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As a member of transforming growth factor β (TGF-β) family, bone morphogenetic proteins (BMPs) are multi-functional factors and play critical roles in heart, cartilage, neural development and postnatal bone formation. It has been demonstrated that among the family, BMP2/4 have been reported to be especially important for the developmental and maturation of central nervous system (CNS). It has different, even opposite functions, in certain given circumstances, which could be a potential risk for BMPs' clinical use.骨形态发生蛋白(bone morphogenetic proteins，BMPs)属于转化生长因子-β(transforming growth factor β，TGF-β)超家族，在心、神经、软骨的发育以及出生后骨的形成等发育过程中均起重要的调节作用。近年来的研究表明：在各类BMPs分子中，BMP2/4参与并促进了发育期和成熟期的中枢神经系统内神经元与胶质细胞的发育与激活，且在特定的环境中发挥不同甚至相反的生物学作用，这种差异性为其临床应用带来了一定的风险。.

Septic myocardial dysfunction remains prevalent and raises mortality rate in patients with sepsis. During sepsis, tissues undergo tremendous oxidative stress which contributes critically to organ dysfunction. Edaravone, a potent radical scavenger, has been proved beneficial in ischemic injuries involving hypoxia-inducible factor- (HIF-) 1, a key regulator of a prominent antioxidative protein heme oxygenase- (HO-) 1. However, its effect in septic myocardial dysfunction remains unclarified. We hypothesized that edaravone may prevent septic myocardial dysfunction by inducing the HIF-1/HO-1 pathway. Rats were subjected to cecal ligation and puncture (CLP) with or without edaravone infusion at three doses (50, 100, or 200 mg/kg, resp.) before CLP and intraperitoneal injection of the HIF-1α antagonist, ME (15 mg/kg), after CLP. After CLP, rats had cardiac dysfunction, which was associated with deformed myocardium, augmented lipid peroxidation, and increased myocardial apoptosis and inflammation, along with decreased activities of catalase, HIF-1α, and HO-1 in the myocardium. Edaravone pretreatment dose-dependently reversed the changes, of which high dose most effectively improved cardiac function and survival rate of septic rats. However, inhibition of HIF-1α by ME demolished the beneficial effects of edaravone at high dose, reducing the survival rate of the septic rats without treatments. Taken together, edaravone, by inducing the HIF-1α/HO-1 pathway, suppressed oxidative stress and protected the heart against septic myocardial injury and dysfunction.

Background Endometrial cancers (ECs) are one of the most common types of malignant tumor in females. Substantial efforts had been made to identify significantly mutated genes (SMGs) in ECs and use them as biomarkers for the classification of histological subtypes and the prediction of clinical outcomes. However, the impact of non-significantly mutated genes (non-SMGs), which may also play important roles in the prognosis of EC patients, has not been extensively studied. Therefore, it is essential for the discovery of biomarkers in ECs to further investigate the non-SMGs that were highly associated with clinical outcomes. Results For the 9681 non-SMGs reported by the mutation annotation pipeline, there were 1053, 1273 and 395 non-SMGs differentially expressed between the patient groups divided by the clinical endpoints of histological grade, histological type as well as the International Federation of Gynecology and Obstetrics (FIGO) stage of ECs, respectively. In the gene set enrichment analysis, the cancer-related pathways, namely neuroactive ligand-receptor interaction signaling pathway, cAMP signaling pathway and calcium signaling pathway, were significantly enriched with the differentially expressed non-SMGs for all the three endpoints. We further identified 23, 19 and 24 non-SMGs, which were highly associated with histological grade, histological type and FIGO stage, respectively, from the differentially expressed non-SMGs by using the variable combination population analysis (VCPA) approach and found that 69.6% (16/23), 78.9% (15/19) and 66.7% (16/24) of the identified non-SMGs had been previously reported to be correlated with cancers. In addition, the averaged areas under the receiver operating characteristic curve (AUCs) achieved by the predictive models with identified non-SMGs as predictors in predicting histological type, histological grade, and FIGO stage were 0.993, 0.961 and 0.832, respectively, which were superior to those achieved by the models with SMGs as features (averaged AUCs = 0.928, 0.864 and 0.535, resp.). Conclusions Besides the SMGs, the non-SMGs reported in the mutation annotation analysis may also involve the crucial genes that were highly associated with clinical outcomes. Combining the mutation status with the gene expression profiles can efficiently identify the cancer-related non-SMGs as predictors for cancer prognostic prediction and provide more supplemental candidates for the discovery of biomarkers. Electronic supplementary material The online version of this article (10.1186/s12859-017-1891-6) contains supplementary material, which is available to authorized users.

Background. Glucose fluctuation confers additional risks on diabetes-related vascular diseases, but the underlying mechanisms are unknown. Macrophage activation mediated by TLR4-JNK signaling plays an important role during the progress of diabetes. In the present study, we hypothesize that glucose fluctuation results in macrophage inflammation through TLR4-JNK signaling pathways. Methods. THP-1 cells were treated with normal glucose (5 mM), constant high glucose (25 mM), and intermittent high glucose (rotation per 6 h in 5 mM or 25 mM) for 24 h. The mRNA and protein expression levels of TLR4, p-JNK, and adipocyte fatty acid-binding protein (A-FABP) were determined, and the proinflammatory cytokines TNF-α and IL-1β were quantified. Results. In constant high glucose, TLR4 expression and JNK phosphorylation levels increased, and this effect was more pronounced in intermittent high glucose. Accordingly, the expression of A-FABP and the release of the proinflammatory cytokines TNF-α and IL-1β also increased in response to constant high glucose, an effect that also was more evident in intermittent high glucose. The inhibition of p-JNK by SP600125 did not attenuate TLR4 expression, but totally inhibited both A-FABP expression and the production of the proinflammatory cytokines TNF-α and IL-1β in both constant and intermittent high glucose. Conclusions. Intermittent high glucose potentiates A-FABP activation and inflammatory responses via TLR4/p-JNK signaling in THP-1 cells. These findings suggest a more detrimental impact of glucose fluctuation on macrophage inflammation in diabetes-related vascular diseases than thus far generally assumed.

Aims: Although RNA-sequencing has been widely used to identify the differentially expressed genes (DEGs) as biomarkers to guide the therapeutic treatment, it is necessary to investigate the concordance of DEGs identified by microarray and RNA-sequencing for the clinical prognosis. Material & methods: By using The Cancer Genome Atlas data sets, we thoroughly investigated the concordance of DEGs identified from microarray and RNA-sequencing data and their molecular functions. Results: The DEGs identified by both technologies averaged ˜98.6% overlap. The cancer-related gene sets were significantly enriched with the DEGs and consistent between two technologies. Conclusions: The highly consistency of DEGs in their regulation directionality and molecular functions indicated the good reproducibility between microarray and RNA-sequencing in identifying potential oncogenes for clinical prognosis.

