Effect of optokinetic stimulation on weight-bearing shift in standing and sitting positions in stroke patients.

BACKGROUND: Patients with hemiplegia after stroke tend to bear weight on the non-paretic side and exhibit large postural sway during static standing and walking, which may increase their risk of falls. Improvement of the sitting posture balance in the early phase of rehabilitation by adjusting weight-bearing would minimize the risk of falls as early rehabilitation reportedly improves walking ability and prevents falls in later phases of rehabilitation or at discharge. AIM: This study aimed to evaluate the effect of optokinetic stimulation (OKS) on shift of the weight-bearing (displacement of the center of pressure [CoP]) in patients with hemiplegia who are incapable of independent standing. DESIGN: Quasi-experimental, cross-sectional study. SETTING: Rehabilitation hospital. POPULATION: Patients with hemiplegia in the subacute phase after stroke (N.=37). METHODS: Standing and sitting balance tests were performed during OKS projected onto a screen. For OKS, a pattern of random dots was presented, which continuously moved in horizontal or torsional directions during both static standing and sitting conditions. Postural sway was assessed during standing and sitting by measuring the sway path, sway area, sway velocity, and mean displacement of CoP. The magnitude of the lateral change in CoP as an indicator of the weight-bearing shift was evaluated by subtraction of the mean CoP of the right-left axis component in the stationary condition from the mean CoP sway during OKS. RESULTS: OKS induced a unilateral change of the mean CoP position in patients during both, sitting and static standing, indicating that OKS can shift the weight-bearing in patients after stroke, irrespective of the posture condition. Moreover, the same OKS approach evoked an analogous shift in patients with more severe symptoms, with impairment in independent standing. CONCLUSIONS: OKS could induce a significant shift in weight balance in patients with hemiplegia after stroke who are incapable of independent standing, suggesting that the OKS approach can be applied to a broader spectrum of patients, including those with more severe symptoms. CLINICAL REHABILITATION IMPACT: OKS approach would improve exercise training in the early phase of rehabilitation of patients with hemiplegia after stroke.

Quality of Life and Utility Values for Cost-Effectiveness Modeling in Japanese Patients with Type 2 Diabetes.

INTRODUCTION: Reliable quality of life (QoL) measures and utility values are needed for patients with type 2 diabetes mellitus (T2DM) with a variety of comorbid conditions to help facilitate cost-effectiveness modeling. This study aimed to evaluate the Diabetes Treatment-Related Quality of Life (DTR-QOL) and EuroQol 5-dimension 5-level (EQ-5D-5L) questionnaires in patients with T2DM with and without diabetes complications and comorbidities in Japan. METHODS: This was an observational survey study involving 1000 patients with T2DM, at least 20 years old, receiving treatment at Nara University Hospital or Takamura Internal Medicine Clinic in Japan. Patients completed the DTR-QOL and EQ-5D-5L questionnaires and clinicians completed an accompanying case report form. The DTR-QOL and EQ-5D-5L are scored on a scale of 0-100 and 0-1, with 100 and 1 representing the best possible scores, respectively. RESULTS: Out of 1000 recruited patients, 978 were included in the final analysis. Patients reported an average EQ-5D-5L value of 0.92 ± 0.11. Utility values corresponded to the degree of severity of health conditions while few differences were observed when stratified by the HbA1c 7% threshold, age, or BMI level, nor did the values correspond to the degree of clinical risk factors. Patients reported an average total DTR-QOL score of 79.26 ± 13.26. The DTR-QOL was sensitive to detect differences in patients with T2DM with a variety of complications and comorbidities, risk factors, and treatments. CONCLUSION: This is the largest study to report QOL values for patients with diabetes in Japan and the first to include a variety of comorbid diabetic conditions. These findings may be useful for cost-effectiveness modeling of patients with T2DM in Japan.

Bone-Marrow-Derived Mononuclear Cells Relieve Neuropathic Pain after Spinal Nerve Injury in Mice.

