[The effect of Vaccinium uliginosum to the electroretinogram and retina of rabbits before and after light-induced damage].

OBJECTIVE: Study the effect of Vaccinium uliginosum (VU) on the electroretinogram (ERG) and histology of rabbits' retina before and after light-induced damage. METHODS: It was an experimental study in contrast. Thirty-five Chinchilla Rabbits were divided into five groups according to the randomization tables. All rabbits ate and drank freely except those in group A and D who were fed with VU homogenate. Four weeks later we observed the tissue & structure of the rabbits in group D and E under the light microscope and measured the thickness of their out nuclear layers (ONL) and apoptosis index (AI). At the same time, we recorded maximal combined reaction ERG and oscillatory potentials (oscillatory potentials, OPs) of the left rabbits according to the standards set by the International Society for Clinical Electrophysiology of Vision (ISCEV). Then group A, B and C were exposed to strong light. Also we stopped feeding group A with VU and start to feed group C with it. We recorded ERG of them all after 1 day, 1 week and 2 weeks respectively. Then we observed the tissues & structures of them. SPSS 12.0 software package was used for one-way or double ways ANOVA and LSD test. RESULTS: (1) Maximal combined reaction ERG: after four weeks feeding implicit time of group A: a wave (14.079+/-0.841) ms, b wave (35.629+/-6.865) ms; amplitude: a wave (83.936+/-10.807) microV, b wave (280.931+/-27.807) microV. Two weeks after injury implicit time of group A: a wave (15.571+/-1.087) ms, b wave (38.915+/-7.683) ms, amplitude:a wave (66.478+/-9.709) microV, b wave (245.887+/-11.797) microV. After four weeks feeding implicit time of group B: a wave (15.635+/-1.661) ms, b wave (42.985+/-3.164) ms; amplitude: a wave (69.331+/-12.355) microV, b wave (197.331+/-16.157) microV. Two weeks after injury implicit time of group B: a wave (18.783+/-1.966) ms, b wave (52.322+/-4.784) ms, amplitude:a wave (57.562+/-8.217) microV, b wave (159.569+/-17.859) microV. After four weeks feeding implicit time of group C: a wave (15.115+/-0.940) ms, b wave (43.242+/-4.662) ms, amplitude: a wave (72.812+/-4.403) microV, b wave (207.815+/-14.373) microV. Two weeks after injury implicit time of group C: a wave (15.957+/-2.154) ms, b wave (44.081+/-9.506) ms, amplitude: a wave (66.804+/-8.755) microV, b wave (186.271+/-29.349) microV. These three groups had statistical significance in maximal combined reaction ERG (implicit time of a wave: fed 4 weeks F=6.057, P<0.05; two weeks after injury F=13.296, P<0.05. Implicit time of b wave: fed 4 weeks F=9.949, P<0.05; two weeks after injury F=11.145, P<0.05. Amplitude of a wave: fed 4 weeks F=8.468, P<0.05; two weeks after injury F=4.844, P<0.05. Amplitude of b wave: fed 4 weeks F=70.194, P<0.05; two weeks after injury F=62.161, P<0.05). The total amplitudes of OPs (OPs=OPs1+OPs2+OPs3) of them had statistical significance (fed 4 weeks F=17.482, P<0.05; two weeks after injury F=11.748, P<0.05). By LSD test we found that before injury the amplitude of a wave and b wave in group B and C in maximal combined reaction ERG were significantly lower than those of group A which was fed by VU for 4 weeks (the a wave and b wave of group B compared to A: P=0.003, 0.000; the a wave and b wave of group C compared to A: P=0.001, 0.000). After light-induced injury, the implicit time of all the groups was increased and amplitude was decreased. But after the injury time of 1 day, 1 week and 2 weeks, the amplitude of b wave of group A was respectively (229.743+/-11.978) microV, (212.785+/-21.021) microV, (245.887+/-11.797) microV, which was significantly higher than group B in the same period (P=0.000). With the accumulation of VU the ERG of group C was improving. Two weeks after injury the implicit time of b wave in group C was (44.081+/-9.506) ms and the amplitude was (186.271+/-29.349) microV. Compared with group B the former was decreased and the latter was increased significantly (implicit time: P=0.008; amplitude: P=0.007). (2) Group D and E were normal in histology and layers were in order. While group B got disordered. Group A and C were injured slightly. The thickness of ONL among all groups had statistical significance (F=330.506, P<0.05). (3) There was statistical significance among all groups in AI (F=230.126, P<0.05). AI of group B was (10.960+/-1.534)% and was higher than others' (P=0.000). AI of group D was (1.817+/-0.203)% and lower than group E (P=0.000). CONCLUSIONS: Vaccinium uliginosum can decrease retinal cell apoptosis and reduce photochemical damage to retinal tissue. In addition VU is able to promote retinal repairing after light damage.

