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This study was aimed to investigate the effect of traditional Chinese medicine (TCM) compound "Ji-Sui-Kang (JSK)" on the differentiation of neural stem cells (NSCs).Drug serum of JSK was prepared.The primary NSCs were isolated and cultured.NSCs were identified.Fetal bovine serum (FBS) was used to induce differentiation of NSCs.Different doses of drug serum of JSK was added to intervene the differentiation.There were the high-,medium-,and low-dose group.The blank control group was also set.The β Ⅲ-tubulin and GFAP were used to detect the differentiation of NSCs.And then,it was compared with the control group without drug serum.Positive cells ofβ m-tubulin and GFAP in different visual fields were counted.And the percentage of positive cells in the total number of cells was calculated.Fluorescence microscope was used to conduct quantitative fluorescence experiments.The results showed that the cultured neurospheres were expressed of Nestin protein.The percentage ofβⅢ-tubulin positive cells in the high-,medium-and low-dose group of drug serum of JSK was significantly higher than that in the control group (P＜0.05).The percentage of GFAP positive cells in the high-,medium-and low-dose groups of drug serum of JSK was significantly lower than that in the control group (P＜0.05).And the high-dose group of drug serum of JSK was statistically significant compared with the middle-and low-dose group of drug serum of JSK (P＜0.01).It was concluded that TCM compound JSK promoted thedifferentiation of NSCs intoβⅢ-tubulin positive cells.It provided a basis for treatment of SCI with TCM.
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Objective To investigate the effect of Chinese herbal compound"Jisuikang"on the phagocyto-sis of neuronal debris by microglial cells. Methods To prepare serum containing drugs of JSK and divide them into the low,middle and high dose groups,the blank serum group and LPS+blank serum group. BV2 was labeled by lentiviral vectors containing the green fluorescent protein gene (GFP). To establish the damage neuron model and mix injured neurons with the transfected microglia. To observe the situation of microglia which was affected by serum containing drugs devour the neuronal debris. Results The middle and high dose of JSK showed greater phagocytic percentage and phagocytic index than those of the control group(P<0.001). In comparison of LPS+blank serum group,no significant difference was found in the middle and high dose of JSK. However,to the phagocytic index, which was better than that of LPS+blank serum group(P<0.05). Conclusion JSK may enhance the engulfment of neuron debris by BV2,which could provide a better living environment for the growth of neurons.
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Objective To investigate the effect of Chinese herbal compound"Jisuikang"on the phagocyto-sis of neuronal debris by microglial cells. Methods To prepare serum containing drugs of JSK and divide them into the low,middle and high dose groups,the blank serum group and LPS+blank serum group. BV2 was labeled by lentiviral vectors containing the green fluorescent protein gene (GFP). To establish the damage neuron model and mix injured neurons with the transfected microglia. To observe the situation of microglia which was affected by serum containing drugs devour the neuronal debris. Results The middle and high dose of JSK showed greater phagocytic percentage and phagocytic index than those of the control group(P<0.001). In comparison of LPS+blank serum group,no significant difference was found in the middle and high dose of JSK. However,to the phagocytic index, which was better than that of LPS+blank serum group(P<0.05). Conclusion JSK may enhance the engulfment of neuron debris by BV2,which could provide a better living environment for the growth of neurons.
