Objective To explore the effects of Neurogenesin 1 (Ng1) gene on functional recovery after spinal cord injury (SCI) and its mechanism. Methods Thirty-six rats (aging 4 months, weighing 230 g and being male or female), were randomly divided into two groups: experimental group (n=18) and control group (n=18). After spinal cord contusive injury at T10 level was made in all these rats using modified Allen’s method, Ng1 recombinant plasmid and blank plasmid were transfectedinto the damaged areas of exprimental group and control group respectively by Alzet pumps. At 1 day, 1 week, 2 weeks, 3 weeks, and 4 weeks after SCI, Basso-Beattle-Bresnahan (BBB) Rating Scale was used to observe the recovery of motor function. At 1 week after injury, the expressions of Ng1 mRNA and protein in injured spinal cord were detected by RT-PCR and Western blot techniques. And at 2 and 4 weeks, double immunofluorescence and histopathologic examinations were performed to study the prol iferation of the adult endogenous neural stem cells and pathological change after SCI. Results At 1-4 weeks after SCI, the BBB scores in the exprimental group was significantly higher than that in control group (P lt; 0.05), and at 4 weeks the BBB score of the experimental group (16.80 ± 1.79) was significantly higher than that of the control group (9.60 ± 1.67), (P lt; 0.01). RTPCR and Western blot showed that the mRNA and protein expressions of Ng1 were observed in the exprimental group and no expression was seen in the control group. Histologic observation showed that the morphology of spinal cord and neurons in the exprimental group was better than that in the control group and was close to the normal tissue. The mean number of Nestin+/ BrdU+ newborn endogenous neural stem cells in the exprimental group was significantly more than that in control group (P lt; 0.05). Conclusion Ng1 gene could promote the prol iferation of endogenous neural stem cells and protect the injured neurons, which enhances the repair of the motor function after SCI.
OBJECTIVE: To investigate the variation of neurotrophic factors expression in spinal cord and muscle after root avulsion of brachial plexus. METHODS: Forty-eight Wistar rats were involved in this study and according to the observing time in 1st day, 1st week, 4th week, 8th week, and 12th week after avulsion, and the control, were divided into 6 groups. By immunohistochemical and hybridization in situ assays, the expression of nerve growth factor (NGF) on muscle, basic fibroblast growth factor(bFGF) and its mRNA on the neurons of corresponding spinal cord was detected. Computer image analysis system was used to calculate the result. RESULTS: After the root avulsion of brachial plexus occurred, expression of NGF increased and reached to the peak at the 1st day. It subsided subsequently but was still higher than normal control until the 12th week. While expression of bFGF and its mRNA increased in the neurons of spinal cord and reached to the peak at the 1st week. Then it dropped down and at the 12th week it turned lower than normal control. CONCLUSION: After root avulsion of brachial plexus, neurotrophic factors expression increase on target muscle and neurons of corresponding spinal cord. It maybe the autoregulation and may protect neuron and improve nerve regeneration.
Purpose To investigate the characteristics of intraocular growth of mice embryonic stem cells (ESC) in nude mice. Methods The undifferentiated murine ESC in vitro were transplanted into the eyes of nude mice.Mophological and immunohistochemical examinations were implemented. Results Two to three days after transplantation,yellowish-white granules and masses were seen inside the anterior chamber and vitreous cavity and enlarged gradually.Morphological examination showed that there were undifferentiated cells and differentiated cells in anterior chamber and vitreous cavity.The morphology and alignment of some differentiated cells were similar to those of the retina of nude mice.The cells were highly positive in NSE staining. Conclusion The transplanted ESC could grow in the eyes of nude mice and differentiate into neurons and retina-like structure. (Chin J Ocul Fundus Dis，2000，16：213-284)
Objective To evaluate clinical significance of reversed sural neurovascular fasciocutaneous flap for reconstruction of softtissue defects in ankle and foot. Methods From July 1994 to December 2002, 52 cases of soft-tissuedefects in the ankle and foot were reconstructed by use of reversed sural neurovascular fascio-cutaneous flap, including 47 cases of traumatic defects, 3 cases of chronic ulcer and 2 cases of tumors. The flap area ranged from 4 cm×6 cm to 10 cm×21 cm. Results The flaps survived in 48 cases; the distal part necrosed and secondary free-skin graft were further conducted in 4 cases. All soft-tissue defects were repaired and their accompanied bone and tendon exposurehealed. Forty-six cases were followed-up for 5 months to 48 months, the color and texture of the flaps were excellent and 2point discrimination was 11-17 mm(14 mm on average).The functions of ankle joints were good.Conclusion The reversedsural neurovascular fascio-cutaneous flap is convenient in design and dissection. Its use can retained and replace vascular anostomosed flaps to certain degrees.