[Relationship between retinal neurons apoptosis and changes of manganese superoxide dismutase activity and mRNA expression in early diabetic rats].

OBJECTIVE: To study the relationship between the apoptosis of retinal neurons and changes of manganese superoxide dismutase (MnSOD) activity and mRNA expression in early diabetic rats. METHODS: Controlled experimental study. Seventy two male 8-week-aged SD rats were divided into control group and diabetic mellitus group. Each group were subdivided into 4, 8 and 12 weeks groups (each group, n = 12). The diabetic group rats were injected with a single dose of streptozotocin (60 mg/kg) to induce the diabetic model, the control group rats were raised without any intervention. Apoptosis of retinal neurons was detected by TdT-mediated dUTP nick end label (TUNEL) assay. The protein expression of caspase-3 was detected by immunohistochemistry. Xanthine oxidase method was used to measure the activity of MnSOD and copper-znic superoxide dismutase (Cu-ZnSOD). MnSOD mRNA, Cu-ZnSODmRNA and caspase-3 mRNA levels were determined by SYBR Green Realtime PCR Master Mix. ANOVA was used to test the comparisons of the apoptosis ratio of retinal ganglion cells, the levels of caspase-3 mRNA, and the activity and mRNA levels of Cu-ZnSOD and MnSOD in the control groups and the diabetic groups, and LSD was used to test multiple comparisons. RESULTS: (1) The difference of the retinal ganglion cells apoptosis ratio in the control groups and the diabetic groups had statistical significance at 4, 8, 12 weeks (F = 19.5412, P < 0.05). There were no apoptosis neurons in control groups' retina at 4, 8, 12 weeks. Apoptosis of the retinal neurons occurred 4 weeks after the onset of diabetes. The apoptosis ratio of retinal ganglion cells in rats that had diabetes for 8 and 12 weeks was (5.7 +/- 3.9)% and (11.8 +/- 5.1)%, respectively, and was significantly higher than that of age-matched control groups (8 and 12 weeks: P = 0.000). (2) Caspase-3 protein expression was not observed in the control rats' retina at 4, 8, 12 weeks. Positive staining of caspase-3 occurred 4 weeks after the onset of diabetes, and enhanced at 8 and 12 weeks. The difference of caspase-3 mRNA levels in the control groups and the diabetic groups had statistical significance at 4, 8, 12 weeks (F = 105.175, P < 0.05). In control rats at 4, 8 and 12 weeks, caspase-3 mRNA levels were 1.649 +/- 0.586, 1.526 +/- 0.486, 1.614 +/- 0.296, respectively. Caspase-3 mRNA levels in diabetic rats that had diabetes for 4, 8 and 12 weeks were 5.672 +/- 1.193, 12.566 +/- 2.272, 14.297 +/- 2.11, respectively, which were greater than that in the control groups (4, 8 and 12 weeks: P = 0.000). (3) The difference of the activity and mRNA levels of MnSOD and Cu-ZnSOD in the control groups and the diabetic groups had statistical significance at 4, 8, 12 weeks (MnSOD: activity: F = 19.709, P < 0.05, mRNA: F = 93.352, P < 0.05; Cu-ZnSOD: activity: F = 16.708, P < 0.05, mRNA: F = 16.332, P < 0.05). In the control groups at 4, 8 and 12 weeks, the activity of MnSOD was (47.118 +/- 5.018), (46.033 +/- 6.835) and (45.813 +/- 6.859) U/mg, respectively; and MnSOD mRNA levels were 0.973 +/- 0.123, 0.974 +/- 0.085 and 0.994 +/- 0.074, respectively. The activity of Cu-ZnSOD was (113.884 +/- 9.07), (112.301 +/- 5.24) and (117.52 +/- 7.982) U/mg, respectively; and Cu-ZnSOD mRNA levels of were 1.067 +/- 0.109, 1.055 +/- 0.119, 1.092 +/- 0.180, respectively. In the rats that had diabetes for 4, 8 and 12 weeks, the activity of MnSOD was (33.863 +/- 6.909), (22.877 +/- 7.875) and (20.034 +/- 6.796) U/mg, respectively; and MnSOD mRNA levels were 0.627 +/- 0.083, 0.333 +/- 0.080, 0.256 +/- 0.057, respectively; these data were less than those in the age-matched control groups (activity: 4 weeks: P = 0.002, 8 and 12 weeks: P = 0.000; mRNA: 4, 8 and 12 weeks: P = 0.000). The activity (109.793 +/- 7.468) U/mg and mRNA level (0.976 +/- 0.108) of Cu-ZnSOD in rats had diabetes for 4 weeks showed no significant difference as compared to age-matched control group (activity: P = 0.426; mRNA: P = 0.172). In diabetic rats at 8 and 12 weeks, the activity of Cu-ZnSOD was (98.588 +/- 9.212) and (78.168 +/- 12.180) U/mg, respectively; Cu-ZnSOD mRNA levels were 0.829 +/- 0.048 and 0.621 +/- 0.033, respectively; these data were less than those in the age-matched control group (activity: 8 weeks: P = 0.011, 12 weeks: P = 0.000; mRNA: 8 weeks: P = 0.001, 12 weeks: P = 0.000). The changes of MnSOD occurred as early as 4 weeks after the onset of diabetes, while the changes of the Cu-ZnSOD occurred later, mainly at 8 and 12 weeks after the onset of diabetes. CONCLUSIONS: Apoptosis of the retinal neurons in early diabetic rats may correlate with the decline of the activity and mRNA expression of MnSOD.

