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.

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 .

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.

Type 2 diabetes,a glucose and lipid metabolism disorder accompanied by chronic multiple organ damage,has become a huge threat to human health,α/β hydrolase domain-6 (ABHD6) regulates the insulin release negatively by hydrolyzing monoacylglycerol.Small molecule ABHD6 inhibitors have been proven to lower bloodglucose and regulates energy homeostasis,which is a potential candidate for the treatment of type 2 diabetes.This paper introduced the ABHD6 signaling pathway and its mechanism,then reviewed the progress of small molecule ABHD6 inhibitors with different structures in recent years,and analyzed the structure activity relationship.

Objective To explore the behavior characteristics in children with benign epilepsy combined with centro-temporal spikes(BECTS).Methods Eighty-two children with BECTS aged 2.5-3.0(2.65?2.31)years old,51 male,31 female,who were free of mental retardation assessed with Gesell developmental schedules,untreated with antiepileptic drugs,and were investigated 15 days after the latest seizure.Eighty-two healthy children with sex and age matched to the cases,53 male,29 female,aged 2.5-3.0(2.6?0.4)years old.The behavior characteristics of infants in BECTS group and control group were assessed with CBCL,including 6 behavior factors which were sleep problem,social flinches,depression,physical aspect,attacking,act of sabotage and the infants-middle school student social ability scale.Results The total scores of behavior characteristics and the scores of depression,sleep problem,attacking and act of sabotage in BECTS group were all higher than those in control group,the differences were statistically significant.However,scores of social flinches,and physical aspect in BECTS group had no significant differences compared with those of control group.There were no significant difference of social adaptive component between the BECTS and control group.Conclusions Children with BECTS have behavior disorders to some extent,but their social adaptive capacity are the same as normal children.

Objective To investigate the effect of different endoscopic sclerotherapy for esophageal variceal bleed-ing caused by rupture. Methods 66 cases of liver cirrhosis with esophageal variceal bleeding were randomly divided into treatment group A, endoscopic lauromacrogol sclerotherapy group B and endoscopic aethoxysklerol 1% German agents in treatment of group C, then observe the successful hemostasis, the curative effect and follow-up of patients with varices, bleeding, fever and other adverse conditions. Results Group B and group C with success rate of hemostasis has obvious advantages compared with group A, <0.05, the difference was statistically significant;total effective rate of group B was significantly higher than that in group A, < 0.05, the difference was statistically sig-nificant; early period of rebleeding, delayed again bleeding in group B was occurred less than that of group C, >0.05, the difference was not statistically significant; adverse conditions such as fever, ulcers, pain in group B was less than that in group C, <0.05, the difference was statistically significant. Conclusion Endoscopic lauromacrogol injection sclerotherapy for esophageal variceal bleeding is a safe, fast speed, timely, effective and repeatable treat-ment, it is worthy of promoting clinical application.

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 evaluate the safety and efficacy of icotinib as first-line therapy in Chinese non-small cell lung cancer (NSCLC) patients harboring epidermal growth factor receptor (EGFR) sensitive mutations.Methods Patients with stage ⅢB/Ⅳ NSCLC who had EGFR sensitive mutation and had no previous treatment were enrolled into this study.The response rates , progress free survival ( PFS) , overall survival ( OS ) , and the safety were analyzed.Results Ninety advanced adenocarcinoma patients were enrolled in this study , 44 patients had partial response ( PR ) , 42 patients had stable disease ( SD ) , 4 patients had progressive disease (PD), with an overall response rate (ORR) of 48.9%, and a disease control rate (DCR) of 95.6%.The median PFS was 14.9 months (95%CI 13.5-16.3) and the OS was 37.0 weeks ( 95%CI 27.9 -46.1 ).Patients with brain metastases showed higher ORR ( P =0.049 ).Patients with stage ⅢB had longer PFS than those with stage Ⅳ( P=0.007 ).The most common adverse events were grade 1 -2 skin rash (38 patients, 40.9%).Other adverse events included dry skin , oral mucositis, diarrhea and liver function injury.Three patients withdrew because of severe liver injury or skin rash.No treatment related mortality occurred .Conclusions Icotinib is effective and safe as first-line treatment for Chinese advanced NSCLC patients with EGFR sensitive mutation.

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 .