Erythropoietin inhibits apoptosis of retinal ganglion cells induced by high glucose through JNK signaling pathway.

The aim of this study was to investigate the effect of erythropoietin (EPO) on the apoptosis of retinal ganglion cells (RGCs) induced by high glucose and its mechanism. Rat primary RGCs were extracted to establish high glucose-induced apoptosis models using a 30 mM high-glucose medium. Then flow cytometry, cell counting kit-8 (CCK-8) assay and Western blotting assay were performed to detect the effects of high-, medium- and low-dose EPO on the apoptosis of RGCs induced by high glucose. Next, the molecular mechanism by which EPO suppressed the high glucose-induced apoptosis of RGCs was explored via gene array assay and bioinformatics analysis. The results and mechanism of bioinformatics analysis were verified by Western blotting assay. Finally, the small interfering ribonucleic acid (siRNA) experiment was applied to knock down tyrosine-protein phosphatase non-receptor type 1 (PTPN1) and PTPN11 to verify their roles in the inhibition of EPO on the apoptosis of RGCs triggered by high glucose. Flow cytometry-Annexin V/propidium iodide (PI) staining and CCK-8 assay confirmed that the high-, medium- and low-dose EPO inhibited the apoptosis of RGCs induced by high glucose in a dose-dependent manner (P<0.05). Subsequently, Western blotting assay results manifested that the high-, medium- and low-dose EPO reduced the expression levels of apoptosis-related proteins active-cysteinyl aspartate specific proteinase 3 (Caspase 3) and active- Caspase 9 in a dose-dependent manner (P<0.05). Moreover, according to gene array assay and bioinformatics analysis results, the c-Jun N-terminal kinase (JNK) signaling pathway, PTPN1 and PTPN11 might exert crucial effects in the inhibition of EPO on the apoptosis of RGCs induced by high glucose. Western blotting assay results also demonstrated that, compared with the high-glucose treatment, the high-dose EPO treatment decreased the protein expression level of phosphorylated (p)-JNK1/JNK but increased the protein expression levels of PTPN1 and PTPN11 (P<0.05). Moreover, flow cytometry-Annexin V/PI staining and CCK-8 assay results revealed that in EPO-treated cells, knocking down PTPN1 and PTPN11 significantly reversed the protective effect of EPO against high glucose-induced retinal ganglion cell apoptosis (P<0.05). Lastly, Western blotting assay illustrated that knocking down PTPN1 and PTPN11 significantly abolished the inhibition of high-dose EPO on the JNK signaling pathway. EPO may suppress the JNK signaling pathway by raising the expression levels of PTPN1 and PTPN11, so as to inhibit the apoptosis of RGCs triggered by high glucose.

Characterization of RsMYB28 and RsMYB29 transcription factor genes in radish (Raphanus sativus L.).

Glucosinolates (GSLs) are important secondary metabolites in Brassicaceae plants. Previous studies have mainly focused on GSL contents, types, and biosynthesis-related genes, but the molecular characterization patterns of GSL biosynthesis-related transcription factors remain largely unexplored in radish (Raphanus sativus L.). To isolate transcription factor genes regulating the GSL biosynthesis, genomic DNA and cDNA sequences of RsMYB28 and RsMYB29 genes were isolated in radish. Two R2R3-MYB domains were identified in the deduced amino acid sequences. Subcellular localization and yeast-one hybrid assays indicated that both the RsMYB28 and RsMYB29 genes were located in the nucleus and possessed transactivation activity. Reverse transcription quantitative analysis showed that the RsMYB28 and RsMYB29 genes were expressed in seeds, leaves, stems, and roots at the seedling, taproot thickening, and mature stages. Both genes were highly expressed during the seedling and taproot thickening stages. The expression level of RsMYB28 was found to be up-regulated following wounding, glucose, and abscisic acid treatments, whereas RsMYB29 was up-regulated following wounding and methyl jasmonate treatments. These results provide insights into the biological function and characterization of the RsMYB28 and RsMYB29 genes, and facilitate further dissection of the molecular regulatory mechanism underlying the GSL biosynthesis in radish.

Bispectral index for monitoring anesthetic depth in patients with severe burns receiving target-controlled infusion of remifentanil and propofol.

