Through the map analysis of various metabolites in rat's urine, the biomarkers in syndrome suitable for Small Qing-Long Decoction (SQLD) were found out, which provided objective basis for the clinical diagnosis and treatment. Based on the rat's urine nuclear magnetic resonance (NMR) spectrum detection of the blank control group, the model group and the SQLD group, the data come from the map by MestRenova pretreatment and integration were analyzed using SIMCA-P+ by (PLS-DA). Then, the corresponding scatter diagram and diagram of loading were re-ceived and the differences of metabolites were found out by the comparison . The results showed that compared with the blank control group, contents of creatinine, guanidinoacetic acid, trimethylamine, taurine, allantoin metabolites in the model group had elevated. And the contents of succinic acid, 2-OG, lactic acid, glycine, methyl nicotine metabolite had decreased. After medication, contents of creatinine, trimethylamine, allantoin, taurine of SQLD group were still higher than the blank control group. However, the contents were reduced compared with the model group. The content of lactic acid was lower than the blank control group, but higher than the model group. There was no statistical significance on changes of guanidinoacetic acid, 2-OG, N-nornicotine. It was concluded that substances found were mainly related to energy metabolism, inflammation and immune resistance, which was especially related to energy metabolism change obviously.

Alveolitis, Extrinsic Allergic ; diagnosis ; therapy ; Humans ; Occupational Diseases ; diagnosis ; therapy

Neural cell adhesion molecule (NCAM) is a glycoprotein expressing on the surface of neurons, glial cells, bone cells and natural killer cells. NCAM plays an important role in the process of cell - cell adhesion and cell migration, and is also a model protein to study polysialic acid. In this paper, NCAM gene from mouse mammary gland cells (NMuMG) was cloned into eukaryotic expression vectors pcDNA3.1(+) and transfected into mutant Chinese hamster ovary cells ldlD-14. The stable transfection over-expressing NCAM was obtained through the G418 selection and confirmed by Western blotting. Due to unique characters of ldlD-14 cells, carbohydrate chain of NCAM molecule can be easily manipulated with or without adding galactose in the serum free medium, and this modification can provide the basis for further studies on the effect of glycosylation on NCAM molecular function.
Animals ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Female ; Galactose ; Glycosylation ; Mammary Glands, Animal ; cytology ; Mice ; Neural Cell Adhesion Molecules ; biosynthesis ; Polysaccharides ; chemistry ; Sialic Acids ; chemistry ; Transfection

Objective To observe the protective effect of magnesium gluconate on myocardial apoptosis by ischemia reperfusion injury in isolated rat hearts, and study the possible mechanism. Methods The hearts of 48 Sprague-Dawely rats were isolated, linked to Lange-ndorff perfusion apparatus, and randomly divided into 3 equal groups(n = 16 each) : Control group, ischemia/reperfusion (I/R) group and magnesium gheonate group. 8 rats in each group were perfused. Control group was pedused with modified KH buffer for 110min. I/B group was perfuesd with modified KH buffer for 20 min, then exposed to iscbemia for 30 min, and then reperfused with modified KH buffer for 60 min. Magnesium gheonate group was perfumed with modified KH buffer with magnesium gluconate for 20 min, then exposed to isohemia for 30 min and then reperfused with modified KH buffer with magnesium glueonate for 60 min. Lacate dehydrogenase (LDH) and ereatine kinase (CK) in the effluent liquid from the heart were measured after reperfusion. The concentration of Ca2+ and NO in the left ventricle were determined. The other 8 rats in each group were reperfused for 120 minutes as the method described before. After repeffusion, the myoeyte apoptosis was examined by Annexin-V-FITC/PI. After the two experiments the incidence of ventrieular arrhytlunias during reperfusion was assessed. Results Compared with I/R, magnesium glueonate decreased the incidence of ventricular an'hythmias(P <0. 01). The contents of CK and LDH in the effluent liquid from the heart in magnesium glueonate group was lower than that of I/R group (P <0. 01). The contents of Ca2+ and NO in the left ventricle in magnesium gluconate group was decreased than that of I/R group (P <0. 01). The index of myocyte apoptosis were significanfly lower in magnesium glueonate group than that of I/R group (apoptosis index :27.79±1.59 vs 33.61±2.10, P < 0. 01) . Conclusion Magnesium glueonate has protective effect on myocardial isohemia reperfusion injury in rats. The protective effect may be related to decreasing myocyte apoptosis by increasing the content of NO and relieving calcium overload.

