Molecular genetic analysis of primary renal epithelial tumours with granular oncocytic cytoplasms / Análisis genético molecular de tumores epiteliales renales primarios con citoplasma oncocítico granular

SUMMARY: The group of primary renal tumours with granular-oncocytic cytoplasm is a very heterogeneous group, in its histological origin and biological behavior resulting in many diagnostic problems. In this study 57 renal epithelial tumours with granular oncocytic cells were analyzed using fluorescence in situ hybridisation (FISH), array comparative genomic hybridisation (aCGH) and polymerase chain reaction (PCR). The results of analysis in renal oncocytoma (RO) did not indicate the presence of the gene mutations or chromosomal abnormalities. Sporadic renal hybrid oncocytic/chromophobe tumours (HOCT) had multiple numerical aberrations of chromosomes 1, 2, 6, 9, 10, 13, 17, 20, 21 and 22. This type of tumour had no mutations in the VHL, c-kit, PDGFRA, and FLCN genes. Oncocytic papillary renal cell carcinoma (O-PRCC) had numerical abnormalities of chromosomes 7 and 17 and the loss of the Y chromosome. Cytogenetic analysis of 20 pigmented microcystic chromophobe renal cell carcinomas (PMChRCC) showed monosomy as the most frequent aberration in all analyzed chromosomes 1, 2, 5, 10, 13, 17 and 21. One case of chromophobe renal cell carcinoma (ChRCC) with hyaline globules had a mutation in the distal part of exon 3 of the VHL gene. Absence of genetic disorders in usual RO is common result, but we have established absence of genetic disorders even in rare variants. Variety of genetic alterations detected in sporadic renal HOCT proves it to be a separate entity, not a variant of ChRCC, while PMChRCC is an uncommon variant of ChRCC. O-PRCC is a subtype of papillary renal cell carcinoma.

RESUMEN: El grupo de tumores renales primarios con citoplasma granular-oncocítico es un grupo muy heterogéneo, en su origen histológico y comportamiento biológico, resultando en problemas de diagnóstico. En el estudio se analizaron 57 tumores epiteliales renales con citoplasma oncocítico granular mediante hibridación fluorescente in situ (FISH), hibridación genómica comparativa de matriz (aCGH) y reacción en cadena de la polimerasa (PCR). Los resultados del análisis en oncocitoma renal (RO) no indicaron la presencia de mutaciones genéticas ni anomalías cromosómicas. Los tumores oncocíticos / cromófobos híbridos renales esporádicos (HOCT) tenían múltiples aberraciones numéricas de los cromosomas 1, 2, 6, 9, 10, 13, 17, 20, 21 y 22. No se observaron mutaciones en este tipo de tumor en el VHL, c-kit, PDGFRA y genes FLCN. El carcinoma de células renales papilar oncocítico (O-PRCC) tenía anomalías numéricas de los cromosomas 7 y 17 y la pérdida del cromosoma Y. El análisis citogenético de 20 carcinomas de células renales cromófobos microquísticos pigmentados (PMChRCC) mostró que la monosomía era la aberración más frecuente en todos los cromosomas analizados 1, 2, 5, 10, 13, 17 y 21. Un caso de carcinoma de células renales cromófobo (CCRc) hialino tenía una mutación en la parte distal del exón 3 del gen VHL. La ausencia de trastornos genéticos en la OI habitual es un resultado común, pero hemos establecido la ausencia de trastornos genéticos incluso en variantes raras. Varias alteraciones genéticas detectadas en esporádica HOCT renal demuestran que es una entidad separada, no una variante de ChRCC, mientras que PMChRCC es una variante poco común de ChRCC. O-PRCC es un subtipo de carcinoma papilar de células renales.

Chinese Siblings with Prader-Willi Syndrome Inherited from Their Paternal Grandmother

Background: Prader-Willi syndrome (PWS) is a complex neurobehavioral disorder causedby failure of expression of paternally inherited genes in the PWS region of chromosome 15.Case characteristics: Two siblings who both met the inclusion criteria for clinical diagnosisof PWS during neonatal period. Outcome: Molecular genetic analysis demonstrated a 417-kb microdeletion within the 15q11.2 region inherited from siblings’ paternal grandmother,involving key genes of PWS, except for UBE3A, which may explain why their father andpaternal grandmother had a normal phenotype. Conclusion: The findings may be helpfulfor better understanding of the underlying mechanism of this rare imprinting defect

Next generation sequencing based molecular genetic analysis of a Chinese Han family with autosomal retinitis pigmentosa / 中华实验眼科杂志

Background Retinitis pigmeutosa (RP) is a progressive inheritance disease.It is characterized by highly genetical and phenotypical heterogeneity.With the rapid development of genomics,new methods are applied to the genetic screening of RP.Objective This study was to characterize the clinical features of a Chinese family with autosomal RP and to screen the candidate genes.Methods Twelve members from this family were included in the study.All participants underwent complete ophthalmologic examinations.Targeted-capture next generation sequencing (NGS) based molecular genetic analysis was performed on two patients of this RP family(Ⅱ5,Ⅱ 7).The DNA sample from the two patients was separately sequenced using custom capture gene chip,which includes 59 retinal disease genes.The sequencing results were analyzed by bioinformatics technology.Identified variations were verified in the rest family members by PCR and Sanger sequencing.This study was approved by Ethic Committee of West China Hospital,and informed consent was obtained from the subjects.Results Four members of this family were diagnosed as RP,and the rest were asymptomatic.Missense mutation (c.3065T＞C,p.Phe1022Ser) in USH2A and missense mutation (c.1699G＞A,p.Ala1319Gly) in PDE6A were found in two patients (Ⅱ 5 and Ⅱ7).The variants were not co-segregated with the phenotype of this family.The causative mutation was not found by the targeted-capture NGS based eye disease chip,but it ruled out a large number of candidate genes for RP.Conclusions Our study suggests that targeted-capture NGS based eye disease chip can quickly detect mutations in known RP genes.It can be a new applicable and efficient method for molecular genetic analysis of ocular disease.

