A long-term follow-up of peripapillary retinoschisis with optic disc hypoplasia.

Peripapillary and macular retinoschisis are usually associated with optic disc pits. We report a rare case of peripapillary retinoschisis with optic disc hypoplasia. A 59-year-old woman presented with asthenopia. Her best-corrected visual acuity was 20/20 OD. Ophthalmoscopy of the right eye revealed peripapillary retinoschisis, optic disc hypoplasia and dilated and tortuous radial peripapillary capillaries. There was no obvious optic disc pit or vitreous traction on optical coherence tomography (OCT) or fluorescein angiography. OCT showed retinoschisis around the optic disc, a thin sheet of fenestrated tissue on the optic disc and absence of serous retinal detachment. These findings had been almost the same at a previous visit to our hospital 17 years previously. Peripapillary retinoschisis may occur in patients with optic disc hypoplasia. We report a case in which visual acuity and symptoms did not change significantly after 17 years of follow-up.

Neuroprotective effects of granulocyte colony-stimulating factor on ischemia-reperfusion injury of the retina.

PURPOSE: It has been reported that granulocyte colony-stimulating factor (G-CSF) provides neuroprotection in models in which neuronal cell death is induced. This research was designed to investigate the effects of G-CSF on neurodegeneration of the inner retinal layer in a rat model of ischemic reperfusion (I/R) injury. MATERIALS AND METHODS: Retinal ischemia was induced by increasing the intraocular pressure to 110 mm Hg for 45 min in the left eyes of the rats. A sham operation was carried out on the right eyes. G-CSF (100 µg/kg/day in 0.3 ml saline) or the same volume of saline was intraperitoneally injected just before the operation and continued for 4 consecutive days (a total of 5 consecutive days). Morphological examinations, including the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, were performed 7 days after I/R induction. The expression of phosphorylated AKT in the retina was examined by Western blot analysis and immunohistochemistry. RESULTS: Cell loss in the ganglion cell layer was more significantly reduced in the I/R-induced eyes of the G-CSF-injected rats than in the I/R-induced eyes of the saline-injected rats (20.3 vs. 6.6%). The inner retinal thickness ratios, such as the inner plexiform layer to the inner limiting membrane/outer nuclear layer and the inner nuclear layer/outer nuclear layer, were significantly better preserved in the I/R-induced eyes of the G-CSF-injected rats than in the I/R-induced eyes of the saline-injected rats. TUNEL assays showed fewer apoptotic cells in the retinal sections of the I/R-induced eyes of the G-CSF-injected rats. The phosphorylation of AKT (p-AKT/AKT) was upregulated in the retinas of the I/R-induced eyes of the G-CSF-injected rats. CONCLUSION: Our results demonstrated that systemic injection of G-CSF can protect retinal ganglion cells and inner retinal layers from I/R injury. The effects could be associated with the activation of AKT.

Identification of ARIA regulating endothelial apoptosis and angiogenesis by modulating proteasomal degradation of cIAP-1 and cIAP-2.

Endothelial apoptosis is a pivotal process for angiogenesis during embryogenesis as well as postnatal life. By using a retrovirus-mediated signal sequence trap method, we identified a previously undescribed gene, termed ARIA (apoptosis regulator through modulating IAP expression), which regulates endothelial apoptosis and angiogenesis. ARIA was expressed in blood vessels during mouse embryogenesis, as well as in endothelial cells both in vitro and in vivo. ARIA is a unique protein with no homology to previously reported conserved domain structures. Knockdown of ARIA in HUVECs by using small interfering RNA significantly reduced endothelial apoptosis without affecting either cell migration or proliferation. ARIA knockdown significantly increased inhibitor of apoptosis (cIAP)-1 and cIAP-2 protein expression, although their mRNA expression was not changed. Simultaneous knockdown of cIAP-1 and cIAP-2 abolished the antiapoptotic effect of ARIA knockdown. Using yeast 2-hybrid screening, we identified the interaction of ARIA with 20S proteasome subunit alpha-7. Thereafter, we found that cIAP-1 and cIAP-2 were degraded by proteasomes in endothelial cells under normal condition. Overexpression of ARIA significantly reduced cIAP-1 expression, and this reduction was abolished by proteasomal inhibition in BAECs. Also, knockdown of ARIA demonstrated an effect similar to proteasomal inhibition with respect to not only expression but also subcellular localization of cIAP-1 and cIAP-2. In vivo angiogenesis studied by Matrigel-plug assay, mouse ischemic retinopathy model, and tumor xenograft model was significantly enhanced by ARIA knockdown. Together, our data indicate that ARIA is a unique factor regulating endothelial apoptosis, as well as angiogenesis, presumably through modulating proteasomal degradation of cIAP-1 and cIAP-2 in endothelial cells.

