Ferroptosis-Related lncRNA to Predict the Clinical Outcomes and Molecular Characteristics of Kidney Renal Papillary Cell Carcinoma.

Kidney renal papillary cell carcinoma (KIRP) is a highly heterogeneous type of kidney cancer, resulting in limited effective prognostic targets for KIRP patients. Long non-coding RNAs (lncRNAs) have emerged as crucial regulators in the regulation of ferroptosis and iron metabolism, making them potential targets for the treatment and prognosis of KIRP. In this study, we constructed a ferroptosis-related lncRNA risk score model (FRM) based on the TCGA-KIRP dataset, which represents a novel subtype of KIRP not previously reported. The model demonstrated promising diagnostic accuracy and holds potential for clinical translation. We observed significant differences in metabolic activities, immune microenvironment, mutation landscape, ferroptosis sensitivity, and drug sensitivity between different risk groups. The high-risk groups exhibit significantly higher fractions of cancer-associated fibroblasts (CAFs), hematopoietic stem cells (HSC), and pericytes. Drugs (IC50) analysis provided a range of medication options based on different FRM typing. Additionally, we employed single-cell transcriptomics to further analyze the impact of immune invasion on the occurrence and development of KIRP. Overall, we have developed an accurate prognostic model based on the expression patterns of ferroptosis-related lncRNAs for KIRP. This model has the potential to contribute to the evaluation of patient prognosis, molecular characteristics, and treatment modalities, and can be further translated into clinical applications.

A pilot study of viscoelastic agent to prevent recurrent vitreous hemorrhage after vitrectomy for proliferative diabetic retinopathy.

BACKGROUND: To evaluate the possibilty of preventing recurrent vitreous hemorrhage (RVH) after vitrectomy in proliferative diabetic retinopathy (PDR) patients with unabsorbed vitreous hemorrhage (VH) by intravitreal injection of viscoelastic agent (VA) at the end of the surgery and compared its effect with triamcinolone acetonide (TA). METHODS: This was a pilot prospective, observational study. PDR patients with VH who underwent vitrectomy were assigned to 3 groups according to the tamponade applicated at the end of the surgery, including VA group (intravitreally injected 1 ml VA if the retina was prone to bleed during the operation), TA group (intravitreally injected 2 mg TA when there was much exudates), or balanced salt solution (BSS) group (no tamponade). Then postoperative follow-up was performed routinely until 6 months after surgery. The primary outcome was the incidence of RVH, secondary outcome were the best-corrected visual acuity (BCVA) and introcular pressure (IOP). Cataract formation and other complication were also assessed. RESULTS: A total of 68 eyes, from 68 patients, were included. 18,18,32 eyes were enrolled in the VA group, TA group and BSS group, respectively. The integral incidence of RVH after vitrectomy was 5.6%, 5.6% and 12.5% respectively (P = 0.602). There was no early RVH in VA or TA group, whereas 3 early RVHs were identified in BSS group, however there was no significant difference (P = 0.171). Every group had one late RVH case. In all groups, final BCVA showed significant improvement compared to baseline. BCVA at any postoperative visit showed no significant differences among 3 groups. Mean IOP was higher 1 week after surgery in VA group compared with the other groups; however, in other times the differences were not significant. No cataract formation and other complication was noted in 3 groups. CONCLUSION: Intravitreal injection of VA or TA at the end of vitrectomy for PDR patients with unabsorbed VH tend to reduce the incidence of early RVH after vitrectomy similarly. As VA was preferred to applicate in the eyes that were prone to bleed, intravitreal injection of VA at the end of vitrectomy might be a promising method for preventing RVH in PDR patients.

Acupuncture treatment of facial spasm: An overview of systematic reviews.

BACKGROUND: To provide evidence, this review evaluated the methodological quality, risk of bias, and reporting quality of SRs/MAs in the treatment of Facial Spasm with acupuncture. METHODS: Systematic reviews and Meta analyses (SRs/MAs) of acupuncture for Facial Spasm were retrieved from 8 databases from inception to October 1, 2022. Two reviewers independently screened the literature and extracted the data, then used Assessment of Multiple Systematic Reviews-2 (AMSTAR-2), Bias Risk in Systematic Review (ROBIS), and Preferred Report Item for Systematic review and Meta-analysis (PRISMA), Grading of Recommendations, Assessment, Development and Evaluation (GRADE) to assess methodological quality, risk of bias, quality of reporting, and quality of evidence. RESULTS: A total of 8 SRs/meta-analyses were included. All the SRs were published between 2012-2022. Based on AMSTAR-2, 8 SRs were rated critically low quality. By using the ROBIS tool, 6 SRs were rated low-risk bias. With the PRISMA-A checklist, we found 2 out of 8 SRs were found adequately reported over 70%. With the GRADE system, no high-quality evidence was found, and only two outcomes provided moderate-quality evidence. Among the downgraded factors, the risk of bias within the original trials was ranked first, followed by publication bias, inconsistency, and imprecision. CONCLUSION: Acupuncture is a promising complementary treatment for HFS. However, due to the low quality of the SRs/MAs supporting these results, high-quality studies with rigorous study designs and larger samples are needed before widespread recommendations can be made.

