Objective To observe whether Nogo-66 can inhibit the neurite outgrowth during the neuronal differentiation of the neural stem cells (NSCs) and remove such an inhibitory effect by the small interfering RNA (siRNA) mediated knockdown of the Nogo66 receptor (NgR). Methods NSCs derived from the rat spinal cord were collected, and were cultured by the suspension culture in vitro. NSCs were transfected by siRNA to knock downtheexpression of NgR. Immunofluorescence and Western blot were used to assess the knockdown efficiency. NSCs were divided into four groups and differentiated in the medium containing 10% FBS. In the control group, no intervention was applied to NSCs; in the Nogo-P4 group, NSCs were differentiated in the presence of Nogo-P4 (active segment of Nogo-66); in the siRNA group, NSCs were transfected by siRNA to knock down NgR before they were differentiated; in the siRNA and Nogo-P4 group, NSCs were transfected by siRNA to knock down NgR before they were differentiated in presence of Nogo-P4. The differentiated neurons were labeled by immunofluorescence, and the neurite length was measured by the ImagePro Plus 5.0 software. The differentiation of the neurite length was compared in each group. Results The suspension-cultured cells became the nerve bulb, which could positively expresses Nestin by immunofluorescence. At 1 week of the differentiation in the medium containing 10% FBS, the positively-labeled neuron specific enolase, the glial fibrillary acidic protein, and the myelin basic protein were observed. Both immunofluorescence and Western blot approved that the expression of NgR was knocked down by transfection of siRNA at 24 hours after the transfection. The knockdown efficiency was 90.35%±3.10%. The neurite length was 97.80±6.97 μm, 80.54±6.75 μm,92.14±7.27 μm, and 94.01±8.37 μm in the control group, the Nogo-P4 group, the siRNA group, and the siRNA and Nogo-P4 group, respectively. The Nogo-P4 group had a significant difference when compared with the otherthree groups (Plt;0.01), and the other three groups had no significant difference when compared with each other（Pgt;0.05）. ConclusionNogo-66 can inhibit the neuronal neurite outgrowth during the differentiation ofNSCs. Such an inhibitory effect can be removed by the siRNA mediated knockdown of NgR.
Objective To discuss the stabil ity and practical ity of temporomandibular joint replacement by establ ishing goats artificial temporomandibular joint replacement model. Methods Six healthy mature goats were selected, the male and female being half and weighing 35.3-37.0 kg. According to the parameters from X-ray films of goat’ s temporomandibular joint and the shape of the same kind goat’s skull, the total temporomandibular joint prosthesis was prepared. The one side temporomandibular joints of six goats were replaced by prosthesis randomly as the experimental group (n=6, fossa and condyle according to replacement location) and the other side by titanium plate as the control group (n=6). At 4,8, and 12 weeks, the histological observation, scanning electron microscope (SEM) observation were carried out for observing structural changes in the interface. The mechanical test and histochemistry test were used for observing the combination degree of interface and the alkal ine phosphatase (ALP) activity. Results All animals were al ive to the end of experiment with normal open mouth, good recovery of masticatory function, and normal eating. At 4, 8, and 12 weeks, implants were stable in 2 groups without loosening. The histological observation and SEM observation showed the amount of osteoblasts in interface increased over times. There were significant differences in the shearing force and the ALP activity between fossa in experimental group and control group at 4 weeks (P lt; 0.05), but there was no significant difference between other groups (P gt; 0.05). Conclusion The total temporomandibular prosthesis has good stabil ity in temporomandibular joint reconstruction of goat after replacement.
Objective To observe the effects of immunologic cytokines or anti-angiogenesis gene transfer mediated by electroporation for choroidal melanoma cells.Methods The human embryo kidney cells and malignant choroidal melanoma cells were transfected with plasmids pNGVL-mIL2, pNGVL-mIL12, pCI-sFLK-1, pCR3.1-antiVEGF121,pCI-ExTek. Then the expression of mIL2, mIL12, sFLK-1, VEGF and ExTek were detected by enzymelinked immunosorbentassay (ELISA) and Western blot. Nude mice models of malignant choroidal melanoma were established and they were divided into four groups randomly. Each group was treated with 30 mu;l of 0.9% NaCl, 30 mu;g pNGVL, 30 mu;g antiVEGF121+sFLK-1+ExTek and 30 mu;g mIL2+mIL12 respectively by electroporation. Seven，14, 21, 28, 35 and 42 days after treatment, the tumor volumes were measured to calculate the tumor inhibition rate. Results ELISA and Western blot showed that mIL2，mIL12，sFLK-1 and ExTek were expressed after electroporation，VEGF expression was decreased remarkably. After treatment，the tumors of mIL2+mIL12 group were greatly inhibited with a tumor inhibition rate of 97.33%，while the tumors of antiVEGF121+sFLK-1+ExTek and pNGVL group were partially inhibited with tumor inhibition rates of 53.33% and 36.33% respectively.Conclusions Immunologic cytokines transfer mediated by electroporation can inhibit the growth of melanoma，but anti-angiogenesis only have a mild effects.
