Development and construction of comprehensive examination database for periodontology based on curriculum integration / 中华医学教育探索杂志

Periodontology is one of the important disciplines in oral clinical medicine, which covers a wide range of subjects and intersects with many basic disciplines. Under the environment of the implementation of modular teaching in Shanghai Jiao Tong University School of Medicine, the assessment method with separate propositions for the teaching and research section is still adopted. There is a mismatch between the assessment mode and the curriculum setting; the basic subject propositions are difficult to be combined with clinical cases; the knowledge point assessment is single, and the students' ability to integrate the knowledge points cannot be assessed. The development and construction of the comprehensive examination database for periodontology was based on curriculum integration, gathering the teaching backbones of various disciplines, focusing on periodontology, radiating all related disciplines, unifying the proposition outline, proposition type, proposition principle, combining with relevant knowledge points of various disciplines based on clinical cases, and tried to apply to clinical students majoring in stomatology. The use of the examination database promotes students' ability to flexibly apply theoretical knowledge to clinical case analysis, further promotes the reform of modular teaching, lays a solid foundation for future clinical work, and meanwhile provides an important basis for directions of the teaching and research section.

Expression profile of long non-coding RNA in gingival tissues from the patients with aggressive periodontitis / 中华口腔医学杂志

Objective@#To detect and analyze the differential expression profile of long non-coding RNA (lncRNA) in aggressive periodontitis (AgP) and healthy gingival tissues, in order to explore the role of lncRNA in AgP.@*Methods@#After the informed consents were obtained, gingival tissues from AgP patients (n=40) and healthy volunteers (n=40) were collected in Department of Periodontology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine (from Mar. 2012 to Aug. 2012) and Department of Periodontology, Hospital of Stomatology, Tianjin Medical University (from Oct. 2016 to Apr. 2017). The differential expression of lncRNA of tissues from AgP patients (n=20) and healthy volunteers (n=20) were examined via microarray assay. Bioinformatics was applied to analyze the expression data of lncRNA and correlative mRNA. Two lncRNAs (lncRNA-TNFRSF13C and lncRNA-API5) were chosen to verify the microarray results by using real time quantitative polymerase chain reaction (PCR) in the other gingival tissues.@*Results@#Compared with the result of healthy gingival tissues, totally 8 632 lncRNAs were differentially expressed in tissues from AgP patients. From these data, 1 986 lncRNAs were significantly upregulated while 6 646 lncRNAs were downregulated, amongst which 48 lncRNAs were upregulated (>10 times) (P<0.05), 14 lncRNAs were downregulated (>10 times) (P<0.05). Furthermore, totally 5 519 correlative mRNAs were differentially expressed, amongst which 1 676 mRNAs were upregulated (≥2 times, P<0.05) and 3 843 mRNAs were downregulated≤0.5 (P<0.05). The selected lncRNA-TNFRSF13C and lncRNA-API5 were up-regulated in AgP (P<0.05), which confirmed the results of microarray. From bioinformatics, differential expression lncRNAs were in association with many signal pathways including toll-like receptor signaling pathway, cell cycle and apoptosis pathway, and tumor necrosis factor receptor superfamily pathway.@*Conclusions@#LncRNA may be involved in the pathogenesis of AgP through various pathways, which need to be further explored.

Analysis of 3-year survival of implants in patients with chronic periodontitis / 上海交通大学学报（医学版）

Objective · To observe short and medium term survival of implants in patients with chronic periodontitis after implantation. Methods · 54 patients with chronic periodontitis (133 implants) were enrolled from August 2011 to August 2013. The survival of implants was observed and the periimplant diseases were compared and analyzed between patients with different degrees of chronic periodontitis. Results · The 3-year survival rate of implants was 97.74%. The differences between patients with different degrees of chronic periodontitis were not statistically significant (P=0.452). Periodontal pocket depth (PPD) and modified plaque index (mPLI) were significantly higher in patients with severe chronic periodontitis than in patients with mild and moderate chronic periodontitis. For patients not receiving supportive periodontal therapy (SPT), the peri-implantitis rate in patients with severe chronic periodontitis was significantly higher than that in patients with mild and moderate chronic periodontitis (P=0.009). For smokers, the periimplantitis rate in patients with severe chronic periodontitis was significantly higher than that in patients with mild and moderate chronic periodontitis (P=0.016). Conclusion · For patients with chronic periodontitis, the theraputic effect of implant treatment is good. Plaque control, SPT, and smoking cessation can reduce the incidence of peri-implantitis.