This study evaluated the feasibility and effectiveness of using the bispectral index (BIS) to monitor anesthetic depth in patients with severe burns receiving intravenous target-controlled infusion (TCI) of remifentanil and propofol. We randomly assigned 80 patients undergoing elective escharectomy (<1 week) to BIS (A) and control (B) groups. All patients received remifentanil and propofol as intravenous TCI anesthesia. Clinical data were recorded at different time points. The time from drug withdrawal to eye opening upon the patient hearing his/her name called and upon reaching an Aldrete score of 9 points was also recorded. During anesthesia maintenance, the target concentrations of remifentanil and propofol in group A were significantly lower than that in group B (2.12 ± 0.35 vs 2.50 ± 0.21 ng/mL and 2.54 ± 0.22 vs 2.86 ± 0.31 µg/mL, respectively; P < 0.01). The time from drug withdrawal to eye opening upon the patient hearing his/her name called and reaching an Aldrete score of 9 points in group A was considerably shorter than that in group B (7.90 ± 0.58 vs 8.35 ± 0.66 min and 9.15 ± 0.69 vs 11.13 ± 0.96 min, respectively; P < 0.01). In both groups, mean arterial pressure and heart rate values at each time point after loss of consciousness were significantly lower than the baseline values (P < 0.05), with the exception of 2 min after intubation. The use of BIS to monitor anesthetic depth in patients with severe burns receiving TCI of remifentanil and propofol during the perioperative period reduces propofol consumption and shortens the consciousness recovery time in patients.

Isolation and characterization of 24 polymorphic microsatellite loci in Sepioteuthis lessoniana.

Owing to their codominant, multiallelic, and highly polymorphic nature, microsatellite markers have been used widely in population genetics and biological resource conservation studies. To investigate the genetic structure of Sepioteuthis lessoniana, we developed 24 microsatellite DNA markers and assessed the polymorphism of each locus in a wild S. lessoniana population. The number of alleles per locus ranged from 4 to 26, and the observed and expected heterozygosities varied from 0.188 to 1.000 and 0.392 to 0.959 with an average of 0.675 and 0.852, respectively. These microsatellite loci will be useful tools in future studies of population genetic structure in this species.

[Bibliometric analysis on relations between cardiovascular disease and erectile dysfunction].

Objective: To understand the current situation and trend on the relations between erectile dysfunction (ED) and cardiovascular disease (CVD) through analyzing the epidemiologic research data. Methods: We conducted a literature search on the Scopus for potentially relevant epidemiologic studies on ED and CVD published from 1957 to October, 28, 2016. Age of the article, types, regions, citation, and co-authorship of the documents were recorded. Results: A total number of 412 pieces of literature were published in the past six decades, with original articles the most common types of ED and CVD. ED and CVD associated epidemiologic topics had an annual increase in number, and remained stable in the past decade, with occident countries as the United States and Italy taking the lead in this area. Clinical and epidemiological studies were the hottest areas, with most authors sharing a co-authorship. Conclusion: Our results suggested that inter-disciplinary cooperation with emphasize on clinical application were the effective starting points for ED and CVD associated epidemiologic studies.

First Report of Alternaria raphani Causing Black Patches on Chinese Radish During Postharvest Storage in Canada.

During November of 2003, Chinese radishes (Raphanus sativus cv. Taibai) harvested in St. Catharines, Ontario and stored in less than 1°C with 98% relative humidity (RH) and 5°C with 96% RH showed symptoms of black and dark brown, irregular patches, with or without decay. The symptoms were closely associated with skin wounds and damaged root hairs. Fungal DNA was extracted from discolored skin samples peeled from a radish, and 18S rRNA genes were amplified with fungal-specific PCR primers (1) EF4f (5'-ggaagggrtgtatttattag-3') and EF3r (5'-tcctctaaatgaccagtttg-3'). The cloned genes were sequenced using the primer EF4f and compared directly with nonredundant nucleotides in GenBank with BLAST. The results indicated that more than 75% of the fungal microflora on the diseased radish were Alternaria spp. Alternaria sp. was successfully isolated from discolored and decayed radish tissues. Morphological and molecular identification indicated that the isolated Alternaria sp. cultures belong to A. raphani, which was previously reported to cause leaf and pod blight on radish (2). For pathogenicity studies, a spore suspension (1 × 105 conidia/ml) obtained from a 4-week-old A. raphani culture was used to inoculate 'Taibai' Chinese radish tissues, including inner tissues and wounded and nonwounded skin. All tests were carried out at room temperature (22 to 24°C). On inner tissue and wounded skin, symptoms of dark brown-to-black patches appeared 2 days after inoculation and progressed with time. No symptoms developed on the noninoculated control or the nonwounded, inoculated treatment. A. raphani was reisolated from symptomatic tissue. Further evidence of pathogenicity was obtained by an additional inoculation and observation of symptoms. The results indicated that A. raphani was the causal agent of the black patches observed on Chinese radish, and to our knowledge, this is the first report that A. raphani could cause a postharvest disease on Chinese radish in storage. References: (1) J. D. Van Elsas et al. J. Microbiol. Methods 43:133, 2000. (2) M. S. Sangwan et al. J. Mycol. Plant Pathol. 32:125, 2002.