[Long-term surgical results of initial trabeculotomy combined with sinusotomy performed inferiorly].

PURPOSE: To evaluate retrospectively the long-term effects of initial trabeculotomy combined with sinusotomy performed inferiorly. PATIENTS AND METHOD: Enrolled were 128 eyes of 100 patients who received initial glaucoma surgery. In 36 eyes, the removal of Schlemm's canal endothelium was also performed (removed group). The results were compared with the intact group RESULTS: In the primary open angle glaucoma (POAG), mean intraocular pressure (IOP) at 3 years after surgery was 14.6 (intact) and 15.4 mmHg (removed). Kaplan-Meier life-table analysis showed that qualified success rates for the intact group at 8 years were 62.2% and for the removed group at 5 years 45.2% defined by 20 mmHg or lower. The results in developmental glaucoma (DG) were similar to those in POAG. No statistical differences in postoperative IOP between the intact and removed groups were seen in either POAG or DG. In exfoliation glaucoma (XFG), mean IOPs for the intact group at 3 years were 17.3 mmHg and for the removed group at 2 years 15.4 mmHg. The success rates for the intact group at 3.5 years were 25.2% and for the removed group at 4.5 years 64.3%. The results in the intact group were worse than in the POAG patients. Although visual disturbance was seen in 13% of the patients, the major cause was the progression of the cataracts. CONCLUSIONS: The long-term results were the same as those of previous reports on surgery performed superiorly, including the frequency of visual disturbance. However the removal of Schlemm's canal endothelium is necessary in XFG for better IOP control.

Administration of granulocyte colony-stimulating factor to recipients followed by intra-bone marrow-bone marrow transplantation accelerates acceptance of allogeneic bone marrow cells in mice.

We have recently established a novel method for bone marrow transplantation: intra-bone marrow-bone marrow transplantation (IBM-BMT), by which the rapid recovery of donor-derived hematopoiesis can be expected even when reduced radiation doses are used. In this paper, we examine, using mice, whether the combination of pretreatment of recipients with granulocyte-colony-stimulating factor (G-CSF) and IBM-BMT can induce a more rapid recovery of donor-derived hematopoiesis than IBM-BMT alone. We first pretreated recipients with recombinant human (rh) G-CSF (250 microg/kg/day) for 5 consecutive days (days -6 to -2). On day -1, the recipients were irradiated, and IBM-BMT was carried out on day 0. On day 12, we performed colony-forming units of spleen (CFU-S) assays. The combination of G-CSF pretreatment and IBM-BMT augmented the CFU-S counts, the weight of spleens, and the numbers of donor-derived hematopoietic cells. We next analyzed the mechanisms underlying these effects of G-CSF and found that (i) G-CSF induces Th2 polarization, which can prevent graft rejection, and (ii) G-CSF augments natural suppressor activity, which suppresses graft rejection. The combination of G-CSF pretreatment and IBM-BMT can produce the rapid recovery of donor-derived hematopoiesis and suppress graft rejection. This method would lighten the burden on patients in allogeneic BMT.

Posterior sub-Tenon injection of triamcinolone acetonide as pretreatment for focal laser photocoagulation in diabetic macular edema patients.