Epidemiological study on periodontal health status of medical students / 上海交通大学学报（医学版）

Objective·To investigate and analyze periodontal health knowledge and behaviors in medical students and the relationship of these knowledge and behaviors with periodontal health status, and to determine the periodontal health level of this population. Methods·A total of 602 medical students were included in this cross-sectional epidemiological study. The questionnaire covered sociodemographic information, periodontal health-related behaviors and knowledge, experience about themselves, and periodontal health of parents, etc. Meanwhile, periodontal health indices of index teeth were examined, including probing depth (PD), clinical attachment loss (CAL), and bleeding on probing (BOP), etc. Results·Of 570 subjects aged 16-26 who completed the survey, 79.82% never used dental floss, and 78.25% never underwent periodontal debridement. 50.25% of the index teeth had BOP, and only 0.70% of the subjects had no BOP. 81.05% of the subjects had some degree of periodontal attachment loss. Male students were more susceptible to periodontitis (P=0.027) and gingivitis (P=0.012) than female students. Conclusion·No new risk factors affecting the periodontal health are identified. Regular periodontal cleaning and protection are important for young people to prevent periodontitis.

Establishment of a replicative senescence model of human gingival fibroblasts / 上海交通大学学报（医学版）

Objective · To establish a replicative senescence model of human gingival fibroblasts (hGFs), investigate changes in aging related biological characteristics, and provide an efficient cell model for further study on the aging in periodontal diseases. Methods · hGFs were isolated from healthy gingival tissues and cultured with tissue block method in vitro. The tissue source was verified with immunofluorescence. hGFs were continuously cultured and cumulative population doublings (CPD) were calculated and used to draw the curves. Changes in the proliferative capacity of hGFs with CPDs of 10.82, 20.65, 29.52, 42.22, 60.79, and 70.03 were examined with CCK-8. Real-time PCR was used to evaluate changes in the mRNA expression of senescence-related genes p16INK4a and p21Cip1. Results · CPD curves showed that after continuous culture, the CPD value increased gradually and became stable after achieving 70.03. hGFs became flatter and more cell rocessesappeared with the increase of CPD value. The cell proliferative capacity declined and mRNA levels of p16INK4a and p21Cip1 significantly increased (P=0.000). Conclusion · A replicative senescence model of hGFs is established throughcontinuous culture. CPD curves can reflect the aging of hGFs.

Clinical outcomes of a modified laterally moved and coronally advanced flap combined with a connective tissue graft for the treatment of severe recession defects / 上海交通大学学报（医学版）

Objective · To observe the clinical outcomes of a modified laterally moved and coronally advanced flap combined with a connective tissue graft (CTG) for the treatment of severe Miller class Ⅱ or class Ⅲ isolated recession defects. Methods · Three patients with initial defect depths of more than 5 mm and malposition in some teeth were enrolled and underwent a modified laterally moved and coronally advanced flap combined with CTG. Recession depth (RD), keratinized tissue height (KTH) of both donor and adopted site, pocket depth (PD), and clinical attachment loss (CAL) at baseline and follow-up one-year after treatment were documented. Root coverage rate (RC) was calculated and visual analogue scale (VAS) was used to evaluate patient's satisfaction degree. Results · The mean RDs at baseline and followup were (5.3±0.5) mm and (0.3±0.5) mm. The mean RC at follow-up was (93.3±9.4)% and two cases had complete root coverage. The KTHs at adopted and donor sites were (0.3±0.5) mm and (6.0±0.8) mm at baseline and (4.3±0.5) mm and (5.7±1.3) mm at follow-up, respectively. PD and CAL were decreased from (1.7±0.5) mm and (7.0±0.8) mm at baseline to (1.3±0.5) mm and (1.3±1.2) mm at follow-up, respectively. The VAS value was 9.0±0.8 and subjective evaluation of patients was improved significantly at one-year follow-up, including root sensitivity and aesthetics. Conclusion · The modified laterally moved and coronally advanced flap with CTG has ideal clinical outcomes and satisfaction degree for the treatment of patients with severe recession defects that lack keratinized tissue and combine with buccal malposition.

