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The use of melittin to enhance transgene expression mediated by recombinant adeno-associated virus serotype 2 vectors both in vitro and in vivo

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Journal of Integrative Medicine 2023年第1期21卷 106-116页

作者： Yi-lin Xie ji-yao wang Yun He Xiao-min Yu Qing-yun Zheng Chen Ling Xi-lin Feng Li-qing Zhu State Key Laboratory of Genetic Engineering and Engineering Research Center of Gene Technology(Ministry of Education) School of Life SciencesZhongshan HospitalFudan UniversityShanghai 200438China Department of Clinical Laboratory the First Affiliated Hospital of Wenzhou Medical UniversityWenzhou 325000Zhejiang ProvinceChina Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province Wenzhou 325000Zhejiang ProvinceChina Yantai Fuheng Biological Technology Co. Ltd.Yantai 264006Shandong ProvinceChin

Objective: Melittin, a cell-penetrating peptide, improves the efficiency of many non-viral gene delivery vectors, yet its application in viral vectors has not been well studied. The non-pathogenic recombinant adeno-associated virus(rAAV) vector is an ideal in vivo gene delivery vector. However, its full potential will only be achieved after improvement of its transduction efficiency. To improve the transduction efficiency of rAAV2 vectors, we attempted to develop a melittin-based r AAV2 vector delivery strategy.Methods: The melittin peptide was inserted into the rAAV2 capsid either in the loop Ⅷ of all viral proteins(VPs) or at the N terminus of VP2. Various r AAV2-gfp or-fluc vectors were subjected to quantitative real-time polymerase chain reaction and Western blot assays to determine their titers and integrity of capsid proteins, respectively. Alternatively, the vectors based on wild-type capsid were pre-incubated with melittin, followed by transduction of cultured cells or tail vein administration of the mixture to C57BL/6 and BALB/c nude mice. In vivo bioluminescence imaging was performed to evaluate the transgene expression.Results: rAAV2 vectors with melittin peptide inserted in the loop Ⅷ of VPs had low transduction efficiency, probably due to dramatically reduced ability to bind to the target cells. Fusing the melittin peptide at the N-terminus of VP2 produced vectors without the VP2 subunit. Interestingly, among the commonly used rAAV vectors, pre-incubation of r AAV2 and rAAV6 vectors with melittin significantly enhanced their transduction efficiency in HEK293 and Huh7 cells in vitro. Melittin also had the ability to increase the rAAV2-mediated transgene expression in mouse liver in vivo. Mechanistically, melittin did not change the vector-receptor interaction. Moreover, cell counting kit-8 assays of cultured cells and serum transaminase levels indicated melittin had little cytotoxicity.Conclusion: Pre-incubation with melittin, but not insertion of melittin into the rAAV2 capsid, significantly enhanced

关键词： TAAV Melittin Capsid engineering Co-administration Transduction efficiency

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Development and preliminary verification of the evaluation system for clinical practice guidelines in China

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Chronic Diseases and Translational Medicine 2020年第2期6卷 134-139页

作者： ji-yao wang Qiang wang Xiao-Qin wang Xue-Juan jin Bo-Heng Zhang Shi-yao Chen Xue-Cheng Gao Department of Gastroenterology Zhongshan HospitalFudan UniversityShanghai 200032China Evidence-based Medicine Center Fudan UniversityShanghai 200032China Medical Management Center of the National Health and Family Planning Commission Beijing 100191China Institute of Cardiovascular Diseases Zhongshan HospitalFudan UniversityShanghai 200032China Department of Hematology Huas han HospitalFudan UniversityShanghai 200040China Liver Cancer Institute.Zhongshan Hospital Fudan UniversityShanghai 200032China

Objective:Clinical practice guidelines can improve healthcare processes and patient outcomes;however,the quality of these guidelines varies greatly in China.The aim of this study was to construct a comprehensive instrument for the appraisal of clinical practice guidelines in China(AGREE-CHINA),and to validate its reliability as a tool for helping potential guideline users in assessing guideline quality.Methods:First,an interdisciplinary working group was established for developing the methods.They also created a checklist as a tool according to the Appraisal of Guidelines,Research and EvaluationⅡ(AGREEⅡ)standards,considering the particularity of Chinese clinical practice.Next,the first draft of AGREE-China was developed by vote,modification,preliminary trial,and crossverification.To ensure the objectivity,credibility,and reproducibility of the draft assessment,all of the checklists and standards were cross-reviewed fairly widely.Fin ally,AGREE-CHINA and AGREEⅡwere used to assess the Chinese guideli nes published in the past five years,and the results were compared.Results:The presented AGREE-CHINA covered five main checkpoints(science and rigor,effectiveness and safety,economy,usability and feasibility,and conflicts of interest)with each point divided into several more specific checkpoints.Definitions and rationales for each main checkpoint appear in the Appendix.The quality ratings based on the total scores of AGREE-China and AGREEⅡwere consistent(r=0.508,P=0.020).Compared with AGREEⅡ,the study showed a higher level of interraterreliability for AGREE-CHINA overall(ICC=0.957,P<0.001).The mean time required for AGREE-CHINA was less than that for AGREEⅡ:this was approximately 30 minutes for every assessment.User satisfaction was generally high.Conclusions:This paper has presented the first edition of the AGREE-CHINA appraisal tool for clinical guidelines.It is quick and easy to use;it assesses and performs well in comparison to AGREEⅡ.This first version of AGREE-CHINA will require further development and validation

