ObjectiveTo establish the human periodontal ligament fibroblasts(HPDLFs)that express BMP2 and observe their biologicl characterization. MethodsA phagemid expression vector for BMP2 (pBK-B2) was transfected into HPDLFs by using LipofectAMINE. The BMP2 expression was determined by the immunohistochemical ABC method. The alkaline phosphatase (ALP) activity, osteocalcin (OC) production and capacity of mineralization were measured in the transfected cells. ResultsBMP2 protein was expressed in HPDLFs after gene transfection. The BMP2 gene transfected cells showed prominently elevated ALP activity, OC production and increase in mineralized nodules. ConclusionThe results indicate that BMP2 is expressed in HPDLFs and is involved in inducing differ- entiation of HPDLFs into osteoblast-like cells.

Objective To investigate the effects of platelet-derived growth factor(PDGF) on the expression of ?-smooth muscle actin(?-SMA) of cultured human retinal pigment epithelium cells(RPE). Methods Cultured human RPE cells of the 4-6 th passages were divided into two groups: Delbecco′s modified Eagle′s medium (DMEM) and 2%DMEM (20 g/L foeta calf serum+DMEM). PDGF (0,1,50 ng/ml) was added to medium.The expression of ?-SMA was detected and quantitatively analyzed by image process of immunofluorescence. Results PDGF stimulated the expression of ?-SMA of human RPE cells.In group of DMEM, The rate of RPE of ?-SMA expression was 40%-50% and the intension of fluorescence was 8 08 without PDGF. After stimulated by PDGF(1 ng/ml,50 ng/ml), the rates were 80% and 90% respectively, and the intension of fluorescence were 12.35 and 17.23. In 2%DMEM group, The rates of RPE of ?-SMA expression were 85% without PDGF, and 95% ,100% respectively treated with PDGF (1 ng/ml,50 ng/ml). The intension of fluorescence was 14.79 without PDGF, and after stimulated by PDGF, they were 16.28 at 1 ng/ml and 21.36 at 50 ng/ml,which was 2.7 times stronger than that in DMEM group without PDGF. Conclusion PDGF could stimulate RPE cells to express ?-SMA.

OBJECTIVETo establish the migraine rheumatism stasis syndrome animal model.

METHODThe rat migraine rheumatism stasis syndrome animal model was established through rheumatism stimulation with manual climate box, 5-HT reduction caused by reserpine and local cerebral vasospasm. General vital signs (activity, weight, eye gum, hair, feeding, excrement), head scratch frequency and image collection were observed to analyze the changes in biological signs of stasis syndrome (tongue image RGB), thrombin and serotonin of model rats.

RESULTThe reserpine group and the reserpine plus rheumatism model group showed significant reduction in blood coagulation time, pain threshold and 5-HT content in blood and brain (P < 0.01); the reserpine plus rheumatism model group showed an increase in eye gum and decreases in activity, feeding, with thin sloppy stool. According to the tough RGB values, the control group showed light red toughs, the reserpine group showed dark purple toughs, the reserpine plus rheumatism model group showed gray toughs, with notable differences in tough RGB values in all three group.

CONCLUSIONThe rheumatism stimulation with manual climate box, 5-HT reduction caused by reserpine and local cerebral vasospasm can be used to induce the migraine rheumatism stasis syndrome animal model, but its modeling assessment method and process shall be further improved.

Animals ; Blood Circulation ; Diagnosis, Differential ; Disease Models, Animal ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Migraine Disorders ; diagnosis ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Rheumatic Diseases ; diagnosis ; physiopathology

Adult ; Heavy Metal Poisoning ; Humans ; Male ; Poisoning ; Thallium ; poisoning

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Humans ; Neuromyelitis Optica ; therapy

