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Objective To investigate the value of corneal laser confocal microscope in the diagnosis of corneal subepithelial plexus,corneal cell density and morphological changes of patients with diabetic retinopathy (DR).Methods Together 94 cases of confirmed DR (114 eyes),including 41 cases of non-proliferative diabetic retinopathy (NPDR group,52 eyes) and 53 cases of proliferative diabetic retinopathy (PDR group,62 eyes) were selected from January 2016 to April 2017,and meanwhile,40 diabetic patients (40 eyes) with no fundus abnormality were grouped as control group.Corneal laser confocal microscopy was used to compare the corneal subepithelial plexus,corneal cell density and morphological changes in the three groups,Results The cell densities of the basal layer,the superficial stromal layer,the medium stromal layer and the deep stromal layer of the cornea in NPDR and PDR were significantly lower than those of the control group (all P ＜ 0.05),and the PDR group was significantly lower than the NPDR group (all P ＜ 0.05).The corneal endothelial cell density,hexagonal cell ratio,nerve fiber density,and nerve fiber length in the NPDR and PDR group were significantly lower than those in the control group (all P ＜ 0.05);and the variability of endothelial cell and nerve branch density in NPDR and PDR patients were significantly higher than those in the control group (all P ＜ 0.05).The corneal endothelial cell density,hexagonal cell ratio,nerve fiber density,and nerve fiber length in the PDR group was (1962.0-± 117.3) · mm-2,46.1％ ± 5.5％,(15.4 ± 3.3) · mm-2,(6.2 ± 2.7) mm · mm-2,respectively,which were significantly lower than those in the NPDR group [(2381.4 ± 144.0) · mm-2,58.2％ ±7.0％,(20.6 ±3.8) ·mm-2,(8.6 ± 2.4)mm · mm-2,respectively] (all P ＜ 0.05),but the variability of endothelial cell and nerve branch density in PDR group were significantly higher than those in the NPDR group (all P ＜ 0.05).Conclusion Corneal confocal microscopy can effectively observe the density and morphological changes in corneal subepithelial plexus and corneal cell in DR patients so as to provide guidance for clinical diagnosis and treatment.
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Objective:To elucidate the effect of hsa-microRNA-218(hsa-mir-218)on exogenous granulysin (GLS) expression in 293T cells.Methods:Total RNA was extracted from THP-1 cells induced by phorbol 12-myristate 13-acetatefor (PMA), and GLS gene was amplified by RT-PCR, and then cloned into pDsRed-Express-C1 to construct the GLS expression vector pDsRed-GLS.Then 293T cells were co-transfected with pDsRed-GLS and pGenesil-mir-218 (pGenesil-mir-control) and laser confocal microscopy was per-formed 36 h later to detect their co-expression .Total RNA and protein were extracted 48 h post transfection , and RT-PCR and Western blot were performed to detect the effect of hsa-mir-218 on exogenous GLS expression .Results:The GLS expression vector pDsRed-GLS was constructed successfully and laser confocal microscopy indicated that it was co -expressed with the interference vector .Compared with that of cells transfected with pGenesil-mir-control, Western blot showed a markedly decrease of GLS protein expression (50%) in the cells transfected with pGenesil-mir-218.However, GLS mRNA expression remained unchanged .Conclusion: hsa-mir-218 nega-tively regulates GLS expression at a post-transcriptional level , and this provides an experimental basis for future study of mechanism of GLS expression regulated by mir-218 .