Application of CBCT in the detection of gingival biotype of Han nationality youths in Shanghai / 上海交通大学学报（医学版）

Objective · To detect gingival thickness of the anterior teeth region of Han nationality youths in Shanghai by cone-beam computerized tomography (CBCT), and evaluate its clinical application feasibility and the gingival biotype. Methods · Firstly, gingival thickness in the same site (5 participators, 30 sites) was detected by bone sounding and CBCT respectively, and the data were compared. A total of 30 participators with healthy gingival were recruited to the study and examined by the CBCT, the gingival thickness of selected sites (330 sites) was assessed and compared. All the subjects were examined by the experienced doctors and classified into three groups, thick-type middle-type and thin-type. Gingival thickness range and the proportion of every type were obtained. All data analyses were performed using SPSS 13.0. Results · There was no statistical difference in the thickness of gingival measured by bone sounding and CBCT (P>0.05). The main gingival biotypes of Han nationality youths in Shanghai were thin-type and middle-type. The average gingival thickness of upper central incisors [(1.32±0.15) mm] was larger than those of upper lateral incisors [(1.07±0.16) mm,P=0.000] and upper canines [(1.08±0.18) mm, P=0.000]. Conclusion · CBCT is feasible for detecting gingival thickness. Gingival thickness of the upper central incisors is significantly larger than those of upper lateral incisors and upper canines. The main gingival biotype of Han nationality youths in Shanghai is middle-type, the proportion of thick-type is least.

Clinical Study on Acupuncture Combined with Back Shu Acupoint Catgut Embedding Therapy in Treating Allergic Rhinitis / 中国中医药信息杂志

Objective To investigate the clinical efficacy of acupuncture combined with back Shu acupoint catgut embedding therapy in treating allergic rhinitis. Methods Totally 66 allergic rhinitis patients were included. Random number table method was used to divide the 66 patients into observation group and control group, with 33 cases in each group. The control group was given Ketotifen Fumarate Nasal Drops nose drops 2 drops each time, 3 times a day, Loratadine Tablets 10 mg orally, once a day for 4 weeks. The experimental group was treated with acupuncture, once a day, 10 d as a treatment course, three courses in total; at the same time, back Shu acupoint catgut embedding therapy was given, 15 d each time, twice treatment in total. After the end of treatment, clinical efficacy, clinical symptom score, life quality score, serum levels of inflammatory factors, serum immunoglobulin, peripheral blood eosinophil count and the incidence of adverse reactionsof the two groups were observed. Results The total effective rate was 93.94％ (31/33) in observation group and 75.76％ (25/33) in the control group, with statistical significance in the two groups (P0.05). Conclusion Acupuncture combined with catgut embedding therapy in the treatment of allergic rhinitis has a significant clinical efficacy, which can improve the level of inflammatory factors in patients with high safety.

Discussion of the system for full-period personnel cultivation of new nurses in Chinese medicine hospitals based on Fink curriculum construction / 中国实用护理杂志

Objective To build an efficient, scientific system for full-period personnel cultivation of new nurses in Chinese medicine hospitals. Methods This article used Fink curriculum model as the research method; used Broome′s classification theory of teaching goal as the theoretical guidance; used 'the training outline for new nurse (trying out)' issued by national health and family planning commission, the book 'the core competencies of nursing for traditional Chinese medicine', and cored competencies of nurses as the framework; used Delphi method and Analytic hierarchy process as research tools, so as to help the establishment of system for full-period personnel cultivation of new nurses in Chinese medicine hospitals. Results The preliminary exploration of system for full-period personnel cultivation of new nurses in Chinese medicine hospitals based on Fink curriculum, to make the system became more scientific, reasonable and comprehensive. Conclusions The system for full-period personnel cultivation of new nurses in Chinese medicine hospitals is feasible and creative, it can help to motivate the implementation of 'the training outline for new nurse (trying out)', to improve the quality of the nursing talents, to fasten the construction of nursing talents, to promote the development of nursing career.

