Objective To study the effects of several neurotrophic factors and growth factors on the survival of human retinal ganglion cells(RGC)in vitro. Methods RGC were isolated from donor eyes and cultured.RGC in cell culture were identified by morphologic criteria and immunocytochemical staining.Various neurotrophic factors and growth factors were added individually to the cultures.Numbers of RGC in wells in which these agents had been added were compared with those from control wells(cultures without supplements). Results No or very few RGC were present in cell cultures containing medium without supplements or those supplemented with neurotrophin-3(NT-3),nerve growth factor (NGF),epidermal growth factor(EGF)amd plateletderived growth factor(PDGF).Numbers of RGC(per 10 fields)in cell cultures containing brain derived neurotrophic factor(BDNF),ciliary neurotrophic factor(CNTF),neurotrophin-4/5(NT-4/5)and basic fibroblast growth factor(bFGF)wer 4.08,1.23,2.63 and 2.65,respectively,significantly more than found in the control cultures. Conclusions BDNF,NT-4/5,bFGF,CNTF improve survival of human RGC in vitro,while NGF,NT-3,EGF and PDGF do not. (Chin J Ocul Fundus Dis, 1999, 15: 149-152)
Objective To observe the incidence of ciliary detachment and its relationship with intraocular hypotension soon after vitrectomy. Methods A total of 46 eyes of 46 patients who underwent vitrectomy were examined by ultrasound biomicroscope (UBM). The patients were divided into three groups according to different tamponade: gas group (11 eyes), silicone oil group (8 eyes) and balanced saline solution (BSS) group (27 eyes). Basing on the postoperative intraocular pressure (IOP), the patients were divided into two groups: IOPlt; 10 mm Hg (25 eyes) and IOPge;10 mm Hg (21 eyes). UBM was applied to determine the tomographic features of the ciliary body 3 days after the surgery. IOP were monitored by noncontact tonometer daily after the surgery. The eyes with ciliary detachment were then examined once a week till the ciliary detachment reattached. The followup period was from 14 to 35 days. Results After vitrectomy, ciliary detachment was observed in 20 eyes of 46 eyes (43.5%), The incidence of ciliary detachment was 27.3％ in gas group, 25.0％ in silicone oil group, and 55.6％in BSS group. The average IOP in eyes with ciliary detachment ［(6.47plusmn;4.49) mm Hg （1 mm Hg=0.133 kPa）］ was significantly lower than that in the eyes without ciliary detachmen ［(15.61plusmn;7.72) mm Hg］ (t=8.031,Plt;0.001). The incidence of ciliary detachment was higher in eyes with postoperative IOP lt;10 mm Hg (68.0％) than that in the eyes with postoperative IOP ge;10 mm Hg (14.3%) (chi;2＝15.60, Plt;0.001). All eyes with postoperative ciliary detachment got spontaneous reattachment within 30 days after the surgery except that one eye had optic disc edema due to severe intraocular hypotension. Conclusions Early postoperative ciliary detachment is a common complication after vitrectomy. Eyes filled with BSS have the highest incidence of postoperative ciliary detachment. Most of the patients with ciliary detachment may get spontaneous reattahment within 30 days after the surgery.
