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In Vitro Functional Study of Rice Adenosine 5'-Phosphosulfate Kinase

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Rice science 2016年第3期23卷 152-159页

作者： Wang de-zhen Chen Guo-guo Lu Lu-jia Jiang Zhao-jun Rao Yu-chun Sun Mei-hao College of Chemistry and Life Sciences Zhejiang Normal University Jinhua 321004 China

Sulfate can be activated by ATP sulfurylase and adenosine 5'-phosphosulfate kinase(APSK) in vivo. Recent studies suggested that APSK in Arabidopsis thaliana regulated the partition between APS reduction and phosphorylation and its activity can be modulated by cellular redox status. In order to study regulation of APSK in rice(Os APSK), Os APSK1 gene was cloned and its activity was analyzed. Os APSK1 C36 and C69 were found to be the conserved counterparts of C86 and C119, which involved in disulfide formation in At APSK. C36A/C69 A Os APSK1 double mutation was made by site directed mutagenesis. Os APSK1 and its mutant were prokaryotically over-expressed and purified, and then assayed for APS phosphorylation activity. Os APSK1 activity was depressed by oxidized glutathione, while the activity of its mutant was not. Further studies in the case that oxidative stress will fluctuate in vivo 3'-phosphoadenosine-5'-phosphosulfate content, and all APSK isoenzymes have similar regulation patterns are necessary to be performed.

关键词： rice sulfate assimilation adenosine 5'-phosphosulfate kinase cysteine redox environment

GNOM-LIKE 2, Encoding an Adenosine Diphosphate-Ribosylation Factor-Guanine Nucleotide Exchange Factor Protein Homologous to GNOM and GNL1, is Essential for Pollen Germination in Arabidopsis

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Journal of Integrative Plant Biology 2009年第8期51卷 762-773页

作者： Dong-Jie Jia Xi Cao Wei Wang Xiao-Yun Tan Xue-Qin Zhang Li-Qun Chen de Ye State Key Laboratory of Plant Physiology and Biochemistry College of Biological Sciences China Agricultural University Beijing 100193 China National Center for Plant Gene Research Beijing 100101 China

In flowering plants, male gametes are delivered to female gametophytes by pollen tubes. Although it is important for sexual plant reproduction, little is known about the genetic mechanism that controls pollen germination and pollen tube growth. Here we report the identification and characterization of two novel mutants, gnom-like 2-1 （gnl2-1） and gn12-2 in Arabidopsis thaliana, in which the pollen grains failed to germinate in vitro and in vivo. GNL2 encodes a protein homologous to the adenosine diphosphate-ribosylation factor-guanine nucleotide exchange factors, GNOM and GNL1 that are involved in endosomal recycling and endoplasmic reticulum-Golgi vesicular trafficking. It was prolifically expressed in pollen grains and pollen tubes. The results of the present study suggest that GNL2 plays an important role in pollen germination.

关键词： adenosine diphosphate-ribosylation factor-guanine nucleotide exchange factor Arabidopsis gametophyte GNL2 male gametophytedefective pollen.

Value of dynamic 31p magnetic resonance spectroscopy technique in in vivo assessment of the skeletal muscle mitochondrial function in type 2 diabetes

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Chinese Medical Journal 2012年第2期125卷 281-286页

作者： WU Fei-yun TU Hui-juan QIN Bin CHEN Ting XU Hua-feng QI Jing WANG de-hang department of Radiology the First Affiliated Hospital of Nanjing Medical University Nanjing Jiangsu 210029 China

Background Phosphorous magnetic resonance spectroscopy （31p-MRS） has been successfully applied to study intracellular membrane compounds and high-energy phosphate *** study aimed to evaluate the capability of dynamic 31p-MRS for assessing energy metabolism and mitochondrial function in skeletal muscle from type 2 diabetic *** Dynamic 31p-MRS was performed on 22 patients with type 2 diabetes and 26 healthy *** were acquired from quadriceps muscle while subjects were in a state of rest,at exercise and during *** peak areas of inorganic phosphate （Pi）,phosphocreatine （PCr）,and adenosine triphosphate （ATP） were *** concentration of adenosine diphosphate （ADP） and the intracellular pH value were calculated from the biochemistry reaction *** time constant and recovery rates of Pi,PCr,and ADP were analyzed using exponential curve *** As compared to healthy controls,type 2 diabetes patients had significantly lower skeletal muscle concentrations of Pi,PCr and β-ATP,and higher levels of ADP and Pi/*** exercise,diabetics experienced a significant Pi peak increase and PCr peak decrease,and once the exercise was completed both Pi and PCr peaks returned to resting ***,the mean recovery rates of Pi and PCr in diabetes patients were （10.74±1.26） mmol/s and （4.74±2.36） mmol/s,respectively,which was significantly higher than in *** Non-invasive quantitative 31P-MRS is able to detect energy metabolism inefficiency and mitochondrial function impairment in skeletal muscle of type 2 diabetics.

