Moderate physical exercise and ATP modulate the P2X7 receptor and improve cisplatin-induced gastric emptying delay in rats

Patients undergoing chemotherapy with cisplatin commonly present gastrointestinal effects such as constipation and gastric emptying (GE) delay. Both the purinergic system and physical exercise modulate the gastrointestinal (GI) tract. In the current study, we investigated the role of ATP, physical exercise, and P2X7 receptor blocking on GE delay induced by cisplatin in rats. Male rats were divided into the following groups: control (C), cisplatin (Cis), exercise (Ex), Brilliant Blue G (BBG), ATP, Cis+Ex, Cis+ATP, Cis+BBG, Cis+Ex+BBG, Cis+Ex+BBG+ATP, and Cis+ATP+BBG. GE delay was induced by treatment with 1 mg/kg cisplatin (1 time/week for 5 weeks, ip). The moderate physical exercise was swimming (1 h/day, 5 days/week for 5 weeks). At the end of the treatment or exercise and 30 min before the GE assessment, some groups received BBG (50 mg/kg, sc) or ATP (2 mg/kg, sc). Then, GE was assessed after a 10-min postprandial period. Chronic use of Cis decreased GE delay (P<0.05) compared to the control group. Both exercise and ATP prevented (P<0.05) GE delay compared to Cis. The pretreatment with BBG significantly inhibited (P<0.05) the effect of exercise and ATP. On the other hand, the association between exercise and ATP reversed (P<0.05) the effect of the BBG and prevented GE delay. Therefore, we suggest that both exercise and treatment with ATP activate P2X7 receptors and prevent GE delay induced by cisplatin in rats.

Up-regulation of P2X7 Receptors Contributes to Spinal Microglial Activation and the Development of Pain Induced by BmK-I / 神经科学通报·英文版

Previous work has demonstrated that the sensitization of spinal neurons and microglia is important in the development of pain behaviors induced by BmK I, a Na channel activator and a major peptide component of the venom of the scorpion Buthus martensi Karsch (BmK). We found that the expression of P2X7 receptors (P2X7Rs) was up-regulated in the ipsilateral spinal dorsal horn after BmK I injection in rats. P2X7R was selectively localized in microglia but not astrocytes or neurons. Similarly, interleukin 1β (IL-1β) was selectively up-regulated in microglia in the spinal dorsal horn after BmK I injection. Intrathecal injection of P2X7R antagonists largely reduced BmK I-induced spontaneous and evoked pain behaviors, and the up-regulation of P2X7R and IL-1β in the spinal cord. These data suggested that the up-regulation of P2X7Rs mediates microglial activation in the spinal dorsal horn, and therefore contributes to the development of BmK I-induced pain.

Terapia fotodinâmica na artrite experimental induzida por Paracoccidioides brasiliensis / Photodynamic therapy in experimental arthritis induced by Paracoccidioides brasiliensis

OBJETIVOS: Avaliar a eficácia da terapia fotodinâmica com Brilliant Blue G no tratamento de um modelo experimental de artrite por Paracoccidioides brasiliensis (P. brasiliensis). MÉTODOS: Após a indução de artrite experimental com isolado de P. brasiliensis da linhagem Pb18 nos joelhos de ratos Wistar, os animais foram divididos em grupos e submetidos a terapia fotodinâmica com fotossensibilizador Brilliant Blue G intra-articular e a laserterapia apenas, sem o Brilliant Blue G. Todos os grupos receberam seus respectivos tratamentos do sétimo ao 11º dia. Para análise do edema foi mensurado o diâmetro latero-lateral do joelho de cada animal diariamente e após o período de tratamento os animais foram sacrificados para dissecação do joelho experimental e coleta de sangue para análise por ELISA, a fim de quantificar os níveis de anticorpos anti-P. brasiliensis. RESULTADOS: A aplicação da terapia fotodinâmica foi capaz de impedir a formação de edema quando comparado ao controle (p>0,005), bem como a produção de anticorpos anti-Gp-43 de P. brasiliensis (p=0,001). No exame anatomopatológico foi possível observar maior grau de sinovite e maior presença de granulomas com o fungo em seu interior no grupo que não recebeu tratamento quando comparado aos grupos que receberam a terapia fotodinâmica. CONCLUSÕES: A terapia fotodinâmica foi eficaz para atenuar a artrite experimental induzida por P. brasiliensis no modelo articular proposto.

AIMS: To evaluate the effectiveness of photodynamic therapy with Brilliant Blue G in the treatment of an experimental model of arthritis by Paracoccidioides brasiliensis (P. brasiliensis). METHODS: After the induction of experimental arthritis with isolated from P. brasiliensis of lineage Pb18 in the knees of Wistar rats, the animals were divided into groups and submitted to photodynamic therapy with intra-articular Brilliant Blue G photosensitizer and laser therapy only, without Brilliant Blue G. All groups received their respective treatments from the seventh to the 11th day. For edema analysis, the knee lateral-lateral diameter of each animal was measured daily and after the treatment period the animals were sacrificed for experimental knee dissection and blood collection for analysis by ELISA, in order to quantify levels of anti-P. brasiliensis antibodies. RESULTS: The results showed that the application of photodynamic therapy was able to prevent the formation of edema when compared to the control (p>0.005), as well as the production of anti-Gp-43 antibodies from P. brasiliensis (p=0.001). In the anatomopathological examination it was possible to observe a higher degree of synovitis and a greater presence of granulomas with the fungus inside the group that did not receive treatment when compared to the groups that received the photodynamic therapy. CONCLUSIONS: Photodynamic therapy was effective in attenuating the experimental arthritis induced by P. brasiliensis in the proposed joint model.

