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Diabetic retinopathy (DR) is one of the microvascular complications of diabetes mellitus causing severe visual impairment, and it is the main cause of blindness in adults. Metabolic abnormalities play an important role in the occurrence and development of DR, including the abnormal levels of glucose metabolism, lipid metabolism, amino acid metabolism and purine metabolism, which indicate that there are disorders of phosphopentose pathway, arginine metabolism pathway, polyol pathway and ascorbic acid pathway in the progression of DR. Metabolomics has great advantages in exploring the pathogenesis and diagnosis of DR, helping to identify the characteristic metabolic changes of DR And discover potential biomarkers. However, the existing metabolomics studies on DR have some limitations, such as the potential biomarkers found in some studies are difficult to verify in other studies due to differences in race, age, gender and sample size. There are few studies on biomarkers at different stages of DR. Therefore, in the future, multi-center and large-scale clinical studies are needed to screen out biomarkers with practical clinical diagnostic value.
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Objective:To observe any regulatory effect of a pulsed electromagnetic field (PEMF) on A2A adenosine receptors (A2ARs) in the nucleus pulposus of rats with intervertebral disc degeneration (IDD), and to explore any combination with the A2ARs′ agonist-mediating ROS/PI3K/Akt signaling pathway.Methods:Fifty Sprague-Dawley rats were randomly divided into a control group, an intervertebral disc degeneration group (the model group), an A2AR agonist CGS-21680 treatment group (the agonist group), a PEMF group and a PEMF combined with CGS-21680 treatment group (the observation group). IDD was modeled in all except the rats in the control group. 100μL of CGS-21680 (100μg/kg) was injected into the L 5-6 intervertebral discs of the agonist group, while the PEMF group was given 30 minutes of PEMF intervention daily for 14 days at 1.5mT and 75Hz with a pulse width of 150μs. The observation group was injected with CGS-21680 and then given the same PEMF intervention. Primary nucleus pulposus cells from each group (50ng/mL) were cultured and the expressions of 8-OHDG, SOD, MDA and ROS were detected by immunohistochemistry, immunofluorescence or with an ELISA kit. The A2AR, PI3K, AKT and p-AKT protein levels were detected using western blotting. Results:The nucleus pulposus cells and the annulus fibrosus were obviously wrinkled, necrotic and broken in the model group but the annulus fibrosus was intact and the nucleus pulposus was almost normal in the observation group. Compared with the model group, the levels of SOD and A2AR, PI3K, p-AKT and AKT protein were higher in the agonist, PEMF and observation groups, while the expressions of MDA, ROS and 8-OHDG were weaker. The ROS level in the observation group was significantly lower than that in the agonist and PEMF groups, and the phosphorylation level of p-AKT in the observation group was significantly higher than in the agonist and PEMF groups. The average levels of SOD, A2AR, PI3K, p-AKT and AKT protein in the nucleus pulposus cells of the agonist, PEMF and observation groups were significantly higher than the IL-1β group′s average, while the average levels of MDA, ROS and 8-OHDG were significantly lower. The ROS levels in the observation group were significantly lower than in the agonist and PEMF groups, while the A2AR protein content and p-AKT phosphorylation in the observation group were significantly greater. The average Bax levels in the nucleus pulposus cells of the agonist, PEMF and observation groups were significantly lower than that in the IL-1β group, and the expression of Bcl-2 was significantly increased. There was significantly less apoptosis observed in the observation group than in the agonist and PEMF groups, while the expression of Bcl-2 was significantly higher.Conclusions:PEMF plays an anti-oxidative stress role by up-regulating A2AR activity and reducing ROS generation. Treatment with PEMF and A2AR agonist could further activate the phosphorylation of PI3K/Akt, down-regulate Bax and up-regulate Bcl-2, thus inhibiting the apoptosis of nucleus pulposus cells and alleviating the malignant progression of IDD.