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Objetivo: describir los hallazgos de la microscopia confocal de barrido láser y su relación con la morfología de la bula de filtración.Métodos: estudio observacional descriptivo de corte transversal en 100 ojos. Se les realizó microscopia confocal de barrido láser con el HRT II y el módulo corneal de Rostock. Se aplicó la prueba de significación estadística de Mc. Nemar para una p=0,05.Resultados: predominó el grupo entre 61 y 80 años de edad (55,8 porciento) y el color de la piel blanca (46,8 porciento). En el grupo de descenso de la presión intraocular en más del 30 porciento se ubicó la mayor cantidad de bulas de todos los tamaños. Las bulas de mediano tamaño fueron las que más disminuyeron las cifras de presión intraocular, con 24 ojos (55,8 porciento, p=0,00), seguidas por las de pequeño tamaño (p=0,14). Las bulas de filtración aplanadas fueron las más frecuentes en 55 porciento de los casos (p=0,00), y 67,4 porciento de estas se ubicaron en el grupo de descenso de presión intraocular de más de 30 porciento (p=0,01). Las bulas medianas presentaron la mayor cantidad de estroma en malla porosa (60 porciento) y de microquistes epiteliales (56 porciento) p=0,00.Conclusiones: la configuración aplanada y el tamaño mediano de la bula de filtración se relacionaron con la presencia de variables histológicas que infieren buen funcionamiento de la bula (microquistes epiteliales y estroma en malla porosa). También se relacionaron con un mayor descenso de la presión intraocular
Objective: to describe the findings in the confocal laser scanning microscopy and its relation with the morphology of the filtering bleb. Methods: cross-sectional, observational and descriptive study of 100 eyes that underwent confocal laser scanning microscopy with the Retinal Heidelberg Tomography and the Rostock corneal module. Mc NemarÆs statistical significance test was made to obtain p=0,05. Results: the 61-80 years old age group (55,8 per cent) and the caucasians predominated. The group of eyes with over 30 per cent decrease of the intraocular pressure showed the highest amount of filtering blebs of all sizes. The medium-size blebs were the ones that decreases the IOP in a higher percentage (24 eyes; 55,8 per cent) for p=0,00, followed by the small blebs (p=0,14). The flat blebs were predominant in 55 per cent of cases (p=0,00) and the 67,4 per cent of them were consistent with the major range of decrease of the intraocular pressure (p=0,01). The medium size blebs presented the major number of stromas in porous mesh (60 per cent), and of epithelial mycrocists (56 per cent) for p=0,00. Conclusions: the flat configuration and the medium size of the blebs were related with the presence of the histological variables that infer adequate function of the filtrating blebs (stroma in porous mesh and epithelial mycrocists). They were also associated with higher decrease of intraocular pressure
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Objective To establish the co-culture model in vitro and induce bone marrow mesenchymal stem cells (MSCs) to differentiate into lung alveolar epithelial cells. Methods Each group had 6 samples, control group was MSCs alone; Group A was the MSCs cultured with the cells from normal lung; and Group B was the MSCs with the cells from injuried lung. Each group was cultured for 8 days and the two markers of lung alveolar epithelial cells including AQP5 and SP-C were tested by laser confocal microscopy and RT-PCR. Results Only AQP5 was detected in the control group and Group A, both AQP5 and SP-C were detected in Group B, the AQP5 mRNA expression in Group B was significantly increased compared with that in the control group(P<0.01). The AQP5 mRNA expression in Group B was also significantly increased compared with that in Group A (P<0.01). But there was no significant difference in AQP5 mRNA expression between Group A and control group. Conclusion We have successfully established the co-culture model in vitro to induce bone marrow mesenchymal stem cells to differentiate into lung epithelial cells.
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Objective To study the protective mechanism of Gross Saponin Tribulus Terrestris(GSTT) on the neonatal rat ventricular cardiocytes injured by hypoxia and the effect on protein kinase C(PKC).Methods The hypoxia-ischemia model was performed by treating the cultured neonatal rat ventricular cardiocytes with NaCN.The effect of GSTT(100 and 30 mg?L~(-1)) on the contents of ?PKC and ?PKC were detected by flow cytometry and laser confocal microscopy system.Results GSTT up-regulated ?PKC and ?PKC expressions.The contents of ?PKC and ?PKC in GSTT 100 mg?L~(-1) group(1 325.00?53.25,810.55?36.89;66.22?6.23,40.12?2.21) were increased significantly than those in model group(792.00?32.36,492.40?30.15;32.70?2.78,29.28?4.82)(P
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Objective To investigate the protective effect of glycine (Gly) on hypoxic injury to murine cardiomyocytes and the mechanisms. Methods The survival rate of cardiomyocyte survival was detected by trypan blue exclusion and lactate dehydrogenase (LDH) with an ultraviolet spectrophotometer. Ca 2+ changes in the cardiomyocytes were detected by laser confocal microscopy. Results Glycine markedly improved the survival rate of cardiomyocytes and decreased the release of LDH in cardiomyocytes after hypoxia. The protective effect was in a dose-dependent manner. Glycine could also block calcium overload after hypoxia. Conclusion Glycine has the protective effect on cardiomyocytes through the improvement of survival rate, decrease of LDH release, and blockage of calcium overload after hypoxia.
