Establishment of a chondrocyte degeneration model by over-expression of human Wnt7b gene in 293ft cell line / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of human Wnt7b gene in rat chondrocyte degeneration.</p><p><b>METHODS</b>Wnt7b gene obtained by PCR was cloned to PCDH-GFP. 293ft cell line was transfected with PCDH-GFP and PCDH-Wnt7b, and the supernatant and transfected cells were collected. The expression level of Wnt7b in 293ft cells was detected by Western blotting. The first passage of chondrocytes were isolated from articular cartilages of newborn born (within 24 h) SD rats were cultured in the supernatants from the transfected cells (at 10- and 50-fold dilutions). The cell morphology of the rat chondrocytes was observed under inverted microscope, and the protein expressions of MMP13, MMP3 and type II collagen and mRNA expressions of A-can, ADAMTS5, Col X and Sox9 were examined by Western blotting or real-time PCR.</p><p><b>RESULTS</b>Human Wnt7b gene cloned to PCDH-GFP was expressed efficiently in 293ft cell line. Rat chndrocytes cultured for 24 h in the supernatants from PCDH-Wnt7b-transfected 293ft cells underwent changes from a polygonal to a spindle-shaped morphology. The protein expression levels of MMP13 and MMP3 increased while type II collagen decreased significantly, and the mRNA levels of A-can and Sox9 were down-regulated while Col X and ADMATS5 up-regulated in ratchondrocytes after incubation in supernatants from PCDH-Wnt7b-transfected 293ft cells.</p><p><b>CONCLUSION</b>Human Wnt7b gene can be expressed efficiently in 293ft cell line and can induce rat chondrocyte degeneration in vitro.</p>

Development and validation of a CO-T1 three-dimensional finite element model of a healthy person under physiologic loads / 生物医学工程学杂志

A comprehensive, geometrically accurate, nonlinear C0-T1 three-dimensional finite element (FE) model was developed for the biomechanical study of human cervical spine and related disorders. The model was developed with anatomic detail from the computed tomography (CT) images of a 46-year-old female healthy volunteer, and applied the finite element model processing softwares such as MIMICS13.1, Hypermesh11.0, Abaqus 6.12-1, etc., for developing, preprocessing, calculating and analysing sequentially. The stress concentration region and the range of motion (ROM) of each vertebral level under axial rotation, flexion, extension, and lateral bending under physiologic static loadings were observed and recorded. The model was proven reliable, which was validated with the range of motion in previous published literatures. The model predicted the front and side parts of the foramen magnum and contralateral pedicle and facet was the stress concentration region under physiological loads of the upper spine and the lower spine, respectively. The development of this comprehensive, geometrically accurate, nonlinear cervical spine FE model could provide an ideal platform for theoretical biomechanical study of human cervical spine and related disorders.

Alendronate improves bone matrix structure of ovariectomized rats / 中国组织工程研究

BACKGROUND:Bisphosphonates that can increase bone density and inhibit bone resorption have been clinical y confirmed, but the structure of the bone matrix has been less studied. OBJECTIVE:To observe the effects of alendronate on bone structure and bone matrix metabolism, and then to investigate the control ing mechanism by which alendronate improves bone mass and increase bone intensity. METHODS:An ovariectomized rat model was prepared and intervened with alendronate as treatment group. Model and sham-surgery groups were set as controls. Alendronate effects on bone mineral density, bone metabolism, bone biomechanics, and bone structure were observed in bone loss rats using bone imaging, bone tissue pathology and biomechanical test and enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION:Alendronate intervention could fight against bone loss as compared with model group at weeks 4, 8, and 12 after treatment (P<0.05). Compared with the model group, the expression of urinary deoxypyridinoline and serum carboxyterminal propeptide of type Ⅰ procol agen was decreased significantly after alendronate intervention (P<0.05);the maximal load, maximal pressure and modelus on the lumbar vertebrae and femur were increased as wel as ratio of urinary pyridinoline/deoxypyridinoline of type Ⅰ procol agen (P<0.05). These findings suggest that alendronate intervention can inhibit bone loss in rats induced by estrogen deficiency, increase biomechanical properties, improve bone matrix structure, and meanwhile, recover the Ⅰ col agen crosslinking component due to ovariectomy.