China ; epidemiology ; Humans ; Occupational Diseases ; epidemiology

BACKGROUND: It has been thought that posterior cervical decompression increases the risk of cervical instability and the stress on posterior longitudinal ligament may change growth factor expression in vivo. However, the association between BMP-4 expression and heterotopic bone formation has not been confirmed. Bone morphogenetic protein 4 (BMP-4) is one of factors that can lead to heterotopic bone formation alone.OBJECTIVE: To observe the effects of posterior cervical decompression on BMP-4 mRNA expression in the posterior longitudinal ligament.DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed in Taishan Medical College between January 2005 and December 2006.MATERIALS: A total of 48 adult Sprague Dawley rats of either gender and of clean grade, weighing 270-350 g, were included in this study.METHODS: All rats were randomly and evenly divided into 3 groups: experimental, sham-operated, and blank control. In the experimental group, C36 laminectomy for decompression was performed.In the sham-operated group: only skin incision was made. Rats in the blank control group received no any treatment. Four rats were allocated for each time point (1, 2, 4, and 8 weeks post-surgery) for harvesting posterior longitudinal ligament.MAIN OUTCOME MEASURES: Detection of BMP-4 mRNA expression in the cervical posterior longitudinal ligament by semi-quantitative reverse transcription-polymerase chain reaction.RESULTS: In the experimental group, obvious BMP-4 mRNA expression was found at 1-8 weeks post-surgery. While no BMP-4 mRNA expression was found at all times in the sham-operated and blank control groups.CONCLUSION: Posterior cervical decompression can increase BMP-4 mRNA expression in the posterior longitudinal ligament.Endogenous BMP-4 may contribute to ligamentous ossification and development post-surgery.

Objective To identify the expression of a fusion gene GCA formed from GM-CSF gene and LMP2A gene of Epstein-Barr virus (EBV) in a recombinant BCG (rBCG) and to study its immunoge-nicity.Methods The rBCG was constructed to express the fusion gene GCA and the expressed products were detected by Western blot assay .ELISA was performed to measure specific antibody titers in serum sam-ples from mice immunized with rBCG .Lactate dehydrogenase assay was used to analyze the cellular immuni-ty of mice.A mouse model of EBV-positive gastric carcinoma was established to evaluate the therapeutic effects of rBCG.Results The target proteins of GM-CSF and LMP2A were successfully expressed in rBCG . The specific antibodies were detected in rBCG immunized mice as indicated by ELISA .The maximum anti-body titer reached 1 ∶27 900 [(326.5±7.8) pg/ml] as injection with rBCG 5×108/mouse.The rBCG in-duced cytotoxicity of cytotoxic lymphocytes (CTLs) to EBV-positive gastric carcinoma cells (GT39) (with a killing rate of 89.6%±6.8%) was significantly higher than that of control group (P<0.05) The sizes of tumor in PBS control group [(1964.0±548.7) mm3] and BCG group [(1268.65±72.4) mm3] were big-ger than those in rBCG group [(168.64±78.80) mm3].Conclusion The rBCG expressing GM-CSF and LMP2A fusion gene was successfully constructed .The rBCG could induce humoral and cellular immune re-sponses in mice and inhibit the growth of tumor .

The construction of a network resource data base for clinical skill teaching aiming at sharing excellent teaching resources was completed by integrating all kinds of teaching resource including characters,eourseware,pictures,cartoons,videos and examination questions,etc.In view of the generally weak situation of the current domestic education resource data base construction,the contents,guiding ideology,principle,object,orientation,function and the management mechanism of the network resource data base construction were devised for clinical skills teaching.Suggestions for specific ideas and construction problems were proposed to promote the construction and optimization of the network resource database for clinical skills teaching.

Objective To compare the differences between external standard method and relative correction factor method for determination of the flavonoids from Sorbus tianschanica Rupr.Methods Using HPLC external standard method for determination of hyperoside,rutin,isoquercitrin,quercetin-3-O-(6″-O-malonyl)-β-D-glucoside,astragalin and Kaempferol-3-O-(6″-O-malonyl)-β-D-glucopyranoside in Sorbus tianschanica Rupr.,HPLC relative correction factor method was adopted to establish relative correction factor of the other five flavonoids above with hyperoside as reference.The difference was evaluated by comparing the external standard method with the relative correction factor method.Results There was no significant difference between the T test external standard method and relative correction factor method(P>0.05).Conclusion External standard method and relative correction factor method can be used for determination of the flavonoids from Sorbus tianschanica Rupr.,but in the case of lack of reference substance or mass detection,using the relative correction factor method for determination of rutin,hyperoside isoquercitrin,quercetin-3-O-(6″-O-malonyl)-β-D-glucoside,astragalin and kaempferol-3-O-(6″-O-malonyl)-β-D-glucopyranoside in Sorbus tianschanica Rupr.It was more feasible and it can be used as a new quality evaluation method in determination of flavonoid components from Sorbus tianschanica Rupr.