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Objective To investigate the neuroprotective effect of methylene blue on diabetic retinopathy in rats. Methods Thirty SD rats were randomly divided into blank, control and experimental groups. The control and experimental groups were induced with diabetes by streptozotocin (STZ) intraperitoneal injection. After 6 weeks of successful modeling, the experimental group received intravitreal injection of methylene blue at a dose of [0.2 mg/(kg.d)], while the control group received an equal amount of dimethyl sulfoxide (DMSO) intravitreal injection, both continuously injected for 7 days. ELISA was used to detect the levels of retinal superoxide dismutase (SOD), 8-iso-prostaglandin F2alpha (iPF2α) and interleukin-1β (IL-1β) in rats. Western blot analysis was used to detect the expression of retinal extracellular signal-regulated kinase 1/2 phosphorylation (p-ERK1/2) and phosphorylated protein kinase B (p-AKT), and PAS staining was used to detect retinal morphological changes. Results Compared with the blank group rats, the retinal SOD activity in the control and experimental group rats was significantly reduced. iPF2α, IL-1β and p-ERK1/2 level increased, while p-AKT level decreased. Compared with the control group, the SOD activity of the experimental group rats increased. iPF2α and IL-1β level went down, while p-ERK1/2 and p-AKT level went up significantly. The overall thickness of the retinal layer and the number of retinal ganglion cells were significantly reduced. Conclusion Methylene blue improves diabetic retinopathy in rats by reducing retinal oxidative stress and enhancing ERK1/2 and AKT phosphorylation.
Subject(s)
Rats , Animals , Diabetic Retinopathy/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , Interleukin-1beta/metabolism , Methylene Blue/pharmacology , Phosphorylation , Rats, Sprague-Dawley , MAP Kinase Signaling System , Diabetes Mellitus, Experimental/drug therapy , Superoxide Dismutase/metabolismABSTRACT
Migraine is a common neurological disease.It involves complex neurological abnormalities.Recent advances in the neurophysiology of migraine have enabled us to explain some of the symptomatic problems and have contributed to the development of new targeted treatments that may change the way migraine treated in the future.Migraine treatment is individualized,in which preventive drug therapy also plays an important role.This article will discuss the new progress in the treatment of migraine,with emphasis on the new treatment of calcitonin gene-related peptide pathway.
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OBJECTIVE： To observe the protective effect of atorvastatin-induced increase of EPC-MVs on myocardial cells in ST-segment elevation myocardial infarction （STEMI） patients， and to investigate its potential mechanism. METHODS： Totally 168 STEMI patients was collected from our hospital during Feb. 2015-Feb. 2018， and then divided into group A （88 cases） and group B （94 cases） according to the dose of atorvastatin. All patients received percutaneous coronary intervention， and then given Bivaleridine for injection， Clopidogrel bisulfate tablets and Atorvastatin calcium tablets. Group A was given Atorvastatin calcium tablets 20 mg， once a day. Group B was given Atorvastatin calcium tablets 20 mg， twice a day. A treatment course lasted for 30 d， and two groups were treated for 3 courses at least. The levels of blood lipid （TC， LDL-C， HDL-C） （before treatment and 30th， 60th， 90th day after treatment） and the number of EPCs positive cells （30th， 60th day after treatment） were observed in 2 groups. The expression of miRNA of EPC-MVs （60th day after treatment） was detected， and the expression difference of miRNA were validated. Target gene and KEGG pathway enrichment of miRNA with most significant expression difference were analyzed， and the effects of it on the proliferation of cardiac HCM-a cells were evaluated. The occurrence of ADR was recorded in 2 groups. RESULTS： Totally 8 patients withdrew from the study in group A， and 6 patients in group B. There was no statistical significance in the levels of TC， LDL-C and HDL-C or the number of EPCs positive cells in peripheral blood between 2 groups before treatment or 30th day after treatment （P＞0.05）. After treatment， the level of HDL-C in 2 group （60th and 90th day after treatment） and the number of EPCs positive cells in peripheral blood in group B （60th day after treatment） were increased significantly， and group B was significantly higher or more than group A at the same time point （P＜0.05）. Microarray analysis showed that compared with group A， 16 miRNAs expressed more than 1.5 times differentially in EPC-MVs of group B， 7 of which were up-regulated and 9 down-regulated. Top five differentially expressed genes were hsa-miR-126 （up-regulated）， hsa-miR-1275 （up-regulated）， hsa-miR-7704 （down-regulated）， hsa-miR-105-5p （down-regulated）， and hsa-miR-3180 （down-regulated）. Fluorescence quantitative polymerase chain reaction results showed that compared with group A， relative expression of hsa-miR-126 and hsa-miR-1275 in group B were increased significantly； and relative expression of hsa-miR-7704， hsa-miR-105-5p and hsa-miR-3108 were decreased significantly （P＜0.05）. The expression difference of hsa-miR-126 was the most significant， and its target genes included Ang-1， PDGF， p38 MAPK， Smad2/3， HIF-1， TGF-β， etc. The signaling pathways involved in regulation mainly included angiogenesis signaling pathway， chronic myelogenous leukemia related pathway， renal epithelial cell carcinoma related pathway and so on. CCK-8 test showed that the optical density （OD） of cells in hsa-miR-126 specific interfering substance group was decreased significantly， and the OD value of cells in simulated substance group was increased significantly， compared with blank group （P＜0.05）. There was no statistical significance in the incidence of ADR as diarrhea， nausea and vomiting， rash， etc. （P＞0.05）. CONCLUSIONS： Different doses of atorvastatin can regulate the level of HDL-C， and large dose of atorvastatin can increase the number of EPCs significantly， but dose not influence the safety of drug use. This effect may be associated with up-regulating the expression of hsa-miR-126 in EPC-MVs so as to promoting the proliferation of myocardial cells.
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Forty three medical graduates from Hainan Medical University participating in the standardized general practice residency training in 2015 and 2016 were randomly divided into study group (n=22) and control group (n=21). The study group received a mid-term assessment and formative assessment, and the control group received the final assessment only. At the end of the six months of training, the scores of the mid-term assessment in study group were significantly higher than those in the control group [(82.9±8.4) vs. (75.5±10.2), t=12.706, P<0.01]. A questionnaire survey was conducted among the participants in study group, 86.4% of them recognized and supported the formative assessments. Formative assessment can improve the teaching quality of general practice residency training. The trainees have a positive attitude towards this assessment mode.
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Stem cells transfer normal functional mitochondria to injured lung cells and reconstruct their function, which is one of the important mechanisms of stem cells in the treatment of lung injury. Mitochondrial translocation is mediated by intercellular gap junction (GJCs), which is unidirectional. This paper reviewed the research of stem cells for the treatment of lung injury mitochondrial transfer mechanism, to understand the clinical significance of lung mitochondria in physiological and pathological roles of mitochondria and transfer mechanisms of stem cells for the treatment of various types of lung injury.
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Objective To summarize the nursing experience of 1 case of very low birth weight infant decannulation difficult in PICC. Methods The nursing key points included: full assessment analysis decannulation difficult reason, consult the PICC catheter outpatient health nurses, give magnesium sulfate hydropathic compress, mucopolysaccharide polysulfate cream local besmear outside, sanyrene outside, at the same time give low-molecular-weight heparin calcium injection subcutaneous injection such as anticoagulant active treatment and nursing. Results After 7 days ,the infant left axillary mass dispel, PICC pull out smoothly. Conclusions Decannulation difficult of very low birth weight infant requires full evaluation, multidisciplinary cooperation and specialist consultation, can give targeted personalized nursing safety smooth tube drawing, is worthy of reference for clinic.
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Objective:To prepare resveratrol microemulsions by high pressure homogenization method and characterize the microe-mulsions. Methods:Using the particle size, polydispersion index and encapsulation efficiency as the indicators, the independent varia-bles of the preparation were inspected, and the microemulsions were characterized. The stability of resveratrol microemulsions was stud-ied by long term stability test preliminarily. Results:The mean particle size, polydispersion index and zeta potential of resveratrol mi-croemulsions was (231 ± 37. 8) nm, 0. 228 ± 0. 047 and ( -42. 5 ± 4. 3) mV, respectively. The microemulsions were found to be small and spherical with smooth surface under a transmission electron microscope. Long term stability studies showed that the microe-mulsions were stable in 3 months after stored at 25℃. Conclusion:The preparation process of high pressure homogenization method for resveratrol microemulsions is simple and feasible.
