Identification of brain-to-spinal circuits controlling the laterality and duration of mechanical allodynia in mice.

Mechanical allodynia (MA) represents one prevalent symptom of chronic pain. Previously we and others have identified spinal and brain circuits that transmit or modulate the initial establishment of MA. However, brain-derived descending pathways that control the laterality and duration of MA are still poorly understood. Here we report that the contralateral brain-to-spinal circuits, from Oprm1 neurons in the lateral parabrachial nucleus (lPBNOprm1), via Pdyn neurons in the dorsal medial regions of hypothalamus (dmHPdyn), to the spinal dorsal horn (SDH), act to prevent nerve injury from inducing contralateral MA and reduce the duration of bilateral MA induced by capsaicin. Ablating/silencing dmH-projecting lPBNOprm1 neurons or SDH-projecting dmHPdyn neurons, deleting Dyn peptide from dmH, or blocking spinal κ-opioid receptors all led to long-lasting bilateral MA. Conversely, activation of dmHPdyn neurons or their axonal terminals in SDH can suppress sustained bilateral MA induced by lPBN lesion.

Effect of meridian sinew releasing technique on moxibustion sensation of heat-sensitive moxibustion in patients with knee osteoarthritis / 中国针灸

OBJECTIVE@#To observe the effect of meridian sinew releasing technique on moxibustion sensation of heat-sensitive moxibustion in patients with knee osteoarthritis (KOA).@*METHODS@#A total of 60 patients with KOA were randomly divided into an observation group and a control group, 30 cases each group. In the observation group, on the basis of the meridian sinew releasing technique, moxibustion sensation exploration method was applied at Dubi (ST 35) area on the affected side. In the control group, moxibustion sensation exploration method was applied at Dubi (ST 35) area on the affected side. The meridian sinew releasing technique was performed for 20 min each time, the moxibustion sensation exploration method was performed for 60 min each time, once a day for 3 days. The excitation rate, latency, duration time and intensity value of moxibustion sensation of heat-sensitive moxibustion were recorded on the 1st, 2nd and 3rd days of exploration in the two groups.@*RESULTS@#The excitation rate on the 3rd day of exploration and total excitation rate in the observation group were higher than the control group (P<0.05). On the 1st, 2nd and 3rd days of exploration, the latency of moxibustion sensation of heat-sensitive moxibustion in the observation group was shorter than the control group (P<0.05), the duration time was longer than the control group (P<0.05), and the intensity value was higher than the control group (P<0.05).@*CONCLUSION@#Meridian sinew releasing technique could improve the excitation rate of moxibustion sensation of heat-sensitive moxibustion in patients with KOA, shorten the latency, prolong the duration time, and improve the intensity value.

Salidroside Ameliorates Ultraviolet-Induced Keratinocyte Injury by Inducing SIRT1-Dependent Autophagy.

Introduction: Autophagy is an important process for maintaining intracellular homeostasis and is deregulated in ultraviolet B (UVB)-induced skin injury. Salidroside (SAL) is an active ingredient extracted from Rhodiola rosea, which is a herbal medicine that has shown protection against ultraviolet (UV) radiation. Here, we investigated the functions and mechanisms of SAL on UVB-induced skin cell oxidative damage and autophagy. Methods: Human immortalized keratinocyte cell line HaCaT was used as a cell model of UV injury. HaCaT cells were exposed to UVB irradiation and then incubated with SAL to investigate cell viability, lactate dehydrogenase (LSD) in culture media, intracellular reactive oxygen species (ROS) level, oxidative stress, autophagy, and regulatory effects on SIRT1 protein. Results: SAL pretreatment (25, 50 and 100 µM) increased cell viability and inhibited LDH release in UVB-challenged HaCaT cells. SAL (100 µM) significantly reduced intracellular ROS level and suppressed oxidative stress, with increased MDA content and increased SOD activity. In addition, SAL pretreatment enhanced autophagy in UVB-irradiated HaCaT cells, increased protein expressions of Beclin-1 and ATG7, and decreased protein expression of P62. We also found that pretreatment with SAL increased the SIRT1 protein in irradiated HaCaT cells. SAL protected UVB-induced damage in a dependent manner on autophagy and SIRT1, as SAL-induced increase in viability was significantly attenuated by specific autophagy inhibitor Wortmannin (1 µM) or SIRT1 inhibitor EX-527 (100 nM). Discussion: The present study results speculate that SAL suppresses UVB-induced injury and autophagy by enhancing SIRT1 expression.

Clinical guidelines for microwave ablation of spinal metastases.