Effect of central serous chorioretinopathy patients by covering sick eye for 48 hours / Efeito da cobertura ocular por 48 horas de pacientes com coriorretinopatia serosa central

ABSTRACT Purpose: To assess the effect of continuously covering the sick eye affected with central serous chorioretinopathy for 48 h. Methods: This retrospective, case-control study involved 32 central serous chorioretinopathy patients categorized in the treatment group composed of 17 sick eye that received continuous covering treatment for 48 h with a medical gauze and the observation group composed of 15 of these patients who were followed up. None of the patients received any other treatments or medications. The logarithm of the minimal angle of resolution (logMAR) best-corrected visual acuity, macular retinal thickness, and the root mean square value of the amplitude density in the first ring of multifocal electroretinogram were examined before and after the 48-h treatment. Results: After the continuous treatment, the logMAR best-corrected visual acuities were 0.31 ± 0.18 and 0.56 ± 0.37 in the treatment and observation groups, respectively (p=0.019). The macular retinal thicknesses were 461 ± 43 µm and 498 ± 50 µm in the treatment and observation groups, respectively (p=0.032). The root mean square values of the amplitude density in the first ring of multifocal electroretinogram were 32.5 ± 5.3 nV/deg2 and 26.6 ± 4.3 nV/deg2 in the treatment and observation groups, respectively (p=0.002). Conclusions: The continuous application of the covering treatment for 48 h on the sick eye showed positive outcomes with respect to the best-corrected visual acuity, macular retinal thickness, and macular retina functions in the treatment of central serous chorioretinopathy.

RESUMO Objetivo: Este estudo teve como objetivo avaliar o efeito da cobertura contínua do olho doente de pacientes com coriorretinopatia serosa central por 48 horas. Métodos: Este estudo retrospectivo, caso-controle, incluiu 32 pacientes com coriorretinopatia serosa central, dos quais 17 receberam tratamento de cobertura contínua por 48 horas no olho doente com gaze médica como grupo de tratamento e 15 foram acompanhados como grupo de observação. Todos os pacientes não receberam nenhum outro tratamento ou medicamento. O logaritmo do ângulo mínimo de resolução da acuidade visual melhor corrigida (LogMar), a espessura macular da retina e o valor médio da raiz quadrada da densidade da amplitude no primeiro anel do eletroretinograma multifocal foram examinados antes e após o tratamento por 48 horas, respectivamente. Resultados: Após o tratamento contínuo, a acuidade visual melhor corrigida pela escala logMar foi de 0, 31 ± 0, 18 no grupo de tratamento e 0, 56 ± 0, 37 no grupo de observação (p=0, 019). A espessura macular da retina foi de 461 ± 43 µm no grupo de tratamento e 498 ± 50 µm no grupo de observação (p=0,032). O valor médio da raiz quadrada da densidade de amplitude no primeiro anel do eletroretinograma multifocal foi de 32,5 ± 5,3 nV/deg2 no grupo com cobertura e foi de 26,6 ± 4,3 NV/deg2 no grupo de observação (p=0,002). Conclusões: O tratamento de cobertura contínua no olho doente, durante 48 horas, apresentou efeitos positivos na acuidade visual melhor corrigida, na espessura e na função macular da retina no tratamento da coriorretinopatia serosa central.

Effect of central serous chorioretinopathy patients by covering sick eye for 48 hours.

