1.Comparison of pathological features and prognosis between orbital and systemic diffuse large B cell lymphoma
Chinese Journal of Experimental Ophthalmology 2014;32(10):916-920
Background Orbital diffuse large B cell lymphoma(DLBCL) is a kind of malignant lymphoma with higher morbidity,but the systematic study is difficult for the fewer cases and the lack of orbital DLBCL cell line.Objective This study was to compare the differences or similarities of histopathological characteristics between orbital DLBCL and systemic DLBCL,and to discuss whether systemic DLBCL cell line is available to in vitro research of orbit DLBCL.Methods The histopathological specimens were collected from 3 orbital DLBCL patients and 15 systemic DLBCL patients in Tianjin Eye Hospital and Tianjin Tumor Hospital from 2004 to 2009.The prognosis of the patients was followed-up,and the survival curve was drew.The histopathological examination of the specimens was performed using hematoxylin and eosin staining,and the expressions of CD20,CD79α,CD45RO,CD10,BCL-6,mum-1,Ki-67 and surviving in the specimens were detected by immunochemistry.The results mentioned above were analyzed and compared between orbital DLBCL and systemic DLBCL.Results The histopathological examination showed that the centroblastic type was the primary form in both orbital DLBCL and systemic DLBCL,according with the diagnosis of DLBCL.The differences in the positive expression rate of CD20,CD79α,CD45RO,Ki-67,survivin were not statistically significant between the two types of DLBCL (P =0.167,0.442,1.000,1.000,0.442).Immunochemistry revealed that 3 orbital DLBCL patients were germinal center B-cell-like (GCB),and 2 of the patients showed the positive expression for BCL-6 and mum-1 as well as absent expression for CD10,but the other patient presented the absent expression for BCL-6,mum-1 and CD10.In 15 systemic DLBCL patients,7 were GCB type,with the positive expression for CD10 in all the 7 patients and absent expression for BCL-6 in 6 patients.In addition,in 8 of non-GCB type,3 appeared to be absent expressed for CD10 and BCL-6,and 5 were positive expressed for BCL-6 and mum-1 and absent expressed for CD10.No significant difference was found in the survival duration between the orbital DLBCL and systemic DLBCL (P =0.067).Conclusions There exists no clinically significant difference in the pathological features,the expression of tumor cell markers and prognosis in both orbital and systemic DLBCL.These two DLBCLs appear to be similar in epidemiology and clinical staging,inferring that systemic DLBCL cell line is available in the in vitro study of orbital DLBCL.
2.Establishment of mouse model of orbital diffuse large B cell lymphoma
Chinese Journal of Experimental Ophthalmology 2015;33(11):981-984
Background Recently, the morbidity of orbital lymphoma increased gradually, and we have made deeper research in pathology,therapy and pathogenesis of the disease.There were few reports of mice model of orbital lymphoma up to now for its lower morbidity and culture difficulty.Objective This study was to establish a mouse model of orbital diffuse large B cell lymphoma (DLBCL) by injection of systemic DLBCL cell line pfeiffer.Methods Ten SPF BALB/c mice and 5 nod-SCID mice were radiated firstly using 137Cs,with the absorption dose 3.5 Gy in the BALB/c mice and 2.6 Gy in the nod-SCID mice,and then pfeiffer cells were intraobitally injected in 4eyes of BALB/c mice (orbital injection group) and suncutaneously injectied in 4 eyes of BALB/c mice and 4 eyes of nod-SCID mice (subcutaneous injection group) at the concentration of 1.5×l08/ml.The developing status of tumors were examined once per day and the growth curve was drawn.The tumors and nearby lymph nodes were obtained 54 days after injection for the preparation of 4 μm thickness of serial sections.Hemotoxylin-eosin staining was used to examine the histopathology of the tumors, and immunochemistry was employed to detect the expressions of CD20,CD79α and CD45RO proteins.The tumors were typed based on the expressions of CD10, BCL-6 and mum-1 in the specimens,and the expressions of Ki-67 and survivin were assyed to assess the prognosis of the tumor.All the results were compared with 3 diagnosed human orbital DLBCL sections.The use and care of the mice complied with Chinese Administration Rule of Laboratory Animal.Results The tumor formation rates were 100% in both the orbital injection group and subcutaneous injection group, and