Alternative splicing is a tightly regulated process that contributes to cancer development. CRNDE is a long noncoding RNA with alternative splicing and is implicated in the pathogenesis of several cancers. However, whether deregulated expression of CRNDE is common and which isoforms are mainly involved in cancers remain unclear. In this study, we report that CRNDE is aberrantly expressed in the majority of solid and hematopoietic malignancies. The investigation of CRNDE expression in normal samples revealed that CRNDE was expressed in a tissue- and cell-specific manner. Further comparison of CRNDE expression in 2938 patient samples from 15 solid and hematopoietic tumors showed that CRNDE was significantly overexpressed in 11 malignancies, including 3 reported and 8 unreported, and also implicated that the overexpressed isoforms differed in various cancer types. Furthermore, anti-cancer drugs could efficiently repress CRNDE overexpression in cancer cell lines and primary samples, and even had different impacts on the expression of CRNDE isoforms. Finally, experimental profiles of 12 alternatively spliced isoforms demonstrated that the spliced variant CRNDE-g was the most highly expressed isoform in multiple cancer types. Collectively, our results emphasize the cancer-associated feature of CRNDE and its spliced isoforms, and may provide promising targets for cancer diagnosis and therapy.

OBJECTIVE: To identify potential mutations of F11 gene in a pedigree affected with hereditary coagulation factor XI (FXI) deficiency and explore its molecular pathogenesis. METHODS: Prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), coagulation factor VIII activity (FVIIIC), coagulation factor IX activity (FIXC), coagulation factor XI activity (FXIC), coagulation factor XII activity (FXIIC) and lupus anticoagulation (LA) of the proband and eight family members were determined. FXI antigen (FXIAg) was determined by enzyme-linked immunosorbent assay (ELISA). For the proband, potential mutations in the exons, flanking introns and 5'-, 3'-untranslated regions of the F11 gene were screened by direct DNA sequencing. The results were confirmed by reverse sequencing. Suspected mutations were detected in other family members. ClustalX-2.1-win and four online bioinformatic tools (PolyPhen-2, PROVEAN, SIFT, and Mutation Taster) were used to study the conservation and possible impact of the mutations. The structure of the mutational sites was processed with Swiss-PdbViewer. RESULTS: The propositus had prolonged APTT (69.6 s), whose FXIC and FXIAg were reduced to 6.0% and 10.7%, respectively. Her mother, elder sister, one younger sister, little brother, daughter and son showed slightly prolonged APTT and moderate FXIC and FXIAg levels. Gene sequencing revealed that the propositus carried a heterozygous nonsense mutation c.738G>A (p.Trp228stop) in exon 7 and a heterozygous mutation c.1556G>C (p.Trp501Ser) in exon 13. Her mother, elder sister and daughter were heterozygous for the p.Trp228stop mutation, while one younger sister and little brother and son were heterozygous for p.Trp501Ser. Her husband and the youngest sister were of the wild type. Phylogenetic analysis suggested that Trp501 was highly conserved among all homologous species. The p.Trp501Ser was predicted to be "probably damaging","deleterious", "affect protein function" and "disease causing" corresponding to PolyPhen-2, PROVEAN, SIFT and Mutation Taster. Model analysis demonstrated that the non-polar Trp501 has two benzene rings, forming a hydrogen bond with Gln512 in the wild type. Once substituted by Ser501, the side chain may form another hydrogen bond with the benzene of His396. This may affect the normal space conformation and stability of FXI protein. CONCLUSION The compound heterozygous mutations of the F11 gene probably accounted for the low FXI concentration in this pedigree.
Factor XI ; genetics ; Factor XI Deficiency ; genetics ; Female ; Heterozygote ; Humans ; Male ; Mutation ; Pedigree ; Phylogeny