PURPOSE: To assess the effect of continuously covering the sick eye affected with central serous chorioretinopathy for 48 h. Methods: This retrospective, case-control study involved 32 central serous chorioretinopathy patients categorized in the treatment group composed of 17 sick eye that received continuous covering treatment for 48 h with a medical gauze and the observation group composed of 15 of these patients who were followed up. None of the patients received any other treatments or medications. The logarithm of the minimal angle of resolution (logMAR) best-corrected visual acuity, macular retinal thickness, and the root mean square value of the amplitude density in the first ring of multifocal electroretinogram were examined before and after the 48-h treatment. RESULTS: After the continuous treatment, the logMAR best-corrected visual acuities were 0.31 ± 0.18 and 0.56 ± 0.37 in the treatment and observation groups, respectively (p=0.019). The macular retinal thicknesses were 461 ± 43 µm and 498 ± 50 µm in the treatment and observation groups, respectively (p=0.032). The root mean square values of the amplitude density in the first ring of multifocal electroretinogram were 32.5 ± 5.3 nV/deg2 and 26.6 ± 4.3 nV/deg2 in the treatment and observation groups, respectively (p=0.002). CONCLUSIONS: The continuous application of the covering treatment for 48 h on the sick eye showed positive outcomes with respect to the best-corrected visual acuity, macular retinal thickness, and macular retina functions in the treatment of central serous chorioretinopathy.

Ultrastructural study of closed macular hole- preliminary application of a novel high magnification module combining with OCT.

BACKGROUND: As a novel high magnification module (HMM) combining with OCT (OCT-HMM) is able to detect the microstructure of retina, we apply it to explore the ultrastructure of the macula after closure of the idiopathic macular hole (IMH) by surgery. METHODS: This is an observational case series study in which patients with full-thickness IMHs who had undergone successful macular closure by vitrectomy and internal limiting membrane peeling and healthy subjects were recruited. After comprehensive ophthalmic examinations, the images of macular area were obtained and collected by professional operators using OCT-HMM. Then images were independently analyzed by 4 masked vitreoretinal specialists. RESULTS: A total of 24 IMH eyes and 42 healthy eyes were examined. HMM images were obtained in 10 IMH eyes. Among them, 4 eyes whose macula closed completely with recovery of photoreceptor layer presented a dark arc nasal to the fovea, oriented to the optic, and the notch of arc faced temporally. Six eyes in which the macula closed incompletely with photoreceptor cells loss revealed a dark ring with uneven bright spots inside. The other 14 eyes failed to obtain clear images by OCT-HMM. The contra lateral eyes of the patients and the healthy subjects' eyes succeeded to obtain the HMM images which displayed evenly grey background thickly covered with tiny bright dots that was in similar size and evenly and widely distributed and there no dark arc or ring. OCT B-scan and IR images could be acquired in all of the IMH and healthy eyes. CONCLUSION: The preliminary application of HMM has supplied us a brand-new insight into the microstructure of closed IMH. A dark arc sign could be detected with OCT-HMM in the macula which was functionally closed after surgery that was probably the healing mark on a microstructure photoreceptors level. Its existence and shape indicated that the functional closure followed by a retinal displacement mainly horizontally from temporal side to nasal side but not symmetric centripetally.

Effects of Simulated Microgravity on Ultrastructure and Apoptosis of Choroidal Vascular Endothelial Cells.

BACKGROUND: It was confirmed that simulated microgravity (SMG) led to ultrastructural alterations and apoptosis in many types of microvascular endothelial cells. However, whether SMG would also affect choroidal vascular endothelial cells (CVECs) remains unknown. This study was designed to investigate the effects of SMG on ultrastructure and apoptosis of CVECs. METHODS: The rotary cell culture system (RCCS) was utilized to simulate microgravity condition. Human CVECs were cultured under normal gravity (NG) or SMG condition for 3 days. The ultrastructure was viewed under transmission electron microscopy, and the organization of F-actin was observed by immunofluorescence staining. Additionally, the apoptosis percentage was calculated using flow cytometry. Moreover, the mRNA and protein expression of BAX, Bcl-2, Caspase3, Cytochrome C, p-AKT, and p-PI3K were detected with quantitative PCR and Western blot at different exposure time. RESULTS: In the SMG group, CVECs presented with a shrunk cell body, chromatin condensation and margination, mitochondria vacuolization, and apoptotic bodies. The amount of F-actin decreased, and the filaments of F-actin were sparse or even partly discontinuous after cultivation under SMG for 72 h. The proportions of apoptotic CVECs in SMG groups at 24 and 72 h were significantly higher than those in the NG group (P < 0.001). The mRNA and protein expression of Bax, Caspase3, and Cytochrome C of CVECs in SMG groups at 24 and 72 h significantly increased than those of the NG group, respectively (P < 0.001). The alterations of p-AKT and p-PI3K protein expression possessed similar trends. On the contrary, the mRNA and protein expression of Bcl-2 in CVECs under SMG at 24 and 72 h were significantly less than that of the NG group, respectively (P < 0.001). CONCLUSION: Simulated microgravity conditions can lead the alterations of the F-actin structure and apoptosis of CVECs. The Bcl-2 apoptosis pathway and PI3K/AKT pathway may participate in the damage of CVECs caused by SMG.