关键词： Appraisal of guidelines research and evaluation China Development Verification

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Early diagnosis and therapeutic choice of Klebsiella pneumoniae liver abscess

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Frontiers of Medicine 2010年第3期4卷 308-316页

作者： jing LI Ying FU ji-yao wang Chuan-Tao TU Xi-Zhong SHEN Lei LI Wei jiANG Department of Gastroenterology Zhongshan HospitalFudan UniversityShanghai 200032China

Nowadays,pyogenic liver abscess(PLA)is still a common and severe intra-abdominal infection,and Klebsiella pneumoniae had emerged as the most common pathogenic bacteria worldwide in the past ten years.Our study aims to achieve an early pathogenic diagnosis and rational therapy modality for Klebsiella pneumoniae liver abscess(KLA)through clinical data analysis.A total of 197 inpatients in Zhongshan Hospital,Shanghai,diagnosed as having liver abscess between March 2001 and September 2009 were enrolled.Patients with monomicrobial infection were divided into two groups:patients with K.pneumoniae liver abscess(KLA group,n=106)and those with non-Klebsiella pneumoniae liver abscess(NKLA group,n=56).A retrospective analysis was made between these two groups on the aspects of underlying diseases,clinical characteristics,laboratory data,culture results,and imaging findings.To evaluate the effects of different medical interventions,monomicrobial KLA patients were further divided into four subgroups(percutaneous liver aspiration,aspiration plus antibiotics flushing,aspiration plus retained catheter,and aspiration plus antibiotics flushing and retained catheter),and corresponding therapeutic effects were analyzed.KLA was more likely to occur in patients with coexisting diseases such as diabetes mellitus(53.77%vs 25.00%,P=0.001)and hepatic adipose infiltration(16.04%vs 5.36%,P=0.029).Compared to NKLA group,clinical characteristics including abdominal pain(40.57%vs 57.14%,P=0.044),hypodynamia(19.81%vs 46.43%,P=0.001),and hepatomegaly(4.72%vs 14.29%,P=0.033)were much milder,but with a higher fasting blood glucose level(7.84±0.36 vs 5.76±0.30,P=0.001)on admission in KLA group.In addition,KLA abscess often appeared singly in the right lobe of the liver with gas forming nature(32.88%vs 13.51%,P=0.039),unsmooth rim(71.23%vs 40.54%,P=0.002),and dynamic septum enhancement(41.10%vs 16.22%,P=0.009).Compared to mono aspiration subgroup,additional antibiotic flushing could not further improve clinical outcomes of KLA patients(P>0.05);howev

关键词： liver abscess Klebsiella pneumoniae com-puted tomography therapy

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A randomized,double-blind,placebo-controlled trial assessing the efficacy and safety of tegaserod in patients from China with chronic constipation

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World Journal of Gastroenterology 2007年第5期13卷 732-739页

作者： San-Ren Lin Mei-Yun Ke jin-Yan Luo yao-Zong Yuan ji-yao wang Shelley diTommaso Verena Walter jiaqing Huang The 3rd Hospital Beijing Medical University Beijing China Peking Union Medical College Hospital BeijingChina The 2nd Hospital of Xian jiaotong University Shanxi China Shanghai Ruijin Hospital Shanghai China Shanghai Zhongshan Hospital Shanghai China Novartis Pharma AG Basel Switzerland Novartis Pharmaceuticals Corp East HanoverNew Jersey United States

