[Objective]Using a procedure of chemical agent to remove the cells and myethin in spinal cord of rat and to prepare the scaffold of extracellular matrix,so as to obtain an ideal natural spinal cord scaffold to bridge the nerve gap.[Method]Rat spinal cord was cut and treated using the method of freeze thawing and chemical extraction(3%sodiumdeoxycholate and 1KU/ml DNaseI,RNaseA).Histology was exploited to evaluate the degree of acellular and the structure of the spinal cord scaffold.[Result]In cross section,network of the extracellular matrix was presented in the scaffold.The cells,myethin and axons disappeared after the spinal cord was treated with sodium deoxycholate and DNaseI,RNaseA.Typical network of empty tubes were viewed in longitudinal sections.[Conclusion]An ideal spinal cord scaffold can be produced with the method designed in authors experiment.This scaffold has similar three dimensional structure with normal spinal cord,which can be used as a graft to bridge the nerve gap directly or as a scaffold to implant the seeding cells in spinal cord tissue engineering.The experiment indicates that cells and myethin can be removed and the three dimensional structure be reserved by chemical extraction with 3% sodium deoxycholate and 1KU/ml DNaseI,RNaseA.Chemical extraction is an ideal method to prepare tissue engineer scaffold of spinal cord.

[Objective] To investigate the immunogenicity of the acellular spinal cord scaffold and to provide theoretical basis for its further application in tissue engineering.[Method]Acellular spinal cord(freeze thawing +3%sodiumdeoxycholate + DNaseI 、RNaseA)and fresh spinal cord of rats were implanted into paravertebral muscles of rats.The tissue was obtained at 1、2、3 and 4w after the operation,then the inflammatory reaction was evaluated by HE stain and the immunogenicity of acelular scaffold was tested by immunohistochemical examination of the intensity of CD3+ 、CD4+ and CD8+ cells that infiltrated the allografts.[Result]The bistological examination indicated that acellular spinal cord scaffold was surrounded by a amount of neutrophilic cells and lymphocytes one week postoperatively,yet two weeks postoperatively,there was only small amount of lymphocytes infiltration.Fresh spinal cord allograft elicited an intense acute inflammatory infiltrate,and two weeks later,there still had a mount of lymphocytes infiltration.The intensity of CD3+、CD4+ and CD8+ T cells that infiltrated the allografts was greatly lower in acellular spinal cord than that in fresh spinal cord.The mild cell-mediated host-graft immune rejection in acellular spinal cord was observed.[Conclusion]The acellular spinal cord scaffold has mild inflammatory reaction and immune rejection,suggestting it is qualified for some biological properties and it may be a potential alternative scaffold of tissue engineering.

Objective To evaluate the in vitro biological safety of acellular spinal cord scaffold so as to provide theoretical basis for constructing the ideal tissue engineering scaffold of spinal cord.Methods A piece of thoracic spinal cord for 2 cm removed from SD rats was harvested and then was treated by freezing and thawing and chemical extraction with 3% sodium deoxyeholate and 1 KU/ml DNaseI and RNaseA. Gross observation and histological examination of the acellular spinal cord scaffold were carried out to learn the condition of the extracellular matrix scaffold. The biological safety of the acellular spinal cord scaffold was evaluated. Results In cross section, network of the extracellular matrix was presented in the scaffold. The cells, myelin and axons disappeared after the spinal cord was treated with sodium deoxycholate, DNaseI and RNaseA. Typical network of empty tubes were viewed in longitudinal sections. General toxic reaction, pyrogen test, hemolysis test and cytotoxicity test were conforming to the standard of materials. Conclusion As neotype tissue engineering material, the acellular spinal cord scaffold has satisfactory biological safety.

Objective To compare the effect of 2 decellularizing methods,sodium deoxycholate plus Triton X-100 or plus DNase and Rnase,in the preparation of acelluar allograft spinal cord scaffold in order to provide an ideal natural spinal cord scaffold to bridge the nerve gap.Methods Spinal cord was removed from health rats,and then decellularized by the method of freeze thawing(immersing in 3% sodium deoxycholate followed by the mixture of 1?103 U/ml DNase and RNase),or by chemical extraction(immersing in 1% Triton X-100 and then 1% sodium deoxycholate).HE staining,myelin staining and scanning electron microscopy(SEM) were employed to evaluate the spinal cord scaffold after the 2 methods of decellularization.Results Both cells and myelin were completely decellularized with the 2 methods.In cross section,network of the extracellular matrix was presented without axon,sheath and cells nucleus being seen in the scaffold.Typical network of empty tubes were viewed in longitudinal sections.Conclusion An ideal spinal cord scaffold can be produced with these 2 decellularizing methods in tissue engineering.The scaffold made by the 2 methods have similar three dimensional structure with normal spinal cord,so can be used as a graft to bridge the nerve gap directly or as a scaffold to implant the seeding cells in spinal cord tissue engineering.

