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Objective:To investigate the effects of sclerostin (SOST) and WNT/CTNNB1 signaling pathway on the cell cycle, migration and invasion of human uveal melanoma (UM) cells and its related mechanism.Methods:UM tissues from 20 cases of epithelioid UM and 16 cases of spindle cell type UM were collected.The contents of SOST, Wnt-1 and Catenin beta-1 proteins in the collected tissues were detected by immunohistochemical staining.Three human UM tissue derived cell lines OCM-1 (primary spindle cell type), Mum-2B (metastatic epithelioid) and Mum-2C (metastatic spindle cell type) were selected and divided into three groups, blank control group not transfected, empty vector group transfected with SOST negative control vector and SOST siRNA group transfected with SOST siRNA.After 24-hour transfection, the mRNA and protein expression levels of SOST, CTNNB1, WNT protein family 1 (WNT1), CCND1, matrix metalloproteinase (MMP)2 and MMP9 were detected by real-time fluorescence quantitative PCR and Western blot, respectively.The invasion and migration ability of the transfected cells were measured by transwell method, and the cell cycle distribution was detected by flow cytometry.Another 9 female BALB/c nude mice were selected and randomized into OCM-1 group, OCM-1 empty vector group and SOST shRNA group, inoculated with OCM-1 without lentivirus infection, OCM-1 with blank lentivirus infection and OCM-1 with SOST shRNA lentivirus infection, respectively.Six weeks after inoculation, the in situ formation of tumor was observed.The interaction between SOST and low density lipoprotein receptor related protein(LRP)-5/6 in OCM-1 cells was explored by co-immunoprecipitation assay.The study protocol was approved by the Ethics Committee of Tianjin Medical University Eye Hospital (2018KY[L]-20).Results:Immunohistochemical staining results showed that the SOST expression level was higher and the expression levels of Wnt-1 and Catenin beta-1 were lower in spindle cell type UM tissues than in epithelioid UM tissues, and the differences were all statistically significant (all at P<0.01). The real-time fluorescence quantitative PCR results showed that the relative expression of SOST mRNA was significantly lower and the relative expressions of CCND1, WNT1 and MMP9 mRNA were significantly higher in SOST siRNA groups than in corresponding empty vector groups in the three cell lines (all at P<0.05). In OCM-1 and Mum-2C cell lines, the relative expressions of CTNNB1 mRNA were significantly higher in SOST siRNA groups than in empty vector groups (all at P<0.01). Western blot results showed that the relative expression of SOST protein was significantly lower and the relative expressions of Wnt-1, Catenin beta-1, cyclin-D1, MMP2 and MMP9 proteins were significantly higher in SOST siRNA groups than in empty vector groups (all at P<0.01). Transwell assay showed that the cell invasion and migration ability of SOST siRNA group was significantly higher than that of blank control group and empty vector group in the three cell lines (all at P<0.01). Flow cytometry showed that the proportion of G1-phase cells and the G1/S-phase ratio were significantly lower in SOST siRNA group than in blank control groups and empty vector groups (all at P<0.01). The eyeball volume of OCM-1 group, OCM-1 empty vector group and SOST shRNA group was (42.7±4.6), (49.0±22.9) and (135.2±32.7)mm 3, respectively, showing a significant overall difference ( F=19.963, P<0.01). The eyeball volume of SOST shRNA group was larger than that of OCM-1 group and OCM-1 empty vector group, and the differences were statistically significant (both at P<0.05). Co-immunoprecipitation results showed that SOST could interact with LRP-5 and LRP-6 by binding to them, respectively. Conclusions:Silencing SOST can promote the invasion and migration of UM cells, and increase the proportion of UM cells in the division phase.Silencing SOST can promote tumor growth in eyes of nude mice.SOST may play this function by interacting with the membrane receptor LRP-5/LRP-6 and then regulating the WNT/CTNNB1 signal pathway.