Treating neuropathic pain is a critical clinical issue. Although numerous therapies have been proposed, effective treatments have not been established. Therefore, safe and feasible treatment methods are urgently needed. In this study, we investigated the therapeutic effects of autologous intrathecal administration of bone-marrow-derived mononuclear cells (MNCs) on neuropathic pain. We generated a mouse model of neuropathic pain by transecting the spinal nerve and evaluated neuropathic pain by measuring the mechanical threshold in the following 14 days. Mice in the MNC injection group had a higher mechanical threshold than those in the buffer group. We assessed the effect of MNC treatment on the dorsal root ganglia and spinal cord by immunohistochemistry, mRNA expression, and cytokine assay. The migration and accumulation of microglia were significantly suppressed in the MNC group, and the mRNA expression of inflammatory cytokines such as interleukin (IL)-6, IL-1ß, and tumor necrosis factor alpha (TNF-α) was markedly downregulated. Furthermore, MNC administration tended to suppress various cytokines in the cerebrospinal fluid of the model mice. In conclusion, our results suggest that intrathecal injection of MNCs relieves neuropathic pain and might be a promising cell therapy for the treatment of this condition.

Current status of achieving blood pressure target and its clinical correlates in Japanese type 2 diabetes.

AIMS/INTRODUCTION: To investigate the current status of achieved blood pressure levels in association with the number of antihypertensive drug classes as of 2013, and to explore the clinical correlates with achievement of target blood pressure in a large-scale cohort of Japanese subjects with type 2 diabetes. MATERIALS AND METHODS: A nationwide survey was conducted including 12,811 subjects with type 2 diabetes. Subjects were divided by achieved blood pressure, <130/80 or 140/90 mmHg, and the number of drug classes taken. RESULTS: The percentages achieving a blood pressure of <130/80 or 140/90 mmHg were 52.0% and 86.1%, respectively. The prevalence of hypertension, if defined as ≥130/80 mmHg or treated, became 67.9%. Among subjects taking antihypertensive drugs, a blood pressure of <130/80 or <140/90 mmHg was 46.7% and 83.2%, respectively. The percentages of <130/80 mmHg were 55.9% without drugs, 47.1% on 1, 42.5% on 2, 47.2% on 3, and 56.8% on ≥4 drugs, respectively. The most prescribed drugs were renin-angiotensin system inhibitors, followed by calcium channel blockers, diuretics, and ß-blockers. The multiple logistic regression analysis indicated that a blood pressure <130/80 mmHg was associated with lower values in age, body mass index, albuminuria, and glomerular filtration rate, higher proportions on targets for HbA1C and lipids, and less retinopathy. CONCLUSIONS: In type 2 diabetes, hypertension is common and only 52% achieved <130/80 mmHg, indicating a difficulty in blood pressure lowering. This was correlated with difficulties in glycemic and lipid management, obesity, and vascular complications, implying these clustering to be a serious problem. This article is protected by copyright. All rights reserved.

Large-scale survey of rates of achieving targets for blood glucose, blood pressure, and lipids and prevalence of complications in type 2 diabetes (JDDM 40).