Morphological studies on osteoarthritis of the articular cartilage in C57 black mice and its relationship with etalloproteinase-3 and lnterleukln-1 in serum and synovial fluid / 中华风湿病学杂志

Objective To study the levels of Matrix Metalloproteinase-3 (MMP-3) and interleukin (IL-1) in the synovial fluid and plasma of C57 black mice with osteoarthritis (OA) and their relationships with the severity of pathological changes so as to investigate their effects and correlation with OA. Methods The C57 black mice with OA were enrolled for this study. Different levels of exercise were appicated based on their age. Knee joint pathological changes were examined for pathological severity of OA. ELISA sandwich method was used to measure the levels of MMP-3 and IL-1 in serum and synovial fluid. Correlation analysis was performed to demonstrate the relationship between the levels of MMP-3 and IL-1 in the serum and synovial fluid, and the pathological severity of OA. Results ①Morphological observations: C57 black mice were characterized by spontaneously developing OA and the incidence and the severity of osteoarthritis gradually increased with age and exercising burdens. ② The level of MMP-3 and IL-1 in the synovial fluid of exercising mice MMP-3 (84±6) ng/ml, IL-1 (48±3) ng/ml was higher than that in the aged ones [MMP-3 (84±6) ng/ml, IL-1 (71±5) ng/ml J, the difference was significant (P<0.01). The level of MMP-3 and IL-1 level in the serum had a linear correlation with that of the synovial fluid. At the same time, they also had linear correlation with the pathological severity of OA (All r>0.67, and all P<0.01). Conclusion The levels of MMP-3 and IL-1 in serum and synovial fluid can help to make early diagnosis of OA, especially elevated MMP-3 level.

Comparison of the effect of glucosamine on the cartilage oligomeric matrix protein secretion in vitro chondrocytes and synoviocytes / 中华风湿病学杂志

Objective To compare the effect of glucosamine (Virtral-s) on the cartilage oligomeric matrix protein (COMP) secretion of chondrocytes and synoviocytes in vitro.Methods Chondrocytes and synoviocytes isolated from knee cartilage of osteoarthritic patients were cultured by phased enzymatic digestion.Sera containing Virtral-s of the experimental animals were obtained after orally administrated Virtral-s at the dosages that equal to human.Cells were cultured in the medium with Virtral-s containing sera.Super-natant COMP level was tested by enzyme-linked immunoabsorbent assays (ELISA).Results COMP conceu-tration of synoviocytes cultured in vitro was significantly higher than that of chondrocytes (P＜0.05).Virtral-s could significantly increase COMP secretion in cultured chondrocytes in vitro (P＜0.05),however,it had a weaker role on synoviocytes,ie,it could only mildly reduce COMP secretion of synoviocytes.Conclusion Glucosamine (Virtral-s)-containing serum can promote COMP secretion of chondrocytes in vitro,and it has no significant effect on synoviocytes in vitro.

Effects of Bushen Chinese Medicine on Expression of Cbf?1 in Aging Osteoblast of Rat / 中国中医药信息杂志

Objective To observe the expression of core-binding factor ?1 (Cbf?1) in aging osteoblast, and try to provide new mechanisms of Chinese medicine on bone formation. Methods Rat osteoblast from different age donors (6 months and 16 months) were isolated and cultured by the tissue-fragment-migrating-growth method. The biological characteristic was observed by phase-contrast microscope and identified with mineralizing nodule staining. The expression of Cbf?1 was examined by RT-PCR and immunohistochemistry. Results RT-PCR and immunohistochemistry showed that the expression of Cbf?1 was upregulated by Gubenzhuanggu and Jinkuishenqi, and downregulated by Zhibaidihuang in aging osteoblast, but the effect was different on the Cbf?1 expression in Bushenyijing. Conclusion Buyang Chinese medicine among Bushen Chinese medicine promoted bone formation.

Pathogenic factors of blood circulation disturbance in lumbar intervertebral disc herniation and mechanism of Tuina manipulation in promoting circulation / 中西医结合学报

Tuina manipulation has long been used in the treatment of lumbar intervertebral disc herniation, although its therapeutic mechanisms remain uncertain. The specific characteristic syndrome of this disease is lumbocrural pain in varying degrees, which is due to mechanical compression, aseptic inflammation, blood circulation disturbance and dystrophy. These factors can act on the involved nerve roots and give rise to nerve conduction disturbance. Since the blood circulation disturbance and nerve roots dystrophy are the most important pathogenic aspects of this disease, the therapeutic effect of Tuina manipulation on lumbar intervertebral disc herniation is related to them directly or indirectly.

