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The alpha regulator subunit B'' of protein phosphatase 2 (PPP2R3A), a regulatory subunit of protein phosphatase 2A (PP2A), was reported to present a special subcellular localization in cardiomyocytes and elevate in non-ischemia failing hearts. PPP2R3A has two transcriptions PR72 and PR130. PR72 acts as a negative regulator of the Wnt signaling cascade, while the Wnt signaling cascade plays a pivotal role in cardiac development. And PR130 was found to be involved in cardiac development of zebrafish in our previous study. Thus, to investigate the function of PR72 in heart, two stable pr72 knockout (KO) zebrafish lines were generated using Transcription Activator-Like Effector Nuclease (TALEN) technology. Homozygous pr72 KO fish struggled to survive to adulthood and exhibited cardiac developmental defects, including enlarged ventricular chambers, reduced cardiomyocytes and decreased cardiac function. And the defective sarcomere ultrastructure that affected mitochondria, I bands, Z lines, and intercalated disks was also observed. Furthermore, the abnormal heart looping was detected in mutants which could be rescued by injection with wild type pr72 mRNA. Additionally, it was found that Wnt effectors were elevated in mutants. Those indicated that deletion of pr72 in zebrafish interrupted cardiac development, probably through activation of the Wnt pathway.

BACKGROUND The morbidity and mortality of lung cancer in Xuanwei (LCXW) is among the highest in China for both males and females and increases constantly. The underlying molecular changes associated with LCXW are still unclear. The purpose of this study was to screen out potential cancer-related genes which may become promising biomarkers of LCXW. METHODS Differentially expressed genes (DEGs) were detected by the expression microarrays in 29 paired LCXW tissues (tumor tissue and matched adjacent non-cancerous lung tissues). Integrated with bioinformatic analyses, a literature review was applied for screening out important cancer-related genes. An additional 44 paired LCXW samples were collected to verify the transcription and expression levels of the candidate gene by qRT-PCR and Western blotting. The relationship between the candidate genes and clinical pathological features were evaluated using Fisher's test. RESULTS mRNA expression microarrays revealed that the LCXW patients harbored 2,424 differentially expressed genes (fold change ≥ 2.0). Of these genes, 1,636 were up-regulated while the other 788 genes were down-regulated. Forty previously reported DNA repair genes associated with PAHs exposure were identified by microarrays. Bioinformatic analyses indicated down-regulated ANXA3 in LCXW patients which was closely related to pathological stage and involved in the regulation of a variety of biological responses. Review of the literature on ANXA3 found its down-regulation was a distinct expression pattern compared with lung cancer occurring in other geographic areas. qRT-PCR and Western blotting confirmed the down-regulation of ANXA3 was associated with LCXW. The correlation analysis of clinical features showed down-regulated ANXA3 was correlated to the progression of pathological stage. CONCLUSIONS The LCXW patients harbored more DEGs, and these DEGs were involved in a wide range of pathways and biological function damage. We preliminarily suggested ANXA3 may be a potential LCXW related gene. ANXA3 may serve as a promising biomarker for diagnosis of LCXW.

Background In lung cancer, miR-29b can act as a tumor suppressor, influencing epigenetic regulation, cell proliferation, differentiation, apoptosis, metastasis, and chemosensitivity. However, its expression and biological function in Xuanwei lung cancer (XWLC) remains unknown. Methods Fifty pairs of XWLC and matched normal tissues were collected and analyzed for the expression of miR29b by quantitative RT-PCR. MiR-29b precursor and inhibitor were transfected into XWLC-05 cells to investigate its role on regulating cell proliferation by MTT assay and cell apoptosis by caspase 3/7/9 assay in vitro. The target genes of miR-29b were predicted and determined by bioinformatics analysis and luciferase reporter assay. Results qRT-PCR results showed that the expression level of miR-29b in XWLC tissues was significantly lower than that in the matched normal tissues, with a proportion of 86%. The low expression of miR-29b was significantly associated with lymphatic metastasis. MiR-29b could suppress cell proliferation and promote apoptosis in XWLC-05 cells. According to the bioinformatics analysis and luciferase reporter assay results, PUMA was a target gene of miR-29b in XWLC. Conclusions Our studies provided evidence for the involvement of miR-29b in XWLC lymphatic metastasis and suggested that miR-29b may serve as a new biomarker for diagnosis of XWLC or therapeutic targets.

