Objective:To document any effect of a pulsed electromagnetic field (PEMF) on the regulation of neuropeptide Y (NPY) in nucleus pulposus (NP) tissue, NP cell apoptosis and matrix degradation using rats with intervertebral disc degeneration (IDD).Methods:Eighteen Sprague-Dawley rats were randomly divided into a control group, an IDD model group (the model group), and a PEMF group. IDD was induced in both the model and PEMF groups. Right after the modeling, the PEMF group received 14 days of PEMF treatment, while the control group and model group were given no special treatment. Meanwhile, the primary rat nucleus pulposus cells (NPCs) were cultured using Dulbecco′s Modified Eagle Medium at 37℃ and 5% CO 2. When the fusion rate reached 90% after passage, the NPCs were divided into a control group, a TNF-α model group (referred to as model group) and TNF-α + PEMF group (referred to as PEMF group) and treated accordingly. Eight weeks after the modeling, safranin-o/fast green staining was used to assess any pathological morphology changes. The expression of NPY, neuropeptide Y receptor Y2 (NPY2R), bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), collagen type II (Col-II) and matrix metalloproteinase-3 (MMP3) in the intervertebral disc and the cultivated nucleus pulposus cells of the 3 groups were determined. Results:The intervertebral disc cells in the model group were ruptured and folded, with significantly increased polysaccharide and protein components, and significantly increased bone fibers. In the PEMF group the cell boundaries were clearer, with less fibrin fracture and increased cartilage tissue. NPY was expressed in the fibrous annulus and the nucleus pulposus of the intervertebral disc in the model group. The average expression levels of NPY and NPY2R were significantly higher than in the control group and the model group. Compared with the control group, there was a significant increase in the level of Bax and a significant decrease in the expression of Bcl-2 in the model group, and there was a significant decrease in the level of Bax in the PEMF group. Compared with the control group, there was a significant decrease in the Col-II level but a significant increase in the MMP3 protein expression in the model group. The average Col-II mRNA expression was significantly higher in the PEMF group compared with the model group, but the average MMP3 protein expression was significantly less. Those results are consistent with observations in vivo.Conclusion:PEMF may reverse the imbalance of ECM metabolism and delay IDD degeneration by up-regulating the expression of NPY and Bcl-2, as well as blocking the Bax/Bcl-2 signaling pathway to inhibit apoptosis of nucleus pulposus cells.

Objective:To explore any effect of pulsed electromagnetic field (PEMF) stimulation on intervertebral disc degeneration (IDD).Methods:Forty Sprague-Dawley rats were randomly divided into a control group, an IDD model group, a PEMF group and an observation group. An IDD model was induced in all except those in the control group. Both the PEMF and observation groups were given PEMF stimulation, while the latter was additionally injected with the A2AR agonist CGS-21680. Eight weeks after the modelling any pathological changes in the morphology of the rats′ intervertebral disc tissues were evaluated using saffron solid green staining. The expression of A2AR, cyclic adenosine phosphate (cAMP), protein kinase A (PKA), cysteine aspartate proteolytic enzyme-3 (Caspase-3), type II collagen (COL-II) and matrix metalloproteinase-3 (MMP3) in the intervertebral discs were evaluated.Results:The nucleus pulposus had shrunk, while fibrous tissues and chondrocytes had increased in the IDD model group. In the observation group the nucleus pulposus was intact and of basically normal shape. A2AR mRNA and protein levels were higher in the intervertebral disc tissue of the model group than among the control group, on average, while the levels in the observation group were significantly higher than in the other groups. In the PEMF and observation groups cAMP and PKA mRNA were significantly higher than in the IDD model group. The p38 MAPK and P-P38 MAPK levels of the IDD model group and its average P-P38 MAPK/p38 MAPK ratio were significantly higher than in the control group. In the PEMF and observation groups those indices had decreased to varying degrees, with those of the observation group significantly lower than among the model and PEMF groups on average, except for the p38 MAPK values. Caspase-3 and its mRNA were significantly higher in the model group than in the control group, on average, and those values were significantly lower in the PEMF and observation groups than in the IDD model group. The average MMP3 contents of the IDD model group had increased significantly compared with the control group, while the Col-Ⅱ level had decreased significantly. Compared with the IDD model group, the MMP3 level had decreased but Col-Ⅱ expression had increased in both the PEMF and observation groups, with significant differences between the IDD model and observation groups.Conclusions:The activation of the p38 MAPK signaling pathway by inflammatory factors to induce apoptosis is one of the important reasons for the aggravation of IDD lesions. PEMF combined with A2AR agonists can activate the A2AR/cAMP/PKA signaling pathway, inhibit p38 MAPK phosphorylation, reduce apoptosis of nucleus pulposus cells, and relieve IDD damage.

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.

Deep integration of healthcare and prevention in public hospitals is not only a basic function played by hospitals in their public health services, but also an inevitable choice to meet the health needs of the people in their life span. The authors analyzed the current situation in healthcare and prevention integration in Wuhan in recent years, focusing on such problems existing in the construction of healthcare and prevention integration in public hospitals, as unclear functional positioning of medical prevention integration in public hospitals, insufficient refinement of healthcare and prevention integration policies, delay in the construction of public health informatization, and poor public health awareness of medical personnel. In view of the above problems, the authors put forward the following improvement suggestions: optimizing the policy environment of healthcare and prevention integration, strengthening the leading role of the hospital management, building a hospital public health big data platform, mobilizing the initiative of clinical technicians, and improving the work identity of hospital public health workers.

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.

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.

Objective: To explore the expression of the A_2A 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 A_2A 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 A_2A 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 A_2A adenosine receptor increased significantly, while the expression of IL-1β and TNF-α decreased significantly. However, the A_2A 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 A_2A adenosine receptor in those cells.