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Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⺠subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.
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Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⺠subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.
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Subject(s)
Buffaloes , Cells, Cultured , In Vitro Techniques , Primary Cell Culture , Stem Cells , TretinoinABSTRACT
Objective To compare characteristics of stress variations in 3D finite element models of normal and degenerative lumbar vertebrae and the dose-effect relationship, and analyze the mechanism of mechanical balance by traditional Chinese medicine (TCM) manipulation on degenerative lumbar vertebrae. Methods The 3D finite element model of intact, real human degenerative lumbar vertebrae (L4-5) was established to simulate the physiological activity of flexion and extension in lumbar vertebrae. The characteristics of stress variation in degenerative lumbar vertebrae under external loading, namely, the TCM manipulation was analyzed, and the stress variation in degenerative lumbar vertebrae under gradual increasing-external loading was analyzed as well, which was compared with the stress and strain variation in normal lumbar vertebrae under different motion status. Results Under different motion status, the stress distributions on lumbar disc as well as the elastic modulus of nucleus pulposus and fiber ring showed a gradually increasing tendency with lumbar degeneration increasing. TCM manipulation could change the stress distributions on lumbar disc, enlarge the space of spinal canal to a certain degree, and decrease the stress on nerve root. Stresses on small joints of the vertebral body and vertebral pedicle under posterior extension were larger than those under anterior flexion, while stresses on intervertebral disc under anterior flexion were greater than those under posterior extension, which showed a gradually increasing trend from top to bottom. Conclusions The mechanical environment of human lumbar vertebrae can be balanced by TCM manipulation, for the purpose of improving and treating lumbar disc diseases. The comparison with the 3D finite element model of normal human lumbar vertebrae and investigation on lumbar degeneration from perspective of changes in biomechanical environment and characteristics can provide scientific basis for clinic application of TCM manipulation in prevention and treatment of lumbar degenerative diseases, as well as new research idea for studying mechanical mechanism of TCM manipulation in effective prevention and treatment of lumbar lesions.
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BACKGROUND:Studies have shown that pulsed electromagnetism has a good effect in promoting peripheral nerve regeneration after cerebral infarction and spinal cord injury, and improving memory function in patients with neurological disorders. OBJECTIVE: To investigate the impact of pulsed magnetic field on brain function and endogenous neural stem cell factor in the brain tissue of rats with brain injury. METHODS: Totally 320 adult male Sprague-Dawley rats were randomly divided into model group, pulsed magnetic field 0.1 mT group, pulsed magnetic field 0.3 mT group and pulsed magnetic field 0.5 mT group (n=80 per group). After brain injury models were established using lateral hydraulic strike method, rats in the latter three groups were exposed to pulsed magnetic fields 0.1, 0.3, 0.5 mT, respectively. After electromagnetic radiation 1, 3, 7, 14 days, the motor function of rats was evaluated by beam-walking test and water maze test. Rats were intraperitoneally injected 5-deoxy-uridine (BrdU) at 1 day prior to different radiation time points, and BrdU and nestin expressions in the cerebral cortex were measured by immunohistochemical method. RESULTS AND CONCLUSION: (1) The time of water maze test and the beam-walking test at 1, 3 and 7 days after irradiation was ranked as follows: pulse magnetic field 0.5 mT < pulse magnetic field 0.3 mT < pulse magnetic field 0.1 mT < model group, and there were significant differences between groups (P < 0.05). (2) The expressions of BrdU and nestin at 1, 3 and 7 days after irradiation were highest in the pulse magnetic field 0.5 mT group, successively followed by pulse magnetic field 0.3 mT group, pulse magnetic field 0.1 mT group and model group (P < 0.05). In summary, the pulse magnetic field exhibits remarkable protective effects on the brain function of rats with craniocerebral injury in an intensity-dependent manner. The possible mechanism is related to the activation of neural stem cells and the proliferation of neural stem cells in the brain tissue of rats with craniocerebral injury.
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To investigate the epidemiological and clinical characteristics of emerging infectious gnathostomiasis in China,a retrospective analysis was performed on 16 cases of gnathostomiasis found during 2009-2011.Patients consisted of 11 women and 5 men.The median age was 48 years (ranging from 27 to 68years).Eleven out of 16 patients (68.8％) had history of ingesting raw or undercooked freshwater fish.The main manifestation of patients was cutaneous form.Clinical symptoms included migratory swelling,creeping eruption and skin rash.Among them,43.8％ (7/16) of patients presented eosinophilia.The results of serological tests showed all patients were Gnathostoma-specific IgG positive by dot immunogold fitration assay (DIGFA) and 5 cases (31.2 ％) were positive by immunoblot for detection of specific 24-kDa protein band reaction.The median time from onset of symptoms to diagnosis was 12 months with a range of 2 weeks to 3 years.A total of 11 patients were treated with albendazole and/or ivermectin.Ten patients did not relapse post treatment.During 2009-2011,the number of patients with gnathostomiasis significantly increased due to consumption of raw or under-cooked loaches and swamp eels.Serological test was a necessary tool to support clinical diagnosis while gnathostomiasis commonly presented complex symptoms.Ivermectin and albendazole were effective to treat gnathostomiasis.
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BACKGROUND: Nerve tissue-engineered scaffolds must have axial y aligned structures, that can promote oriented growth of new axons, to guarantee the effective repair and regeneration of damaged nerves. OBJECTIVE: To investigate the effect of heparin sulfate/col agen nerve tissue-engineered scaffolds on peripheral nerve injury repair. METHODS: Heparin sulfate/col agen nerve tissue-engineered scaffolds were prepared, and its internal structure and porosity was observed and measured. Then rat Schwann cel s were seeded on the scaffolds to observe cel adhesion. Afterwards, 32 rats undergoing removal of left sciatic nerve were randomly divided into two groups (n=16 per group), and the rats were implanted by heparin sulfate/col agen nerve tissue-ergineered scaffolds as experimentd group, and the rats were implanted by autdogous sciatic nerve as control group. At 16 weeks after implantation, diameter, thickness of myelin sheath as wel as density of myelinated nerve fiber, the percentage of neural tissue and electrophysiology was detected, respectively. RESULTS AND CONCLUSION: The tissue-engineered scaffolds whose porosity was 91% were composed of microtubules arranging paral el y along the axial direction, and the microtubule diameter was 180 μm; the scaffolds had good biocompatibility with the Schwann cel s. In addition, at 16 weeks after implantation, no significant differences were found in myelin sheath thickness, myelinated nerve fiber density, as wel as conduction velocity and latency of motor and sensory nerves between two groups;compared with the control group, diameter of myelinated nerve fiber, percentage of neural tissue and amplitude of motor and sensory nerves in the experimental group were significantly decreased (P < 0.05). To conclude, the heparin sulfate/col agen nerve tissue-engineered scaffold can effectively repair peripheral nerve injury, but its effect is weaker than that of autologous nerve repair.