Rationale: Pain-related anxiety and depression are well known to be comorbid with chronic pain and adversely affect patient quality of life. Recent studies have shown that anxiety-like behaviors also develop with acute surgical pain, but the effects of general anesthetics on acute pain-related anxiety are unknown. Objective: The present study aimed to compare the effects of different general anesthetics on anxiety-like behaviors that follow formalin-induced acute pain in a rat model. Methods: Formalin-induced acute inflammatory pain was established by intraplantar injection of 1 % formalin without anesthesia or with anesthesia using the clinical anesthetics sevoflurane, propofol, or pentobarbital sodium. Anxiety-like behaviors were studied using the open-field test and elevated plus maze. Phosphorylated extracellular signal-regulated kinase (p-ERK) 1/2 expression in the anterior cingulate cortex (ACC) and spinal cord was examined using immunohistochemistry. Results: Anxiety-like behaviors were observed at 24 and 72 h post-formalin injection. Concomitantly, p-ERK 1/2 expression was upregulated in the ACC at 1 and 24 h post-formalin injection. While all three general anesthetics effectively blocked nociceptive responses and activation of ERK in the rat ACC following formalin injection during anesthesia, only sevoflurane inhibited ERK activation in the spinal cord and ACC at 24 h post-injection. Conclusions: This study suggests that sevoflurane, but not intravenous anesthetics, inhibits pain-related anxiety, along with ERK activation in the ACC, probably through inhibition of spinal nociceptive transmission. Intraoperative application of inhaled anesthetics may be a better choice to reduce postoperative anxiety. Refereed/Peer-reviewed

Endothelial dysfunction induced by oxidative stress and inflammation plays a critical role in the pathogenesis of cardiovascular diseases. The anesthetic sevoflurane confers cytoprotective effects through its anti-inflammatory properties in various pathologies such as systemic inflammatory response syndrome and ischemic-reperfusion injury but mechanism is unclear. We hypothesized that sevoflurane can protect against tumor necrosis factor (TNF)-α-induced endothelial dysfunction through promoting the production of endothelium-dependent nitric oxide (NO). Primary cultured human umbilical vein endothelial cells (HUVECs) were pretreated with different concentrations (0.5, 1.5 and 2.5 minimum alveolar concentration, MAC) of sevoflurane for 30 min before TNF-α (10 ng/mL) stimulation for 4 h. Sevoflurane pretreatment significantly reduced TNF-α-induced VCAM-1, ICAM-1, IκBα, and NF-κB activation, and blocked leukocytes adhesion to HUVECs. Meanwhile, sevoflurane (1.5 and 2.5 MAC) significantly induced endothelial nitric oxide synthase (eNOS) phosphorylation and enhanced NO levels both intracellularly and in the cell culture medium. All these cytoprotective effects of sevoflurane were abrogated by NG-nitro-l-arginine methyl ester (l-NAME), a non-specific nitric oxide synthase inhibitor. Collectively, these data indicate that sevoflurane protects against TNF-α -induced vascular endothelium dysfunction through activation of eNOS/NO pathway and inhibition of NF-κB.

Background Spinal anesthesia (SA) is usually associated with hypotension in pregnant women. We sought to assess the influence of various maternal positions on SA-induced hypotension. Material/Methods The study population comprised 99 women at full-term gestation scheduled for elective cesarean section. They were randomized into 3 equal groups: the LL group, in which the patient was placed in the full left-lateral position until the start of surgery with the Whitacre needle bevel oriented laterally; the LS group, in which the patient was placed in the full left-lateral position initially and then shifted to the left-tilt supine position with the needle bevel oriented laterally; and the CS group, in which the patient was initially placed in the full left-lateral position and then shifted to the left-tilt supine position with the needle oriented in the cephalad direction. Results The incidences of hypotension in the LL, LS, and CS groups were 9.7%, 54.8%, and 56.3%, respectively. Ephedrine requirements were lower in the LL group than in the LS group (P

To detect the effect of pulse radiofrequency (PRF) treatment on the neuropathic pain established by L5-spinal nerve ligation (SNL) on rats, and to investigate if PRF treatment would affect the expression of autophagy related protein LC3 and autophagy related receptor P62 at the dorsal horn. Methods: A total of 36 male Sprague-Dawley rats were randomly divided into 3 groups: a Sham group, a SNL group, and a SNL+PRF group. The 50% paw withdrawal mechanical threshold (PWMT) was detected at 1 day before and 1, 3, 7, 14 and 28 days post-operation by using Von-Frey filaments. The autophagy related protein LC3 and autophagy related receptor P62 were investigated by Western blot. Results: Compared with the Sham group, the PWMT significantly decreased in the SNL group at each time points (P0.05); in SNL+PRF group, PRF treatment could elevate the PWMT at the 1st day post-operation and lasted for 28 days (P0.05). What's more, SNL could elevate the LC3-II and P62 levels at the 7th day post-operation (P0.05), which were decreased by the PRF treatment (P0.05). Conclusion: PRF treatment could improve SNL-induced the neuropathic pain, which might be partly due to the regulatory effects on the autophagy levels at the spinal dorsal horn.目的：观察腰5(L5)脊神经结扎(spinal nerve ligation，SNL)致慢性疼痛大鼠接受背根神经节脉冲射频(pulse radiofrequency，PRF)治疗后痛觉行为学的变化，检测脊髓背角自噬相关基因LC3和自噬受体蛋白P62的表达情况，研究PRF是否能通过调节脊髓背角神经元自噬水平发挥镇痛作用。方法：36只SD大鼠随机分为假手术组(Sham组)、SNL组(行左侧L5脊神经的暴露和结扎)和SNL+PRF组(建立SNL模型后即刻给予1次同侧背根神经节PRF治疗)。分别于手术前1 d及术后1，3，7，14，28 d观察各组大鼠机械刺激缩足反应阈值(paw withdrawal mechanical threshold，PWMT)改变，Western印迹检测自噬相关基因LC3和细胞自噬受体蛋白P62在大鼠损伤侧相应节段脊髓背角的表达改变。结果：与Sham组相比，SNL组大鼠在术后各时点PWMT均明显下降(P0.05)；与SNL组相比，SNL+PRF组大鼠在PRF治疗术后第1天即出现PWMT的升高，并持续28 d(P0.05)；与Sham组相比，SNL组大鼠在术后7 d脊髓背角LC3-II和P62蛋白表达明显升高(P0.05)；与SNL组相比，SNL+PRF组大鼠在术后7 d脊髓背角LC3-II和P62蛋白表达明显下调(P0.05)。 结论：SNL慢性疼痛大鼠接受背根神经节PRF治疗后出现疼痛行为学改善，PRF治疗可能通过上调脊髓背角神经元自噬水平发挥镇痛作用。.

MicroRNA (miRNA) plays an important role in the degradation and inhibition of mRNAs and is a kind of essential drug targets for cancer therapy. To facilitate the clinical cancer research, we proposed a network-based strategy to identify the cancer-related miRNAs and to predict their targeted genes based on the gene expression profiles. The strategy was validated by using the data sets of acute myeloid leukemia (AML), breast invasive carcinoma (BRCA), and kidney renal clear cell carcinoma (KIRC). The results showed that in the top 20 miRNAs ranked by their degrees, 90.0% (18/20), 70.0% (14/20), and 70.0% (14/20) miRNAs were found to be associated with the cancers for AML, BRCA, and KIRC, respectively. The KEGG pathways and GO terms enriched with the genes that were predicted as the targets of the cancer-related miRNAs were significantly associated with the biological processes of cancers. In addition, several genes, which were predicted to be regulated by more than three miRNAs, were identified to be the potential drug targets annotated by using the human protein atlas database. Our results demonstrated that the proposed strategy can be helpful for predicting the miRNA-mRNA interactions in tumorigenesis and identifying the cancer-related miRNAs as the potential drug targets.

