Integrin-Linked Kinase Controls Choroidal Neovascularization by Recruitment of Endothelial Progenitor Cells.

Purpose: Vasculogenesis has been shown to contribute to the formation of choroidal neovascularization (CNV). However, the mechanism behind the recruitment of endothelial progenitor cells (EPC) to CNV is not well understood. Therefore, we were interested to know whether integrin-linked kinase (ILK) plays a role in recruiting EPC to CNV, and its possible mechanism. Methods: We investigated the effect of hypoxia on retinal pigment epithelium (RPE) cells expressing ILK, hypoxia-inducible factor 1α (HIF-1α), stromal-derived factor-1 (SDF-1), and vascular endothelial growth factor (VEGF), and we further examined the effect of ILK small interfering RNA (siRNA) on their expression. The function of ILK expressed by RPE on EPC in vitro with regard to angiogenic effect was also studied. In vivo, we determined the expression levels of the above factors in CNV. We also examined the role of ILK on their expression, on EPC recruiting, and on the growth of CNV. Results: We found that hypoxia strongly induced the expression of ILK, HIF-1α, SDF-1, and VEGF. Moreover, the silencing of ILK attenuated their expression. It also decreased the phosphorylation of protein kinase B (PKB/AKT) and extracellular regulated protein kinases (ERK) and nearly abolished the proliferation, migration, and adhesion of EPC to RPE cells. In vivo, we showed that these factors were upregulated in CNV. Inhibiting the expression of ILK prohibited the "homing" of EPC to CNV lesions and attenuated the growth of CNV. Conclusions: We demonstrate that ILK controls the development of CNV by regulating the recruitment of EPC to CNV lesions, possibly through ILK-dependent expression of SDF-1 and VEGF in RPE.

Neurological complications of thoracic posterior vertebral column resection for severe congenital spinal deformities.

PURPOSE: The risk of neurological injury during vertebral column resection is high. In this study, we investigated the incidence and risk factors for neurological complications when treating spinal deformities by thoracic posterior vertebral column resection (PVCR). METHODS: Between 2008 and 2013, there were 62 consecutive patients (34 female patients and 28 male; the mean age: 16.3 years, range 6-46 years) treated with thoracic PVCR. We retrospectively reviewed the clinical records to obtain demographic and radiographic data, operative time, estimated blood loss (EBL, the ratio between circulating and lost blood), bleeding volume (the lost blood), number of vertebrae fused, number of vertebrae resected, usage of titanium mesh cage, and intraoperative neuromonitoring data. Multi-factor logistic regression was used to find the major risk factors for neurological complications. RESULTS: The average follow-up period was 46 months (range 24-88 months); no patients were lost to follow up. The average operative time was 524.8 ± 156.8 min (range 165.0-880.0 min), the average bleeding volume was 2585 ± 2210 ml (100-9600 ml), and the average estimated blood loss was 75.8% (9-278%). Ten patients (16.1%) developed post-operative neurological complications (nine transient and one permanent). Multi-factor logistic regression revealed that the risk factors for neurological complications were age ≥18 years, pulmonary dysfunction, and EBL >50%. CONCLUSIONS: Thoracic PVCR can lead to satisfactory outcomes in the treatment of severe spinal deformities. Risk factors for neurological complications include the age over 18 years, presence of pulmonary dysfunction, and EBL greater than 50%. The pulmonary dysfunction can be regarded as the most valuable indicator to measure the severity of the spine deformity.

Safety and Efficacy of Single-Stage Surgical Treatment for Congenital Scoliosis Associated with Intraspinal Mass.

For congenital scoliosis associated with intraspinal anomaly, surgical treatment is often advocated. However, the safety and efficacy of single-stage intraspinal mass resection and scoliosis correction remain unclear. The purpose of this study was to retrospectively evaluate the feasibility and risk factors of single-stage surgical treatment for congenital scoliosis associated with intraspinal mass. Patients' clinical records were reviewed for demographic and radiographic data, operating time, intraoperative blood loss, perioperative complications, and postoperative pathologic results. Two female and 5 male patients with an average age of 19.14 ± 7.52 years (range, 11-31 years) were evaluated. Patients were followed for a minimum of 24 months after initial surgical treatment, with an average of 49.71 ± 32.90 months (range, 27-99 months). Spinal curvature was corrected from an average of 69.57 ± 20.44° to 29.14 ± 9.87°, demonstrating a mean correction rate of 55.05% ± 18.75%. No obvious loss of correction was observed at the final follow-up. Complications included transient neurologic deficit, cerebrospinal fluid leakage, and intraspinal mass recurrence in 1 patient each. There was no paralysis or permanent nerve damage. In conclusion, simultaneous intraspinal mass resection and scoliosis correction appears to be safe and effective.

