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Objective To investigate the effects of overexpression of Mash-1 gene on functional recovery and neural differentiation of embryonic stem cells in spinal cord injury mice. Methods CE3 cell line with overexpression of Mash-1 gene was generated with murine stem cell virus. Spinal cord injury model was established with forceps compression in 4-week-old KM mice. Normal saline (model group, n=12), CE3 cells with or without overexpression Mash-1 gene (CE3-Mash-1 and CE3 groups, n=12 respectively) were transplanted into the ar-eas of injury 3 days after injury. They were assessed with the Basso Mouse Scale (BMS) 1, 7, 14, 21, and 28 days after injury. 6 mice from each group were sacrificed 14 and 28 days after injury respectively. The spinal cord area remained were observed with HE stained, and the expression of Oct3/4, nestin,β-tubulin III and glial fibrillary acidic protein (GFAP) were detected with immunofluorescence in the injured spinal cord in the CE3 and CE3-Mash-1 groups. Results The score of BMS significantly improved in CE3 and CE3-Mash-1 groups com-pared with that of the model group (F>84.471, P49.990, P0.05). Conclusion Overexpression of Mash-1 gene promotes CE3 cells to differentiate into neurons in spinal cord injury mice, and improve the motor function recovery.
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10.3969/j.issn.2095-4344.2013.24.016
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Professor Shi Qi is a famous traditional Chinese medicine doctor specializing in orthopaedics and traumatology, who has formatted a set of systematic protocols for the diagnosis and treatment of chronic musculoskeletal conditions. When it is time for using tonics in winter, he advocates applying herbal paste for treating chronic musculoskeletal diseases. This paper introduces Professor Shi Qi's commonly used prescription for treating chronic musculoskeletal conditions and puts forward demands and understandings in concocting herbal paste, experience in herbal paste for treating chronic musculoskeletal diseases such as cervical spondylosis, lumbar disc herniation, lumbar spinal stenosis, lumbar muscle strain, ankylosing spondylitis, osteoporosis, knee osteoarthritis and avascular necrosis of femoral head, and the advantages of herbal paste for treating chronic musculoskeletal conditions as opposed to alternative treatments.
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Osteoarthritis is one of the most common diseases seen in clinical practice. Cartilage survives in the hypoxic microenvironment. Hypoxia-inducible factor-1α (HIF-1α) is a key nuclear transcription factor which mediates the hypoxic response of cells. HIF-1α gene is an important regulator for the adaptation of articular cartilage to the hypoxic environment. It is important for formation of articular cartilage, energy metabolism and matrix synthesis. If the HIF-1α gene is knocked out, the cartilage can not maintain their normal morphology and function, which may lead to cartilage degeneration, and result in diseases such as osteoarthritis. Chinese herbal medicines can regulate the expression of HIF-1α gene and supply a therapy method for osteoarthritis. In this paper, the authors review the situation of the correlation between HIF-1α and osteoarthritis cartilage degeneration examined in recent years.
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To observe the effects of Yiqi Huayu Recipe, a Chinese compound herbal medicine, on apoptosis of dorsal root ganglion (DRG) neurons and expression of caspase-3 in rats after lumbar nerve root compression injury.
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To observe the effects of Yiqi Huayu Decoction, a compound traditional Chinese herbal medicine, on brain-derived neurotrophic factor (BDNF) expression in lumbar nerve root and soleus of rats after lumbar nerve root compression injury.
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To explore the mechanism of Yiqi Huayu Bushen Recipe (YHBR), a compound traditional Chinese herbal medicine, in treating cervical syndrome (CS) with qi deficiency, blood stasis and kidney deficiency in rats.
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To research the action mechanism of Yiqi Huayu Bushen Recipe (YHBR), a compound of traditional Chinese herbal medicine, in treating cervical syndrome (CS) with kidney deficiency in rats.
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To establish a rat model of cervical syndrome with qi deficiency, blood stasis and kidney deficiency.
