Retinoic acid promotes formation of chicken (Gallus gallus) spermatogonial stem cells by regulating the ECM-receptor interaction signaling pathway.

Spermatogonial stem cells (SSCs) are the basis of spermatogenesis. Systematically exploring the critical factors associated with the formation of SSCs will provide new insight to improve the formation efficiency, and their practical application. Here we explore the regulatory mechanism of the ECM-receptor interaction signaling pathway and related genes during differentiation of SSCs in chicken. Firstly, the positive cell rate of SSCs protein marker was detected by immunofluorescence and flow cytometry and qRT-PCR was used to identify, the expression of related marker genes after 10 days of RA-induction. Secondly, the ESCs on 0d/ 4d /10d after RA- induction/self-differentiation were collected, and the total RNA was then extracted from cells. Finally, high-throughput analysis methods (RNA-seq) were used to sequence the transcriptome of these cells. After PCA analysis of the RNA-seq data, Venny analysis, GO and KEGG enrichment were further used to find the key signaling pathways and genes in the RA-induction process. The results showed that on day 10 of RA-induction, grape cluster growth cells expressed integrinß1, the specific marker protein of SSCs cells, and the integrinß1 positive rate was 35.1%. Also, SSCs marker genes CVH, Integrinß1, Integrinα6 were significantly up-regulated during RA-induction. Moreover, the significantly enriched pathway, ECM-receptor interaction signaling, in current study may play a crucial role in RA-induction. Then, JASPAR was used to predict the differential gene transcription factors in the signaling pathway, finding that RA receptor was a transcription factor of COL5A1, COL5A2 and COL3A1. The qRT-PCR results showed that the expression levels of RA receptors (RXRA, RARA and RXRG) and the predicted genes (COL5A1, COL5A2 and COL3A1) were both significantly increased during RA-induction. Also, dual-luciferase reporter assay showed that RA could affect the luciferin activities of COL5A1, COL5A2 and COL3A1. These results suggest that RA plays a crucial role in the formation of chicken spermatogonial stem cells via the transcription levels of COL5A1, COL5A2 and COL3A1 to regulate the ECM-receptor interaction signaling pathway. Additionally, knockdown of COL5A1/COL5A2/COL3A1 could effectively reduce the formation efficiency of SSCs. This indicated that the interference of RA receptor binding genes in the ECM-receptor interaction signaling pathway could decrease the efficiency of RA induced SSCs formation. Therefore, this study concludes that RA promotes formation of chicken spermatogonial stem cells by regulating the ECM-receptor interaction signaling pathway.

Treatment of stiff thoracic scoliosis by thoracoscopic anterior release combined with posterior instrumentation and fusion.

BACKGROUND: Thoracoscopic anterior release has been shown that it can effectively improve spinal flexibility in animal and human cadaveric studies, and has been advocated for use in patients with scoliosis. This prospective case series aims to investigate the improvement of the spinal flexibility and the effectiveness in deformity correction by anterior thoracoscopic release and posterior spinal fusion. METHODS: Eleven patients with stiff idiopathic thoracic scoliosis underwent anterior thoracoscopic release followed by posterior instrumentation. The average number of discs excised was five. Spinal flexibility was assessed by the fulcrum bending technique. Cobb angle before and after the anterior release was compared. RESULTS: The patients were followed for an average of 5.6 years (range 2.2 to 8.1 years). Fulcrum bending flexibility was increased from 39% before the thoracoscopic anterior spinal release to 54% after the release. The average Cobb angle before anterior release was 74 degrees on the standing radiograph and 45 degrees with the fulcrum-bending radiograph. This reduced to 34 degrees on the fulcrum-bending radiograph after the release, and highly corresponded to the 31 degrees measured at the post-operative standing radiograph. CONCLUSION: It was demonstrated in patients with stiff idiopathic thoracic scoliosis that thoracoscopic anterior spinal release can effectively improve the spinal flexibility and increase the correction of the spinal deformity.

[Surgical treatment of kyphosis with universal spine system].

OBJECTIVE: To introduce a method to treat kyphosis with universal spine system (USS) and to evaluate its clinical effect. METHODS: From January 2000 to September 2002, 31 patients with kyphosis were treated with "one-stage total vertebral wedge-shaped osteotomy" and internal fixation of USS. The average follow-up time was 12 months (6-31 months). We observed the clinical effect with the spinal column Cobb before operation and after operation, and the osteotomy confluence rate. RESULTS: In 31 patients, the average angle (Cobb) was 89 degrees before operation and 74 degrees after operation; the average rate of correction was 83.15% and the average rate of osteotomy confluence was 96.77%. No screw and rod broke and loosened. CONCLUSION: The USS is the reliable internal fixation instrument for the treatment of kyphosis, can increase the osteotomy confluence rate and improve correcting rate.