The Tongmai Yangxin pill (TMYX) has potential clinical effects on no-reflow (NR); however, the effective substances and mechanisms by which this occurs remain unclear. This study evaluates the cardioprotective effects and molecular mechanisms of TMYX against NR. We used a myocardial NR rat model (2 h after myocardial ischemia and 2 h after reperfusion) to confirm the effect and mechanism of action of TMYX in alleviating NR. In vitro studies in isolated coronary microvasculature of NR rats and in silico network pharmacology analyses were performed to reveal the underlying mechanisms of TMYX and determine the main components, targets, and pathways of TMYX, respectively. The experiment was approved by the Ethics Committee of Hunan University of Chinese Medicine (LLBH-202212160001). TMYX showed therapeutic effects on NR by improving cardiac structure and function, reducing NR, ischemic areas, and cardiomyocyte injury, and decreasing the content of cardiac troponin I (cTnI). Moreover, the mechanism of TMYX predicted by network pharmacology is related to the hypoxia inducible factor-1 (HIF-1), nuclear factor kappa-B (NF-κB), and tumor necrosis factor (TNF) signaling pathways. TMYX increased the expression of G protein-coupled estrogen receptor (GPER), phospho-extracellular signal-regulated kinase (p-ERK), and HIF-1α. In vitro, TMYX enhanced the diastolic function of coronary microvascular cells; however, this effect was inhibited by GPER inhibitor (G-15), eNOS inhibitor (L-NAME), and sGC inhibitor (ODQ). This study integrates pharmacology and experimental evaluation to reveal that TMYX activates HIF-1α/eNOS signaling pathway by upregulating GPER to relax coronary microvessels, thereby significantly alleviating NR.

Objective To study the relationship between activity of thorax and each spinal intervertebral angle in patients with ankylosing spondylitis. Methods Each spinal intervertebral angle of 41 patients with ankylosing spondylitis were measured by Spinalmouse in different postures. And the activity of thorax was measured. Correlation between activity of thorax and shape of spinal were analyzed. Results The activity of thorax positively correlated with the entire lumbar spinal column in flexion (P<0.01), as well as the intervertebral angle of L1/2, L2/3 and L4/5 in flexion (P<0.05), but negatively correlated with the intervertebral angle of L1/2 and L2/3, curvature of the entire lumbar spinal column in upright and the intervertebral angle of L1/2, L3/4, curvature of the entire lumbar spinal column in extension. Conclusion There was a significant relation between activity of thorax and lumbar vertebra motor ability.

A sensitive and selective liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed and validated for the determination of betamethasone in human plasma. The analyte was isocratically eluted on a Venusil XBP C8 column (200 mm x 3.9 mm ID, 5 microm) with methanol-water mol x L(-1) ammonium formate) (80:20) at a flow rate of 0.4 mL x min(-1), and detected (containing 5 mmol x L(-1) ammonium formate) (80:20) at a flow rate of 0.4 mL x min(-1), and detected with a triple quad LC-MS/MS using ESI with positive ionization. Ions monitored in the multiple reaction monitoring (MRM) mode were m/z 393.3-->355.2 for betamethasone and m/z 361.3-->343.2 for prednisolone (IS). Betamethasone was extracted from 0.5 mL human plasma with ethyl acetate. The average recovery is 88.24% and the low limit of quantitation (LLOQ) was 0.5 ng x mL(-1). The 3-day validation study demonstrated excellent precision and accuracy across the calibration range of 0.5-80.0 ng x mL(-1). The method was successfully applied to the pharmacokinetic study of compound betamethason injection in healthy Chinese volunteers.
Anti-Inflammatory Agents ; blood ; pharmacokinetics ; Area Under Curve ; Betamethasone ; blood ; pharmacokinetics ; Chromatography, Liquid ; methods ; Humans ; Injections, Intramuscular ; Male ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry ; methods ; Young Adult