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OBJECTIVE: To establish a rat model of cervical syndrome (CS) with kidney deficiency. METHODS: A group of 30 three-month-old female Sprague-Dawley rats were randomly divided into normal control group, CS group and CS with kidney deficiency group (combined group), with 10 rats in each group. Rats in the normal control group received no treatment, rats in the CS group underwent only resection of cervical muscles and ligaments as unbalanced dynamic and static animal model, and rats in combined group underwent resection of both cervical muscles and ovaries as kidney deficiency model. Serum and cervical intervertebral discs were collected. Kidney deficiency was determined by observing the morphologic changes of uterus and appendages, detecting the weight of uterus and appendages and the content of serum estradiol (E(2)). The degeneration of intervertebral discs was determined by detecting the histopathology, the expressions of type II collagen and type X collagen proteins, and the expressions of aggrecan-1 (Agc1), type II procollagen gene (Col2a1), matrix metalloproteinase-13 (MMP-13) and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNAs. RESULTS: Compared with those in the normal control group and CS group, the rats in the combined group were noted with the uterus atrophied, the caliber of oviduct thinned, the weight of uterus and appendages diminished obviously, the content of serum E(2) decreased, cervical intervertebral disc degenerated more seriously, type II collagen protein expression decreased, type X collagen protein expression increased, Agc1 and Col2a1 mRNA expressions in intervertebral disc decreased, and the MMP-13 mRNA expression increased. CONCLUSION: The rat model of CS with kidney deficiency is established. Kidney deficiency can aggravate cervical intervertebral disc degeneration.
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OBJECTIVE: To make an animal model of cervical spondylosis (arthralgia syndrome type) with stimulation of wind, cold, and dampness. METHODS: Twenty-four 8 months old male New Zealand white rabbits were randomly allocated into four groups: normal control group, light stimulation group, moderate stimulation group and severe stimulation group. The wind speed was 10.8-13.8 m/s, the temperature was (5+/-0.5)degrees centigrade, and the humidity was 100%. The rabbits of light, moderate, and severe stimulation groups were kept in the above-mentioned environments for 4 hours everyday, and for a total of 32, 64, and 128 hours, respectively. The intervertebral discs were stained with HE method, and observed with a light microscope. Prostaglandin E(2) (PGE(2)), 6-ketone-prostaglandin F1alpha (6-K-PGF(1alpha)) and thromboxane B(2) (TXB(2)) contents were measured by ELISA. Fas and Bcl-2 expressions were examined by immunohistochemical avidin-biotin peroxidose complex technique. Interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), and transforming growth factor beta (TGF-beta) mRNA expressions were examined by reverse transcription-polymerase chain reaction. RESULTS: The nucleus pulposus of rabbits in the light and moderate stimulation groups shrunken, and in the severe stimulation group, the anulus fibrosus loosed or ruptured, and the cartilage end-plate became proliferated. Compared with rabbits in the normal control group, the PGE(2) content rose in the light stimulation group, the contents of PGE(2), 6-K-PGF(1alpha), and TXB(2) increased, the expressions of IL-1beta and TNF-alpha mRNAs and Fas were up-regulated, and the expressions of TGF-beta mRNA and Bcl-2 were down-regulated in the moderate and severe stimulation groups. The expression of Fas was up-regulated mostly and Bcl-2 was down-regulated mostly in the severe group. CONCLUSION: Moderate and severe stimulations of wind, cold and dampness can lead to degeneration of cervical intervertebral discs of rabbits. The model corresponds to the theory of traditional Chinese medicine about arthralgia syndrome caused by wind, cold and dampness.
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OBJECTIVE: To set up a natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats in order to offer an appropriate carrier for the study on mechanism of intervertebral disc degeneration. METHODS: The method of enzyme digestion combined with natural subculture was used to set up the in vitro natural degeneration model of chondrocytes derived from the endplate of intervertebral disc of rats. The morphological appearances and microstructures of the chondrocytes of different generations were observed. The expression of collagen II in chondrocytes was detected by immunocytochemical method. RESULTS: The chondrocytes derived from the endplate of intervertebral disc expressed collagen II. After 13 days of culture, the chondrocytes of generation III showed that the ability of cell division descended, the nucleoli became unclear, the cells deformed obviously, fusiform shape with weak optical activity appeared, and the intercellular space was enlarged. There were vacuoles and lipid droplets in cytoplasm. The synthesis of collagen II, as well as the cell proliferation rate, descended notably. All results showed the natural degeneration process of the chondrocytes. CONCLUSION: The in vitro natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats was successfully established. This can offer the cytological basis for study on the mechanism of intervertebral disc degeneration.