the tumors grew much faster in nod-SCID mice than BALB/c mice.Infiltration of tumor cells in lymph nodes were found in the subcutaneous injection group rather than the orbital injection group.The pathological features were accordant among the orbital injection group, subcutaneous injection group and human orbital DLBCL sections.The number of <50% and ≥50% CD20+ specimens was 3 and 5,CD79αwas 2 and 6,CD45RO was 8 and 0 in the BALB/c mice;while that in the nod-SCID mice was 1 and 3 in CD20,0 and 4 in CD79α,4 and 0 in CD45RO;the number of human orbital DLBCL specimens was 1 and 2 in CD20,1 and 2 in CD79α,2 and 1 in CD45RO,without significant differences among them (all at P=1.00).No significant differences were seen in Ki-67+ number and survivin+ number among the BALB/c mice, nod-SCID mice and human orbital DLBCL specimens (all at P=1.00).The detection of CD10,BCL-6 and mum-1 expressions indicated that the tumors of BALB/c mice,nod-SCID mice and human orbital DLBCL specimens all were the non-germinal center B cell-like types.Conclusions The orbital and subcutaneous DLBCL mouse models are successfully established by injection of pfeiffer cell line.There are the same findings and features in biological behavior, pathology and immunohistochemistry in orbital,subcutaneous models with human orbitl DLBCL.Nod-SCID mice appear to be more suitable for the growth of lymphoma cells.
3.Impact of X-ray radiation on cell cycle and cell apoptosis of diffuse large B-cell lymphoma
Hong, JI ; Zhenfeng, GUO ; Fengyuan, SUN
Chinese Journal of Experimental Ophthalmology 2014;32(10):902-905
Background The malignant degree of the diffuse large B-cell lymphoma (DLBCL) is high.Radiation therapy is sensitive,but the therapeutic schedule can not be unified,reasonable and effective therapeutic schedule is important in clinic treatment of the DLBCL.Objective This study was to access the effect of X-ray on the cell cycle and cell apoptosis of the DLBCL.Methods DLBCL cell line was cultured and the cell suspension was inoculated to 12-well cultured plate.The cells exposed to X-ray for 6,12,24,48 hours with the irradiation doses of 2,4,6,8 Gy respectively.The cell proportions of different cycles were assayed by flow cytometer,and cell apoptosis index was evaluated and calculated.Non-irradiation cells served as controls.Results The cells grew well with a good cell vitality and round-like shape,and the growth was stable in generation 2 or 3.With the increase of X-ray doses and the lapse of irradiation time,the cell proportions in G0-G1 phase gradually reduced (Fdose =45.58,P=0.00; Ftime =43.11,P=0.00).However,cell proportions in G2-M phase were gradually increased as the increase of X-ray doses and lapse of irradiation time (Fdose =40.77,P =0.00 ; Ftime =91.91,P =0.00).In addition,the apoptotic proportion of the cells was significantly elevated with the increase of X-ray dose and irradiation time (Fdose =87.36,P =0.00; Ftime =37.31,P =0.00).Conclusions X-ray irradiation retards DLBCL in the G2-M phase and causes cell apoptosis in dose-and time-dependent manner.
4.Value of ultrasound and color Doppler imaging in children orbital tumors
Tong, WU ; Fengyuan SUN ; Dongrun, TANG
Chinese Ophthalmic Research 2009;27(11):1001-1004
Objective Imageology diagnosis of orbital tumor is very critical for the determination of nature of tumor in childhood patients. It is very necessary to comprehensively assess the results of clinical manifestations and imageology for the identification of orbital tumor. This study was to evaluate the clinical value of ultrasonography and color Doppler imaging (CDI) in the diagnosis of orbital tumors in children. Methods The clinical data, B-type echograms and CDI of 39 children with orbital tumors including capillary hemangioma, lymphangioma, venous hemangioma, rhabdomyosarcoma, dermoid cyst, varicosity etc. Were analyzed retrospectively. Results Among 39 cases of orbital tumors in children, 8 cases of capillary hemangioma and 2 cases of rhabdomyosarcoma showed the signals of artery blood flow from CDI ultrasound examination. No blood flow signal was obtained in 12 cases with dermoid and 8 cases with lymphangioma. The venous blood flow signals were examined from CDI examination in 3 out of 6 cases of venous hemangioma. Dynamic blood flow changes were seen from CDI in 3 cases with varicosis. Conclusion Combination of B-type ultrasonography and CDI is useful for the diagnosis of orbital tumors in childhood patients.