ObjectiveTo observe the morphological and functional changes of retinal degeneration in mice with CLN7 neuronal ceroid-lipofuscinosis, and the therapeutic effects of glial cell derived neurotrophic factor (GDNF) and/or ciliary neurotrophic factor (CNTF) based on neural stem cells (NSC) on mouse photoreceptor cells. MethodsA total of 100 CLN7 mice aged 14 days were randomly divided into the experimental group and the control group, with 80 and 20 mice respectively. Twenty C57BL/6J mice aged 14 days were assigned as wild-type group (WT group). Mice in control group and WT group did not receive any interventions. At 2, 4, and 6 months of age, immunohistochemical staining was conducted to examine alterations in the distribution and quantity of cones, rod-bipolar cells, and cone-bipolar cells within the retinal of mice while electroretinography (ERG) examination was utilized to record scotopic a and b-waves and photopic b-wave amplitudes. At 14 days of age, the mice in the experimental group were intravitreally injected with 2 μl of CNTF-NSC, GDNF-NSC, and a 1:1 cell mixture of CNTF-NSC and GDNF-NSC (GDNF/CNTF-NSC). Those mice were then subdivided into the CNTF-NSC group, the GDNF-NSC group, and the GDNF/CNTF-NSC group accordingly. The contralateral eyes of the mice were injected with 2 μl of control NSC without neurotrophic factor (NTF) as their own control group. At 2 and 4 months of age, the rows of photoreceptor cells in mice was observed by immunohistochemical staining while ERG was performed to record amplitudes. At 4 months of age, the differentiation of grafted NSC and the expression of NTF were observed. Statistical comparisons between the groups were performed using a two-way ANOVA. ResultsCompared with WT group, the density of cones in the peripheral region of the control group at 2, 4 and 6 months of age (F=285.10), rod-bipolar cell density in central and peripheral retina (F=823.20, 346.20), cone-bipolar cell density (F=356.30, 210.60) and the scotopic amplitude of a and b waves (F=1 911.00, 387.10) in central and peripheral retina were significantly decreased, with statistical significance (P＜0.05). At the age of 4 and 6 months, the density of retinal cone cells (F=127.30) and b-wave photopic amplitude (F=51.13) in the control group were significantly decreased, and the difference was statistically significant (P＜0.05). Immunofluorescence microscopy showed that the NSC transplanted in the experimental group preferentially differentiated into astrocytes, and stably expressed CNTF and GDNF at high levels. Comparison of retinal photoreceptor nucleus lines in different treatment subgroups of the experimental group at different ages: CNTF-NSC group, at 2 months of age: the whole, central and peripheral regions were significantly different (F=31.73, 75.06, 75.06; P＜0.05); 4 months of age: The difference between the whole area and the peripheral region was statistically significant (F=12.27, 12.27; P＜0.05). GDNF/CNTF-NSC group, 2 and 4 months of age: the whole (F=27.26, 27.26) and the peripheral area (F=16.01, 13.55) were significantly different (P＜0.05). In GDNF-NSC group, there was no statistical significance at all in the whole, central and peripheral areas at different months of age (F=0.00, 0.01, 0.02; P＞0.05). ConclusionsCLN7 neuronal ceroid-lipofuscinosis mice exhibit progressively increasing degenerative alterations in photoreceptor cells and bipolar cells with age growing, aligning with both morphological and functional observations. Intravitreal administration of stem cell-based CNTF as well as GDNF/CNTF show therapeutic potential in rescuing photoreceptor cells. Nevertheless, the combined application of GDNF/CNTF-NSC do not demonstrate the anticipated synergistic protective effect. GDNF has no therapeutic effect on the retinal morphology and function in CLN7 neuronal ceroid-lipofuscinosis mice.