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OBJECTIVE: To study the effects of Yiqi Huayu Recipe on neural cell adhesion molecule (N-CAM) in neuromuscular junctions during nerve regeneration in rats with lumbar nerve root compression. METHODS: The rats with lumbar nerve root compression were given Yiqi Huayu Recipe for 10, 20 and 30 days respectively. The distribution of N-CAM in neuromuscular junctions of soleus muscle in rats was examined with immunohistochemical method and confocal laser scanning microscopy technique. The acetylcholine receptor (AChR) was visualized with fluorescein-conjugated alpha-bungarotoxin (alpha-BTX). The overlap areas of N-CAM and AChR sites were measured with NIH image technique. RESULTS: The aggregates, sprouts and extensions of N-CAM in the neuromuscular junctions and the overlap areas of N-CAM and AChR sites in the Yiqi Huayu Recipe-treated group were all better improved than those in the untreated group. CONCLUSION: The expression of N-CAM is regulated according to the state of innervation for muscles. Yiqi Huayu Recipe may accelerate this nerve regeneration process.
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To investigate the gene expression changes in the degenerated cervical intervertebral discs of rats, and to study the function of Yiqi Huayu Bushen Recipe, a compound Chinese herbal medicine, and its discomposed formulas in regulating gene expressions in the degenerated cervical intervertebral discs.
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OBJECTIVE: To study the effects of Yiqi Huayu Recipe on the gene expression pattern in normal and apoptotic chondrocytes in the cervical intervertebral disc of rats. METHODS: The intervertebral disc endplates of rats were digested enzymatically. The apoptosis of the chondrocytes was induced by anti-Fas antibody. BiostarR-40s microarray chips were used to investigate the gene expression pattern in chondrocytes in the normal control, apoptosis and Yiqi Huayu Recipe groups. The results were scanned with Scan Array 4000 and analyzed with GenePix Pro 3.0, and then subjected to standardization and ratio analysis. RESULTS: In the apoptosis group, 30 kinds of genes expressed in cervical intervertebral disc chondrocytes were screened out, in which 10 were up-regulated (ratio>2) and 20 down-regulated (ratio2) and 53 down-regulated (ratio<0.5) as compared with the apoptosis group. CONCLUSION: There are some signal transduction pathways that control the apoptosis of the intervertebral disc chondrocytes. Yiqi Huayu Recipe regulates the gene express of the apoptotic chondrocytes in intervertebral disc. The study gives a further understanding to the mechanism of the cervical spondylosis and enriches the theories of qi and blood of traditional Chinese medicine.
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OBJECTIVE: To study the effects of Yiqi Huayu Recipe and its decomposed formulas-medicated sera on expressions of bcl-2, Bax and caspase-8 of apoptotic anulus fibrosus cells in rats. METHODS: Immunohistochemical and integral optical density analytic methods were used to observe the effects of Yiqi Huayu Recipe-, Yiqi Recipe-, Huayu Recipe-medicated sera and insulin-like growth factor 1 (IGF-1) on the expressions of bcl-2, Bax and caspase-8 of apoptotic anulus fibrosus cells in rats induced by anti-Fas antibody. RESULTS: As compared with apoptosis group, bcl-2 expression was higher, Bax and caspase-8 expressions were lower in Yiqi Huayu-treated, Yiqi-treated, Huayu-treated and IGF-1 groups (P<0.01). As compared with Yiqi-treated group and Huayu-treated group, Bax expression was lower in Yiqi Huayu-treated group (P<0.05). CONCLUSION: Yiqi Huayu Recipe and its decomposed formulas can delay degeneration of the cervical intervertebral disc, which may be due to its action in regulating the expressions of bcl-2, Bax and caspase-8.