OBJECTIVE: To detect potential mutations of the coagulation factor Ⅶ (F7) gene in a pedigree affected with hereditary FⅦ deficiency and explore its molecular pathogenesis. METHODS: The FⅦ antigen (FⅦ:Ag) was analyzed by an enzyme-linked immunosorbent assay (ELISA) method. Prothrombin time (PT), FⅦ activity (FⅦ:C) and other coagulant parameters were quantified with an one-stage clotting assay. The F7 gene was amplified by PCR and sequenced. Mutational sites were confirmed by reverse sequencing. Impact of amino acid substitution was assessed using SIFT and PolyPhen-2 software. Structure of the mutant protein was analyzed using Swiss-pdb Viewer software based on the three-dimensional structure in the Protein Data Bank. RESULTS: The propositus had prolonged PT (36.3 s), with FⅦ:C and FⅦ:Ag significantly reduced to 2% and 44%, respectively. Her father, mother, younger sister and daughter had slightly prolonged PT and reduced FⅦ:C (86%-120%). The FⅦ:Ag of her father and younger sister were also reduced. DNA sequencing revealed that the propositus has carried compound heterozygous mutations (Lys341Glu and IVS6-1G>A) of the F7 gene. Her father and younger sister were heterozygous for the IVS6-1G>A mutation, while her mother and daughter were heterozygous for the Lys341Glu mutation. Bioinformatics analysis indicated that Lys341Glu mutation may affect the stability and function of the FⅦ protein. CONCLUSION The Lys341Glu and IVS6-1G>A mutations probably underlie the reduced activity of FⅦ in this pedigree.
Factor VII ; genetics ; Factor VII Deficiency ; genetics ; Female ; Genetic Testing ; Heterozygote ; Humans ; Male ; Mutation ; Pedigree

Objective: To detect potential mutations of the coagulation factor Ⅶ (F7) gene in a pedigree affected with hereditary FⅦ deficiency and explore its molecular pathogenesis. Methods: The FⅦ antigen (FⅦ∶Ag) was analyzed by an enzyme-linked immunosorbent assay (ELISA) method. Prothrombin time (PT), FⅦ activity (FⅦ∶C) and other coagulant parameters were quantified with an one-stage clotting assay. The F7 gene was amplified by PCR and sequenced. Mutational sites were confirmed by reverse sequencing. Impact of amino acid substitution was assessed using SIFT and PolyPhen-2 software. Structure of the mutant protein was analyzed using Swiss-pdb Viewer software based on the three-dimensional structure in the Protein Data Bank. Results: The propositus had prolonged PT (36.3 s), with FⅦ∶C and FⅦ∶Ag significantly reduced to 2% and 44%, respectively. Her father, mother, younger sister and daughter had slightly prolonged PT and reduced FⅦ∶C (86%-120%). The FⅦ∶Ag of her father and younger sister were also reduced. DNA sequencing revealed that the propositus has carried compound heterozygous mutations (Lys341Glu and IVS6-1G>A) of the F7 gene. Her father and younger sister were heterozygous for the IVS6-1G>A mutation, while her mother and daughter were heterozygous for the Lys341Glu mutation. Bioinformatics analysis indicated that Lys341Glu mutation may affect the stability and function of the FⅦ protein. Conclusion The Lys341Glu and IVS6-1G>A mutations probably underlie the reduced activity of FⅦ in this pedigree.

