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Development of hybrid-type modified chitosan derivative nanoparticles for the intracellular delivery of midkine-siR NA in hepatocellular carcinoma cells

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Hepatobiliary & Pancreatic Diseases International 2015年第1期14卷 82-89页

作者： Jing Zhong Hui-Lian Huang Jing Li Fu-Chu Qian Li-Qin Li Ping-Ping Niu Li-Cheng Dai Huzhou Key Laboratory of Molecular Medicine Huzhou Central Hospital

BACKGROUND： Hepatocellular carcinoma（HCC） is one of the most common cancers worldwide. Most of the patients with HCC lose the surgical opportunity at the time of diagno sis. Some novel therapeutic modalities, like gene therapy, are promising for the treatment of HCC. However, the success of gene therapy depends on two aspects： efficient gene materials and gene delivery vectors. The present study was to develop new chitosan-based nanoparticles for a midkine-si RNA（anti HCC gene drug） ***： The novel gene delivery vector（MixN CH） was syn thesized by hybrid-type modification of chitosan with 2-chloro ethylamine hydrochloride and N, N-dimethyl-2-chloroethylamine hydrochloride. The chemical structure of Mix NCH was char acterized by FT-IR and 1HNMR. The cytotoxicity of Mix NCH was determined by MTS assay. The gene condensation ability and size, zeta potential and morphology of Mix NCH/MK si RNA nanoparticles were measured. The in vitro transfection and gene knockdown efficiency of midkine by Mix NCH/MK si RNA nanoparticles was detected by q RT-PCR and Western blotting. Gene knockdown effect at the molecule level on the proliferation of Hep G2 in vitro was determined by MTS ***： Mix NCH was successfully acquired by aminoalkyl ation modification of chitosan. The Mix NCH could condense MK-siR NA well above the weight ratio of 3. The average size of Mix NCH/MK-si RNA nanoparticles was 100-200 nm, and thesurface charge was about ＋5 m V. Morphologically, Mix NCH/MK-si RNA nanoparticles were in regular spherical shape-with no aggregation. Regarding to the in vitro transfection of nanoparticles, the MixN CH/MK-siR NA nanoparticles reduced MK m RNA level to 14.03%±4.03%, which were comparable to Biotrans（8.94%±3.77%）. Mix NCH/MK-si RNA effectively inhibited the proliferation of Hep G2 in vitro.-CONCLUSION： Mix NCH/MK-si RNA nanoparticles could be effective for the treatment of hepatocellular carcinoma.

关键词： chitosan gene vector nanoparticles midkine siRNA hepatocellular carcinoma

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midkine promotes perineural invasion in human pancreatic cancer

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World Journal of Gastroenterology 2014年第11期20卷 3018-3024页

作者： Jun Yao Wen-Yao Li Shuo-Guo Li Xiao-Shan Feng She-Gan Gao Department of Oncology First Affiliated HospitalHenan University of Science and Technology

AIM: To investigate midkine (MK) and syndecan-3 protein expression in pancreatic cancer by immunohistochemistry, and to analyze their correlation with clinicopathological features, perineural invasion, and prognosis.

关键词： midkine Syndecan-3 Pancreatic cancer Neurite growth Perineural invasion

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midkine ameliorates LPS-induced apoptosis of airway smooth muscle cells via the Notch2 pathway

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Asian Pacific Journal of Tropical Biomedicine 2022年第12期12卷 512-519页

作者： Qi-Feng Huang Bo Wang Yu-Qing Weng Tang Deng Li-Hua Li Jin Qian Qi Li Kai-Wen Lin Dong-Mei Sun Shuang-Qin Xu Hang-Fei Wang Xin-Xin Wu Yuan-Tian Sun Xiao-Ran Liu Emergency Department The First Affiliated Hospital of Hainan Medical UniversityHaikou 570102China Jiangdong Health Center Haikou 571126China Department of Pulmonary and Critical Care Medicines Zhuhai People’s Hospital(Zhuhai Hospital Affiliated with Jinan University)Zhuhai 519050China College of Pharmacy Hainan Medical UniversityHaikou 570100China Shandong Heze Mudan People’s Hospital Heze 274007China Emergency and Trauma College Hainan Medical UniversityHaikou 570100China College of Basic Medicine and Life Sciences Hainan Medical UniversityHaikou 570100China Key Laboratory of Emergency and Trauma of Ministry of Education Haikou 570100China