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With the development of the research ,gastrin secrete element and Reg gene families ,especial-ly the RegⅠand RegⅣ have a close relationship in biological behavior ,development and progress of digestive system tumor,the latter maybe play a role of oncogenes .So combination of gastrin with REG gene may contribute to the diagnosis of early gastric cancer ,and provides new target for prevention of gastric cancer .The present review summarized the related research progress on expression as well as clinical significant of gastrin and Reg gene in gastric cancer tissue .
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bThis study explored whether the transplantation of modified marrow stromal cells (MSCs) has angiogenic effects in a left middle cerebral artery occlusion infarction/reperfusion (MCAO I/R) rat model and preliminarily examined the mechanism of angiogenesis following cerebral infarction. MSCs were isolated by using a direct adherent method and cultured. Vascular endothelial growth factor (VEGF) was transfected into MSCs by employing the liposome transfection. The transfection efficiency was measured by the optical density method. The protein expression of VEGF gene before and after transfection was measured by Western blotting. SD rat model of transient occlusion of the left middle cerebral artery was established by using an approach of intra-luminal occlusion. Tetrazolium (TTC) and HE staining were performed to observe the cerebral infarction. ELISAs were used to measure the levels of VEGF in the rat cerebral tissues. The expression patterns of angiopoietin-2 (Ang-2) and CD34 in cells surrounding the area of infarction were immunohistochemistrically observed. Ang-2 protein expression in the tissue surrounding the area of infarction was measured by Western blotting. VEGF expression in the MSCs increased after transfection at a rate of approximately 28%±3.4%. ELISA showed that the expression of VEGF in the cerebral tissue was significantly increased after induction of infarction, peaking on the 4th day and decreasing to the levels of the sham surgery group (normal) within 7 to 10 days. The VEGF level was significantly higher at each time point in the VEGF-MSC and MSC groups compared to the model group. Moreover, the VEGF level was higher in the VEGF-MSC group than in the MSC group and stayed relatively high until the 10th day. The immunohistochemical results showed that 10 days after the infarction, the number of Ang-2 and CD34-expressing cells in the area surrounding the infarction was significantly higher in the VEGF-MSC group and the MSC group compared to the model group. Moreover, the VEGF level was higher in the VEGF-MSC group than the MSC group. A similar trend in Ang-2 protein expression was revealed by Western blotting. In the MCAO rat model transfected with modified MSCs over-expressing VEGF, compared to the MSC transplantation group, the concentration of VEGF was significantly increased in the brain tissue after cerebral infarction. In addition, the level of Ang-2 was up-regulated, with angiogenesis promoted, the blood supply to the areas surrounding the cerebral infarction increased, and neurological function improved. We are led to speculate that the synergistic effects of VEGF and Ang-2 may be responsible for the angiogenesis following cerebral infarction.
Subject(s)
Animals , Male , Rats , Angiopoietin-2 , Genetics , Metabolism , Bone Marrow , Metabolism , Pathology , Cerebral Infarction , Genetics , Metabolism , Pathology , Neovascularization, Pathologic , Genetics , Pathology , Rats, Sprague-Dawley , Stromal Cells , Metabolism , Pathology , Vascular Endothelial Growth Factor A , Genetics , MetabolismABSTRACT
This study examined the effect of electro-acupuncture (EA) combined with transcranial magnetic stimulation (TMS) therapy at different time windows on learning and memory ability of rats with cerebral infarction and the underlying mechanism. Two hundred SD rats were randomly divided into four groups: normal group, sham-operated group, model group and EA+TMS group, and each group was then divided into five sub-groups in terms of the different time to start treatment post operation: 6, 12, 24, 48 and 72 h. Cerebral infarction models were established in the model and the EA+TMS groups by left middle cerebral artery occlusion/reperfusion (MCAO/R). After treatment for 14 d, the Morris water maze test was applied to examine the spatial learning and memory abilities of rats. In infarcted area, the expression of caspase-3 was immunohistochemically detected, and real-time fluorescent quantitative PCR was used to measure the expression of Bcl-2 mRNA. The results showed that in EA+TMS group compared with model group at the same treatment time windows, the escape latency was substantially shortened, the expression of caspase-3 was considerably decreased and the expression level of Bcl-2 mRNA significantly increased (P<0.05). In the EA+TMS sub-groups, the escape latency was shortest, the expression level of caspase-3 lowest, and the expression level of Bcl-2 mRNA highest at the treatment time window of 24 h. It was concluded that EA combined with TMS can promote neurological function of rats with cerebral infarction by increasing the expression level of Bcl-2 mRNA and decreasing the expression of caspase-3. The best time window is 24 h after perfusion treatment to ischemia.