To establish the improved methods for oligodendrocyte precursor cells (OPCs) isolation, culture and subculture and to investigate the characteristics of OPCs differentiation and proliferation in vitro. Methods: OPCs isolated from neonatal SD rat spinal cords were cultured in vitro. Cell proliferation was induced by the culture solution with basic fibroblast growth factor and platelet-derived growth factor, and the proliferation of OPCs was determined by MTT assay. OPCs differentiation to mature oligodendrocytes was induced by triiodothyronine and ciliary neurotrophic factor. The morphology and characteristics of OPCs and differentiated cells were observed via microscope and immunostaining. Results: More than 95% of the cells exhibited typical OPCs morphology with bipolar or tripolar bumps and expressed A2B5, while the OPCs kept the proliferative capacity in vitro. After culture for differentiation, the OPCs progressively differentiated into mature oligodendrocytes which specifically expressed myelin basic protein. Conclusion: The OPCs retain the ability of differentiation and proliferation in vitro.目的：对少突胶质前体细胞(oligodendrocyte precursor cells，OPCs)的分离、培养及传代方法进行改良，探讨体外条件下OPCs的增殖与分化特性。方法： 取新生SD大鼠脊髓来源的细胞分离、纯化OPCs行原代培养，用含碱性成纤维细胞生长因子(basic fibroblast growth factor，bFGF)和血小板源性生长因子-aa(platelet-derived growth factor-aa，PDGF-aa)的培养液作扩增培养，MTT法检测OPCs的增殖能力；三碘甲腺原氨酸和睫状神经生长因子诱导OPCs分化，最终分化为成熟的少突胶质细胞(oligodendrocytes，OLs)，并对分化细胞行形态学及免疫化学染色法鉴定。结果：95%以上的细胞具有双极或三极突起的典型OPCs形态并表达A2B5；MTT法检测显示OPCs在体外保持良好的增殖能力；细胞最终诱导分化为成熟的OLs并表达髓鞘碱性蛋白(myelin basic protein，MBP)。结论：OPCs在体外条件下可继续保持良好的增殖生长及定向分化能力。.

Sepsis-induced myocardial dysfunction increases mortality in sepsis, yet the underlying mechanism is unclear. Brain-derived neurotrophic factor (BDNF) has been found to enhance cardiomyocyte function, but whether BDNF has a beneficial effect against septic myocardial dysfunction is unknown. Septic shock was induced by cecal ligation and puncture (CLP). BDNF was expressed in primary cardiomyocytes, and its expression was significantly reduced after sepsis. In rats with sepsis, a sharp decline in survival was observed after CLP, with significantly reduced cardiac BDNF expression, enhanced myocardial fibrosis, elevated oxidative stress, increased myocardial apoptosis, and decreased endothelial nitric oxide (NO) synthase (eNOS) and NO. Supplementation with recombined BDNF protein (rhBDNF) enhanced myocardial BDNF and increased survival rate with improved cardiac function, reduced oxidative stress, and myocardial apoptosis, which were associated with increased eNOS expression, NO production, and Trk-B, a BDNF receptor. Pretreatment with NOS inhibitor, N (omega)-nitro-L-arginine methyl ester, abolished the abovementioned BDNF cardioprotective effects without affecting BDNF and Trk-B. It is concluded that BDNF protects the heart against septic cardiac dysfunction by reducing oxidative stress and apoptosis via Trk-B, and it does so through activation of eNOS/NO pathway. These findings provide a new treatment strategy for sepsis-induced myocardial dysfunction.

Sepsis-induced cardiac dysfunction remains one of the major causes of death in intensive care units. Overwhelmed inflammatory response and unrestrained cell death play critical roles in sepsis-induced cardiac dysfunction. Peroxisome proliferator-activated receptor- (PPAR-) γ has been proven to be cardioprotective in sepsis. However, the mechanism of PPAR-γ-mediated cardioprotection and its relationship with inflammation and cell death are unclear. We hypothesized that activation of PPAR-γ by reducing cardiac inflammation, myocardial apoptosis, and necroptosis may prevent myocardial dysfunction in sepsis. Rats were subjected to cecal ligation and puncture (CLP) with or without PPAR-γ agonist (rosiglitazone) or antagonist T0070907 (T007). After CLP, cardiac function was significantly depressed, which was associated with the destructed myocardium, upregulated proinflammatory cytokines, and increased apoptosis, necrosis, and necroptosis. This process is corresponded with decreased inhibitor κB (IκBα) and increased NF-κB, receptor-interacting protein kinase-1 (RIP1), RIP3, and mixed lineage kinase-like (MLKL) protein. Activation of PPAR-γ by rosiglitazone pretreatment enhanced PPAR-γ activity and prevented these changes, thereby improving the survival of septic rats. In contrast, inhibition of PPAR-γ by T007 further exacerbated the condition, dropping the survival rate to nearly 0%. In conclusion, PPAR-γ activation by reducing proinflammatory cytokines, apoptosis, and necroptosis in the myocardium prevents septic myocardial dysfunction.

Opioid addiction is a major social, economic, and medical problem worldwide. Long-term adverse consequences of chronic opiate exposure not only involve the individuals themselves but also their offspring. Adolescent maternal morphine exposure results in behavior and morphologic changes in the brain of their adult offspring. However, few studies investigate the effect of adult opiate exposure on their offspring. Furthermore, the underlying molecular signals regulating the intergenerational effects of morphine exposure are still elusive. We report here that morphine exposure of adult male and female rats resulted in anxiety-like behavior and dendritic retraction in the dentate gyrus (DG) region of the hippocampus in their adult offspring. The behavior and morphologic changes were concomitant with the downregulation of insulin-like growth factor (IGF)-2 signaling in the granular zone of DG. Overexpression of hippocampal IGF-2 by bilateral intra-DG injection of lentivirus encoding the IGF-2 gene prevented anxiety-like behaviors in the offspring. Furthermore, exposure to an enriched environment during adolescence corrected the reduction of hippocampal IGF-2 expression, normalized anxiety-like behavior and reversed dendritic retraction in the adult offspring. Thus, parental morphine exposure can lead to the downregulation of hippocampal IGF-2, which contributed to the anxiety and hippocampal dendritic retraction in their offspring. An adolescent-enriched environment experience prevented the behavior and morphologic changes in their offspring through hippocampal IGF-2 signaling. IGF-2 and an enriched environment may be a potential intervention to prevention of anxiety and brain atrophy in the offspring of parental opioid exposure. Refereed/Peer-reviewed

Lidocaine, as an anesthetic substance, is often used for surface and spinal anesthesia. However, studies have shown that lidocaine may induce transient neurological symptoms and cauda equina syndrome. In the present study the effects of the ginsenoside Rg1 (Rg1) on lidocaine‑induced apoptosis were assessed in Jurkat cells using flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The data showed that incubation with Rg1 provides protection against lidocaine‑induced apoptosis in cultured Jurkat cells. In order to investigate the effect of Rg1 on the apoptosis pathway, caspase 3 gene expression was determined. The results suggested that the protective effect of Rg1 on lidocaine‑induced apoptosis is mediated by altering the level of B‑cell lymphoma‑2 (BCL‑2) family proteins and downregulating caspase‑3 expression. In conclusion, the present study demonstrated that incubation with Rg1 provides protection against lidocaine‑induced apoptosis in cultured Jurkat cells. In addition, the study demonstrated that Rg1 is a notable antiapoptotic molecule that is capable of blocking the caspase‑dependent signaling cascade in Jurkat cells, and that the protective effect of Rg1 on lidocaine‑induced apoptosis is mediated by altering levels of BCL‑2 family proteins and downregulating caspase‑3 expression. The present study provides the basis for understanding and evaluating the effect of Rg1 in the in vivo treatment of lidocaine-induced transient neurological symptoms and cauda equina syndrome by lidocaine.

Reperfusion injury is tissue damage caused by the re-supply of blood following a period of ischemia in tissues. Intestinal ischemia-reperfusion injury (IRI) is an extremely common clinical event associated with distant organ injury. The intestine serves as the initial organ of multi-system organ dysfunction syndrome. It is extremely important to identify a method to protect against IRI, as it is a key factor associated with morbidity and mortality in patients. In the present study, the protective effects of pretreatment with dexmedetomidine hydrochloride were investigated. Rats were divided into six groups and models of intestinal ischemia were created in the five groups. Certain groups were pretreated with dexmedetomidine hydrochloride. The levels of TNF-α and IL-6 were measured by enzyme-linked immunosorbent assay in order to evaluate the injury. Tissue sections were stained with hematoxylin and eosin to visualize the damage. qPCR and western blotting were performed to examine the inflammatory status. Pretreatment with various doses of dexmedetomidine hydrochloride significantly reduced the pathological scores and the inflammatory reaction. The levels of TNF-α, IL-6, TLR4 and MyD88 were decreased in the dexmedetomidine hydrochloride treatment groups compared with those in the sham control and untreated ischemia reperfusion groups. The results of the present study indicate that pretreatment with dexmedetomidine hydrochloride may be a useful method of reducing the damage caused by IRI.