OBJECTIVE: The fact that population with type 2 diabetes mellitus and bodyweight of patients are increasing but diabetes care is improving makes it important to explore the up-to-date rates of achieving treatment targets and prevalence of complications. We investigated the prevalence of microvascular/macrovascular complications and rates of achieving treatment targets through a large-scale multicenter-based cohort. RESEARCH DESIGN AND METHODS: A cross-sectional nationwide survey was performed on 9956 subjects with type 2 diabetes mellitus who consecutively attended primary care clinics. The prevalence of nephropathy, retinopathy, neuropathy, and macrovascular complications and rates of achieving targets of glycated hemoglobin (HbA1c) <7.0%, blood pressure <130/80 mm Hg, and lipids of low-density/high-density lipoprotein cholesterol <3.1/≥1.0 mmol/L and non-high-density lipoprotein cholesterol <3.8 mmol/L were investigated. RESULTS: The rates of achieving targets for HbA1c, blood pressure, and lipids were 52.9%, 46.8% and 65.5%, respectively. The prevalence of microvascular complications was ∼28% each, 6.4% of which had all microvascular complications, while that of macrovascular complications was 12.6%. With an increasing duration of diabetes, the rate of achieving target HbA1c decreased and the prevalence of each complication increased despite increased use of diabetes medication. The prevalence of each complication decreased according to the number achieving the 3 treatment targets and was lower in subjects without macrovascular complications than those with. Adjustments for considerable covariates exhibited that each complication was closely inter-related, and the achievement of each target was significantly associated with being free of each complication. CONCLUSIONS: Almost half of the subjects examined did not meet the recommended targets. The risk of each complication was significantly affected by 1 on-target treatment (inversely) and the concomitance of another complication (directly). Total diabetes care including one-by-one management of modifiable risk factors and complications may be important for high-quality care. The future studies including more subjects and clinics with precise complication status are needed.

Relationship between the efficacy of oral antidiabetic drugs and clinical features in type 2 diabetic patients (JDDM38).

AIMS/INTRODUCTION: We carried out an observational cohort study to examine the relationship between the efficacy of oral antidiabetic drugs and clinical features in type 2 diabetics. MATERIALS AND METHODS: We analyzed the CoDiC(®) database of the Japan Diabetes Data Management Study Group across 67 institutions in Japan. In a total of 3,698 drug-naïve patients who were initiated with metformin, dipeptidyl peptidase-4 inhibitor (DPP-4i) or sulfonylurea (SU) from 2007 to 2012, we evaluated body mass index (BMI) and hemoglobin A1c (HbA1c). The patients were stratified according to their clinical features, and matched using a propensity score to adjust for baseline factors. RESULTS: HbA1c was reduced with all drugs, with the largest effect elicited by DPP-4i and the smallest by SU (P = 0.00). HbA1c increased with SU after 6 months in the patients stratified by an age-of-onset of <50 years (P = 0.00). BMI increased with SU in the patients stratified by a BMI of <25 (P = 0.00), and decreased with metformin in the patients with a BMI >25 (P = 0.00). The reduction in HbA1c was larger in patients with HbA1c of ≥8%, compared with that in patients with HbA1c of <8% (P = 0.00). HbA1c during the study period was higher in patients who were added to or swapped with other drug(s), than in patients continued on the original drug (P = 0.00). CONCLUSIONS: The effect on bodyweight and glycemic control differed among metformin, DPP-4i and SU, and the difference was associated with clinical features.

Liraglutide Versus Sitagliptin in a 24-week, Multicenter, Open-label, Randomized, Parallel-group Study in Japanese Type 2 Diabetes Mellitus Patients Responding Inadequately to a Sulfonylurea and/or One or Two Other Oral Antidiabetic Drugs (JDDM 33).