Effects of Foxp3 gene modified dendritic cells on mouse corneal allograft rejection.

OBJECTIVE: To investigate the effect of Foxp3 gene modified dendritic cells (Foxp3 + DC) on allogeneic T cells proliferation and to study the effect of Foxp3 + DC on corneal allograft rejection. METHODS: Lentivirus-Foxp3 was transfected into DC2.4 cells, as Foxp3 + DC cells. 42 BALB/c mice were randomly divided into: Group A (n = 6), normal group; Group B (n = 12), Group C (n = 12) and Group D (n = 12), allograft groups, were treated with normal saline, DC2.4, Foxp3 + DC by intraperitoneal injection, respectively. RESULTS: Compared with the control group, Foxp3 protein in the Foxp3 + DC cells increased significantly (P < 0.05); the expressions of CD80 and CD86 immunophenotypes of Foxp3 + DC cells decreased significantly (P < 0.05); IL-12 secretion reduced (P < 0.05), but IL-10 secretion was promoted (P < 0.05). The average transplant survival time in Group B was (14.833 ± 1.472) d, and Group C and Group D led to a statistically significant prolongation of transplant survival to (17.667 ± 1.366, 23.000 ± 2.000) d (P < 0.05) respectively. 14 d after transplantation, as compared with Group C and D, the expressions of IFN-γ in grafts markedly increased in Group B. 14 d after transplantation, as compared with Group B, the expressions of Foxp3 mRNA, IDO mRNA in grafts decreased remarkably in Group C and D (P < 0.05); as compared with Group C, the expressions of Foxp3 mRNA, IDO mRNA in grafts decreased remarkably in Group D (P < 0.05). CONCLUSION: Foxp3 + DC cells reduce the expression of costimulatory factors, reduce the secretion of IL-12, promote IL-10 production and inhibit the stimulation of alloreactive T cell proliferation response capacity. Foxp3 + DC cells play important roles in inhibiting corneal allograft immune response and prolonging graft survival time.

Ocular trauma score in transferred fireworks-related ocular injuries: a case series.

BACKGROUND: Our aim was to review the characteristics of transferred fireworks-related ocular damage and to evaluate the prognostic value of the ocular trauma score (OTS) for these injuries. METHODS: This study included 22 transferred patients (19 male, 3 female; mean age 22.6±14.9 years) (25 eyes). The data were retrospectively reviewed, including the characteristics of the geography, types of fireworks, status of injuries, therapeutic procedures, and the best-corrected visual acuity (BCVA). All the injured eyes were classified using the OTS at the time of the initial examination. RESULTS: Twenty eyes (80%) were in OTS category 1, three eyes (12%) were in OTS category 2, and two eyes (8%) were in OTS category 3. All cases received surgical therapy. Six eyes (24%) were enucleated, four (16%) of which achieved an improvement in their final BCVA. There was a statistically significant improvement in final BCVA between OTS category 1 and the other two OTS categories (p=0.016). CONCLUSION: The aforementioned transferred fireworks-related ocular injury cases occurred mainly in young adults, men and active participants, all of which incurred serious vision loss and blindness. The OTS is quite effective for classifying the status and estimating the prognosis of transferred fireworks-related ocular injuries.

The sphingosine-1-phosphate receptor agonist FTY720 and its phosphorylated form affect the function of CD4+CD25+ T cells in vitro.

