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.

Objective To evaluate the effect of dexmedetomidine on cognitive dysfunction after thoracic surgery in patients undergoing one-lung ventilation.Methods Sixty-two patients,aged 45-64 yr,of ASA physical status Ⅰ or Ⅱ,with body mass index ranged from 17.5 to 25.5 kg/m2,scheduled for elective thoracic surgery,were randomly allocated into 2 groups (n =31 each) using a random number table:dexmedetomidine group (Dex group) and control group (C group).Dexmedetomidine 0.5 μg/kg was infused for 10 min starting from the time point before induction of anesthesia,followed by continuous infusion at a rate of 0.5 μg · kg-1 · h-1 until 30 min before the end of surgery in Dex group.The equal volume of normal saline was administered instead in group C.Before induction and at 24,48 and 72 h after surgery,venous blood samples were collected for determination of levels of S-100 beta protein and neuronspecific enolase in serum by ELISA.Cognitive function was assessed by Mini-Mental State Examination at 72 h after surgery.Results The levels of S-100 beta protein and neuron-specific enolase in serum were significantly increased after surgery than before induction in the two groups.Compared to C group,the levels of S-100 beta protein and neuron-specific enolase in serum were significantly decreased after surgery,and the incidence of postoperative cognitive dysfunction was decreased in Dex group (26％ vs 6％).Conclusion Dexmedetomidine can effectively reduce the nerve damage during one-lung ventilation and significantly inhibit the development of postoperative cognitive dysfunction in patients undergoing thoracic surgery,indicating that dexmedetomidine is suitable for thoracic surgery.

Objective To construct DNA vaccine containing MOMP gene of Chlamydia trachomatis and to observe immune response in mice. Methods Mice of 4 - 6 weeks old were immunized with pcDNA3.1-MOMP or pcDNA3.1 intramuscularly at a dose of 100 μg. Booster immunizations were employed at 2-week interval for two times. Specific antibody in the sera of mice and the level of IFN-γ in murine spleen lymphocyte supernatant were detected by ELISA. The proliferation response of spleen cells was detected by MTT assay. Results Significant specific antibody titers were observed and the highest titer was 1: 1 024 in mice after three times immunization with pcDNA3.1-MOMP. The proli-feration response of spleen cells were significantly higher than that of mice injected with pc DNA3.1. IFN-γ reached(532.0 + 45.4)pg/mL in immunized mice. Conclusion Strong responses of humoral and cellular immunity can be evoked by DNA vaccine of pcDNA3.1-MOMP in mice.

Objective:To investigate the antitumor efficiency of the special cytotoxic T lymphocytes(CTLs) activated by dendritic cells(DCs) pulsed with K-ras (12-Val) antigen.Methods:DCs was generated from PBMC in the presence of granuloceyte/macrophage-colony stimulating factor(GM-CSF),interleukin-4(IL-4)in vitro.DCs were differently sensitized with K-ras mutant pancreatic cancer cell line,K-ras(12-Val) mutant peptide,K-ras(12-Val) mutant peptide with the surface of cationic nanoparticle.Cell surface markers on DCs was measured by flow cytometry.The activation of CTL induced by DCs was detected by ~3H- thymidine incorporation test.The killing effects of CTL to pancreatic cancer was detected by ~(125)I-UdR release test. Production of IL-12 and IFN-γ by DCs and PBMC was detected by ELISA.Results:Compared with DCs pulsed with K-ras(12-Val) mutant peptide and K-ras (12-Val) mutant peptide with the surface of cationic nanoparticle,DCs pulsed with whole tumor antigen could better induce CTLs killing activity(P＜0.05).The DCs with K-ras(12-Val) mutant peptide and K-ras mutant peptide with the surface of cationic nanoparticle could produce specific CTL killing activity aganist pancreatic cancer cell line Patu8988(K-ras+)(P＜0.05),but not SW1990(K-ras-)(P＞0.05). K-ras (12-Val) mutant peptide with the surface of cationic nanoparticle at lower concentrations can be effectively presenting on the surface of DCs than only K-ras (12-Val) mutant peptide.Conclusion:K-ras (12-Val) mutant peptide with cationic carrier can be effectively presenting and expression of DCs and induce CTL specific killing activity aganist pancreatic cancer cell lines with K-ras (12-Val) mutant peptide.

To explore the effect of coriaria lactone (CL)-activated astrocyte-conditioned medium on the cerebral TNF-alpha of normal rats, the CL-activated astrocyte-conditioned medium (ACM) was injected into the lateral ventricle of SD rats. The rats were observed for behavioral changes, and the changes of the expression of TNF-alpha in the cerebral cortex and hippocampus were immunohistochemically examined by employing SP method. TNF-alpha level was assessed by means of radioimmunoassay in homogenate of cerebral cortex and hippocampus as well as cerebrospinal fluid. Seizure episodes were observed in ACM group 30 min after the ACM injection, but they were not observed in the control group. Immunohistochemical detection showed that the immunoreaction of TNF-alpha in hippocampus and cerebral cortex of rats were stronger than that of the control group 4 h after the ACM injection (P<0. 05). In this group, the concentrations of TNF-alpha in homogenate of cerebral cortex and hippocampus and cerebrospinal fluid were higher than those of the control group (P<0.05). It is suggested that the ACM activated by CL can enhance the expression of TNF-alpha in normal rats, and is related to 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.

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.