To investigate the effect of limb remote ischemic preconditioning (RIPC) on hepatic ischemia/reperfusion (IR) injury and the underlying mechanisms. Rats were subjected to partial hepatic IR (60 min ischemia followed by 24 hours reperfusion) with or without RIPC, which was achieved by 3 cycles of 10 min-occlusion and 10 min- reperfusion at the bilateral femoral arteries interval 30 min before ischemia. Some rats were treated with a new PPAR-γ inhibitor, T0070907, before RIPC. At the end of reperfusion, liver injury was significantly increased (increases in Suzike's injury score, AST and ALT release), concomitant with elevated oxidative stress (increases in MDA formation, MPO activity, as well as the decrease in SOD activity) and inflammation (increases in TNF-α and IL-6 levels, decrease in IL-10 content). RIPC improved liver function and reduced histologic damage, accompanied by the increased PPAR-γ activation and autophagosome formation as well as the reduced autophagosome clearance. The beneficial effects of RIPC were markedly attenuated by T0070907, an inhibitor of PPAR-γ. RIPC exerts the protective effects on liver by activation of autophagy via PPAR-γ.目的：探讨肢体远程缺血预处理(remote ischemic preconditioning，RIPC)对肝脏缺血/再灌注损伤的影响及其机制。方法：对大鼠进行局部的肝组织缺血再灌注(缺血60 min后再灌注24 h)，缺血前30 min通过3个循环的双侧股动脉闭塞10 min和再灌注10 min来实现RIPC。在RIPC之前，用PPAR-γ抑制剂T0070907处理部分大鼠。结果：在再灌注结束时，局部缺血后肝损伤显著增加Suzike的损伤评分和AST及ALT释放，并伴随有氧化应激和炎症反应增加。RIPC能够改善肝脏缺血/再灌注后的肝功能，减少肝组织病理损伤，与PPAR-γ活化和自噬体增多相关。PPAR-γ抑制剂T0070907显著减少自噬体形成，抑制RIPC对肝脏缺血/再灌注损伤的保护作用。结论：肢体远程缺血预处理是通过活化的PPAR-γ激活肝脏自噬，从而对肝脏起保护作用。.

The interactions among the genes within a disease are helpful for better understanding the hierarchical structure of the complex biological system of it. Most of the current methodologies need the information of known interactions between genes or proteins to create the network connections. However, these methods meet the limitations in clinical cancer researches because different cancers not only share the common interactions among the genes but also own their specific interactions distinguished from each other. Moreover, it is still difficult to decide the boundaries of the sub-networks. Therefore, we proposed a strategy to construct a gene network by using the sparse inverse covariance matrix of gene expression data and divide it into a series of functional modules by an adaptive partition algorithm. The strategy was validated by using the microarray data of three cancers and the RNA-sequencing data of glioblastoma. The different modules in the network exhibited specific functions in cancers progression. Moreover, based on the gene expression profiles in the modules, the risk of death was well predicted in the clustering analysis and the binary classification, indicating that our strategy can be benefit for investigating the cancer mechanisms and promoting the clinical applications of network-based methodologies in cancer researches.

The precursor of brain derived neurotrophic factor (proBDNF), the unprocessed BDNF gene product, binds to its receptors and exerts the opposing biologic functions of mature BDNF. proBDNF is expressed in the peripheral tissues but the functions of peripheral proBDNF remain elusive. Here we showed that proBDNF and its predominant receptor, p75 pan-neurotrophin receptor were upregulated in the nerve fibers and inflammatory cells in the local tissue in inflammatory pain. Neutralization of proBDNF by polyclonal antibody attenuated pain in different models of inflammatory pain. Unilateral intra-plantar supplementation of proBDNF by injecting exogenous proBDNF or ectopic overexpression resulted in pain hypersensitivity and induced spinal phosphorylated extracellular signal-regulated kinase activation. Exogenous proBDNF injection induced the infiltration of inflammatory cells and the activation of proinflammatory cytokines, suggesting that inflammatory reaction contributed to the pro-algesic effect of proBDNF. Finally, we generated monoclonal anti-proBDNF antibody that could biologically block proBDNF. Administration of monoclonal Ab-proBDNF attenuated various types of inflammatory pain and surgical pain. Thus, peripheral proBDNF is a potential pain mediator and anti-proBDNF pretreatment may alleviate the development of inflammatory pain.

Hypothermia reduces immediate paralysis during surgical repair of aortic aneurysms. However, it is unknown what the impact of hypothermia is on delayed paralysis, a serious complication of this type of surgery.Sprague-Dawley rats were subjected to occlusion of the descending aorta at different duration under normothermia (38.0±0.5) or hypothermia (33.0±0.5°). Neurologic function was assessed. Motor neuron number, glial activation, and cytokine expression in the spinal cord were examined. Minocycline was administered perioperatively by intraperitoneal injection in the rats subjected to the aorta occlusion.In contrast to normothermia conditions at which immediate paralysis occurred when the duration of aorta occlusion exceeded 11.5min, hypothermia did not induce immediate paralysis if the duration of aorta occlusion was less than 41min. However, delayed paralysis was developed when the duration of aorta occlusion exceeded 18min, and reached peak level when the duration of aorta occlusion was 40min at hypothermia condition. The number of motoneurons was significantly decreased (P0.05) at 30h postoperation. In addition, microglia was activated, and interleukin-1β and interleukin-6 levels were upregulated, both of which were co-localized in microglia at 24h postoperation in the hypothermia group. Minocycline treatment attenuated the incidence and degree of paralysis but did not decrease the mortality.Hypothermia, a neuroprotective strategy in cardiothoracic surgery, increased the incidence of delayed paralysis through activation of spinal microglia and cytokines. Blocking the activated microglia may be a potential intervention to prevent the incidence of delayed paralysis.

Background The directional flow of injection through a Whitacre needle can be used to modify the level of sensory blockade. We hypothesized that injection of hyperbaric local anesthetic through a Whitacre needle with the bevel oriented laterally can produce a more symmetric sensory block. Methods Patients scheduled for lower limb surgery under spinal anesthesia with the patient in lateral decubitus position were randomized to receive 10 mg, 0.5% hyperbaric bupivacaine with the Whitacre needle orifice in 1 of 2 orientations, cephalad and lateral. The patient's position was maintained for 15 min after the injection, and sensory blocks were recorded. The primary outcome was the sensory levels between the dependent and nondependent side. The Wilcoxon-Mann-Whitney U-test odds was used to compare unpaired nonparametric data. For the paired samples, 95% confidence intervals (CI) of differences between group medians were calculated using the Hodges-Lehman estimator for the median difference in number of blocked segments. Results There was no significant difference in block level between dependent and nondependent sides in the lateral group. The Hodges-Lehmann point estimator was 0.5% and 95% CI was 0-2.5, suggesting a more symmetric sensory block in patients in the lateral decubitus position maintained for a sufficient period of time. A significantly lower level of blockade was noticed on the nondependent side compared to the dependent side in the cephalad group. The Hodges-Lehmann point estimator was 2.5 segments and 95% CI was 0.5-5. Conclusions Injection of 10 mg of 0.5% hyperbaric bupivacaine with the bevel of the Whitacre needle oriented laterally produces more symmetric sensory levels of blockade between the dependent and nondependent sides.