OBJECTIVE: To evaluate the effects of madecassoside (MC) on the depression behavior of mice and the activities of monoamine oxidase (MAO) in different rat brain regions. METHODS: Imipramine as the positive contrast medicine, effects of MC on the depression behavior of mice were observed by forced swimming test and reserpine antagonist test. Moclobemide and pargyline as the positive controlled medicines, the activities of monoamine oxidase-A (MAO-A) and monoamine oxidase-B (MAO-B) in different rat brain regions were determined after intragastric administration of MC in 3 different dosages for 3 days or 21 days. RESULTS: (1) The low, middle and high dosages of MC (i.g.) significantly reduced the immobility time of mice in forced swimming test (P<0.05). (2) MC in dosages of 10 mg/kg and 20 mg/kg prevented the lowering of temperature induced by reserpine (P<0.05), while 40 mg/kg had no significant effects on it (P>0.05). (3) With acute administration (3 days), the low, middle and high dosagey of MC (i.g.) significantly inhibited the activity of MAO-A in hippocampus (P<0.01), and the high dosage significantly inhibited the activity of MAO-A in hypothalamus (P<0.01), while the 3 dosages had no significant effects on the activity of MAO-A in cortex (P>0.05). With chronic administration (21 days), MC in 3 dosages had no significant effects on the activities of MAO-A in cortex and hypothalamus (P>0.05), and the high dosage (40 mg/kg) significantly enhanced the activity of MAO-A in hippocampus (P<0.01). (4) With acute administration, MC in dosages of 10 mg/kg and 20 mg/kg significantly inhibited the activity of MAO-B in cortex (P>0.05), and MC in dosage of 10 mg/kg significantly inhibited the activity of MAO-B in hypothalamus (P<0.05), and MC in dosage of 20 mg/kg significantly enhanced the activity of MAO-B in hippocampus (P<0.01). With chronic administration, MC of 3 dosages produced no significant effects on the activities of MAO-B in 3 different rat brain regions (P>0.05). CONCLUSION: These results support the idea that MC produces antidepressant effects through MAO inhibition in rat brain, which seems stronger with acute administration than chronic administration, while its mechanism remains to be further studied.

Objective To develop an accurate,reliable method without the need of initialization in animal PET modeling for estimation of the tracer kinetic parameters based on the artificial immune network.Methods The hepatic and left ventricular time activity curves (TACs) were obtained by drawing ROIs of liver tissue and left ventricle on dynamic 18F-FDG PET imaging of small mice.Meanwhile,the blood TAC was analyzed by sampling the tail vein blood at different time points after injection.The artificial immune network for parametric optimization of pharmacokinetics (PKAIN) was adapted to estimate the model parameters and the metabolic rate of glucose (Ki) was calculated.Results TACs of liver,left ventricle and tail vein blood were obtained.Based on the artificial immune network,Ki in 3 mice was estimated as 0.0024,0.0417 and 0.0047,respectively.The average weighted residual sum of squares of the output model generated by PKAIN was less than 0.0745 with a maximum standard deviation of 0.0084,which indicated that the proposed PKAIN method can provide accurate and reliable parametric estimation.Conclusion The PKAIN method could provide accurate and reliable tracer kinetic modeling in animal PET imaging without the need of initialization of model parameters.

Objective:To investigate the effect of dihydromyricetin (DMY) on LPS-induced septic shock in mice and the related underlying mechanism.Methods:The LPS-induced septic shock mice model was established after the mice were pre-treated by DMY for 7 days.The mortality rate was calculated at 24,48,72,96,120,144 and 168 h after the mice were intraperitoneal injected with LPS.For elucidation of underlying mechanism ,RAW246.7 were pre-incubated with DMY for 1 h,and then stimulated by LPS 100 ng/ml.Western blot was performed for determination of P-ERK,P-JNK and P-p38 expression.Immunohistochemistry was applied to explore c-Fos and c-Jun nucleus translocation.Results:DMY could significantly inhibit LPS-induced mice mortality.Inhibitory effect of DMY on the phosphorylation of JNK and p 38 contributed to the anti-inflammatory effect of DMY in vivo.Furthermore , DMY obviously prevented c-Fos and c-Jun nucleus translocation.Conclusion:The anti-inflammatory effect of DMY is attributed to the suppression on c-Fos and c-Jun nucleus translocation ,via inhibition of the phosphorylation of JNK and p 38.