OBJECTIVE: Liraglutide (glucagon-like peptide-1 [GLP-1] receptor agonist) and sitagliptin (dipeptidyl peptidase-4 inhibitor) are approved in Japan for treating type 2 diabetes mellitus (T2DM). We compared the efficacy and safety of adding liraglutide or sitagliptin to a sulfonylurea in Japanese T2DM patients. METHODS: Patients aged 18 to <80 years with hemoglobin A1c (HbA1c; National Glycohemoglobin Standardization Program [NGSP]) of 6.9-9.4%, body mass index ≤35 kg/m(2), and treatment with a sulfonylurea and/or one or two non-sulfonylurea oral antidiabetic drugs for greater than or equal to eight weeks before enrollment were eligible. Patients were randomized in an open-label manner to either 0.9 mg/day liraglutide (n = 50) or 50-100 mg/day sitagliptin (n = 49) and were treated for 24 weeks. Non-sulfonylureas were discontinued before randomization. Patients using other oral antidiabetic drugs started sulfonylurea treatment. The primary endpoint was the change in HbA1c from baseline to Week 24. RESULTS: HbA1c decreased in both groups, and the reduction was significantly greater throughout in the liraglutide group except for Week 24 (0.59 ± 0.80 vs. 0.24 ± 0.94%; P = 0.0525). Fasting plasma glucose (FPG) decreased significantly in the liraglutide group compared with the sitagliptin group (-21.15 ± 31.22 vs. +0.46 ± 39.39 mg/dL; P = 0.0014). Homeostasis model assessment of ß cell function and C-peptide increased significantly in the liraglutide group but not in the sitagliptin group. Hypoglycemic symptoms and adverse events occurred in four and nine patients, respectively, in the liraglutide group, and in two and five patients, respectively, in the sitagliptin group. CONCLUSION: Treatment with liraglutide or sitagliptin together with a sulfonylurea improved HbA1c in Japanese T2DM patients in primary care. Both drugs were associated with low rates of adverse events and hypoglycemia. The improvement in ß cell function probably contributed to the improvement in glycemic control in the liraglutide group.

Hyaluronan production and hyaluronan synthase expression in three human conjunctiva-derived cell strains.

PURPOSE: The conjunctiva maintains the health of the ocular surface by protecting the eye from pathogen invasion, injury, and dryness. In this study, we investigated the regulation of hyaluronan (HA) synthesis by cytokines in conjunctiva-derived cells. METHODS: Cultured primary cells derived from human conjunctivas that had been removed as surgical specimens were transfected with an immortalizing gene (human papilloma virus 16 E6/E7). To compare the biological features of the primary and immortalized cells, we assessed their morphological features and gene expression patterns. We also examined the effects of inflammatory cytokines on hyaluronan synthase (HAS) expression and HA production. RESULTS: Three conjunctiva-derived cell strains were established and could be passaged up to 15 times. All strains expressed ß2MG and KRT13 transcripts, highly expressed in conjunctival epithelial cells. HA production and expression of the three HAS isoforms were detected in the cell strains; however, cytokine treatment had no significant effect on HA production or HAS isoform expression. CONCLUSIONS: We succeeded in deriving three human cell strains from conjunctival tissue. In the conjunctiva-derived cell strains, HA production and HAS mRNA expression were stable and were not changed by either TGF-ß or PDGF-BB.

Cloning and characterization of human vitreous tissue-derived cells.

PURPOSE: Previously, we established a porcine vitreous tissue-derived hyalocyte cell line (PH5) and investigated the regulation of hyaluronan synthesis in these cells by cytokines. The objective of the current study was to establish human vitreous tissue-derived cells and to compare their characteristics with those of PH5 cells. METHODS: Human vitreous specimens from two patients were cultured in the presence of 10% foetal bovine serum and immortalized by infection with human papilloma virus 16 genes E6 and E7. We used reverse transcription polymerase chain reaction (RT-PCR) to analyse and compare the expression profiles for several genes in the human vitreous tissue-derived cells and PH5 cells. To investigate the regulation of hyaluronan production in response to cytokine stimulation, the expression of hyaluronan synthase isoforms was examined using RT-PCR, and hyaluronan production was measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: Two types of cells, HV64 and HV65, were derived from human vitreous tissue. The HV64 and HV65 cell-doubling times were 58 r and 76 hr, respectively. The cells expressed messenger RNA (mRNAs) encoding collagen type I α1 (COL1A1), collagen type II α1 (COL2A1), CD11b, CD14, CD68, CD204 and CD206 but did not express mRNA for glial fibrillary acidic protein (GFAP). Cytokine stimulation did not induce the expression of hyaluronan synthase mRNA or the production of hyaluronan. In contrast, mRNAs for GFAP and hyaluronan synthase-2 were expressed in the porcine PH5 cells, and treatment with transforming growth factor-ß1 and/or platelet-derived growth factor-BB induced the production of hyaluronan in PH5 cells. CONCLUSION: The new human vitreous tissue-derived cells have macrophage-like characteristics and are different from our previously developed porcine hyalocyte cells. These human vitreous tissue-derived cells might be useful for studies of human intraocular diseases.