Spinal metastases are the most common source of morbidity in patients with cancer. Recently, microwave ablation has produced satisfactory results in the management of spinal metastases. However, there is still controversy in terms of clinical treatment, such as indication, power, time, and temperature. To standardize the application of microwave ablation technology and reduce the risk of surgical-related complications in spinal metastases, in this report, we aimed to summarize the current evidence and clinical experience of microwave ablation and developed a clinical guideline, initiated by the Musculoskeletal Tumor Group of the Committee for Minimally Invasive Therapy in Oncology of the Chinese Anti-Cancer Association. The Grading of Recommendations Assessment, Development, and Evaluation (GRADE) approach was used in to rate the quality of evidence and the strength of recommendations, and the Reporting Items for Practice Guidelines in Healthcare (RIGHT) checklist was strictly followed to report the guideline. Finally, 15 evidence-based recommendations were formulated based on the 15 most concerned clinical questions among orthopedic surgeons, oncologists, and interventional radiologists in China. This guideline aims to promote the science-based normalization of microwave ablation for the treatment of spinal metastases.

Astragalus polysaccharide alleviates cognitive impairment and ß-amyloid accumulation in APP/PS1 mice via Nrf2 pathway.

Alzheimer's disease (AD) is the most common neurodegenerative disorder, and its etiology and pathogenesis are not fully understood. Astragalus polysaccharide (APS) has many pharmacological activities, but there are few reports about its role in AD. Using the common AD model APP/PS1 mice, it was found that the expression of Keap1 (a negative regulatory factor of Nrf2), the protein level of cytoplasmic Nrf2 and the content of MDA were increased significantly, while the mRNA level of Nrf2, the expression of Nrf2 in nucleus and the contents of SOD and GSH-Px were decreased significantly. APS treatment significantly increased the expression of Nrf2 in the nucleus but decreased its expression in the cytoplasm, and restored the expression levels of Keap1, SOD, GSH-Px and MDA. When APP/PS1 mice were treated with APS and injected with Nrf2 siRNA, the down-regulation of Nrf2 expression significantly blocked the regulation of APS on oxidative stress. Continuing to test the physiological function of AD mice showed that the spatial learning and memory abilities of APP/PS1 mice were impaired, the apoptosis of brain cells and the content of ß-amyloid (Aß) were significantly increased. APS treatment significantly improved the cognitive ability of APP/PS1 mice, reduced apoptosis and the accumulation of Aß, but the above effects of APS were blocked by Nrf2 siRNA injection. Therefore, APS can activate Nrf2 pathway to improve the physiological function of AD mice, which may have important clinical application value.

[Identification of chemical constituents in Guizhi Fuling Capsules by UPLC-Q-TOF-MS/MS].

To qualitatively characterize the chemical composition of Guizhi Fuling Capsules using UPLC-ESI-Q-TOF-MS/MS. The analysis was performed on Agilent ZORBAX RRHD Eclipes Plus C_(18)(2.1 mm×100 mm, 1.8 µm) column,that was eluted with mobile phase consisting of acetonitrile and 0.1% formic acid in a gradient mode. The flow rate was 0.4 mL·min~(-1), and column temperature was 30 ℃. Tandem mass spectrometry was acquired in both negative and positive ESI modes. These components were further analyzed based on high-resolution mass-to-charge ratios, fragment ion species, reference substances and literature data. In conclusion, a total of 200 compounds were identified, in which 40 were verified with reference substances. The current study laid a foundation for in-depth studies of its mass balance and pharmacodynamics.

Vasodilatory effect and structural-activity relationship of a group of iridoid glucosides from Phlomis likiangensis.

As a folk medicine, Phlomis likiangensis is traditionally used in China to activate collaterals and protect cardiovascular system. We hypothesized that the beneficial effects of Phlomis likiangensis may be related to vasodilatation. In the present study, twelve known iridoid glucosides (1-12) were isolated from Phlomis likiangensis. The vasodilatory effects and the underlying mechanisms of the main components (iridoid glucosides) of Phlomis likiangensis on rat aortic rings were investigated. The result showed that iridoid glucosides significantly increased the vasodilatation in rat aortic rings, which was abolished by removing the endothelium of the vessels or by eliminating the generation of nitric oxide. Finally, the structure-activity relationship of compounds 1-12 was also speculated. Our findings provide the first evidence that the iridoid glucosides of Phlomis likiangensis may be the pharmacodynamic basis for its traditional efficacy.

Nanotherapy in Joints: Increasing Endogenous Hyaluronan Production by Delivering Hyaluronan Synthase 2.

