Forty-five healthy male dogs were wounded with high or middle-velocity steel ball on the lower part of the face.and the findings were as follows:(1)Craniocerebral injuries concomitant with maxillofacial gunshot wounds were charcterized by brain contusion in the entry side of the temporal lobe and extradural hemorrhage in the entry side of the middle cranial fossa,and their highest incidence and severity were found in those cases with mandible fracture due to high-velocity missiles.(2)The larger the amount of the absorbed energy from the bullet,the higher the incidence and severity of the craniocerebral injury.(3)The incidence and severity of the craniocerebral injury were positively correlated to the value of vibrational acceleration measured on the pareital bones,which suggests that vibrational acceleration plays an important role in the precipitation of craniocerebral injury secondary to maxillofacial gunshot wounds.

Objective To investigate the feasibility of labeling iris pigment epithelial(IPE)cells of rabbits with 5-(and 6-)carboxyfluorescein diacetate succinimidyl ester(CFSE). Methods Enzyme-assisted microdissection was used to isolate the cultured rabbit′s IPE cells.The third or forth subcultured IPE cells were incubated with 2.5,5,10,20,and 40 ?mol/L of CFSE for 1,5,and10min respectively.The fluorescence intensity was detected by flow cytometry,and the leakage of CFSE and its dyeing were observed by fluorescence antibody labeling. Results Incubation with 20 ?mol/L CFSE under 37℃for1minute was the most optimal condition for IPE cells labeling.The coloration of IPE cells stained by CFSE lasted 4 weeks.There was no leakage of dye from labeled rabbit IPE cells to non-labeled human IPE cells in mixed culture process. Conclusion With the advantages of high rate of dyeing,long time of tracing,safety and convenience,CFSE can be used as a new method to label the rabbit′s IPE cells.

To explore the roles of astrocytes in the epileptogenesis, astrocytes and neurons were isolated, purified and cultured in vitro from cerebral cortex of rats. The astrocytes were activated by ciliary neurotrophic factor (CNTF) and astrocytic conditioned medium (ACM) was collected to treat neurons for 4, 8 and 12 h. By using Western blot, the expression of calmodulin dependent protein kinase II (CaMK II), inducible nitric oxide synthase (iNOS) and adenylate cyclase (AC) was detected in neurons. The results showed that the expression of CaMK II, iNOS and AC was increased significantly in the neurons treated with ACM from 4 h to 12 h (P<0.05), and that of iNOS and AC peaked at 8 h and 12 h respectively. It was suggested that there might be some epileptogenic factors in the ACM and such signal pathways as NOS-NO-cGMP, Ca2+/CaM-CaMK II and AC-cAMP-PKA might take part in the signal transduction of epileptogenesis.

Quality is the lifeline of graduate education and the tutor team plays an important role in postgraduate training. To control the selection of tutors strictly, to implement the combination of openness and stability in tutor team, and to carry out academic exchanges actively are the keys to construct an excellent tutors staff and to ensure the quality of graduate student training.

BACKGROUND: There have been reports about the cochlea injury after high-velocity projectile wounding. The effect on ultrastructure of cochlea after blast was still unclear.OBJECTIVE: To investigate ultrastructural changes of cochlea and cochlear nerve after maxillofacial blast wound.DESIGN:A randomized controlled observational trail with dogs as subjects.SETTING: Otolaryngeal Department and Maxillofacial Surgery Laboratory of the Third Military Medical University of Chinese PLA.MATERIALS: The study was conducted from August 1995 through July 1997. The animal model in maxillofacial blast wound was established in Animal Center of Xinqiao Hospital of the Third Military Medical University of Chinese PLA. Specimens were treated in Maxillofacial Laboratory and observed in Electron Microscope Laboratory. Totally 15 dogs of either sex (weighting 9. 5 - 13.5 kg, mean 11.3 kg ) were randomly divided into three groups with 5 in each group. Two groups served as trauma groups 1 and 2 and the other group as control.METHODS: The maxillary and mandibular regions of 10 dogs in trauma groups were wounded by model 8 cardboard-shelled detonators to establish animal model of maxillofacial blast wound. At the 1st and 6th hour after trauma the wounds were examined and specimens of cochlea and cochlear nerve were dissected out for electron microscopic observation to study the ultrastructural changes. The specimens in the control group were treated the same way as those in the trauma groups except for blast injury.RESULTS: After wounded, the cochlea and cochlear nerve in the early period manifested cilia disorder, edema of the nerve and mitochondrial degeneration. At the 6th hour after trauma there were extensive degeneration in cochlea and cochlear nerve, cilia fallen off hair cells and dissolution of the structures in nerve sheath.CONCLUSION: The ultrastructural changes of cochlea and cochlear nerve are severed as a result of maxillofacial blast wound, but in early period the injury is reversible. So it is very important that early cure should be emphasized in treatment.

To localize and characterize the hypothetical protein CT358 in the chlamydial infected cells. CT358 gene from the Chlamydia trachomatis (C. trachomatis) serovar D genome was amplified and cloned into the pGEX and pDSRedCI vectors. The recombinant plasmid pGEX-CT358 was constructed and expressed as GST fusion proteins. The GST-CT358 fusion protein was used to immunize mice to raise the antibodies, which specifically recognized CT358 without eross-reacting with other unrelated proteins. The antibodies were then used to localize the endogenous CT358 protein and determine the expression pattern in Chlamydial infected cells using an indirect immunofluorescence assay (IFA). Meanwhile, pDSRedC 1-CT358 recombinant plasmid was transfected to HeLa cells to evaluate the effect of CT358 expression on the subsequent chlamydial infection. The hypothetical protein CT358 was identified in the inclusion membrane of C. trachomatis-infected cells for the first time,and it was detected as early as 12 h after C. trachomatis infection and remained in the inclusion membrane throughout the rest of the infection cycle. Cytosolic expression of CT358 via a transgene failed to affect the subsequent ehlamydial infection. These observations together have demonstrated that CT358 is a newly identified chlamydial inclusion membrane protein, giving the potentially importance for further understanding the mechanisms of chlamydial intracellular parasitism.