Objective: To analyze the phenotype and F5 gene variant in a pedigree affected with hereditary coagulation factor Ⅴ (FⅤ) deficiency. Methods: All of the exons, flanking sequences, and 5′ and 3′ untranslated regions of the F5 gene were subjected to PCR and direct sequencing. Suspected variant sites were confirmed by clone sequencing. Influence of the variants was predicted by using software including ClustalX and Mutation Taster. Results: The prothrombin time (PT) and activated partial thromboplastin time (APTT) of the proband were prolonged to 20.3 s and 59.2 s, respectively, while FⅤ activity (FⅤ∶C) and FⅤ antigen (FⅤ∶Ag) were reduced by 13% and 17%, respectively. The FⅤ∶C and FⅤ∶Ag of his father, sister and daughter were decreased to 35%, 37%, 29% and 42%, 46%, 35%, respectively. The proband was found to carry a heterozygous c. 2851delT variant in exon 13 of the F5 gene, which caused a frameshift and resulted in a truncated protein (p.Ser923LeufsX8). In addition, a heterozygous c. 1538G>A (p.Arg485Lys) variant was found in exon 10. The father, sister and daughter of the proband all carried the p. Ser923LeufsX8 variant, while his mother and son carried the heterozygous p. Arg485Lys polymorphism. His younger brother and wife were of the wild type. Bioinformatic analysis showed that p. Ser923 was highly conserved across various species. Mutation Taster scored 1.00 for the p. Ser923LeufsX8 variant, and the result has predicted a corresponding disease. Conclusion A heterozygous deletional mutation c. 2851delT in exon 13 of the F5 gene and a heterozygous c. 1538G>A polymorphism harbored by the proband may be associated with the decreased FⅤ level in this pedigree.

Objective To raise awareness of the late-onset meningitis caused by group B streptococcus (GBS) which was homogenous to the maternal colonization.Methods The clinical data of late-onset GBS meningitis in neonates twins whose pathogens were homogenous to their maternal colonization were collected from Department of Neonatology,Guangzhou Women and Children's Medical Center.The general conditions,clinical symptoms,laboratory tests and drug treatment of the twins and their mother were retrospectively analyzed,and the GBS homology during inpatient care was tested.And the progress of the twins' condition was investigated by telephone follow-up.Results The mother had two pregnancies without prenatal GBS screening or intrapartum antimicrobial intervention for GBS,everything was normal during pregnancy and delivery.Twins were born through cesarean section.The elder sister was discharged with Linezolid taken orally after 167 days in hospital without convulsions,shaking or other discomfort.The elder sister was followed up for every 2 weeks,and in the last time of follow-up,cerebrospinal fluid white blood cell counts were 45 × 106/L,protein level was 1.52 g/L and Linezolid was withdrawn.The younger brother was discharged after 58 days in hospital with follow-up for every 2 weeks,and in the last time of follow-up,cerebrospinal fluid white blood cell counts were 30 × 106/L,protein level was 0.66 g/L.During the hospitalization and follow-up without convulsions and irritation,and the cranial magnetic resonance imaging of the twin brother was normal.Test results showed that the GBS bacteria strain for twins and their mother were all serotype Ⅲ.The possibility of the GBS homology was more than 90％.Conclusions The toxicity of serotype Ⅲ GBS strain was strong.More proactive precautions should be considered to apply for the mother whose first birth already had GBS infection.Early identification and intervention of infection risk factors would help optimize the anti-infection treatment program and reduce nerve system damage and other adverse outcomes caused by invasive GBS infection.

OBJECTIVETo explore the molecular pathogenesis for a pedigree affected with coagulation factor Ⅴ (FⅤ) deficiency.

METHODSProthrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), coagulation factor Ⅱ activity (FⅡ: C), FⅤ activity (FⅤ: C), coagulation factor Ⅶ activity (FⅦ: C), and coagulation factor Ⅹ activity (FⅩ: C) were determined with a STAGO automatic coagulometer. FⅤ antigen (FⅤ: Ag) was detected with enzyme linked immunosorbent assay (ELISA). All exons and their flanking regions, and 5' and 3' untranslated regions of the F5 gene were analyzed by direct sequencing. Suspected mutation was verified by reverse sequencing as well as testing of family members. ClustalX software was used to analyze the conservative property of the mutation sites. PROVEAN and MutationTaster online software was used to predict the effect of the mutation on the protein function. Swiss-pdbViewer was used to analyze the protein model and interaction of amino acids.