Subject(s)
Animals , Rats , Acupuncture Therapy , Methods , Apoptosis , Physiology , Cerebral Infarction , Learning , Physiology , Memory , Physiology , Neurons , Physiology , Rats, Sprague-Dawley , Transcranial Magnetic Stimulation , MethodsABSTRACT
This study examined the effect of electro-acupuncture (EA) combined with transcranial magnetic stimulation (TMS) therapy at different time windows on learning and memory ability of rats with cerebral infarction and the underlying mechanism. Two hundred SD rats were randomly divided into four groups: normal group, sham-operated group, model group and EA+TMS group, and each group was then divided into five sub-groups in terms of the different time to start treatment post operation: 6, 12, 24, 48 and 72 h. Cerebral infarction models were established in the model and the EA+TMS groups by left middle cerebral artery occlusion/reperfusion (MCAO/R). After treatment for 14 d, the Morris water maze test was applied to examine the spatial learning and memory abilities of rats. In infarcted area, the expression of caspase-3 was immunohistochemically detected, and real-time fluorescent quantitative PCR was used to measure the expression of Bcl-2 mRNA. The results showed that in EA+TMS group compared with model group at the same treatment time windows, the escape latency was substantially shortened, the expression of caspase-3 was considerably decreased and the expression level of Bcl-2 mRNA significantly increased (P<0.05). In the EA+TMS sub-groups, the escape latency was shortest, the expression level of caspase-3 lowest, and the expression level of Bcl-2 mRNA highest at the treatment time window of 24 h. It was concluded that EA combined with TMS can promote neurological function of rats with cerebral infarction by increasing the expression level of Bcl-2 mRNA and decreasing the expression of caspase-3. The best time window is 24 h after perfusion treatment to ischemia.
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bThis study explored whether the transplantation of modified marrow stromal cells (MSCs) has angiogenic effects in a left middle cerebral artery occlusion infarction/reperfusion (MCAO I/R) rat model and preliminarily examined the mechanism of angiogenesis following cerebral infarction. MSCs were isolated by using a direct adherent method and cultured. Vascular endothelial growth factor (VEGF) was transfected into MSCs by employing the liposome transfection. The transfection efficiency was measured by the optical density method. The protein expression of VEGF gene before and after transfection was measured by Western blotting. SD rat model of transient occlusion of the left middle cerebral artery was established by using an approach of intra-luminal occlusion. Tetrazolium (TTC) and HE staining were performed to observe the cerebral infarction. ELISAs were used to measure the levels of VEGF in the rat cerebral tissues. The expression patterns of angiopoietin-2 (Ang-2) and CD34 in cells surrounding the area of infarction were immunohistochemistrically observed. Ang-2 protein expression in the tissue surrounding the area of infarction was measured by Western blotting. VEGF expression in the MSCs increased after transfection at a rate of approximately 28%±3.4%. ELISA showed that the expression of VEGF in the cerebral tissue was significantly increased after induction of infarction, peaking on the 4th day and decreasing to the levels of the sham surgery group (normal) within 7 to 10 days. The VEGF level was significantly higher at each time point in the VEGF-MSC and MSC groups compared to the model group. Moreover, the VEGF level was higher in the VEGF-MSC group than in the MSC group and stayed relatively high until the 10th day. The immunohistochemical results showed that 10 days after the infarction, the number of Ang-2 and CD34-expressing cells in the area surrounding the infarction was significantly higher in the VEGF-MSC group and the MSC group compared to the model group. Moreover, the VEGF level was higher in the VEGF-MSC group than the MSC group. A similar trend in Ang-2 protein expression was revealed by Western blotting. In the MCAO rat model transfected with modified MSCs over-expressing VEGF, compared to the MSC transplantation group, the concentration of VEGF was significantly increased in the brain tissue after cerebral infarction. In addition, the level of Ang-2 was up-regulated, with angiogenesis promoted, the blood supply to the areas surrounding the cerebral infarction increased, and neurological function improved. We are led to speculate that the synergistic effects of VEGF and Ang-2 may be responsible for the angiogenesis following cerebral infarction.
