2.Transport of PLGA nanoparticles across Caco-2/HT29-MTX co-cultured cells.
Zhen WEN ; Gang LI ; Dong-Hai LIN ; Jun-Teng WANG ; Li-Fang QIN ; Gui-Ping GUO
Acta Pharmaceutica Sinica 2013;48(12):1829-1835
The present study is to establish Caco-2/HT29-MTX co-cultured cells and investigate the transport capability of PLGA nanoparticles with different surface chemical properties across Caco-2/HT29-MTX co-cultured cells. PLGA-NPs, mPEG-PLGA-NPs and chitosan coated PLGA-NPs were prepared by nanoprecipitation method using poly(lactic-co-glycolic acid) as carrier material with surface modified by methoxy poly(ethylene glycol) and chitosan. The particle size and zeta potential of nanoparticles were measured by dynamic light scattering. Coumarin 6 was used as a fluorescent marker in the transport of nanoparticles investigated by confocal laser scanning microscopy. The transport of furanodiene (FDE) loaded nanoparticles was quantitively determined by high performance liquid chromatography. Colchicine and nocodazole were used in the transport study to explore the involved endocytosis mechanisms of nanoparticles. Distribution of the tight junction proteins ZO-1 was also analyzed by immunofluorescence staining. The results showed that the nanoparticles dispersed uniformly. The zeta potential of PLGA-NPs was negative, the mPEG-PLGA-NPs was close to neutral and the CS-PLGA-NPs was positive. The entrapment efficiency of FDE in all nanoparticles was higher than 75%. The transport capability of mPEG-PLGA-NPs across Caco-2/HT29-MTX co-cultured cells was higher than that of PLGA-NPs and CS-PLGA-NPs. Colchicine and nocodazole could significantly decrease the transport amount of nanoparticles. mPEG-PLGA-NPs could obviously reduce the distribution of ZO-1 protein than PLGA-NPs and CS-PLGA-NPs. The transport mechanism of PLGA-NPs and mPEG-PLGA-NPs were indicated to be a combination of endocytosis and paracellular way, while CS-PLGA-NPs mainly relied on the endocytosis way. PEG coating could shield the surface charge and enhance the hydrophilicity of PLGA nanoparticles, which leads mPEG-PLGA-NPs to possess higher anti-adhesion activity. As a result, mPEG-PLGA-NPs could penetrate the mucus layer rapidly and transport across Caco-2/HT29-MTX co-cultured cells.
Biological Transport
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Caco-2 Cells
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Chitosan
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chemistry
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Coated Materials, Biocompatible
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chemistry
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Coculture Techniques
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Drug Carriers
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Furans
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administration & dosage
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chemistry
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metabolism
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HT29 Cells
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Heterocyclic Compounds, 2-Ring
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administration & dosage
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chemistry
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metabolism
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Humans
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Lactic Acid
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chemistry
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Nanoparticles
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Particle Size
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Polyethylene Glycols
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chemistry
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Polyglycolic Acid
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chemistry
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Zonula Occludens-1 Protein
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metabolism
3.Transgenic Tobacco Expressing the Crocus CSzCD Gene Showed the Production of Crocetin
Hai-Bo XIAO ; Lan WANG ; Chen YAN ; Ying XU ; Fang CHEN ; Lin TANG ;
China Biotechnology 2006;0(06):-
To investigate the feasibility to produce crocetin in tobacco plants.The coding region of zeaxanthin cleavage dioxygenase (CSzCD) gene from Crocus sativus L. was inserted into the downstream of the cauliflower mosaic virus (CaMV) 35S promoter of a binary vector pBI121, and integrated into the genome of Nicotiana tabacum L. Twenty-one transgenic lines were identified by genomic southern blotting analysis. Western blotting and HPLC analysis of the leaf extracts of transgenic tobacco showed that crocetin was produced in CSzCD gene-transformed plants, while no crocetin was found in the untransformed tobacco.
