1.Adenovirns construction with miceCD40- IgG2aFc-IRS-GFP fusion gene and in vitro testing
Chuanguang LI ; Yifeng FANG ; Yingxia TAN ; Zhijian SHEN ; Qiyu ZHANG
Chinese Journal of Hepatobiliary Surgery 2010;16(3):208-211
Objective To construct the adenovirus vector containing mice CD40- IgG2aFc-IRS-GFP fusion gene and detect the expression of the fusion protein in transfeete 293cells.Methods The fragments of CD40, IgG2aFc were obtained and inserted into plasmid PDC316-IgG2aFc-GFP.After being verified by sequencing, Ad5-PDC316-CD40-IgG2aFc-GFP was contransfored with the bckbone vector into 293 cells.The virus titer was detected after replicating and purifing and the expression of the fusion protein was analyzed.Results A virus and plasmid were constructed successfully.The vitro infection showed that the virus can infect cells of which the fusion protein was confirmed by fluo-rescence.The expression of the fusion protein increased with the increased time and virus concentra-tion.The protein expression stopped increasing after the virus concentration came to a certain level.Conclusion CD40, IgG2aFc fragments are correctly ligated with plasmid PDC316-IgG2aFc-GFP, and the fusion protein can be expressed in 293cells.This might lay a foundation for further studies of the expresstion of virus, immune tolerance and mechanism of liver transplantation model in rats.
2.Construction and identification of a double-regulated replicative adenovirus AdTPHre-hEndo
Yifeng FANG ; Yunfeng SHAN ; Shenmeng GAO ; Qiyu ZHANG
Chinese Journal of Hepatobiliary Surgery 2011;17(6):488-491
Objective To develop a double-regulated replicative adenovirus carrying the Human endostatin gene(hEndo). Methods The plasmid pTPHre-hEndo was constructed by gene engineering technique, carrying human endostatin gene, in which El A gene and E1B gene were driven by human telomerase reverse transcriptase (hTERT) promoter and hypoxia response element (HRE) promoter,respectively. The pTPHre-hEndo was co-transfected with pBHGE3 in 293 cells to generate recombinant adenovirus AdTPHre-hEndo. Virus titer was measured by the TCID50 method. Virus replication assay was performed to evaluate the selective replication ability of AdTPHre-hEndo. The transgene expression of endostatin was detected by ELISA assay. Results A novel gene-viral therapeutic system AdTPHre-hEndo was constructed by gene engineering technique and its titer was 3. 25 X 1010 pfu/ml.Proliferative test revealed that AdTPHre-hEndo could proliferate selectively in telomerase positive tumors. Furthermore, in comparison with non-replicative adenovirus Ad-hEndo, the transgene expression of endostatin mediated with AdTPHre-hEndo was significantly increased (P < 0. 01).Conclusion The novel gene-viral therapeutic system AdTPHre-hEndo has the capacity to replicate in pancreatic cancer cells and expresses the endostatin efficiently, and may provide a new strategy for pancreatic cancer gene therapy.
3.Therapeutic effects of replication-competent adenovirus carrying human endostatin gene on pancreatic cancer in mice
Yifeng FANG ; Yunfeng SHAN ; Dingcun LUO ; Qiyu ZHANG
Chinese Journal of Hepatobiliary Surgery 2011;17(8):660-663
Objective To establish human pancreatic cancer xenografts in nude mice, and to investigate the antitumor efficacy of human endostatin expressed by replication-competent adenovirus AdTPHre-hE in vivo. Methods Pancreatic cancer cells AsPC-1 were injected subcutaneously in BALB/c nude mice to establish the xenografts. Tumor growth was observed and measured after AdTPHre-hE treatment. Expression of endostatin was detected by ELISA assay. The tumors were harvested for pathologic examination and immunohistochemical staining. Results Tumors grew more slowly in the AdTPHre-hE group and their sizes were markedly smaller than those of the Ad-hE group (P<0.01)and control group(P<0. 01). Endostatin levels were detected in the sera of nude mice in all treated groups, and endostatin expression in AdTPHre-hE group increased with time. The endostatin level in the AdTPHre-hE treated group was much higher(P<0. 01)and increased faster than that in the Ad-hE treated group. Immunohistochemical staining for Hexon of adenovirus capsid showed more positive tumor cells in the tumor tissues treated with AdTPHre-hE. Immunohistochemical staining for FⅧ revealed a decreased microvessel density in the tumor tissues treated with AdTPHre-hE. Conclusion The replication-competent adenovirus efficiently expressed high-level endostatin and significantly inhibited tumor growth in vivo.