The sphingosine-1-phosphate receptor agonist FTY720 and FTY720-P have a wide variety of fundamental functions. Many studies have demonstrated that CD4+CD25+ regulatory T (Treg) cells engage in the maintenance of immunological self-tolerance by actively suppressing self-reactive lymphocytes. Although FTY720 has also recently shown to possess an additional effect that increases the functional activity of Treg cells, the mechanism leading to the enhanced Treg activity after FTY720 treatment is still not clear. We isolated Treg cells, which were co-cultured with FTY720 or FTY720-P. The proliferation of co-cultured Treg cells was detected by the cell counting kit-8. The changes of the phenotype CD25+ and forkhead box P3 (Foxp3)+ of co-cultured Treg cells were measured by flow cytometry. The levels of IL-10 and TGF-ß1 in the supernatants were detected by Elisa. Cytokine mRNA expressions in co-cultured Treg cells were analyzed by real-time quantitative PCR. Mixed lymphocyte reaction assay examined the suppressive function. We found that neither FTY720 nor FTY720-P affected the proliferation of co-cultured Treg cells. The percentages of CD25+ and Foxp3+ were enhanced in the high-dose FTY720-P group. The levels of TGF-ß1 in the supernatants were enhanced in the high-dose FTY720 group. Medium and high-dose FTY720-P also enhanced the levels of TGF-ß1. TGF-ß1 and Foxp3 mRNA expression were upregulated in the high-dose FTY720-P group. The proliferation of effector T (Teff) cells was suppressed significantly in the medium and high-dose FTY720-P group at a Treg/Teff cell ratio of 1:1. At a ratio of 1:1, the proliferation of Teff cells was also suppressed in the high-dose FTY720 group. It can be concluded that high-dose FTY720-P can enhance the immune function of co-cultured Treg cells, and that medium-dose FTY720-P and high-dose FTY720 could partly enhance the function. The reason may be attributed to enhanced levels of TGF-ß1 and Foxp3.

Topical application of FTY720 and cyclosporin A prolong corneal graft survival in mice.

PURPOSE: To investigate the effects of topical FTY720 and cyclosporin A (CsA) on allogeneic corneal transplantation in mice. METHODS: A total of 75 BALB/c mice received corneal grafts from C57BL/6 donors. Recipients were treated with 0.1%, 0.3%, or 0.5% FTY720 ophthalmic gel or 1% CsA eye-drops after the graft (controls received no treatment). The number of cluster of differentiation (CD)4+ T cells and CD4+CD25+forkhead box P3 (Foxp3)+ regulatory (Treg) cell phenotypes were measured by flow cytometry. Cytokine mRNA expression in corneal grafts was analyzed by real-time quantitative PCR. CD4 + T cells and cytokines in corneal samples were identified by immunohistochemical staining. RESULTS: Corneal graft survival was prolonged by treatment with topical 0.5% FTY720 (mean survival time [MST], 24.1±1.6 days) or 1% CsA eye-drops (MST 25.0±1.9 days) compared with controls (MST, 13.4±0.5 days; n=9, both p<0.01). Topical 0.5% FTY720 treatment significantly increased the percentages of CD4 + T (p<0.05) and Treg cells (p<0.01; n=5) in the cervical lymph nodes compared with controls. Transforming growth factor-ß1 (TGF-ß1) mRNA transcription in corneal grafts after topical 0.5% FTY720 increased (p<0.05, n=3), while interleukin-2 (IL-2) and interferon-γ (IFN-γ) mRNA expression in corneal grafts treated with 1% CsA decreased (p<0.01, p<0.05, respectively). These cytokine results were paralleled by similar immunohistochemical staining. Topical 0.5% FTY720 and 1% CsA treatment reduced the infiltration of CD4+ Tcells in the grafts. CONCLUSIONS: Topical 0.5% FTY720 and 1% CsA can effectively prolong allogeneic corneal graft survival in mice. Treatment with topical 0.5% FTY720 increases the percentage of CD4+ T cells and the percentage of Treg cells in cervical lymph nodes. The 0.5% FTY720 increased TGF-ß1 mRNA expression and decreases infiltration of CD4+ T cells in corneal grafts, while topical 1% CsA down-regulated the expression of IL-2 and IFN-γ.

Association of Aurora-A (STK15) kinase polymorphisms with clinical outcome of esophageal cancer treated with preoperative chemoradiation.