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BACKGROUND: Myelin sheath related inhibitors have been found to have great impact on microenvironment of axon regeneration. Traditional Chinese medicine is gradual y becoming a research hotspot on improving microenvironment of nerve regeneration with its advantage on multiple factors and targets. OBJECTIVE: To clarify the effect of Jisuikang on Nogo-66 receptor NgR expression after spinal cord injury. METHODS: 144 rats were randomly divided into six groups: sham surgery group, model group, prednisone group, high-, moderate- and low-dose Jisuikang groups (n=24). Animal models of spinal cord injury were established by the modified Allen’s method in the later five groups. Rats in the prednisone group were daily given 0.06 g/kg prednisone acetate by lavage, once a day. Rats in the high-, moderate- and low-dose Jisuikang groups were daily intragastrically given 12.5, 25 and 50 g/kg Jisuikang, once a day. Rats in the sham surgery and model groups were intragastrically daily given 20 mL of saline, once a day. Rats in each group were administered drugs until death. RESULTS AND CONCLUSION: Compared with the model group, NgR protein and mRNA expression levels were significantly lower in the prednisone, moderate-and low-dose Jisuikang groups. These data suggested that Jisuikang can improve the recovery of neurological function after spinal cord injury and effectively inhibit NgR protein expression at the site of injury so as to suppress the microenvironment factors harmful to nerve regeneration and further improve the microenvironment of nerve regeneration. Subject headings: Drugs, Chinese Herbal; Spinal Cord Injuries; Axons; Tissue Engineering
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Objective To clarify the effects of Jisuikang on expression levels of BDNF and NGF protein in serum of rats with spinal cord injury;To explore its probable mechanism to improve the axon regeneration microenvironment and resistance spinal cord injury.Methods Ninety SD rats were chosen to establish acute spinal cord injury model through modified Allen's method. 15 rats were chosen randomly as sham-operation group, and the other 75 rats were randomly divided into model group, prednisone group and high-, medium-, and low-doseJisuikang groups. All administration groups were given relevant medicine for gavage, while rats in sham-operation group and model group were given normal saline with the same volume for gavage. The activity and death condition of rats were recorded. Rats were put to death on the 3rd, 7th and 14th d of intervention for carotid artery blood collection. The expressions of BDNF and NGF in serum were detected by ELISA.Results After modeling, the rats in model group showed a typical paraplegia syndrome. 3 day after operation, BDNF showed no statistical significance between model group and prednisone group, while each treatment group showed significant difference when compared with prednisone group (P<0.05,P<0.01). 7 d and 14 d after operation, compared with the model group, BDNF of prednisone group and medium-dose Jisuikang group showed statistical significance, and there was no significant difference between prednisone group and medium-dose Jisuikang group (P<0.05,P<0.01). The expression of NGF in each treatment group at each time point with the prednisone group showed statistical significance. 3 d and 7 d after operation, among prednisone group, high- and medium-doseJisuikang groups, the expression of NGF was significantly higher than model group. 14 d after operation, compared with the model group, the expression of NGF in medium- and low-dose Jisuikang groups and prednisone group still maintained at a relatively high level, with statistical significance (P<0.05,P<0.01).Conclusion Jisuikang can effectively promote the expressions of BDNF and NGF in serum after spinal cord injury and maintained at a relatively high level, so as to improve the microenvironment of axonal regeneration and promote nerve regeneration.
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Objective To explore the effect of Jisuikang on neural functional recovery, and expression of brain-derived neurotrophic fac-tor (BDNF) protein and mRNA level after spinal cord injury (SCI). Methods 144 female Sprague-Dawley rats, weighted 180 to 220 g, were used for experiment. 24 rats were randomly extracted into sham group (Group A), which had their vertebral plates and spines bitten away on-ly. The others were randomly divided into model group (Group B), prednison group (Group C), and high, middle and low doses of Jisuikang group (Groups D to F) after SCI, 24 rats in each group. Group C was given 0.06 g/(kg?d) prednison, and Groups D to F were given 50, 25 and 12.5 g/(kg?d) Jisuikang respectively, which were given 20 ml/(kg?d) volume by intragastric administration. Groups A and B were given the same volume of normal saline (NS). The Basso-Beattie-Bresnahan (BBB) scores and oblique board test were applied to test the postoper-ative results 24 hours, 3, 7 and 14 days after SCI. The rats were executed and the spinal cord tissues were extracted 3, 7 and 14 days after SCI. Immunohistochemistry, Western blotting and RQ-PCR were applied to test the expression of protein and mRNA of BDNF. Results BBB scores and angle of oblique board test were significantly lower in Groups B to F than in Group A 24 hours after SCI (P0.05). The results of RQ-PCR showed that prednisone and Jisuikang promot-ed the expression of BDNF mRNA. Group C (prednisone) had a most obvious effect at the beginning while Group E was better than Group C 14 days after SCI. Conclusion Jisuikang can promote the neural functional recovery and the expression of BDNF on both protein and mRNA level in SCI rats.