PURPOSE: To determine whether a posterior sub-Tenon injection of triamcinolone acetonide (TA) before focal photocoagulation is safe and effective in patients with diabetic macular edema. METHODS: Sixteen eyes of 11 diabetic patients with unresolved diffuse macular edema were treated with a 20-mg sub-Tenon injection of TA 1 to 2 months before focal photocoagulation. Focal photocoagulation was applied only to microaneurysms, and grid laser photocoagulation was not performed. The main outcome measures used were visual acuity (VA), central macular thickness (CMT) determined by optical coherence tomography (OCT), and the fluorescein angiographic appearance of the retina. Patients were followed for at least 6 months. RESULTS: One month after the sub-Tenon injection of TA, the macular edema was resolved with a significant reduction of the CMT on OCT. VA improved slightly. Subsequent focal photocoagulation of the microaneurysms maintained the significant reduction of CMT for up to 6 months. A significant improvement of VA was observed in 37.5% patients at 6 months, and there was no decrease in VA in any of the patients. CONCLUSIONS: A 20-mg sub-Tenon TA injection prior to focal laser photocoagulation is a safe and beneficial treatment in patients with diabetic macular edema.

Analyses of expression of cytoglobin by immunohistochemical studies in human tissues.

Cytoglobin (Cygb) is a recently discovered member of the vertebrate globin family, which includes probably most extensively studied proteins, hemoglobin (Hb), myoglobin (Mb) and neuroglobin (Ngb). It has been reported that Cygb is expressed ubiquitously at the mRNA or protein level. However, details of the distribution of Cygb in the various tissues have hitherto been unclear. In this experiment, we clarified the distribution of Cygb in various human tissues by immunohistochemical staining. First, we prepared a rabbit anti human Cygb polyclonal antibody. Using the antibody, we stained a tissue array slide containing 60 normal tissues from 40 human organs. We confirmed the staining patterns of the antibodies in these various tissues using autopsy samples from our university. In general, Cygb is positive in the epithelial cells, hepatocytes, pancreatic acinar cells, cardiomyocytes and skeletal muscle but rarely so in cells in the interstitial tissues. Cytoglobin is usually positive in the cytoplasm, but is also positive in the nucleus in some hepatocytes. In contrast, Cygb is negative in the smooth muscle. The distribution of Cygb could suggest its roles.

Reduction of Intraocular Pressure by Additional Trabeculotomy Ab Externo in Eyes With Primary Open-angle Glaucoma.

PURPOSE: The purpose of this study was to determine whether a second trabeculotomy (LOT) can reduce the intraocular pressure (IOP) in eyes with primary open-angle glaucoma (POAG) that had undergone an unsuccessful LOT as the initial surgery. PATIENTS AND METHODS: LOT ab externo was performed as a second surgery on 37 eyes of 34 POAG patients who had undergone an unsuccessful LOT as the initial surgery. The main outcome measure was the postoperative IOPs, and surgical failures were defined as eyes with a post-LOT IOP>20 mm Hg. The eyes were divided into 3 groups; those that underwent LOT as both the initial and additional surgery (L-L group), those that underwent LOT as the initial surgery and combined LOT and cataract surgery (cLOT-IOL) as the additional surgery (L-cL group), and those that underwent cLOT-IOL as the initial surgery and LOT as the additional surgery (cL-L group). RESULTS: The IOP was reduced after the additional LOT at postoperative 24 months in the L-L group from 20.0±3.0 mm Hg to 15.3±2.6 mm Hg (P<0.001), the L-cL group from 19.8±1.6 mm Hg to 15.8±3.2 mm Hg (P=0.029), and the cL-L group from 20.1±2.7 mm Hg to 15.5±2.3 mm Hg (P=0.014). There were no differences in the preoperative and postoperative IOPs between the initial-operated and additional-operated eyes. The success rates were improved by the additional surgery in the L-L group (P<0.001) and the L-cL group (P=0.029), but the rate was worsened in the cL-L group (P<0.001). CONCLUSIONS: These results indicate that LOT is a reasonable choice as an additional glaucoma surgery after failure of an initial LOT.

