência cardiorrespiratória de pessoas obesas por exercício aeróbico / Adaptación de la resistencia cardiorrespiratoria personas obesas mediante ejercicio aeróbico

ABSTRACT Introduction Obesity is a critical pathogenic factor of hypertension and hyperlipidemia in metabolic syndrome and an independent risk factor of cardiovascular disease leading to low cardiorespiratory endurance. Objective Explore the changes in cardiorespiratory endurance adaptation on obese subjects caused by aerobic exercise. Methods A cardiorespiratory exercise model was proposed for obese people under different optimizations based on critical variables and cluster analysis. This model analyzes the relationship between exercise and cardiorespiratory endurance in obese people, extracts the cardiorespiratory endurance index characteristics of obese people under different exercise levels, and clusters their different index characteristics. Results The difference between the heart rate of the proposed model and the actual heart rate is 0.11, the difference between the heart rate of the model and the actual heart rate is 4.28, and the difference between the heart rate of the model and the actual heart rate is 2.84, and the accuracy of the proposed model is the highest. Conclusion The model proposed in this paper can accurately analyze the effects of different aerobic exercise frequencies on cardiorespiratory endurance. Evidence Level II; Therapeutic Studies - Investigating the result.

RESUMO Introdução A obesidade não é apenas um fator patogênico chave da hipertensão e da hiperlipidemia na síndrome metabólica, mas também um fator de risco independente de doença cardiovascular que leva a baixa resistência cardiorrespiratória. Objetivo Explorar as alterações na adaptação da resistência cardiorrespiratória em obesos causadas pelo exercício aeróbico. Métodos Foi proposto um modelo de exercício cardiorrespiratório para pessoas obesas sob diferentes otimizações com base em variáveis-chave e análise de agrupamento. O modelo analisa a relação entre o exercício e a resistência cardiorrespiratória de pessoas obesas, extrai as características do índice de resistência cardiorrespiratória de pessoas obesas sob diferentes níveis de exercício e agrupa suas diferentes características de índice. Resultados A diferença entre a frequência cardíaca do modelo proposto e a frequência cardíaca real é de 0,11, a diferença entre a frequência cardíaca do modelo e a frequência cardíaca real é de 4,28, e a diferença entre a frequência cardíaca do modelo e a frequência cardíaca real é de 2,84, e a precisão do modelo proposto é a mais alta. Conclusão O modelo proposto neste trabalho pode analisar com precisão os efeitos de diferentes frequências de exercícios aeróbicos sobre a resistência cardiorrespiratória. Nível de evidência II; Estudos Terapêuticos - Investigação de Resultados.

RESUMEN Introducción La obesidad no sólo es un factor patogénico clave de la hipertensión y la hiperlipidemia en el síndrome metabólico, sino también un factor de riesgo independiente de enfermedad cardiovascular que conduce a una baja resistencia cardiorrespiratoria. Objetivo Explorar los cambios en la adaptación de la resistencia cardiorrespiratoria en sujetos obesos provocados por el ejercicio aeróbico. Métodos Se propuso un modelo de ejercicio cardiorrespiratorio para personas obesas bajo diferentes optimizaciones basadas en variables-clave y en el análisis de grupos. El modelo analiza la relación entre el ejercicio y la resistencia cardiorrespiratoria en las personas obesas, extrae las características del índice de resistencia cardiorrespiratoria de las personas obesas bajo diferentes niveles de ejercicio y agrupa sus diferentes características del índice. Resultados La diferencia entre la frecuencia cardíaca del modelo propuesto y la frecuencia cardíaca real es de 0,11, la diferencia entre la frecuencia cardíaca del modelo y la frecuencia cardíaca real es de 4,28, y la diferencia entre la frecuencia cardíaca del modelo y la frecuencia cardíaca real es de 2,84, y la precisión del modelo propuesto es la más alta. Conclusión El modelo propuesto en este trabajo puede analizar con precisión los efectos de diferentes frecuencias de ejercicio aeróbico en la resistencia cardiorrespiratoria. Nivel de evidencia II; Estudios terapéuticos - Investigación de resultados.

Context Sensing System Analysis for Privacy Preservation Based on Game Theory.