Objective To explore the relationship between imbalance in sagittal plane as well as structural factors and lumbar degenerative disease. Methods Patients diagnosed between July 2012 and May 2015 were divided into 4 groups according to corresponding diagnostic criteria: lumbar disc herniation group (LDH), lumbar disc protrusion group (LDP), degenerative lumbar spondylisthesis group (DLS) and nonspecific low back pain group (NLBP); 40 patients were included in each group according to their visiting time. All patients underwent X-ray, CT, and MRI. Sagittal parameters and evaluate degeneration level of structural factors were measured, and the difference among the groups were analyzed. Results There was statistical significance in differences of pelvic incidence (PI) and lumbar lordosis (LL) among 4 groups (P<0.05). Average PI was followed in descending order: DLS, LDP, NLBP, and LDH; average LL was followed in descending order: DLS, NLBP, LDP, and LDH. There was no statistical differences in sacral slope and pelvic tilting among 4 groups (P>0.05). The difference in the level of lumbar disc degeneration between NLBP group (which had slightest lumbar disc degeneration) and the other groups was significant (P<0.001) while no statistical differences in level and rate of lumbar disc degeneration among the other three groups was found (P>0.05). As to the level of lumbar zygapophyseal joint degeneration, there was statistical differences between NLBP group (which had the lowest level of lumbar zygapophyseal joint degeneration) and the other groups (P<0.001) while no statistical differences in the grade of lumbar zygapophyseal joint degeneration among the other three groups (P>0.05). There was statistical differences in the rate of lumbar zygapophyseal joint degeneration between LDH and DLS group (χ2=11.429,P=0.001). Conclusions Vertical lunbar spine is combined with LDH of which the level of lumbar zygapophyseal joint degeneration is minimized, while crooked lunbar spine is combined with DLS of which the level of lumbar zygapophyseal joint degeneration is maximization. NLBP has the lowest level of degeneration of lumbar disc and lumbar zygapophyseal joint degeneration.
ObjectiveTo explore the advantages and disadvantages of using two intracranial EEG (iEEG) monitoring methods—Subdural ectrodes electroencephalography (SDEG)and Stereoelectroencephalography (SEEG), in patients with “difficult to locate” Intractable Epilepsy. MethodsRetrospectively analyzed the data of 60 patients with SDEG monitoring (49 cases) and SEEG monitoring (11 cases) from January 2010 to December 2018 in the Department of Neurosurgery of the First Affiliated Hospital of Fujian Medical. Observe and statistically compare the differences in the evaluation results of epileptic zones, surgical efficacy and related complications of the two groups of patients, and review the relevant literature. ResultsThe results showed that the two groups of SDEG and SEEG had no significant difference in the positive rate and surgical resection rate of epileptogenic zones, but the bilateral implantation rate of SEEG (5/11, 45.5%) was higher than that of SDEG (18/49, 36.7%). At present, there was no significant difference in the postoperative outcome among patients with epileptic zones resected after SDEG and SEEG monitoring (P>0.05). However, due to the limitation of the number of SEEG cases, it is not yet possible to conclude that the two effects were the same. There was a statistically significant difference in the total incidence of serious complications of bleeding or infection between the two groups (SDEG 20 cases vs. SEEG 1 case, P<0.05). There was a statistically significant difference in the total incidence of significant headache or cerebral edema between the two groups (SDEG 26 cases vs. SEEG 2 cases, P<0.05). There was a statistically significant difference in the incidence of cerebrospinal fluid leakage, subcutaneous fluid incision, and poor healing of incision after epileptic resection (SDEG 14 cases vs. SEEG 0 case, P<0.05); there were no significant differences in dysfunction of speech, muscle strength between the two groups (P>0.05). ConclusionSEEG has fewer complications than SDEG, SEEG is safer than SDEG. The two kinds of iEEG monitoring methods have advantages in the localization of epileptogenic zones and the differentiation of functional areas. The effective combination of the two methods in the future may be more conducive to the location of epileptic zones and functional areas.