Plasma concentration of pigment epithelium-derived factor in patients with diabetic retinopathy.

CONTEXT: Pigment epithelium-derived factor (PEDF) is a strong inhibitor of angiogenesis. Eyes with diabetic retinopathy have low levels of ocular PEDF; however, the PEDF levels in the blood of diabetics have still not been determined. OBJECTIVES: Our objective was to determine the plasma levels of PEDF in diabetic patients and to determine the relationship with the stage of the diabetic retinopathy. DESIGN AND SETTING: This study was designed as a cross-sectional, institutional study. PATIENTS OR OTHER PARTICIPANTS: A total of 145 Japanese were studied; 112 had type 2 diabetes mellitus, and 33 were healthy controls. INTERVENTION: There was no intervention. MAIN OUTCOME MEASURES: The plasma level of PEDF was measured by ELISA, and the stage of diabetic retinopathy was determined by ophthalmic examinations. Clinical systemic status of diabetic patients was also examined. RESULTS: The plasma PEDF level in diabetic patients (6.68 +/- 0.54 microg/ml; mean +/- sem) was significantly higher than that in controls (4.38 +/- 0.59 microg/ml, P = 0.03), and the level was especially high in patients with proliferative diabetic retinopathy (7.78 +/- 0.98 microg/ml; n = 45; P = 0.005). The gender (P = 0.03), blood urea nitrogen (P = 0.005), and triglycerides (P = 0.04) were significant and independent determinants of plasma PEDF levels in diabetic patients. CONCLUSIONS: The PEDF level in the plasma was significantly elevated in diabetic patients, especially those with proliferative diabetic retinopathy. High levels of PEDF in the plasma may be related to the progression of diabetic retinopathy.

Leukostasis and pigment epithelium-derived factor in rat models of diabetic retinopathy.

PURPOSE: Spontaneously diabetic Torii (SDT) rats, an animal model of type 2 diabetes, have a low incidence of neovascular formation and an absence of non-perfused areas in their retinas at the proliferative stage that presents tractional retinal detachment with fibrous proliferation. The aim of this study was to determine whether leukostasis is present in the retina, to evaluate the levels of pigment epithelium-derived factor (PEDF) and intracellular adhesion molecule-1 (ICAM-1) levels in the blood of SDT rats, and to examine the effects of PEDF on leukostasis. METHODS: SDT rats, streptozotocin-induced diabetic (STZ) rats, and control Sprague-Dawley (SD) rats were studied. The index of leukostasis in the retina was determined immunohistochemically by counting the number of labeled adherent leukocytes. The levels of PEDF and the soluble intracellular adhesion molecule (sICAM)-1 in the plasma were measured. To investigate the effect of PEDF and vascular endothelial growth factor (VEGF) on leukostasis, the adhesion of monocytes to human umbilical vein endothelial cells (HUVECs) was assayed in vitro. RESULTS: SDT and STZ diabetic rats showed a significant increase of retinal leukostasis compared to that of control SD rats, but SDT rats had noteworthy lower levels of leukostasis than STZ rats in long term experiments. The sICAM-1 levels and PEDF expression were up-regulated in both STZ and SDT rats, but the SDT rats showed significantly higher levels of PEDF than STZ rats. In vitro studies showed that exposure of HUVECs to VEGF increased the number of adhering monocytes, and PEDF inhibited the VEGF-induced leukostasis in a dose-dependent manner. CONCLUSIONS: The inhibition of the VEGF-induced leukostasis by PEDF is most likely responsible for the low incidence of capillary occlusion and retinal neovascularization in SDT rats.

Immunotherapy for malignant tumors using combination of allogeneic intra-bone marrow-bone marrow transplantation, donor lymphocyte infusion and dendritic cells.