AIM: To evaluate the efficacy and safety of tegaserod, 6 mg twice daily (b.i.d.), in men and women with chronic constipation (CC) from China. METHODS: This was a multicenter, double-blind, placebo-controlled study. Following a 2-wk treatment-free baseline period, patients were randomized to receive either tegaserod (6 mg b.i.d.) or placebo (b.i.d.) for 4 wk. An analysis of covariance with repeated measures was used to determine the overall effect of treatment for the primary efficacy variable; the change from baseline in the number of complete spontaneous bowel movements (CSBMs) during the 4-wk treatment period. Secondary efficacy endpoints included other measures of response in terms of CSBMs, and patients’ daily and weekly assessment of bowel habits. Safety was also assessed, based on the incidence and severity of adverse events (AEs).RESULTS: A total of 607 patients were randomized to receive either tegaserod (n = 304) or placebo (n = 303). Tegaserod treatment resulted in a rapid and significant increase from baseline in the adjusted mean number of CSBMs per week over wk 1-4 compared with placebo (1.39 vs 0.91, P = 0.0002). A statistically significant difference in favor of tegaserod was also observed for a mean increase ≥ 1 CSBM/wk over wk 1-4 (47.7% vs 35.0%, tegaserod vs placebo, respectively, P = 0.0018) and for the absolute number of ≥ 3 CSBMs/wk over wk 1-4 (25.0% vs 14.5%, tegaserod vs placebo, respectively, P = 0.0021). Improvements in other symptoms of CC were also seen in the tegaserod group, including improved stool form and reduced straining. In addition, more patients in the tegaserod group reported satisfactory relief from their constipation symptoms. The frequency and severity of AEs was comparable between tegaserod and placebo groups, with the exception of a greater incidence of diarrhea in patients receiving tegaserod (3.6%) compared with placebo (1.7%).CONCLUSION: Tegaserod treatment improved multiple symptoms of CC and was associated with a favorable safety profile.

关键词：慢性便秘替加色罗治疗效果安全性随机双盲对照试验

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Preparation of chitosan-polyaspartic acid-5-fluorouracil nanoparticles and its anti-carcinoma effect on tumor growth in nude mice

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World Journal of Gastroenterology 2008年第22期14卷 3554-3562页

作者： Dan-Ying Zhang Xi-Zhong Shen ji-yao wang Ling Dong Yong-Li Zheng Li-Li Wu Department of Gastroenterology Zhongshan Hospital Shanghai Medical College Fudan University Shanghai 200032 China Department of Maeromoleeular Science Key Laboratory of Molecular Engineering of Polymers of Chinese Ministry of Education Fudan University 220 Handan Road Shanghai 200433 China

AIM: To prepare chitosan-polyaspartic acid-5-fluorouracil (CTS-Pasp-5Fu) nanoparticles and investigate its anti-carcinoma effect and toxicity. METHODS: CTS-Pasp-5Fu nanoparticles were synthesized by ionic gelatification. Male BABL/c nude mice were injected with SGC-7901 gastric carcinoma cell line mass to establish a human gastric carcinoma model. They were randomly allocated into 4 groups: CTS-Pasp-5Fu (containing 5-Fu 1.25 mg/kg), 5-Fu (1.25 mg/kg), CTS-Pasp and normal saline groups. Tumor weight was measured and assay of colony forming unit-granulocyte and macrophage (CFU-GM) was performed. The structural change of cells and tissues was observed and the Bax and Bcl-2 genes were detected. RESULTS: Compared with normal saline, the inhibition rates of tumor growth for the CTS-Pasp, 5-Fu and CTS-Pasp-5Fu groups were 5.58%, 58.69% and 70.82%, respectively. The tumor inhibition rates for the CTS-Pasp, 5-Fu and CTS-Pasp-5Fu groups were 5.09%, 65.3% and 72.79%, respectively. There was a significant decrease in the number of CFU-GMformation and increase of total bilirubin, and alanine aminotransferase in the 5-Fu group, but no change in those of the other three groups. There was no change in white blood cell count and creatinine among the four groups. Pathological section of liver and nephridial tissues showed that the damage in the 5-Fu group was more severe than that in the CTS-Pasp-5Fu group. 5-Fu and CTS-Pasp-5Fu groups could both down-regulate the Bcl-2 expression and up-regulate the Bax expression to different extent, and the accommodate effect of CTS-Pasp-5Fu was more obvious than 5-Fu. CONCLUSION: The tumor inhibition rate of CTS-Pasp-5Fu nanoparticles is much higher than that of 5-Fu alone.