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Objective:To evaluate the influence of the clinical staging and different risk factors for the prognosis of ocular adnexal lymphoma.Methods:An ambispective cohort study was conducted.Seventy-four patients diagnosed with primary ocular adnexal lymphoma by pathology at Tianjin Medical University Eye Hospital from November 2010 to December 2018 were enrolled.TNM staging was performed according to local tumor extent, lymph node or systemic involvement.Ann Arbor staging was carried out according to lymph node involvement and extranodal extension.The pathological subtype was classified according to World Health Organization classification of lymphoma.The outcome of disease progression or death was analyzed.Kaplan-Meier method was used for univariate survival analysis.Cox proportional hazard model was employed for multivariate survival analysis to predict the risk factors affecting prognosis, hazard ratio ( HR) and 95% confidence interval ( CI) were estimated.This study adhered to the Declaration of Helsinki.The study protocol was approved by an Ethics Committee of Tianjin Medical University Eye Hospital (No.2021KY[L]-32). Written informed consent was obtained from all patients before entering the cohort. Results:For TNM staging, there were 68 cases in stage <T4, accounting for 91.9%, 6 cases in T4, accounting for 8.1%, 71 cases in N0, accounting for 95.9%, 3 cases in ≥N1, accounting for 4.1%, and no case was in stage M. For Ann Arbor staging, there were 72 cases in stage ⅠE, accounting for 97.3%, and 2 cases in stage ⅡE, accounting for 2.7%.As for pathological classification, 64 cases had mucosa-associated lymphoid tissue (MALT) lymphoma, accounting for 86.5% and 10 cases had non-MALT lymphoma, accounting for 13.5%.The follow-up of the patients was 3 to 117 months, with a median follow-up of 53 months.There were 6 cases dying of disease and 19 cases progressed.The 3-year and 5-year overall survival rates were 96.6% and 86.6%, respectively.The 3-year and 5-year progression-free survival rates were 75.6% and 65.9%, respectively.According to single-factor analysis, T4 stage, non-MALT type and Ki67 positive rate ≥10% were related to declined overall survival rate ( P<0.05). T4 stage, ≥N1 stage, ≥Ann Arbor Ⅱ stage, non-MALT type and Ki67 positive rate ≥10% were related to declined progression-free survival rate ( P<0.05). According to multiple-factor analysis, pathological type ( HR=33.193, 95% CI: 3.388-325.156, P=0.003) was the independent risk factor for overall survival rate.N stage ( HR=11.683, 95% CI: 2.720-50.173, P=0.001) and pathological type ( HR=11.337, 95% CI: 3.841-33.464, P<0.001) were independent risk factors for progression-free survival rate. Conclusions:TNM staging and pathological type are important clinical prognostic indicators for ocular adnexal lymphoma.Patients with high TNM stage or non-MALT lymphoma should be monitored closely.
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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.
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Objective:To explore the surgical outcomes and surgery-related complications of balanced orbital decompression and endoscopic transnasal inferomedial wall decompression for Grave's ophthalmopathy (GO).Methods:A cohort study was performed.The 56 GO patients who underwent balanced orbital decompression or endoscopic transnasal inferomedial wall decompression in the Tianjin Medical University Eye Hospital from December 2016 to December 2019 were enrolled.The follow-up time was 6 months.Patients were divided into two groups according to the operation modes.Thirty-three eyes of 24 subjects were given deep lateral wall rim-sparing orbital decompression and transcaruncular medial wall decompression, and 51 eyes of 36 cases were given endoscopic transnasal inferomedial wall decompression.The demographics, surgical details, imaging data, postoperative changes of exophthalmos, best corrected visual acuity (BCVA), orbital pressure and diplopia, surgery-related complications and further treatment were analyzed and compared.This study followed the Declaration of Helsinki and was approved by the Ethics Committee of Tianjin Medical University Eye Hospital [No.2020KY(L)-39]. All subjects signed informed consent.Results:The exophthalmos was (21.03±3.11)mm before operation, which was significantly higher than (17.06±2.55)mm after operation in the balanced orbital decompression group ( P<0.05). The exophthalmos was (20.51±3.53)mm before operation, which was significantly higher than (16.28±2.96)mm after operation in the endoscopic transnasal inferomedial wall decompression group ( P<0.05). No significant difference in the mean reduction of proptosis was found between the two groups ( P>0.05). All the subjects were accompanied with increase of intraorbital pressure before operation.The intraorbital pressure returned to normal at 6 months after operation.The postoperative BCVA of subjects with dysthyroid optic neuropathy (DON) were significantly higher than preoperative values ( Z=-3.524, -4.376; both at P<0.01). The postoperative improvement values of BCVA were 0.48 (0.25, 0.67) and 0.72 (0.40, 0.80) in the balanced orbital decompression group and the endoscopic transnasal inferomedial wall decompression group, respectively, with a significant difference between the two groups ( Z=-2.481, P=0.016). The incidence of complications in the balanced orbital decompression group was 21.2% (7/33), which was significantly lower than 47.0% (24/51) in the endoscopic transnasal inferomedial wall decompression group ( χ2=5.748, P=0.017). Conclusion:The two kinds surgical methods can effectively reduce the degree of exophthalmos and orbital pressure.Endoscopic transnasal inferomedial wall decompression can provide better improvement of visual function in patients with DON, but has a higher risk of surgery-related complications in comparison with the balanced orbital decompression.