Objective:To evaluate the effect of midkine on lipopolysaccharide(LPS)-induced airway smooth muscle cells(ASMCs).Methods:LPS-stimulated acute lung injury model was used to analyze the effect of midkine on ASMCs in *** midkine and midkine siRNA were used to investigate the role of Notch2 signaling *** proliferation was assessed using Cell Counting Kit-8 ***,apoptosis was measured by flow cytometry and protein and mRNA expression of midkine and Notch2 was assessed by Western blotting and qPCR,*** analysis was also ***:LPS increased the mRNA and protein expression of midkine and *** silencing reduced LPS-induced midkine and Notch2 *** addition,midkine silencing further reduced the viability and increased apoptosis of ASMCs induced by LPS,which was attenuated by recombinant ***:The midkine/Notch2 signaling pathway plays a regulatory role in ASMC proliferation and apoptosis in airway inflammation.

关键词： Acute lung injury Airway smooth muscle cells midkine Notch2 Lipopolysaccharide

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midkine expression in 52 human meningiomas:A correlation analysis

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Neural Regeneration Research 2008年第10期3卷 1141-1144页

作者： Xinjun Li Xiangguo Xia Department of Neurosurgery People＇s Hospital of Deyang City Deyang 618000 Sichuan Province China Department of Neurosurgery the Affiliated Hospital of Luzhou Medical College Luzhou 646000 Sichuan Province China

BACKGROUND： Several studies have shown that midkine directly participates in tumor cell growth and invasion, as well as the regulation of angiogenesis. OBJECTIVE： To investigate midkine expression in meningioma tissue in relation to angiogenesis, invasion, peritumoral edema, and clinicopathology. DESIGN, TIME AND SETTING： The present clinical, case-controlled, neuropathological study was performed at the Laboratory of Molecular Organism, People＇s Hospital of Deyang City between May 2007 and April 2008. MATERIALS： Fifty-two meningioma tissues were classified by WHO tumor classification of the central nervous system, comprising 40 grade Ⅰ meningioma, five grade Ⅱ meningioma, and seven grade Ⅲ meningioma. Ten normal, human cerebral maters were selected from cerebral trauma patients. METHODS： midkine protein expression and mean microvessel density were detected using immunohistochemical techniques. Simultaneously, all data were statistically analyzed. MAIN OUTCOME MEASURES： midkine expression and microvessel density in meningiomas and normal cerebral maters. RESULTS： The positive midkine expression rate was 64% in the meningioma tissues. However, midkine expression was not detected in normal cerebral mater tissue. The mean microvessel density was 82.0 ± 22.7 in the meningiomas, and 25.8± 6.2 in the normal cerebral mater tissues. There was significant difference in midkine expression and mean microvessel density between meningioma tissues and human cerebral maters （P 〈 0.05）. midkine expression positively correlated with microvessel density （r = 0.756, P 〈 0.05）. midkine expression did not correlate to patient age, gender, or tumor size, location, and shape （P 〉 0.05）. However, it closely correlated with patient clinical condition, pathological grade, invasion, and peritumoral edema （r = 0.3785,0.741 2,0.6518, 0.614 2, P 〈 0.05）. CONCLUSION： midkine protein was overexpressed in meningiomas and correlated to tumor angiogenesis, invasion, pefitumoral edema, and clinicopath

关键词： CD34 immunohistoehemical meningioma microvessel density midkine

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*** mycelium inhibits epithelial–mesenchymal transition by inactivating the midkine pathway in pulmonary fibrosis

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Frontiers of Medicine 2021年第2期15卷 313-329页

作者： Li Lu Haiyan Zhu Hailin Wang Huaping Liang Yayi Hou Huan Dou The State Key Laboratory of Pharmaceutical Biotechnology Division of ImmunologyMedical SchoolNanjing UniversityNanjing 210093China Department of Rheumatology and Immunology Nanjing Drum Tower HospitalThe Affiliated Hospital of Nanjing University Medical SchoolNanjing 210008China State Key Laboratory of Trauma Burns and Combined InjuryResearch Institute of SurgeryDaping HospitalThe Army Medical UniversityChongqing 400042China Jiangsu Key Laboratory of Molecular Medicine Nanjing UniversityNanjing 210093China