Objective To study the microstructural change of detrusor muscle and neuromuscular junction (NMJ) after bladder functional reconstruction for atonic bladder caused by medullary cone injury and to discuss the feasibility of bladder functional reconstruction for improving the detrusor muscle degeneration. Methods A total of 104 adult female Sprague-Dawley rats (weighing, 200-250 g) were randomized divided into 3 groups: normal group (n=8), control group (n=48), and experimental group (n=48). No treatment was given in normal group; the medullary cone injury was established by sharp transection of spinal cord at L4, 5 levels in control group; and the anastomosis of bilateral L5 ventral root (VR)-S2 VR and L5 dorsal root (DR)-S2 DR was performed for bladder functional reconstruction after modeling of medullary cone injury in experimental group. After operation, the survival condition of rats was observed. At 3 days and 3 consecutive days before 1, 2, 3, 4, 5, and 6 months after operation, the residual urine volume was measured; at 1, 2, 3, 4, 5, and 6 months after operation, the detrusor muscle was harvested to measure the muscle fiber cross-sectional area by HE staining, to calculate the percentage of connective tissue by Masson trichrome staining, and to observe the ultrastructure of the detrusor muscle and the NMJ by transmission electron microscope (TEM). Results Eleven rats were supplemented because of death after operation. In control group, a significant increase of the residual urine volume was observed with the extension of time (P lt; 0.05); in experimental group, an increase was observed at the first 3 months after operation, and then gradually decreased, showing significant differences between the other time point (P lt; 0.05) except between at 3 days and at 5 months after operation (P gt; 0.05); there was significant difference between control and experimental groups at other each time point (P lt; 0.05) except at 3 days, 1 month, and 2 months (P gt; 0.05). HE staining and Masson trichrome staining indicated that the muscle fibers arranged in disorder with gradually aggravated atrophy and gradually increased connective tissue in control group, while the shape of the detrusor muscle recovered with no increased connective tissue at 4, 5, and 6 months after operation in experimental group; there was significant difference in cross-sectional area of detrusor muscle and percentage of connective tissue between normal group and experimental group, and between normal group and control group at each time point (P lt; 0.05). In control group, the cross-sectional area of detrusor muscle decreased and the percentage of connective tissue increased with the extension of time (P lt; 0.05). In experimental group, the cross-sectional area of detrusor muscle decreased at the first 3 months and then increased, and the percentage of connective tissue increased slowly with the extension of time. There was no significant difference of cross-sectional area of detrusor muscle at the first 3 months between control and experimental groups (P gt; 0.05), but the values in experimental group were significantly higher than those in control group at 4, 5, and 6 months after operation (P lt; 0.05). There were significant differences of the percentage of connective tissue between control and experimental groups at each time point (P lt; 0.05). In control group, the amount of synaptic vesicles decreased in the NMJ with time passing; vacuole like structure was observed in NMJ at 3 months; there was almost no nerve ending at 6 months. In experimental group, the amount of synaptic vesicles decreased at 1 and 3 months after operation, but obviously increased at 6 months. Conclusion The reconstruction of bladder function with L5 nerve roots above the paraplegic plane can effectively inhibit the degeneration of detrusor muscle and improve its microstructural changes after medullary cone injury.