Research progress of molecular genetic analysis in Schistosoma variation / 中国血吸虫病防治杂志

The development of molecular biology techniques makes important contributions to the researches of heritable varia-tion of Schistosoma. In recent years,the molecular genetic analysis in the Schistosoma variation researches mainly includes the re-striction fragment length polymorphism(RFLP),random amplified polymorphism technology(RAPD),microsatellite anchored PCR(SSR-PCR),and polymerase reaction single-strand conformation polymorphism(PCR-SSCP). This article reviews the re-search progress of molecular genetic analysis in Schistosoma variation in recent years.

Molecular Genetic Analysis of Partial 9p Trisomy in Two Chinese Families with Mental Retardation and Facial Anomaly / 华中科技大学学报（医学）（英德文版）

Mental retardation is defined by significant limitations in intellectual function and adaptive behavior that occur before 18 years of age.Many chromosomal diseases come with mental retardation.We reported two Chinese families with partial trisomy 9p and other chromosome partial monosomy,clinical features of mental retardation and mild facial and pinkie anomalies.In the family 1,we showed that the proband carried a trisomy 9p21.3→pter and monosomy 21q22.3→qter by using fluorescence in situ hybridization analysis.Molecular genetic analysis defined the precise breakpoint on chromosome 9p between markers D9S1846 and D9S171,an interval of about 2.9 Mb on 9p21.3,and the breakpoint on chromosome 21q between markers D21S1897 and D21S1446,a region of about 1.5 Mb on 21q22.3.In the family 2,a patient with trisomy 9p21.3→pter and monosomy 5p15.33→pter,and a de novo maternal balanced translocation between chromosomes 5 and 9 was identified in his mother.Cytogenetic and molecular genetic analysis defined the precise breakpoints on chromosome 9p21.3 and chromosome 5p15.33.Further clinical investigation found that any individual had no refractoriness eczema disease except the proband in this family.These results further implicate that trisomy 9p is associated with mental retardation,and that there may be key gene duplication on chromosome 9p21.3→9pter responsible for mental retardation and mild facial anomaly.This result has been applied successfully in prenatal diagnosis of the second family.

Molecular Genetic Analysis of Cis-AB with Cytosine at nucleotide position 467

BACKGROUNDS: The molecular genetic characteristcs of cis-AB blood group have shown that its allele had C, G, C and C at nucleotide positions (nps) 526, 703, 796 and 803, respectively. And all cis-AB analysed and reported molecular genetically in Korea and Japan had T at np 467 (leucine at amino acid position 156). We report a first case of cis-AB with C at np 467 (proline at amino acid position 156). METHODS: Genomic DNA was extracted from peripheral blood of cis-AB patient and amplified by DS1/DS2 and DS3/DS4 allele-specific primers. After PCR, we analysed nps 261, 467, 526, 646, 703, 796, and 803 by restriction digestion, autoradiography and automatic sequencing. RESULTS: PCR-RFLP with DS1/DS2 primers and restriction enzyme KpnI showed that cis-AB had an 0 allele. The results of genomic sequencing, autoradiography and restriction digestion showed that cis-AB allele at nps 467, 526, 646, 703, 796 and 803 had C, C, T, G, C and C, respectively. CONCLUSION: This cis-AB showed characteristic molecular genetic features at nps 526, 703, 796, and 803. And this is a first case of A(Pro) cis-AB with C at np 467. (Korean J Blood Transfusion 10(1): 13-19, 1999)

Molecular Genetic Analysis of Cis-AB with Cytosine at nucleotide position 467 / 대한수혈학회지

BACKGROUNDS: The molecular genetic characteristcs of cis-AB blood group have shown that its allele had C, G, C and C at nucleotide positions (nps) 526, 703, 796 and 803, respectively. And all cis-AB analysed and reported molecular genetically in Korea and Japan had T at np 467 (leucine at amino acid position 156). We report a first case of cis-AB with C at np 467 (proline at amino acid position 156). METHODS: Genomic DNA was extracted from peripheral blood of cis-AB patient and amplified by DS1/DS2 and DS3/DS4 allele-specific primers. After PCR, we analysed nps 261, 467, 526, 646, 703, 796, and 803 by restriction digestion, autoradiography and automatic sequencing. RESULTS: PCR-RFLP with DS1/DS2 primers and restriction enzyme KpnI showed that cis-AB had an O allele. The results of genomic sequencing, autoradiography and restriction digestion showed that cis-AB allele at nps 467, 526, 646, 703, 796 and 803 had C, C, T, G, C and C, respectively. CONCLUSION: This cis-AB showed characteristic molecular genetic features at nps 526, 703, 796, and 803. And this is a first case of A(Pro) cis-AB with C at np 467.