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.

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.

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, P<0.05), with the more area of spinal cord remained (F>49.990, P<0.05). There were less Oct3/4 positive cells in the CE3-Mash-1 group than CE3 group (t=5.439, P<0.001), with more nestin (t=-7.536, P<0.001) andβ-tubulin III (t=-9.941, P<0.001) positive cells. However, there was no significant difference in GFAP positive cells between CE3-Mash-1 and CE3 groups (t=1.701, P>0.05). Conclusion Overexpression of Mash-1 gene promotes CE3 cells to differentiate into neurons in spinal cord injury mice, and improve the motor function recovery.

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.

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.

Purpose:To study the relationship between the expression of CD44v6 with the clinicopathological charac- teristics and the prognosis in primary gastric carcinoma patients.Methods:A total of 188 paraffin-embedded gastric carcino- mas and 42 non-carcinomatous gastric mucosae was stained with the monoclonal antibodies CD44v6 using the EnVision~(TM) method.Results:The expression level of CD44v6 were significantly higher in the tumors (67.6%) than in the non-carcino- matous gastric mucosae (9.5%) (P

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.

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.

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 (ratio<0.5) as compared with the normal control group. On the other hand, in the Yiqi Huayu Recipe group, 97 kinds of genes expressed in cervical intervertebral disc chondrocytes were screened out, in which 44 were up-regulated (ratio>2) 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.

This study was aimed to optimize bio-solid bidirectional fermentation conditions of Trametes robiniophia Murr. for rhubarb. Sulphuric acid-phenol colorimetry, magnesium-methanol colorimetry, and HPLC were used in the
content determination of polysaccharide, total anthraquinone, and 4 free anthraquinones. The drying rate and con-sumption rate were combined as indicators for the optimization of technical parameters such as medicinal dosage, temperature and amount of water. The results showed that when using 500 mL conical flask, the best fermentation conditions were medicinal dosage of 10 g, the temperature of 34℃, adding water of 120%. It was concluded that bidirectional fermentation of rhubarb increased the content of free anthraquinones. Among them, the content of chrysophanol with anti-oxidation effect increased significantly. The decreasing of combined anthraquinone can relieve the severe laxative effect of rhubarb.