4.Study of Expression of Pneumococcal Surface Protein and Immunogenicity
Qian-Ying CAI ; Liang FANG ; Jin-Zhong HUANG ; Hai-Ying LIN ; Yang-Hao GUO ; Chun MENG ;
China Biotechnology 2006;0(04):-
The specific fragment of Pneumococcal surface protein A(PspA)and Pneumococcal Surface Adhesin A(PsaA)gene was amplified by PCR from Streptococcus pneumonia 5 and Streptococcus pneumonia 19.The amplified fragnent of PspA and PsaA gene was ligated into pET-27b(+)vector and transformed into BL 21 E.coli for expression and obtain the expressive production of PspA and PsaA.Induced by IPTG,the expression level was as high as 75 % of the total disolube protein.The result showed that the recombinant plasmid could express a specific 75 kDa and 37 kDa fusion protein in E.coli BL 21,which showed the good immunogenicity and a broadly cross reactivity with the other serotypes.
5.Evaluation of neutralizing antibodies in serum immunized with virus-like particle chimerized HCV series of neutralizing epitopes
Xiaoyan WANG ; Hai ZHANG ; Yingfeng LEI ; Fang LIN ; Ying CUI ; Bin LI ; Huizhong ZHANG ; Sanhua WEI
Chinese Journal of Immunology 2017;33(5):707-711,720
Objective:New Zealand rabbits were immunized with VLPs-MEpS,VLPs-E2S,and the levels of neutralizing antibodies in serum were determined.Methods: New Zealand rabbits were immunized with 10 μg VLPs-MEpS and VLPs-E2S,serum was collected at diffferent time with a two-weeks interval.The neutralizing antibodies were determined by ELISA.HCV(type 1b) had been prepared and mixed with serum from immunized rabbit before infected Huh7.5 cell.The protection of neutralizing antibodies in serum was assessed.Results: Neutralizing antibodies had been induced in rabbit after immunized with VLPs-MEpS and VLPs-E2S.VLPs-MEpS group had higher titer of antibodies than that of VLPs-E2S group(P<0.05),both group had higher titer of antibodies than that of control groups significantly(P<0.01).VLPs-MEpS group had higher neutralization than that of VLPs-E2S group(P<0.05),the highest neutralization rate was 61.49%.Both groups were higher than control group notably(P<0.01).Conclusion: Protective neutralizing antibodies have been induced in New Zealand rabbit after immunized with VLPs-MEpS and VLPs-E2S.It′s the basement for development of neutralizing antibodies vaccine.
6.Observation of Testis Tissue in Experimental Cryptorchidism after Orchiopexy in Various-Day-Rats
cui-ping, SONG ; hai-lei, WANG ; guo-lin, ZHANG ; fang, AI ; jin-lan, CHEN
Journal of Applied Clinical Pediatrics 2006;0(23):-
ObjectiveTo observe testis tissue changes of experimental cryptorchidism after orchiopexy in various-day-rats.MethodsSeventy-two Sprague-Dawley(SD) male rats were divided into 3 groups randomly and made artificial cryptorchidism:unilateral cryptorchidism group(n=24),bilateral cryptorchidism group(n=24)and sham operation group(n=24)at age 21-day-old.Intra-abdominal testicle was resetting in 2 weeks,at age 40 days and 60 days,the rats were sacrificed for detection tubular fertility index(TFI) and mean tubular diametar(MTD) with hematoxylin-eosine staining and germ cell apoptosis by terminal deoxynucleotidyl transferase mediated d-UTP nick end labeling(TUNEL) assay.ResultsThere were significant differences of MTD,TFI and apoptosis index(AI) between cryptorchidism and scrotal testes(P0.05).The AI of cryptorchidism testes of unilateral cryptorchidism group were significantly lower than that of bilateral cryptorchidism group in 40 days(P0.05).ConclusionsAI of artificial resetting testis is increased and contralateral descended testes in unila-teral cryptorchidism have various damage.It is to lighten that pathological damage of testes of cryptorchidism with prolongation of reset time.
