1.Effects of physical and chemical properties of polymeric nerve conduit on peripheral neuranagenesis.
Gao CHEN ; Jian-min ZHANG ; Hua HU ; Min LI ; Kang-jie ZHU
Journal of Zhejiang University. Medical sciences 2004;33(4):306-310
OBJECTIVETo evaluate the effects of degradation time and permeability of polymeric conduits on nerve regeneration.
METHODSAfter establishment of rat models with over 10 mm gap of sciatic nerve in right hind legs,four kinds of poly (ester, carbonate) nerve conduits was used to bridge the gaps and one group without conduit in gaps was used as control. The nerve regeneration and conduit degradation were examined both macroscopically and microscopically. The contraction of the muscle controlled by regenerated nerve was measured electrophysiologically at 4, 12 and 20 weeks after the operation.
RESULTBiodegradation time of nerve conduits in vivo was as fellows: 12 weeks in group A,4 weeks in group B and group C,and 20 weeks in group D,respectively. The histological quality of regenerative sciatic neurofibra in group A was the best among all groups (Mean rank 53.17, 38.83, 26. 60, 49.17 and 20.23,P<0.005), but the inflammatory reaction in group A was only less than that in group D and more than that in the other groups (Mean rank 45.87, 36.27, 34. 83, 51.63 and 21.4,P=0.001). The responsive rates of tibialis anterior muscle for electric stimulation in group A, B, C and D were 93.33%, 60%,20% and 73. 33%, respectively (P<0.005).
CONCLUSIONAbsorbable conduits with relatively good permeability and appropriate middle degrade time improve nerve regeneration and renovate function.
Action Potentials ; Animals ; Male ; Nerve Regeneration ; Peripheral Nerves ; physiology ; Rats ; Rats, Sprague-Dawley
2.Development of a system for quick screening of efficient HBx-siRNA.
Qin NI ; Ke-zhou LIU ; Zheng-gang YANG ; Hang-ping YAO ; De-hua AI ; Min-wei LI ; Zhi CHEN
Journal of Zhejiang University. Medical sciences 2004;33(4):300-310
OBJECTIVETo develop a system for quick screening of efficient siRNA targeted HBx mRNA.
METHODSUsing recombination DNA technique, the fusion expression plasmid of HBx and EGFP was constructed, and siRNA expression cassettes (SECs) containing U6+1, H1 or tRNA(Val )promoter were prepared via one-step overlapping extension PCR. By co-transfection with recombinant plasmid and SECs into AD293 cell, the inhibition effects on the transient expression of HBx-EGFP fusion protein were analyzed by FACS and semi-quantitated RT-PCR analysis.
RESULT(1)HBx-EGFP fusion protein expression plasmid pHBx-EGFP was constructed successfully, which expressed green fluorescence in cell mainly located at plasma or the periphery of nucleus in granules. (2) Co-transfection with recombinant plasmid and SECs into AD293 cells resulted in inhibition of HBx-EGFP expression. SEC-siHBx388 showed significant inhibition effect on HBx-EGFP expression compared with SEC-siHBx271, indicating that siHBx388 is effective siRNA site and could be screened out with our screening system. In addition,the results of that U6+1-, tRNA(Val) and H1-siHBx388 reduced HBx-EGFP expression by 21.7%, 12.9% and 12.4% of control respectively indicated that both tRNAVal and H1 promoter was high efficient in driving effect of siHBx388.
CONCLUSIONCombination of the HBx expression carrying reporter gene and PCR-based multi promoter SECs may develop a useful system to be applied in identification of optimal HBx- siRNA and its matching promoter.
Base Sequence ; Cells, Cultured ; Genetic Therapy ; Green Fluorescent Proteins ; Humans ; Luminescent Proteins ; genetics ; Molecular Sequence Data ; Plasmids ; RNA, Small Interfering ; analysis ; Recombinant Fusion Proteins ; biosynthesis ; Trans-Activators ; antagonists & inhibitors ; genetics ; Transfection
3.Expressions of bFGF, ET-1 and SCF in dermal papilla cells and the relation to their biological properties.
Zhong-fa LU ; Jin-jin WU ; Rong-qing LIU ; Bai-yu ZHONG ; Min ZHENG
Journal of Zhejiang University. Medical sciences 2004;33(4):296-299
OBJECTIVETo investigate the expression of bFGF, ET-1 and SCF in different passages of cultured dermal papilla cells (DPC), and their possible effect on biological behaviour of DPC.
METHODSThe expression of bFGF, ET-1 and SCF in different passages of cultured DPC was detected by immunocytochemistry and in situ hybridization.
RESULTThe expression of ET-1 and SCF in early passages of cultured DPC was stronger, but became negative in late passages (>6 passages). The stronger the expression of ET-1 and SCF in DPC, the higher ability of DPC to induce hair follicle regeneration.
CONCLUSIONThe expression strength of ET-1 and SCF is related to the ability of DPC inducing hair follicle regeneration.
