3.The killing effect of NY-ESO-1-sensitzed dendritic cells-induced cellular toxic lymphocyte on human retinoblastoma cell
Chinese Journal of Experimental Ophthalmology 2012;30(7):586-591
Background Cell immunologic therapy for retinoblastoma(RB)is becoming a hot research topic.Cancer-testis antigen is a human immunogenic protein and is used to treat some tumors.However,its effect on RB has not been investigated.Objective The present study was to discuss the antigen specific anti-tumor effect of cytotoxic T lymphocytes(CTL)induced by the cancer-testis antigen,NY-ESO-1-sensitized dendritic cells(DCs),on human RB.Methods PCR was performed to amplify target gene fragments from the NY-ESO-1 plasmids,and then the target gene fragments were digested with the restriction enzymes SalI and EcoRI.Harvested fragments were inserted into the pDC316 plasmid to construct the recombinant plasmid pDC316/NY-ESO-1.The expression of NY-ESO-1 protein in human RB cells strain,HXO-RB44,was detected by immunofluorescence and Western blot.Monocytes were isolated from 60 ml of peripheral blood from a healthy donor using Ficoll density-gradient centrifugation with a cell density of 1 × 107/ml.DCs isolated from blood were stimulated with recombinant human granulocyte-macrophage colony stimulating factor(rhGM-CSF)and recombinant human interleukin-4(rhIL-4).The recombinant plasmid pDC316/NY-ESO-1 was transfected into DCs and the DCs were co-cultured with T lymphocytes.The resultant CTL were used as effector cells.The growth of the CTL was detected by MTT assay.The CTL were then added into the growth medium used for culturing HXO-RB44 cells and the vitality of the HXO-RB44 cells was assayed by MTT assay.Results The sequence of the cloned DNA fragment of the recombinant plasmid pDC316/NY-ESO-1 was conforms with the sequence of the NY-ESO-1 gene.The expression of the NY-ESO-1 protein in HXO-RB44 cells was tested by immunofluorescence and Western blot.DCs were successfully induced with rhIL-4 and rhGM-CSF from PBMC.The recombinant expression plasmid pDC316/NY-ESO-1 was successfully transferred into DCs.These DCs had high expression of surface molecules such as HLA-DR(42.1%),CD80(54.2%),CD83(39.7%)and CD86 (94.8%).The CTL that was induced by DCs-sensitized with NY-ESO-1 specifically killed HXO-RB44 cells.CTL induced by the sensitized DCs had a stronger cytotoxic effect against HXO-RB44 cells compared with un-sensitized DCs and CTL un-induced with DCs,as shown by MTT asssay(P<0.05).The anti-tumor activity was highest when the ratio of effector to target was 75∶1(P<0.05).Conclusions DCs transfected by the recombinant plasmid pDC316/NY-ESO-1 can induce the proliferation of allogenic CTLs,which showed a specific anti-tumor effect against HXO-RB44 cells.These results present a new type of immunotherapy for the treatment of RB.
4.Primary mucosal melanoma of the nasal cavity: a linicopathological analysis of 17 cases
Wen LIU ; Zhi ZENG ; Heng ZHOU ; Hao WU ; Jingping YUAN
Chinese Journal of Clinical and Experimental Pathology 2017;33(2):169-173
Purpose To study the clinicopathologic features and differential diagnosis of primary mucosal melanoma of the nasal cavity (PMMNC).Methods 17 cases of PMMNC diagnosed from January 2003 to September 2016 were studied by clinical pathological analysis and immunohistochemical staining,and relevant literatures were reviewed.Results 73% of the PMMNC was characterized by unilateral nasal congestion and intermittent epistaxis and 61% of the PMMNC occurred in the nasal septum and nasal side wall.Microscopically,the organizational structure and morphology were complex and diverse,which had several cell types including epithelioid cell type (6cases,35.3%),spindle cell type (3 cases,17.6%) and snall cell type (5 cases,29.4%),the other 3 cases (17.6%)were mixed cell type.Mitotic activity and tumor necrosis were more likely to be seen in PMMNC,among other clinicopathological features with a small amount of fibrous stroma and melanoma and rich blood vessels.The immunohistochemical study showed that the positive rate of S-100 and HMB-45 were both 93.8%(15 cases) and those of Melan-A and vimentin were both 87.5% (14 cases),while CK and EMA were both negative (16 cases).Conclusion PMMNC is a rare disease and the phenotype of S-100,HMB-45,Melan-A and vimentin are useful for diagnosis of PMMNC.
