Objective This research is to assess the heterozygosity of Tbx1 gene on 22q11 2 in the patients with conotruncal heart malformation.Methods By fluorescence in situ hybridization (FISH) with partial segment of Tbx1 gene, we examined 22 patients with conotruncal heart malformation including 5 syndromic cases and 17 isolated cases. Northern blot was performed with RNA of 50 human tissues.Results Two of 5 syndromic patients had chromosome 22q11 2 hemizygote microdeletion of the Tbx1 gene, while 17 isolated patients did not show such deletion. Northern blot showed that there were Tbx1 gene positive expression in skeletal muscle, testis, lung and fetal heart.Conclusions Our study suggests that Tbx1 gene may be one of the pathogenic gene related to CATCH22.Association of the Tbx1 gene and conotruncal heart teratogenic gene is to be further detected in gene mutation of patients without heterozygosity deletion.

Many somatic cell typos were obtained by in vitro differentiation or teratoma formation of human embryonic stem ceLls (hESCs). However, it is unclear whether specific cell types can be obtained from hESCs-derived teratoma. It was reported that many kinds of cells, including neural progetfitor/precursor cells (NPCs) and mesenchymal stem cells (MSCs) were isolated efficiently from the teratoma of hESCs through in vitro selection. The teratoma-derived NPCs and MSCs showed specific characteristics of molecular markers similar to the primary NPCs and MSCs. Moreover, these teratoma-induced NPCs and MSCs can be further induced to differentiate into neurons, astrocytes, or adipose and bone cells, reflecting their inherent multi-potencies. Given that teratoma normally contains a mixture of ectoderm, mesodenn, and endoderm lineage cells at different differentiation stage, it was suggested that hESCs-derived teratoma could be an alternative source to generate a variety of uncommitted progenitor cells or terminally differentiated somatic cells, which may be otherwise difficult to obtain through direct in vitro differentiation.

The gamma-secretase complex mediates the final cleavage of APP to generate the principal component of amyloid plaques in the brains of Alzheimer's disease patients.Four integral membrane proteins (PS,NCT,PEN-2 and APH-1) are essential and sufficient for gamma-secretase activity.To identify the promoter of human nicastrin gene (NCT),its 5' -flanking region has been characterized and a 270 bp fragment containing the TSS (transcription start site) for the promoter activity has been identified.EMSA assays confirmed that all four AP-1 binding sites and two NFAT sites in the NCT promoter region were able to bind relative transcription factors in vitro.Mutations,as well as treatment with PDTC,which adjust the regulatory effect of AP-1 and NFAT,altered NCT promoter activity in both HeLa cells and rat cortical neurons.The results demonstrated that AP-1 and NFAT are involved in the regulation of hNCT transcription and suggest that balanced activation of AP-1 and NFAT ensures a strict temporal and tissue-specific control of NCT transcription.

Harmful substances are produced during the metabolism of alcohol in the human body, which causes liver disease.The mechanism of oxidative stress is the main pathogenesis of alcoholic liver disease, which plays a major role in the occurrence and development of alcoholic liver disease.Alcohol-induced oxidative stress mediated by reactive oxygen species(ROS)/reactive nitrogen species(RNS) brings about liver disease via apoptotic signaling pathway and MAPK signaling pathway, and alleviates liver disease through Nrf2 signaling pathway.In this paper, the studies on the mechanism of oxidative stress in alcoholic liver disease are reviewed.

Objective To isolate and identify the potential binding partners of LRRK2,a gene linked to both dominant familial form and sporadic form of Parkinson's disease,thus to further our knowledge of its function.Methods We used a sequence containing full-length of COR domain and part of ROC and MAPKKK domain as bait.The bait amplified by polymerase chain reaction(PCR) was then cloned into a yeast expression plasmid pGBKT7.After being sequenced and analyzed,pGBKT7-bait was transformed into the yeast strain AH109.Western blot was performed to confirm the expression of pGBKT7-bait in AH109 yeast strain.Then human fetal brain cDNA library was trarnsformed into that yeast strain.which could express pGBKT7-bait fusion protein.The yeast strain which contained pGBKT7-bait and human fetal brain cDNA library was plated on quadruple dropout medium (SD/-Trp/-Leu/-His/-Ade)containing X-a-gal.We retested these positive colonies using 2 independent yeast strains AH109 contained pGBKT7-bait or pGBKT7,respectively.At last,these plasmids isolated from these true positive colonies were analyzed by bioinformatics.Results We obtained 9 true positive colonies,these colonies were sequenced, and we performed sequence Blast in GenBank.Three colonies of the 9 positive colonies were not in open reading-frames.Among other 6 colonies,there were known proteins including spermatid perinuclear RNA-binding protein(STRBP)and BCL2-associated athanogene 5 isoform b(BAG5),as well as unknown proteins including tyrosine phosphatase non-receptor type(PTPN23),1(3)mbt-like 3 isoform b(L3 MBTL3),RALY RNA binding protein-like isoform 1(RALYL),and Homo sapiens mRNA for KIAA1783 protein,partial cds(KIAA 1783).Conclusion True positive colonies of LRRK2 are successfully obtained by the yeast 2-hybrid.Our screened proteins may provide a new research clue for revealing biological functions of LRRK2,pathogenesis of Parkinson's disease,and other neurodegerations.

