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.

Whether transforming growth factor-beta2 (TGF-beta2) induces apoptosis of human trabecular meshwork cells was investigated in vitro. Cultured 3-5 passage human trabecular meshwork cells were treated with 0 (control), 0.32, 1, 3.2 ng/ml TGF-beta2 for 48 h and divided into control group and experimental group. The apoptosis of human trabecular meshwork cells was examined by transmission electron microscopy, TUNEL technique and flow cytometry. The results showed characteristic morphologic changes of apoptotic cells were observed under transmission electron microscopy. DNA fragmentation of human trabecular meshwork cells was found by TUNEL technique. Quantitative analysis of flow cytometry showed that percentages of apoptotic human trabecular meshwork cells were (2.79 +/- 0.44)%, (4.43 +/- 1.17)% and (9.60 +/- 2.05)% respectively with different concentrations [1 ng/ml (P<0.05), 3.2 ng/ml (P<0.01)] of TGF-beta2 with the difference being significant between experimental group and control group [(1.41 +/- 0.34)%]. It was concluded that TGF-beta2 can induce apoptosis of human trabecular meshwork cells in vitro and may be involved in the decrease of trabecular meshwork cells in the patients with primary open angle glaucoma and aging of normal people.
Apoptosis/*drug effects ; Cells, Cultured ; Trabecular Meshwork/*cytology ; Transforming Growth Factor beta/*pharmacology ; Transforming Growth Factor beta2

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.

Whether cultured bovine trabecular meshwork cells and trabecular tissue ex vivo express insulin-like growth factor-I (IGF-I) messenger RNA (mRNA) and protein was investigated. Total RNA of cultured bovine trabecular meshwork cells as well as trabecular meshwork tissue freshly excised from bovine eyes was extracted, and reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect IGF-I mRNA. RT-PCR product was verified by sequencing. Immunohistochemical stain was used to detect IGF-I protein. The results showed that a single PCR amplified product was obtained, and the sequence was homologous to the known sequence.. IGF-I immunostain was positive in the cytoplasm of trabecular meshwork cells. It was concluded that trabecular meshwork cells produce IGF-I and contribute to the presence of IGF-I in trabecular meshwork microenvironment as well as aqueous humor. Trabecular meshwork cells were affected by IGF-I not only through paracrine, but also autocrine action. Whether abnormal down-regulations in IGF-I production may contribute to the pathogenesis of primary open-angle glaucoma and the possibility of promoting the autocrine action of IGF-I by trabecular meshwork cells to treat the disease is worth further investigation.
Cells, Cultured ; Cloning, Molecular ; Glaucoma, Open-Angle/etiology ; Glaucoma, Open-Angle/metabolism ; Insulin-Like Growth Factor I/biosynthesis ; Insulin-Like Growth Factor I/*genetics ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Trabecular Meshwork/cytology ; Trabecular Meshwork/*metabolism

Objective:To evaluate the role of autophagy in reduction of high glucose and hypoxia-reoxygenation (HG+ H/R) injury to isolated cardiomyocytes by dexmedetomidine in rats.Methods:The normally cultured rat H9c2 cardiomyocytes at the logarithmic growth phase were seeded in 6-well plates at a density of 1×10 6 cells/ml and divided into 4 groups ( n=15 each) using a random number table method: control group (group C), group HG+ H/R, dexmedetomidine group (group DEX) and dexmedetomidine+ autophagy inhibitor 3-methylpurine group (group 3-MA). The cells were incubated in culture medium with 1% fetal bovine serum + 1% double antibody for 24 h when the cell density reached 50%.To establish HG+ H/R injury model, the cardiomyocytes were cultured in high-glucose culture medium (glucose concentration of 33 mmol/L) for 24 h, and then incubated in a 37 ℃ incubator (95% N 2+ 5%CO 2) for 4 h followed by reoxygenation (90%O 2+ 10%CO 2) for 2 h. Dexmedetomidine was added until the final concentration reached 5 μmol/L at 1 h before hypoxia in DEX and 3-MA groups, and 3-MA was added until the final concentration reached 5 μmol/L at 1 h of incubation with dexmedetomidine.At 2 h after reoxygenation, the cell viability was recorded by the cell counting kit-8 assay, lactate dehydrogenase (LDH) activity was detected by LDH kit, the expression of autophagy-related protein LC3, P62 and Beclin-1 was detected by Western Blot, the ratio of LC3Ⅱ/LC3Ⅰwas calculated, and the expression of P62 and Beclin-1 mRNA was detected by real-time polymerase chain reaction. Results:Compared with group C, the cell viability was significantly decreased in HG+ H/R, DEX and 3-MA groups, LDH activity in the supernatant was increased and expression of P62 was decreased in HG+ H/R and 3-MA groups, ratio of LC3Ⅱ/LC3Ⅰwas decreased in group 3-MA, and the expression of Beclin-1 was down-regulated in group HG+ H/R ( P<0.05). Compared with HG+ H/R group, LDH activity in the supernatant was significantly decreased, expression of Beclin-1, P62 and its mRNA was up-regulated, and ratio of LC3Ⅱ/LC3Ⅰwas increased in DEX group, and LDH activity in the supernatant was increased in group 3-MA ( P<0.05). Compared with DEX group, cell viability were decreased, LDH activity in the supernatant was increased, Beclin-1, P62 and its mRNA was down-regulated, and ratio of LC3Ⅱ/ LC3Ⅰwas decreased in group 3-MA ( P<0.05). Conclusion:Autophagy is involved in the reduction of HG+ H/R injury to isolated cardiomyocytes by dexmedetomidine in rats.

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 localize and characterize the plasmid protein pORF5 in the Chlamydia trachomatis(Ct) infected cells. Methods The open reading frame encoding for pORF5 protein from the Ct plasmid was amplified and cloned into the pGEX-6p vector. The recombinant plasmid pGEX-pORF5 was transformed into XL1-blue E. coli to express fusion protein with the glutathione-s-transferase (GST). After purified with Glutathione Sepharose 4B beads, the pORF5 fusion protein was used to immunize mice to make monoclonal and polyclonal antibody. The antibodies were used to localize the endogenous pORF5 protein and detect the expression pattern in Chlamydia-infected cells using an indirect immunofluorescence assay (IFA). At the same time, ELISA was used to determine whether pORF5 plasmid protein was expressed and immunogenic during Ct infection in humans. Results pORF5 was detected a dominant signal in the cytosol of the Chlamydia-infected cells with a pattern similar to that of anti-CPAF. pORF5 also appeared in the RBs and EBs in small quantity. Athough pattern was similarly, pORF5 did not overlap with CPAF. pORF5 protein was strongly recognized antiserum in an ELISA. Conclusion The pORF5 plasmid protein was identified as a secreted protein with good immunogenicity, pORF5 gene was to express the endogenous target protein during human infection.