Osteoarthritis (OA) is a common joint degenerative disease that causes pain, joint damage, and dysfunction. External hyaluronic acid (HA) supplement is a common method for the management of osteoarthritis which requires multi-injections. It is demonstrated that biodegradable mesoporous silica nanoparticles successfully deliver an enzyme, hyaluronan synthase type 2 (HAS2), into synoviocytes from the temporomandibular joint (TMJ) and generate endogenous HA with high molecular weights. In a rat TMJ osteoarthritis inflammation model, this strategy promotes endogenous HA production and inhibits the synovial inflammation of OA for more than 3 weeks with one-shot administration. Such nanotherapy also helps repairing the bone defects in a rat OA bone defect model.

Persistent high-energy spin excitations in iron-pnictide superconductors.

Motivated by the premise that superconductivity in iron-based superconductors is unconventional and mediated by spin fluctuations, an intense research effort has been focused on characterizing the spin-excitation spectrum in the magnetically ordered parent phases of the Fe pnictides and chalcogenides. For these undoped materials, it is well established that the spin-excitation spectrum consists of sharp, highly dispersive magnons. The fate of these high-energy magnetic modes upon sizable doping with holes is hitherto unresolved. Here we demonstrate, using resonant inelastic X-ray scattering, that optimally hole-doped superconducting Ba(0.6)K(0.4)Fe(2)As(2) retains well-defined, dispersive high-energy modes of magnetic origin. These paramagnon modes are softer than, though as intense as, the magnons of undoped antiferromagnetic BaFe(2)As(2). The persistence of spin excitations well into the superconducting phase suggests that the spin fluctuations in Fe-pnictide superconductors originate from a distinctly correlated spin state. This connects Fe pnictides to cuprates, for which, in spite of fundamental electronic structure differences, similar paramagnons are present.

Effects of extract of ginkgo biloba on learning and memory ability and NGF and NT-3 expression in diabetic rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effect of extract of Ginkgo Biloba(EGB) on nerve growth factor(NGF) and Neurotrophin-3(NT-3) expression of hippocampus neurons in streptozotocin-induced type I diabetic rats.</p><p><b>METHODS</b>Thirty male SD rats were divided into three groups (n = 10): the control group, diabetic group and EGB-treated group. Strepozotocin were injected intraperitoneally in the later two groups to induce diabetes. EGB-treated group was injected intraperitoneally with EGB, and the same volume of normal saline was injected to the other groups. Concentration of blood glucose and body weight and behaviour were dynamicly monitored. At the end of the 12th week, morphological changes of the hippocampus neurons were observed under microscopy by HE stain. The expression of NGF and NT-3 were assayed by Western blot and RT-PCR respectively.</p><p><b>RESULTS</b>Compared with diabetic group, the behaviour and body weight (P < 0.05) and the concentration of blood glucose (P < 0.05) were significantly improved and the escape latency of Morris water maze test (P < 0.05) was significantly shortened, while the platform searching score was significantly increased (P < 0.01) in EGB treated group; The pathological changes of hippocampus neurons were significantly attenuate by EGB treated; The expression of NGF and NT-3 in hippocampus neurons were significantly increased which assayed by Western blotting and RT-PCR respectively (P < 0.05) in EGB treated group.</p><p><b>CONCLUSION</b>EGB may improve the learning and memory ability of diabetic rats the mechanism may be attributed to its improvement of the expression of NGF and NT-3 and reducing apoptosis in hippocampus neurons.</p>