RESULTSThe PT and APTT of the proband were slightly prolonged to 15.2 s and 41.8 s, respectively. And the FⅤ: C and FⅤ: Ag measured 55% and 62%, respectively. The FⅤ: C and FⅤ: Ag of his father and son were decreased to various extent (60%, 65% and 31%, 40%, respectively). A c.911G>A heterozygous mutation (Gly276Glu) was detected in exon 6 of the proband, for which her father and son were heterozygotes. The same mutation was not found in her mother, brother and husband. Conservation analysis showed that the Gly276 is highly conserved across various species. By bioinformatic analysis, the PROVEAN (scored -6.214) indicated Gly276Glu was harmful, and MutationTaster (scored 0.976) suggested that it is pathogenic. Model analysis suggested there are two hydrogen bonds between Gly276 and Ile298 in the wild type protein. When Gly276 was replaced by Glu276, the original hydrogen bond did not change, but the side chain of Glu was extended, which added steric hindrance with the surrounding amino acids, which resulted in decreased protein stability.

CONCLUSIONThe heterozygous c.911G>A (Gly276Glu) mutation of the F5 gene probably underlies the decreased level of FⅤin the proband.

Adult ; Computational Biology ; Factor V ; chemistry ; genetics ; Factor V Deficiency ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Pedigree ; Phenotype

Objective To prevent and treat of ceramic membrane purification of membrane fouling process of TCM extracts; To explore new methods of forecasting membrane fouling degree.Methods BP neural network model was improved. Methods to fast determine the optimal number of neurons in the hidden layer and fast algorithm for optimizing the weight and threshold of BP neural network were studied. Data of 207 groups of TCM extracts were under network training and prediction.ResultsCompared with the models of multiple regression analysis, basic BP neural network and RBF neural network, the error of the improved BP neural network model was less than that of the BP neural network model, and the mean square error was only 0.0057. In addition, the improved BP neural network model performance was more stable. In the 20 random running experiments, the goal of the success rate achieved up to 95%.Conclusion The improved model has a good network performance, the fitting effect and prediction ability, and can forecast the fouling degree of membrane stably and accurately.

Objective To investigate the correlation between the degree of liver fibrosis and Aspartate aminotransferase-to-Platelet Ratio Index ( APRI ) in children with biliary atresia ( BA ) , and evaluate the clinical significance of liver fibrosis in biliary atresia.Methods A total of 97 patients with diagnosed BA were recruited between January 2010 and June 2013.AST, PLT and APRI were determined one week before laparotomy.The severity of hepatic tibrosis was.Judged by Metavir system the correlation among AST, PLT, APRI and severity of liver fibrosis were evaluated, and their diagnostic value for degree of liver fibrosis was analyzed by ROC.Results Sera AST levels and PLT counts of BA patients were found to be positively(r=0.367, P<0.01) and negatively(r=-0.403, P<0.01) correlated with Metavir scores of liver fibrosis, respectively.There existed positive correlation between APRI and the severity of hepatic fibrosis (r=0.541, P<0.01).The area under ROC curve of APRI to diagnose none or mild fibrosis and moderately severe fibrosis was 0.78, with sensitivity of 77.9%and specificity of 62.1%at the optimal cut-off value of 0.75; the area under ROC curve of APRI to diagnose moderately severe fibrosis with liver cirrhosis arrived 0.85, with sensitivity of 75.0% and specificity of 89.4% at the optimal cut-off value of 1.77.The accuracy of none or mild fibrosis, moderate fibrosis and cirrhosis diagnosed by APRI were 73.2%, 64.9%, 87.6%, respectively.Conclusion APRI can be used as a non-invasive parameter to assess the severity of hepatic fibrosis with BA.

Gastrin is mainly secreted by the G cells in antrum and the upper part of small intestine.Gastrin receptor distributes in various tissues.Gastrin and its receptor have several functions including regulating cell growth and differentiation,inhibiting apoptosis,promoting cell proliferation,invasion and metastasis.Studies have shown that gastrin and its receptor involve with various cancers occurrence and development.Gastrin and its receptor can be used to diagnosis early gastric cancer,and be used as a potential targets in gastric carcinoma treatment.The relationship of gastrin and its receptor with gastric carcinoma were reviewed in this paper.