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.Trichostatin A Inhibits Proliferation and Induces Expression of p21WAF and p27 in Human Brain Tumor Cell Lines
Zhi-Min WANG ; Jin HU ; Dai ZHOU ; Zhi-Yuan XU ; C.Panasci LAWRENCE ; Zhong-Ping CHEN
Chinese Journal of Cancer 2002;21(10):1100-1105
Background and Objective: The histone deacetylase inhibitor,trichostatin A(TSA),was shown to induce apoptosis in transformed cells at submicromolar concentrations. However, the effect of TSA on brain tumor cells is still unknown. This study was designed to investigate whether TSA posses antitumor activity and if any, its mechanism. Materials and Methods: A p53 mutant human glioma cell line T98G and a p53 wild type human neuroblastoma cell line SKNSH were exposed to TSA. Cell proliferation was assessed by sulforhodamine B (SRB) cytotoxicity assay. Apoptosis was quantified by flow cytometry and confirmed by apoptotic ladder formation. Expression patterns of accumulation of highly acetylated histone H3,H4; p53 and cell cycle-associated p21waf,p27 which were induced by TSA were determined by using Western blot analysis. Results: TSA inhibited the proliferation of brain tumor cell lines at nanomolar concentrations and induced accumulation of highly acetylased histone moleculars. Treatment with TSA at 0.33μ M for 24h significantly induced cell apoptosis.In addition to the suppression of cell growth, the up regulation of p21waf and p27 expression was observed within 48h after the treatment.p21 protein levels were increased at early time points and reached maximal levels at 8h, while p27 protein levels were increased after 8h. However, there was no significant changes of acetylased p53 and endogenous p53 protein levels were observed. Conclusion:TSA may inhibit brain tumor cell growth in vitro, which is otherwise particularly resistant to chemotherapy. TSA acts as an anti-tumor agent could be through co-operation between p21 and p27 in growth inhibition, irrespective of endogenous p53 status.
8.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
9.Efficacy and safety of different hemostatic drugs in transurethral resection of the prostate in elderly patients
Qingquan LIU ; Zhi CHEN ; Danqing ZHOU ; Yin ZHANG ; Yuan CHEN ; Qianyuan ZHUANG ; Cuntai ZHANG
Chinese Journal of Geriatrics 2016;35(8):850-853
Objective To investigate the efficacy and safety of perioperative use of different hemostatic drugs in elderly patients undergoing transurethral resection of the prostate (TURP).Methods A total of 480 elderly patients with benign prostatic hyperplasia (BPH) admitted to the Departments of Urology and Gerontology of our hospital from January 2010 to October 2014 were selected and randomly divided into 6 groups (n =80,each).Patients in all groups were given preoperative oral finasteride for 1 week,with each of the non-control groups receiving perioperative heamocoagulase agkistrodon,mangabeys hemocoagulase,reptilase,ethylenediamine diaceturate,or dicynone + paraaminomethyl benzoic acid (PAMBA).Intraoperative bleeding,perioperative hematocrit,hemoglobin change,continuous bladder irrigation time,adverse reactions and other clinical parameters were recorded.Results Operations went smoothly with all patients.Changes in perioperative blood routine indexes,coagulation function,intraoperative bleeding volume and continuous bladder irrigation time had significant differences between the control group and the groups treated with hemostatic drugs (all P<0.05).No significant differences in perioperative blood routine indexes,coagulation function,intraoperative bleeding volume and continuous bladder irrigation time were found between groups treated with heamocoagulase agkistrodon,mangabeys hemocoagulase and reptilase,or between groups treated with ethylenediamine diaceturate and dicynone+PAMBA (all P> 0.05).There were significant differences in the perioperative blood routine indexes,coagulation function,intraoperative bleeding volume and continuous bladder irrigation time between the groups with and without blood coagulation enzymes (all P <0.05).Fibrinolytic responses occurred in 2 patients in the heamocoagulase agkistrodon group and were mitigated after drug withdrawal and fibrinogen infusion.Conclusions A reasonable perioperative dose of a hemostatic drug has good clinical effects in elderly patients undergoing TURP.Hemostatic drugs such as blood coagulation enzymes have certain advantages including safety over other types of hemostatic drugs,but their doses should not be too large.
10.Enhanced Effects of BoNT/A DNA Vaccines by Electric Pulses and Bupivacaine
Yun-Zhou YU ; Na LI ; Shuang WANG ; Wei-Yuan YU ; Zhi-Wei SUN ;
China Biotechnology 2006;0(05):-
Objective:To determine if suitable electric pulses-mediated DNA and DNA and bupivacaine complexes delivery technologies could enhance effects of botulinum neurotoxin serotype A (BoNT/A) DNA vaccines in mouse model. Methods:Vaccination of mice i.m. with plasmid DNA replicon vaccine pSCARSHc and conventional plasmid DNA vaccine pcDNASHc following electric pulses and with DNA and bupivacaine complexes. AHc-specific group antibody ELISA titers and lymphocyte proliferative responses of mice were detected and IgG1 and IgG2a isotype profiles were assayed. Results:Immune effects of DNA vaccines were enhanced following electric pulses and bupivacaine delivery. Effects of DNA vaccines following electric pulses were better than that of DNA vaccines formulated with bupivacaine,and the combined delivery technology of electric pulses and bupivacaine induced the highest level of specific antibodies and lymphocyte proliferative responses. Plasmid DNA replicon vaccine pSCARSHc induced relatively higher AHc-specific antibodies and lymphocyte proliferative responses in immunized Balb/c mice than conventional plasmid DNA vaccine pcDNASHc in these DNA delivery technologies. And vaccine pSCARSHc induced Th2/Th1-type immune responses with a general bias to Th2-type,and vaccine pcDNASHc induced Th2-type immune responses. Conclusion:Suitable electric pulses-mediated DNA and DNA and bupivacaine complexes delivery technologies could enhance effects of BoNT/A DNA vaccines in mouse model. Therefore,the methods described here potentially provide suitable strategies in developing an efficacious vaccine against botulinum neurotoxin serotype A.