Academic origin, development and characteristic of Xujiang acupuncture school / 中国针灸

The origin time, representative physicians and medical works of Xujiang acupuncture school were traced, so as to explore the academic origin and development and summarize the academic characteristic of Xujiang acupuncture school, which could make a better inheritance of academic essence and prompt the innovation and development of Xujiang acupuncture school.

Comparison study of infantile tuina between LI Dexiu and LIU Kaiyun / 中国针灸

With esoterica of Infantile Tuina by LI Dexiu and Infantile Tuina Therapy of LIU Kaiyun, the differences and similarities of manipulations, acupoints and the principles of treatment were studied so as to provide theoretical evidence to popularize tuina of LI Dexiu and LIU Kaiyun.

Risk factors related to early survival after lung transplantation for idiopathic pulmonary fibrosis / 中华器官移植杂志

Objective To estimate the effects of risk factors on early survuval (90 days) after lung transplantation for idiopathic pulmonary fibrosis (IPF).Method We reviewed 49 cases of lung tansplant male patients which suffered from IPF.Two groups were set up according to the early survival.The early outcomes (90 days) were compared between two groups by multiple logistic regression analysis.Result The early survival rate was 81.6％.Multivariate analysis confirmed that mean pulmonary artery pressure and bilateral lung transplantation (BLTx) were risk factors after adjustment for potential confounders.Recipients' age,lung volume reduction on donors,and use of extracorporeal membrane oxygenation (ECMO) were not risk factors for early mortality.Conclusion The increased pulmonary artery pressure and BLTx are risk factors for death after lung transplantation in IPF.Preoperative evaluation of mean pulmonary artery pressure and choosing suitable operative method could improve the surgical outcomes of lung transplantation.

Construction of recombinant adenovirus vector expressing hIL-17F and functional study of expressed IL-17 F / 中华微生物学和免疫学杂志

Objective To construct a recombinant adenovirus vector ( Ad-hIL-17F) expressing human interleukin 17F (hIL-17F) and to investigate the effects of expressed hIL-17F on angiogenesis. Methods The hIL-17F fragments was amplified by PCR using pUCm-T/hIL-17F plasmids as templates and then cloned into pAdTrack-CMV transfer vector to form pAdTrack-CMV-hIL-17F.The pAdTrack-CMV-hIL-17F transfer vector was linearized with PmeI digestion and then transformed into competent BJ 5183 with pAdEasy-1 backbone vector for homologous recombination .Then it was linearized with PacI digestion and transfected into human embryonic kidney 293 (QBI-293A) cells to construct Ad-hIL-17F.RT-PCR analysis and indirect immunofluorescent assay (IFA) were performed to determine the expressions of hIL-17F.MTT assay was used to detect the inhibitory effects on cell growth of ECV 304 .The expressions of VEGF and Ang-1 in 293 A and ECV304 cells were measured by ELISA .The effects of Ad-hIL-17 F on the expressions of VEGF in 293A cells were analyzed by Real-Time PCR.Results The sequencing result verified that hIL-17F gene fragment was correctly inserted in the vector .The expressions of hIL-17F gene at mRNA and pro-tein levels were confirmed by RT-PCR and IFA.Ad-hIL-17F could significantly inhibit the growth of ECV304 cells and down-regulate the expressions of VEGF and Ang-1 in 293A and ECV304 cells.Conclu-sion Ad-hIL17F expressing hIL-17F was successfully constructed .The expressed hIL-17F could inhibit the angiogenesis through down-regulating the expressions of VEGF and Ang-1.