We have previously shown that the combination of allogeneic intra-bone marrow-bone marrow transplantation (IBM-BMT) and donor lymphocyte infusion (DLI) using CD4+ cell-depleted spleen cells is effective in suppressing tumor growth, but that this does not induce graft-versus-host disease (GVHD) in mice. In this report, we show that formalin-fixed tumor cell-pulsed dendritic cells (FFTCP DCs) have an additive effect with IBM-BMT plus DLI on the suppression of tumor growth, but that the DCs do not augment GVHD. BALB/c mice, which had been subcutaneously inoculated with Meth A (BALB/c-derived fibrosarcoma), were irradiated at a low dose (5 Gy) and were transplanted with bone marrow cells (BMCs) from C57BL/6 (B6) mice into the bone marrow cavity (IBM-BMT). Simultaneously, the mice were intravenously injected with spleen cells from B6 mice, and subcutaneously injected with FFTCP DCs derived from the bone marrow (BM) of B6 mice. At the point of the induction of DCs from BMCs, formalin-fixed Meth A cells were added into the culture. The mice treated with the combination of FFTCP DCs, IBM-BMT and DLI using CD4+ cell-depleted spleen cells showed smaller tumor sizes and longer survival than the mice treated with IBM-BMT plus FFTCP DCs or IBM-BMT plus DLI using CD4+ cell-depleted spleen cells. These results suggest that the combination of FFTCP DCs, IBM-BMT plus DLI using CD4+ cell-depleted spleen cells has potent anti-tumor effects without showing GVHD.

An innovative approach to bone marrow collection and transplantation in a patient with beta-thalassemia major: marrow collection using a perfusion method followed by intra-bone marrow injection of collected bone marrow cells.

Using small animals (mice and rats) and monkeys, we have found that the combination of bone marrow collection using the perfusion method (PM) and intra-bone marrow-bone marrow transplantation (IBM-BMT) of the collected cells is safe and effective in treating various intractable diseases. Based on these findings, we attempted to apply this method to humans. We report here the first case of a patient (6 years old) with beta-thalassemia major who underwent allogeneic BMT using this new PM + IBM-BMT method. The white blood cell counts of the patient gradually increased to more than 1500/microL by day 47 and continued to increase, reaching the highest level (8600/microL) on day +55. Fluorescence in situ hybridization data on day +33 showed that 98% of the peripheral blood cells were from the donor. Notably, there were no symptoms of graft-versus-host disease (GvHD). However, on day +56, the patient regrettably died of asphyxia resulting from sticky sputum. There was no evidence of infection (in the lung or liver) or GvHD (in the skin) by necropsy. We hope that this case report will help make our new strategies more readily available for the treatment of patients with various intractable diseases.

Human cord blood cells can differentiate into retinal nerve cells.

Retinal degeneration and dystrophy are the major causes of blindness in the developed world. It has been reported that human cord blood cells (HCBCs) can differentiate into neuron-like cells in vitro. We have recently demonstrated that bone marrow cells (BMCs) of both mice and rats can differentiate into retinal nerve cells (RNCs). In the present study, we show the differentiation capacity of HCBCs into RNCs in vivo. We transplanted lineage-negative HCBCs into the subretinal space of severe combined immunodeficiency (SCID) mice. Two weeks after the transplantation, some of the transplanted cells expressed human nestin, human MAP2, human neuron specific enolase (NSE), beta-III tubulin and also rhodopsin. These results indicate that HCBCs can differentiate into RNCs and suggest that our new strategy could be used for the regeneration of retinal nerve cells in degenerative or dystrophic diseases.

G-CSF and/or M-CSF accelerate differentiation of bone marrow cells into endothelial progenitor cells in vitro.

