ObjectiveTo establish and evaluate the methods which can quantify the ocular hemodynamics by combined scanning laser ophthalmoscopy video fluorescein angiography with computerized digital image processing system．MethodsAfter acquiring serial digital images by video capture from some angiography videotapes and measuring the gray values of some fixed areas over time in these serial images，the gray value curves and some parameters were obtained．The inter- and intra-observer reproducibility surveys were carried out．ResultsThe hemodynamic parameters were obtained，which included sectional choroidal filling time and rate，sectional papillary filling time and rate，retinal arterial and venous filling time and rate，arterovenous passing time and diameter of artery and vein．The intra- and inter-observer reproducibility were fairly good．Conclusioncomputerized digital image processing system is useful for understanding retinal and choroidal circulation．

Adult ; Chemical and Drug Induced Liver Injury ; Dimethylformamide ; poisoning ; Humans ; Liver ; pathology ; Liver Diseases ; blood ; pathology ; Male ; Necrosis ; Occupational Diseases ; therapy ; Poisoning ; pathology ; therapy

Age Factors ; Cardiopulmonary Bypass ; Child ; Child, Preschool ; Humans ; Infant ; Infant, Newborn ; P-Selectin ; blood ; Platelet Glycoprotein GPIb-IX Complex ; analysis

Many gene therapy protocols can induce antitumor immunity, however, the ex vivo approach restricts their uses. This sutydy intended to induce antitumor immunity by direct transfer of MHC class Ⅱ gene in vivo. Methods: MHC class Ⅱ gene cDNA was introduced directly into two tumors: P815, (a murine weak immunogenic mas-tocytoma) and B16 (a murine nonirnmunogenic melanoma) to observe the survival rate of the mice. Results: Tumori-genicity of P815 was reduced when MHC class Ⅱ gene was introduced directly into tumors in vivo. Further more, most vaccinated mice could survive after second challenge of P815. Co-injection of MHC class Ⅱ and B7 genes in the B16 also resulted in the tumor grow slowly, while the injection of MHC class Ⅱ gene was not enough to induce effective antitumor responses. Conclusion: The results showed the potential applications of direct transfer of MHC class Ⅱ gene in the treatment of tumor.

Objective To construct the multiple antigenic peptide (MAP) gene and E. coli ex-pression system, based on the out membrane protein OmpL1, LipL21 and LipL32 from Leptospira interro-gans, and better understanding of the immunological activity of the recombinant protein. Methods Using MI3KE display and Western blot, the advantage epitopes of OmpL1, LipL21 and LipL32 were identified and used to synthesize a new gene, then its prokaryotic expression system was constructed. The expression of re-combinant protein was determined by SDS-PAGE. The immunity activity of the recombinant protein was iden-tified by Western blot and ELISA. Results The synthetic gene was effectively expressed in E. coli and mainly presented in soluble form. The expression protein could react with the antileptospirosis antibodies in rabbit and human sera, which contained different serogroups. Conclusion The recombinant MAP gene of leptospires was successfully constructedand and expressed in E. coli. The recombinant protein had a good immune activity, and could cross-reacted with antileptospirosis antibodies from different serogroups.

The particularities of oil suspension formulation can raise the invasive rate of Hirsutella sinensis to host of Hepialidae for the commercialization of artificial cultivation of Cordyceps sinensis. So it is important to develop a high cell viability oil suspension formulation of C. sinensis spawn. According to the characteristics of the oil suspension formulation, MTT assay is adapted and optimized. The result is as follows: reaction time 120 min, reaction temperature 37?C, methylbenzene as extracting agent, and a positive linear correlation established between active cell weights and cell viability. Varieties and concentrations of assistance agents in oil suspension formulation have been selected with the refined MTT assay, and further optimized together with cell concentrations through orthogonal experiment. The optimal combination project was obtained, namely, cell concentration 0.15 g/mL, aluminium stearate 60 mg/mL, and SPAN-80 50 ?L/mL. Results of stability test on the oil suspension formulation indicate that cell viability can maintain above 90% at 4?C after one month.