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Objective To study the quantitative changes of gap junction (GJ) in the detrusors of unstable bladder(USB) in rats,and to deduce the functional changes of GJ,which mediates intercellular communication,so as to demonstrate one of the mechanisms of USB. Methods Thirteen Wistar rats, which were identified to have USB by manometry of filling bladder after establishment of bladder outlet obstruction (BOO) model, served as study,ie,USB group.Another 10 healthy female Wistar rats served as control group.The content and distribution of connexin (Cx) 43 of the detrusors, which were taken from these rats,were quantitatively analyzed by Western blot and laser confocal microscopy with a double-label immunohistochemical technique. Results Cx43 protein was expressed in both control and USB group, the relative molecular weight was 43000.The mean gray levels of the detrusor protein bands in USB group and control group were 31.066 and 11.701,respectively;the difference of the values between the 2 groups was significant (P
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AIM To get an insight into intracellular signaling steps, a very early step in the signaling cascade, the biphasic Ca 2+ elicited by 5 HT in rat stomach fundus smooth muscle cells was investigated. METHODS Cells were cultured and loaded with Fluo 3 AM. [Ca 2+ ] i was measured by fluorescent intensity (FI) in each cell with confocal microscopy. RESULTS The resting FI level of SFSMC was 264?15. Stimulation of SFSMCs by 5 HT produced an elevation of [Ca 2+ ] i; Depletion of external Ca 2+ by addition of EGTA led to a significant attenuation of [Ca 2+ ] i change induced by 5 HT; Pre treatment of SFSMCs with ryanodine (10 ?mol?L -1 , 5 min) in D Hanks, the effect of 5 HT was completely inhibited; The stimulation of SFSMCs by 5 HT was partly attenuated by miaserin(10 ?mol?L -1 ), however, L type Ca 2+ channel antagonist lacidipine and G protein inhibitor NEM completely abolished the increase of [Ca 2+ ] i mediated by 5 HT; 5 HT mediated Ca 2+ release was reduced by phospholipase C specific inhibitor compound 48/80(1 2 ?g?ml -1 ); When protein kinase C was activated by phorbol 12 myristate 13 acetate (PMA 0 1 ?mol?L -1 , 5 min) the effect of 5 HT was inhibited, and the inhibitory effect of PMA was reversed by D sphingosine, a PKC inhibitor. CONCLUSION Our data suggest that G protein coupled 5 HT 2B receptor in the rat stomach fundus modulates 5 HT stimulated Ca 2+ increase, and it is coupled to calcium influx through L type calcium channels, and also intracellular calcium release by the opening of ryanodine receptor. The 5 HT 2B receptor mediated signal of 5 HT is transduced by PLC and PKC.
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Aim To investigate the effect of iptakalim on neonatal cardiomyocytes hypertrophy induced by isoprenaline(ISO).Methods Hypertrophy in neonatal rat cardiac myocytes was established via culture with ISO.Total protein content was measured by Lowrys method.The fluorescence intensity of intracellular Ca2+ was detected respectively with Fluo-3/AM loading by the laser confocal microscopy.Results ① 1?10-5mol?L-1 ISO can activated ?-AR completely;② the total protein of cardiomyocytes and intracellular i was markedly decreased by treatment with IPT and represented a dose-dependent manner.Conclusion IPT could inhibit the increase of protein content of cardiomyocytes induced by ISO,which may be contributed to the decease of intracellular Ca2+ concentration.