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Objective:To explore the effect of combining pulsed electromagnetic field (PEMF) stimulation with A 2A adenosine receptor agonist CGS-21680 on apoptosis and inflammation of nucleus pulposus cells in cases of intervertebral disc degeneration. Methods:Forty-eight Sprague-Dawley rats were randomly divided into a sham operation group (Sham group), an intervertebral disc degenerative disease group (Model group), an A 2A adenosine receptor agonist CGS-21680 group (Agonist group), and a group in which PEMF was combined with CGS-21680 (Observation group). The rat model of intervertebral disc degeneration (IDD) was established in all other groups than the sham operation group. The rats in the Agonist group were injected with 100μL of CGS-21680 (100μg/kg) at the L 5-6 intervertebral disc. The Observation group was injected with CGS-21680 similarly but then received 14 conse-cutive days of PEMF stimulation (30min/time). The Sham and Model groups were injected with the same amount of normal saline solution. Eight weeks later, HE staining was used to evaluate the pathological changes in the intervertebral disc tissues. The expression of type II collagen was determined by immunohistochemistry. The content and mRNA expression of inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were determined using enzyme-linked immunosorbent assays and reverse transcription-polymerase chain reactions (RT-PCRs). The protein and mRNA levels of A 2A, NLRP3 and caspase-3 were determined by western blotting and RT-PCR. Results:The degeneration in the nucleus pulposus and annulus fibrosus in the Model group was significant, while significantly less shrinkage, necrosis and fibrous annulus rupture was observed in the Observation group. Compared with Model and Agonist groups, the positive expression of Col Ⅱ in the nucleus pulposus, A 2AR protein levels and relative expression of its mRNA had all increased significantly in the Observation group, while the average levels and mRNA expression of pro-inflammatory cytokines IL-6 and TNF-α had decreased significantly. The average protein level and mRNA expression of NLRP3 and caspase-3 in the intervertebral disc tissues of the Observation group were significantly lower than in the Model and Agonist groups. Conclusions:Combining pulsed electromagnetic field stimulation with A 2A adenosine receptor agonist CGS-21680 can inhibit the apoptosis of nucleus pulposus cells, alleviate disease response and delay IDD by up-regulating the activity of A 2A receptors and down-regulating the expressions of pro-inflammatory factors IL-6, TNF-α, NLRP3 and caspase-3 in nucleus pulposus cells.
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Objective:To observe electromagnetic field irradiation′s ability to inhibit heterotopic ossification (HO) and explore the mechanisms involved.Methods:A total of 72 male Sprague-Dawley rats were randomly divided into a control group, a model group and an observation group, each of 24. The rats in the model and observation groups were made to model HO by clamping and cutting the Achilles tendon, while among those in the control group the Achilles tendon was only exposed without clamping and cutting. The rats in the observation group were exposed to a 1mT electromagnetic field alternating sinusoidally at 50Hz (SEMF) twice a day for 2h each time. At 3 days, 14 days, 1 month and 3 months after the operation, Achilles tendon tissue was resected from 6 rats in each group for gross and histological examination and immunohistochemical staining. The protein content and gene expression were detected using western blotting or reverse transcription polymerase chain reaction. X-ray examination was performed at 1 and 3 months after the operation to observe any HO.Results:No significant changes were observed in the control group after the operation. Compared with the model group, HO in the observation group was significantly inhibited. The average expression of the MMP-9 gene in the model and observation groups was significantly higher than in the control group, and it increased with time. The average tissue metalloproteinase-1 (TIMP-1) gene expression was significantly lower, and it decreased with time. Compared with the model group, the average expression of MMP-9 in the observation group was significantly less, while that of TIMP-1 was significantly greater. Compared with the model group, there was also a significant increase in the expression of decapentaplegic homolog 7 (Smad7) protein and a significant decrease in the expression of phosporylated Smad3 protein at each time point.Conclusion:Applying 50Hz, 1mT SEMF has a definite effect in inhibiting HO, at least in rats. It may be related to promoting the expression of Smad7 and inhibiting the phosphorylation of Smad3, thus inhibiting the TGF-β/Smad signal transduction pathway and promoting the restoration of a dynamic balance of MMP-9 with TIMP-1.
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Objective@#To explore the expression of the A2A adenosine receptor and the inflammatory cytokines interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) in human degenerative nucleus pulposus (NP) cells after they have been treated with a pulsed electromagnetic field (PEMF).@*Methods@#Human degenerative NP cells were cultured in vitro and treated using an 0.8mT PEMF with a pulse frequency of 50Hz. The pulse width was 150μs and the exposure time was 30min, repeated 5 times at 12 hour intervals. The expression of the A2A adenosine receptor in NP cells was determined using western blotting and reverse transcription polymerase chain reactions. The expression of the inflammatory cytokines IL-1β and TNF-α were detected using enzyme-linked immunosorbent assays (ELISA). The human degenerative NP cells were also treated with an antagonist and agonist of the A2A adenosine receptor, and the expression of IL-1β and TNF-α were also determined using ELISA.@*Results@#After the PEMF treatment the expression of the A2A adenosine receptor increased significantly, while the expression of IL-1β and TNF-α decreased significantly. However, the A2A adenosine receptor antagonist reversed the inhibitory effect of the PEMF on the expression of IL-1β and TNF-α, while the agonist played an opposite role.@*Conclusion@#A PEMF can significantly inhibit the expression of IL-1β and TNF-α in human degenerative NP cells, which could be related to up-regulation of the expression of the A2A adenosine receptor in those cells.