Animals ; Diazepam ; pharmacology ; Drug Antagonism ; Electroencephalography ; Female ; Male ; Pyrazoles ; poisoning ; toxicity ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the changes in the expression of acetylated histone H3 (Ac-H3) and deacetylase silent information regulator 1 (SIRT1) in dorsal root ganglions in a rat model of negative phenotype neuropathic pain.Methods Forty-eight male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 2 groups (n =24 each):sham operation group (group S) and C-fiber dysfunction group (group CFD).The rats were anesthetized with 10％ chloral hydrate 3 ml/kg.C-fiber dysfunction was induced by exposing sciatic nerve to 8％ capsaicin for 30 min in group CFD.The thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) were measured before and on 1,3,7 and 14 days after CFD.Six rats were then sacrificed at each time and the lumbar segments (L5) of the dorsal root ganglions were removed for detection of SIRT1 mRNA expression (by RT-PCR) and Ac-H3 and SIRT1 protein expression (by Western blot).Results Compared with group S,TWL was significantly increased at 1,3,7 and 14 days after CFD,SIRT1 mRNA and protein expression was up-regulated and Ac-H3 expression was down-regulated at 3,7 and 14 days after CFD (P ＜ 0.05),while no significant change was found in MWT at each time point in group CFD (P ＞ 0.05).Conclusion The mechanism of negative phenotype neuropathic pain is related to up-regulation of deacetylase SIRT1 expression and decreased acetylation of histone H3 in rat dorsal root ganglions.

Objective To observe the effects of the compatibility of Radix Astragali and Radix Angelicae Sinensis in different proportion on hemodynamic and balance of Th1/Th2 cells of Qi deficiency and blood stasis rat model. Methods Qi deficiency and blood stasis rats were caused by restricted diet, forced swimming and norepinephrine subcutaneous injecting, and treated by compatibility of Radix Astragali and Radix Angelicae Sinensis in different proportion of 5∶1, 3∶1, 1∶1 and 1∶2 for 21 days. The indexes of hemorheology were detected with hemorheological analyser, and the level of IFN-γand IL-4 in serum were tested by ELISA. Results Compared with model group, the compatibility of Radix Astragali and Radix Angelicae Sinensis in proportion of 1∶1, 3∶1 and 5∶1 groups reduced the low shear blood viscosity. The spleen index of model group decreased, and compatibility of Radix Astragali and Radix Angelicae Sinensis in 5∶1 proportion group increased spleen index. The level of IL-4 increased and IFN-γ decreased in the serum of model group, the compatibility of Radix Astragali and Radix Angelicae Sinensis in proportion of 3∶1 group increased the level of IFN-γ. Astragalus angelica 5∶1 group decreased the level of IL-4 and increased the level of IFN-γ. Conclusion The compatibility of Radix Astragali and Radix Angelicae Sinensis can improve hemorheology, adjust the balance of Th1/Th2 cells of Qi deficiency and blood stasis rats. The effects were better when Radix Astragali’s dosage greater than that of Radix Angelicae Sinensis, and the group of 5∶1 proportion was the best.

Objective To investigate the mechanism of regulating gastrointestinal motility by elec- troacupunturing point of"Zusanli"in rabbits.Methods Thirty adult rabbits were divided into control, electrical acupuncture point of"Zusanli"and non acupuncture point groups.The blood concentrations of motilin were detected at different times(before acupuncture and 15,30,45 and 60 min after acupunc- ture).Sixty min after acupuncture,the mice were sacrificed.The acetylcholine esterase and nitric oxide synthase(NOS)in gastric and jejunum tissues were examined.The electron microscope was used to observe the vesicles of nerve ending.Results The concentrations of motilin in electrical acupuncture group was significantly increased and reached the high peak at 45min.The activities of acetylcholine es- terase was increased significantly in electrical acupuncture group compared to control group[gastric tis- sue:(15 571?2876)pinel vs(9081?801)pinel,P

