Comparative analysis of differentially expressed genes for biosynthesis of active ingredients in fruits of different cultivars of Lycium barbarum L. based on transcriptome sequencing / 生物工程学报

To explore the differentially expressed genes (DEGs) related to biosynthesis of active ingredients in wolfberry fruits of different varieties of Lycium barbarum L. and reveal the molecular mechanism of the differences of active ingredients, we utilized Illumina NovaSeq 6000 high-throughput sequencing technology to conduct transcriptome sequencing on the fruits of 'Ningqi No.1' and 'Ningqi No.7' during the green fruit stage, color turning stage and maturity stage. Subsequently, we compared the profiles of related gene expression in the fruits of the two varieties at different development stages. The results showed that a total of 811 818 178 clean reads were obtained, resulting in 121.76 Gb of valid data. There were 2 827, 2 552 and 2 311 DEGs obtained during the green fruit stage, color turning stage and maturity stage of 'Ningqi No. 1' and 'Ningqi No. 7', respectively, among which 2 153, 2 050 and 1 825 genes were annotated in six databases, including gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG) and clusters of orthologous groups of proteins (KOG). In GO database, 1 307, 865 and 624 DEGs of green fruit stage, color turning stage and maturity stage were found to be enriched in biological processes, cell components and molecular functions, respectively. In the KEGG database, the DEGs at three developmental stages were mainly concentrated in metabolic pathways, biosynthesis of secondary metabolites and plant-pathogen interaction. In KOG database, 1 775, 1 751 and 1 541 DEGs were annotated at three developmental stages, respectively. Searching the annotated genes against the PubMed database revealed 18, 26 and 24 DEGs related to the synthesis of active ingredients were mined at the green fruit stage, color turning stage and maturity stage, respectively. These genes are involved in carotenoid, flavonoid, terpenoid, alkaloid, vitamin metabolic pathways, etc. Seven DEGs were verified by RT-qPCR, which showed consistent results with transcriptome sequencing. This study provides preliminary evidences for the differences in the content of active ingredients in different Lycium barbarum L. varieties from the transcriptional level. These evidences may facilitate further exploring the key genes for active ingredients biosynthesis in Lycium barbarum L. and analyzing their expression regulation mechanism.

Prenatal diagnosis of a case with 46,XX,del(4),dup(21) / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the genetic cause and prognosis of a fetus with a rare karyotype.</p><p><b>METHODS</b>Fluorescence in situ hybridization (FISH) was used for verifying a structural chromosomal abnormality detected by conventional karyotyping analysis. Whole genome DNA microarray was used to analyze copy number variations carried by the fetus.</p><p><b>RESULTS</b>The fetus was found to have a 46,XX,dup(21)(?q21q22) karyotype, which was verified by FISH analysis as repetition of chromosome 21 region, namely nuc ish 21q22×3. Whole genome DNA microarray confirmed that there was a 17.87 Mb duplication in the 21q21.3q22.3 region, which involved GATA1, JAK2 and ALL genes and spanned the Down syndrome region. The genes are implicated in craniofacial abnormalities, cardiac abnormalities, mental retardation, growth retardation, limb abnormalities. In addition, there was also an 8.43 Mb deletion in the 4p16.1p16.3 region, which involved FGFR3, LETM1, WHSC1 and WHSC2 and other 64 OMIM genes and spanned the Wolf-Hirschhorn syndrome region. The genes are implicated in growth retardation, craniofacial abnormalities, cardiac abnormalities, mental retardation, and hypotonia. After consultation, the family chose to terminate the pregnancy at 25th week of gestation.</p><p><b>CONCLUSION</b>FISH can help to verify structural chromosome abnormalities suspected by conventional karyotyping analysis. Combined with whole genome microarray, these can determine copy number variation and its region containing the disease genes, and facilitate clinical analysis of the fetus.</p>