Objectives. The incidence of fentanyl-induced cough (FIC) during induction of general anesthesia varies around 40% and is undesirable. It increases intracranial, intraocular, and intra-abdominal pressures. This prospective, randomized, double-blind, placebo-controlled study evaluated the effect of dexmedetomidine (DEX) pretreatment on the incidence and severity of FIC. Methods. Altogether 300 patients undergoing elective surgical procedures were randomly allocated into three groups (I, II, III; n = 100) and administered intravenously, over 10 min, 10 mL isotonic saline, DEX 0.5 μg/kg in 10 mL isotonic saline, or DEX 1 μg/kg in 10 mL isotonic saline, respectively. All groups subsequently received a fentanyl (4.0 μg/kg) intravenous push. The incidence and severity of cough were recorded for 1 min after fentanyl administration. Results. The incidence of FIC was 61%, 40%, and 18% in groups I, II, and III, respectively (P < 0.05 for treatment groups II and III versus control group I). There was no significant difference in the severity or onset time of cough, or hemodynamic variables, among the three groups. Conclusions. Intravenous DEX (0.5 μg/kg or 1 μg/kg) immediately before the administration of intravenous fentanyl (4.0 μg/kg) significantly reduced the incidence of FIC.

Background Animal studies have shown that surgical trauma activates the descending noradrenergic pathway. However, perioperative patients have decreased concentrations of noradrenaline (NA) in the cerebrospinal fluid (CSF). We proposed that the descending monoaminergic pathway is altered in post-traumatic pain patients and that CSF monoamine neurotransmitters may be more closely related to affective symptoms. We investigated the levels of monoamine neurotransmitters and assessed pain in these patients. Methods Patients were divided into a post-traumatic pain group, a pain-free group, a painful labor group, and a pain-free labor group. CSF was collected from all patients, and concentrations of NA, 3-methoxy-4-hydroxyphenylglycol (MHPG), dopamine, homovanillic acid, and 5-hydroxyindoleacetic acid (5-HIAA) were measured by high-performance liquid chromatography. Results In the post-traumatic pain group, lumbar CSF concentrations of NA and MHPG were significantly decreased (P < 0.01) compared to the control group. The post-traumatic pain group displayed a significant negative correlation between NA and the respective total value of the short form of the McGill pain questionnaire (SF-MPQ), SF-MPQ (affective), and visual analog scale (r = –0.388, r = –0.433, and r = –0.367; P < 0.05). Conclusions Post-traumatic pain patients demonstrated decreased concentrations of NAin CSF, indicating that descending noradrenergic pain control pathways may be inhibited. NA is more closely related to negative affects in post-traumatic pain patients.

Aims Endothelial cell injury induced by inflammatory factors plays a critical role in the pathogenesis of numerous vascular diseases. MicroRNAs are well known to be implicated in cell proliferation and apoptosis in inflammatory responses; however, it remains to be determined whether microRNAs are associated with tumour necrosis factor (TNF)-α-mediated endothelial cell injury. The aim of the present study was to investigate the role of microRNAs in TNF-α-induced endothelial cell apoptosis. Methods and results Microarrays were used to analyse the global expression of microRNAs in TNF-α-stimulated human primary endothelial cells. Expression profiles of the microRNAs were verified using qRT-PCR. After TNF-α treatment, 12 miRNAs were dramatically up-regulated and nine were down-regulated. LNA-anti-miR-23a and pre-miR-23a were found to modulate one of the markedly down-regulated miRNAs, miR-23a, which could in turn increase or attenuate TNF-α-induced endothelial cell apoptosis. Bioinformatics analysis suggested that caspase-7 and serine/threonine kinase 4 are potential targets of miR-23a. LNA-anti-miR-23a enhanced but pre-miR-23a inhibited the activation of caspase-7, serine/threonine kinase 4, and its related signalling caspase-3 after TNF-α treatment; however, neither pre-miR-23a nor LNA-anti-miR-23a had an effect on TNF-α-induced Bcl-2 activation. Conclusion Our results suggest that miR-23a may be involved in TNF-α-induced endothelial cell apoptosis through regulation of the caspase-7 and serine/threonine kinase 4–caspase-3 pathways.

The objective of this study was to investigate, in a rabbit model, the cardioprotective effect of morphine preconditioning against cardiac remodelling.In male New Zealand rabbits myocardial I/R injury was established by ligating the left descending coronary artery followed by perfusion. The rabbits were randomly divided into: (i) a normal saline sham group (S), (ii) a morphine sham group (MS), (iii) a normal saline group (NS); (iv) a morphine preconditioning group (MPC). After I/R injury, cardiac function was evaluated by echocardiography and biologic markers at day 1, day 3, day 7, day 14, day 21 and day 28. At day 1 after I/R injury, in comparison to the NS group, morphine preconditioning: (i) significantly improved haemodynamic parameters; (ii) reduced myocardial infarct size; (iii) lowered troponin I level. At day 28 after I/R injury, the concentration of B-type natriuretic peptide was significantly lower in the MPC group compared to the NS group. The MPC group had heavier left ventricles than the MS group while the NS group had heavier right ventricles than the other groups, respectively. Furthermore, the MPC group showed improved haemodynamic parameters. Echocardiography showed a higher left ventricular ejection fraction and fractional shorting in the MPC group compared to the NS group at each observation point.After I/R injury and morphine preconditioning by a single dose, we have observed a prolonged cardioprotective effect and an improved cardiac function. Morphine preconditioning may reduce post-infarction remodelling.

Background: It is still not known whether the spinal cytokine signaling pathways are involved in the pathophysiologic mechanism of the acute phase of heart disease. This study examines the expression pattern of tumor necrosis factor-α (TNF-α) and its two related mitogenic-activated protein kinases, p38 and Jun-N-terminal kinase (JNK), in the spinal cord in response to acute cardiac injury (ACI). Methods: The ACI rat model was established by intra-myocardial injection of formalin. At the indicated times after the establishment of ACI, the thoracic segments of the spinal cord were harvested and Western blot was performed to determine the expression of TNF-α, p38 and JNK. The localization of the cytokine and the kinases was determined by immunohistochemistry and double immunofluorescence. Results: In response to ACI, TNF-α protein was up-regulated and reached a peak level at 6 h after ACI. The up-regulated TNF-α was distributed in all the laminae in the spinal cord and mainly localized in the neurons, as determined by immunohistochemistry and double immunofluorescence. In response to ACI, p38 and JNK were also up-regulated in the spinal cord. The expression profiles of p38 and JNK were similar to that of activated TNF-α following ACI. Conclusions: This study shows that cardiac injury can induce the activation of spinal TNF-α, p38 and JNK. The activated spinal cytokine signaling may contribute to disease progression in the acute phase of cardiac injury in clinical practice.

Pulmonary fibrosis is a severe disease that contributes to the morbidity and mortality of a number of lung diseases. However, the molecular and cellular mechanisms leading to lung fibrosis are poorly understood. This study investigated the roles of epithelial-mesenchymal transition (EMT) and the associated molecular mechanisms in bleomycin-induced lung fibrosis. The bleomycin-induced fibrosis animal model was established by intratracheal injection of a single dose of bleomycin. Protein expression was measured by Western blot, immunohistochemistry, and immunofluorescence. Typical lesions of lung fibrosis were observed 1 week after bleomycin injection. A progressive increase in MMP-2, S100A4,α-SMA, HIF-1α, ZEB1, CD44, phospho-p44/42 (p-p44/42), and phospho-p38 MAPK (p-p38) protein levels as well as activation of EMT was observed in the lung tissues of bleomycin mice. Hypoxia increased HIF-1αand ZEB1 expression and activated EMT in H358 cells. Also, continuous incubation of cells under mild hypoxic conditions increased CD44, p-p44/42, and p-p38 protein levels in H358 cells, which correlated with the increase in S100A4 expression. In conclusion, bleomycin induces progressive lung fibrosis, which may be associated with activation of EMT. The fibrosis-induced hypoxia may further activate EMT in distal alveoli through a hypoxia-HIF-1α-ZEB1 pathway and promote the differentiation of lung epithelial cells into fibroblasts through phosphorylation of p38 MAPK and Erk1/2 proteins.