Objective To investigate expression of glycogen synthase kinase(GSK)-3? and phosphorylation GSK-3? (p-GSK-3?) in the anterior temporal neocortex in patients with refractory epilepsy(RE). Methods Expression of GSK-3? and p-GSK-3? were detected by RT-PCR, FQ-PCR, Western Blot and immunohistochemistry in the anterior temporal cortices of 36 RE cases. 8 patients without RE had been used as the controls. Results Compared with control group,the expression of GSK-3? mRNA and protein were significant higher(allP

Marine traditional Chinese medicine (MTCM) is an important part of Chinese medicine (CM),there are some differences in understanding of the current literature and the extension of the connotation of marine medicine,which leads to the definition dispute of MTCM,hindering clinical application and further development of MTCM.In this study,we explored the concept of MTCM in literature,discussed the attributes of ocean marine CM,summed up the differences between the land CM and MTCM over variety characteristics,effect of drug composition characteristics and biological activity characteristics,and discussed the connotation and extension of MTCM from three aspects of theoretical basis and the effect and source of the drug,leading to the formation of the narrow and broad concept of MTCM.The five kinds of disputes in the definition of MTCM were discriminated according to the concept and connotation,which provided a theoretical basis for the definition and the research of MTCM.Moreover,we also defined the English translation and its abbreviation as Marine Traditional Chinese Medicine (MTCM).

Objective To prepare the acellular scaffold of spinal cord and analyze its component.Methods The acellular scaffold was prepared with the freeze thawing and chemical extraction,its structure was observed by HE and SEM,its component was analyzed by immunohistochemistry.Results The cells,myelin sheath and axon of nerve fibers in the rat spinal cord were eliminated,but three-dimensional supports of extracellular matrix were reserved.The analytical results showed the component of the acellular spinal cord contain laminin,fibronectin and type Ⅳ collagen—the necessary components to facilitate and induce the regeneration of the injured nerves and enhance the adhesion and proliferation of cells.Conclusion The acellular spinal cord has three dimensional structure and contains several proteins related to the regeneration of the injured nerves and promotion of the survival and proliferation of cells.

Objective To provide anatomical evidence for the repair of wounds of finger distal phalanx,espe-cially for the recovery of feeling. Methods 10 samples of fresh adult hand were dissected under microscope. The course,branches,distribution and external diameter of nerves and blood vessels in finger distal phalanx and morpho-logical relationship between nerves and vessels were measured. Results Proper palmar digital nerves mostly step over digital arteries at section starts of distal finger arterial arcades and go to finger pulps and latero-backs. Their thinks di-vide into 2 branches. Transverse diameters of interior and exterior branches are 0.8 ～ 1.2 mm and 0.9 ～ 1.4 mm re-spectively at liner semilunaris levels. Distributionsof left and right branches are reciprocal chiasmas. Conclusion Finger nerve mostly ramifies to finger pulp,finger tip and finger back at the level of phalangette bottom. Its branches are lower and thinner than concomitant arteries. The suitable anatomy region for anastomosis of nerves and blood ves-sels is the middle1/3 section from the distal interphalangeal joint to the nail during replanation of amputated finger pa-ratelum.

OBJECTIVE To investigate the vicissitudes of resistance to imipenem in Gram-negative bacilli isolated from clinical specimens.METHODS Gram-negative bacilli were isolated from various clinical specimens in our hosptial from Jan 2004 to Jan 2008.The identification and antimicrobiol susceptiblity test was determined by VITEK-AMS full automated microbiology analyzer.RESULTS A total of 6983 strains of Gram-negative bacilli were isolated,including 92 species and 2986 isolates of Enterobacteriaceae and 3961 isolates of non-fermenters,accounted for 42.8% and 56.7%,respectively.The resistant rate of Enterobacteriaceae to imipenem was lower than 1.0%.The resistant rates of Stenotrophomonas maltophilia,Burkholderia cepacia,Chryseobacterium meningosepticum,Pseudomonas aeruginosaand Acinetobacter baumannii to imipenem were 98.4%,97.0%,98.2%,70.1% and 47.5%,respectively.CONCLUSIONS Non-fermenters are predominant pathogens.Imipenem has high antimicrobial activity to Enterobacteriaceae in invro,but not to non-fermenters.The resistant rates of P.aeruginosa and A.baumannii to imipenem are increasing gradually.

OBJECTIVE To study the distribution and antimicrobial resistance of Acinetobacter baumannii infection in our hospital.METHODS Five hundred and sixty-nine strains of A.baumannii isolated from patients with infection from Jan 2001 to Dec 2005 were collected and antimicrobial susceptibility tests were performed.RESULTS Two hundred and seventy-five strains(48.3%) of A.baumannii were from intensive care unit(ICU).Four hundred and five strains(71.2%) of A.baumannii were examined from sputum.A.baumannii had various drug resistances to 12 antibiotics,which were monitored and proved tending to strengthen.The resistance rate in the ICU was distinctly higher than the others with significant difference(P