关键词：胃癌 5-氟尿嘧啶壳聚糖抗癌效果

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Effects of glycyrrhetinic acid on collagen metabolism of hepatic stellate cells at different stages of liver fibrosis in rats

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World Journal of Gastroenterology 2001年第1期7卷 115-119页

作者： ji yao wang Qi Sheng Zhang ji Sheng Guo Mei Yu Hu Department of Gastroenterology, Zhongshan Hospital, Medical Center, Fu Dan University Shanghai Medical University), Shanghai 200032, China Department of Internal Medicine Zhongshan Hospital Medical Center Fu Dan University Shanghai 200032 China.

INTRODUCTIONLiver fibrosis is a dynamic course leading tocirrhosis from a various chronic liver diseases. Thepathological basis of fibrosis is the disturbance ofproduction and degradation of the extracellularmatrix (E... 详细信息

关键词： liver cirrhosis glycyrrhetinic acid procollagen clostridium histolyticum collagenase extracellular matrix gene expression immunohistochemistry

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Auphen and dibutyryl cAMP suppress growth of hepatocellular carcinoma by regulating expression of aquaporins 3 and 9 in vivo

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World Journal of Gastroenterology 2016年第12期22卷 3341-3354页

作者： Rui Peng Guang-Xi Zhao jing Li Yu Zhang Xi-Zhong Shen ji-yao wang jian-Yong Sun Department of Gastroenterology Zhongshan Hospital Fudan University

AIM: To investigate whether the regulation of aquaporin 3(AQP3) and AQP9 induced by Auphen and dibutyryl c AMP(dbc AMP) inhibits hepatic tumorigenesis. METHODS: Expression of AQP3 and AQP9 was detected by Western blot, immunohistochemistry(IHC), and RT-PCR in HCC samples and paired non-cancerous liver tissue samples from 30 hepatocellular carcinoma(HCC) patients. A xenograft tumor model was used in vivo. Nine nude mice were divided into control, Auphen-treated, and dbc AMP-treated groups(n = 3 for each group). AQP3 and AQP9 protein expression after induction of xenograft tumors was detected by IHC and m RNA by RT-PCR analysis. The terminal deoxynucleotidyl transferase-mediated d UTP nick end labeling assay and histological evaluation were used to detect apoptosis of tumor cells, and the concentration of serum α-fetoprotein(AFP) was measured using RTPCR and an ELISA kit.RESULTS: The volumes and weights of tumors decreased significantly in the Auphen- and dbc AMP-treated mice compared with the control mice(P < 0.01). The levels of AQP3 were significantly lower in the Auphen treatment group, and levels of AQP9 were significantly higher in thedbc AMP treatment mice than in the control mice(P < 0.01). The reduction of AQP3 by Auphen and increase of AQP9 by dbc AMP in nude mice suppressed tumor growth of HCC, which resulted in reduced AFP levels in serum and tissues, and apoptosis of tumor cells in the Auphen- and dbc AMP-treated mice, when compared with control mice(P < 0.01). Compared with para-carcinoma tissues, AQP3 expression increased in tumor tissues whereas the expression of AQP9 decreased. By correlating clinicopathological and expression levels, we demonstrated that the expression of AQP3 and AQP9 was correlated with clinical progression of HCC and disease outcomes. CONCLUSION: AQP3 increases in HCC while AQP9 decreases. Regulation of AQP3 and AQP9 expression by Auphen and dbc AMP inhibits the development and growth of HCC.

关键词： Hepatocellular carcinoma Nude mice Auphen Dibutyryl c AMP Aquaporin 3 Xenograft tumor model Aquapori

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Investigating effect of coke porosity on blast furnace performance based on multi-physical fields

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Journal of Iron and Steel Research(International) 2024年第2期31卷 357-367页

作者： ji Chen Zhao Lei Zhe yao Li wang Qiang Ling Ping Cui Anhui Province Key Laboratory of Coal Clean Conversion and High Valued Utilization School of Chemistry and Chemical EngineeringAnhui University of TechnologyMa’anshan243002AnhuiChina Shanxi Academy of Printing Technology Research Institute Co. Ltd.Xi’an710077ShaanxiChina

Reducing coke use is an effective measure to reduce carbon emission and energy consumption in the blast furnace(BF)ironmaking.Essentially,BF is a high-temperature moving bed reactor,where complex physical transformations coupled with complicated reactions occur.This makes it challenging to investigate the factors determining BF performance with the conventional method.A multi-physical field coupling mathematical model of BF was thus developed to describe its mass and heat transfer as well as its intrinsic reactions.Then,the proposed model was validated with the production data.Under coupling conditions,influences of dominating reactions on BF performance(temperature distribution,gas distribution,iron formation reaction,and direct reduction degree)were revealed.The results indicated that coke combustion,indirect reduction,and direct reduction of iron ore mainly took place nearby the shaft tuyere,cohesive zone,and dripping zone,respectively.Besides,the rate of coke solution loss reaction was increased with the rising coke porosity in the cohesive zone.Considering the effect of coke porosity on the efficiency and stability of BF,the coke porosity of 0.42 was regarded as a reasonable value.