BACKGROUND: Aurora-A/STK15 is a serine/threonine kinase critical for regulated chromosome segregation and cytokinesis. We investigated the association between 2 nonsynonymous single nucleotide polymorphisms in the coding region of STK15, T91A (Phe31Ile) and G169A (Val57Ile), and clinical outcome of esophageal cancer treated with preoperative chemoradiation. METHODS: Genotypes at Phe31Ile and Val57Ile were assessed from peripheral blood lymphocytes of 190 esophageal cancer patients and were correlated to response to treatment, recurrence rate, risk of death, disease-free survival (DFS) and median survival time (MTS). RESULTS: All patients had resectable esophageal or gastroesophageal junction cancer and received preoperative chemoradiation followed by esophagectomy. The heterozygous variant Phe31/Ile variant was significantly associated with tumor recurrence (odds ratio [OR] = 4.39; 95% confidence interval [CI], 2.12-8.94; P < .001), shorter DFS (P = .0001), and shorter MTS (P = .012). For patients receiving cisplatin-based therapy, only the variant Phe31/Ile had an adverse effect on response (OR = 2.8; 95% CI, 1.01-5.17; P = .048) and MTS (P = .026). The variant 91A-169G haplotype carried a significant risk for lack of complete response (OR = 2.54; 95% CI, 1.15-5.54) and higher rate of recurrence (OR = 2.73; 95%CI, 1.00-7.29). The presence of at least 1 variant allele at each locus further increased the risk of recurrence (adjusted OR = 6.21; 95% CI, 2.28-17.11; P = <.001), and was associated significantly shorter DFS (P = .003). CONCLUSIONS: Our study shows that functional SNPs in the STK15 gene are associated with higher rate of recurrence, higher likelihood of chemoratiotherapy-resistance, shorter DFS, and shorter MTS. Confirmation of our data and understanding the mechanisms through which STK15 functional SNPs mediate resistance to chemoradiotherapy are warranted.

Changes in objective vault and effect on vision outcomes after implantable Collamer lens implantation: 1-year follow-up.

PURPOSE: To investigate the changes in vault and the effect on visual outcomes 1 year after implantable Collamer lens (ICL) implantation. METHODS: In this retrospective study, 127 eyes of 66 patients undergoing ICL implantation were examined both before and up to 1 year after the surgery. The examination contents included white-to-white (WTW) diameter, central vault of the ICL (distance between posterior surface of ICL and anterior surface of crystalline lens), refractive error, and wavefront high-order aberration (HOA). All statistical analyses were performed using SPSS 13.0 software. RESULTS: A significant decrease in vault was noted up to 1 month, after which the value stabilized (p=0.001). The moderate vault decreased significantly after the first 3 months postsurgery (paired-samples t test, p<0.05). Low vault showed a tendency to increase and high vault showed a tendency to decrease, but not significantly, over time. There was no statistically significant correlation between the amount of vault and the refractive error (Pearson correlation coefficient R=0.111, p=0.473) and there was a statistically significant correlation between the vault and HOAs (R=0.304, p=0.024). CONCLUSIONS: Implantable Collamer lens vault over the crystalline lens had the tendency toward a slight decrease with time and did not significantly affect the vision outcome 1 year after surgery.

[Construction of lentiviral expression vector containing Foxp3 gene and its expression in DC2.4 cell line].

AIM: To construct a Foxp3 lentiviral vector and transfer it into DC2.4 cells, which provides Foxp3+DC cells for further study on its immune modulation. METHODS: We cloned mouse Foxp3 gene into lentiviral vector(pGC-FU) and acquired the plasmid pGC-FU-Foxp3. PCR and sequencing analysis were made for verifying the positive clones. The virus packaging plasmids(pGC-FU-Foxp3, pHelper1.0 and pHelper2.0) were contransfected into 293T cells, and the Lentivirus-Foxp3 was harvested from 293T cells. The Lentivirus-Foxp3 was used to infect DC2.4 cells in vitro and the expression of Foxp3 in infected DC2.4 cells was detected with flow cytometry(FCM). RESULTS: PCR and sequencing revealed that the pGC-FU-Foxp3 plasmid was correctly constructed. The Lentivirus-Foxp3 with a titer of 2×10(8); TU/mL was successfully packaged. Foxp3 expression in DC2.4 cells infected with the Lentivirus-Foxp3 was increased significantly compared with negative control lentivirus. CONCLUSION: The pGC-FU-Foxp3 plasmid has been successfully constructed and the Lentivirus-Foxp3 has been successfully packaged. Foxp3 can be enhanced in DC cells infected with the Lentivirus-Foxp3.

Experimental study of the mechanism of tolerance induction in dexamethasone-treated dendritic cells.