Time and Costs of Insulin Treatment in the Care of Newly Registered Type 2 Diabetes Patients in Diabetes Clinics Across Japan (JDDM 22).

AIMS: To study the time and costs of insulin treatment of newly registered outpatients with Type 2 diabetes mellitus (T2DM). METHODS: In total, 355 patients with T2DM were registered on their first visit to one of 11 diabetes clinics across Japan. Of these, 313 were not being treated with insulin (the non-insulin group), whereas 42 were (the insulin group). In the insulin group, 26 were already on insulin at the first visit, whereas 16 were started on insulin after their first visit. The time and costs involved in the care were recorded over the following 5 months. RESULTS: In the first 3 months, considerable time was expended in both groups, with the time spent by physicians a little (but significantly) longer for the insulin group. The total time expended by all care providers was approximately 1.3-fold greater for the insulin compared with the non-insulin group. The total cost and total cost/min for the insulin group was almost twice that for the non-insulin group. Over the 5-month period, mean HbA1c in the non-insulin group improved from 8.0% to 6.5%, with 72% achieving a glycemic target of HbA1c ≤ 6.5%. In contrast, in the insulin group, mean HbA1c improved from 9.4% to 7.6%, with only 39% achieving the target. There were no reports of major hypoglycemic events in either group and body mass index remained stable. CONCLUSIONS: The insulin therapy for T2DM can be achieved safely and effectively at outpatient clinics, even though it requires considerably more time and resources than non-insulin therapy.

Cotylenin A inhibits cell proliferation and induces apoptosis and PAX6 mRNA transcripts in retinoblastoma cell lines.

PURPOSE: Retinoblastoma, a childhood cancer of the retina, is caused by inactivation of the tumor suppressor gene retinoblastoma (RB). Cotylenin A (CN-A), a novel fusicoccane-diterpene glycoside, accelerates the differentiation of several types of myeloid cell lines and is a candidate for a new type of anticancer therapeutic agent with this effect. However, whether CN-A has the same effect on retinoblastoma cells is unknown. We studied the response of two retinoblastoma cell lines, Y-79 and WERI-Rb-1, to CN-A. METHODS: We studied the response of two retinoblastoma cell lines to CN-A with respect to cell growth, apoptosis, morphology, mRNA, protein expression analysis of specific genes (N-myc, cyclin-dependent kinase inhibitor 1A [P21], paired box gene 6 [PAX6], and rhodopsin [RHO]), and activity of three PAX6 promoters (P0, P1, and Palpha). RESULTS: CN-A inhibited cell proliferation and induced apoptosis via caspase activity in the two retinoblastoma cell lines. In addition, CN-A induced mRNA expression of P21, PAX6, and RHO and protein expression of P21. In Y-79 cells, PAX6 P1 promoter was activated by CN-A. In WERI-Rb-1 cells, PAX6 P0, P1, and Palpha promoter were activated by CN-A. CN-A decreased mRNA and protein expression of N-myc in two retinoblastoma cell lines. CONCLUSIONS: The responses of retinoblastoma cells to CN-A include inhibition of cell growth, induction of apoptosis, and the potential to change neuroblastoma characteristics of retinoblastoma cells.

Association of PDGF-BB-induced thrombomodulin with the regulation of inflammation in the corneal and scleral stroma.