Effect of Ovariectomized Rat Serum Containing Bushen Yijing Decoction on the Absorption Function of Osteoclast / 中药新药与临床药理

Objective: To observe the effect of ovariectomized rat serum containing Bushen Yijing decoction (BYD) on the absorption function of osteoclast. Method: Fourty 10 - month - old female SD rats were randomly allocated to four groups, among which 30 rats were subjected to ovariectomy to establish the osteoporosis model, and 10 rats only received mimic operation. After 3 months, the model rats were given saline, BYD and estrogen respectively. Three months later, all the rats were killed to obtain the sera. Osteoclasts were isolated from the extremities of 1 - day SD rats and incultured on the ivory slices. The lacunar number formed after bone absorption was observed. Results: Compared with the mimic operation group, the number of lacurae of ovariectomized groups increased by 74. 1 % ,while that of the BYD group and estrogen group decreased by 65.7% and 59.3% respectively, showing significant differences with the mimic group (P

Cytologic Study on Prevention and Treatment of Primary Osteoporosis with Chinese Drugs for Tonifying the Kidney / 中医杂志

In order to compare differences of effects of complex prescription for reinforcing the kidney on osteoblast (OB) and osteoclast (OC), MTT method was used to determine proliferous capability of OB, bone lacuna count was used for quantitative assessment of absorbent activity of OC. Results showed that the mean influencing rate of Jin Gui Shen Qi Pill, Bu Shen Yi Jing Prescription, Zhi Bai Di Huang Pill, and Western medicine Premarin tablet for proliferous capability of OB was 159. 2%, 121. 5%, 21. 6% and 75. 7% respectively; the mean influencing rate for bone lacuna count was 50. 2%, 21. 0%, 19. 8% and 38. 1% respectively, suggesting that the drug for even reinforcing Yin and Yang can improve proliferation of OB and inhibit activity of OC, and increase of the drug for warming Yang can strengthen the promoting action on proliferation of OB, but increase of the drug for nourishing Yin had opposed action, the drugs for nourishing Yin plus the drugs for purging the pathologic fire can inhibit activity of OC and proliferation of OB.

Effects of Different Extraction Methods on Chemical Component Contents and Pharmacological Actions of Yangxue Ruanjian Prescription / 中药新药与临床药理

Objective To establish a proper extraction method for Yangxue Ruanjian Prescription (YRP) by comparing the chemical component contents and analgesic and anti - inflammatory effect of extracts extracted from YRP by different methods. Methods Water - decocting, alcohol - refluxing and alcohol - percolating methods were adopted and component indexes of paeoniflorin and gentiopicrin were determined by HPLC. The anti - inflammatory effect of the extracts were observed on mice with xylol - induced auricular swelling and analgesic effect by hot - plate method. Results The content of gentiopicrin is similar in the extracts extracted by three methods and the content of paeoniflorin is higher in alcohol - percolating extract than in alcohol - refluxing extract, lowest in water - extraction. The analgesic effect of water - decocting extract is better than that of alcohol - refluxing extract, and that of alcohol - percolating extract is weakest; the anti - inflammatory effect of the three extracts is almost the same. Conclusion Water - decocting extraction is suitable to YRP, and this is determined by the characteristics of Chinese compound prescription.

Fingerprint comparison of the different extraction of Bushen recipe made by decocting together and single / 中成药

AIM: To study on the differences of chemical compos i tion between extracts of Bushen recipe prepared by decocting together and single , with fingerprint technology. METHODS: The extracts of Bushen recipe were prepared by decocting together and single under the same process. The comparison was done under the different wavelength s and different brand columns, HPLC method determined the extract's fingerprint within the range of optimized conditions. RESULTS: The ratio of The compositions and its main chemical substa nces are the different between the two extracts, and the extract rate of the main chemical substances are higher in extrat decocted single than in extract decocted togethe r. CONCLUSION: There are some chemical substances and composition's ra tio differences in the extracts of Bushen recipe made by decocting together and single.