关键词： type 2 diabetes phosphorus magnetic resonance spectroscopy skeletal muscle mitochondrial function

Amelioration of mitochondrial dysfunction in heart failure through S-sulfhydration of Ca2Vcalmodulin-dependent protein kinase II

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中国药理学与毒理学杂志 2017年第10期31卷 1025-1026页

作者： Dan WU Qing-xun HU de-qiu ZHU Yi-zhun ZHU department of Pharmacy Tongji Hospital Tongji University School of Medicine Shanghai 200065China School of Pharmacy Macao University of Science and Technology Macao China Mitochondria and Metabolism Center Department of Anesthesiology & Pain Medicine University of Washington Seattle WA 98109 USA

OBJECTIVE To determine the functional role of hydrogen sulfide (H2S) in protecting against mitochondrial dysfunction in heart failure through the inhibition of Ca2 +/calmodulin-dependent protein kinaseⅡ (CaMKⅡ) using wild type and CSE knockout mouse models. METHODS Continuous subcutaneous injection isoprenaline (7.5 mg·kg-1·d-1), once a day for 4 weeks to induce heart failure in Male C57BL/6 (6-8 weeks old) mice and CSE-/- mice. 150 μmol·L-1 H2O2 was used to induce oxidative stress in H9c2 cells. Echocardiograph was used to detect cardiac parameters. H&E stain and Masson stain was to observation histopathological changes. Western blot was used to detect protein expression and activity. The siRNA was used to silence protein expression. HPLC was used to detect H2S level. Biotin assay was used to detect the level of S- sulfhydration protein. RESULTS Treatment with S-propyl-L-cysteine (SPRC) or sodium hydrosulfide (NaHS), modulators of blood H2S levels, attenuated the development of heart failure in animals, reduced lipid peroxidation, and preserved mitochondrial function. The inhibition CaMKⅡ phosphorylation by SPRC and NaHS as demonstrated using both in vivo and in vitro models corresponded with the cardioprotective effects of these compounds. Interestingly, CaMKⅡ activity was found to be elevated in CSE-/- mice as compared to wild type animals and the phosphorylation status of CaMKⅡ appeared to relate to the severity of heart failure. Importantly, in wild type mice SPRC was found to promote S-sulfhydration of CaMKII leading to reduced activity of this protein however, in CSE-/- mice S-sulfhydration was abolished following SPRC treatment. CONCLUSION A novel mechanism depicting a role of S-sulfhydration in the regulation of CaMKⅡ is presented. SPRC mediated S-sulfhydration of CaMKII was found to inhibit CaMKⅡ activity and to preserve cardiovascular homeostasis.

关键词： hydrogen sulfide mitochondria heart failure Ca2 +/calmodulin- dependent protein kinaseⅡ S-sulfhydration

In vivo Evaluation of Titanium Implants Bioactivated by a Modified Kokubo’s Treatment

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Journal of Biosciences and Medicines 2014年第2期2卷 22-29页

作者： Jailson de Jesus Enori Gemelli Fábio Nery Nelson H. A. Camargo Materials Science and Engineering Department of Mechanical Engineering University of Santa Catarina State Campus Universitário Joinville Brazil

In this study a modified Kokubo’s treatment (MKT) was used to bioactivate titanium implants. Titanium surfaces were alumina-blasted and etched in HCl/H2SO4 solution followed by treatment in autoclave at 120°C for 1, 3, 5 hour periods with chemical solutions containing 1, 2 and 3 M of NaOH. The materials were characterized by surface angle measurements, roughness, field emission scanning electron microscopy (FE-SEM), X-ray photoelectron spectroscopy (XPS), and in vivo tests. Statistical analysis by polynomial regression showed that solutions within 2.2 and 2.8 M NaOH lead to surface contact angles close to zero degree after 1 h of treatment. XPS analysis carried out on a sample treated 1 h in 2.2 M NaOH indicated the presence of a titanate film. This solution was selected to treat titanium implants for in vivo tests. These tests were performed on sheep in three types of separating surfaces: machined group (standard), SLA group (A) and MKT group (B) divided into two time intervals of 3 and 6 weeks. The samples in group B displayed better results compared to group A only after 6 weeks in terms of counter torque and bone implant contact (BIC).