In a context sensing system in which a sensor-equipped mobile phone runs an unreliable context-aware application, the application can infer the user's contexts, based on which it provides personalized services. However, the application may sell the user's contexts to some malicious adversaries to earn extra profits, which will hinder its widespread use. In the real world, the actions of the user, the application and the adversary in the context sensing system affect each other, so that their payoffs are constrained mutually. To figure out under which conditions they behave well (the user releases, the application does not leak and the adversary does not retrieve the context), we take advantage of game theory to analyze the context sensing system. We use the extensive form game and the repeated game, respectively, to analyze two typical scenarios, single interaction and multiple interaction among three players, from which Nash equilibriums and cooperation conditions are obtained. Our results show that the reputation mechanism for the context-sensing system in the former scenario is crucial to privacy preservation, so is the extent to which the participants are concerned about future payoffs in the latter one.

[Identification of novel KIT gene mutations in two Chinese families with piebaldism].

OBJECTIVE: To screen for potential mutations of KIT gene for two Chinese families affected with piebaldism in order to facilitate genetic counseling and assisted reproduction. METHODS: Peripheral blood samples were collected from 2 patients of family 1 and the proband and 3 unaffected members of family 2 for the extraction of DNA and RNA. PCR-sequencing and reverse transcription PCR-sequencing were used to screen KIT mutations. RESULTS: All of the patients from family 1 were found to carry heterozygous IVS12+2-+7delinsACATCTTTA, a splicing mutation undocumented in the human gene mutation data base (HGMD) database. This mutation has resulted in c.1765-1779del in cDNA and p.Gly592Ala/del:E12, which has led to skipping of exon 12 and no expression of cDNA. The proband from family 2 has carried a heterozygous c.2401A>C mutation in KIT gene. The same mutation was not found in unaffected members. CONCLUSION: We have attained definite diagnosis for both families, which has facilitated genetic counseling and assisted reproduction for our patients and their family members.

[A de novo partial 5p deletion and cryptic 18p duplication detected by SNP-Array in a boy featuring Cri du Chat syndrome].

OBJECTIVE: To determine the karyotype of a boy suspected to have Cri du Chat syndrome with severe clinical manifestations, and to assess the recurrence risk for his family. METHODS: High-resolution GTG banding was performed to analyze the patient and his parents. Fluorescence in situ hybridization (FISH) with Cri du Chat syndrome region probe as well as subregional probes mapped to 5pter, 5qter, 18pter, 18qter, and whole chromosome painting probe 18 was performed to analyze the patient and his parents. In addition, single nucleotide polymorphism-based arrays (SNP-Array) analysis with Affymetrix GeneChip Genome-wide Human SNP Nsp/Sty 6.0 were also performed to analyze the patient. RESULTS: Karyotype analysis indicated that the patient has carried a terminal deletion in 5p. FISH with Cri du Chat syndrome region probe confirmed that D5S23 and D5S721 loci are deleted. SNP-Array has detected a 15 Mb deletion at 5p and a 2 Mb duplication at 18p. FISH with 5p subtelomeric probes and 18p subtelomeric probe further confirmed that the derivative chromosome 5 has derived from a translocation between 5p and 18p, which has given rise to a 46,XY,der(5)t(5;18)(p15.1;p11.31)dn karyotype. CONCLUSION: A de novo 5p partial deletion in conjunction with a cryptic 18p duplication has been detected in a boy featuring Cri-du-Chat syndrome. His parents, both with negative findings, have a low recurrence risk. For its ability to detect chromosomal imbalance, SNP-Array has a great value for counseling of similar patients and assessment of recurrence risks.

[Common mutations of congenital adrenal hyperplasia are also the hotspots for new mutations].

OBJECTIVE: To discuss the genetic diagnosis of congenital adrenal hyperplasia (CAH) and investigate the resource of gene mutations in CAH. METHOD: Enzymatic methods with restriction endonucleases that specifically recognized the mutation sites were used to detect the gene mutations in patients with CAH and their relatives. Polymerase chain reaction and direct sequencing were used to identify the mutations in 21-hydroxylase gene, and short tandem repeat (STR) typing was used to determine the sources of the mutations. RESULTS: One CAH patient had two known mutations in 21-hydroxylase gene, namely the I2g and I172N mutations. The former mutation was inherited from the biological mother and the latter was not inherited. CONCLUSION: The 9 common mutations of CAH are also the hotspots for new mutations.