Objective To investigate the mRNA expression of ciliary neurotrophic factor on the retina during injury and repair of optic nerves in rats. Methods Thirty-five healthy SD rats were randomly divided into 3 groups: 5 in the control group, 15 in the simply transected optic nerve group and 15 in the optic nerve-sciatic nerve anastomosis group. The simply transected and optic nerve-sciatic nerve anastomosed models were set up, and the retinal tissues of all of the rats were taken out after 3, 7 and 14 days, respectively; and the mRNA expression of CNTF in the 3 groups were observed by semiquantitative reversal transcription-polymerase chain reaction method. Results A minimum expression of CNTF mRNA was found in the retinae of the control group, and the increased rates of expression were found in the retinae of the simple transection of optic nerve group with the increase rate of 100%, 594%, and 485% on the 3rd, 7th, and 14th day respectively after the operation, while in optic nerve-sciatic nerve anastomosis group, the increase rates were found to be 258%, 752% and 515% on the 3rd, 7th, and 14th day respectively after the operation. Conclusion Retinal neurons can respond to axonal reaction of retinal ganglion cells by up-regulate endogenous CNTF after the injury of the optic nerves, which may provide a theoretic base for the application of the exogenous CNTF. (Chin J Ocul Fundus Dis，2004，20：355-357)
Ciliary body tumor is a rare intraocular tumor. Due to its unique anatomical location, its correct diagnosis and reasonable treatment are very difficult problems. In terms of diagnosis and differential diagnosis, ophthalmologists need to fully utilize the role of slit lamp microscope and transillumination experiment to capture secondary changes in the anterior segment caused by hidden ciliary body tumors, such as monocular localized cataract, lens indentation, and pigment dissemination, etc. Ophthalmological imaging methods, especially ultrasound biomicroscopy, can achieve the purpose of early detection and early diagnosis. According to the size, location and morphological characteristics of the tumor, a reasonable treatment plan is formulated. Since ciliary body tumors are mostly benign, the recurrence rate of local resection is low, which can satisfy the pathological diagnosis and preserve part of the patient's vision. Therefore, eye-preserving treatment should be advocated. However, enucleation remains the treatment of choice for tumors that are too large to be treated with local excision or radiation, eyes with refractory glaucoma, and tumors that do not respond to radiation therapy.
ObjectiveTo analyze the ultrasonographic features of adenoma of the nonpigmented ciliary epithelium (ANPCE). MethodsA retrospective series of case studies. From January 2014 to October 2021, 31 patients (31 eyes) with ANPCE (ANPCE group) were diagnosed in the eye center of Beijing Tongren Eye Center of Beijing Tongren Hospital, Capital Medical University, and 17 patients (17 eyes) with ciliary body melanoma (control group) diagnosed at the same time were selected as the control group. There was no significant difference in age (t=－0.564) and sex composition ratio (χ2=0.182) between the two groups (P=0.576, 0.670). All patients underwent ultrasound biomicroscopy to obtain the measurement parameters: tumor height, maximum basal diameter, maximum diameter, ratio of maximum diameter to basal diameter and ratio of maximum diameter to height; tumor location, shape, internal echogenicity intensity, echogenicity uniformity, degree of sound attenuation, invasion of iris, anterior displacement of the iris, lens subluxation were observed. The measurement parameters and observation indexes of the two groups were compared by independent sample t-test and χ2 test. Receiver operating characteristic (ROC) curve was drawn, area under the ROC curve (AUC) was determined, and parameter indicators with differential diagnosis value were screened. ResultsThe maximum diameter, height, maximum basal diameter, ratio of the maximum diameter to the maximum basal diameter, and the ratio of the maximum diameter to the height of the tumors in the ANPCE group and the control group were 5.64±0.98 mm, 4.24±0.59 mm, 3.66±0.71 mm, 1.58±0.34, 1.34±0.19 and 7.82±2.03 mm, 4.47±2.44 mm, 7.02±1.96 mm, 1.13±0.16, 2.09±1.06. The maximum diameter, the maximum basal diameter, and the ratio of the maximum diameter to the height of the tumor in the ANPCE group were all smaller than those of the control group, and the ratio of the maximum diameter to the maximum basal diameter was greater than that of the control group, and the differences were statistically significant (t=－4.159, －6.808,－2.924, 6.257; P<0.05). The tumors in the ANPCE group were mainly spherical (87.1%, 27/31), with no significant acoustic attenuation (77.4%, 24/31), less invading the root iris (77.4%, 24/31), and the tumors were mostly located in the ciliary body coronal (74.2%, 23/31); tumors in the control group were mainly hemispherical (47.1%, 8/17) or spherical (47.1%, 8/17), with significant sound attenuation (76.5%, 13/17), most of the tumors invaded the iris (70.6%, 12/17), and the tumors were mostly located from the pars plana to the coronal (76.5%, 13/17). There were statistically significant differences in the position, shape, sound attenuation degree, and whether it invaded the iris between the two groups of eyes (χ2=15.132, 19.767, 13.118, 10.581; P<0.05). The results of ROC curve analysis showed that the ratio of the largest diameter to the largest base diameter, the degree of sound attenuation and the AUC of whether the iris was violated were higher, which were 0.881, 0.769, and 0.740, respectively. ConclusionsUltrasound biomicroscopy is helpful in the diagnosis and differential diagnosis of ANPCE and ciliary body melanoma. The ratio of maximum diameter to maximum basal diameter, the degree of sound attenuation and whether it invades the root iris are important parameters to distinguish the two tumors.