Genetic and prenatal diagnosis of a pregnant women with mental retardation / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To conduct genetic testing and prenatal diagnosis for a pregnant women with growth retardation, severe mental retardation, and a history of adverse pregnancies.</p><p><b>METHODS</b>G-banded chromosome analysis, fluorescence in situ hybridization (FISH), and whole genome DNA microarray were used to analyze the patient and her fetus.</p><p><b>RESULTS</b>The women was found to be a chimera containing two cell lines with 47 and 46 chromosomes, respectively. Both have involved deletion of 18q21.2q23. FISH analysis suggested that the cell line containing 47 chromosomes has harbored a chromosome marker derived from chromosome 15. The marker has contained chromosome 15p involving the SNRPN locus and part of 15q, which gave rise to a karyotype of 47,XX,del18q21.3,+ish mar D15Z1+ SNRPN+[82]/46,XX,del18q21.3[18]. Whole genome DNA microarray confirmed that a 3.044 Mb fragment from 15q11.2q12 was duplicated, which involved NIPA1, SNRPN and other 17 OMIM genes. Duplication of this region has been characterized by low mental retardation, autism, developmental delay. Meanwhile, there was a 17.992 Mb deletion at 18q21.33q23, which contained 39 OMIM genes including TNFRSF11A and PHLPP1. This fragment was characterized by mental retardation, developmental delay, short stature, and cleft palate. Whole genome microarray analysis confirmed that there was a 17.9 Mb deletion at 18q21.33q23, which has been implemented with mental retardation, general growth retardation, short stature, and cleft palate. After genetic counseling, the family decided to terminate the pregnancy at 21st week.</p><p><b>CONCLUSION</b>Combined chromosome karyotyping, FISH, and whole genome DNA microarray can determine the origin of marker chromosomes and facilitate delineation of its correlation with the clinical phenotype.</p>

Detection of a fetus with paternally derived 2q37.3 microdeletion and 20p13p12.2 microduplication using whole genome microarray technology / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To perform prenatal diagnosis for a fetus with multiple malformations.</p><p><b>METHODS</b>The fetus was subjected to routine karyotyping and whole genome microarray analysis. The parents were subjected to high-resolution chromosome analysis.</p><p><b>RESULTS</b>Fetal ultrasound at 28+4 weeks has indicated intrauterine growth restriction, left kidney agenesis, right kidney dysplasia, ventricular septal defect, and polyhydramnios. Chromosomal analysis showed that the fetus has a karyotype of 46,XY,der(2),der(20), t(2;20)(q37.3;p12.2), t(5;15) (q12.2;q25) pat. SNP array analysis confirmed that the fetus has a 5.283 Mb deletion at 2q37.3 and a 11.641 Mb duplication at 20p13p12.2. High-resolution chromosome analysis suggested that the father has a karyotype of 46,XY,t(2;20)(q37.3;p12.2),t(5;15)(q12.2;q25), while the mother has a normal karyotype.</p><p><b>CONCLUSION</b>The abnormal phenotype of the fetus may be attributed to a 2q37.3 microdeletion and a 20p13p12.2 microduplication. The father has carried a complex translocation involving four chromosomes. To increase the chance for successful pregnancy, genetic diagnosis and/or assisted reproductive technology are warranted.</p>

Progress in the studies on the induction of pluripotent stem cells / 国际生物医学工程杂志

The main objective of human tissue engineering is to grow cells in vivo to promote proliferation and differentiation and replace, repair, maintain, or enhance the functions of impaired tissues and organs. Embryonic stem cells are the first selected seed cells in tissue engineering due to their unlimited proliferation capability and the potential to differentiate into different kinds of cells. In this review will give a sketch of recent advances in the preparation of induced pluripotent stem cells, which can differentiate into many other kinds of cells, by applying different transcript factors including Oct3/4, Sox2, c-myc, Klf4, Nanog and LIN28 to somatic ceils via retroviral transduction.

Rapid detected of trisomy 21 syndrome by gene diagnosis techniques / 中华检验医学杂志

Objective Setting a rapid, simple and accurate gene diagnosis method for trisomy 21 syndrome. Methods 4 short tandem repearts (STRs) loci (D21S11, D21S1411, D21S1412, D21S1414) in and near the core region(21q22.1-21q22.2) on 21 chromosome of blooding samples from 8 normal people, 11 cases of trisomy 21 syndrome and 1 case of fetus umbilical core were analyzed and detected by polymerase chain reaction(PCR), polyacrylamide gel electrophories, and silver staining. Results 6/8? 7/8? 3/8?7/8 of normal people showed two bands with a DNA content ratio of 1∶1. 2/8?1/8?5/8?1/8 of normal people showed one band in 4 loci. 7/11?8/11?6/11?7/11 of all patients showed two bands with a DNA content ratio of 2∶1;3/11?2/11?2/11?3/11 showed three bands with a DNA content ratio of 1∶1∶1. 1/11?1/11?3/11?1/11 showed one band. The blooding sample of fetus showed two bands with a DNA content ratio of 2∶1 at 3 loci and one band at 1 locus. 11 patents and 1 fetus were identified as trisomy 21 syndrome by analysis combining with 4 STR loci. Conclusion The 4 selected STR loci have high polymorphism.4 polymorphotic STR is a valuable gene marker for diagnosis of trisomy 21. Trisomy 21 can be diagnosed rapidly and accurately within 24 hours by PCR.