The generation of large quantities of novel human T cell clones ex vivo would make a wide range of gene-and immuno-therapies for tumor and AIDS possibly. Although it is well established that T cells are derived from CD34(+) cells, the involvement of thymic fragments from either human or murine fetus makes the in vitro T cell perliferation very cumbersome. In this report, cord blood mononuclear cells were used as accessory cells to support the differentiation of CD34(+) cells into naive T cells stimulated with SCF and IL-2. CD4(+) and CD8(+) T cells, under the cultural conditions, were continuously produced in vitro at least over a period of 3 weeks and their ratios changed gradually. CD4/CD8 double positive T cells and RAG-2 gene were existed, and RAG-2 gene, reponsible for TCR rearrangement, was expressed during the cell proliferation. Our study presents a simple culture system in vitro to acquire large quantities of naive T cell clones.

Objective To investigate the inhibitory effects ofPolyphyllin D (Paris saponin [PS]Ⅰ) and formosanin C (PS Ⅱ) on human glioblastoma U251 cells and its mechanism.Methods Glioma cell line U251 was cultured in vitro.Different treatments (PSⅠ and PS Ⅱ) were added into the medium cultured human malignant glioma U251 cells; and blank control group was also established.The effects of PSⅠ and PS Ⅱ on proliferation of glioma cells U251 in vitro was examined using methyl thiazolyl tetrazolium (MTT) assay.The nuclear morphology changes of apoptotic cells were detected by Hoechst33258 fluorescent staining.Apoptotic rate was quantified by flow cytometry (FCM) using Annexin-V/PⅠ dual staining.Western blotting was used to evaluate the changes of Fas,Caspase-8 and Caspase-3 proteins in U251 cells.Results PS Ⅰ and PS Ⅱ inhibted U251 cell proliferation with a timeand dose-dependent manner; as compared with that in the blank control group,the proliferation rate in the PSⅠ and PS Ⅱ treatment groups was significantly increased (P＜0.05); PS Ⅰ had stronger inhibited effect than PS Ⅱ.Typical morphological changes of apoptosis were observed in U251 cells of the PSⅠ and PS Ⅱ treatment groups with Hoechst 33258 staining.The early apoptotic rates of the PSⅠ and PS Ⅱ treatment groups were all significantly higher than that of the blank control groups (F=81.434,P=0.000).As compared with those in the blank control group,the expressions ofFas,Caspase-8 and Caspase-3 were significantly increased (P＜0.05).Conclusion PSⅠ and PS Ⅱ inhibite the proliferation ofglioma cells in vitro; they increase the expressions ofFas,Caspase-8 and Caspase-3,and accelerate the cell apoptosis,which might be the mechanism of inhibition.

Adenocarcinoma ; pathology ; Ciliary Body ; Humans ; Iris Neoplasms ; pathology ; Lung Neoplasms ; pathology ; Male ; Middle Aged ; Uveal Neoplasms ; pathology

OBJECTIVETo study the mechanism underlying the effect of combined use of cyclonpamine and hydroxycamptothecin in inducing the apoptosis of human oral squamous cell carcinoma cell line (OSCC) HSQ-89.

METHODSCCK8 assay was used to investigate the inhibitory effect of cyclopamine on HSQ-89 cells. Flow cytometry (FCM) was employed to examine the cell apoptosis following combined treatment with cyclonpamine and hydroxycamptothecin. Reverse transcription polymerase chain reaction (RT-PCR) was applied to detect the mRNA expressions of Bcl-2, Bcl-xl, and Bid in HSQ-89 cells after the treatments.

RESULTSCombined treatment with cyclonpamine and hydroxycamptothecin significantly inhibited the cell proliferation compared with hydroxycamptothecin treatment alone, also resulting in a significantly higher apoptosis rate of the cells (P<0.05). The mRNA level of Bcl-2 was significantly decreased after the treatments, especially after the combined treatment. Cyclopamine produced no significant effect on the mRNA levels of Bcl-xl and Bid in the cells.

CONCLUSIONThe combined use of cyclopamine and hydroxycamptothecin significantly down-regulates the expression on of bcl-2 to induce the apoptosis of human OSCC cell line HSQ-89.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Camptothecin ; analogs & derivatives ; pharmacology ; Carcinoma, Squamous Cell ; pathology ; Cell Line, Tumor ; Drug Synergism ; Humans ; Mouth Neoplasms ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Veratrum Alkaloids ; pharmacology