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Liu B.,Key Laboratory of Freshwater Fish Reproduction and Development | Liu B.,University of South China | Ke J.,Chongqing Medical University | Cai S.,Chongqing University | And 8 more authors.
Shengwu Gongcheng Xuebao/Chinese Journal of Biotechnology | Year: 2012

A scaffold fabricated with lysine/nerve growth factor (NGF)/poly (lactic acid coglycolic acid) copolymer (PLGA) and acellular pigskin was evaluated in vitro as a potential artificial nerve scaffold. Properties of the scaffold such as microstructure, mechanical property, degradation behavior in PBS and water, Schwann cell adhesion property, and release of NGF were investigated. Results showed PLGA had permeated into the porous structure of acellular pigskin; its breaking strength was 8.308 MPa, breaking extensibility was 38.98%, elastic modulus was 97.27 MPa. The porosities of the scaffold ranged from 68.3% to 81.2% with densities from 0.62 g/cm3 to 0.68 g/cm3. At 4 weeks of degradation in vitro, maximum mass loss ratio was 43.3%. The release of NGF could still be detected on the 30th day, and its accumulative release rate was 38%. Lysine added into the scaffold neutralized the acidoid preventing degradation of PLGA to maintain a solution pH value. Schwann cells had grown across the scaffold after co-cultivation for 15 days. These in vitro properties of the pigskin based composite might indicate its potentiality to be an artificial nerve scaffold.

Zhang L.,Southwest University | Zhang L.,Key Laboratory of Freshwater Fish Reproduction and Development | Zhang J.,Chongqing Medical University | Li B.,Chongqing Medical University | And 3 more authors.
Shengwu Gongcheng Xuebao/Chinese Journal of Biotechnology | Year: 2014

Neural crest stem cells originated from hair follicle (epidermal neural crest stem cell, EPI-NCSC) are easy to obtain and have potentials to differentiate into various tissues, which make them eminent seed cells for tissue engineering. EPI-NCSC is now used to repair nerve injury, especially, the spinal cord injury. To investigate their effects on repairing peripheral nerve injury, EPI-NCSC from a GFP-SD rat were primarily cultured on coated dishes and on a poly lactic acid coglycolic acid copolymer (PLGA) membrane. Methyl thiazolyl tetrazolium (MTT) assay showed that the initial adhesion rate of EPI-NCSC was 89.7% on PLGA membrane, and the relative growth rates were 89.3%, 87.6%, 85.6%, and 96.6% on the 1st, 3rd, 5th, 7th day respectively. Cell cycles and DNA ploidy analysis demonstrated that cell cycles and proliferation indexes of cultured EPI-NCSC had the same variation pattern on coated dishes and PLGA membrane. Then cultured EPI-NCSC were mixed with equal amount of extracellular matrix and injected into a PLGA conduit to connect a 10 mm surgery excision gap of rat sciatic nerve, Dulbecco's Modified Eagle's medium (DMEM) was used to substitute EPI-NCSC in the control group. After four weeks of transplantation, the defected sciatic nerve achieved a histological restoration, the sensory function of rat hind limb was partly recovered and the sciatic nerve index was also improved. The above results showed that a PLGA conduit filled with EPI-NCSC has a good repair effect on the peripheral nerve injury. ©2014 Chin J Biotech. All rights reserved.

Lu H.,Ocean University of China | Lu H.,Southwest University | Lu H.,Key Laboratory of Freshwater Fish Reproduction and Development | Zhang X.,Ocean University of China | And 7 more authors.
Chinese Journal of Oceanology and Limnology | Year: 2014

The present study compared vertebral deformities of hatchery-reared and wild-caught juvenile Japanese flounder, Paralichthys olivaceus. A total of 362 hatchery-reared flounder (total length 122.5–155.8 mm) were collected from three commercial hatcheries located in Yantai, East China, and 89 wild fish (total length 124.7–161.3 mm) were caught off Yangma Island near Yantai City (37°27′N, 121°36′E). All the fish were dissected, photographed, and images of the axial skeleton were examined for vertebral deformities. Compared with wild-caught flounder in which no deformed vertebrae were detected, 48 (13.3%) hatcheryreared fish had deformed vertebrae. The deformities were classified as compression, compression-ankylosis, and dislocation-ankylosis. The vertebral deformities were mainly localized between post-cranial vertebra 1 and 3, with vertebrae number 1 as the most commonly deformed. The causative factors leading to vertebral deformities in reared Japanese flounder may be related to unfavorable temperature conditions, inflammation, damage, or rupture to the intervertebral ligaments under rearing conditions. Furthermore, no significant difference in the total number of vertebral bodies was observed between wild-caught (38.8±0.4) and hatchery-reared flounder (38.1±0.9) (P>0.05). However, the number of vertebral bodies of hatchery-reared and wild-caught flounder ranged from 35 to 39 and from 38 to 39, respectively. © 2015, Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag Berlin Heidelberg.

Huang S.,Key Laboratory of Freshwater Fish Reproduction and Development | Huang S.,Southwest University | Ma J.,Key Laboratory of Freshwater Fish Reproduction and Development | Ma J.,Southwest University | And 5 more authors.
Biochemical and Biophysical Research Communications | Year: 2011

Geminin plays an important role in coordinating the cell cycle with anterior-posterior patterning during embryonic development. However, whether it is involved in the regulation of left-right (LR) patterning remains unknown. Here, we reported that geminin is required for setting up heart and visceral laterality during zebrafish development. Defective heart and visceral laterality was observed in geminin morphants. Further study demonstrated that the left-sided nodal/. spaw in the lateral plate mesoderm (LPM) as well as the sideness of its downstream targets lefty2 and lefty1 was perturbed in geminin morphants. Upstream of the left-sided Nodal signal along the regulatory cascade of LR asymmetry, knock down of geminin resulted in defective Kupffer's vesicle (KV) formation and ciliogenesis rather than middle line defects. Predominant distribution of an antisense morpholino against geminin in dorsal forerunner cells (DFCs) led to defective KV morphogenesis and perturbed LR asymmetry, similar to those of geminin morphants, indicating a cell-autonomous role of geminin in regulating KV formation and ciliogenesis. Our results demonstrate that geminin is required for proper KV formation and ciliogenesis, thus playing an important part in setting up LR asymmetry. © 2011.

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