4.Treatment of pancreatic cancer by replicating adenovirus mediated human endostain gene in nude mice
Xueqiang WANG ; Yifeng FANG ; Heping Lü ; Yunfeng SHAN ; Qiyu ZHANG
Chinese Journal of General Surgery 2010;25(10):797-800
Objective To investigate the effect of a dual regulation of replicating adenovirus vector carrying human endostatin gene (AdTPHre-hEndo) on pancreatic cancer. Methods Human endostatin (hEndo) gene was cloned into the genome of replicating adenovirus specific for the tumor cells by virus recombination technology. The virus titer was 3.25 × 1010pfu/ml. A Balb/c nude mouse model carring sw1990 cells pancreatic cancer was established, the expression of human endostain and inhibition of tumor cells in vivo were detected. Results We successfully constructed AdTPHre-hEndo. The inhibition on pancreatic cancer cell line SW-1990 of AdTPHre-hEndo is better than Ad-hEndo (P <0. 01 ), and ONYX-015 ( P < 0. 05 ). The endostatin expression of AdTPHre-hEndo group was significantly higher than Ad-hEndo group and the control group (P < 0. 01 ). The intratumoral MVD also decreased significantly in the treated tumors(6. 8 ±2. 5 vs. 16. 0 ±4. 6、47. 2 ± 10. 0, P <0. 01 ). Conclusions The recombination adenovirus can express biologically active hEndo effectively, which results in inhibiting the growth of micro-blood vessels and proliferation slowly.
5.Application of health management among coronary heart disease patients after stent implementation
Meilan WANG ; Qiyu FANG ; Zhe JIN ; Mei SUN
Chinese Journal of Modern Nursing 2015;21(8):888-890
Objective To explore the influence of the health management among coronary heart disease (CHD) patients after stent implementation. Methods A total of 32 CHD patients with postoperative stent implementation were randomly divided into experimental group ( received 6 months health management) and control group on (received routine discharge education) average and the results of education in two groups were compared and analyzed. Results After 6 months, there were (125. 9 ± 14. 9) mmHg, (72. 8 ± 8. 4) mmHg, (5. 44 ± 0. 53)mmol/ L, (3. 79 ± 1. 11) mmol/ L, (25. 4 ± 12. 4) U/ L, (63. 8 ± 7. 6) scores in systolic pressure, diastolic pressure, fasting blood-glucose, blood fat, liver function and body quality assessment in the experimental group compared with (133. 8 ± 11. 6) mmHg, (79. 4 ± 8. 3) mmHg, (7. 46 ± 2. 98) mmol/ L, (5. 22 ± 1. 33) mmol/ L, (38. 7 ± 13. 7)U/ L, (65. 8 ± 13. 3) scores in the control group (t = 2. 186, 2. 223, 2. 635, 2. 361, 1. 972, 2. 336,respectively;P < 0. 05). Conclusions After CHD patients acquired stent implementation, the health management can effectively minimize or stabilize the indexes of patient′ s blood pressure, blood glucose, blood fat, liver function and body quality, decrease recurrence of CHD, and promote the quality of life among CHD patients after stent implementation.
6.Deformation Characteristics of Hand Movement During Grasping
Qiyu ZHU ; Zixiang TONG ; Fang YUAN ; Kang WEI ; Xinxing SHAO ; Xiaoyuan HE
Journal of Medical Biomechanics 2021;36(3):E423-E439
Objective Based on the multi-camera digital image correlation (DIC) method, the dynamic deformation characteristics of human hand during grasping were studied. Methods A continuous four-camera DIC system was established to measure surface strain of the skin on the back of the hand during grasping process, and then through the connection between skin, joints, bones and muscles, the regular pattern of muscle deformation could be known indirectly. Results Four grasping postures (medium cylinder, lateral pinch, index finger extension, power sphere) were measured. It was found that the increases of strain magnitude were different at different positions on back surface of the hand under different grasping postures, and the maximum principal strains were between 0.1 and 0.3. The movement characteristics for each muscle group of the hand under different grasping postures were obtained through analysis. Conclusions This method has the characteristics of non-contact, full field, intuitive results, which provides a new way for in vivo measurement of dynamic deformation during grasping.
7.Rare tumors: a blue ocean of investigation.
Shuhang WANG ; Peiwen MA ; Ning JIANG ; Yale JIANG ; Yue YU ; Yuan FANG ; Huilei MIAO ; Huiyao HUANG ; Qiyu TANG ; Dandan CUI ; Hong FANG ; Huishan ZHANG ; Qi FAN ; Yuning WANG ; Gang LIU ; Zicheng YU ; Qi LEI ; Ning LI
Frontiers of Medicine 2023;17(2):220-230
Advances in novel drugs, therapies, and genetic techniques have revolutionized the diagnosis and treatment of cancers, substantially improving cancer patients' prognosis. Although rare tumors account for a non-negligible number, the practice of precision medicine and development of novel therapies are largely hampered by many obstacles. Their low incidence and drastic regional disparities result in the difficulty of informative evidence-based diagnosis and subtyping. Sample exhaustion due to difficulty in diagnosis also leads to a lack of recommended therapeutic strategies in clinical guidelines, insufficient biomarkers for prognosis/efficacy, and inability to identify potential novel therapies in clinical trials. Herein, by reviewing the epidemiological data of Chinese solid tumors and publications defining rare tumors in other areas, we proposed a definition of rare tumor in China, including 515 tumor types with incidences of less than 2.5/100 000 per year. We also summarized the current diagnosis process, treatment recommendations, and global developmental progress of targeted drugs and immunotherapy agents on the status quo. Lastly, we pinpointed the current recommendation chance for patients with rare tumors to be involved in a clinical trial by NCCN. With this informative report, we aimed to raise awareness on the importance of rare tumor investigations and guarantee a bright future for rare tumor patients.
Humans
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Neoplasms/pathology*
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Biomarkers
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Prognosis
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Oceans and Seas
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China/epidemiology*