1.Vitro study on gene transfection efficiency of hyaluronic acid modified core-shell liponanoparticles in human retinal pigment epithelium cells.
Ya-Nan ZHAO ; Li GAN ; Jing WANG ; Xi CHEN ; Zheng JIA ; Yong GAN ; Jian-Ping LIU
Acta Pharmaceutica Sinica 2014;49(5):711-717
The aim of this study is to prepare hyaluronic acid (HA) modified core-shell liponanoparticles (pHA-LCS-NPs) as gene delivery system and investigate its gene transfection efficiency in human retinal pigment epithelium (ARPE-19) cells in vitro. The pHA-LCS-NPs was prepared by firstly hydrating dry lipid film with CS-NPs suspension to get LCS-NPs, then modifying the lipid bilayer with HA by amidation reaction between HA and dioleoyl phosphatidylethanolamine (DOPE). Its morphology, particle size and zeta potential were investigated. XTT assay was used to evaluate the cell safety of different vectors in vitro. The gene transfection efficiency of pHA-LCS-NPs modified with different contents of HA was investigated in ARPE-19 cells with green fluorescent protein (pEGFP) as the reporter gene. The results showed that the obtained pHA-LCS-NPs exhibited a clear core-shell structure with the average particles size of (214.9 +/- 7.2) nm and zeta potential of (-35 +/- 3.7) mV. The 24 h cumulative release of gene from pHA-LCS-NPs was less than 30%. After 48 h incubation, gene transfection efficiency of pHA-LCS-NPs/pEGFP was 1.81 times and 3.75 times higher than that of CS-NPs/pEGFP and naked pEGFP, respectively. Also no obvious cytotoxicity was observed on pHA-LCS-NPs. It suggested that the pHA-LCS-NPs might be promising non-viral gene delivery systems with high efficiency and low cytotoxicity.
Cell Survival
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Gene Transfer Techniques
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Genes, Reporter
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Genetic Vectors
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Green Fluorescent Proteins
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metabolism
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Humans
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Hyaluronic Acid
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chemistry
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pharmacology
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Lipids
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Nanoparticles
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Particle Size
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Phosphatidylethanolamines
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chemistry
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pharmacology
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Retinal Pigment Epithelium
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drug effects
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Transfection
2.Vitro study on gene transfection efficiency of hyaluronic acid modified core-shell liponanoparticles in human retinal pigment epithelium cells.
Yanan ZHAO ; Li GAN ; Jing WANG ; Xi CHEN ; Zheng JIA ; Yong GAN ; Jianping LIU
Acta Pharmaceutica Sinica 2014;49(5):711-7
The aim of this study is to prepare hyaluronic acid (HA) modified core-shell liponanoparticles (pHA-LCS-NPs) as gene delivery system and investigate its gene transfection efficiency in human retinal pigment epithelium (ARPE-19) cells in vitro. The pHA-LCS-NPs was prepared by firstly hydrating dry lipid film with CS-NPs suspension to get LCS-NPs, then modifying the lipid bilayer with HA by amidation reaction between HA and dioleoyl phosphatidylethanolamine (DOPE). Its morphology, particle size and zeta potential were investigated. XTT assay was used to evaluate the cell safety of different vectors in vitro. The gene transfection efficiency of pHA-LCS-NPs modified with different contents of HA was investigated in ARPE-19 cells with green fluorescent protein (pEGFP) as the reporter gene. The results showed that the obtained pHA-LCS-NPs exhibited a clear core-shell structure with the average particles size of (214.9 +/- 7.2) nm and zeta potential of (-35 +/- 3.7) mV. The 24 h cumulative release of gene from pHA-LCS-NPs was less than 30%. After 48 h incubation, gene transfection efficiency of pHA-LCS-NPs/pEGFP was 1.81 times and 3.75 times higher than that of CS-NPs/pEGFP and naked pEGFP, respectively. Also no obvious cytotoxicity was observed on pHA-LCS-NPs. It suggested that the pHA-LCS-NPs might be promising non-viral gene delivery systems with high efficiency and low cytotoxicity.
3.Case-control study on shoulder pain caused by hook palte for the treatment of acromioclavicular joint dislocation.