It has been reported that granulocyte-colony stimulating factor (G-CSF) and granulocyte-macrophage-colony stimulating factor (GM-CSF) can mobilize endothelial progenitor cells (EPCs) in bone marrow cells (BMCs) into peripheral blood (PB) in vivo. Previously, we also reported that macrophage-colony stimulating factor (M-CSF) can mobilize EPCs into PB, which results in the rapid recovery of blood flow in induced-ischemia limbs by augmenting the number of intramuscular capillaries in vivo. In the present study, we demonstrate that M-CSF and/or G-CSF can increase EPCs from lineage (CD3, B220, Gr-1, Mac-1, CD11c, Ter119, NK1.1 or CD31)-negative BMCs in vitro. Lineage-negative BMCs were cultured with or without M-CSF and/or G-CSF. Three days after culture with M-CSF and/or G-CSF, the number of Flk-1+/CD45-, Sca-1+/CD45-, CD31+/CD45- or CD146+/CD45- cells increased in comparison with no cytokines. When the cultured BMCs with or without G-CSF and/or M-CSF were intravenously injected into ischemia-induced hindlimbs of mice, the number of intramuscular capillaries in the ischemia-induced legs increased; BMCs cultured with G-CSF and/or M-CSF were more effective than those of cytokine non-treated BMCs. These results suggest that M-CSF and/or G-CSF can induce the differentiation of BMCs into EPCs, even in vitro.

Synergistic effects of granulocyte-colony stimulating factor and macrophage-colony stimulating factor on recovery of donor hematopoietic cells in allogeneic bone marrow transplantation.

G-CSF and M-CSF are used clinically to augment hematopoiesis after bone marrow transplantation (BMT) and chemotherapy. In this paper, we examined the synergistic effect of G-CSF and M-CSF on hematopoietic recovery in allogeneic BMT as a model of human BMT. We performed BMT from eGFP-transgenic mice (C57BL/6 background; H-2b) into lethally-irradiated C3H (H-2k). From the day after BMT, G-CSF and/or M-CSF were injected for 5 consecutive days. Not only the numbers of day 12 CFU-S and spleen weight, but also white blood cell (WBC) counts in the peripheral blood (PB) and nuclear cells in the bone marrow (BM) increased in the mice treated with G-CSF and/or M-CSF 12 days after BMT. Moreover, the number of donor-type WBCs in the PB and donor-type nuclear cells in the BM also increased in the mice treated with G-CSF and/or M-CSF. The effects were pronounced when G-CSF and M-CSF were used together rather than independently. These results suggest that treatment with the combination of G-CSF and M-CSF has a synergistic effect on hematopoiesis in allogeneic BMT.

Expression of pigment epithelium-derived factor and vascular endothelial growth factor in fibrovascular membranes from patients with proliferative diabetic retinopathy.

PURPOSE: The expression of pigment epithelium-derived factor (PEDF), a strong inhibitor of angiogenesis, has not been examined in human ocular fibrovascular membranes, to the best of our knowledge. The purpose of this study was to determine whether PEDF is expressed in the fibrovascular membranes in eyes of patients with proliferative diabetic retinopathy (PDR), and to compare the expression of PEDF with that of vascular endothelial growth factor (VEGF). METHODS: The expression of PEDF and VEGF in the fibrovascular membranes excised during vitreous surgery in eight cases of PDR was determined by immunohistochemistry. RESULTS: VEGF was strongly expressed in the endothelial cells of newly formed vessels in the fibrovascular membranes. In contrast, PEDF was weakly expressed in the endothelial cells and was prominently expressed in the extracellular matrix and fibrous tissue surrounding the new vessels. CONCLUSIONS: Our results suggest that PEDF, along with VEGF, may modulate the formation of fibrovascular membranes in patients with PDR.

Long-term survival of bone marrow-derived retinal nerve cells in the retina.

Recently, we have demonstrated that bone marrow stem cells can differentiate into retinal nerve cells. In the present study, we show a new and efficient strategy for transplanting bone marrow stem cells into the retina. When bone marrow stem cells were injected into the vitreous cavity of untreated eyes, only very few cells were found in the retina 2 weeks after injection. In contrast, when laser photocoagulation was performed just before the injection of bone marrow stem cells, a large number of the injected cells survived 2 weeks after injection and the cells expressed neural cell-specific or retinal nerve cell-specific antigens. Moreover, we still detected bone marrow stem cell-derived retinal nerve cells in the retina 1 year after injection in the retina.