Objective To observe hemodynamic characteristics and the correlation with syndrome types of traditional Chinese medicine (TCM) in patients with septic shock, so as to direct the treatment based on syndrome differentiation. Methods A prospective observation was conducted. Sixty-eight patients with septic shock admitted to the Department of Critical Care Medicine of Dade Road General Hospital of Guangdong Hospital of TCM from January 2013 to July 2015 were enrolled. Pulse indicating continuous cardiac output (PiCCO) was used to monitor the hemodynamic changes, including heart rate (HR), mean arterial pressure (MAP), central venous pressure (CVP), cardiac index (CI), global end diastolic volume index (GEDVI), extravascular lung water index (EVLWI), maximum rate of the increase in pressure (dPmax) and systemic vascular resistance index (SVRI), for assessment of hemodynamics. According to the CI, the patients were divided into two groups , i.e. high CI group (CI ≥ 50.0 mL·s-1·m-2, n = 34) and low CI group (CI < 50.0 mL·s-1·m-2, n = 34), and the clinical and hemodynamic characteristics of two groups were investigated. The TCM differentiation was conducted with four syndromes and four methods, and the hemodynamic characteristics of different syndrome types were investigated, the correlation between the TCM syndrome factors and hemodynamic parameters was analyzed. The patients were divided into survival group and death group, and clinical parameters and hemodynamic characteristics were compared between two groups. Results The acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score and blood glucose of low CI group were higher than those of high CI group [APACHEⅡ score: 24.4±7.2 vs. 19.8±7.4, t = -2.279, P = 0.023; blood glucose (mmol/L): 9.7 (7.7, 14.6) vs. 6.7 (5.6, 10.0), Z = -2.257, P = 0.024], CI and GEDVI were lowered [CI (mL·s-1·m-2): 36.7±8.3 vs. 68.4±16.7, t = 10.285, P = 0.000; GEDVI (mL/m2): 689.0 (566.0, 883.8) vs. 838.5 (692.8, 1 247.3), Z = -2.711, P = 0.007], while SVRI was increased [kPa·s·L-1·m-2: 248.7 (202.1, 324.5) vs. 143.4 (102.7, 171.4), Z = -5.336, P = 0.000]. Accompanied symptoms were found to occur more commonly in septic shock patients, and the most common syndrome elements were Qi deficiency syndrome (n = 45), blood stasis syndrome (n = 40), heat-toxin syndrome (n = 37), Fushi syndrome (n = 24) and Yin deficiency syndrome (n = 10), respectively. There was no significant difference in hemodynamic parameters among patients with five types of syndrome (all P > 0.05). However, only the CI of those with Qi deficiency syndrome was significantly lower than that of heat-toxin syndrome (mL·s-1·m-2: 48.3±18.3 vs. 53.3±21.7, P < 0.05). While the results after removing the effect of accompanied symptoms showed that CI of Qi deficiency syndrome was significantly lower than that of non-Qi deficiency syndrome (mL·s-1·m-2: 48.3±18.3 vs. 61.7±21.7, t = -2.783, P = 0.007), CI of heat-toxin syndrome was significantly higher than that of non-heat-toxin syndrome (mL·s-1·m-2: 58.3±21.7 vs. 48.3±16.7, t = 2.133, P = 0.037), EVLWI of blood stasis syndrome was significantly lower than that of non-blood stasis syndrome [mL/kg: 10.0 (7.0, 15.1) vs. 14.9 (8.5, 26.8), Z = -2.075, P = 0.038]. Compared with survival group (n = 38), APACHEⅡ score in death group (n = 30) was increased (25.8±8.4 vs. 19.1±5.4, t = -3.940, P = 0.000), the proportion of continuous renal replacement therapy (CRRT) was increased [60.0% (18/30) vs. 31.6% (12/38), χ2 = 5.493, P = 0.019], HR was increased (bpm: 118.5±20.5 vs. 98.1±19.9, t = -4.157, P = 0.000), and the proportion of Qi deficiency syndrome was increased [86.7% (26/30) vs. 50.0% (19/38), χ2 = 10.070, P = 0.002]. Conclusions Patients with sepsis shock may be divided into high-output and low-resistance & low-output and high-resistance groups according to hemodynamics, with respective hemodynamic characteristics. Hemodynamic performance differed among different syndrome types, and there was a certain relationship. Hemodynamic monitoring with PiCCO was a useful supplement of TCM, which was good for the evidence-based medicine.

Objective Abnormal activation of mitogen-and stress-activated kinase (MSK1) plays an important role in the development of various cancers.This study was to explore the effect of small interfering RNA (siRNA)-mediated MSK1-silencing on the proliferation of human nasopharyngeal carcinoma (NPC) cells and its underlying mechanism.Methods The siRNA vector targeting MSK1 was constructed and transfected into CNE2 cells, and the NPC cell line stably expressing MSK1 was established.Then the cells were divided into a blank control (without transfection of the plasmid), a negative control (with stable transfection of the negative control plasmid), and an experimental group (with stable transfection of the positive recombinant plasmid).The expressions of MSK1 mRNA and protein were detected by real-time quantitative PCR and Western blot, respectively, the proliferation of the cells determined by CCK-8 and colony formation assays, the cell cycles analyzed by flow cytometry, the level of histone H3 phosphorylation at Ser10 examined by Western blot, and The transcriptional activity and expression of the c-jun protein measured by dual-luciferase reporter gene assay and Western blot.Results Compared with the blank control, the inhibition rates of cell proliferation at 48, 72 and 96 hours were significantly reduced in the experimental group (P<0.05), and so were the colony formation ability of the cells (P<0.01) and the expression and transcriptional activity of the c-jun protein (P<0.05).In comparison with the negative control, the experimental group showed significant decreases in the rate of cell growth after 24 hours, the inhibition rates of cell proliferation at 48, 72 and 96 hours (P<0.05), the number of formed colonies ([221.00±20.08] vs [99.67±15.57] / 300 cells, P<0.01), the proportion of S-phase cells (P<0.01), and the expression of the c-jun protein in the CNE2 cells ([100.00±0.00] vs [48.77±10.71] %, P<0.05), but a remarkable increase in the percentage of G0/G1-phase cells (P<0.01).Furthermore, histone H3 phosphorylation at Ser10 was markedly reduced (P<0.01) but no significant change was observed in the expression of the total c-jun protein in the experimental group.Conclusion Knockdown of MSK1 using siRNA can significantly inhibit the growth and proliferation of CNE2 cells, which may be closely related to the decreased phosphorylation of histone H3 and subsequently down-regulated transcriptional activity of c-jun.