[Ocular manifestations of brainstem tumor].

OBJECTIVE: To investigate the ocular manifestations of brainstem tumors and to avoid misdiagnosis and missed diagnosis. METHODS: This is a retrospective case series study. The clinical data of 57 brainstem tumor in-patients were collected from 1993 to 2007. The clinical manifestations and the results of related examinations were analyzed. RESULTS: The present series included 51 cases of brainstem germinoma, 4 cases of cavernous hemangioma, 1 case of hemangioblastoma and 1 case of metastatic tumor. In 51 cases of brainstem germinoma, there were 37 males and 14 females. The first attack age varied from 5 to 55 years old and the median age was 23 years old. The high incident ages of brainstem germinoma were 10 - 35 years. Patients were presented with diplopia, ocular motility disturbance, nystagmus, anisocoria, and facial palsy. In 57 patients, diplopia was the initial symptom in 12.3% (7/57) cases. The incidence of oculomotor nerve paralysis was 17.5% (10/57); trochlear paralysis, 1.8% (1/57); trigeminal nerve paralysis, 5.3% (3/57); abducens nerve paralysis, 35.1% (20/57); facial palsy, 14.0% (8/57); optic disc edema, 19.3% (11/57); nystagmus, 21.1% (12/57) and anisocoria, 10.5% (6/57). CONCLUSIONS: Ocular manifestations occur frequently in brainstem tumor. Nuclear ophthalmoplegia, nystagmus and other neuro-ophthalmic signs provide helpful clues for the diagnosis of brainstem tumor.

Valproic acid treatment inhibits hypoxia-inducible factor 1α accumulation and protects against burn-induced gut barrier dysfunction in a rodent model.

OBJECTIVE: Burn-induced gut dysfunction plays an important role in the development of sepsis and multiple organ dysfunction. Emerging evidence suggests that hypoxia-inducible factor-1α (HIF-1α) is critical in paracellular barrier functions via regulating vascular endothelial growth factor (VEGF) and myosin light chain kinase (MLCK) expression. Previous studies have also demonstrated that histone deacetylase inhibitors (HDACIs) can repress HIF-1α. This study aims to examine whether valproic acid (VPA), a HDACI, protects against burn-induced gut barrier dysfunction via repressing HIF-1α-dependent upregulation of VEGF and MLCK expression. METHODS: Rats were subjected to third degree 55% TBSA burns and treated with/ without VPA (300 mg/kg). Intestinal barrier dysfunction was evaluated by permeability of intestinal mucosa to fluorescein isothiocyanate (FITC)-dextran and histologic evaluation. Histone acetylation, tight junction protein zonula occludens 1 (ZO-1), VEGF, MLCK and HIF-1α were measured. In addition, CaCO2 cells were transfected with siRNA directed against HIF-1α and were stimulated with CoCl2 (1mM) for 24 hours with/without VPA (2mM) followed by analysis of HIF-1α, MLCK, VEGF and ZO-1. RESULTS: Burn insults resulted in a significant increase in intestinal permeability and mucosal damage, accompanied by a significant reduction in histone acetylation, ZO-1, upregulation of VEGF, MLCK expression, and an increase in HIF-1α accumulation. VPA significantly attenuated the increase in intestinal permeability, mucosa damage, histone deacetylation and changes in ZO-1 expression. VPA also attenuated the increased VEGF, MLCK and HIF-1α protein levels. VPA reduced HIF-1α, MLCK and VEGF production and prevented ZO-1 loss in CoCl2-stimulated Caco-2 cells. Moreover, transfection of siRNA directed against HIF-1α led to inhibition of MLCK and VEGF production, accompanied by upregulation of ZO-1. CONCLUSIONS: These results indicate that VPA can protect against burn-induced gut barrier dysfunction. These protective effects may be due to its inhibitory action on HIF-1α, leading to a reduction in intestinal VEGF and MLCK expression and minimizing ZO-1 degradation.

