Identification of novel prognosis-related genes in the endometrial cancer immune microenvironment.

The incidence of endometrial cancer is increasing each year, and treatment effects are poor for patients with advanced and specific subtypes. Exploring immune infiltration-related factors in endometrial cancer can aid in the prognosis of patients and provide new immunotherapy targets. We downloaded immune metagene and functional data of patients with different subtypes of endometrial cancer from The Cancer Genome Atlas database and selected the lymphocyte-specific kinase (LCK) metagene as a representative genetic marker of the immune microenvironment in endometrial cancer. The results showed that LCK metagene expression is related to the prognosis of patients with endometrioid endometrial adenocarcinoma subtypes and highly correlated with the PTEN and PIK3CA mutational status. A search for LCK-related modules returned seven independent genetic predictors of survival in patients with endometrial cancer. The TIMER algorithm showed that the expression of these seven genes was positively correlated with the infiltration levels of six types of immune cells. The diagnostic value of these markers was validated using real-time quantitative PCR and immunohistochemical methods. Our results identified CD74, HLA-DRB5, CD52, HLA-DPB1 and HLA-DRB1 as possible valuable genetic markers for the diagnosis and prognosis of endometrial cancer and provided a theoretical basis for immunotherapy targets for its clinical treatment.

[Effects of simvastatin on expression of connective tissue growth factor in vitreous and retina of diabetic rats].

OBJECTIVE: To study the effects of simvastatin on the expression of connective tissue growth factor (CTGF) that cause fibrosis in the vitreous and retina after intravitreal injection in diabetic rats, and to explore the safety of this procedure. METHODS: It was an experimental study. Forty adult male Wistar rats were randomly divided into normal control group (10 rats) and diabetes mellitus group (30 rats). Four months later, according to whether treated with simvastatin or not, the diabetes mellitus group randomly divided into simvastatin intervention group (20 rats) and diabetic positive control group (10 rats). Simvastatin was injected into the vitreous in the simvastatin intervention group, but not in the diabetic positive control group. Seven days later, after the examination of electroretinogram (ERG), all rats were sacrificed, and their eyeballs were enucleated. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry method were performed to determine the expression of CTGF in the vitreous and retina. Terminal DNA transferase-mediated dUTP nick end labeling (TUNEL) method was used to detect apoptosis of retina cells. Concentration of CTGF in the vitreous, retinal expression of CTGF, retinal cellular apoptosis index, ERG-b wave and oscillatory potentials (OPs) of rats in each group were compared using analysis of variance LSD test methods. RESULTS: No systemic toxic reactions, no lens opacity and no endophthalmitis occurred after injection of simvastatin into the vitreous of rats. Concentrations of CTGF in vitreous of simvastatin intervention group, diabetes positive control group and normal control group rats were 359.21 µg/L, 478.47 µg/L and 210.78 µg/L, respectively (F = 252.366, P < 0.05). The levels of CTGF in the vitreous of simvastatin intervention group and diabetic positive control group was significantly higher than that of the normal control group and showed significant difference (t = 12.123, 23.816;P < 0.05). CTGF levels in simvastatin intervention group were significantly lower than those in diabetic positive control group, and the difference was statistically significant. (t = 11.693, P < 0.05). The results of immunohistochemical staining and TUNEL staining were negative in the normal control group. Retinal expression of CTGF in simvastatin intervention group and diabetic positive control group were (26.60 ± 2.95)% and (42.31 ± 2.59)%, respectively. Retinal expression of CTGF in simvastatin intervention group was reduced as compared to the diabetic positive control group, the difference was statistically significant (t = 12.112, P < 0.05). Retinal cellular apoptosis index of simvastatin intervention group and diabetic positive control group was 0.19 ± 0.02 and 0.25 ± 0.03, respectively. Retinal cellular apoptosis index of simvastatin intervention group was significantly lower than that in diabetic positive control group (t = 4.745, P < 0.05). ERG revealed no significant changes. In simvastatin intervention group as compared with diabetic positive control group. CONCLUSIONS: Simvastatin could inhibit the expression of CTGF in the vitreous body and retina in diabetic rats. Intravitreal injection of simvastatin is a relatively safe procedure.