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Objective To study the effect of electroacupuncture (EA) combined with transcranial magnetic stimulation (TMS) on the expression of the B-cell lymphoma/leukemia-2 gene (Bcl-2) and brain derived neurotrophic factor (BDNF) after cerebral infarction. Methods One hundred Sprague-Dawley rats were randomly and equally divided into a normal group, a sham-operated control group, a model group and an EA plus TMS group. A cerebral infarction model was established in the latter two groups using left middle cerebral artery occlusion (MCAO). Five-member subgroups of the EA plus TMS group were then treated at 6, 12, 24,48 and 72 hours after reperfusion. Sham EA plus TMS was given to similar sub-groups from the other groups at the same time points. The expression of Bcl-2 mRNA and BDNF mRNA were measured using a RT-PCR at the 14th day. Results Positive expression of Bcl-2 mRNA and BDNF mRNA was detected around the infarction in all groups. The average expression of both was significantly higher in the EA plus TMS group than in the model group. Bcl-2 mRNA peaked when the therapy was administered at 24 hours and BDNF mRNA at 48 hours.Conclusions The expression of Bcl-2 mRNA and BDNF mRNA is maximized when EA plus TMS is administered 24-48 hours after cerebral infarction. EA plus TMS does have protective and rehabilitative effects on rats after cerebral infarction.
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Objective To study the effect of transcranial magnetic stimulation (TMS) on the rehabilitation of rats with cerebral infarction. Methods One hundred Sprague-Dawley rats were randomly divided into a normal group, a sham-operated control group, a model group and a TMS group with 25 rats in each group. A cerebral infarction model was established in the latter two groups by left middle cerebral artery occlusion (MCAO). TMS was started at either 12 or 24 hours after reperfusion, and sham-TMS was given to the first two groups at the same time points. The expression of Bcl-2 mRNA and BDNF mRNA were measured by RT-PCR after 14 days. Results Bcl-2 mRNA and BDNF mRNA were detected in all groups. The expression of Bcl-2 mRNA in the TMS-12 h group, and that of BDNF mRNA in the TMS-24 h group were significantly higher than in the other groups. Conclusions The expression of Bcl-2 mRNA and BDNF mRNA in the brains of rats after cerebral infarction peak when TMS is administered 12 h and 24 h after reperfusion, respectively. TMS might have protective and rehabilitative effects on rats after cere-bral infarct.
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Objective To investigate the changes of HPA axis activity following scald stress and to elucidate if NMDA receptor is involved in this change. Methods Adult male Wistar rats were inflicted with 30% TBSA full thickness scalding burn, which was applied as severe trauma stress model. Using this model, we detected the changes of serum cortisol and ACTH concentration in scald rats pretreated with intrahippocampal microinjection of NMDA receptor antagonist MK801 or NMDA receptor 1 antisense oligodeoxynucleotide. Results Intrahippocampal microinjection of MK 801 6 ?g resulted in an significant decrease of serum cortisol and ACTH concentration at 2 h after burn, and microinjection of MK 801 12 ?g resulted in a more significant decrease of these values. In accordance with microinjection of MK 801, intrahippocampal microinjection of NMDA receptor 1 antisense oligodeoxynucleotide of 10 nmol/L and 20 nmol/L also resulted in a significant and a more significant decrease of serum cortisol and ACTH concentration after burn. Conclusion Hippocampal NMDA receptor plays an important role in over excitation of HPA axis following burn.