Ulinastatin suppresses burn-induced lipid peroxidation and reduces fluid requirements in a Swine model.

Objective. Lipid peroxidation plays a critical role in burn-induced plasma leakage, and ulinastatin has been reported to reduce lipid peroxidation in various models. This study aims to examine whether ulinastatin reduces fluid requirements through inhibition of lipid peroxidation in a swine burn model. Methods. Forty miniature swine were subjected to 40% TBSA burns and were randomly allocated to the following four groups: immediate lactated Ringer's resuscitation (ILR), immediate LR containing ulinastatin (ILR/ULI), delayed LR resuscitation (DLR), and delayed LR containing ulinastatin (DLR/ULI). Hemodynamic variables, net fluid accumulation, and plasma thiobarbituric acid reactive substances (TBARS) concentrations were measured. Heart, liver, lung, skeletal muscle, and ileum were harvested at 48 hours after burn for evaluation of TBARS concentrations, activities of antioxidant enzymes, and tissue water content. Results. Ulinastatin significantly reduced pulmonary vascular permeability index (PVPI) and extravascular lung water index (ELWI), net fluid accumulation, and water content of heart, lung, and ileum in both immediate or delayed resuscitation groups. Furthermore, ulinastatin infusion significantly reduced plasma and tissue concentrations of TBARS in both immediate or delayed resuscitation groups. Conclusions. These results indicate that ulinastatin can reduce fluid requirements through inhibition of lipid peroxidation.

The effect of Vaccinium uliginosum on rabbit retinal structure and light-induced function damage.

OBJECTIVE: To study the effect of Vaccinium uliginosum L., (VU) on the electroretinogram (ERG) and retinal pathological changes in rabbits after light-induced damage. METHODS: Twenty-eight Chinchilla rabbits were randomly divided into four groups: administration beforehand (A), administration after injury (B), light injury without administration (C), and blank (D) groups. After a 4-week administration of VU homogenate at 4.8 g/(kg·d) once a day in group A, ERG in groups A, B and C were recorded according to the standards set by the International Society for Clinical Electrophysiology of Vision (ISCEV). Except for group D, the groups were then exposed to strong light. Just after that, group A stopped receiving VU treatment and group B started to receive it. Then ERGs in all groups were recorded after 1 day, 1 week, and 2 weeks. Throughout the whole process groups which were not fed with VU were fed with normal saline. Finally, the tissues and structures of all the groups were observed and the thickness of the outer nuclear layers (ONL) was measured. RESULTS: (1) After 4-week feeding with VU, the latency time of ERG in group A became shorter than those in the other groups and the amplitude increased. After being exposed to strong light, the latency time lengthened and amplitude decreased in all the injury groups, but comparing at each time point, the measured values in group A were better than those in group C. With the accumulation of VU, the ERG in group B improved, and finally, all of the detected values became better than those in group C. (2) Retinae in group D were normal in histology and the layers were in order but those in group C became disarranged. The injuries in groups A and B were minor compared with those in group C. The thickness of the ONL in group C was significantly thinner than in the other groups (P=0.000), and that in groups A and B was thicker than that in group C, although thinner than in group D. That in group A was thicker than in group B. CONCLUSIONS: VU can relieve the injury to rabbit retinae exposed to normal day and night rhythm, alleviate the harm caused by light when used beforehand, and repair the light damage to the retina.

Effect of amnion membrane transplantation on corneal neovascularization in 10 patients with alkali burn.

By observing clinical cases, we studied the curative effect of amnion membrane transplantation on decreasing corneal neovascularization (CNV). It was a non-randomized retrospective case-control study. Among 17 cases (21 eyes) of third-degree alkali burns from 2007 to 2010, 10 cases (12 eyes) were performed with amnion membrane transplantation operation, and others were not. Amnion membrane transplantation was performed at the 3(rd) day after burn in the treatment group. Areas of CNV in double groups were measured at the 14(th) day and 60(th) day after burn. Area of CNV in the treatment group was (66.207±7.251)mm(2) at the 14(th) day after burn, and was 18.27% lower than that in the control group. Area of CNV in the treatment group was (120.046±13.812)mm(2) at the 60(th) day after burn, and was 11.35% lower than that in the control group. There was both statistical significance (P<0.05). Amnion membrane transplantation operation can inhibit the growth of corneal neovascularization induced by alkali burn.