Objective To investigate the relations and evaluation value of serum thymidine kinase 1 (sTK1) in the treatment of patients withacute exacerbation of chronic obstructive pulmonarydisease (AECOPD). Methods The retrospective treating studies were performed on 104 patients with AECOPD , sTK1 level was de-tected by chemiluminescence dot-blot assay before and after treatment , and the correlation between the expression of sTK1 and CRP after treatment in the two groups was analyzed. Results The levels of sTK1 and CRP in the ef-fective group significantly lowered after treatment (P < 0.05). The expression of sTK1 in the effective group and in the ineffective group after treatment was significantlycorrelated with CRP (P < 0.05). Conclusion sTK1 can be used as a valuable indicator to evaluate the severity of AECOPD and to estimate the efficacy of AECOPD thera-py.

Objective To investigate the expression of adaptin-2(AP-2) in different stages of mouse coch‐lea and its probable role in the auditory generation and formation .Methods Mice were divided into 4 experimental groups by age (7 days old ,15 days old ,35 days old ,16 months old) ,which respectively represented the newborn mice ,developmental mice ,mature mice and old mice .Auditory brainstem response (ABR) ,laser scanning confocal microscopy (LSCM) ,immune-fluroscence histochemistry and qRT - PCR were used in this study .Results For the 15 days old ,35 days old ,16 months old groups ,ABR average threshold was 18 .67 ± 1 .21 dB nHL ,13 .83 ± 1 .47 dB nHL ,37 .83 ± 7 .68 dB nHL ,respectively ,for the 7 days old groups no responses were observed .The AP-2 im‐munoreactivity was found in all the stages of mice cochlea inner hair cell (IHC) cytoplasm ,especially in IHC basal part ,nearby the ribbon synapse .For the 7 days old ,15 days old ,35 days old ,16 months old groups ,immune-flu‐roscence histochemistry IMV(intensity mean value)were 190 .91 ± 17 .27 ,494 .06 ± 27 .63 ,838 .41 ± 38 .23 ,682 .65 ± 72 .22 ,respectively .For the 7 days old ,15 days old ,35 days old ,16 months old groups ,mRNA RQ(relative quantity)were 0 .53 ± 0 .09 ,1 .03 ± 0 .02 ,1 .00 ± 0 .09 ,1 .03 ± 0 .06 ,respectively .Developmental mice expressed significantly higher than those of the newborn in the AP -2 protein expression level which was measured by immuno -fluores‐cence histochemistry and qRT PCR(P<0 .01) .There was no significant difference between old and mature mice in the AP-2 protein expression level measured by qRT PCR (P> 0 .05) ,but mature mice had significant advantages by immuno-fluorescence histochemistry .Conclusion AP-2 protein expression level may closely related to the au‐ditory formation and maintenance because its expression gradually increases with age in mice .