Objective To probe the significance of application of ultrasound biomicroscopy (UBM) in the diagnosis and management of the iris and ciliary tumors. Methods UBM (Mode 840, Humphrey, 50 MHz 5 mm×5 mm) was done in 34 cases (35 eyes) of iris and ciliary body tumors, and some of the affected eyes underwent B-scan or Doppler ultrasound and CT scan. Histopathological examination of the resected tumor tissues was performed in 21 eyes of the operation. Results Among this series of 35 eyes with iris and ciliary body tumors detected by UBM, the characteristics of locality and solidity of the tumors, i,e., anterior chamber in plantation cyst, cyst behind the iris, and solid tumors of iris and ciliary body, of 21 eyes undergone surgical treatment revealed the same results both in UBM and histopathological examinations. Conclusion UBM can supply precise informations in diagnosis and treatment of tumors of iris and ciliary body. (Chin J Ocul Fundus Dis, 2002, 18: 128-130)
Objective To investigate the effects of ambroxol hydrochloride on surface structure of trachea mucosa in rats injured by intratracheally instilled amikacin. Methods Thirty Wistar rats injured by intratracheally instilled amikacin ( 0. 252 mL/kg) were randomly divided into a control group ( n =15) and an ambroxol group ( n= 15) . The rats in the ambroxol group were intraperitoneally injected with ambroxol hydrochloride ( 70 mg/kg) 5 minutes after amikacin administration. They were all equally divided into five subgroups and sacrificed at 2, 4, 8, 28, 48 hours respectively. Then the samples of 1/3 lower segment of trachea were collected and observed under scanning electron microscope. Results In the control group, the mucous secretion and its stickness were increased. The cilia were found lodged, sticked together, aligned abnormally, abrupt partly, and recovered slowly, with the percentage of damaged area of 98. 2% , 98. 5% , 97. 5%, 92. 7% , 82. 1% at 2, 4, 8,24,48 h, respectively. The injuries of mucosa in the ambroxol group were much milder and recovered more rapidly than those in the control group, with the percentage of damaged area of 85. 7% , 81. 9% , 73. 0% , 61. 9% , 50. 2% at 2, 4, 8, 24, 48 h, respectively. Conclusions Intratracheal instillation of amikacin can cause cilia ultrastructure damage on tracheal mucosa. Ambroxol can promote the recovery process and alleviate airway inflammation.
Objective To establish a culture system in vitro of fetal and adult human retinal neural cells provide a model for the basic research of retinal neural cells and the medicinal exploitation. Methods Fetal human retinas(10~13 weeks after conception) and adult human retinas(20~40 years old) were dissected, dissociated, and put into culture plate which was coated with polylysine or rat tail gel. Specific growth factor EGF、FGF、BDNF or NT-4 were added to the culture medium. BrdU incorporation, Tunnel assessment and immuno-histochemistry and immuno-fluorescent staining were applied to determine cells proliferation, apoptosis and identify the component of cultured cells. Results Fetal human retinal cells and adult human retinal cells survived for up to 100 and 180 days in vitro. The addition of EGF、FGF、BDNF or NT-4 promoted the survival of both fetal and adult retinal neurons and stimultated proliferation of fetal retinal cells. The neurons or the rate of ganglion cells was observed with higher percentage in the group with growth factor adding than the group without. Conclusion Fetal and adult human retinal cells can be maintained in vitro and the fetal cells also can be expanded, which are helpful to generate retinal neurons for basic research and drug exploitation. The exogenous growth factors added to the culture medium can promote survival, proliferation and differentiation of retinal cells in culture. (Chin J Ocul Fundus Dis, 2002, 18: 279-282)