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Objective@#To explore the epidemiological distribution characteristics of glucose-6-phosphate dehydrogenase (G6PD) activity, incidence of G6PD deficiency in neonates and the cut-off values. @*Methods@#About 1.44 million newborns in 10 districts of Zhejiang province from March 2015 to September 2017 were included in this study. Fluorescence analysis was used to determine the G6PD activity in dried blood spots. Those with initial screening positive results were recalled and confirmed by direct ratio of G6PD to 6PGD (6-phosphogluconate dehydrogenase) to confirm the diagnosis. The results were analyzed by using nonparametric and chi-square tests. @*Results@#Significant differences of G6PD levels were found among the groups of different genders, gestational age, birth weight, blood sampling age, blood sampling season and districts (P＜0.01). The male incidence of G6PD deficiency was significantly higher than female incidence. In different regions of Zhejiang province, the highest prevalence was in Lishui (0.38%) and the lowest was in Zhoushan (0.11%), The trend of high prevalence in the south and low prevalence in the north was basically showed. When the cut-off value of G6PD activity ranged from 2.60 to 2.80 U/g Hb, the sensitivity of G6PD deficiency screening for male and female newborns was 100% and the Youden index was the highest (about 0.99). @*Conclusion@#The level of G6PD activity may be relevant to the factors of population group and period. The incidence of G6PD deficiency may be affected by different genders and different regions. The cut-off values for screening may initially set at 2.60 U/g Hb and 2.80 U/g Hb for male and female respectively.
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Objective To evaluate the left ventricle geometry shape and left heart regional function in mitral insufficiency (MI) by quantitative tissue velocity imaging (QTVI) and the left ventricle systolic and diastolic geometry spbericity index. Methods Thirty normal subjects and 62 MI patients underwent QTVI and color Doppler imaging (CDFI) were enrolled in this study in order to measure the left ven-tricular geometry shape and left ventrieular regional function along LV apical long-axis view. Off-line LV regional velocity images along LV apical long-axis view were synchronously obtained. Peak tissue velocities of LV regional muscular tissue during systole(Vs),systolic acceler-ation(a), early diastole(Ve), LA contraction(Va)were synchronously measured as the index of left ventricular regional systolic and dias-tolic function. The left ventricle geometry shape index were reflected from the systolic and diastolic geometry sphericity index (SIs and Sid) and the left ventrieular ejection fraction (LVEF) and the peak D and peak a (PVd/PVa) of pulmonary veins flowing spectrum reflected the global left ventricular function index. The Vs, Ve, Va, a, PVd/PVd ratio, LVEF, SIs, SId were recorded and their correlation between normal subjects and patients with MI were compared. Result Vs, Ve,Va,a,PVd/ PVa ratio,SIs,SId in patients with MI were significantly re-duced. There was positive relation between Sis and a(r=0.602)and Ve/Va and SId(r=0.635). Conclusion There was negative rela-tion between regional cardiac function and LV spberieity, the higher cardiac function was accompanied with the lower SI.
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BACKGROUND: In clinical practice, delayed brain injury after carbon monoxide (CO) poisoning occurs in 3% to 30% of the persons who suffered carbon monoxide poisoning and is in the main presence of dementia, psychiatric symptom and extrapyramidal symptoms. At present, its pathogenesis is unclear.OBJECTIVE: To investigate the pathological damage mechanism of delayed brain injury after CO poisoning and the protective effect of hyperbaric oxygen on delayed brain injury.DESIGN: A randomized controlled animal experiment.SETTING: Staff Room of Aviation Health, Department of Aerospace Medicine, the Fourth Military Medical University of Chinese PLA.MATERIALS: This experiment was carried out in the Laboratory of Aviation Pathology and Molecular Biology, Department of Aerospace Medicine, the Fourth Military Medical University of Chinese PLA in March 2004. Totally 80 healthy male Sprague-Dawley, of clean grade, were used in this experiment. The animals were randomized into 3 groups: normal control group (n=10), model group (n=35) and hyperbaric oxygen group (n=35).The latter two groups were separately divided into 7 time points: poisoning 6 hours, 1,3,5,7,14 and 21 days, 5 rats at each time point.METHODS: In the model group, acute CO poisoning rat models were developed by placing the rats in the poisoning jar which contained the mixture of CO and air for 60 minutes. The volume faction of CO was kept at 2 500×10-6. In the hyperbaric oxygen group, modeling was the same as that in the model group. Three hours after poisoning, the rats were given 115-minute hyperbaric oxygen treatment. Pressure