关键词： Titanium Thermochemical Treatment Wettability In vivo Test

Novel hybrid biocomposites for tendon grafts:The addition of silk to polydioxanone and poly(lactide-co-caprolactone)enhances material properties,in vitro and in vivo biocompatibility

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Bioactive Materials 2023年第7期25卷 291-306页

作者： Behzad Shiroud Heidari Emma Muiños Lopez Emma Harrington Rui Ruan Peilin Chen Seyed Mohammad Davachi Benjamin Allardyce Rangam Rajkhowa Rodney Dilley Froilán Granero-Moltó Elena M.de-Juan-Pardo Minghao Zheng Barry Doyle Harry Perkins Institute of Medical Research QEII Medical CentreNedlands and the UWA Centre for Medical ResearchThe University of Western AustraliaPerthAustralia School of Engineering The University of Western AustraliaPerthAustralia Australian Research Council Centre for Personalised Therapeutics Technologies Australia Cell Therapy Area Centro de Investigaci´on M´edica AplicadaIDISNAUniversidad de NavarraPamplonaSpain Centre for Orthopaedic Research Faculty of Health and Medical SciencesThe University of Western AustraliaPerthAustralia department of Biology and Chemistry Texas A&M International UniversityLaredoTXUSA Institute for Frontier Materials Deakin UniversityGeelongVIC3216Australia Perron Institute for Neurological and Translational Science NedlandsWestern AustraliaAustralia

Biopolymers play a critical role as scaffolds used in tendon and ligament(TL)*** advanced biopolymer materials have been proposed with optimised mechanical properties,biocompatibility,degradation,and processability,it is still challenging to find the right balance between these ***,we aim to develop novel hybrid biocomposites based on poly(p-dioxanone)(PDO),poly(lactide-co-caprolactone)(LCL)and silk to produce high-performance grafts suitable for TL tissue *** containing 1-15%of silk were studied through a range of characterisation *** then explored biocompatibility through in vitro and in vivo studies using a mouse *** found that adding up to 5%silk increases the tensile properties,degradation rate and miscibility between PDO and LCL phases without agglomeration of silk inside the ***,addition of silk increases surface roughness and *** vitro experiments show that the silk improved attachment of tendon-derived stem cells and proliferation over 72 h,while in vivo studies indicate that the silk can reduce the expression of pro-inflammatory cytokines after six weeks of ***,we selected a promising biocomposite and created a prototype TL graft based on extruded *** found that the tensile properties of both individual fibres and braided grafts could be suitable for anterior cruciate ligament(ACL)repair applications.

关键词： Biocomposite Tendon graft Silk Fibre extrusion Biodegradable scaffolds Anterior cruciate ligament

Combined use of chemotherapeutics and oncolytic adenovirus in treatment of AFP-expressing hepatocellular carcinoma

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Hepatobiliary & Pancreatic Diseases International 2009年第3期8卷 282-287页

作者： Mao, Chen-Yu Hua, Han-Ju Chen, Ping Yu, de-Chao Cao, Jiang Teng, Li-Song Zhejiang Univ Sch Med Dept Surg Oncol Affiliated Hosp 1 Hangzhou 310003 Peoples R China Zhejiang Univ Sch Med Sir Run Run Shaw Inst Clin Med Key Lab Biotherapy Zhejiang Prov Hangzhou 310016 Peoples R China

BACKGROUND: Hepatocellular carcinoma (HCC) which is always refractory to most chemotherapeutic agents may result in poor survival of patients with advanced HCC. Oncolytic adenovirus is a new form for cancer gene therapy via its ability to replicate and kill tumor cells in a tumor-specific manner. In order to eradicate tumors effectively, the combination of chemotherapeutic agents and oncolytic adenovirus has been considered. This study aimed to systematically analyze the possibility of synergistic cytotoxicity of oncolytic adenoviruses in combination with chemotherapeutic agents. METHODS: Several types of human HCC cell lines were used to determine the specificity and cytotoxicity of oncolytic adenovirus Ad5-HC and Ad5-AFP (IRES) by measuring cell viability in vitro and antitumor efficiency in vivo. The cytotoxicity of Ad5-HC and Ad5-AFP (IRES) combined with chemotherapeutic agents were also assessed by the methyl thiazolyl tetrazolium assay. RESULTS: Both Ad5-HC and Ad5-AFP (IRES) were significantly cytotoxic to HCC cells with great specificity in vitro and in vivo. The combination of oncolytic adenovirus with 5-FU, doxorubicin, and paclitaxel was synergistically effective for the killing of HCC cells. CONCLUSIONS: These data suggest that oncolytic adenovirus sensitize tumors to chemotherapy and the combination therapy of chemotherapeutic agents and oncolytic adenovirus has an enhanced antitumor effect on HCC cells. (Hepatobiliary Pancreat Dis Int 2009; 8: 282-287)