A controversy still exists in the management of nerve injection injury. The results of different timing of operation and methods in treating this type of nerve injury were analysed in limb s function, neuroelectrophysiology and histology. The results showed that the recovery of the injuried nerve in the group of operation, was considerably better than that in the group without operation. In the group of operation early incision of the epineurium with saline irrigation! was superior to late neurolysis. It was suggested that the early incision with saline irrigation could be used as an emergency management for this type of nerve injury.
Objective To explore the method that can inducethe mesenchymal stem cells (MSCs) to differentiate into the neuronlike cells in vitro．Methods The neuron-like cells were isolated froman SD rat (age, 3 months; weight, 200 g). They underwent a primary culture; theinduced liquid supernatant was collected, and was identified by the cell immunohistochemistry. The C3H1OT1/2 cells were cultured, as an MSCs model, and they were induced into differentiation by β-mercaptoethanol (Group A) and by the liquid supernatant of the neuron-like primary cells (Group B), respectively. The cells were cultured without any induction were used as a control (Group C). Immunohistochemistrywas used to identify the type of the cells． Results The result of the immunochemistry showed that the cells undergoing the primary culture expressed the neurofilament protein (NF) and the neuronspecific enolase (NSE), and they were neuron-like cells. β-mercaptoethanol could induce the C3H1OT1/2 cells toexpress NF and NSE at 2 h, and the expression intensity increased at 5 h. The liquid supernatant of the primarily-cultured neuron-like cells could induce theC3H1OT1/2 cells to express NF and NSE at 1 d, but the expression intensity induced by the liquid supernatant was weaker than that induced by β-mercaptoethanol. The positivity rate and the intensity expression of NSE were higher than those of NF. Conclusion MSCs can differentiate into the neuron-like cells by β-mercaptoethanol and the microenvironment humoral factor, which can pave the way for a further study of the differentiation of MSCs and the effectof the differentiation on the brain trauma repair.
Objective To investigate the possibility of commitment differentiation of embryonic stem cells induced by the medium of cultured retinal neurons of SD rats. Methods The medium from cultured retinal neurons of SD rats were collected, sterilized and mixed with DMEM medium according to 2∶3 proportion, ES cells were cultured with these mixed medium and were observed under the phase contrast microscope daily, the induced cells were identified by NF immunohistochemistry methods. Results The ES cells cultured with these mixed medium can differentiate into neuron-like structure, and the induced cells were positive in NF immunofluorescence staining. Conclusion The medium from cultured retinal neurons of SD rats can induce ES cells commitment differentiation into neuron-like structure. (Chin J Ocul Fundus Dis, 2002, 18: 134-136)
In this experiment, two proximal ends of themedian and ulnar nerves of rabbit wereapproxirnated within the chitin tube for thepurpose to inhibit the neuroma formation. Byobservation under light and transmission electronmicrnscopo and immunohistochemistry, wefound that: (1) the axons of the two proximalstumpe could regenerate in the chitin tube for 2to 5mm, and then ceased to grow when anaxonal overlap happened resulting in inhibitingneuroma formation; (2) chitin tube could bedegradated a...
Objective To summarize the application and advancement of liver transplantation for hepatic metastasis from neuroendocrine tumor. Methods Domestic and overseas publications on the study of liver transplantation for hepatic metastasis from neuroendocrine tumor in recent years were collected and reviewed. Results Liver transplantation can offer good relief of symptoms, long disease-free intervals, and potential cure in individual patients with hepatic metastatic tumor. Important selection criteria are well-differentiated tumors and a low proliferation rate (Ki67＜10%). Conclusion In carefully selected patients with metastatic neuroendocrine tumors, liver transplantation is an appropriate option.