PURPOSE: The responses of corneal and scleral stromal cells to platelet-derived growth factor (PDGF)-BB were assessed and inflammatory reactions in the cornea and sclera were investigated. METHODS: Primary cultures of cells obtained from human subjects and strains derived from human corneal or scleral stromal cells (Cs3 and Sc1, respectively) were used. Changes in gene expression after 24 hours of exposure to 10 ng/mL PDGF-BB were analyzed with an Sc1 DNA microarray. The upregulation of several genes in Cs3 and Sc1 was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. The expression of bioactive factors was detected immunohistochemically in nine different clinical specimens. RESULTS: DNA microarray analysis revealed that the gene encoding thrombomodulin (TM) was induced in Sc1 by PDGF-BB. RT-PCR confirmed that TM expression at the mRNA level was increased in both corneal and scleral stromal cells. At the protein level, TM expression was increased in scleral stromal cells, but not in corneal cells, and TM was detected in both the membrane and cytoplasmic compartments. TM was detected immunohistochemically in inflamed scleral and several corneal specimens. After TM stimulation, interleukin (IL)-18 transcription was increased in Sc1. CONCLUSIONS: PDGF-BB induced TM mRNA expression in scleral and corneal stromal cells, but Western blot analysis revealed the increase in TM expression only in the scleral cells. TM induced IL-18 in scleral stromal cells. A cascade involving these biologically active factors may regulate scleral and corneal inflammation. The results also reveal differences in the biological response of scleral and corneal stromal cells.

Cloning and characterization of cell strains derived from human corneal stroma and sclera.

PURPOSE: To establish human corneal stroma- and sclera-derived cells as models for studying diseases of the anterior segment of the eye. METHODS: Using a recombinant retrovirus system, we transfected human papilloma virus 16 E6 and E7 (HPV16 E6/E7) into human corneal stroma- and sclera-derived cells. The primary cells and established cell strains were characterized by assessing the mRNA expression of collagen, matrix metalloproteinase, and tissue inhibitor of metalloproteinase by reverse transcription-polymerase chain reaction. We also examined the effects of inflammatory cytokines on hyaluronan synthase expression and hyaluronan products. RESULTS: Both a corneal stroma-derived cell strain, Cs3, and a sclera-derived cell strain, Sc1, were obtained, and both cell strains could be passaged up to 25 times. The mRNA expression pattern observed in the primary cells was identical to that observed in the cell strains. Hyaluronan synthase 1 and 2 mRNAs were increased by transforming growth factor beta and platelet-derived growth factor BB. Significant differences were observed between the hyaluronan products with and without cytokine treatment. CONCLUSION: Cell strains derived from corneal stroma and sclera fibroblast cells can be established using HPV16 E6/E7 immortalized genes of the same origin. The phenotypic cell characteristics did not change after transfection, immortalization, or successive passages in culture.

[A case of primary orbital adenocarcinoma responding to chemoradiation].

Primary orbital adenocarcinoma is very rare. There are not any reports about the treatment of this disease, except for surgery. We experienced a case of primary orbital adenocarcinoma, which we successfully treated by chemoradiation using 5-FU and cisplatin. It is very important to collect and record cases of rare diseases responding to certain chemotherapy regimens.

Clinicopathological analysis of malignant eyelid tumor cases at Yamagata university hospital: statistical comparison of tumor incidence in Japan and in other countries.