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BACKGROUND:Studies have shown that Clematis chinensis Osbeck can promote articular chondrocyte proliferation, maintain and promote the synthesis of cartilage proteoglycan and type II col agen in chondrocytes. Low-intensity pulsed ultrasound can effectively improve the carticular chondrocyte proliferation, increase the cellmembrane permeability, and promote chemicals or genes delivery, so as to improve the biological effects of chemicals. OBJECTIVE:To observe the effect of low-intensity pulsed ultrasound mediated Clematis chinensis Osbeck on the proliferation of cartilage cells and the expression of type II col agen and transforming growth factor (TGF)-β1 of rabbit knee articular chondrocytes cultured in vitro, and explore the effect and mechanism of Clematis chinensis Osbeck mediated by low-intensity pulsed ultrasound on cartilage damage. METHODThe rabbit knee articular chondrocytes were isolated and cultured in vitro. cells of the second generation in logarithmic growth phase were randomly divided into four groupcontrol group, low-intensity pulsed ultrasound (LIPUS) group, Clematis chinensis Osbeck (CCO) group, and CCO+LIPUS group. After cells were cultured under different conditions for 3 days, cellproliferation was measured with cellCount Kit-8 assay. The secretion of type II col agen was detected by cytochemical staining method, and the expression of TGF-β1 was assayed by western blot analysis. RESULTS AND CONCLUSION:The number of cells in the other three groups were significantly higher than that in control group at 3 days after culture (P<0.05), while the CCO+LIPUS group had the most cells. Cytochemical staining showed that the area and absorbance value of type II col agen in cartilage cells were significantly increased in CCO+LIPUS group, compared with control group (P<0.05), and the difference with control group was larger than the amount of that of LIPUS group and CCO group with control group. Western blot analysis showed that TGF-β1 was significantly expressed in CCO+LIPUS group, and the relative gray value was significantly higher than the other groups (P<0.05). Both low-intensity pulsed ultrasound and Clematis chinensis Osbeck can promote the proliferation of cartilage cells and expression of type II col agen and t-β1 of rabbit knee articular chondrocytes cultured in vitro. The combined method could produce synergistic effects.
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<p><b>OBJECTIVE</b>To develop an intelligent adjustable leg raising system which could realize the quantification and visualization of lower limbs' elevation angle and frequency.</p><p><b>METHODS</b>We determined the pole adjustable length of thighs and cruses and the flexion angle range of hips and knees according to the requirements of clinical lower limb function rehabilitation, and made force analysis of hip's and knee's flexion motivation mechanism and clinical observation on its security and effectiveness.</p><p><b>RESULTS</b>This device was small and compact in overall appearance, which could adjust the angle of the hips and knees bending and ankles turning alone. The force analysis of the hips and knees flexion power element was consistent with the design requirements. The preliminary studies showed the device could relieve pain, improve the range of motion and promote the rehabilitation, which was superior to that of the Brown Ska (P < 0.05).</p><p><b>CONCLUSION</b>The intelligent adjustable leg raising system meets the requirements of clinical usage, which is suitable for different heights, and the flexion angle ranges of hips and knees are wide and in high accuracy, which is worth of being improved and generalized.</p>
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Humans , Leg , Manipulation, Orthopedic , Range of Motion, Articular , RehabilitationABSTRACT
There exist some problems such as faultiness in system,illogicality in constructing of team,the simple ways and means in teaching inspection.Therefore,the universities in our country must take such measures as system healthiness,team construction,and mechanism function improvement.