BACKGROUND: The aim of this study was to investigate the mechanisms underlying tolerance induction of dexamethasone (Dex)-treated dendritic cells (DCs). MATERIAL/METHODS: Well-grown DC2.4 cells were randomly assigned to receive control, 50 µg/L, 100 µg/L, or 200 µg/L of dexamethasone and then were cultured for 6 days. The expressions of CD80, CD86, galectin-9, and PD-L1 on the surface of DC2.4 cells were analyzed with flow cytometry and the level of IL-12 secreted by DC2.4 cells was determined by ELISA. The stimulating activity of DC2.4 cells on allogeneic T cells was assessed with mixed lymphocyte reaction. Dexamethasone-treated DC2.4 cells were co-cultured with allogeneic splenic lymphocytes and the Foxp3 expression in naive T lymphocytes was determined with flow cytometry. RESULTS: Compared with the control group, the expressions of CD80, CD86, galectin-9, and PD-L1 on the surface of DC2.4 cells exposed to different doses of dexamethasone showed no significant changes; however, dexamethasone treatment significantly reduced IL-12 secretion and inhibited DC2.4's stimulation on the proliferation of allogeneic T lymphocytes. Moreover, dexamethasone-treated DC2.4 cells effectively promoted FOXP3 expression in naive T lymphocytes. CONCLUSIONS: DC2.4 is a stable cell line with high expressions of CD80, CD86, and PD-L1. Dexamethasone does not significantly change the cell phenotype of DC2.4 cells, but inhibits the secretion of IL-12 cytokine and attenuates DC2.4's stimulation of the proliferation of allogeneic T cells. Dexamethasone-treated DC2.4 cells also effectively promote FOXP3 expression in naive T lymphocytes.

[Outcome after treatment of myopia with implantable Collamer lens].

OBJECTIVE: To evaluate the efficacy, predictability, stability, and safety of the surgical correction myopia using implantable Collamer lenses (ICL) in phakic eyes. METHODS: A prospective study was performed to analyze 91 eyes of 48 patients who had the implantation of ICL for the treatment of myopia from July 2008 to February 2010. Patients were examined preoperatively and followed at 1 week, 1, 3, 6, and 12 months postoperatively. The examinations included the uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), refraction, contrast sensitivity, wavefront aberration, intraocular pressure, space between crystal lens and intraocular lens (vault), endothelial cell density (ECD), anterior chamber depth (ACD), surgical complication, etc. RESULTS: Successful implantation was achieved in all patients. The mean follow-up time was (9.54 ± 4.12) months (SD)(range 1 to 12 months). The mean preoperative SE was -12.38 diopters (D) (range -5.0D to -23.0D). Postoperatively, UCVA was maintained or improved in all eyes. UCVA achieved 1.0 in 58 eyes (64%) and BCVA gained more than 1 line in 69 eyes (75.9%). The glare and no glare contrast sensitivity were improved at 3cd, 12cd and 18cd, with the exception of 6cd. The aberration decreased in RMS, spherical and coma. Post operative ACD (1 week after operation) diminished 8.92% as compared with the preoperative ACD. The mean vaulting was (452 ± 216.38) µm (6 months) and ranged 130 - 1080 µm at different follow-up periods. There was no statistically significant difference in vaulting between postoperative data at different periods (t = 0.200, P > 0.05). The mean postoperative ECD decreased but had no statistically difference compared with the preoperative ECD. ACD decreased 31% after surgery in 2 eyes (2.1%). Transient high IOP was observed in 13 eyes (2.1%) one week after the operation. CONCLUSION: These results indicate that ICL implantation in the treatment of myopia is efficient, predictable, safe, and reliable.

[Effects of anti-CD25 monoclonal antibody on the level of cytokines in aqueous humour after rat penetrating keratoplasty].

OBJECTIVE: To investigate the toxicity of anti-CD25 monoclonal antibody (mAb) to rat cornea and its effects on the cytokines in the aqueous humour after penetrating keratoplasty (PKP), thereby evaluating the effect of anti-CD25 mAb in preventing corneal allograft rejection. METHODS: The corneal toxicity of anti-CD25 mAb at 50, 100 and 200 microg administered via subconjunctival injection was evaluated in 12 SD rats by histological examination and transmission electron microscopy. Another 93 SD rats were randomized into 5 groups, and transplantation of corneal allograft from Wistar rats was performed in 4 groups with the other group as the normal control. The 4 allograft groups were treated with saline, 100 microg anti-CD25 mAb, 100 microg anti-CD25 mAb with 50 microg dexamethasone, and 50 microg dexamethasone, respectively. The graft rejection was observed, the aqueous humour levels of IFN-gamma and IL-4 were measured with ELISA, and IFN-gamma mRNA expressions in the grafts detected with RT-PCR. RESULTS: anti-CD25 mAb at 50 or 100 microg did not show significant toxicity on the cornea, but at 200 microg, the mAb caused swelling of the corneal stromal cells and endothelial cells. After corneal allograft transplantation, a significant delay in allograft rejection was observed in the 3 groups with mAb or dexamethasone treatment as compared with that in saline group (P<0.05). IFN-gamma mRNA expression in the allograft on days 11 after PKP and in the aqueous humour on days 6 and 11 was markedly increased in saline group compared with that in the 3 treatment groups (P<0.05). The mean IL-4 level in the aqueous humour was significantly higher in the mAb group than in saline group (P<0.05), but markedly lower in anti-CD25 mAb+dexamethasone and dexamethasone groups than in anti-CD25 mAb group (P<0.05). CONCLUSIONS: Anti-CD25 mAb at 20 and 100 microg does not obviously affect the rat corneas. Anti-CD25 mAb inhibits IFN-gamma expression and promotes IL-4 the expression to reduce corneal allograft rejection, whereas anti-CD25 mAb with low-dose dexamethasone inhibits both IFN-gamma and IL-4 expressions to more effectively promote the graft survival.