Expression of soluble Fas and soluble FasL in human nucleus pulposus cells.

The study aimed for addressing the expression of soluble Fas (sFas) and soluble Fas Ligand (sFasL) in human nucleus pulposus (NP) and its attendant relationship with disc degeneration. Human NP samples were collected from patients with disc degeneration and cadavers as degenerate and normal groups, respectively. Subsequently, NP cells were cultured in monolayer. ELISA was performed to identify the expression levels of sFas and sFasL in the supernatant of NP cell cultures in vitro. Quantitative real-time PCR was used to detect the expression of sFas and sFasL in human NP cells in mRNA solution. The study comprised 12 degenerate and 8 normal cadaveric NP samples. The concentration value of sFas in the supernatant was significantly higher from degenerate NP than that from normal NP at each time point. In contrast, sFasL was significantly lower at each time point. Moreover, the expression of sFas and sFasL reached the peak at various early stages of cell cultures and decreased thereafter. Furthermore, the mRNA level of Fas in degenerate NP cells was significantly higher than that in normal cells; whereas FasL showed an opposite pattern. The study is the first addressing the expression of sFas and sFasL in human NP cell cultures. Moreover, the expression of sFas and sFasL varies with culture time in vitro with different levels in degenerate and normal settings. These findings indicate that sFas and sFasL might play a role in intervertebral disc degeneration.

[Possible mechanism of endothelial progenitor cells in the development of rat choroidal neovascularization].

OBJECTIVE: To investigate the role and possible mechanism of endothelial progenitor cell (EPC) in the development of choroidal neovascularization (CNV). METHODS: Experimental study. Twenty-four BN rats were divided into 3 groups.One eye of each animal was induced by laser photocoagulation with 532 nm laser and the contralateral eye was taken as control. Three, seven and fourteen days after photocoagulation the formation of CNV was observed by histopathological study and the recruitment of EPC and the possible pro-angiogenic growth factors released by EPC during the development of CNV were examined by multi-labeled immunofluorescence staining. The difference among the 3 groups was analyzed by ANOVA and the comparison between any 2 groups was further checked by LSD-t test. RESULTS: Both the histopathological study and the immunofluorescence staining indicated that within the laser lesions proliferated and migrated cells grew into the subretinal space through the broken Bruch membrane 3 days after photocoagulation, 7 days after photocoagulation lumen-like structures were observed and CNV became stable until 14 days after photocoagulation. No EPC was observed in the normal retina whereas EPC were recruited into the laser lesions 3 days after photocoagulation, comprising (79.29 ± 11.27)% of the total endothelial cell population within CNV. At 7-day EPC constituted new vessels within CNV area and the proportion decreased to (47.13 ± 5.78)%. Then its number decreased dramatically 14 days after photocoagulation contributing to (10.83 ± 2.79)% of the endothelial cells in CNV. The differences either among the 3 groups (F = 104.623, P < 0.05) or between any 2 groups (P < 0.05) were statistically significant. Moreover, triple labelled immunofluorescence staining showed that the EPC within CNV area could also secret pro-angiogenic factors such as vascular endothelial growth factor, IL-10, bFGF and MMP9. CONCLUSION: EPC involves in the development of CNV not only through participating in the formation of new vessels within the CNV area but also through secreting pro-angiogenic factors.

[Expression of Hsp70 Grp94 and IgG in human lung carcinoma].

AIM: To explore the expression and significance of heat shock protein 70 (Hsp70), glucose regulated protein 94 (Grp94) and immunoglobulin G (IgG) in human lung carcinoma. METHODS: The expression of Hsp70, Grp94 and IgG in 40 human lung carcinomas was studied using immunohistochemical technique and image analysis. The localization among Hsp70, Grp94 and IgG was analyzed by double labeling immunofluorescent staining and laser scanning confocal microscopy. RESULTS: Hsp70, Grp94 and IgG in Human lung carcinomas showed high expression. The positive rate of Hsp70, Grp94 and IgG was 65% (26/40), 45% (18/40), and 82.5% (33/40), respectively. The average value of optical density was 5.10 +/- 0.32, 3.52 +/- 0.35, and 8.12 +/- 0.31, respectively. Hsp70 was localized in nucleus and cellular cytoplasm while Grp94 and IgG were mainly localized in cellular cytoplasm. Ten cases showed Hsp70 was co-localized with IgG and Eighteen cases showed Grp94 was co-localized with IgG. CONCLUSION: High expression of Hsp70, Grp94 and IgG in Human lung carcinomas suggested that Hsp70, Grp94 and IgG might play an important role in the development of human lung carcinoma. IgG is co-localized with Hsp70 or Grp94. The study will lay a theorical basis for further research on anti-tumor immunotherapy.