Sperm cryopreservation in different grouper subspecies and application in interspecific hybridization.

The present study established and confirmed an efficient technology for groupers: giant grouper Epinephelus lanceolatus, orange-spotted grouper E. coioides, seven-band grouper, E. septemfasciatus, and kelp grouper E. moara sperm cryopreservation and successfully applied the cryopreserved E. lanceolatus sperm into interspecific hybridization with E. coioides. For both E. lanceolatus and E. coioides, the best motility of postthaw sperm were achieved using 6% to 10% DMSO, 6% to 16% propylene glycol, and 6% ethylene glycol as cryoprotectants. Furthermore, we have successfully applied this method into the other two species of E. septemfasciatus (74.56 ± 5.45%) and E. moara (71.67 ± 5.10%) sperm cryopreservation and obtained high motility, respectively. Computer-assisted sperm motion analysis showed that the postthaw sperm of the four species of grouper could keep 30 to 35 minutes motile state in nature seawater. And the freezing-thawing process decreased the sperm motility, speed, and longevity but did not significantly change the sperm movement pattern, and the progressive linear motion still was the dominant movement pattern. For the four species of grouper, the ultrastructural analysis showed 70% to 80% of the spermatozoa had intact morphology with a little of swelling; 20% to 30% were damaged, such as swelling or rupture of head, midpiece, and tail region; and 10% to 20% were severely damaged. Whereas, by the microscopic observation, more than 90% of the postthaw sperm presented normal morphology. In the artificial insemination and hybridization experiment, high fertilization rates and hatching rates were achieved when using 10% DMSO (88.7 ± 5.3%, 85.3 ± 7.4%) and 10% propylene glycol (86.8 ± 3.3%, 83.1 ± 6.6%), with no significant difference in comparison with control (92.2 ± 1.4%, 87.9 ± 4.2%). In addition, we found the embryos from postthaw sperm of E. lanceolatus and E. coioides eggs developed and grew normally as reported in previous study on hybridization of groupers (E. coioides × E. lanceolatus) using cryopreserved sperm. The results of the present study further validated the safety of the cryopreserved sperm in breeding production by assessing the fertilization capacity, embryo development, and larval growth.

Doublets versus single-agent therapy as first-line therapy for elderly patients with advanced non-small cell lung cancer? A systematic review of randomised controlled trials.

INTRODUCTION: Although the standard treatment for advanced non-small cell lung cancer (NSCLC) patients is platinum-based doublet chemotherapy, single-agent therapy is still preferred in elderly patients. Comparison of the efficacy of various combinations of doublets with single-agent chemotherapy is somehow contradictory. This study conducted a systematic review to evaluate the efficacy and tolerability of the third-generation agent-based doublets vs. single-agent chemotherapy in elderly NSCLC patients. METHODS: Electronic (PubMed, EMBASE and Cochrane Library database) and manual searches were conducted to collect data from published, randomised, phase 2 and 3 trials which compared doublets with a third-generation single-agent chemotherapy in elderly patients. Pooled relative risks (RRs) were calculated for the incidences of overall response rate (ORR), 1-year survival rate (1-y SR), and grade 3/4 toxicities. RESULTS: Seven eligible trials (2219 patients) were selected from 1170 studies that were initially identified. A significant difference in ORR favouring doublets over single agents was observed [RR, 1.59; 95% confidence interval (95% CI), 1.36-1.86; p < 0.0001] with a slightly, but not significantly improved 1-y SR (RR, 1.19; 95% CI, 0.98-1.45, p = 0.007). Subgroup analysis suggested that platinum (RR, 1.94; 95% CI, 1.47-2.55, p < 0.0001) or non-platinum- (RR, 1.45; 95% CI, 1.20-1.75, p < 0.0001) based doublets could improve ORR, and the grade 3/4 thrombocytopaenia (RR, 6.64; 95% CI, 1.78-24.86, p = 0.005) and anaemia (RR, 2.86; 95% CI, 1.62-5.05, p < 0.0001) were preferred to occur in platinum-based doublets. CONCLUSIONS: Doublets appear to be more effective and tolerable than single-agent therapy for treating elderly advanced NSCLC patients, and therefore could be considered as a treatment option for elderly populations with good physical status.

Germline-specific and sexually dimorphic expression of a dead end gene homologue in turbot (Scophthalmus maximus).

Germ cells are indispensable for gonadal development and fertility. However, the physiological mechanisms regulating germ cell development in marine fish are poorly understood due to a lack of germ cell markers. The dead end (dnd) gene is a vertebrate-specific component of germplasm crucial for primordial germ cells (PGCs) migration and development in teleosts. In this study, we identified a dnd homologue (Smdnd) in turbot (Scophthalmus maximus) and investigated its expression pattern during embryogenesis and gonadal development. The deduced amino acid sequence of Smdnd shared several conserved motifs of Dnd homologues as well as high identity to other Dnd proteins. Phylogenetic analysis revealed that the SmDnd was closely related to its teleost counterparts. Reverse transcription polymerase chain reaction (RT-PCR) and in situ hybridization revealed that Smdnd transcripts could be exclusively detected in germ cells, including presumptive PGC and adult male and female germ cells. In addition, an interesting sexually dimorphic expression of Smdnd during gonadal development was observed by real-time PCR. Female turbot showed greater (P < 0.05) Smdnd expression than male before sex maturation. This difference reduced gradually due to the upregulation of Smdnd in the male during the period corresponding to spermatogonia proliferation and meiosis. These results indicate that Smdnd can be used as a germ cell marker in turbot. In addition, the temporal and sex differences in Smdnd expression indicate that this gene may play different roles in gonadal development in both sexes.