[Long term results of central hole type posterior chamber intraocular lens in the correction of moderate to high myopia].

Objective: To evaluate the long-term safety,effectiveness,predictability and stability of ICL V4c implantation for moderate to high myopia. Methods: In this retrospective case series study, 95 eyes from 50 patients with moderate to severe myopia who were treated in 2015 underwent central hole type posterior chamber intraocular lens (ICL V4c) implantation at Eye & ENT Hospital of Fudan University. The patients were followed up for a period of five years, during which we assessed various parameters including uncorrected visual acuity (UDVA), corrected visual acuity (CDVA), refractive error, axial length, intraocular pressure, endothelial cell density (ECD), vault, and complications. We used the paired t-test and repeated measures one-way ANOVA in SPSS statistical software to analyze the data. Results: The mean spherical equivalent refraction (SE) decreased significantly from (-12.16±3.04) D preoperatively to (-0.19±0.55) D at one month and (-1.14±0.84) D at five years postoperatively. The safety indices (postoperative CDVA/preoperative CDVA) were 1.24±0.27 and 1.13±0.27, respectively, and the efficacy indices (postoperative UDVA/preoperative CDVA) were 1.14±0.25 and 0.87±0.26 at one month and five years postoperatively. At one month after surgery, 80.00% of the eyes were within ±0.50 D of the expected correction, and 96.84% were within ±1.00 D. There was no significant difference in IOP between preoperative and postoperative measurements. The rate of ECD was 3.87%, and the vault decreased by 106.32 µm at five years postoperatively. Conclusion: ICL V4c implantation is safe and effective with good predictability and stability for long term.

Long non-coding RNA LINC00152 is up-regulated in ovarian cancer tissues and regulates proliferation and cell cycle of SKOV3 cells.

OBJECTIVE: To characterize functions of long non-coding RNA (lncRNA) in the progression of epithelial ovarian cancer. PATIENTS AND METHODS: Epithelial ovarian cancer tissues and matching normal tissues were collected from two individual patients for RNA microarray analysis. Besides, twenty-two ovarian cancer samples and ten healthy ovarian epithelial tissues were collected for Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR). Microarray assay suggested that a list of cancer relating mRNAs and lncRNAs were upregulated. The identified lncRNAs were validated via RT-qPCR, which led to the identification of long intergenic non-protein coding RNA 152 (LINC00152). To determine the function of LINC00152 in ovarian cancer, we knocked down the expression of LINC00152 in epithelial ovarian cancer cell line SKOV3 with small interference RNAs (siRNAs). The effects of LIN00152 on the proliferation and cell cycle were determined by comparing the cell viability of SKOV3 cells with LIN00152 knockdown and the control cells with negative siRNA. The cell viability was assessed using Cell Counting Kit-8 (CCK-8) and flow cytometry assay. RNA microarray assay was used again in control and LINC00152 knockdown SKOV3 cells to identify downstream signaling pathways. RESULTS: Fourteen ovarian cancer relating lncRNAs were identified by RNA microarray assay. Up-regulation of LINC00152 was validated via RT-qPCR. A higher expression of LINC00152 in late cancer stage (III-IV) compared to the early stage tumors was also demonstrated. Inhibition of LINC00152 in SKOV3 cells inhibited cell proliferation and induced cell cycle arrest that involved prolonged G1 phase and shortened S phase. The microarray assay data of SKOV3 cells suggested that Cyclin-Dependent Kinase Inhibitor 1C (CDKN1C) was a potential downstream target of LINC00152. CONCLUSIONS: LINC00152 is upregulated in epithelial ovarian cancer tissues comparing to normal tissues. Knockdown of LINC00152 expression inhibits cell proliferation and induces cell cycle arrest. LINC00152 possibly interacts with Tumor Necrosis Factor (TNF) signaling pathway. CDKN1C is a potential downstream target of LINC00152.

Effects of miR-21 on hypertensive rats through PTEN/PI3K/Akt/mTOR signaling pathway.

OBJECTIVE: The aim of this study was to investigate the effects of micro-ribonucleic acid (miR)-21 on hypertensive rats through the phosphatase and tensin homolog deleted on chromosome ten (PTEN)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway. MATERIALS AND METHODS: A total of 10 spontaneously hypertensive rats (SHRs) were selected as the model group. Meanwhile, 10 rats with the same age were enrolled in the normal control group. Real Time-fluorescence quantitative Polymerase Chain Reaction (qRT-PCR) was performed to detect the mRNA level of miR-21 in rats of the SHR model group and control group. The tail arterial diastolic pressure of rats in the awake and resting state was measured in both groups, respectively. Pathological sections were prepared to evaluate pathological changes in myocardial tissues. Subsequently, myocardial cells were isolated, cultured and transfected with miR-21 mimics and miR-21 inhibitor, respectively. Transfection efficiency was verified using fluorescence quantitative PCR. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was utilized to determine the apoptosis level of myocardial cells. Furthermore, the expression levels of the signaling pathway-related proteins were detected via Western blotting assay. RESULTS: Fluorescence quantitative PCR results revealed that the expression level of miR-21 was significantly higher in the SHR model group (p<0.05). The diastolic pressure increased markedly in the SHR model group when compared with that in the control group (p<0.05). Subsequent hematoxylin and eosin (HE) staining indicated apparent myocardial tissue injury in the SHR model group (p<0.05). After transfection, the results showed that miR-21 inhibitor could effectively down-regulate the expression level of miR-21 in myocardial cells (p<0.05). Meanwhile, TUNEL staining revealed that the number of apoptotic cells in the miR-21 inhibitor group was remarkably higher than that of the other two groups (p<0.05). In addition, Western blotting results manifested that the protein expression levels of PTEN, PI3K, Akt and mTOR were significantly lower in the miR-21 mimics group (p<0.05), whereas was remarkably higher in the miR-21 inhibitor group (p<0.05). CONCLUSIONS: MiR-21 is involved in regulating the pathological symptoms and myocardial cell apoptosis in hypertensive rats through the PTEN/PI3K/Akt/mTOR signaling pathway.