The surface properties and flotation behaviour of aluminosilicates including kaolinite, illite and pyrophyllite, using N-(3-aminopropyl)-dodecanamide as a collector, have been investigated. The best recoveries of kaolinite, illite and pyrophyllite were respectively 91.5%, 90.6%, and 96.3%. On the basis of FT-IR spectra and zeta potential, the ionization of surface hydroxyl, as well as isomorphic exchange of surface ions, accounts for the mineral surface charges. The mechanisms of interactions of the mineral surface with collectors depend on the pH of the pulp. In the acidic region, the interactions of the mineral surfaces with the collector were mainly electrostatic, but in the alkaline region, the interactions are mainly due to hydrogen bonding. It can be seen from SEM photographs of aluminosilicates that pyrophyllite takes the shape of thin slices, but kaolinite and illite have irregular porous structures. Specific surface area and particle size distributions of aluminosilicates also indicate that kaolinite and illite are relatively large porous particles with larger external as well as internal surface areas.

Background and objectives The overall purpose of this study was to investigate the role of cytochrome C oxidase (CcO) in preventing ischemia reperfusion-induced cardiac injury through gaseous signaling molecule pathways. Materials and methods We used CcO inhibitor, potassium cyanide (KCN) to mimic the pre-treatment of gaseous signaling molecules in a global ischemia/reperfusion (IR) injury model in rats. Intracellular reactive oxygen species (ROS) was determined by measuring mitochondrial H2O2 and mitochondrial complex activity. Results KCN pre-treatment led to decreased infarction area after IR injury and improved cardiac function. KCN pre-treated group challenged with IR injury was associated with reduced ROS production through inhibition of activity and not downregulation of CcO expression. In addition, KCN pre-treatment was associated with enhanced expression and activity of mitochondrial antioxidase, suggesting the role of CcO in regulating IR injury through oxidative stress. Conclusion KCN pre-treatment reduced the severity of IR injury. The potential mechanism could be increased endogenous anti-oxidase activity and consequently, the enhanced clearance of ROS.

Background Remifentanil, an ultra-short-acting opioid, is widely used for pain control during surgery. However, regular dose (RD) remifentanil exacerbates postoperative pain in a dose-dependent manner. Recent studies suggest that high-dose (HD) remifentanil offers sustained analgesia in experimental studies. We thus hypothesized that intraoperative administration of high-dose remifentanil may attenuate postoperative pain. Methods In this prospective, randomized, double blind, controlled clinical study, sixty patients undergoing thyroidectomy (18–60 years-of-age) received an intraoperative infusion of 0.2 (RD group) or 1.2 μg kg−1min−1 (HD group) remifentanil during thyroidectomy. A visual analogue scale (VAS) was used to measure pain intensity. Mechanical pain threshold on the forearm was assessed using von Frey filaments before surgery (baseline), 2 h postoperatively and 18–24 h postoperatively. The primary outcome was to compare the difference of VAS score at different time points after operation and morphine consumption 24 h postoperatively between RD and HD groups. The second outcome was to compare the difference of mechanical pain thresholds in the forearm postoperatively between RD and the HD groups. Results VAS scores were lower 30 min postoperatively in the HD group (1.29±1.67, 95% CI 0.64–1.94) compared with the RD group (2.21±1.67, 95% CI 1.57–2.84) (t = 3.427, p = 0.0043, RD group vs. HD group). Postoperative morphine consumption was much lower in the HD group compared with the RD group (1.27±1.88 mg vs. 0.35±1.25 mg, p = 0.033). In both groups, mechanical pain threshold was decreased 18–24 h postoperatively (2.93±0.209 Ln(g) vs. 3.454±2.072 Ln(g), p = 0.032 in RD group; 2.910±0.196 Ln(g) vs. 3.621±0.198 Ln(g), p = 0.006 in HD group, 18–24 h postoperatively vs baseline). Conclusions Intraoperative administration of high-dose remifentanil decreased VAS scores and morphine consumption postoperatively. Thus, modulation of intraoperative opiates may be a simple and effective method of postoperative pain management. Trial Registration This trial is registered in ClinicalTrials.gov, with the Name: Effect of Higher Doses of Remifentanil on Postoperative Pain in Patients Undergoing Thyroidectomy, and ID number: NCT01761149.

To construct dopamine D1 receptor (DRD1) expression interference vectors to study the role of DRD1 in nerve cells and lay a foundation for drug development in anti-convulsion.Based on DRD1 gene sequence in GenBank, 10 interfere vectors of DRD1 were designed. Liposomal was used to transfect NG-108-15 and the transfect effect was assayed by GFP. With realtime PCR and Western blot, the DRD1 expression was detected.The 10 constructed interfere vectors transfected into NG-108-15 cells by liposomal method and inhibited DRD1 mRNA and protein expression. DRD1 mRNA expression in NG-108-15 cells transfected with pGPU6-GFP-Neo-si-DRD1-5 was the lowest whereas DRD1 protein expression in NG-108-15 cells transfected with pGPU6-GFP-Neo-si-DRD1-1, -2, -6, -7 was the lowest.DRD1 expression interference vector is successfully constructed.

Right before the excystment of an Euplotes encysticus sawtooth-like folds appeared among the pellicle plasmalemma, the inner and outer alveolar membranes were still sticking together, and were not distinguishable. Microtubular layers already formed at the sites beneath the dorsal cortical pellicle corresponding to vegetative cells, but they still proceed to be organized on the ventral structures. Cristae, highly-tangled with tubular-type structures, appeared on the mitochondria, and were morphologically similar to that of vegetative cells. In the cortical ciliatures, such as ciliary shafts, kinetosomes, surrounding fibrillar cirral baskets, and attached structures of ciliatures, etc., they are different from those in resting cysts which are degenerated or lost. All the ciliature microtubules of ciliary shafts are of the 9+2 pattern, but the microtubule-like structure aggregates at triplet - microtubule centers of many kinetosmes, are still under various stages of differentiation. Microtubules beneath the kinetosomal rows are of a developmentally elongated stage; crowded chromatins of various shapes and sizes are found in macronucleus, but there are no nuclear pores (formed by nuclear membrane as in resting cysts) on the nuclear membrane where these chromatins attached.

The present work deals with the flotation behavior of kaolinite, illite and pyrophyllite with N-(2-aminoethyl)-1-naphthaleneacetamide (AENA). The best recoveries of kaolinite, illite and pyrophyllite were 41.4%, 39.8% and 90.4% respectively. The flotation behavior of three aluminosilicates is almost independent of pulp pH and the collector concentration. The results obtained show that AENA is a stronger collector for pyrophyllite. Conversely it is weaker for kaolinite and illite. By determining the effects of pulp pH on the zeta potential of aluminosilicates, we found that along with the rise in pulp pH, the zeta potential of the aluminosilicate pulp changed gradually from the plus zone to the minus zone. However, there is not a marked change in the floatability of the aluminosilicates. It was shown that the interaction between the collector and the surfaces of the ore particle was not merely electrostatic force, there are also probably hydrogen bonds between them.