7.Open reduction and mini-plate internal fixation for the treatment of fracture and dislocation of coccyx.
Hai-bo FANG ; Hong-hai XU ; Hai-lin FANG ; Yue-lin ZHANG ; Zong-zhi LIU ; Fan-xing WANG ; Qing-yang ZOU
China Journal of Orthopaedics and Traumatology 2013;26(7):549-552
OBJECTIVETo explore the therapeutic methods of fracture and dislocation of coccyx and evaluate its curative effects.
METHODSFrom May 2002 to March 2010,56 patients with fracture and dislocation of coccyx were divided into surgical treatment group and non-surgical treatment group. There were 7 males and 20 females in surgical treatment group with an average age of (48.1 +/- 0.6) years (ranged, 29 to 62 years), treated with open reduction and mini-plate internal fixation. There were 8 males and 21 females in non-surgical treatment group with an average age of (47.5 +/- 0.9) years (ranged, 19 to 54 years),treated with manipulative reduction. All patients were underwent X-ray examination and were finally diagnosed before treatment. Clinical symptoms and Visual Analogue Scales (VAS) of all patients were statistically analyzed before and after treatment.
RESULTSThere was no significant difference between two groups in gender, age, BMI index and VAS evaluation. All patients were followed up from 12 to 25 months with an average of 17.2 months. In surgical treatment group,there were 26 cases with I/a incision and 1 case with II/a incision; the excellent rate of clinical symptom was respectively 92.6% and 100% at leaving hospital and final follow-up; the improvement rate of VAS was 97.6% and was excellent result;internal fixtures were removed at the 1 to 2 years after treatment and no unwell symptoms occurred; VAS of all patients in the group was 0 point. In non-surgical treatment group,the excellent rate of clinical symptom was respectively 72.4% and 82.8% at leaving hospital and final follow-up; the improvement rate of VAS was 72.1% and was good result. There was significant difference in clinical results between two groups (P<0.05).
CONCLUSIONThe results indicated that fracture and dislocation of coccyx should be treated in time. For the treatment of patients with manipulative reduction failures, instability reduction by X-ray examination and serious rectal irritation, open reduction and mini-plate internal fixation can obtain satisfactory results.
Adult ; Bone Plates ; Coccyx ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Joint Dislocations ; surgery ; Male ; Manipulation, Spinal ; Middle Aged ; Spinal Fractures ; surgery
8.Chemical constituents of the roots of Macleaya microcarpa and activation efficacy of benzophenanthridine alkaloids for the transcription of xbp1 gene.
Yang LIU ; An-Jun DENG ; Lin MA ; Hai-Jing ZHANG ; Zhi-Hui ZHANG ; Lian-Qiu WU ; Zhu-Fang SHEN ; Wen-Jie WANG ; Hai-Lin QIN
Acta Pharmaceutica Sinica 2015;50(2):207-210
Ongoing study on the chemical constituents of the roots of Macleaya microcarpa led to the isolation of eight compounds of derivatives of triterpenes and organic acids in addition to some previously identified benzophenanthridines. The eight compounds were identified by spectroscopic methods as well as comparison with literature values as 1-oxo-2, 22 (30)-hopandien-29-oic acid (1), 3-oxo-12-oleanen-30-oic acid (2), 3α-hydroxy-12-oleanen-30-oic acid (3), 3β-hydroxy-12-oleanen-30-oic acid (4), ferulic acid (5), ferulic acid 4-O-β-D-glucoside (6), 3-O-feruloylquinic acid (7), and methyl 3-O-feruloylquinate (8). Of which, 1 is a new triterpenoid of hopanes and 2-8 are isolated from M microcarpa for the first time. In order to discover natural active compounds as potential agents of anti-ulcerative colitis (UC), an in vitro drug high-throughput screening model targeted x-box-binding protein 1 (xbp1) was employed to evaluate the activity of the major chemical constituents of M microcarpa. The result confirmed that two dihydrobenzophenanthridines, dihydrosanguinarine (9) and dihydrochelerythrine (10), showed a certain activity on activating the transcription of xbpl, a transcription factor (TF) associated with the occurrence, development, and potential treatment of UC, with their relative activating ratios being 1.76 and 1.77 times, respectively, as compared with control group.