Application of different colorants in the surgery of idiopathic macular hole / 国际眼科杂志(Guoji Yanke Zazhi)

Internal limiting membrane ( ILM) peeling is commonly used in the operation of idiopathic macular hole ( IMH ) surgery. The success of ILM peeling has been greatly improved with the assistance of vital dyes. Currently, several kinds of vital dyes such as indocyanine green ( ICG ) , brilliant blue G ( BBG ) are applied in the ILM staining. However, all of the vital dyes have potential toxicity and side effects on the retina. In recent years, many kinds of dyes and staining improved emerge in endlessly. This paper reports the progress in the application of different colorants in the operation of IMH.

Protective effects of P2X7R antagonist on chronic pancreatitis in mice / 天津医药

Objective To investigate the role of purinergic 2X7 receptor (P2X7R) and its downstream target-NLRP3 inflammasome activation in the process of pancreatic fibrosis in a mouse model of chronic pancreatitis (CP). Methods Forty C57BL/6 mice were randomly divided into normal control group, CP group, P2X7R antagonist oxidized ATP (OxATP) group and brilliant blue G (BBG) group. The chronic pancreatitis model was induced by repeated intraperitoneal injection of the cholecystokinin analogue caerulein with the dose of 50μg/kg for six weeks. Normal saline, OxATP (20μL, 300μmol/L) or BBG (20μL, 10μmol/L) were administered for CP group, OxATP group and BBG group for two weeks after the last caerulein injection. Then all mice were sacrificed and the histopathological changes of the pancreas, especially the fibrotic degrees were evaluated by HE stain, fibrosis score, Sirius red staining and α-SMA immunohistochemical stain. The pancreatic P2X7R, NLRP3 and Caspase-1 expressions were detected by immunohistochemistry respectively to compare the changes between the groups, and explore the role of P2X7R-NLRP3 signaling pathway in pancreatic fibrosis. Results Compared with the normal control group, the scores of pancreatic fibrosis and the expressions of P2X7R, NLRP3 and Caspase-1 in pancreas were significantly increased in CP model group (P<0.05). Compare to CP group, the pancreatic chronic inflammation and the fibrosis indices such as HE fibrosis score, Sirius red staining and α-SMA immunohistochemical stain were ameliorated obviously in OxATP and BBG groups (P<0.05). And expressions of P2X7R, NLRP3 and Caspase-1 in the pancreas were allreduced greatly in both OxATP and BBG groups (P<0.05). Conclusion P2X7R antagonist OxATP and BBG can significantly decrease pancreatic chronic inflammation and fibrosis in the mouse model of CP, which suggests that the blockade of P 2X7R-NLRP3 inflammasome signaling pathway may represent a novel therapeutic strategy for CP and its fibrotic process.

Internal Limiting Membrane Peeling Using 0.025% Brilliant Blue G During Vitrectomy

PURPOSE: To evaluate 0.025% brilliant blue G (BBG) for staining the internal limiting membrane (ILM) during vitrectomy. METHODS: In a retrospective, non-comparative clinical case series, we analyzed consecutive 111 patients who underwent pars plana vitrectomy and removal of the ILM after staining using BBG solution. BBG was dissolved and diluted with balanced salt solution at a concentration of 0.025% and then sterilized by filtering through a 0.22 microm millipore filter. The prepared BBG solution was injected into the vitreous cavity over the macula after removal of the vitreous and excessive solution was removed immediately. RESULTS: The ILM was successfully removed without use of additional adjuvant in all cases. Mean best corrected visual acuity (log MAR) was significantly improved from 0.80 +/- 0.44 at baseline to 0.40 +/- 0.39 at 6 months postoperatively (p < 0.001). One case each of endophthalmitis and diabetic papillopathy developed. The relationship when using BBG solution was not identified as complications were not observed in the other patients who underwent vitrectomy using the same BBG solution on the same day. One idiopathic epiretinal membrane patient had visual acuity loss more than 2 lines. During the follow-up period, other complications suspected to be associated with the use of BBG solution were not observed. CONCLUSIONS: A BBG solution (0.025%) was effective in staining the ILM for removal. Complications associated with the use of BBG solution were not observed.