[Mutation screening and prenatal diagnosis of tuberous sclerosis complex].

OBJECTIVE: To screen mutations of tuberous sclerosis complex (TSC) patients to confirm a clinical diagnosis of TSC, and to perform prenatal diagnosis for families with mutations. METHODS: In this study, PCR-denaturing high-performance liquid chromatography(DHPLC), supplemented with sequencing when necessary, was used to screen TSC1 and TSC2 mutations in 21 patients from 19 pedigrees visited author's hospital in the last five years. For novel mutations, one hundred unrelated healthy individuals were screened to exclude the possibility of polymorphism. RESULTS: Seventeen different mutations were found in 21 patients of 19 pedigrees with 13 being novel mutations, including c. 2672delA, c. 2672insA of TSC1 gene and c.4918insCGCC, c.1143delG, Intron27+1 G>A, c.1957-1958delAG, Intron5+1 G>A, c.910insCT, c.2753 C>G, c.4078dupAGCAAGTCCAGCTCCTC, Intron 11 -1 G>A, Intron 14+1 G>A, c.684 C>A of TSC2 gene, indicating a high frequency of de novo mutations in TSC. Three of these mutations were in the TSC1 gene (N762S, c.2672insA and c. 2672delA), while all remaining 14 were in the TSC2 gene. Prenatal diagnosis for TSC was performed for 7 fetuses from these pedigrees. The six fetuses that tested negative for TSC mutations were carried to term and, to date, none of these children has shown symptoms of TSC. CONCLUSION: Author's data showed that a mutation detection rate of tuberous sclerosis was 89.5%(17/19) among patients in author's hospital. The ratio of TSC2 and TSC1 mutations was about 1:1 in the familial cases, but TSC2 mutation was more common than TSC1 mutation in sporadic cases. Author's data demonstrated that birth of TSC children for those with familial history of TSC could be prevented through prenatal diagnosis.

[Mutation screening and prenatal diagnosis of a pedigree with Glanzmann's thrombasthenia].

OBJECTIVE: Mutation screening was performed in a pedigree of Glanzmann's thrombasthenia (GT) and prenatal diagnosis was performed. METHODS: In this study, reverse transcription-PCR-sequencing and PCR-sequencing, as well as restriction fragment length polymorphism(RFLP) and A/T-cloned-sequencing, were used to screen the ITGA2B and ITGB3 mutation in a pedigree with Glanzmann's thrombasthenia in the RNA and DNA level. Prenatal diagnosis was performed for this pedigree. RESULTS: Deletion of 99 bps was found in the cDNA of the patient in the pedigree, leading to deletion of 33 codons (from codon 160 to 192). After genomic analysis, the patient was found to be a compound heterozygote of c.374C to G mutation and intron 4(IVS-4) + 5 G to C mutation. The two mutations were inherited from the parents. IVS-4 + 5 G to C mutation was a point mutation in the splice site, while c.374C to G mutation was out of the splice site. But both of them resulted in the same splice pattern in RNA. The two mutations were novel mutations which have not been reported in Human Gene Mutation Database (HGMD) and the mutation data base of Glanzmann's thrombasthenia. The results of ITGB3 gene screening is normal in the proband and his parents. CONCLUSION: Two novel mutation, c.374C to G and IVS-4 + 5 G to C were found in this study, which might be the cause of GT in the pedigree.

[Mutation screening of the F VIII gene in 10 hemophilia A families].