Anti-Ulcer Agents
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chemistry
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Benzophenanthridines
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chemistry
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DNA-Binding Proteins
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genetics
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Isoquinolines
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chemistry
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Papaveraceae
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chemistry
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Plant Roots
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chemistry
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Regulatory Factor X Transcription Factors
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Transcription Factors
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genetics
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Transcription, Genetic
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Triterpenes
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chemistry
9.Study on Biological Characters of Sindbis Virus Yunnan Strain
Wen-li, HUANG ; Hai-lin, ZHANG ; Jing-lin, WANG ; Yu-zeng, ZHANG ; Zhu-qin, MI ; Hua-fang, SHI ; Deng-yun, ZI ; Guo-lin, ZHOU ; Guo-dong, LIAN
Virologica Sinica 2001;16(2):97-100
The biological characters of Sindbis virus strain of Yunnan(YN87448 strain) were studied by the test of the filtration, acid-resistant, ether-resistant, CPE, susceptibility of animal, HA, plague, determination of virus titres, and the cross-HI, cross-IFAT and PRNT as well.The results indicated that YN87448 strain belongs to Sindbis virus, Alphavirus, Togaviridae. YN87448 strain virus was plaque purified(PYN87448). The biological character of PYN87448 strain virus was studied too. PYN87448 strain virus will be used in the molecule biological test.
10.Optic radiation in normal adults: a study using magnetic resonance diffusion tensor imaging and diffusion tensor tractography.
Si-hai WAN ; Xue-lin ZHANG ; Xin-lan XIAO ; Xin SUN ; Hai-fang XING ; Shi-jun QIU
Journal of Southern Medical University 2008;28(3):396-398
OBJECTIVETo study the fractional anisotropy (FA) and the architecture of the optic radiation fiber tracts of normal adults with magnetic resonance (MR) diffusion tensor imaging (DTI).
METHODSDiffusion tensor images were obtained from 30 healthy volunteers without any cerebral abnormalities on conventional MRI. FA and the mean diffusivity (MD) of the optic radiation were measured in the directional encoded color (DEC) maps. The architecture of the optic radiation fiber tracts were displayed with the software of diffusion tensor fiber tracking.
RESULTSIn all subjects, the optic radiation could be readily identified in the DEC maps. The FA value was 0.509-/+0.029 in the left and 0.502-/+0.026 in the right, with the MD value of (0.763-/+0.050) x10(-3) and 0.748-/+0.052)x10(-3) mm2/s, respectively. No significant differences were found in the FA or MD value of the bilateral optic radiation (P>0.05). Diffusion tensor tractography (DTT) demonstrated that the 3 bundles of the optic radiation fibers were located in the lateral sagittal stratum, passing from the lateral geniculate body of the thalamus to the primary visual cortex. The dorsal and lateral bundles passed posteriorly to the superior bank of the calcarine cortex, while the ventral bundle passed anteriorly before making a sharp turn, known as the Meyer loop, and subsequently coursed posteriorly to terminate in the inferior margin of the calcarine cortex, which was consistent with the results of classic anatomical studies.
CONCLUSIONAs a novel method to study the relationship between visual function and optic pathway, DTI and DTT can show the FA and architecture of the optic radiation.
Adult ; Anisotropy ; Diffusion Magnetic Resonance Imaging ; methods ; Echo-Planar Imaging ; methods ; Female ; Geniculate Bodies ; anatomy & histology ; Humans ; Male ; Middle Aged ; Models, Anatomic ; Occipital Lobe ; anatomy & histology ; Optic Nerve ; anatomy & histology ; Visual Pathways ; anatomy & histology ; Young Adult