Ying-guo YANG ; Xiao-bing CAI ; Xiao-min WANG ; Yong-gan ZHU ; He-yong PAN
China Journal of Orthopaedics and Traumatology 2015;28(6):491-495
OBJECTIVETo explore causes of shoulder pain and propose prevention measures in treating acromioclavicular joint dislocation.
METHODSFrom January 2005 to January 2013, 86 patients with acromioclavicular joint dislocation (Tossy III) were treated with hook plate fixation, and were divided into two groups. Bsaed on recovery of shoulder function mostly, the patients who suffered from rest pain, motion pain were named as shoulder pain group, while the patients without pain were named as painless group. In shoulder pain group, there were 21 cases including 15 males and and 6 females ranging the age from 22 to 62 years old with an average of (40.6±11.2) years old. There were 8 cases were on the left side and 13 cases were on the right side. In painless group, there were 65 cases including 36 males and and 29 females ranging the age from 19 to 65 years old with an average of (40.0±11.3) years old. There were 33 cases were on the left side and 32 cases were on the right side. The time from injury to operation ranged from 3 h to 8 d with an average of 34.6 h. Shoulder function of all patients were normal before injuried. Postoperative pain, activity of daily living (ADL), range of motion, deltoid muscle strength were compared. Anteflexion,rear protraction, abduction and upthrow of shoulder joint were also compared. Postoperative complications between two groups were observed and compared.
RESULTSAll patients were followed up from 12 to 48 months with an average of 18.5 months. Constant-Murley score were used to evaluate clinical efficacy at the least following up, and 13 cases got an excellent results, 5 moderate, 2 good and 1 poor in shoulder pain group ; while 61 cases were obtained excellent results, 3 moderate and 1 good in painless group. There were significantly differences between two groups in Constant-Murley score and activity of shoulder joint (P<0.05). In shoulder pain group, 3 cases were disconnected, 1 case occurred stress fracture, 9 cases were subacromial impingement syndrome, 5 cases occurred subluxation, 1 case occurred plate breakage and 11 cases were acromioclavicular arthritis.
CONCLUSIONChosing individual clavicular hook plate, fulfilling anatomic reset, paying attention to the repair of articular capsule ligament, and reducing hook and bone antagonism between stress is the key point of preventing and decreasing postoperative shoulder pain.
Acromioclavicular Joint ; injuries ; physiopathology ; surgery ; Adult ; Bone Plates ; adverse effects ; Case-Control Studies ; Female ; Fracture Fixation, Internal ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Range of Motion, Articular ; Shoulder Dislocation ; complications ; physiopathology ; surgery ; Shoulder Pain ; etiology ; Treatment Outcome ; Young Adult
4.Primary osteosarcoma of kidney: report of a case.
Yong WU ; Xiaoqun YANG ; Hualei GAN ; Chaofu WANG
Chinese Journal of Pathology 2015;44(6):412-413
5.Preparation of Electrophoretic Microcolumn Packed with Quartz Microncrystal and Its Application to Electrophoretic Separation
Lian LI ; Youzhao HE ; Wuer GAN ; Xiaokui WANG ; Haiyang XIE ; Yong GAO
Chinese Journal of Analytical Chemistry 2009;37(7):1053-1056
The preparation of electrophoretic microcolumn and its application to the electrophoretic separation of amino acids with a 2-mm I.D. fused-silica microcolumn packed with uniform quartz microncrystal prepared by hydrothermal synthesis are reported. With 1.5 mmol/L disodium phosphate buffer solution (pH 11.5) containing 30% (V/V) methanol, the tryptophan, phenylalanine and tyrosine were separated by the microcolumn electrophoresis and detected by an UV spectrophotometer without derivatization. The limits of detection were 0.038, 0.21 and 0.20 mol/L, respectively. The separation efficiency of tryptophan was 4.4×104 plates/m. The sample capacity of the electrophoretic microcolumn achieved 35 μL. The precisions of the microcolumn electrophoresis were satisfactory. The thermal effects of the electrophoretic microcolumn that without packing, packed with 360 μm quartz sand and with 9 μm length quartz microncrystal were discussed, respectively. It was found that the electrophoretic microcolumn packed with quartz microncrystal was able to inhibit Joule heat, increase sample capacity and enhance detection sensitivity. The microcolumn electrophoresis is one of the high-performance separation techniques for an in-situ, real-time and portable electrokinetic flow analysis system.