OBJECTIVE: To identify the F VIII gene mutations of patients and suspected female carriers in 10 Hemophilia A (HA) families, and to guide the prenatal diagnosis. METHODS: PCR, denaturinghigh performance liquid chromatogramphy (DHPLC) and DNA sequencing technologies were applied to screen the F VIII gene of 8 HA patients and 12 suspected female carriers in the 10 families. Linkage analysis was performed by using St 14(DXS 52), intron 13 (CA)n and EX18/Bcl I of the F VIII gene in the HA families. In prenatal diagnosis, we screened the same mutation found in the patients. PCR-restriction fragment length polymorphism was applied to detect the new missense mutations of F VIII gene in 100 unrelated healthy individuals to exclude the possibility of polymorphism. RESULTS: Five missense mutations, 3 frameshift mutations, 2 nonsense mutations and 2 single nucleotide polymorphism (SNP) were identified in 10 the HA families. Among them, c.878A to G, c.1015A to G, c.6870G to T, c.1282delA, c.3072_3073insT, c.4880_4881insA and c.5000G to A were novel mutations or polymorphism. No missense mutations c.878A G, c.1015A to G and c.6870G to T, were found in the 100 healthy unrelated controls. (2) Nine suspected female carriers were confirmed at the gene level. (3) X risk chromosome could be determined to in 4 HA families by genetic linkage analysis. (4) Among the four fetuses for prenatal diagnosis, 2 were normal, 1 was carrier and the remaining 1 was a patient. CONCLUSION: Six novel mutations, i.e., c.878A to G, c.1015A to G, c.6870G to T, c.1282delA, c.3072_3073insT and c.4880_4881insA, were identified in this study. PCR, DHPLC and DNA sequencing could be used to screen the gene mutations of HA patients, to carry out carrier detection and prenatal diagnosis of HA families efficiently, by combining with restriction endonuclease analysis and genetic linkage analysis.

[Identification of a cryptic 1p36.3 microdeletion in a patient with Prader-Willi-like syndrome features].

OBJECTIVE: To determine the karyotype of a patient with Prader-Willi-like syndrome features. METHODS: Chromosomal high resolution banding was carried out to analyze the karyotype of the patient, and methylation-specific PCR was used to analyze the imprinting region of chromosome 15. Subtelomeric region was screened by multiplex ligation-dependent probe amplification (MLPA), and fluorescent in situ hybridization (FISH) and real-time quantitative PCR were further performed to identify the deleted region. RESULTS: No abnormality was discovered by high resolution karyotype analysis and methylation-specific PCR studies. MLPA analysis showed that the patient had a deletion of 1p subtelomeric area, which was confirmed by FISH analysis. The deleted region was shown within a 4.2 Mb in the distal 1p by 3 BAC FISH probes of 1p36 combined with real-time PCR technique. Family pedigree investigation showed the chromosome abnormality was de novo. Therefore, partial monosomy 1p36 was likely responsible for the mental retardation of the patient. CONCLUSION: Molecular cytogenetic techniques should be performed to those patients with Prader-Willi-like syndrome features, to determine their karyotypes.

Mutation Analysis and Prenatal Exclusion of Fibrodysplasia Ossificans Progressiva in a Chinese Fetus.

Aims: Fibrodysplasia ossificans progressiva (FOP) is a rare and severely disabling autosomal dominant disorder characterized by congenital malformations of the great toes and progressive postnatal heterotopic ossification. A point mutation in the activin receptor IA (ACVR1) gene is the cause of FOP. Most of the reported cases of FOP are sporadic and caused by de novo mutations; however, some rare cases can also result from parental germline mosaicism associated with a greater risk of recurrence in successive pregnancies. Therefore, once the pathogenic mutation has been identified in the proband, it is relative cheaper and important to perform prenatal diagnostic tests to exclude the recurrence risk of FOP in subsequent pregnancies. In this study, we first investigated the mutation in the ACVR1 gene in a Chinese FOP patient and then performed prenatal tests to exclude the risk of recurrence in the patient's unborn sibling. Methods: A couple visited our clinic with their 4-year-old son, who was clinically diagnosed with FOP, for genetic counseling. Genetic testing was performed by amplifying all the nine exons of the ACVR1 gene using the conventional polymerase chain reaction. Further, DNA sequencing was used to determine the mutation based on the results of a mutation screening using denaturing high-performance liquid chromatography. Subsequently, a prenatal test was performed using the same technique as that used for the proband. Results: A recurrent single nucleotide mutation c.617 G>A (R206H) of the ACVR1 gene was identified in the patient; however, both the parents had a normal ACVR1 gene. Prenatal tests showed that the fetus did not carry the pathogenic mutation. Conclusion: The results confirmed that a recurrent single nucleotide mutation c.617 G>A (R206H) was the genetic cause of FOP and explored the utility of prenatal testing in excluding the risk of recurrence in the successive pregnancy.

Crypt Y chromosome fragment resulting from an X;Y translocation in a patient with premature ovarian failure.