5.The expression of GST-π protein in peripheral blood and brain of patients with drug-resistant epilepsy and refractory epilepsy rats
Zhiqing SUN ; Qingxi FU ; Lixiang TU ; Fengyuan CHE ; Quanping SU
Chinese Journal of Nervous and Mental Diseases 2016;42(2):80-84
Objective To investigate the expression of glutathione S-transferase π (Glutathione S-transferase π, GST-π) protein in peripheral blood and brain of patients with drug-resistant epilepsy and refractory epilepsy rats. Meth?ods From January 2010 to March 2014, the expression of GST-πin the blood and brain of 32 cases of drug-resistant epi?lepsy underwent neurosurgery and 10 cases of cerebral vascular malformation underwent surgery were studied and com?pared. The expression of GST-πin the blood and brain in refractory epilepsy rats and normal rats were studied and com?pared. Results The specimen from 20 temporal, 6 frontal and 6 occipital lobes were obtained from drug-resistant epilep?sy patients. The expression levels of GST-πin the blood and brain in refractory epilepsy rats and normal rats were higher than those of the control groups (P<0.05). Conclusion GST-πmay be involved in the process of drug-resistant epilepsy. The GST-πexpression in blood may be used as a marker for resistance to anti-epileptic agents.
6.Progress in pathogenesis and treatment of the elevated intraocular pressure resulted from thyroid associated ophthalmopathy
Zhenfeng GUO ; Liang ZHAO ; Fengyuan SUN
Chinese Journal of Experimental Ophthalmology 2021;39(7):661-664
In adult orbital diseases, thyroid associated ophthalmopathy (TAO) is of very high incidence rate, and it can seriously affect patients' appearance, eyesight and binocular visual function and so on, and significantly reduce the patients' quality of life.In addition to the common manifestations such as eyelid retraction, exophthalmos and strabismus, some TAO patients may suffer from obviously increased ocular pressure and even visual field damage, which are often ignored or missed in diagnosis and should be paid more attention to.The pathogenesis of elevated intraocular pressure in TAO is mainly related to the elevated episcleral venous pressure and the extraocular muscles.Because the elevated intraocular pressure resulted from TAO is secondary and its pathogenesis is complex, personalized treatment different from primary glaucoma therapy is needed.In this article, the epidemiology, pathogenesis and therapy of elevated intraocular pressure in TAO including medication, surgery, and radiotherapy were reviewed to provide reference for the clinical diagnosis and treatment for TAO.
7.Protective effects of human induced pluripotent stem cell-derived exosomes on high glucose-induced injury in human endothelial cells
Qianqian DING ; Ruiting SUN ; Pingping WANG ; Heng ZHANG ; Meng XIANG ; Dan MENG ; Ning SUN ; Fengyuan CHEN ; Sifeng CHEN
Chinese Journal of Pathophysiology 2016;32(8):1530-1531,1532
AIM:To investigate the effects of human induced pluripotent stem cell-derived exosomes (hiPSC-exo) on cell viability, capillary-like structure formation , and senescence in endothelial cells exposed to high glucose .METHODS: Exosomes were isolated from the conditional medium of hiPSCs and confirmed by transmission electron microscopy , nanoparticle tracking analysis , and Western blot analysis using Alix and CD63 as markers.hiPSC-exo were labeled with PKH26 for tracking.Cultured HUVECs were treated with high glucose (33 mmol/L) with or without hiPSC-exo (20 mg/L) for 48 h, and cell viability, capillary tube formation, and senescence were assessed .RESULTS:hiPSC-exo showed a typical cup shape and could be taken up by human umbilical vascular endothelial cells (HUVECs) in a concentration-dependent manner.When exposed to high glucose, viability and tube formation in HUVECs was signifi-cantly reduced, whereas the proportion of senescent cells was higher compared to that in control HUVECs (P<0.01).Furthermore, hiPSC-exo restored cell viability and capillary-like structure formation , and reduced senescence in HUVECs exposed to high glucose (P<0.01).However, hiPSC-exo had minimal effects on normal HUVECs.Therefore, stem cell-derived exosomes can promote cell proliferation, enhance capillary-like structure formation , and reduce senescence in endothelial cells exposed to high glucose . CONCLUSION:Our study highlights the role of exosomes derived from hiPSC and may provide a new strategy for maintaining vascular health, preventing vascular aging , and avoiding pathological vascular remodeling that occurs in many diseases .