[Retinal cell apoptosis, microvascular changes and expression of connective tissue growth factor in experimental diabetic rats].

OBJECTIVE: To study the cell apoptosis and the expression of connective tissue growth factor (CTGF) in the retina of diabetic rats and to explore their contributions to the changes of microcirculation. METHODS: It was a experiment study. Fifty-five adult male Wistar rats were divided into two groups, normal control group (CON, 10 rats) and diabetes mellitus group (DM, 45 rats). The 30 surviving rats in the DM group were further divided into 3 groups based on the time of observation, 2 month (DM2), 4 month (DM4) and 6 month (DM6) groups, with 10 rats in each group. Cell apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL). Expression of CTGF was determined by immunohistochemical study. Retinal vessels were observed by retinal digest stretched periodic acid Schiff (PAS) staining. RESULTS: Immunohistochemical staining and TUNEL staining revealed negative results in normal control group. Retinal cell apoptosis index increased gradually from DM2 to DM6, the differences between any two groups were statistically significant (t(2-4, 2-6, 4-6) = 21.432, 50.843, 29.410; P < 0.05). Expression of CTGF in the retina increased from DM2-DM6, the differences between any two groups were statistically significant (t(2-4, 2-6, 4-6) = 15.345, 26.316, 10.971; P < 0.05). PAS staining of retinal blood vessels obtained negative results in the CON and DM2 groups. Part of retinal capillaries were slightly stiff and narrow in DM4 group. Retinal capillaries in DM6 group were trunk stiff and were narrowed obviously. The number of pericytes was reduced in DM4, and progressed following the course of diabetes. The number of pericytes in the DM2 group did not different from that in the CON group (t = 0.875, P = 0.387). The number of pericytes in the DM4 and DM6 group were significantly decreased as compared to the CON group (t = 3.367, 6.667; P < 0.05). Retinal cellular apoptosis index had a significant positive correlation to the expression of CTGF (r = 0.958, P < 0.05). Number of pericytes was significantly correlated (negative correlation) with retinal cellular apoptosis index and the expression of CTGF (r = -0.540, -0.595; P < 0.05). CONCLUSIONS: The appearances of cellular apoptosis and fibrosing factor CTGF in the retina of diabetic rats occurred earlier than the changes of microcirculation and the number of capillary pericytes.

[Experimental study on treatment of rabbits optic nerve injury with Caspase-3 inhibitor z-DEVD-fmk].

OBJECTIVE: To observe the effects of caspase-3 inhibitor z-DEVD-fmk on optic nerve injury of rabbits. METHODS: It was an experimental study. Two to three month-old rabbits were used in this study. The rabbit model of optic nerve injury was created by fluid percussion brain injury device (FPI). DMSO (5 µl 2% solution) was injected intravitreally to the left eyes (control group). Caspase-3 inhibitor z-DEVD-fmk (5 µl) was injected intravitreally to the right eyes (experimental group). Flash-visual evoked potential (F-VEP) and histopathological examination of the retina were used to check the variations in optic nerve injury at 1, 4, 7, 10, 14, and 21 days after the treatment. Immunohistochemistry was used to detect the expression of caspase-3 in the retina. T-test, variance analysis, q-test and linear correlation analysis were used to analyze these data. RESULTS: At the 7th day after treatment, the latency of F-VEP P1 in experimental group was shorter than that in the control group [(90.50±7.61) ms vs (113.59±12.92) ms, t=4.060, P<0.05] and the number of retinal ganglion cells (RGC) in the experimental group was greater than that in the control group (237.62±8.50 vs 207.03±11.04, t=-5.843, P<0.05). Both of these trends continued to the 21st day after treatment [(67.97±7.93) ms vs. (134.22±8.50) ms, t=13.950, P<0.05; 156.32±8.45 vs. 207.13±12.21, t=-10.307, P<0.05]. The absorbency (A) of caspase-3 in the experimental group (0.396±0.023) was lower than that in the control group (0.458±0.024) and this difference was statistically significant (t=6.200, P<0.05) at the 7th day after treatment. The latency of F-VEP P1 and the absorbency of caspase-3 in the retina were positively correlated with each other (r=0.95, P<0.05). CONCLUSION: z-DEVD-fmk is effective in treating rabbit optic nerve injury by inhibiting the expression of caspase-3 in the retina. It can promote the recovery of optic nerve function.