6.Influences of silk fibroin on the compressive strength and injectability of calcium phosphate cement
Xiaoqing CHEN ; Huilin YANG ; Minfeng GAN ; Ruijuan XIE ; Liang CHEN ; Xuesong ZHU ; Genlin WANG ; Yong GU
Chinese Journal of Tissue Engineering Research 2008;12(45):8985-8988
BACKGROUND: High molecular materials have been proved to enhance the mechanical properties of calcium phosphate bone cement, as well as attenuate the injectability of composite materials. It thereby influences the clinical application of composite materials.OBJECTIVE: To observe the compressive strength and injectability of silk fihroin compound calcium phosphate bone cement, to evaluate the effect of silk fibroin on calcium phosphate, and to investigate the feasibility of applying silk fibroin as an injectable hone substitute to repair hone defects.DESIGN, TIME AND SETTING: Controlled study was performed in the central laboratory of Analysis and Testing Center, Soochow University from September to December in 2007.MATERIALS: Calcium phosphate cement was purchased from Shanghai Rebone Biomaterials Co., Ltd; silk fibroin was offered by Institute of Material & Engineering, Soochow University.METHODS: Six groups were set with different mass fractions (0.5%, 1%, 1.5%, 2%, 2.5%, 3%) of silk fibroin, which were mixed with bone cement at a certain liquid/solid ratio of 0.4 mL/g to prepare the calcium phosphate composite. The calcium phosphate cement without silk fibroin was served as control group.MAIN OUTCOME MEASURES: The compressive strength and injectability were determined. The characteristic microstructure was observed using scanning electron microscope.RESULTS: The compressive strength increased firstly and then decreased with the addition of silk fibroin. The compressive strength of the experimental groups was remarkably higher than the control group when the silk fibroin content was 1%-2.5% (P<0.05). The injectability of the paste diminished with the addition of silk fibroin, which was statistically different when the silk fibroin content was 1.5%-3% (P<0.05). Scanning electron microscope result revealed that the silk fihroin penetrated throughout calcium phosphate crystals, which were tightly connected.CONCLUSION: Silk fibroin can improve the compressive strength of silk fibroin/calcium phosphate cement composites without significant influence of manipulation, and can widen the application field of calcium phosphate bone substitute.
7.Evaluation of left ventricular torsion by two-dimensional ultrasound speckle tracking imaging in heart transplant patients
Shufen GAN ; Jianghua CHEN ; Yiqing WANG ; Zhian LI ; Yong JIANG ; Chongxian LIAO ; Lichun XU
Chinese Journal of Ultrasonography 2010;19(7):561-564
Objective To evaluate the value of left ventricular (LV) torsion for assessing left ventricular function in adult heart transplant patients using two-dimensional speckle tracking imaging.Methods Basal and apical LV short-axis view and apical LV long-axis view of two-demensional images were acquired in 30 heart transplant patients and 17 healthy volunteers. Using two-dimensional strain software,LV basal and apical rotation versus time profiles were obtained at their short-axis level respectively. LV torsion was defined as apical rotation relative to the base, so the LV torsion versus time profiles could be drawn. LV basal rotation,apical rotation,global torsion and their time to peak were respectively measured,and the rate of LV untwisting was calculated. Statistical analysis was used to find the difference between the two groups and to investigate the relationship between LV torsion or the rate of LV untwisting and echocardiographic parameters. Results LV apical rotation and LV global torsion in heart transplant group were significantly lower than those in normal group. LV global torsion inversely correlated with LV end systolic volume,positively with LV ejection fraction. There was no significant difference between the two groups in the rate of LV untwisting. The rate of LV untwisting didn't correlate with echocardiographic parameters. Conclusions LV torsion can be measured exactly by two-dimensional speckle tracking imaging, it can be used to quantify LV global systolic function in heart transplant patients.