To evaluate the role of transforming growth factor-β1 (TGF-β1) and connective tissue growth factor (CTGF) in the pathogenesis of pulmonary artery hypertension, we observed the dynamic expression of TGF-β1 and CTGF in rats with high blood flow.Fifty adult male SD rats were randomly divided into 5 groups: a sham group (group S) and groups with right pneumonectomy for 1, 2, 4 and 6 weeks (PE1, PE2, PE4 and PE6 group), 10 rats per group. The mean pulmonary arterial pressure (mPAP), vessel morphometry and right ventricle hypertrophy index (RVHI) were measured. TGF-β1 and CTGF protein expression in the lung tissues were determined with immunohistochemistry and Western blot. The expression of TGF-β1 mRNA and CTGF mRNA in the lung tissues was evaluated by RT-PCR.Compared with group S, the mPAP and RVHI in the rats were significantly increased in group PE1, PE2, PE4, and PE6 (P0.05); the indicators of vascular remodeling [(MA+PMA)%, RMT, and RMA] were markedly elevated in group PE4 and PE6 (P0.05), but not in group PE1 and PE2. Immunohistochemical staining of TGF-β1 and CTGF were more prominent in all of the right pneumonectomy groups than in the sham group. Western blot showed that the level of TGF-β1 protein was significantly increased in all of the right pneumonectomy groups (P0.01), and the peak was observed in group PE2, whereas the level of CTGF protein was markedly elevated in group PE4 and PE6 (P0.05), but no change was noticed in group PE1 and PE2. Compare to group S, the mRNA level of TGF-β1 was dramatically increased in all right pneumonectomy groups (P0.01), peaked at group PE2, and remained high in group PE4 and PE6. In contrast, the elevation of mRNA level of CTGF was not significant in group PE1, but group PE2, PE4 and PE6 demonstrated significant mRNA level of CTGF (P0.01). Correlation analysis showed that the protein and mRNA levels of CTGF were positively correlated with RMT and RMA ( r=0.743, r=0.906; P0.05), while no correlation between the protein and mRNA level of TGF-β1 with RMT or RMA. There was no correlation between the mRNA level of TGF-β1 and CTGF.TGF-β1 and CTGF play a role in the pathogenesis of increased pulmonary blood flow-induced pulmonary hypertension.

To elucidate the effects of mammalian sterile 20-like kinase 1 (MST1) gene on tumor necrosis factor (TNF)-α-mediated human umbilical vein endothelial cell (HUVEC) apoptosis.Cultured HUVECs were treated with either vehicle or TNF-α (1-100 ng/mL) for 24 hours. Cell apoptosis was measured by TUNEL staining, and MST1 activity was analyzed by Western blot. In order to knock down MST1 expression in HUVECs, cells were transfected with 100 nmol/L MST1 small interference RNA (siRNA) using Lipofectamine 2000 for 24 hours, and the transfection efficiency was analyzed by Western blot. MST1 siRNA-transfected cells were treated with 10 ng/mL TNF-α for an additional 24 hours. Cell apoptosis was measured by TUNEL staining and caspase-3 activity was detected by Western blot.MST1 activity was stimulated in a dose-dependent manner after TNF-α treatment (10, 40, 100 ng/mL) and reached the maximal effect at 100 ng/mL. MST1 activity also paralleled the onset of apoptosis as determined by TUNEL staining (P0.001). Transfection with MST1 siRNA markedly diminished MST1 gene expression in a dose-dependent manner. MST1 siRNA (100 nmol/L) significantly silenced MST1 gene (P0.05) and reduced TNF-α-induced endothelial cells apoptosis (P0.05) by way of inhibiting MST1 gene activation and, accordingly, suppressing caspase-3 activity.Silencing of MST1 expression by siRNA diminishes TNF-α-mediated human umbilical vein endothelial cell apoptosis by inhibiting the cascade effect of caspase-3.

This article is to investigate the role of outpatient informationization and the SAFE-CARE system in raising the level of outpatient high-quality medical service. It summarizes combination application experiences of outpatient informationization and the SAFE-CARE system to improve the level of outpatient medical service during the past year. Outpatient informationization has provided an efficient operational tool to improve the treatment efficiency; the SAFE-CARE system has guaranteed medical safety and quality, and enhanced the level of hospital management. Therefore, the combination application of outpatient informationization and the SAFE-CARE system is one of important measures to alleviate the difficulties of the masses medical treatment.

Quantitative analysis of biological image data generally involves the detection of many pixel spots. In live mitochondria video image, for which fluorescent microscopy is often used, the signal-to-noise ratio (SNR) can be extremely low, making the detection and tracking of mitochondria particle difficult. It is especially not easy to get the movement curve when the movement of the mitochondria involves its self-move and the motion caused by the neuron. An tracking algorithm for live mitochondria is proposed in this paper. First the whole image sequence is frame-to-frame registered, in which the edge corners are chosen to be the feature points. Then the mitochondria particles are tracked by frame-to-frame displacement vector. The algorithm proposed has been applied to the dynamic image sequence including neuron and mitochondria, saving time without manually picking up the feature points. It provides an new method and reference for medical image processing and biotechnological research.

Background Evidence suggests that increased leukotriene level and receptor protein expression emerged in the adenotonsillar tissue of children with sleep-disordered breathing (SDB). Nevertheless, few studies have investigated the contribution of disease severity in the cysteinyl leukotriene (CysLT) production and cysteinyl leukotriene receptor (CysLTR) expression. Furthermore, the relationship between local upper-airway and systemic inflammatory responses remains undefined. Methods A prospective, observational study that included standard questionnaires, physical examinations and overnight polysomnography. CysLTs were determined from serum, urine and tonsillar homogenate supernatant by means of enzyme-linked immunosorbent assays. The protein expressions of CysLTR were measured using Western blot analysis. Results Children with SDB had increased intratonsillar CysLT levels as well as CysLT subtype 1 receptor (CysLT1-R) and CysLT subtype 2 receptor (CysLT2-R) protein expression than the control group. CysLT concentration was positively correlated with body mass index z-score and apnea–hypopnea index (r = 0.454 and 0.487, p

In a previous study, we showed that a deep thoracic incision induces the segmental upregulation of interleukin-1β (IL-1β) in the spinal cord. However, whether the cytokines tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) are also activated in response to surgical incision remains to be determined. The present study aimed to investigate the expression pattern of TNF-α and IL-6 in the spinal cord following a deep thoracic incision. After surgical incision, the mRNA levels of TNF-α and IL-6 in the thoracic spinal cord were transiently upregulated as determined by real-time polymerase chain reaction (PCR) assay. However, the activation of IL-6 was detected at 1 h postoperatively, which was earlier compared to that of TNF-α, observed at 6 h postoperatively. The activated TNF-α was mainly localized in the neurons, but not in microglia or astrocytes as determined by immunohistochemistry and confocal microscopy. However, the increased IL-6-immunoreactivity was mainly expressed in blood vessels. The differential upregulation of TNF-α and IL-6 induced by incision suggests that the proinflammatory cytokines may play different roles in the development of surgical pain.

To observe the expression distribution of bone morphogenetic proteins (BMP) in the spinal cord of normal adult rats.Expression of BMP2, BMP4, and BMP7, and their receptors BMPR Ia, BMPR Ib, and BMP II were detected by immunochemistry analysis in the spinal cord of normal adult rats.Expression of BMPR Ia or BMPR Ib was observed in the motor neurons of the anterior horn, sensory neurons of the dorsal horn, oligodentrocytes, some microglia, and some astrocytes. Expression of receptor BMPR II was found in the oligodentrocytes and motor neurons in the gray matter of anterior horn. It was also expressed in some glial fibrillary acidic protein (GFAP)-positive astrocytes in the white matter but not in the gray matter. BMP2 and BMP4 were not expressed in the spinal cord of normal adult rats by immunohistochemistry. BMP7 was expressed in all the APC-positive oligodentrocytes, all the NeuN-positive motor neurons in the anterior horn, and some astrocytes in the normal spinal cord. Phosphated pSmad 1/5/8 protein was expressed in all the oligodentrocytes, all the neurons, and some astrocytes, especially in the GFAP-positive astrocytes which were RC2-positive radial glia in the subventricular zone.BMP7, BMP receptors, and phosphated pSmad 1/5/8 are expressed in many types of cells whereas BMP2 and BMP4 are not expressed in the spinal cord of normal adult rats, which suggests an important function of BMP signal pathway in the neuron and glia of spinal cord.