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Objective:To measure the choroidal thickness (CT) in the macular area of patients with amblyopia in severe congenital ptosis and to compare the thickness with that of severe congenital ptosis without amblyopia.Methods:A cross-sectional study was performed.Total of 83 eyes of 60 severe congenital ptosis patients were enrolled from January to October in the year 2019 in the Shanxi Eye Hospital, including 29 patients 37 eyes with amblyopia and 36 patients 46 eyes without amblyopia.Thirty-seven eyes of 37 normal people were included in the control group.All subjects underwent mydriatic optometry and the spherical equivalent (SE) was calculated.IOL-Master was used to measure the axial length (AL), and EDI-OCT was used to measure the CT in the macular area.The CT was measured at 13 points: directly beneath the fovea (SFCT) and at 1 mm, 2 mm and 3 mm intervals to the nasal, temporal, superior and inferior to the fovea.The points of CT measurements were named according to their positions and distances relative to the fovea, and represented by SFCT, N1, N2, N3, T1, T2, T3, S1, S2, S3, I1, I2, I3 respectively.The SE, absolute astigmatism degree, AL and CT at each point were compared among the groups.The corrected AL covariance analysis was used to compare the CT at different measurement points among the groups.Spearman rank correlation analysis was used to evaluate the correlation between CT and AL or SE, and unitary linear regression analysis was used to evaluate the influence of age on AL in each group.This study was approved by the Ethics Committee of Shanxi Eye Hospital.All the subjects signed informed consent.Results:Compared with the non-amblyopia group and control group, the SE and absolute astigmatism value was larger, and the AL was shorter in the amblyopia group (all at P<0.05). The mean SFCT was (388.85±54.85), (301.48±56.68), and (287.08±61.88)μm in the amblyopia group, non-amblyopia group, and control group, respectively.Except for I3, the CT values of the rest parts in the amblyopia group were greater than those in the non-amblyopia group and the control group (all at P<0.05). Except for S3 and I3, the CT values of the rest parts in the amblyopia group were greater than those in the non-amblyopia group and the control group after adjusting the AL ( P<0.05). In all the three groups, the CT value was high in the subfovea and was thin in the nasal side.The SFCT was negatively correlated with AL (amblyopia group: rs=-0.340, P=0.045; non-amblyopia group: rs=-0.340, P=0.020; control group: rs=-0.463, P<0.001) and positively correlated with SE (amblyopia group: rs=0.350, P=0.039; non-amblyopia group: rs=0.412, P=0.004; control group: rs=0.450, P=0.005) in each group.Age was the influencing factor of AL in the non-amblyopia group and the control group ( β=0.243, 0.225; both at P<0.001). Conclusions:In comparison with severe congenital ptosis without amblyopia, the SE and absolute astigmatism degree are larger, the AL is shorter, and the CT values at most areas are higher in the severe congenital ptosis with amblyopia.CT value may be associated with the development of amblyopia in severe congenital ptosis.
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<p><b>OBJECTIVE</b>To determine the efficacy of endoscopic transnasal orbital balanced decompression technique for the treatment of severe Graves' orbitopathy.</p><p><b>METHODS</b>Six cases (12 eyes) of severe Graves'orbitopathy were included in this study. All patients were undergone endoscopic transnasal orbital balanced decompression technique (orbital inner and lower wall decompression+ lateral orbital decompression) for severe Graves'orbitopathy and serious affecting visual function or appearance.</p><p><b>RESULTS</b>The follow-up ranged from 12 months to 2 years and 4 months. Postoperatively, the visual acuity markly improved from 0.23 ± 0.08 to 0.48 ± 0.13 (t=-5.749, P<0.001). Proptosis measured by exophthalmoter distictly reduced from (22.08 ± 1.08) mm to (15.67 ± 1.44) mm (t=17.924, P< 0.001). The palpebral fissures obviously reduced from (12.03 ± 0.91) mm to (8.62 ± 0.75) mm (t=9.600, P< 0.001), and orbital volume evidently increased from (26.84 ± 0.62) cm³ to (31.56 ± 1.10) cm³ (t=-16.567, P<0.001). Compared with the preoperative result, the orbital pressure was significantly decreased after operation (t=-6.759 (Y=100 g), t=-11.850 (Y=200 g), t=-6.189 (Y=300 g), P<0.001). Diplopia was appeared in 3 cases, and disappeared 3 to 5 months after surgery.</p><p><b>CONCLUSION</b>Endoscopic transnasal orbital balanced decompression technique is effective for the treatment of severe Graves'orbitopathy.</p>
Subject(s)
Humans , Decompression, Surgical , Methods , Endoscopy , Exophthalmos , Pathology , Graves Ophthalmopathy , General Surgery , Orbit , General Surgery , Postoperative Period , Visual AcuityABSTRACT
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.
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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.
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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.
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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.