[Treatment of postoperative endophthalmitis following cataract surgery without intraocular lens removal].

OBJECTIVE: To explore the effects of vitrectomy combined with silicone oil injection on the treatment of postoperative endophthalmitis following cataract surgery without intraocular lens removal, and to analyze the relative factors. METHODS: This was a retrospective case series study. Clinical data of 7 eyes of 7 patients with postoperative endophthalmitis following cataract surgery underwent the treatment of vitrectomy combined with silicone oil injection without intraocular lens removal from 2003 to 2008 were collected. The outcomes of vision, slit lamp examination, direct and indirect ophthalmoscopy, IOP, and B-scan were observed, and the surgical effects were analyzed. RESULTS: Five patients were male, and 2 patients were female. The age ranged from 67.0 to 84.0 years with a mean of 70.0 + or - 4.5. The onset of endophthalmitis ranged from 1 to 3 days with a mean of 2 days. Silicone oil was removed in 5 eyes 3 to 6 months postoperatively. Preoperative visual acuity ranged from non light perception to hand moving. The mean preoperative intraocular pressure (IOP) was (35.0 + or - 0.5) mmHg with a range from 35.0 to 56.0 mmHg. Follow-up periods ranged from 6 to 43 months with a mean of (10 + or - 6) months. Postoperative visual acuity ranged from non light perception to 0.8. Postoperative vision increased in 6 eyes (86%), and was stable in 1 eye (14%). The mean postoperative IOP was (18.0 + or - 1.5) mmHg with a range from 10.0 to 20.0 mmHg, this was significantly lower than that preoperatively (t = 1.94, P < 0.05). Postoperative complications mainly included fibrous exudates in the anterior chamber at early stage after the surgery (7 eyes) and temporary intraocular pressure elevation (1 eye). There was no retinal detachment and atrophia bulbi. CONCLUSION: Vitrectomy combined with silicone oil injection may be a safe and effective method in treating postoperative endophthalmitis following cataract surgery without intraocular lens removal.

[The expression of vascular endothelial growth factor of vitreous in patients with proliferative diabetic retinopathy].

OBJECTIVE: To explore the expression of vascular endothelial growth factor of vitreous in patients with proliferative diabetic retinopathy, and to analyze the relative factors. METHODS: It was a case-control study. Data of 50 eyes of 50 PDR patients who underwent vitrectomy were retrospectively analyzed. The vitreous fluid samples were obtained during surgery. The VEGF level in vitreous fluid was determined by enzyme linked immunosorbent assay, and the expression of vascular endothelial growth factor of vitreous in patients with proliferative diabetic retinopathy was analyzed. The mean follow-up was 9 months with a range from 6 to 26 months. The VEGF levels in vitreous fluid were compared between PDR patients and normal control, and silicone oil tamponade group and without silicone oil tamponade group by t-test. The changes of VEGF levels in vitreous fluid in eyes with regression, stabilization, and progression were analyzed by analysis of variance. The effects of VEGF levels in vitreous fluid on PDR were analyzed by analysis of variance. RESULTS: The mean VEGF level of vitreous was (592.4801 +/- 587.4267) ng/L in PDR patients, and it was (131.3022 +/- 26.9192) ng/L in normal control. VEGF level was significantly higher in PDR patients than that in normal control (t = 3.2315, P < 0.05). There were 10 eyes (20%) with progression of PDR, 10 eyes (20%) with stabilization, and 30 eyes (60%) with regression. Vitreous level of VEGF was significantly higher in eyes with progression of PDR than that in eyes with stabilization or regression of PDR (q = -3.3187, -4.0843; P < 0.05). Vitreous level of VEGF was significantly higher in eyes without retinal photocoagulation than that in eyes underwent panretinal photocoagulation or local photocoagulation (q = -4.2187, -3.9672; P < 0.05). CONCLUSION: There is a correlation between the expression of VEGF of vitreous and the severity of PDR. The expression of VEGF of vitreous presents a relatively lower level in patients with stabilization and regression of PDR.