关键词： hepatocellular carcinoma drug resistance oncolytic adenovirus chemotherapy

Chronic stress sensitizes rats to pancreatitis induced by cerulein:Role of TNF-α

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World Journal of Gastroenterology 2010年第44期16卷 5565-5581页

作者： Marcelo G Binker Andres A Binker-Cosen Daniel Richards Herbert Y Gaisano Rodica H de Cosen Laura I Cosen-Binker CBRHC Research Health Center department of Medicine and Physiology University of Toronto

AIM:To investigate chronic stress as a susceptibility factor for developing pancreatitis,as well as tumor necrosis factor-α (TNF-α) as a putative ***:Rat pancreatic acini were used to analyze the influence of TNF-α on submaximal (50 pmol/L) cholecystokinin (CCK) *** restraint (4 h every day for 21 d) was used to evaluate the effects of submaximal (0.2 μg/kg per hour) cerulein stimulation on chronically stressed ***:In vitro exposure of pancreatic acini toTNF-α disorganized the actin *** was further increased by TNF-α/CCK treatment,which additionally reduced amylase secretion,and increased trypsin and nuclear factor-κB activities in a protein-kinase-C δ and ε-dependent ***-α/CCK also enhanced caspases' activity and lactate dehydrogenase release,induced ATP loss,and augmented the ADP/ATP *** vivo,rats under chronic restraint exhibited elevated serum and pancreatic TNF-α ***,pancreatic,and lung inflammatory parameters,as well as caspases' activity in pancreatic and lung tissue,were substantially enhanced in stressed/cerulein-treated rats,which also experienced tissues' ATP loss and greater ADP/ATP *** examination revealed that stressed/cerulein-treated animals developed abundant pancreatic and lung edema,hemorrhage and leukocyte infiltrate,and pancreatic *** severity was greatly decreased by treating animals with an anti-TNF-αantibody,which diminished all inflammatory parameters,histopathological scores,and apoptotic/necrotic markers in stressed/cerulein-treated ***:In rats,chronic stress increases susceptibility for developing pancreatitis,which involves TNF-α sensitization of pancreatic acinar cells to undergo injury by physiological cerulein stimulation.

关键词： Pancreatitis Stress Tumor necrosis factor-α

In vivo Preliminary Study on Bone Neoformation Behavior of Three Types of Calcium Phosphate Bioceramics

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Journal of Biosciences and Medicines 2014年第2期2卷 36-42页

作者： Nelson H. A. Camargo Enori Gemelli Aury N. de Moraes Bruna D. da Costa Nilson Oleskovicz Ademar L. Dallabrida Doughlas Regalin Marcos Paulo Antunes de Lima department Mechanical Engineering Santa Catarina State University Joinville Brazil department of Veterinary Medicine Santa Catarina State University Lagres Brazil

Calcium phosphate microporous bioceramics and biphasic compositions of hydroxyapatite and β-calcium phosphate, in the form of microporous granular biomaterials, are research topics and present themselves as potential orthopedic and biomedical applications in rebuilding and repairing maxillofacial bones and tooth structure. This is associated with the characteristics of microstructure, biocompatibility, bioactivity and bone conductivity properties which these materials offer when applied in vivo or in a simulation environment. This study aimed to assess the behavior of bone neoformation of three types of calcium phosphate biomaterials in in vivo tests with sheep within 60 and 90 days, with the help of a scanning electron microscope. The biomaterials used were provided by the Group of Biomaterials at the Santa Catarina State University. The in vivo tests were carried out by generating, on sheep, tibial bone defects, three of which were filled with biomaterial (one different biomaterial for each bone defect generated), whilst the fourth received a bone fragment obtained during the generation of the defect in question, to serve as a control group. The scanning electron microscopy (SEM) technique was used for carrying out the preliminary characterization studies so as to observe new bone formation and osseointegration. The X-ray diffractometry (XRD) served as a support for the characterization of crystalline phases. The results obtained are encouraging and show that the biomaterials presented good performance in the process of bone formation, biomaterial osseointegration by a new tissue and bone mineralization.

关键词： Bioceramics Microstructure Bone Neoformation Characterization