The medical fungus Hirsutella sinensis has been used as a Chinese folk health supplement because of its immunomodulatory *** previous studies established the antifibrotic action of Hirsutella sinensis mycelium(HSM)in the *** epithelial–mesenchymal transition(EMT)is involved in the pathogenesis of idiopathic pulmonary *** present study investigates the role of HSM in mediating EMT during the development of pulmonary *** significantly inhibits bleomycin(BLM)-induced pulmonary fibrosis by blocking the *** addition,the expression levels of midkine are increased in the lungs of the BLM-induced *** analysis of the results indicates that the mRNA level of midkine correlated positively with *** markedly abrogates the transforming growth factorβ-induced EMT-like phenotype and behavior in *** activation of midkine related signaling pathway is ameliorated following HSM treatment,whereas this extract also caused an effective attenuation of the induction of EMT(caused by midkine overexpression)in *** further confirm that oral medication of HSM disrupted the midkine pathway in ***,findings suggest that the midkine pathway and the regulation of the EMT may be considered novel candidate therapeutic targets for the antifibrotic effects caused by HSM.

关键词： epithelial-mesenchymal transition H.sinensis mycelium midkine pulmonary fibrosis

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Large-scale proximity extension assay reveals CSF midkine and DOPA decarboxylase as supportive diagnostic biomarkers for Parkinson’s disease

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Translational Neurodegeneration 2023年第1期12卷 315-328页

作者： Wojciech Paslawski Shervin Khosousi Ellen Hertz Ioanna Markaki Adam Boxer Per Svenningsson Laboratory of Translational Neuropharmacology Department of Clinical NeuroscienceKarolinska InstitutetStockholmSweden Memory and Aging Center University of CaliforniaSan FranciscoSan FranciscoCAUSA Basic and Clinical Neuroscience Institute of PsychiatryPsychology and Neurosci-enceKing’s College LondonLondonUK Old Age Psychiatry Institute of PsychiatryPsychology and NeuroscienceKing’s College LondonLondonUK.

Background There is a need for biomarkers to support an accurate diagnosis of Parkinson’s disease(PD).Cerebrospinal fluid(CSF)has been a successful biofluid for finding neurodegenerative biomarkers,and modern highly sensitive multiplexing methods offer the possibility to perform discovery *** a large-scale multiplex proximity extension assay(PEA)approach,we aimed to discover novel diagnostic protein biomarkers allowing accurate discrimination of PD from both controls and atypical Parkinsonian disorders(APD).Methods CSF from patients with PD,corticobasal syndrome(CBS),progressive supranuclear palsy(PSP),multiple system atrophy and controls,were analysed with Olink PEA *** cohorts were used in this study,comprising 192,88 and 36 cases,*** samples were run on the Cardiovascular II,Oncology II and Metabolism PEA *** Our analysis revealed that 26 and 39 proteins were differentially expressed in the CSF of test and validation PD cohorts,respectively,compared to *** them,6 proteins were changed in both ***(MK)was increased in PD with the strongest effect size and results were validated with *** most increased protein in PD,DOPA decarboxylase(DDC),which catalyses the decarboxylation of DOPA(L-3,4-dihydroxyphenylalanine)to dopamine,was strongly correlated with dopaminergic ***,Kallikrein 10 was specifically changed in APD compared with both PD and controls,but unchanged between PD and *** inhibitory factor 1 was consistently downregulated in CBS and PSP patients in two independent *** Using the large-scale PEA approach,we have identified potential novel PD diagnostic biomarkers,most notably MK and DDC,in the CSF of PD patients.

关键词： Atypical Parkinsonian disorders Biomarker Cerebrospinal fluid Corticobasal syndrome DOPA decarboxylase midkine Multiple system atrophy Parkinson’s disease Progressive supranuclear palsy Proximity extension assay

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Antisense oligonucleotides targeting midkine induced apoptosis and increased chemosensitivity in hepatocellular carcinoma cells

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Acta Pharmacologica Sinica 2006年第12期27卷 1630-1636页