G mutation of SLC26A4, the parents and the second child were carriers of the same mutation, while the fetus had a wild-type form. Conclusion It is feasible to identify deafness related genes by screening for GJB2 and SLC26A4 mutation, thus providing correct prenatal diagnosis and avoiding deaf delivery of baby.

Objective　To identify the mutation characteristic of STK11 gene in Chinese with Peutz-Jeghers syndrome(PJS) and establish the base of the gene diagnosis of PJS.Methods　STK11 germline mutation was analysed by DNA sequencing in 18 unrelation patients with PJS.Results　Six novel mutations of STK11 gene were detected in six unrelation patients. These mutations will lead to production of truncated protein.Conclusion　STK11 gene mutation accounts for one third of the Chinese with PJS. The content of mutation includes single base substitution or deletion and one or two bases insertion. The mutations were widely found in different regions of the whole coding sequence, and 2/3 of those concentrate in exon 1. Mutation frequency is 66.7% in the family suffering PJS in two or more generations, and 16.7% in the disseminated cases.

Calbindin 2 ; metabolism ; Colon ; metabolism ; Hirschsprung Disease ; metabolism ; Humans

Objective To explore the expression of long noncoding RNA ( lncRNA) RN7SK and its screening value in clinical diagno-sis of gastric cancer. Methods The expression of lncRNA RN7SK in gastric cancer cells and paired gastric cancer tissues were meas-ured by qRT-PCR. The screening efficacy was detected by the receiver operating characteristic ( ROC) curve. Results The expression levels of lncRNA RN7SK in gastric cancer lines SGC-7901(3.91±0.53),MGC-803(3.44±0.29),HGC-27(4.04±0.87)and BGC-823 (4.30±1.13) were markedly higher than that in human gastric mucosal epithelial cell line GES-1, and the difference had statistical sig-nificance(1.02±0.27, t=12.33, 7.48, 7.20 and 4.90, P<0.05). The expression of RN7SK was up-regulated in 85.5％ (47/55) of gastric cancer tissues while down-regulated in the the other 8 tissues ( 14. 5％) . The area under ROC curve ( AUCROC ) of lncRNA RN7SK expression in gastric cancer patients was 0.827 and the 95％ confidence interval (CI) was from 0.746 to 0.907. When the cut-off value was 0.618, the sensitivity and specificity were 0.836 and 0.782 respectively. Conclusion The level of lncRNA RN7SK should be overexpressed in gastric cancer tissues so that it may have high screening efficacy and could be used as a potential molecular marker for the diagnosis of gastric cancer.

In order to study the tumor suppression effect of p53 with CMV enhancer and hTERT promoter mediated by human-derived vector pHrn in liver cancer cell Bel-7402, report plasmid pchEGFP, tumor suppressor plasmids pchp53Arg and pchp53Pro were constructed by inserting expression cassette CMVe+hTERTp+EGFP, CMVe+hTERTp+p53Arg and CMVe+hTERTp+p53Pro into pHrn respectively. 24 h after cell transfection by lipofectamine 2000, GFP expression pattern was analyzed through fluorescence microscope and flow cytometry; RT-PCR and Western blot were taken to study the p53 expression pattern. The cell apoptosis by Hoechst 33258 and Annexin V-FITC/PI staining was also studied. Results show that the expression of GFP and p53 protein in Bel-7402were detected, but apparent cell apoptosis could not be found. The recombinant p53 mediated by human-derived vector could express in Bel-7402, but no significant tumor suppression effect was detected, which might result from the down regulation effect of the wild type p53 on hTERT promoter.