Objective To investigate the expression ofTNF- α mRNA and cytokeratin 20(CK20)mRNA in different tissue of colonic cancer patients, and the relations between the expression and the classify,invasion, as well as Dukes stage of colonic cancer. Methods RT-PCR method was used to detect the ex-pression of TNF-α mRNA and CK20 mRNA in 30 cases of colonic cancer, included cancer tissue,para-cancer tissue and normal tissue. Results The positive rate of TNF- α mRNA expressions in cancer tissue, para-cancer tissue and normal tissue were 70.0%, 43.3% and 20.0%, and the positive rate of CK20mRNA expressions were 63.3%, 33.3% and 16.7%, there were significant difference among the three tissues(P ＜ 0.01 ). But the expression of CK20 mRNA in para-cancer tissue had no significant difference compared with normal tissue (P＞ 0.05). The expression ofTNF- α mRNA was closely correlated with that of CK20mRNA.TNF- α mRNA and CK20 mRNA showed no significant difference in expressing of colonic cancer tissue (P ＞ 0.05 ), but TNF- α was closely correlated with Duke stage and depth of tumor invasion (P ＜ 0.05 ).Conclusion The expression of TNF- α mRNA is objective indicator associated with the invasion of the colonic cancer.

Objective:To investigate the immunogenicity of pORF5 plasmid protein,and further to screen and identify its im-munodominant domian.Methods: 10 different fragments of pORF5 gene including full length were amplified from the DNA of Chlamydia trachomatis serovar D by PCR and cloned into appropriate site of pGEX-6p vector to construct recombinant vectors after digested with BamHⅠand NotⅠrestriction endonucleases.After identification by PCR and sequencing,the recombinant plasmids were transformed into XL1 Blue E.coli to express the GST fusion proteins.ELISA and Western blot were carried out to identify the immunogenicity and immunoreaction of pORF5 plasmid protein.10 different fragments were reacted with sera from patients urogenitally infected with Chlamydia trachomatis, mouse polyclonal antibodies and mouse monoclonal antibodies of pORF5 plasmid protein with ELISA method.Results: pORF5 plasmid protein displayed strong immunogenicity and could induce a strong antibody response in human.The reactivity of human antibodies almost completely disappeared,when the native structure of pORF5 plasmid protein was de-stroyed.F6 that only lacked the N-terminal 66 amino acids was recognized by antibodies in ELISA as strongly as the whole pORF5 plasmid protein was.However,no other fragments were significantly recognized although there was a minimal reactivity of F2 and F3 with antibodies.Conclusion:pORF5 plasmid protein was an immunodominant antigen containing conformation-dependent epitope,and the C-terminal three quarters of pORF5 amino acid sequence was required for maintaining its immune dominance and conformation.The significance of the above findings lay a foundation for the further study on pORF5 protein function and vaccine development.

Objective To investigate the expression of eukaryotic initiation factor 4E(eIF4E) ,vascular endothelial growth factor (VEGF)‐A and VEGF‐C in gastric cancer tissues and their correlation with invasion and metastasis of gastric carcinoma .Methods The expressions of eIF4E ,VEGF‐A and VEGF‐C were detected in tissues of 58 gastric carcinomas and 25 normal gastric mucosa by using immunohistochemical method .Results The positive rate of eIF4E、VEGF‐A and VEGF‐C protein expression were 89 .7%(52/58) ,65 .5% (38/58) ,60 .3% (35/58) in gastric carcinoma ,which were higher than those in normal gastric mucosa tissues which were 4 .0% (1/25) ,12 .0% (3/25) ,8 .0% (2/25) respectively .Expressions of eIF4E ,VEGF‐A and VEGF‐C were significantly correlated with the depth of invasion and lymph node metastasis(P<0 .05) ,but not with the age ,sex of patients(P>0 .05) .Ex‐pressions of eIF4E and VEGF‐C were significantly correlated with tumor differentiation .The expression of eIF4E was being found positively correlated with VEGF‐A and VEGF‐C .Conclusion eIF4E may play certain roles in the oncogenesis and progression of gastric carcinoma .VEGF‐A and VEGF‐C may be helpful in lymph metastasis .Combined detection of eIF4E ,VEGF‐A and VEGF‐C may be helpful to assess the malignant degree and prognosis of gastric carcinoma .

Objective To construct the recombinant plasmid containing the major outer membrane protein(MOMP) gene of Chlamydia trachomatis and expres s MOMP protein in E.coli BL21. Methods The MOMP gene was amplified by polymera se chain reaction from the genome of Chlamydia trachomatis serovar D. The amplif ied fragment was directly inserted into pUCm-T vector and verified by DNA sequen cing. MOMP gene was then subcloned into the prokaryotic expression vector pET-22 b(+). The recombinant protein of MOMP was purified by Ni-NTA affinity chromatogr aphy and identified by SDS-PAGE and Western blot. Results The MOMP gene, which is about 1 200 bp, was successfully amplified and cloned. The DNA sequence of t he cloned MOMP gene was the same as that published by the GenBank. SDS-PAGE anal ysis showed that the relative molecular weight of this fusion protein was about 47 kDa which was consistent with the theoretically predicted value, and the spec ificity of this recombinant protein was confirmed by Western blot. Conclusions The MOMP gene of Chlamydia trachomatis was successfully cloned and expressed in the prokaryotic expression system, which may lay the foundation for the developm ent of Chlamydia trachomatis vaccine.