作者： Li-cheng DAI~(2,5)Xiang WANG~2 Xing YAO~3 Yong-liang LU~3 Jin-liang PING~4 Jian-fang HE~2 2 Huzhou Key Laboratory of Molecular Medicine,3 Department of General Surgery,4 Department of Pathology,Huzhou Central Hospital,Huzhou 313000,China Huzhou Key Laboratory of Molecular Medicine Huzhou Central Hospital Huzhou China Department of General Surgery Huzhou Central Hospital Huzhou China Department of Pathology Huzhou Central Hospital Huzhou China

Aim:Overexpression of midkine(MK)has been observed in many *** aim of this study is to screen for suitable antisense oligonucleotides(ASODN)targeting MK in hepatocellular carcinoma(HCC)cells and evaluate its ***:Ten ASODN targeting MK were designed and *** transfection with ASODN,cell proliferation was analyzed with MTS[3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,inner salt]*** addition,MK mRNA,protein levels,as well asapoptosis and caspase-3 activity were also examined in HepG2 *** prolif-eration was then analyzed after treatment with both ASODN and chemotherapeu-tic ***:In this experiment,the ASODN5 among the 10 ASODN showedhigher inhibitory activity against proliferation of hepatocellular carcinoma cells ina dose-dependent *** HepG2 cells,ASODN5 could significantly reducethe MK mRNA level and protein *** transfection with ASODN5 for48h,accompanied with a decline of survivin and Bcl-2 protein content,a remark-able increase of apoptosis and caspase-3 activity was observed in HepG2 ***,ASODN5 transfer can significantly increase chemosensitivity inHepG2 ***:Antisense oligonucleotides targeting MK shows thera-peutic effects on HCC;ASODN5 has the possibility to be developed as an effec-tive antitumor agent.

关键词： antisense oligonucleotide midkine apoptosis hepatocellular carcinoma chemosensitivity

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Novel functional proteins interact with midkine in hepatic cancer cells

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Hepatobiliary & Pancreatic Diseases International 2012年第3期11卷 272-277页

作者： Qiang Yan, Hui-Lian Huang, Xing Yao, Jing Li, Li-Qin Li, Jing Zhong, Li-Shan Min, Li-Cheng Dai and Shu-Sen Zheng Department of Hepatobiliary and Pancreatic Surgery, Huzhou Central Hospital (Yan Q and Yao X) Huzhou Key Laboratory of Molecular Medicine, Affiliated Central Hospital of Huzhou Teachers College (Huang HL, Li J, Li LQ, Zhong J, Min LS and Dai LC), Huzhou 313000, China Division of Hepatobiliary and Pancreatic Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China (Yan Q and Zheng SS) [a] Department of Hepatobiliary and Pancreatic Surgery Huzhou Central Hospital Huzhou 313000 China [b] Huzhou Key Laboratory of Molecular Medicine Affiliated Central Hospital of Huzhou Teachers College Huzhou 313000 China [c] Division of Hepatobiliary and Pancreatic Surgery First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China

BACKGROUND: midkine is a heparin-binding growth factor that promotes the proliferation, survival, migration and differentiation of various target cells. midkine plays an important role in tumorigenesis and tumor progression, and is overexpressed in many human malignant tumors. Patients with high tumor midkine expression frequently have a worse prognosis than those with low expression. The present study was designed to investigate the interaction network of midkine in hepatic cancer cells, and to elucidate its role in hepatocellular carcinoma. METHODS: DNA encoding full-length midkine was cloned into pDBLeu vector to serve as bait in yeast two-hybrid screening to identify interacting proteins. Candidate proteins were examined on SC-Leu-Trp-His+3-AT (20 mmol/L) plates and assayed for X-gal activity, then sequenced and classified according to the GenBank. Finally, identified proteins were expressed by the in vitro expression system pCMVTnT, and protein interactions were confirmed by co-immunoprecipitation. RESULTS: Using the yeast two-hybrid system, we found 6 proteins that interacted with midkine: NK-kappa-B inhibitor alpha (I-κ-B-α), Dvl-binding protein naked cuticle 2, granulin, latent active TGF-β binding protein 3, latent active TGF-β binding protein 4, and phospholipid scramblase 1. In vitro co-immunoprecipitation demonstrated that all identified proteins directly interacted with ***: The identification of midkine-interacting proteins in hepatic cancer cells indicates that midkine is a multifunctional factor that may participate in cell migration, differentiation, and proliferation, and is also associated with the multicellular response feedback during the development of hepatocellular carcinoma.