Endothelin-1 ; analysis ; genetics ; Fibroblast Growth Factor 2 ; analysis ; genetics ; Hair Follicle ; chemistry ; cytology ; physiology ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Stem Cell Factor ; analysis ; genetics
4.Protective effects of minocycline against hair follicle damage induced by cytosine arabinoside in vitro.
Xian-jie WU ; Min ZHENG ; Zhong-fa LU
Journal of Zhejiang University. Medical sciences 2004;33(4):290-295
OBJECTIVETo investigate the protective effects of minocycline against hair follicle damage induced by cytosine arabinoside (Ara-c).
METHODSAn in vitro organ culture of mouse vibrissa follicles was used and different concentrations of Ara-c and minocycline were added in the culture media. The total growth length, growth speed and growth period of hair were observed with invert microscopy and the survival of hair bulb cells was measured by MTT method.
RESULTMinocycline (0.3 x 10(-6) approximately 10(-5) mol/L) improved hair follicle total growth length, growth speed and hair growth period and also improved survival of hair bulb cells in vitro organ culture, which were inhibited by Ara-c.
CONCLUSIONMinocycline can protect hair follicle directly from damage induced by Ara-c.
Animals ; Cytarabine ; toxicity ; Dose-Response Relationship, Drug ; Female ; Hair Follicle ; drug effects ; growth & development ; Male ; Mice ; Mice, Inbred C57BL ; Minocycline ; pharmacology
5.Hair follicle regeneration after implantation of hair follicle cells into subcutis of nude mice.
Zhong-fa LU ; Yue-lan CAO ; Min ZHENG
Journal of Zhejiang University. Medical sciences 2004;33(4):287-289
OBJECTIVETo observe the hair follicle regeneration after implantation of hair follicle cells into the subcutis of nude mice.
METHODSThe cultured hair papilla cells,dermal sheath cells and fibroblast of human scalp were mixed with the cells of hair follicle epithelium in different ratio, and then implanted into the subcutis of nude mice. The regeneration of hair follicle was observed.
RESULTThe hair follicle-like structure was formed in cluster where the cultured hair follicle epithelium cells were mixed with hair papilla cells. But no hair follicles were formed where the hair follicle epithelium was implanted with dermal sheath cells or fibroblasts.
CONCLUSIONThe hair follicle-like structure is generated in vivo when the mixed cells of early passages cultured hair papilla cells with hair follicle epithelium are implanted into the subcutis of nude mice.
Animals ; Cell Division ; Hair Follicle ; cytology ; physiology ; transplantation ; Mice ; Mice, Nude ; Regeneration
6.Skin-like structure generated from implantation of hair follicle bulb cells into collagen/chitosan porous scaffolds in vitro.
Xian-jie WU ; Zhong-fa LU ; Min ZHENG ; Yue-lan CHAO ; Sui-qing CAI ; Jian-guang ZHOU ; Lie MA ; Chang-you GAO
Journal of Zhejiang University. Medical sciences 2004;33(4):281-286
OBJECTIVETo observe the skin regeneration after hair follicle bulb cells were implanted into collagen/chitosan porous scaffolds in vitro.
METHODSThe cultured dorsal hair follicle bulb cells of 4d-old C57BL/6J mice were implanted into collagen/chitosan porous scaffolds in vitro. The skin regeneration was observed.
RESULTThe skin-like structure was formed on the collagen/chitosan porous scaffolds where were cultured the hair follicle bulb cells before 4th passages.
CONCLUSIONThe skin-like structure is generated in vitro when early passages of cultured hair bulb cells are implanted into collagen/chitosan porous scaffolds.
Animals ; Chitin ; analogs & derivatives ; Chitosan ; Collagen ; Hair Follicle ; cytology ; Mice ; Mice, Inbred C57BL ; Regeneration ; Skin ; cytology ; Tissue Engineering
8.Proliferation of neural stem cell and therapy of neuronal degenerative diseases.
Journal of Zhejiang University. Medical sciences 2004;33(3):272-276
Animals
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Cell Differentiation
;
drug effects
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Cell Division
;
drug effects
;
Ciliary Neurotrophic Factor
;
pharmacology
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Epidermal Growth Factor
;
pharmacology
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Genetic Engineering
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Humans
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Insulin-Like Growth Factor I
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pharmacology
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Neurodegenerative Diseases
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therapy
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Stem Cells
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cytology
9.CT evaluation of lymphoma in the abdominal lymph nodes.
Ri-sheng YU ; Shi-zheng ZHANG ; Jin-xiu WU ; Rong-fen LI
Journal of Zhejiang University. Medical sciences 2004;33(3):269-276
Adolescent
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Adult
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Aged
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Diagnosis, Differential
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Female
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Humans
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Lymph Nodes
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diagnostic imaging
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Lymphoma
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diagnostic imaging
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pathology
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therapy
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Male
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Middle Aged
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Tomography, X-Ray Computed