8.Discussion on cultivation and methodology of four-drug combination-induced differentiation in mouse preadipocytes 3T3-L1 cells
Huizhi SUN ; Derun TIAN ; Jie MENG ; Nan ZHAO ; Jie HAN ; Chunchun GAN ; Yong WANG
Tianjin Medical Journal 2016;44(8):993-995
Objective To optimize and establish the methodology for culturing and inducing differentiation of mouse preadipocytes 3T3-L1. Methods The mouse cells 3T3-L1 were incubated in DMEM medium contained with 10%FBS, during which the incubation medium was refreshed every 2 to 3 days. Two methods were used to introduce differentiation, including three-drug combination group and four-drug combination group. The protocol of mediumⅠin three-drug combination group including insulin 10 mg/L, IBMX 0.5 mmol/L and DEX 1.0μmol/L. The protocol of mediumⅠin four-drug combination group including indometacin 0.1 mmol/L based on those of three-drug combination group. Both of them were incubated for 2 days and continuous for 2 times. And medium Ⅱ included insulin 10 mg/L for 2-day culturing and continuous for 2 times. Oil red O staining was used to observe the morphological changes of two groups of cells before and after treatment under inverted microscope. Results Mouse preadipocytes 3T3-L1 appeared in good conditions and grew in a paving stone fashion. These cells covered homogeneously the bottom of incubators, the culture medium refreshed every 2 days. The results of four-drug combination group were better than those of three-drug combination group. After three-drug combination induced differentiation, there was no significant change in cell morphology. Comparing with three-drug combination induced differentiation, four-drug combination was successfully achieved in over 90% of the cell inducing, which were round-shaped, with jacinth ester droplets by oil-red O staining. Conclusion We have optimized the method for culturing and inducing differentiation of mouse preadipocytes 3T3-L1 by adding indometacin on the basis of the three-drug combination induced differentiation.
9.Treatment of atrophic rhinitis by transplantation of pediculated bone-suberiosteal muscle flap
Yong-Gan WANG ; Qian-Mei SHI ; Yan-Hong WANG ; Chun-Jiu HU ; Zhong-Ming LIN ; Tao GUO ; Rong-Sheng NI ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(10):-
Objective To explore a better method for treatment atrophic rhinitis.Methods 56 patients with atrophic rhinitis(96 lateral)were treated by nasal submucou pediculated bone-suberiosteal muscle flap extracted from anterior wall of sinus maxillaries.Results All patients were followed 2 to 10 years,total effective rate was 100 %, with 49 cases(87.5 %)showing prominent effect.Conclusion The grafted flap cannot be assimilated,felled off and necrosis,because the flap has rich blood supply.This methods has obvious short-term effective and stable long-term effective.No complications were found.
10.Clinical evaluation of high viscosity bone cement in percutaneous vertebroplasty
Qian CHEN ; Dashou WANG ; Qi PAN ; Fengjun CAI ; Liang ZHANG ; Yong GAN ; Binwu DUAN ; Qianming WU ; Fei MENG
Chongqing Medicine 2014;(12):1457-1459
Objective To investigate the effect of high viscosity bone cement in percutaneous vertebroplasty to treat osteoporotic vertebral compression fracture(OVCF) and veterbral metastasis tumor(VMT) .Methods 102 cases of OVCF(OVCF group) and 16 cases of VMT(VMT group) in our hospital from March 2010 to December 2012 were performed percutaneous vertebroplasty under local anesthesia ,through the vertebral pedicle approach by conducting puncture at bilateral vertebral pedicle into the vertebra , 3-6 mL high viscosity bone cement was injected to each vertebra .The pain intensity was detected by the McGill pain questionnaire score before operation ,on postoperative 3 d ,1 month and 6 -18 months last follow-up .The preoperative and postoperative MPQ scores were statistically analyzed .Results Postoperative patients′s back pain was significantly alleviated or disappeared .Compared with before operation ,the MPQ scores on postoperative 3 d ,in 1 month and 6 -18 months last follow-up in the OVCF group and the VMT group were significantly decreased(P<0 .05) .The postoperative CT scanning revealed that the cement leakage rates in the OVCF group and the VM T group were 11 .76% and 6 .25% respectively .But no complications of bone cement toxicity ,pulmo-nary embolism ,infection and nerve damage occurred .Conclusion High viscosity bone cement injection has the advantages of strongly controllable injection due to high viscosity ,low bone cement leakage and high surgical safety ,which is suitable for the treatment of OVCF and VM T .