Role of P38-MAPK signal transduction pathway and effect of panax notoginoside in rats with hypoxic hypercapnia pulmonary hypertension / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the role and significance of P38-MAPK in the pathological process of hypoxic hypercapnia pulmonary hypertension in rats, and the protection of panax notoginoside (PNS).</p><p><b>METHODS</b>(1) To set up rat pathological model of hypoxic hypercapnia pulmonary hypertension: seventy two male SD rats (200 280 g) were randomly divided into six groups (n = 12), which were normal group (N group), hypoxic hypercapnia for 3-day group (H3d), hypoxic hypercapnia for 1-week group(H1w), hypoxic hypercapnia for 2-week group (H2w), hypoxic hypercapnia for 4-week group (H4w) and PNS-injected group (Hp). The rats of PNS -injected group were injected PNS before being placed in the chamber (50 mg/(kg x d), ip), and other groups were injected normal sodium (2 ml/kg, ip). (2) The shapes of pulmonary artery were detected by HE staining. (3) Western blot was used to study the protein expression of p38-MAPK. The expression of p38-MAPK in lung tissue and pulmonary blood vessel was investigated by immunohistochemistry.</p><p><b>RESULTS</b>(1) The ratio of vessel wall area/total area (WA/ TA) in H1w, H2w, H4w and Hp group was higher than that of N group (P < 0.05), but that of H3d group did not change obviously (P > 0. 05 vs N group). The ratio of WA/TA in Hp group was obviously lower than that of H4w, group (P < 0.05). (2) The levels of P-p38 protein was markedly ascended in H3d group (0.225 +/- 0.071) compared with N group (0.012 +/- 0.006), and expression of P-p38 protein was significantly positive in H1w, H2w, H4w groups. (P < 0.05). (3) As P-p38 protein in pulmonary arterial tunica intima and tunica media, sterile expression in N group (0.099 +/- 0.015) and H3d group (0.107 +/- 0.013) contrasted to H4w group (0.124 +/- 0.025, P < 0.05), then tended to rise in H2w, H4w group (P < 0.05). (4) In pulmonary tissue, the levels of P-p38 protein in PNS-injected group were lower 53.02% (P < 0.05) than those in H4w group. In pulmonary arterial tunica intima and tunica media the levels of P-p38 protein in PNS-injected group were lower 87.33% (P < 0.05) than those in H4w group.</p><p><b>CONCLUSION</b>p38-MAPK as a signal transduction may play an important role in the development of hypoxia induced pulmonary hypertension. The effect of PNS on reducing pulmonary hypertension and improving pulmonary vascular wall remodeling may be related to its inhibiting expression of p38 MAPK.</p>

Effects of astragalus and angelica on bone marrow stem cells proliferation and VEGF protein expression in vitro / 中国骨伤

<p><b>OBJECTIVE</b>To observe the effects of Astragalus and Angelica on bone marrow stem cells (BMSU) proliteratlon mn vitro and investigate its possible mechanism.</p><p><b>METHODS</b>Five 200 to 220 g SD rats were fed with a high fat diet for 4 weeks and given 30 mg/kg streptozotocin (STZ) twice develop type II diabetes from July 2009 to February 2010. The rats with blood glucose concentrations of 16.7 mmol/L or more were considered diabetic. Bone Marrow Stem Cells (BMSC) were collected and isolated by density gradient centrifugation. The BMSC were divided into 4 groups,including empty control group, Astragalus group, Angelica group and Astragalus plus Angelica group. DMEM of 100 microl was added in empty control group. DMEM of 100 microl containing Astragalus (1100 mg/L), Angelica (1100 mg/L) and Astragalus (1100 mg/L) combine with Angelica(220 mg/L) were added in Astragalus group, Angelica group and Astragalus plus Angelica group respectively. The cell proliferation was detected by MTT method, and the concentration of VEGF in the supernatant was determined by ELISA. The VEGF expression was analyzed by Western Blot after 14 days incubation.</p><p><b>RESULTS</b>The BMSC proliferation and the VEGF concentration in the supernatant and the BMSC VEGF protein expression significantly increased in Astragalus group and Astragalus plus Angelica group compared to those of empty control group (P < 0.05 or P < 0.01). The above effects were more strong in Astragalus plus Angelica group (P < 0.05).</p><p><b>CONCLUSION</b>Astragalus with Angelica or used separately could promote BMSC proliferation. The mechanism might induce the VEGF protein expression in BMSC. And the independent use of Angelica has no above effect.</p>

[Preventive and treatment effect of composite Rhodiolae on acute lung injury in patients with severe pulmonary hypertension during extracorporeal circulation].

OBJECTIVE: To investigate the possibility of early prevention and treatment of acute lung injury by using composite Rhodiolae (CR) in patients with severe pulmonary hypertension during extracorporeal circulation. METHODS: Seventy-six patients with severe pulmonary hypertension screened out by color 2D-Doppler ultrasonography were randomly divided into the treated group (n = 40) and the control group (n = 36). The general treatment for the two groups, including preoperational preparation, operational procedure and conventional drug therapy, was the same. To the treated group, CR was administered 4 g every day (half the dose in children) at 7-10 day before and 5-7 day after the operation. The life signs, pulmonary ventilation parameters, arterial blood gas analysis were monitored and blood levels of TXB2, 6-keto-PGF1 alpha in both groups were determined in the fixed time points. RESULTS: The various parameters in the treated group were all better than those in the control group (P < 0.05 or P < 0.01), the occurrence rate of acute lung injury and its mortality in the treated group were 7.5% (3/40) and 0% (0/3) respectively, while in the control group were 19.4% (7/36) and 43% (3/7) respectively. CONCLUSION: CR has good preventive and treatment effect in treating complicated acute lung injury during extracorporeal circulation (cardiopulmonary bypass) in patients with pulmonary hypertension.