Polymorphisms of STK15 (Aurora-A) gene and lung cancer risk in Caucasians.

STK15/Aurora-A is a centrosome-localized serine/threonine kinase that functions primarily in centrosome maturation and mitotic spindle assembly. In a large lung cancer case-control study of 1401 cases and 1397 controls including three ethnic groups, we examined the associations between two non-synonymous SNPs (Phe31Ile and Val57Ile) of the STK15 gene and lung cancer risk. There were statistically significant differences in the distribution of the genotypes (P<0.0001) and haplotypes (P<0.0001) by ethnicity for the Phe31Ile, but not the Val57Ile variant. Caucasians with the homozygous variant Phe31Ile genotype (Ile/Ile) were at a significantly reduced risk for lung cancer [odds ratio (OR)=0.63, 95% confidence interval (CI)=0.41-0.96]. The variant allele of Val57Ile was not associated with lung cancer risk overall. However, men with the homozygous variant genotype (Ile/Ile) had a reduced lung cancer risk as compared with men with the wild-type genotype (Val/Val) (OR=0.42, 95% CI=0.19-0.94). When we performed joint analysis of these two polymorphisms, compared with the reference group (TT+GG, 40.99% of controls), homozygous Ile31 allele/wild-type Val57 allele (AA+GG) carriers (5.45% of controls) exhibited a reduced lung cancer risk (OR=0.78, 95% CI=0.63-0.97). This is the first epidemiological study to report significant associations between STK15 polymorphisms and lung cancer risk.

[Effect of IL-1 receptor antagonist on cell apoptosis in rat corneal grafts].

AIM: To observe the apoptosis of cells in rat corneal grafts at acute rejection phase and explore the effects of IL-1 receptor antagonist (IL-1ra) on cell apoptosis. METHODS: The penetrating corneal transplantation model was established. Corneal grafting was divided into four groups, namely, normal Wistar rat control group (no grafting, group A), isograft (Wistar rat-->Wistar rat, group B), allograft (Wistar rat-->SD rat) with normal saline treatment (group C) and allograft (Wistar rat-->SD rat) with IL-1ra treatment (group D). Cell apoptosis in corneal grafts was detected by TUNEL staining at 7 d, 10 d and 14 d after transplantation, and an automatic image analyzer was used to analyze the results, which were expressed as positive unit (PU). The changes of cellular ultrastructure in corneal grafts were observed under transmission electron microscope. RESULTS: (1)The average survival time of corneal grafts in C and D groups was (10.38+/-1.85) d and (13.56+/-1.94) d, respectively, with significant difference (P<0.01). (2)As compared with normal and non-rejected corneas, cell apoptosis and necrosis commonly existed in corneal grafts which rejection had occur. (3)In normal corneas, there were merely a very small number apoptotic cells in epithelial laminal, and apoptotic cells were found hardly in stromal laminal and endothelial cell layers. However, sporadic apoptotic cells were found in all layers of corneal grafts in B, C and D groups at 10 d after transplantation, the average PU being of no notably difference (P>0.05). Apoptosis obviously increased in nearby regions of wound and central area of corneal grafts in C and D groups, especially in C group. The apoptotic cells were distributed mainly in basal layer of epithelial cells and stroma of superficial layer. CONCLUSION: Cell apoptosis plays an important role in corneal graft rejection reaction. IL-1ra treatment can prolong the survival time of corneal grafts by means of suppression of cell apoptosis in corneal grafts.