ObjectiveTo investigate the effects and mechanisms of G protein-coupled receptor 91 (GPR91) on blood-retinal barrier (BRB) in diabetic rats. MethodsA lentiviral vector of shRNA targeting rat GPR91 and scrambled shRNA were constructed. Healthy male Sprague-Dawley (SD) rats were selected in this study. The 60 rats were randomized into 4 groups and treated as follows:(1) control group (Group A, n=15), the rats received injections of an equal volume of 0.1% citrate buffer; (2) streptozocin (STZ) group (Group B, n=15), the rats received injections of STZ; (3) LV.shScrambled group (Group C, n=15), diabetic rats received an intravitreal injection of 1 μl 1×108 TU/ml scrambled shRNA lentiviral particles at 2 weeks after the induction of diabetes; (4) LV.shGPR91 group (Group D, n=15), diabetic rats received an intravitreal injection of 1 μl 1×108 TU/ml pGCSIL-GFP-shGPR91 lentiviral particles. At 12 weeks after intravitreal injection, immunohistochemistry and Western blot were used to assess the expression of GPR91, p-extracellular signal-regulated kinase(ERK)1/2, t-ERK1/2, p-Jun N-terminal kinase (JNK), t-JNK, p-p38 mitogen-activated protein kinase (MAPK) and t-p38 MAPK. Haematoxylin and eosin (HE) staining and Evans blue dye were used to assess the structure and function of the retinal vessel. Immunohistochemistry enzyme-linked immunosorbent assay (ELISA) was used to test the protein level of VEGF. ResultsImmunohistochemistry staining showed that GPR91 was predominantly localized to the cell bodies of the ganglion cell layer. Western blot showed that GPR91 expression in Group D decreased significantly compared with Group C (F=39.31, P < 0.01). HE staining showed that the retina tissue in Group B and C developed telangiectatic vessels in the inner layer of retina, while the telangiectatic vessels attenuated in Group D. It was also demonstrated in Evans blue dye that the microvascular leakage in Group D decreased by (33.8±4.11)% compared with Group C and there was significant difference (F=30.35, P < 0.05). The results of ELISA showed the VEGF secretion of Group B and C increased compared with Group A and the VEGF expression in Group D was significantly down regulated after silencing GPR91 gene (F=253.15, P < 0.05).The results of Western blot indicated that compared with Group A, the expressions of p-ERK1/2, p-JNK and p-p38 MAPK were significantly upregulated (q=6.38, 2.94, 3.45;P < 0.05). Meanwhile, the activation of ERK1/2 was inhibited by GPR91 shRNA and the difference was statistically significant (F=22.50, P < 0.05). ConclusionsThe intravitreal injection of GPR91 shRNA attenuated the leakage of BRB in diabetic rats. GPR91 regulated the VEGF release and the leakage of BRB possibly through the ERK1/2 signaling pathway.
目的 观察运用两种不同缝线固定修补材料对疝修补术后的复发、切口感染、慢性疼痛等并发症发生情况。方法 对2008年4月至2010年4月期间笔者所在科室收治的250例腹股沟疝患者行无张力疝修补手术时,采用多股丝线或可吸收合成缝线固定修补材料进行前瞻性对比研究。结果 2组患者术后疝复发、切口感染和切口疼痛(包括慢性疼痛)发生率间的差异均无统计学意义(P>0.05)。结论 腹股沟疝无张力修补术后的复发、切口感染、慢性疼痛等并发症的发生与缝线选择无关。术者的操作技巧、严格的无菌操作原则、彻底止血以及组织损伤小才是防止术后感染、慢性疼痛等并发症发生的重要因素。
Diabetic retinopathy (DR) is the most common cause of preventable blindness in the working-age population. In addition to optimizing the hyperglycemia, hypertension, hyperlipidemia and other risk factors, regular fundus examination is essential for early diagnosis asymptomatic DR and timely treat the sight-threatening DR, so as to reduce blindness and severe visual impairment caused by DR. Clinical practice guidelines for the screening and management of DR have been implemented throughout the world, but there are reasonable differences between existing guidelines in the recommended timing of first retinal examination, screening intervals, methods for examination and criteria for referral to an ophthalmologist. It is of great clinical significance to have a detailed understanding of the current guidelines for DR screening and their clinical basis.
The neuroretinal injuries of diabetic retinopathy (DR) include retinal neuronal damage and reactive gliosis, both of which are induced by hyperglycemia and presented as early features of DR. They promote to develop mutually and accelerate the progression of DR. The molecular mechanisms study of neuronal damage mainly focuses on the alterations of extracellular environment and related signaling pathways, include inflammation, oxidative stress, endoplasmic reticulum stress, the formation of advanced glycation end products, glutamate toxicity and so on. These alterations mainly result in neuronal apoptosis and autophagy. The damaged neurons activate the glial cells with apparent changes in morphology, cell counts and the level of intracellular protein expression. In non-proliferative DR, glial cells are moderately hypertrophic and slightly increased in numbers. In proliferative DR, there is a significant rise in glial cell number with enhanced level of inflammatory factors and vascular active substances which lead a further neuronal damage. Signaling pathways of extracellular signal-regulated kinase 1/2, c-Fos and p38 mitogen-activated protein kinase are associated with their activation. Researches on the molecular mechanisms and signaling pathways of the DR will promote controlling the DR progression at the cellular level.
ObjectiveTo compare the imaging characteristics and detection of various types of lesions in diabetic retinopathy (DR) with colorful laser scanning fundus imaging (MSLI) and traditional color fundus photography (CFP).MethodsProspective case series observational study. A total of 38 eyes of 38 patients with DR diagnosed by clinical examination were included in the study. Among them, 21 were male and 17 were female; the mean age was 62.6±11.2 years; the average duration of diabetes was 14.3±7.5 years. All the patients were performed CFP, MSLI, frequency domain optical coherence tomography (SD-OCT), fluorescein angiography (FFA) examination. Using the Helielberg Spectralis HRA+OCT MSLI inspection, one scan simultaneously obtained 488 nm blue reflection (BR), 515 nm green light reflection (GR), 820 nm infrared light reflection (IR), and multicolor image (MC). The detection of traditional CFP and MC on microaneurysm (MA), hard exudation (HEX), cotton plaque (CWS), intraretinal hemorrhage (IRH), intraretinal microvascular abnormality (IRMA), venous bead (VB), venous ring (VL), macular edema (DME), macular anterior membrane (MEM) and laser photocoagulation (LB) were comparatively observed. The results of FFA examination were used as the diagnostic criteria for lesions. SD-OCT was used to determine the location and depth of lesions and the diagnostic reference for DME and MEM.ResultsThe numbers of eyes with MA (χ2=10.460), DME (χ2=4.006), MEM (χ2=4.444) was significantly higher in MC than that of traditional CFP. But the number of eyes with IRH (χ2=0.103), CWS (χ2=1.515), HEX (χ2=0.227), IRMA (χ2=0.051), VB (χ2=0.001), VL (χ2=0.149), VH (χ2=0.693) and LB (χ2=0.720) were not statistically significant between two methods (P>0.05). The imaging quality of MSLI mode is obviously better than that of traditional CFP. Among them, GR imaging shows the best structural changes of superficial retina in MA, CWS, HEX, MEM, etc. IR imaging shows clear depth in deep retina such as LB. DME was green on MC and the weak low-reflection dark area was visible on the IR image, which were consistent with the DME range indicated by the SD-OCT examination.ConclusionsCompared with the traditional CFP, the MSLI can clearly show the DR lesion. The number of checkouts is high on MA, DME and MEM by MC image.