Effect of adenovirus-mediated ING4 and IL-24 co-expression on chemosensitivity to human lung adenocarcinoma in vitro and in vivo / 生物工程学报

To study the chemosensitivity and the mechanisms of recombinant adenovirus vector expressing ING4 and IL-24 (Ad-ING4-IL-24) on lung adenocarcinoma in vitro and in vivo, the expression of ING4 and IL-24 in A549 cells was detected by RT-PCR and Western blotting. The growth inhibition, apoptosis rate and apoptosis body were measured by MTT, flow cytometry and Hoechst staining respectively. For in vivo study, we first established the A549 tumor model by grafting A549 cells in athymic nude mice; and then injected Ad-ING4-IL-24 into the tumors. Two weeks after injection, we killed the mice, removed the tumors, weighted and calculated the ratios of tumor-suppression. We also detected the expressions of ING4, IL-24, bax, bcl-2, VEGF with immunohistochemistry. The results indicated that ING4 and IL-24 were proved successfully transcription and expression in A549 cells. More interestingly, the joint group inhibited the growth of A549 cells and induced apoptosis. The in vivo data showed that the joint group suppressed the tumor growth conspicuously through up-regulating the expression of bax, and down-regulating the expression of bcl-2, VEGF. The study proved that Ad-ING4-IL-24 significantly enhanced the chemosensitivity to anticancer drug DDP in lung adenocarcinoma, which may related with cell apoptosis and antiangiogenesis.

The anti-tumor effect by adenovirus-mediated ING4 and IL-24 co-expression on hepatocellular carcinoma in vitro / 中华微生物学和免疫学杂志

Objective To construct a recombinant adenoviral vector carrying and co-expressing human inhibitor of growth 4(ING4) and human interleukin-24(IL-24) mediated by poly( A)-Promoter[Ad-ING4-poly(A)-Promoter-IL-24, referred to as Ad-ING4-IL-24] and explore its effect on the growth of HepG-2 human hepatocellular carcinoma cellsin vitro. Methods The poly(A)-Promoter(hEFl-elF4g) (Sal Ⅰ and Not Ⅰ ), ING4 ( Bgl Ⅱ and Sal Ⅰ ), and IL-24 ( Xho Ⅰ and Xba Ⅰ ) fragments were amplified by PCRusing pORF-mbcl-2α, pcDNA3.0-IL-24, and pcDNA3.0-ING4 plasmids as templates and subcloned into pAdTrack-CMV transfer vector to form pAdTrack-CMV-ING4-poly (A)-Promoter-lL-24, respectively. The pAdTrack-CMV-ING4-poly (A)-Promoter-IL-24 transfer vector linearized with Pme Ⅰ digestion and pAdEasy-1 backbone vector was further cotransformed into the bacteria BJ5183 competent cells for homologous recombination. The resultant pAdEasy-l-pAdTrack-CMV-ING4-poly ( A )-Promoter-IL-24 homologous recombinant plasmids were linearized with Pac Ⅰ digestion and transfected into the human embryonic kidney 293 (QBI-293A) cells by liposome, leading to formation of the recombinant adenoviruses Ad-ING4-IL-24co-expressing ING4 and IL-24. The Ad-ING4-IL-24 were amplified in QBI-293A cells and its titer was up to 3.5 × 109 PFU/ml. Adenovirus-mediated ING4 and IL-24 genes expression in HepG-2 cells was examined by RT-PCR and Western blot. The growth-suppressing and apoptosis-inducingg effect of Ad-ING4-IL-24 coexpressing ING4 and IL-24 on HepG-2 human hepatocellular carcinoma cells was assessed by MTT assay and FCM, respectively. Results DNA sequencing showed that the ING4, poly (A)-Promoter, and IL-24 fragments subcloned into pAdTrack-CMV plasmids were completely identical to those reported in GenBank.ING4 and IL-24 gene mediated by adenovirus could both successfully express in HepG-2 cells. Adenovirusmediated ING4 and IL-24 co-expression significantly suppressed HepG-2 hepatocellular carcinoma cell growth and induced cell apoptosis, and the effect of Ad-ING4-IL-24 group was more significant than AdING4 and Ad-IL-24 group. Conclusion The adenoviral vector co-expressing ING4 and IL-24 mediated by poly(A)-Promoter(Ad-ING4-IL-24) was successfully constructed. Ad-ING4-IL-24 had marked anti-tumor effect in suppressing HepG-2 human hepatocellular carcinoma cell growth and inducing cell apoptosis in vitro. Compared with Ad-ING4 and Ad- IL-24, Ad-ING4-IL-24 enhanced anti-tumor effect.