5.Study on interactions between targeted ultrasound contrast agents with anti-human interleukin-8 monoclone antibody and activated vascular endothelial cellls
Wei YANG ; Yongping LU ; Yuan YUAN ; Hong ZHOU ; Rui LIU ; Zhi JING ; Yueyue TANG
Chinese Journal of Ultrasonography 2012;21(8):715-718
Objective To observe the interactions between a anti-interleukin-8 monoclonal antibody (anti-IL-8 mAb) carried targeted ultrasound contrast agents and the injured vascular endothelial cells,as well as to explore the role of IL-8 in the formation of atherosclerotic plaques and a new assessing method of vascular endothelial functions.Methods The targeted ultrasound contrast agent was prepared by using a erosslinking agent to couple a anti-IL-8 mAb with SonoVue microbubbles.The interactions between SonoVue microbubbles/the targeted microbubbles and normal/injured endothelial cells were observed under an inverted microscope,respectively.The numbers of endothelial cells and adhered microbubbles were counted under high power magnification.The ratio of microbubbles to endothelial cells was calculated for the quantitative analysis of the interactions.Results In the control group,only a slight amount of original SonoVue microbubbles were bound to normal/injured endothelial cells.In contrast,it was visible under the microscope that the anti-IL-8 mAb carried SonoVue could be bound to endothelial cells,and the number of microbubbles bound to the surface of injured endothelial cells was significantly higher than that bound to the normal endothelial cells.Conclusions The anti-IL-8 mAb carried targeted ultrasound contrast agent could be readily bound to the surface of the injured cells specifically,and thus suggesting a new direction for ultrasonic detection of vascular endothelial injury and the ultrasonic assessment of vascular endothelial functions.
6.Pituitary carcinoma: report of a case.
Jing ZHOU ; Nan-yun LI ; Zhi-qiang ZHANG ; Chi-yuan MA ; Bo YU ; Hang-bo ZHOU
Chinese Journal of Pathology 2013;42(2):123-125
Adenoma
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pathology
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Brain Neoplasms
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secondary
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Chromogranin A
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metabolism
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Diagnosis, Differential
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Follow-Up Studies
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Humans
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Magnetic Resonance Imaging
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Male
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Middle Aged
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Neoplasm Recurrence, Local
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Pituitary Neoplasms
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diagnosis
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metabolism
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pathology
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surgery
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Reoperation
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Synaptophysin
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metabolism
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Temporal Lobe
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pathology
7.Antitussive constituents of Disporum cantoniense.
Xiu-Hai GAN ; Chao ZHAO ; Zhi-Yuan LIANG ; Xiao-Jian GONG ; Hua-Guo CHEN ; Xin ZHOU
China Journal of Chinese Materia Medica 2013;38(23):4099-4103
The antitussive activity assay for the root extraction of Disporum cantoniense was carried out with coughing mice induced by ammonia liquor. The results showed that the ethanol and water extractions of D. cantoniense possess strong antitussive activity, and the high dose of the former was better than positive control, and then the constituents of the ethanol extraction were separated and purified by various modern chromatographic techniques. Their structures were identified by physico-chemical properties and spectroscopic data. As a result, eight compounds were isolated and identified as stigmast-4-en-3-one(1), (22E, 24R)-ergosta-5, 7, 22-trien-3beta-ol(2), obtucarbamate A(3), obtucarbamate B(4), neotigogenin(5), azo-2, 2'-bis[Z-(2,3-dihydroxy-4-methyl-5-methoxy) phenyl ethylene] (6),dimethyl {[carbonylbis (azanediyl)] bis( 2-methyl-5, 1-phenylene) j dicarbamate (7) , and quercetin-3-O-pB-D-glucopyranoside(8). All compounds were isolated from this plant for the first time, and the result of bioactivity-directed isolation showed that compounds 3, 4, and 6 had obvious effect on antitussive activity, and compound 6 had the same level as positive control.
Animals
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Antitussive Agents
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chemistry
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isolation & purification
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pharmacology
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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pharmacology
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Ethanol
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chemistry
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Female
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Liliaceae
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chemistry
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Male
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Mice
10.Studies on neuronal tracing with pseudorabies virus.
Bi LI ; Ling ZHU ; Yuan-Cheng ZHOU ; Wan-Zhu GUO ; Zhi-Wen XU
Chinese Journal of Virology 2014;30(3):333-337
With its abilities of trans-synaptic tracing and self-replication and wide host range, pseudorabies virus (PRV) has been applied in the field of neuroanatomy since the 1970s. Four decades of PRV application have made many advances in researches on neuronal tracing with PRV. Mechanism studies focused on investigating infection of primary neurons and tracing direction in secondary neurons, while application studies focused on development of new pathological strains and innovation of tracing techniques. To date, the mechanism and application of viral tracing are not completely figured out yet. Integration of molecular biology technology will improve the efficiency in related researches.
Animals
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Cell Tracking
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Herpesvirus 1, Suid
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genetics
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physiology
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Humans
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Neurons
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virology
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Pseudorabies
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virology