关键词： midkine yeast two-hybrid interaction hepatic cancer cell

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EXPRESSION OF midkine IN BENIGN, PREMALIGNANT AND MALIGNANT VULVAR TUMORS

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Chinese Medical Sciences Journal 2002年第3期17卷 148-152页

作者：武昕姚嘉斐李祁伟郑华川辛彦 Department of Gynecology The First Clinical College China Medical University Shenyang 110001 Department of Pediatric Orthopedics The Second Clinical College China Medical University Shenyang 110003 Cancer Institute The First Clinical College China Medical University Shenyang 110001

OBJECTIVE: To clarify the role of midkine (MK) in vulvar carcinogenesis though examination of its expression in vulvar lesions including vulvar condyloma acuminata (VCA), vulvar intraepithelial neoplasia (VIN) and vulvar squamous cell carcinomas (VSCC), and to analyze the relationship between MK expression and human papilloma virus (HPV) infection. METHODS: Thirty VSCC, 15 VIN and 10 VCA patients were studied by streptavidin-biotin-immunoperoxidase method. MK expression was compared with clinicopathologic features of vulvar tumors. RESULTS: MK was expressed in 26 of 30 VSCC (87%), 3 of 5 VIN III and all VCA samples, whereas no MK expression was detected in the VIN I-II samples or in normal epithelium. The difference of MK expression between VIN III and VSCC was statistically significant (P

关键词： midkine vulvar squamous cell carcinoma vulvar condyloma acuminataObjective. To clarify the role of midkine (MK) in vulvar carcinogenesis through examination of its expression in vulvar lesions including vulvar condyloma acuminata (VCA), vulva

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Serum midkine levels for the diagnosis and assessment of response to interventional therapy in patients with hepatocellular carcinoma

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Journal of Interventional Medicine 2021年第1期4卷 39-45页

作者： Lin Zheng Hailiang Li Jinhua Huang Jihoon Shin Suxia Luo Chenyang Guo Yan Zhao Fangkun Li Department of Minimally Invasive Intervention Henan Cancer HospitalThe Affiliated Cancer Hospital of Zhengzhou UniversityZhengZhou450008China Division of Medical Imaging and Interventional Radiology Sun Yat-sen University Cancer CenterGuangzhou510060China Department of Radiology and Research Institute of Radiology University of UlsanCollege of MedicineAsan Medical Center86Asanbyeongwon-gilSongpa-guSeoul 138-736Republic of Korea Department of Medical Gastroenterology Henan Cancer HospitalThe Affiliated Cancer Hospital of Zhengzhou UniversityZhengZhou450008China

Objective:To explore the clinical significance of serum midkine(MDK) levels for the diagnosis of hepatocellular carcinoma(HCC) and evaluate the efficacy of interventional ***:Eighty-four patients with HCC were enrolled in this retrospective *** received an interventional treatment.A follow-up was performed every 2 months,using magnetic resonance imaging,to determine whether the treatment should be *** alpha-fetoprotein(AFP) and MDK levels were measured at the first diagnosis and during the follow-ups,and the HCC detection rates based on the cutoff values of these two measurements were *** relationships between AFP and MDK and the clinical tumor characteristics and changes in APK and MDK before and after treatment were also compared using a rank sum test and χ2 test,*** prognostic significance of MDK for HCC was determined through regression analysis.A twosided P <0.05 was considered statistically ***:MDK expression was detected in 95.24% of the *** analysis revealed MDK expression in95.35%,95.12%,85.19%,86.67%,and 83.33% of the AFP-positive,AFP-negative,stage A Barcelona clinic liver cancer(BCLC-A),BCLC-A/AFP-positive,and BCLC-A/AFP-negative cases,*** expression after the interventional treatment(66.7%) was significantly lower than that before the treatment(95.2%).The mean posttreatment MDK level was 0.67 ng/mL in patients with a positive response to therapy as compared with 3.66 ng/mL in those with no positive *** patients were followed up for 18 months,and those positive for MDK expression before the intervention were more likely to relapse than patients without MDK *** analysis revealed the highest recurrence rate for patients who were positive for MDK expression before and after ***:Serum MDK may serve as a powerful complement to AFP in the diagnosis of *** measurement may improve the detection rate of BCLC-A and AFP-negativ

关键词： Hepatocellular carcinoma midkine Interventional treatment Diagnosis Efficacy evaluation

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