Trans-Tenon's retrobulbar injection of triamcinolone acetonide for diffuse diabetic macular edema.

PURPOSE: To determine whether a trans-Tenon's retrobulbar injection of triamcinolone acetonide (TA) is a safe and effective treatment for diffuse diabetic macular edema. METHODS: Thirty-nine eyes of 30 diabetic patients with persistent macular edema were treated with 20 mg of TA injection. Central macular thickness (CMT) determined by optical coherence tomography (OCT) and visual acuity were evaluated before the injection and at 1, 2, 3, and 6 months, and up to 1 year in some eyes, after the injection. RESULTS: The CMT decreased significantly from 478 +/- 129 microm (mean +/- SD) before injection to 316 +/- 102 microm at 1 month, 307 +/- 104 microm at 2 months, and 275 +/- 89 microm at 3 months after a single injection of TA. A 20% reduction of CMT from the initial value was maintained by a single injection of TA in 27 of 39 eyes (69.2%) at 3 months, in 14 of 22 eyes (63.6%) at 6 months, and in 5 of 7 eyes at 12 months. A recurrence of macular edema was observed in 10% of the eyes at 3 months, and in 22.7% at 6 months. The 17 eyes in which vitrectomy had been carried out had a more significant improvement in CMT than the eyes without vitrectomy. CONCLUSION: A 20-mg trans-Tenon's retrobulbar TA injection is a safe and effective treatment for diabetic macular edema.

[Vitrectomy for proliferative diabetic retinopathy].

PURPOSE: We reviewed the outcome of vitrectomy for proliferative diabetic retinopathy (DR) and evaluated factors affecting the final visual outcome. METHODS: We performed primary vitreous surgery for proliferative DR in 148 eyes of 118 cases in three years from July 1999 to August 2002. All cases were followed for at least 3 months. We excluded vitreous surgery for diabetic maculopathy. Ages ranged from 24 to 80 (mean 57) years. Average postoperative follow-up period was 15 months. We evaluated the stage of DR by the new Fukuda classification. RESULT: Preoperative classification consisted of BIV (54 eyes, 36%), BV (94 eyes, 64%), and BV + VI (36 eyes). Final visual acuity was improved by 2 lines or more in 102 eyes (69%), remained unchanged in 28 eyes (19%), and decreased by two lines or more in 18 eyes (12%). There was a statistical correlation between preoperative visual acuity and final visual acuity. Earlier stages of DR had better visual outcome. Compared to the surgical outcome in the 1990s, the percentage of worsened eyes decreased. CONCLUSION: Vitrectomy for proliferative DR may be beneficial if performed in the earlier stages of DR or if the patient has better visual acuity before vitrectomy.

Immature dendritic cells (CD11c+ CD3- B220- cells) present in mouse peripheral blood.

It is well known that dendritic cells (DCs) are developed from the peripheral blood of mice when peripheral blood mononuclear cells (PBMCs) are cultured with GM-CSF. We have previously found that immature DCs are present in the blood even in humans. In the present study, we show that CD11c+ CD3- B220- cells in the mouse peripheral blood are immature DCs. The percentage of CD11c+ CD3- B220- cells in the (PBMCs) of normal mice ranges from 0.5 to 2.5%. The CD11c+ CD3- B220- cells in the PBMCs show dendrites, similar in shape to the CD11c+ CD3- B220- cells in the spleen, which are thought to be DCs definitely. However, they have practically no capacity to stimulate the proliferation of allogeneic T cells, and show a lower expression of MHC class II, B7-1 and B7-2 than CD11c+ CD3- B220- cells in the spleen. When the CD11c+ CD3- B220- cells in the PBMCs are cultured with GM-CSF, they show not only the potent ability to stimulate the proliferation of allogeneic T cells but also a higher expression of MHC class II, B7-1 and B7-2. Moreover, they migrate into the spleen when they are injected intravenously. These results suggest that CD11c+ CD3- B220- cells in the PBMCs are immature DCs, and that they migrate into the spleen, where they mature.