To investigate the effect of morphine preconditioning on mitochondrial permeability transition pore (MPTP) and its protective mechanism after myocardial ischemia-reperfusion injury.A rat model of ischemia-reperfusion injury was established. Forty rats were injected with 2-(3)[H] DOG and then divided into 4 groups randomly: a sham operation (S) group, an ischemia-reperfusion injury (IR) group, a morphine preconditioning (Mp+IR) group, and a cyclosporine A preconditioning (CsA+IR) group. We monitored the concentrations of serum creatine kinase-Mb (CK-Mb) and cardiac troponin I (cTnI), and measured myocardial mitochondrial 2-(3)[H] DOG, cytochrome c content, Ca(2+) concentration ([Ca(2+)]m), the velocity of Ca(2+) intake and reaction half time of mitochondrial permeability transition pore (MPTP t(1/2)) in the 4 groups.The concentrations of serum CK-Mb and cTnI decreased more in the Mp+IR group and the CsA+IR group than those of the IR group. The concentrations of 2-(3)[H]DOG and [Ca(2+)]m in the IR group were evidently higher but the level of cytochrome c was lower than those of the sham operation group. The concentrations of 2-(3)[H] DOG and [Ca(2+)]m in the Mp+IR group decreased whereas the concentration of cytochrome c increased compared with those in the IR group. Mitochondrial 2-(3)[H]DOG content was positively correlated with the concentration of calcium (r=0.797, P0.01). The 2-(3)[H]DOG and [Ca(2+)]m content were negatively correlated with cytochrome c in the IR group (r=-0.805 and r=-0.648, respectively, P0.01). MPTP t(1/2) in the IR group was shortened evidently, and that in the Mp+IR and CsA+IR group was significantly lengthened.Morphine preconditioning may have myocardial protective effect through unburdening the calcium overload and lengthening the MPTP t(1/2).

To explore the role of phosphorylation of cAMP response element binding protein (CREB) in the incision-induced pain hypersensitivity.A longitudinal incision was made in one plantar hind paw of isoflurane-anesthetized rats. Spinal cords were removed at various postoperative time after behavior test. Phosphorylation of CREB was determined by immunohistochemistry and double-labeling immunofluorescence. Morphine and gabapentin were intraperitoneally injected before the behavior test and were used to determine the interaction between phosphorylation of CREB and morphine and gabapentin.After the hind-paw incision, phosphorylation of CREB was enhanced in the ipsilateral lumbar spinal cord (P0.05). The enhancement of p-CREB was mainly in the neurons in the dorsal horn of the spinal cord. All these were shown by double-labeling technique and p-CREB was mainly in the neurons. Intraperitoneal injection of morphine prevented the increased phosphorylation of CREB in the spinal cord and inhibited the mechanical allodynia induced by the incision (P0.05). Gabapentin didn't inhibit the phosphorylation of CREB (P0.05) but partly inhibited the mechanical allodynia.Incision induces the phosphorylation of CREB in the spinal cord, and the increase of p-CREB is mainly in the neurons. Phosphorylation of CREB in the spinal cord contributes to the pain hypersensitivity induced by surgical incision.

To investigate the effect of acute cardiac injury on the activation of interleukin-1 beta (IL-1 beta) signaling in the spinal cord.Acute cardiac injury rat model was established by intra-myocardial injection of formalin through diaphragm. IL-1 beta expression was determined by Western blot, immunohistochemistry and in situ hybridization. The DNA binding activities of 2 IL-1 beta transcription factors, activator protein (AP)-1 and nuclear factor kB (NF-kappaB) were measured by electrophoretic mobility shift assay (EMSA).After cardiac injury, the IL-1 beta protein level was dramatically upregulated in the spinal cord. The upregulated IL-1 beta was mainly expressed in the neurons in the lamina II approximately IV of the spinal cord. In response to cardiac injury, the DNA binding activities of NF-kappaB and AP-1 were greatly activated.Acute cardiac injury could activate the spinal IL-1 beta signaling, which, in turn, may be involved in the progression of heart failure after injury.

To explore the effect of isoflurane preconditioning on the plasma concentration of matrix metalloproteinase (MMP)-9 and myocardial ultramicrostructure in patients undergoing cardiac valve replacement.Thirty patients undergoing elective cardiac valve replacement with cardiopulmonary bypass (CPB) were randomly assigned to a control group (ný15) and an isoflurane group (ný15). In the isoflurane group, isoflurane of 1.0 minimum alveolar concentration end-tidal( 1.1% approximately 1.2%) was administered for 30 min followed by a 15 min washout period before the CPB. The control group did not inhale isofurane, and there was no difference in the other drugs in the 2 groups. Blood samples for MMP-9 were obtained before incision(T(0)) and at 30 min (T(1)),6 h (T(2)),12 h (T(3)), and 24 h (T(4)) after the reperfusion. Right atrial biopsies were collected before and after the CPB to observe the myocardial ultramicrostructure.Compared with T(0), the mean MMP-9 level significantly increased at T(1), T(2) and T(3) in the control group(P0.01), while the MMP-9 level only at T(1) significantly increased in the isoflurane group (P0.01). The mean MMP-9 level was significantly reduced in the isoflurane group at T(2) compared with each time point in the control group. The difference in MMP-9 levels between T(1), T(2), T(3) and T(0) was significantly lower in the isoflurane group than that in the control group (P0.01). The ultramicrostructure injury of myocardium under electron microscope in the control group was worse than that in the isoflurane group.The plasma concentration of MMP-9 is inhibited by isoflurane preconditioning in patients undergoing cardiac valve replacement after CPB, which might be part of its protective mechanism against myocardium injury after CPB.

Background: The pathogenic role of brain derived neurotrophic factor (BDNF) in the incisional pain is poorly understood. The present study explores the role of the BDNF in the incision-induced pain hypersensitivity. Methods: A longitudinal incision was made in one plantar hind paw of isoflurane-anesthetized rats. Dorsal root ganglias (DRG) and spinal cords were removed at various postoperative times (1–72 h). Expression pattern of BDNF was determined by immunohistochemistry and double-labeling immunofluorescence. Lidocaine-induced blockade of sciatic nerve function was used to determine the importance of afferent nerve activity on BDNF expression in the DRG and spinal cord after incision. BDNF antibody was administered intrathecally (IT) or intraperitoneal (IP) to modulate the spinal BDNF or peripheral BDNF after incision. Results: After hind-paw incision, the BDNF was upregulated in the ipsilateral lumbar DRG and spinal cord whereas thoracic BDNF remained unchanged in response to incision. The upregulated BDNF was mainly expressed in the large-sized neurons in DRG and the neurons and the primary nerve terminals in the spinal cord. Sciatic nerve blockade prevented the increase of BDNF in the DRG and spinal cord. IT injection of BDNF antibody greatly inhibited the mechanical allodynia induced by incision whereas IP administration had only marginal effect. Conclusion: The present study showed that incision induced the segmental upregulation of BDNF in the DRG and spinal cord through somatic afferent nerve transmission, and the upregulated BDNF contributed to the pain hypersensitivity induced by surgical incision.

Previous studies have shown that microglia impact the proliferation and differentiation of neurons during hippocampal neurogenesis via the fractalkine/CX3 chemokine receptor 1 (CX3CR1) signaling pathway. However, whether microglia can influence the maturation and dendritic growth of newborn neurons during hippocampal neurogenesis remains unclear. In the present study, we found that the number of doublecortin-positive cells in the hippocampus was decreased, and the dendritic length and number of intersections in newborn neurons in the hippocampus were reduced in transgenic adult mice with CX3CR1 deficiency (CX3CR1 (GFP/GFP) ). Furthermore, after experimental seizures were induced with kainic acid in these CX3CR1-deficient mice, the expression of c-fos, a marker of neuronal activity, was reduced compared with wild-type mice. Collectively, the experimental findings indicate that the functional maturation of newborn neurons during hippocampal neurogenesis in adult mice is delayed by CX3CR1 deficiency.