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

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 explore the bacterial flora distribution characteristics and drug resistance of pus bacterial culture in a‐cute mastitis and to analyze thechange trend of drug resistance spectrum to provide a evidence‐based basis for the rational use of an‐timicrobial agents in clinic .Methods The pus collected from 207 cases of acute mastitis was conducted the bacterial culture .The bacterial identification and antibacterial susceptibility test were performed by adopting the manual experiment combined with the DL‐96 system .Partial drug susceptibility test was performed by combining with the K‐B method .Results Among 207 specimens , 82 strains of pathogenic bacteria were detected with the detection rate of 39 .6% ,including 51 strains (62 .2% ) of staphylococcus aureus ,7 strains (8 .5% ) of pseudomonas aeruginosa ,4 strains (4 .9% ) of staphylococcus intermedius ,4 strains (4 .9% ) of staphy‐lococcus epidermis ,3 strains (3 .7% ) of acid‐producing klebsiella bacteria and each 1 strain of staphylococcus hemolyticus and other 13 kinds of bacterium .The resistance rates of staphylococcus aureus to azithromycin ,erythromycin and clarithromycin were 92 .2% ,84 .3% and 84 .3% respectively ,indicating that macrolides drugs had a higher overall drug resistance rate and were not suitable for selection and use;the resistance rates of moxifloxacin and ciprofloxacin were 3 .9% and 4 .1% respectively ,the MRSA detection rate was 27 .5% .The resistance rates of Pseudomonas aeruginosa to ticarcillin/clavulanic acid was 85 .7% ;the drug resitance rate of cefoperazone was 83 .3% ;which of gentamycin and amikacin was 71 .4% ;which of aztreonam was 14 .3% ;which of ceftazidime was 28 .6% and which of meropenem was 28 .6% .Conclusion The majority of detected bacteria in pus from the pa‐tients with acute mastitis are Staphylococcus aureus ,followed by pseudomonas aeruginosa ,which is different from that reported by other literatures ,showing the bacterial distribution has regional difference .Staphylococcus aureus has high resistance rate to macrol‐ides antibacterial drugs ,but is highly sensitive to ciprofloxacin and moxifloxacin;Pseudomonas aeruginosa has higher resistant rate to ticarcillin/clavulanic and cefoperazone ,but it is highly sensitive to aztreonam ,ceftazidime and meropenem .Empirical medication should be comprehensively considered by combining with drug resistance spectrum of Staphylococcus aureus and Pseudomonas aeruginosa ,and the sensitive drugs should be selected according to the drug susceptibility results after the antimicrobial susceptibili‐ty test for conducting the targeted medication .

Recently,zebrafish,as a new model species,has been used widely in the study of developmental mechanism of the embryo,a model of human disease and the drug screening.Zebrafish has been applied widely in the study of the drug for tumor antiangiogenesis with the development of the advanced technology of the mutagenesis and the confocal microscopy using for observation.Zebrafish applied in the screening of tumor antiangiogenesis drug and the mechanism of tumor angiogenesis are summered.

Objective To evaluate the effect of hyperthermic intraperitoneal perfusion on blood coagulation in patients undergoing laparoscope-assisted radical surgery for gastric cancer.Methods Forty patients of both sexes, aged 40-60 yr, weighing 50-80 kg, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ , undergoing elective radical surgery for gastric cancer, were equally randomized into control group (group C) and hyperthermic intraperitoneal perfusion group (group HIP).The patients were treated with hyperthermic intraperitoneal perfusion for 1 h after the end of the radical surgery in group R.Before induction of anesthesia (T0) , immediately before hyperthermic intraperitoneal perfusion (T1), and at 1 h of hyperthermic intraperitoneal perfusion (T2) , venous blood samples were collected, blood coagulation was measured using thromboelastography, and the reaction time, coagulation time, α angle and maximal amplitude were recorded.Results Compared with group C, no significant change was found in blood coagulation parameters at T0 and T1 (P＞0.05), the reaction time and coagulation time were significantly increased, and α angle and maximal amplitude were decreased at T2 in group HIP (P＜0.05).Conclusion Hyperthermic intraperitoneal perfusion can improve blood coagulation in the patients undergoing laparoscope-assisted radical surgery for gastric cancer.

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 .