[Enhancement of cellular immune response to DNA vaccine encoding hepatitis C virus core and envelope 2 fusion antigen by murine Fms-like tyrosine kinase 3 ligand].

Hepatitis C virus (HCV) is an important human pathogen that causes chronic liver disease worldwide. It is desirable to develop vaccines to prevent HCV infection, or at least to prevent progression to chronicity. We once constructed an optimized hepatitis C virus core and envelope 2 fusion antigen DNA vaccine, which could induce humoral and cellular immune responses against HCV core and E2 protein in BALB/c mice efficiently. Flt3 (Fms-like tyrosine kinase 3) -ligand has been identified as an important cytokine for the generation of professional antigen-presenting cells, particularly dendritic cells. We reasoned that a DNA vaccine coexpressing the antigen and FL may activate immune responses more effectually. In this study, The influence of FL on this HCV DNA vaccine was evaluated. The cDNA encoding signal peptide and extracellular domain of murine FL was inserted into the plasmid pST-CE2t, and the resulting plasmid pST-CE2t/FL was transfected into COS7 cells. The HCV core and E2 protein were detected by Western blotting, and the soluble murine FL was detected by ELISA. Eight-week-old female BALB/c mice were inoculated intramuscularly with 100 microg pST-CE2t, pST-CE2t/FL or mock vector, respectively, and boosted at the same dosage 3 weeks later. Anti-HCV core and E2 total IgG and isotypes were measured at weeks 1,3,5,7. Splenocyte proliferative response to recombinant HCV core and E2 protrein were detected at week 7. SP2/0 cells expressing HCV core protein were used as target cells for the detection of cytotoxic T lymphocyte (CTL) response. Western blot analysis showed that a protein band with molecular weight about 70 kD from lysate of COS7 cells transfected with plasmid pST-CE2t/FL could be detected by anti-HCV core or E2 monoclonal antibodies, which indicated that pST-CE2t could express glucosylated HCV core and E2 fusion protein. Murine FL could be detected in the culture supernatant of COS7 cells transfected with pST-CE2t/FL. Plasmid pST-CE2t immunized mice developed higher anti-HCV core and E2 IgG seroconversion rates and titers than pST-CE2t/FL group did at different various times, but the IgG2a/IgG1 ratio of anti-HCV E2 protein in pST-CE2t/FL group is much higher than pST-CE2t group. Splenocytes from pST-CE2t or pST-CE2t/FL immunized mice could proliferate with stimulation of HCV core or E2 protein in vitro, although pST-CE2t/FL group showed much stronger response. Splenocytes from mice immunized with pST-CE2t/FL induced 79.03% +/- 9.95% of target cell lysis at the effector/target ratio of 100:1, which was significantly greater than the lysis (62.2% +/- 8.62%) observed in mice immunized with pST-CE2t. Our data demonstrated that the incorporation of FL can preferentially enhance the cellular response to this HCV fusion antigen DNA vaccine. In contrast, HCV specific antibodies were inhibited by FL in vaccinated mice. More and more data supports that recovery from acute HCV infection may depend upon the generation of broad-based cellular immune responses to viral proteins. So, FL may be of potential value as an adjuvant in the development of DNA-based immunization for prophylactic and therapeutic vaccine against HCV infection.

Preventive and treatment effect of composite Rhodiolae on acute lung injury in patients with severe pulmonary hypertension during extracorporeal circulation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the possibility of early prevention and treatment of acute lung injury by using composite Rhodiolae (CR) in patients with severe pulmonary hypertension during extracorporeal circulation.</p><p><b>METHODS</b>Seventy-six patients with severe pulmonary hypertension screened out by color 2D-Doppler ultrasonography were randomly divided into the treated group (n = 40) and the control group (n = 36). The general treatment for the two groups, including preoperational preparation, operational procedure and conventional drug therapy, was the same. To the treated group, CR was administered 4 g every day (half the dose in children) at 7-10 day before and 5-7 day after the operation. The life signs, pulmonary ventilation parameters, arterial blood gas analysis were monitored and blood levels of TXB2, 6-keto-PGF1 alpha in both groups were determined in the fixed time points.</p><p><b>RESULTS</b>The various parameters in the treated group were all better than those in the control group (P < 0.05 or P < 0.01), the occurrence rate of acute lung injury and its mortality in the treated group were 7.5% (3/40) and 0% (0/3) respectively, while in the control group were 19.4% (7/36) and 43% (3/7) respectively.</p><p><b>CONCLUSION</b>CR has good preventive and treatment effect in treating complicated acute lung injury during extracorporeal circulation (cardiopulmonary bypass) in patients with pulmonary hypertension.</p>