8.Advances in the research of cell electrofusion under microgravity.
Yan SUN ; Yanhong YUAN ; Zongchun YI ; Fengyuan ZHUANG ; Yubo FAN
Journal of Biomedical Engineering 2008;25(3):720-723
Bioseperation, cell cultivation and cell electrofusion are three main biological processes in space laboratories. Microgravity is free from the influences of convection and sedimentation. Therefore, it is an ideal realm for cell electrofusion and hence it can be used in the research of monoclonal antibody, cross breeding and microgravity biology. This paper reviews the research of cell electrofusion under microgravity, including the changes of cytoskeleton and the mechanism of cell electrofusion.
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Electric Stimulation
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Weightlessness Simulation
9.Changes of Muscle-related Genes and Proteins After Spaceflight in Caenorhabditis elegans
Chi WANG ; Chen SANG ; Higashibata AKIRA ; Ishioka NORIAKI ; Long RONG ; Chun YANG ; Yan SUN ; Zongchun YI ; Fengyuan ZHUANG
Progress in Biochemistry and Biophysics 2008;35(10):1195-1201
The molecular mechanism underlying muscular atrophy and gravisensing during spaceflight is still unknown. The major effects of spaceflight on body-wall muscles of Caenorhabditis elegans (C. elegans) in the structures and functions wore examined, and five important muscle-related genes and three proteins were studied after nearly 15-day spaceflight. The changes for the wall-muscles were observed in situ. Decreased muscle fiber size was observed with myosin immunofluorescence and duller dense-body staining in flight samples, which suggested that muscular atrophy had happened during spaceflight. However, F-actin staining showed no differences between the spaceflight group and ground control group. Otherwise, after returning to the earth the C eleganu displayed reduced rate of movement with a lower ratio (height/width) in crawl trace wave, which indicated a functional defect. These results demonstrated that C. elegans muscular development was changed in response to microgravity, and changes also occurred at the level of gene transcription and protein translation. Expression of dys-I increased significantly in body-wall muscles, while hlh-1, myo-3, uric-54 and eg1-19 RNA levels decreased after spaceflight. Dystrophin (encoded by dys-1) is one of important components in dystrophin-glycoprotein complex (DGC). Increased dys-I expression after flight implied that the muscular cell would accept more gravity signals by DGC in mierogravity in order to keep mechanical balance within the cells. It is concluded that DGC was involved into the mechanical transduction in body-wall muscles of C. elegans when gravity varied, which potentially played a vital role in gravisensing. The changes ofhlh-l, myo-3, tmc-54 and egl-19 suggested that they had the effects of promoting microgravity-induced muscular atrophy in strcture and function aspects. Result of Western blotting showed that the level of myosin A in spaceflight group decreased, further confirmed that atrophy happened during flight.
10.Advances in the study of exosomes in ophthalmic applications
International Eye Science 2024;24(4):596-600
Exosomes are commonly found in blood, urine, saliva, ascites, amniotic fluid and other body fluids, and are involved in intercellular communication, signal transduction, transport of genetic material, maintenance of internal environmental homeostasis and immune regulation, with a wide range of important biological functions. Exosomes transport proteins, lipids, and nucleic acids to target cells and facilitate intercellular communication.As research continues, they have been found to play important roles in physiological and pathological processes, and are important biomarkers for the diagnosis and treatment of diseases. It plays an important role in immunomodulation, inflammatory response, and angiogenesis in many diseases such as cancer, cardiovascular diseases, and brain diseases. More researches suggest that exosomes also play an important role in the development and progression of ophthalmic diseases. In this review, the research history and biological functions of exosomes, as well as their pathogenesis and prospects for the application in ophthalmic diseases, including corneal diseases, glaucoma, ocular trauma, age-related macular degeneration, uveitis and intraocular tumors, were discussed