To analyze genomic copy number variations (CNVs) in two sisters with primary amenorrhea and hyperandrogenism.G-banding was performed for karyotype analysis. The whole genome of the two sisters were scanned and analyzed by array-based comparative genomic hybridization (array-CGH). The results were confirmed with real-time quantitative PCR (RT-qPCR).No abnormality was found by conventional G-banded chromosome analysis. Array-CGH has identified 11 identical CNVs from the sisters which, however, overlapped with CNVs reported by the Database of Genomic Variants (http://projects.tcag.ca/variation/). Therefore, they are likely to be benign. In addition, a -8.44 Mb 9p11.1-p13.1 duplication (38,561,587-47,002,387 bp, hg18) and a -80.9 kb 4q13.2 deletion (70,183,990-70,264,889 bp, hg18) were also detected in the elder and younger sister, respectively. The relationship between such CNVs and primary amenorrhea and hyperandrogenism was however uncertain. RT-qPCR results were in accordance with array-CGH.Two CNVs were detected in two sisters by array-CGH, for which further studies are needed to clarify their correlation with primary amenorrhea and hyperandrogenism.

We investigated the genetic defects in two families with classic congenital adrenal hyperplasia (CAH). Clinical data and vein blood of the family members were collected, hormonal evaluation, karyotype analysis, ultrasound and CT scans were performed, and a direct-sequencing of PCR products of the candidate genes was used to identify the mutations. In family A, Patients A-II:1 and A-II:2 were found to be in a compound heterozygous state for mutations of p.I172N (g.1004 T > A) in exon 4 and IVS2-13A/C > G (g.659A/C > G) in intron 2 in CYP21A2 gene, their father A-I:1 and mother A-I:2 were found to carry a heterozygous mutation of IVS2-13A/C > G (g.659A/C > G) and p.I172N (g.1004 T > A) respectively. In family B, Patient B-II:1 was detected to carry only one heterozygous mutation of p.I172N; no other mutations in CYP11B1, CYP17A1 or HSD3B2 genes were detected. Her father B-I:1 carried a heterozygous mutation of p.I172N (g.1004 T > A) and her mother B-I:2 was found to be a wild type in all the candidate genes. Obviously, patients A-II:1 and A-II:2 inherited the p.I172N (g.1004 T > A)-bearing maternal allele and the IVS2-13A/C > G (g.659A/C > G)-bearing paternal allele. And this kind of compound heterozygous mutations caused simple virilising (SV) CAH. While patient B-II:1, whose phenotype was SV CAH too, was found to carry only one heterozygous mutation of the p.I172N (g.1004 T > A)-bearing paternal allele, and needed further studies.

BACKGROUND Long noncoding RNAs (lncRNAs) play an important role in various biological processes involved the development and progression of lung cancer. However, their expression signature in Xuanwei lung cancer (XWLC) remains unknown. METHODS High throughput microarray assay was performed to detect lncRNA and mRNA expression profiles in five paired human XWLC and adjacent normal tissues. Bioinformatic analyses (gene ontology, pathway, and net-work analysis) were applied for further study of these differentially expressed mRNAs. An additional 33 paired XWLC samples were collected to verify the expression levels of 6 candidate lncRNAs using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). RESULTS Using abundant and varied probes, about 37,000 lncRNAs and 34,000 mRNAs were assessed in our micro-array. 1,484 lncRNAs and 1,997 mRNAs were differentially expressed in XWLC and adjacent normal samples (fold change ≥ 2.0). Using qRT-PCR validation, six candidate lncRNAs were differentially expressed in XWLC and adjacent normal tissues. The qRT-PCR results were consistent with the microarray data. Among these, FENDRR and SGOL1-AS1 were the most aberrantly expressed lncRNAs. CONCLUSIONS This study firstly ascertained the expression profile of lncRNAs in XWLC by microarray. The results revealed that many lncRNAs were differentially expressed in XWLC and adjacent normal tissues. Further investigation of the differentially expressed lncRNAs may serve as new biomarkers for diagnosis of XWLC or novel therapeutic targets.