The effect of adenovirus-mediated ING4 expression on breast cancer cells / 中华普通外科杂志

Objective To investigate the inhibitory effect of inhibitor 4 of growth(ING4)delivered by adenovirus on human breast carcinoma cell MDA-MB-231.Methods MDA-MB-231 human breast carcinoma cells were irfected with Ad-ING4.The expression level of ING4 gene was detected by RT-PCR and Western blot;The growth inhibition,cell-cycle alteration,and apoptosis were detected by MTT,Flowcytometry and Hochest33258 staining.respectively.RT-PCR was used to detect the transcription of Bax,Bc1-2,Survivin genes;The expression level of Ang-1 gene was detected by ELISA;Ad-ING4 was given intratumorally in athymic nude mice beating MDA-MB.231 tumors.and then tumor growth was monitored;The expression of Bc1-2,Bax and Caspase-3 was analyzed by immumohistochemistry.Results ING4 was successfully transcribed and translated iU MDA-MB-231 cells:Ad-ING4 significantly inhibited the proliferation and induced G_2/M phase arrest to(24.86±1.24)% and cell apoptosis of MDAMB-231.Intratumoral injection of Ad-ING4 suppressed the tumor growth obviously with a inhibitory rate of 49%;Immumohistochemistry showed that the expression of Bax,Caspase-3 were up-regulated and the expression of Bc1-2.Survivin,CD34 were down-regulated by Ad-ING4.Conclusions Ad-ING4 can inhibit the growth of MDA-MB-231 cells and induce apoptosis in vitro and in vivo.

Construction of adenovirus vector expressing IL-24 and E1A and its inhibition of SMMC-7721 / 生物工程学报

We constructed the recombinant adenovirus vector expressing IL-24 and E1A (Ad-IL-24-E1A) and investigated the inhibition of Ad-IL-24-E1A on SMMC-7721 hepatocellular carcinoma in vitro. We amplified IL-24 gene by PCR using pAdTrack-IL-24 as template. The IL-24 gene was cloned into pAdTrack-IRES at the Bgl II and Sal I site to form pAdTrack-IL-24-IRES. E1A digested from pAdTrack-E1A was cloned into the pAdTrack-IL-24-IRES at the Xho I and EcoR V site to form the pAdTrack-IL-24-IRES-E1A. We co-transformed both pAdTrack-IL-24-IRES-E1A and pAdeasy-1 digested by Pme I and packaged to obtain Ad-IL-24-E1A. Ad-IL-24-E1A at 50 MOI infected SMMC-7721 cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay determined cell proliferation. Flow cytometry detected Cell apoptosis. The apoptotic rate of SMMC-7721 cells was 52% 48 h after infection with Ad-IL-24-E1A. The result showed that the growth of SMMC-7721 cells was significantly inhibited by Ad-IL-24-E1A at the MOI of 50.