Characterization, culture medium optimization and antioxidant activity of an endophytic vitexin-producing fungus Dichotomopilus funicola Y3 from pigeon pea [Cajanus cajan (L.) Millsp.].

AIMS: The aim of this study was to characterize a fungal endophyte Y3 from pigeon pea (Cajanus cajan [L.] Millsp), as a novel producer of vitexin, and its culture medium optimization and antioxidant activity. METHODS AND RESULTS: The endophyte from the leaves of pigeon pea was identified as Dichotomopilus funicola by the morphological and molecular characteristics. The most important medium variables affecting vitexin production in liquid culture of D. funicola Y3 were screened by Plackett-Burman design, and three culture medium constituents (i.e. l-phenylalanine, salicylic acid and CuSO4 ·5H2 O) were identified to play significant roles in vitexin production. The most significant factors were further optimized using by central composite design with response surface methodology. The DPPH radical-scavenging assay indicated that fungal vitexin exhibited notable antioxidant activity with an EC50 value of 164 µg l-1 . CONCLUSIONS: First, a novel endophyte vitexin-producing Dichotomopilus funicola Y3 was isolated from pigeon pea (Cajanus cajan[L.] Millsp.). The maximum vitexin yield was obtained as 78·86 mg l-1 under the optimum culture medium constituents: 0·06 g l-1  l-phenylalanine, 0·21 g l-1 salicylic acid, and 0·19 g l-1 CuSO4 ·5H2 O in medium, which is 4·59-fold higher than that in the unoptimized medium. Also, fungal vitexin clearly demonstrated its antioxidant potential. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings provide an alternative source for large-scale production of vitexin by endophytic fungal fermentation and have a promising prospect in food and pharmaceutical industry.

New insights into mitogenomic phylogeny and copy number in eight indigenous sheep populations based on the ATP synthase and cytochrome c oxidase genes.

The origins and phylogeny of different sheep breeds has been widely studied using polymorphisms within the mitochondrial hypervariable region. However, little is known about the mitochondrial DNA (mtDNA) content and phylogeny based on mtDNA protein-coding genes. In this study, we assessed the phylogeny and copy number of the mtDNA in eight indigenous (population size, n=184) and three introduced (n=66) sheep breeds in China based on five mitochondrial coding genes (COX1, COX2, ATP8, ATP6 and COX3). The mean haplotype and nucleotide diversities were 0.944 and 0.00322, respectively. We identified a correlation between the lineages distribution and the genetic distance, whereby Valley-type Tibetan sheep had a closer genetic relationship with introduced breeds (Dorper, Poll Dorset and Suffolk) than with other indigenous breeds. Similarly, the Median-joining profile of haplotypes revealed the distribution of clusters according to genetic differences. Moreover, copy number analysis based on the five mitochondrial coding genes was affected by the genetic distance combining with genetic phylogeny; we also identified obvious non-synonymous mutations in ATP6 between the different levels of copy number expressions. These results imply that differences in mitogenomic compositions resulting from geographical separation lead to differences in mitochondrial function.

The ribosomal protein L34 gene from the mosquito, Aedes albopictus: exon-intron organization, copy number, and potential regulatory elements.

We describe the structural analysis of genomic DNA encoding ribosomal protein (rp) L34 from the mosquito, Aedes albopictus. Comparison of genomic DNA sequences encompassing approximately 8 kb with the rpL34 cDNA sequence showed that the gene contains three exons and two introns, encoding a primary transcript with a deduced size of 6196 nucleotides from the transcription start site to the polyadenylation site. Exon 1, which is not translated, measures only 45 bp, and is separated from Exon 2 by a 359 bp intron. Exon 2 measures 78 bp, and contains the AUG translation initiation codon 14 nucleotides downstream of its 5'-end. Downstream of Exon 2 is a 5270 bp intron, followed by the remainder of the coding sequence in Exon 3, which measures 444 bp including the polyadenylation signal. We used a novel PCR-based procedure to obtain 1.7 kb of DNA upstream of the rpL34 gene. Like the previously described Ae. albopictus rpL8 gene and various mammalian rp genes, the DNA immediately upstream of the rpL34 gene lacks the TATA box, and the rpL34 transcription initiation site is embedded in a characteristic polypyrimidine tract. The 5'-flanking DNA contained a number of cis-acting elements that potentially interact with transcription factors characterized by basic domains, zinc-coordinating DNA binding domains, helix-turn-helix motifs, and beta scaffold factors with minor groove contacts. Particularly striking was the conservation of an AP-4 binding site within 100 nucleotides upstream of the transcription initiation site in both Aal-rpL34 and Aal-rpL8 genes. Comparison of Southern hybridization signals using probes from the 5' and 3'-ends of the 5.3 kb second intron and the cDNA suggested that the Ae. albopictus rpL34 gene most likely occurs as a single expressed copy per haploid genome with restriction enzyme polymorphisms in the upstream flanking DNA and the likely presence of one or more pseudogenes.