was 0.2 Mpa and the volume fraction of oxygen was over 0.90. The first three days after poisoning, hyperbaric oxygen treatment was conducted twice per day, then once per day, with one day of non-administration in a week. There was no intervention in the normal control group. MAIN OUTCOME MEASURES: ① The characters of pathological changes in brain tissue of rats at each time point after poisoning were detected with histopathological and immunohistochemical methods; ② Neuronal apoptosis was detected with electron microscopy and in situ TdT-mediated-dUTP nick end labeling(TUNEL).RESULTS: After rats were modeled, the rate of death was about 10%. ① In the model group, general pathological injury occurred in the brains of rats. Denatured necrosis appeared in the neurons of cerebral cortex, hippocampus, corpus striatum, cerebellum and other regions. Injuries in cerebral cortex, hippocampus and other regions were severe. Results of haematoxylin-eosin and TUNEL staining and electron microscope observation demonstrated that apoptosis occurred in hippocampal neurons. The apop totic neurons increased on the 3rd day after poisoning, reached the peak on the 7th day (P ＜ 0.01), then gradually decreased. In the hyperbaric oxygen group, the denatured necrosis of neurons in the brains was significantly lightened, and the injuries of hippocampal region of rats at each time point were significantly attenuated in comparison with model group; The number of necrotic neurons was decreased, especially on the 5th and 7th days after poisoning (P ＜ 0.01). Hyperbaric oxygen promoted the expression of Bcl-2 in hippocampus of modeled rats, especially on the 3rd and 5th days after Coexposure (P ＜ 0.01).CONCLUSION: General delayed neuronal injury is found in the acute CO poisoning rat, with the presence of delayed neuronal necrosis and apoptosis. Hyperbaric oxygen treatment can effectively reduce denatured and necrotic neurons and promote the expression of apoptosis-inhibiting gene bcl-2, then inhibit neuronal necrosis and apoptosis.
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[Objective] To study the effect and reliability of fluoxetine on depression and symptoms in cancer patient,and to assess the overall quality of life before and after treatment.[Methods]We treated 54 cancer patients with depression by Fluxetine for 8 w,at the same time,evaluated their emotion state,change of quality of life and the adverse effect with HAMD,HAMA,TESS and laboratory tests.[Results]After 8 w,patients’ scores of anxiety and depression decreased significantly from the baseline,effective rates were 82.4% and 96.3% respectively.3 domains of quality of life (physiology,psychology and independence) became much better than those of baseline.The side effect of fluxetine was small.[Conclusion]This study shows that fluoxetine can reduce the depression and anxiety symptoms in cancer patients.
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BACKGROUND: Hyperbaric oxygen(HBO) is the first choice in the treatment of acute carbon monoxide(CO) poisoning. However,the mechanism of HBO in the treatment of CO poisoning,especially the mechanism in the treatmentof CO poisoning-induced delayed encephalopathy,is unclear at present.OBJECTIVE: To observe the pathological changes of hippocampal neurons in rats after acute CO poisoning to investigate the impacts of HBO therapy on Bcl-2 protein expression in hippocampal neurons in rats after CO poisoning.DESIGN: A completely randomized controlled experimental study based on the experimental animals.SETTING: Emergency department in a military medical university of Chinese PLA affiliated hospital,department of laboratory medicine in a municipal hospital,and the center of HBO therapy in a military medical university of Chinese PLA.MATERIALS: The study was conducted in the Laboratory of HBO Therapy Center,Faculty of Aerospace Medicine,the Fourth Military Medical University of Chinese PLA. Sixty male SD rats were selected.INTERVENTIONS: Sixty rats were randomly divided into 3 groups: normal control group(control group),CO poisoning group(CO group),and HBO therapy group(HBO group) with 20 rats each. Rats of each group were exposed under air or CO gas(volume fraction was 3.2 × 10-3) respectively for 60 minutes,and rats of CO-HBO group were treated by HBO. Cerebral pathological slices of hippocampus were prepared for routine HE and Bcl-2 staining to observe the characteristics of the changes of hippocampal neuronal injury and the Bcl-2protein expression on the 1st,3rd,5th and 7th day after CO poisoning.MAIN OUTCOME MEASURES: Changes of pathomorphology and Bcl-2protein expressionRESULTS: It could be seen lot of degenerated and necrotic neurons in hippocampus of rats in CO group. Degenerated and necrotic neurons decreased and the expression of Bcl-2 protein increased in CO-HBO group,especially on the 3rd and 5th day after poisoning( P ＜ 0. 05).CONCLUSION: HBO therapy can promote Bcl-2 protein expression in hippocampus after acute CO poisoning,so it can protect neurons.