OBJECTIVE: To identify a cryptic Y chromosome fragment that resulted from a X;Y translocation in a patient with premature ovarian failure (POF) and analyze the karyotype-phenotype correlation. DESIGN: Case report. SETTING: A university-based reproductive medicine center. PATIENT(S): A 33-year-old woman with POF. INTERVENTION(S): Karyotyping analysis, comparative genomic hybridization, fluorescence in situ hybridization, and polymerase chain reaction (PCR) analysis for the patient. MAIN OUTCOME MEASURE(S): Karyotype determination of the patient. RESULT(S): The patient was suspected to carry an abnormal X chromosome by traditional cytogenetic analysis. A Y chromosome hybridization signal was found in the patient's genome by comparative genomic hybridization analysis. The fluorescence in situ hybridization result showed that the Y chromosome material resulted from a translocation between Xq and Yq. Using the specific sequence-tagged sites, the breakpoints on the X and Y chromosomes were located at Xq26.3 and Yq11.223, respectively. Combined with chromosome G banding and C banding, the karyotype of the patient was determined as 46,X,der(X)t(X;Y) (q26.3;q11.223). CONCLUSION(S): The advanced molecular cytogenetic techniques are helpful to detect cryptic chromosome aberrancies in patients with POF. This rare case supports that Xq26-q28 is the critical region of POF, and is helpful to analyze the risk of gonadoblastoma in patients with POF with Y chromosomal material.

Molecular cloning and characterization of a novel transcript variant of Mtsarg1 gene.

Mtsarg1 (Mus musculus testis and spermatogenesis cell apoptosis-related gene 1) gene with 1103 bp in full length had been cloned previously (GenBank accession number: AF399971, 2002; re-designated as Spata3, Mus musculus spermatogenesis-associated 3, 2007). In the present study, we identified a novel transcript variant of Mtsarg1, named Mtsarg1-beta which is 887 bp in length (GenBank accession numbers: EU259321 and EF546784, 2007, designated as Spata3 variant 4) by reverse transcription-polymerase chain reaction (RT-PCR), cloning and sequencing. Mtsarg1-beta which has high similarity with Mtsarg1 contains an entire open reading frame of 417 bp encoding a protein consisting of 138 amino acids. Mtsarg1-beta protein is a non-secretory protein with a theoretic molecular mass around 14.79 kD and an isoelectric point of 9.74, which shares the 100 N-terminal amino acids with Mtsarg1 followed by 38 amino acids differing from Mtsarg1. Multi-tissue RT-PCR results and Northern blot analysis for adult DBA/2 mice showed that Mtsarg1-beta and Mtsarg1 mRNAs were specifically expressed in testis at high level. RT-PCR results also showed that Mtsarg1-beta and Mtsarg1 mRNAs were not expressed in mouse GC-1 spermatogonia. In situ hybridization revealed that both Mtsarg1 and probably Mtsarg1-beta mRNAs were mainly expressed in mouse spermatocytes. Subcellular localization analysis suggested that Mtsarg1 protein was mainly localized in nucleus while Mtsarg1-beta protein was mainly localized in cytoplasm. All these results indicate that Mtsarg1 and Mtsarg1-beta may play an important role in mouse testicular function and in spermatocyte development.

[Mutation screening and prenatal diagnosis of Wilson's disease by denature high performance liquid chromatography].

OBJECTIVE: To study the clinical application of denature high performance liquid chromatography (DHPLC) technique on mutation screening and prenatal diagnosis for Wilson's disease (WD). METHODS: Genomic DNA of the probands with Wilson's disease and their parents from 6 families was subjected to polymerase chain reaction (PCR) for the 21 exons and the 5' untranslated region of ATP7B gene. Mutation screening of the PCR products was performed by DHPLC. The abnormal peaks were confirmed by further sequencing analysis. Based on the successful gene diagnosis for the patients, prenatal diagnosis was performed in 4 families, including 1 twin and 3 singletons. RESULTS: Five disease-causing mutations and 8 polymorphisms were found in the 6 probands by DHPLC and sequencing. The parents were carriers with the same mutation as their affected children. Prenatal diagnosis showed that two pregnancies were abnormal, including a twin pregnancy with compound heterozygote for Arg778Leu and IVS4-1G>C mutation, and a single pregnancy with a compound heterozygote for Ser975Tyr and Pro992Leu mutations. These two pregnancies were terminated after genetic counseling. Another two pregnancies included a singleton carrier with Ser975Tyr mutation and a normal genotype fetus, respectively. These two pregnancies were continued and the babies were healthy. CONCLUSION: DHPLC is a powerful tool in prenatal diagnosis as well as in postnatal diagnosis.