PURPOSE: To report the clinical and histopathological features of malignant eyelid tumor cases treated in our clinic. We also compared the differences in the frequency of malignant eyelid tumor in various regions of Japan and worldwide. METHODS: Retrospectively, we studied the records of the 38 cases of malignant eyelid tumor treated in Yamagata University Hospital over the last 17 years. The statistical comparison with various countries was based on reports of case studies in those countries. RESULTS: Data from our clinic: Among the total of 38 cases, 15 cases (39.5%) were diagnosed as basal cell carcinoma, 11 cases (28.9%) as sebaceous gland carcinoma, and 4 cases (10.5%) as squamous cell carcinoma. In addition, three cases were malignant melanoma, two adenocarcinoma, one Merkel cell carcinoma, one malignant peripheral nerve sheath tumor, and one malignant lymphoma. The ages of patients ranged from 45 to 92 years (mean, 72.0 +/- 12.4 years). Most of the cases were treated by complete resection of the tumors and eyelid reconstruction. Radiation or cryotherapy were added when required. The prognosis of the cases with basal cell carcinoma and squamous cell carcinoma was good, and that of the other tumors was relatively poor. During the same period, in Caucasians, basal cell carcinoma constituted about 80%-90% of the malignant eyelid tumors, whereas in Japan and Asian countries, basal cell carcinoma, sebaceous gland carcinoma, and squamous cell carcinoma each constituted about 20%-40%. CONCLUSIONS: A racial difference in the incidence of basal cell carcinoma, sebaceous gland carcinoma, and squamous cell carcinoma can be considered in making a diagnosis.

Immunohistochemical diagnosis of a rare case of epithelioid malignant peripheral nerve sheath tumor with multiple metastases.

BACKGROUND: The epithelioid variant of malignant peripheral nerve sheath tumor (MPNST) is a rare tumor with poor prognosis that sometimes involves the head and neck. The diagnosis is based principally on the histological examination, and it is generally very difficult to reach the correct diagnosis. CASE: An 84-year-old Japanese woman presented with a tumor mass of 2 week's duration in the right medial canthal region. OBSERVATIONS: Although the tumor was excised surgically, metastases occurred three times on her face and head, and the patient died of distant systemic multiple metastases. In the histopathological analysis, the tumor showed a composite pattern comprising spindle or polygonal cells arranged in irregular bands, and a population of larger epithelioid cells in solid sheets and nests. In the immunohistochemical analysis, the tumor cells were positive for S-100 protein, vimentin, and nerve growth factor receptor (NGFR), and negative for cytokeratin and HMB 45 (melanoma-associated antigen). These findings confirmed the diagnosis of MPNST. CONCLUSIONS: Epithelioid MPNST has complex histopathological findings and histopathological features similar to other epithelioid tumors, especially malignant melanoma. Immunohistochemical examination using NGFR and HMB-45 is important in the differential diagnosis.

Proteome analysis of human vitreous proteins.

PURPOSE: Various protein contents such as enzymes, growth factors, and structural components are responsible for biological activities in organs. We have created a map of vitreous proteins and developed a proteome analysis of human vitreous samples to understand the underlying molecular mechanism and to provide clues to new therapeutic approaches in eyes with proliferative diabetic retinopathy (PDR). METHODS: Vitreous and serum samples were obtained from subjects with idiopathic macular hole (MH, 26 cases) and PDR (33 cases). The expressed proteins in the samples were separated by two-dimensional (2-D) polyacrylamide gel electrophoresis. Protein spots were visualized by silver staining, and their expression patterns were analyzed. Some protein spots of concern were excised from the 2-D gels, digested in situ with trypsin, and analyzed by mass spectrometry. RESULTS: More than 400 spots were detected on 2-D gels of MH cases, of which 78 spots were successfully analyzed. The spots corresponded to peptide fragments of 18 proteins, including pigment epithelium-derived factor, prostaglandin-D2 synthase, and interphotoreceptor retinoid-binding protein. These were not identified in the corresponding serum samples. These proteins were also expressed in PDR samples, with no distinct tendency to increase or decrease compared with the MH samples. More than 600 spots were detected on 2-D gels of PDR cases, of which 141 spots were successfully analyzed. The spots corresponded to peptide fragments of 38 proteins. Enolase and catalase were identified among four detected spots. Neither was found in MH vitreous or in PDR serum samples. CONCLUSION: A map of protein expression was made in human vitreous from eyes with MH and PDR. In the PDR eyes, the increased protein expression observed was due to barrier dysfunction and/or production in the eye. Proteome analysis was useful in systematic screening of various protein expression in human vitreous samples.