关键词： Blast furnace Mathematical model Multi-physical field Simulation Coke porosity

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Defects in the grain interiors of 3 mol%yttria-stabilized tetragonal zirconia polycrystal ceramics with 0.25 wt%alumina

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Journal of Advanced Ceramics 2024年第4期13卷 469-477页

作者： Yan Xiong Lian Luo yao Cheng Zhi Liu Qi Liu Weimin wang Wei ji School of Materials&Chemical Engineering Hubei Province Key Laboratory of Green Materials for Light IndustryHubei University of TechnologyWuhan 430068China Fujian Institute of Research on the Structure of Mater Chinese Academy of SciencesFuzhou 350002China jiangxi Provincial Engineering Research Center for Multifunction Zirconia Materials Jiangxi Size Materials Co.Ltd.Jiujiang 332500China State Key Laboratory of Advance Technology for Materials Synthesis and Processing Wuhan University of TechnologyWuhan 430070China

The presence of high-density defects is rarely observed in bulk 3 mol%yttria-stabilized tetragonal zirconia polycrystal(3Y-TZP)ceramics obtained through conventional pressureless sintering.In the present work,fine-grained dense 147 nm 3Y-TZP ceramics were prepared by pressureless sintering of commercial 0.25 wt%alumina-doped zirconia powders at 1300℃.A novel discovery was reported in which large amounts of defects were present in the grain interiors of the sample.The phenomenon was further examined using three types of powder samples,and the reasons for defect formation were investigated by microstructural characterization using high-resolution transmission electron microscopy(HRTEM)analysis and Rietveld refinement.The results confirmed the essential dependence of the defect formation on the alumina addition.The authors attributed the defect formation to the significant difference in ionic radii of the solvent and solute during the dissolution of alumina into the zirconia lattice.The sintering kinetics were proposed to be enhanced by the presence of substantial defects,which consequently favored the low-temperature sintering of the alumina-doped zirconia ceramics.

关键词： 3 mol%yttria-stabilized tetragonal zirconia polycrystal(3Y-TzP)ceramics alumina addition microstructure defect

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A robust viscoelastic surfactant tolerating 20% HCl up to 150℃ for oil well stimulation

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Petroleum Science 2024年第3期21卷 1865-1874页

作者： ji wang Ning Qi Hong-yao Yin Yu-Jun Feng Polymer Research Institute State Key Laboratory of Polymer Materials Engineering Sichuan University Tianfu Yongxing Laboratory New Theory and Technology of CO School of Petroleum Engineering China University of Petroleum (East China)

Viscoelastic surfactants（VES） are often used as viscous diverters in acidizing stimulation to prolong the acid consumption time and maximize zonal coverage of the acid for improving well productivity.However, the ceiling temperature of commercial VES cannot exceed 120℃ in practical use because of the poor thermal stability and fragile molecular structure, hindering their implementation in hightemperature oil reservoirs, i.e., ≥150℃. Here we synthesized a novel C22-tailed diamine, N-erucaminopropyl-N,N-dimethylamine（EDPA）, and examined comparatively its rheological behavior, assemblies morphology and molecular stability in 20 wt% HCl with a commercial VES, erucyl dimethyl amidopropyl betaine（EDAB）. The feasibility of EDPA for acidizing stimulation was assessed by acid etching of carbonate rock with its HCl solution at 150℃. Rheological results showed that the 2.5 wt% EDPA—20 wt%HCl solution maintains stable viscosity of 90 m Pa s at 150℃ for 60 min, while that of 2.0 wt% EDAB HCl solution is just 1 m Pa s under identical conditions.1H NMR spectra and cryo-TEM observations revealed that the chemical structure and self-assembled architectures of EDPA remained intact in such context,but the EDAB suffered from degradation due to the hydrolysis of the amide group, accounting for the poor heat-resistance and acid-tolerance. The reaction rate of 2.5 wt% EDPA HCl solution with carbonate rock was one order of magnitude lower than that of 20 wt% HCl solution at 150℃, underpinning the potential of EDPA to be used in the high-temperature reservoirs acidizing. This work improved the thermal tolerance of VES in highly concentrated HCl solution, paving a feasible way for the acidization of high-temperature reservoir environments（～150℃）.

关键词： Viscoelastic surfactants Thermal stability Self-diverting acid Wormlike micelles Acidizing stimulation

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