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OBJECTIVE: To study the effect of Yiqi Huayu Recipe on peanut agglutinin-binding molecules (PNA-BMs) and Schwann's cells (Sc) of the neuromuscular junction after L5 nerve root compression in rats. METHODS: The rats were given Yiqi Huayu Recipe at the 10th, 20th, 30th and 60th day after L5 nerve root compression. Using immunohistochemistry and confocal laser scanning techniques, we observed the distributions of PNA-BMs and S-100 in the soleus muscle. The overlap area of PNA-BMs and S-100 was measured with NIH image technique. RESULTS: The extracellular matrix (ECM) recognized by PNA played an important role in guiding the sprouting and extending of the Schwann's cells. The agglomeration, sprouting and extension of the terminal Sc and the overlap area of Sc with PNA-BMs in the Yiqi Huayu Recipe group were better than those in the control group. CONCLUSION: Yiqi Huayu Recipe can promote the growth of ECM and Schwann's cells and accelerate the nerve regeneration process.
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Ligusticum chuanxiong is a Common Chinese medicinal herb which is widely used in clinical treatment. Modern research has shown that the ingredients of Ligusticum chuanxiong such as tetramethylpyrazine and ferulic acid have multiple effects of scavenging oxygen free radicals,calcium-antagonism,vasodilating,inhibiting platelet aggregation and thrombosis and so on. This article reviews the research progress in pharmacological activities of the available compositions in ligusticum chuanxiong in the past several years.
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OBJECTIVE: To study the effect of Yiqi Huayu Recipe on neurons in neuromuscular junction after L5 nerve root compression in rats. METHODS: By using immunohistochemistry, confocal laser scanning techniques and protein gene product 9.5 as a neuronal marker, the motor endplates were visualized with fluorescein-conjugated alpha-BTX. The effect of Yiqi Huayu Recipe on nerve terminals of neuromuscular junction at 10, 20, 30, 60 days after L5 nerve root compression was observed in rat soleus muscles during regeneration. The overlap areas of terminals and endplates were measured with NIH image technique. RESULTS: As for the aggregates, sprouts, extension of nerve terminal during regeneration, the connecting speed and range of nerve terminal with endplates, as well as the reformation of neuromuscular junctions during regeneration, Yiqi Huayu Recipe group was better than that of the control group. CONCLUSION: Yiqi Huayu Recipe can promote the hyperplasia of neurons and enhance the regeneration function. It also can accelerate the neuromuscular junction reformation and shorten the process of regeneration.
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OBJECTIVE: To study the effects of Gegen Recipe and Guizhi Recipe regulating the expression of IL-1beta, iNOS, TNFalpha and TGFbeta mRNA in degenerated intervertebral disk caused by stimulation of wind, cold and damp. METHODS: Thirty-six rabbits were randomly separated into six groups, viz. control group, stimulation by wind, cold and damp group, head in low-location group, compound model group (head in low-location plus stimulation of wind, cold and damp), Gegen Recipe group and Guizhi Recipe group. The gene expression of IL-1beta, iNOS, TNFalpha and TGFbeta mRNA in degenerated cervical intervertebral disks were assayed by RT-PCR. RESULTS: Compared with the control group, IL-1beta mRNA and iNOS mRNA expression were up-regulated in each model group(P<0.01). Both Gegen Recipe and Guizhi Recipe down-regulated IL-1beta mRNA, iNOS mRNA expression, which had significant differences compared with the stimulation model group(P<0.01 or <0.05). TNFalpha mRNA expression was significantly enhanced in the compound model group compared with the control group (P<0.01). There was no significant difference between the head in low-location group and the stimulation model group. Gegen Recipe down-regulated TNFalpha mRNA expression compared with the stimulation group(P<0.01),while the effect of Guizhi Recipe was not significant. Compared with the control group, TGFbeta mRNA expression went down in each model group(P<0.01). Gegen Recipe up-regulated TGFbeta mRNA expression(P<0.01), while the effect of Guizhi Recipe on adjusting TGFbeta mRNA was not significant. CONCLUSION: Stimulation by wind, cold and damp, head in low-location, and compound of both can down-regulate the expressions of IL-1beta, iNOS, TNFalpha mRNA, and up-regulate the expression of TGFbeta mRNA. These researches certify that these factors take part in the progression of intervertebral disk degeneration. IL-1beta, iNOS, TNFalpha accelerate the degeneration and TGFbeta defers the degeneration. Gegen Recipe down-regulates the expression of TNFalpha mRNA and up-regulates the expression of TGFbeta mRNA. The effect on regulating the cell factors expressions by Guizhi Recipe is lower than that of Gegen Recipe.