[Mutation screening of the TYR and P gene in three patients with oculocutaneous albinism].

OBJECTIVE: To identify the mutations of the tyrosinase gene (TYR) and P gene in patients with oculocutaneous albinism (OCA). METHODS: Polymerase chain reaction (PCR) and denaturing high performance liquid chromatography (DHPLC) were applied to detect the mutations in all exons of TYR gene and P gene. Then DNA sequencing and restriction endonuclease analysis were used to confirm the mutations detected by DHPLC. Novel mutations were screened in 100 unrelated persons with normal phenotypes to exclude the possibility of polymorphism. RESULTS: Two mutations were detected in the P gene of the three patients and none in TYR gene. Heterozygous mutation of T450M in exon 13 of the P gene was detected in patient 1. Patient 2 had a heterozygous mutation of T450M in exon 13 and a heterozygous mutation of G775R in exon 23 of the P gene. Patient 3 had a heterozygous mutation of G775R as well. Restriction endonuclease analysis of the P gene exon 13 showed that the Oli I site had partly disappeared resulting from the heterozygous mutation T450M in patient 1 and patient 2, but not in 100 unrelated individuals. The heterozygous mutation T450M is a novel mutation. CONCLUSION: Gene diagnosis of OCA can be carried out effectively by combining PCR, DHPLC, DNA sequencing and restriction endonuclease analysis.

[Mutation detection of PKD1 gene in patients with autosomal dominant polycystic kidney diseases].

OBJECTIVE: To detect gene mutation in the patients with autosomal dominant polycystic kidney disease (PKD). METHODS: Polymerase chain reaction (PCR)-denaturing high-performance liquid chromatography (DHPLC) analyses were performed in 3o single copy region of PKD 1 gene (PKD1). DNA sequencing were carried out on PCR products with abnormal peak shape afterwards. RESULTS: A new nonsense mutation (C11901A in exon 42 of PKD1 was identified to cause serine in position 3897 turning to a stop codon. A missense mutation, C10737T, was detected in exon 35 which caused threonine in position 3509 turn to methionine. Two kinds of samesense mutation, G11824A and C11860T in exon 42, were found in normal control. CONCLUSION: PKD1 mutation were detected successfully by PCR-DHPLC. A new nonsense mutation, a missense mutation and two polymorphisms are identified in this study.

[Delineating a supernumerary marker chromosome by combining several cytogenetic and molecular cytogenetic techniques].

OBJECTIVE: To characterize a supernumerary marker chromosome (SMC) by comparative genomic hybridization (CGH), fluorescence in situ hybridization (FISH) and traditional cytogenetic techniques, and to explore the clinical application of these techniques in delineating de novo marker chromosomes. METHODS: A mental retardation patient received chromosome test by ordinary G banding. CGH and FISH techniques were used to analyze the origin of the de novo SMC, and N banding technique and C banding techniques were used to analyze the SMC structure. The phenotypic effects of the SMC were analyzed after the karyotype was determined. RESULTS: By G banding technique, the patient was showed to have a mosaic karyotype with SMC: mos.47, XX, +mar [31]/48, XX, +2mar[29]. CGH analysis showed a gain of 15q11 --> q14, and the result was confirmed by FISH with chromosome 15 painting probe. The further FISH analysis showed the SMC had two signals with UBE3A probe for detecting Prader-willi syndrome/Angelman syndrome (PWS/AS). N banding and C banding analysis showed the SMC had a double satellite and double centromere, respectively. Combined with the above results, the karyotype of the patient was: mos.47, XX, +der (15) (pter --> q14::q14 --> pter) [31]/48, XX, +2der (15) (pter --> q14::q14 --> pter) [29]. ish der(15)(WCP15+, UBE3A++, PML-). CONCLUSION: CGH is a valuable method to detect imbalanced chromosomal rearrangement. Combined with FISH and the traditional cytogenetic technique, it provides a valuable technique platform for characterizing the structure of the de novo SMC, and a basis for exploring the relation between karyotype and phenotype, prognosis and recurrent risk.