Adenovirus mediated IL-24 gene expression inhibits growth of human glioma cell in vitro / 生物工程学报

To investigate the inhibitory effect and anti-cancer mechanism of adenovirus mediated IL-24 gene expression on the human U251 glioma cell. U251 glioma cells were infected with Ad-IL-24 at various multiplicity of infection (MOIs). Cell proliferation was determined by MTT assay. Cell apoptosis was detected by flow cytometry and Hochest staining. The transcription of apoptosis-related genes was analyzed by reverse transcription-PCR (RT-PCR), and the expression of Cleaved Caspase-3 was analyzed by Western blotting. The result showed that the growth of U251 glioma cells was significantly inhibited by Ad-IL-24 at the MOI of 100. The apoptotic rate of U251 glioma cells was 42% 72 h after infection with Ad-IL-24. Four days after infection, the growth of the U251 glioma cells was inhibited to 50%. RT-PCR showed that Ad-IL-24 not only up-regulated expression of bax/bcl-2, ICE, C-myc, p53 and down-regulated the expression of HIF-1alpha, but also enhanced Caspase-3 activation, eventually resulting apoptosis. Taken together, these results suggest that infection of U251 glioma cells with Ad-IL-24 can inhibit growth and induce apoptosis significantly by the regulation of apoptosis-related genes.

Effect of regenerated silk fibroin film on transcription and expression of vascular endothelial growth factor gene / 生物医学工程学杂志

As a biomaterial to be used for reparation in the case of trauma, the silk fibroin, particularly its effect on the transcription and expression of VEGF gene, is a concern. In this study, the ECV304 cell's growth shape and growth curve on the regenerated silk fibroin film were observed, and its VEGF secretion level was measured by ELISA test. It was found that the regenerated silk fibroin film did not interfere with ECV304 cell's growth and function. The L929 cell transfected with human VEGF gene grew on the regenerated silk fibroin film; the real-time quantitative RT-PCR method and ELISA test were used for detecting the transcription and expression of VEGF gene. The results showed the regenerated silk fibroin film did not interfere with the transcription and expression of VEGF gene. Therefore, the regenerated silk fibroin film is a safe biomaterial for inducing vascularization with no untoward effect on the reparation of trauma.

Effects of regenerated silk fibroi film on cytokine expression in the Ad-VEGF165 transgenic fibroblast / 中国组织工程研究

BACKGROUND:Previous studies have demonstrated that regenerated silk fibroin film can promote pcDNA3.0-vascular endothelial growth factor 165(rEGF165) transfected L929 cells to express VEGF.OBJECTIVE:To investigate the effects of regenerated silk fibroin film on expression of cytokines related to angiogenesis in the fibroblasts transfected by adenovirus mediated-VEGF165(Ad-VEGF165).DESIGN,TIME AND SETTING:ontrolled observational cell gene engineering experiment performed by analysis of variance at the Laboratory of Cellular and Molecular Biology,Soochow University between November 2007 and ApriJ 2008.MATERIALS:Regenerated silk fibroin film was provided by Professor Li Ming-zhong,who was from Department of Material Science and Engineering,Soochow University.METHODS:The QBI-293A and WI-38 fibroblasts cultured on the regenerated silk fibroin film.polyvinyl chloride film and (Ad-GFP)and treated by phosphate buffered saline(PBS)for controls.MAlN OUTCOME MEASURES:VEGF mRNA was detected by real-time reverse transcription-polymerase chain reaction (RT-PCR);the expression levels of VEGF,angiogenin 1(Ang 1),fibroblast growth factor 2(FGF2),and platelet-derived growth factor(PDGF)were detected by enzyme-labeled immunosorbent assay(ELISA).RESULTS:The VEGF mRNA expression in the fibroblasts cultured on the regenerated silk fibroin film was increased but that in the fibroblasts cultured on the polyvinyl chloride film was signifcantly decreased(P<0.05).ELISA results demonstrated that not only VEGF gene expression in 293A and WI-38 cells transfected bv Ad-VEGF165 cultured on regenerated silk fibroin film was high,but also Ang 1 expression increased significantly(P<0.05).Meanwhile,the expression levels of FGF2 and PDGF were normalin the fibroblasts cultured on the regenerated silk fibroin film.CONCLUSION:Adenovirus vector can be effciently transfected into fibroblasts cultured on the regenerated silk fibroin film and can express VEGF and Ang 1 protein with highly biological activity,which accelerates angiogenesis.Regenerated silk fibroin film also can maintain the normal expression levels of FGF2 and PDGF,which ale related to wound healing.