BACKGROUND: Limb negative pressure treatment can dilate limb vessels and improve terminal microcirculation. P-substance has strong vasodilative activity and is involved in the sensation of the skin to traumatic stimulation and the modulation of local vascular function.OBJECTIVE: To observe the influence of limb negative pressure on cutaneous P-substance immunoreactive nerve fibers in dogs with peripheral arterial occlusive disease.DESIGN: Randomized controlled experiment.SETTING: The 3rd Department of General Surgery, Xijing Hospital Affiliated to the Fourth Military Medical University of Chinese PLA.MATERIALS: This experiment was conducted at the Animal Laboratory of Xijing Hospital Affiliated to the Fourth Military Medical University of Chinese PLA between April 2003 and May 2004. Totally 17 healthy hybrid dogs were randomized into 3 groups, namely, treatment group of 10 dogs,non-treatment group of 5 dogs, and normal control group of 2 dogs.INTERVENTIONS: Negative pressure treatment on affected limbs: After superficial anesthesia, the left hindlimbs of the animals were put into the home-made negative cabin for negative pressure treatment with pressure designed as -12kPa, for 15 minutes, once a day for consecutive 10 days.[1] Treatment group: The left hindlimb ischemic model was prepared 14days before starting 10-day negative pressure treatment; after that the animals were subjected to infusion, the skin of the 2nd toe of affected limbs, as well as L1-5 spinal cords and dorsal root ganglion were obtained for immunohistochemical (IHC) staining. Meanwhile prostaglandin E1 immunoreactive nerve fibers were detected. [2] Non-treatment group: The animals received the same treatment and examination as treatment group except for negative pressure. [3] Normal control group: No ischemic model was prepared or negative pressure treatment was given except for IHC staining.MAIN OUTCOME MEASURES: Changes of cutaneous P-substance immunoreactive nerve fiber in each group. RESULTS: Totally 17 dogs entered the result analysis. Changes of cutaneous P-substance immunoreactive nerve fibers: The cutaneous P-substance immunoreactive nerve fibers in dermis connective tissues and layer vessels were reduced in treatment group compared to those in non-treatment group[(24.70±4.6), (43.49±6.3) μm/mm2, P ＜ 0.01], but still higher than those in control group [(18.10±5.4) μm/mm2, P ＜ 0.01]; dermis P-substance immunoreactive nerve fibers in non-treatment group were more than those in normal control group (P ＜ 0.01), with P-substance immunoreactive nerve fibers increased and deeply stained in dermis connective tissues and small vessels. In contrast, P-substance immunoreactive nerve fibers were not observed in the horny layer but in the dermis of the toe in normal control group.CONCLUSION: Results of the present study suggest that limb negative pressure can enhance P-substance release from cutaneous sensory nerve fibers.

BACKGROUND: Harmful stimuli induce increased production of calcitonin gene-related peptide(CGRP) in the spinal cord and dorsal root ganglion, causing also intense dilation of the microvessels. But it remains unknown whether vessel dilation and pain relief were accompanied by increased CGRP production when negative pressure is applied on the limbs for treatment of peripheral arterial occlusion diseases (PAOD).OBJECTIVE: To examine the changes of calcitonin gene-related peptide (GGRP) -immunoractive nerve fibers in the spinal cord and dorsal root ganglion in dogs with PAOD treated with negative pressure on the limbs.DESIGN: A randomized controlled retrospective study.SETTING: The department of general surgery of a military medical university.MATERIALS: This study was carried out at Xijing Hospital of the Fourth Military Medical University of Chinese PLA between January and August 2003. Seventeen adult male dogs weighing 12 - 18 kg, regardless of the gender, were selected.INTERVENTIONS: Seventeen dogs were randomly divided into three groups, namely the treatment group( n = 10), model group( n = 5), and the normal control group( n = 2). Posterior left leg ischemia was induced in dogs in the treatment and model groups, and those in the treatment group, but not the model group, were treated with negative limb pressure for 10 days 14days after model establishment. The spinal cord and dorsal ganglion at L1-5of these two groups were collected and stained immunohistochemically for observing the changes of GGRP-immunreactive nerve fibers. The dogs in the normal control group were also sampled in similar manner for immunohistochemical staining.MAIN OUTCOME MEASURES: Distribution of CGRP-immunoreactive nerve fibers in the spinal cord and dorsal ganglions of the three groups of dogs.RESULTS: In the dogs of the model group, GGRP-immunoreactive nerve fibers in the spinal cord and dorsal ganglions was significantly more numerous[ (75. 00 ±4. 30)%, (68.20 ± 2.60)% ] than those in the treatment and normal control groups[ (58. 20 ±5. 10)%, (52. 20 ±6.20)%; (37.00±4. 20)%, (34. 00 ± 1.40)%, P ＜ 0.01]. The positive nerve fibers were less strongly stained in the treatment group than those in the model group,but still stronger stained those in the normal control group, with significant difference between the three groups( P ＜ 0.01).CONCLUSION: Negative pressure on the limbs may attenuate the synthesis of CGRP-immunoreactive nerve fibers in the spinal cord and dorsal root ganglion and pain conduction following PAOD in dogs, so that harmful afferent stimuli are inhibited to relieve the pain in the limbs.