Internal Limiting Membrane Peeling Using 0.025% Brilliant Blue G During Vitrectomy

PURPOSE: To evaluate 0.025% brilliant blue G (BBG) for staining the internal limiting membrane (ILM) during vitrectomy. METHODS: In a retrospective, non-comparative clinical case series, we analyzed consecutive 111 patients who underwent pars plana vitrectomy and removal of the ILM after staining using BBG solution. BBG was dissolved and diluted with balanced salt solution at a concentration of 0.025% and then sterilized by filtering through a 0.22 microm millipore filter. The prepared BBG solution was injected into the vitreous cavity over the macula after removal of the vitreous and excessive solution was removed immediately. RESULTS: The ILM was successfully removed without use of additional adjuvant in all cases. Mean best corrected visual acuity (log MAR) was significantly improved from 0.80 +/- 0.44 at baseline to 0.40 +/- 0.39 at 6 months postoperatively (p < 0.001). One case each of endophthalmitis and diabetic papillopathy developed. The relationship when using BBG solution was not identified as complications were not observed in the other patients who underwent vitrectomy using the same BBG solution on the same day. One idiopathic epiretinal membrane patient had visual acuity loss more than 2 lines. During the follow-up period, other complications suspected to be associated with the use of BBG solution were not observed. CONCLUSIONS: A BBG solution (0.025%) was effective in staining the ILM for removal. Complications associated with the use of BBG solution were not observed.

Exogenous ATP induces formation of membrane pore in PC12 cells / 中国病理生理杂志

AIM:To investigate the formation of membrane pore in PC 12 cells induced by exogenous adenosine triphosphate ( ATP) and to identify the key molecular targets .METHODS:PC12 cells were treated with different concen-trations of ATP to establish the injury model .The morphological change was observed under an inverted phase -contrast mi-croscope.The viability of the PC12 cells was measured by CCK-8 assay.Fluorescent dye YO-PRO-1 was used to detect the membrane permeability.The expression of P2X7 receptor and pannexin 1 (Panx1) at mRNA and protein levels was as-sessed by real-time PCR and Western blotting .RESULTS:After exposed to ATP (1 mmol/L, 3 mmol/L and 5 mmol/L) for 3 h, the PC12 cells became edematous , and the number of adherent cells decreased gradually in a dose-dependent man-ner .The cell viabilities in 3 mmol/L ATP group and 5 mmol/L ATP group were significantly decreased compared with con-trol group (P0. 05).The expression of P2X7 receptor at mRNA and protein levels was significantly increased (P0.05) when PC12 cells were exposed to ATP for 3 h.CONCLUSION:Extracellular ATP at high concentration may induce membrane pore formation with the expression and activation of P 2X7 receptor in PC12 cells.

A preliminary study of Heavy Brilliant Blue G for internal limiting membrane staining in macular hole surgery.

Context: Surgical outcomes of vitrectomy for idiopathic macular hole using a “heavy” Brilliant Blue G (HBBG) solution for staining and removal of the internal limiting membrane (ILM). Settings and Design: Prospective interventional case series conducted in a tertiary eye care hospital. Materials and Methods: Nineteen patients (20 eyes) with idiopathic macular hole were enrolled to undergo vitrectomy with ILM peeling using HBBG. BBG dye was made heavy by mixing with 10% dextrose normal saline (DNS) solution in 2:1 ratio. The adequacy of ILM staining was noted intraoperatively. The closure rates of macular hole and visual improvement were recorded. Patients were followed up postoperatively on day 1, week 1, and subsequently at 1, 3, and 6 months, and every 6th month thereafter. Statistical Analysis: Wilcoxon signed-rank test was used; P < 0.05 was considered significant. Results: Preoperative best-corrected visual acuity (BCVA) ranged from 20/1000 to 20/63 (median: 20/100). Intraoperatively, the ILM stained very well in all eyes, and was easily removed. All macular holes closed postoperatively. The mean follow-up was 6.15 ± 2 months (range: 4-10; median: 6 months). Final BCVA ranged from 20/20 to 20/80 (median: 20/40), amounting to a significant visual improvement (P = 0.0001). BCVA improved by 1-8 Snellen lines in 19 eyes (95%); 16 eyes (80%) improved by ≥2 lines; 13 eyes (65%) achieved a final BCVA of 20/40 or better. Conclusions: Addition of 10% DNS to BBG dye allowed good ILM staining with less dye during macular hole surgery, and provided excellent anatomic and visual outcomes.

Brilliant blue G-assisted peeling of the internal limiting membrane in macular hole surgery.

Dye-assisted internal limiting membrane (ILM) peeling and gas tamponade is the surgery of choice for idiopathic macular holes. Indocyanine green and trypan blue have been extensively used to stain the ILM. However, the retinal toxicity of indocyanine green and non-uniform staining with trypan blue has necessitated development of newer vital dyes. Brilliant blue G has recently been introduced as one such dye with adequate ILM staining and no reported retinal toxicity. We performed a 23-gauge pars plana vitrectomy with brilliant blue G-assisted ILM peeling in six patients with idiopathic macular holes, to assess the staining characteristics and short-term adverse effects of this dye. Adequate staining assisted in the complete removal of ILM and closure of macular holes in all cases. There was no evidence of intraoperative or postoperative dye-related toxicity. Brilliant blue G appears to be safe dye for ILM staining in macular hole surgery.