1.The pathogical characteritics and mechanism of craniocerebral injuries concomitant with high-velocity missile wound of max-illofacial region
Zumeng YA ; Zhongyu LI ; Yinghui TAN
Journal of Third Military Medical University 1984;0(01):-
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.
2.Iris pigment epithelial cells of rabbits stained by carboxyfluorescein diacetate succinimidyl ester in vitro
Shaomin PENG ; Zhongyu ZHANG ; Xiaoxin LI
Chinese Journal of Ocular Fundus Diseases 2000;0(04):-
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.
3.Effects of activated ACM on expression of signal transducers in cerebral cortical neurons of rats.
Xiaojing, WANG ; Zhengli, LI ; Changgeng, ZHU ; Zhongyu, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(3):230-2
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.
4.Expression, purification and identification of recombinant Omp~2 of Chlamydia trachomatis
Chaoqun CHEN ; Yimou WU ; Zhongyu LI ; Weiguo YIN ; Lizhi TAN
Chinese Journal of Laboratory Medicine 2003;0(07):-
Objective To express outer membrane protein 2(Omp2) of Chlamydia trachomatis, purify expressed products and study its immunity.Methods The target gene encoding Omp2 167—434 amino acid residues was amplified by PCR from C. trachomatis template DNA. The targeted DNA fragment was cloned into expression vector pET28b(+) and introduced into competent E. coli BL21(DE3) cell. Recombinant Omp2aa_ 167 ~aa_ 434 was expressed after induction by IPTG and analyzed by SDS-PAGE and Western blot, purified with Ni-NTA-His affinity chromatography. The rOmp2aa_ 167 ~aa_ 434 was used to immune rabbits for immunogenicity assessment.Results Restriction enzymes cleavage analysis and DNA sequencing confirmed that the plasmid pET28b(+)/Omp2aa_ 167 ~aa_ 434 was correctly constructed. The 35.0?103 molecular weight pure protein, which specifically reacted with serum from C. trachomatis infected patient by Western blot, was obtained by optimizing the conditions for both expression and purification. The titer of serum antibodies was above 1∶1 280 as detected by ELISA.Conclusion The expressed product showed good immunity.
5.Antitumor efficacy of specific cytotoxic T lymphocytes activiated by dendritic cells pulsed with K-ras antigen to pancreatic cancer ex vivo
Li XU ; Yanqiu FANG ; Guang TAN ; Zhongyu WANG ; Yan TAN
Chinese Journal of Immunology 2009;25(12):1075-1079,1091
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.
6.Study on Concentrations of Formaldehyde in Newly Decorated Apartments
Yuan GAO ; Zhongyu XU ; Li ZHAI ; Al ET ;
Journal of Environment and Health 1992;0(05):-
Objective To investigate the concentrations of formaldehyde in newly decorated apartments. Methods In this study we monitored the concentrations of formaldehyde in indoor air of 28 newly decorated apartments and offices and furnishings. In order to analysis the correlation between formaldehyde concentration of indoor air and personal exposure level, 17 persons' exposure levels were measured by personal air monitoring in 2 offices. Results The medians of formaldehyde concentrations in indoor air and furnishings were 0.062 mg/m3 and 0.260 mg/m3 respectively. The concentration of formaldehyde in furniture was significantly higher than that in indoor air (Z=-5.098,P
7.Apoptosis of human trabecular meshwork cells induced by transforming growth factor-beta2 in vitro.
Yang, CAO ; Houren, WEI ; Michael PFAFFL ; Banghong, DA ; Zhongyu, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(1):87-9, 94
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
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Cells, Cultured
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Trabecular Meshwork/*cytology
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Transforming Growth Factor beta/*pharmacology
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Transforming Growth Factor beta2
8.Effect of coriaria lactone-activated astrocyte-conditioned medium on the cerebral TNF-alpha of normal rats.
Zhongyu, LI ; Qingying, LIU ; Changgeng, ZHU ; Wei, WANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(2):161-3, 184
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.
9.Analysis of construction of teaching staff and excellent college teaching team
Zhongyu LI ; Yimou WU ; Cuiming ZHU ; Yan ZHANG
Chinese Journal of Medical Education Research 2006;0(08):-
Colleges are responsibile for training high-quality professionals and top talents.College teachers are decisive for teaching quality.As a result,to strengthen the faculty construction and establish excellent college teaching team are important for the improvement of the quality of personnel training.
10.Insulin-like growth factor-I gene cloning and protein expression in bovine trabecular meshwork tissue and cells.
Yang, CAO ; Houren, WEI ; Banghong, DA ; Michael, PFAFFL ; Zhongyu, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2002;22(1):69-72
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
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Cloning, Molecular
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Glaucoma, Open-Angle/etiology
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Glaucoma, Open-Angle/metabolism
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Insulin-Like Growth Factor I/biosynthesis
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Insulin-Like Growth Factor I/*genetics
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RNA, Messenger/biosynthesis
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RNA, Messenger/genetics
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Reverse Transcriptase Polymerase Chain Reaction
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Sequence Analysis, DNA
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Trabecular Meshwork/cytology
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Trabecular Meshwork/*metabolism