OBJECTIVEEffects of ginsenoside Rb1, Rg1 and Re on neurotrophic factor signal transduction pathway using liposome-mediated transfection of eukaryotic cells approach.

METHODThe injury model was established by treating SH-SY5Y cells with 0.6 mmol x L(-1) of corticosterone (CORT) by 24 h. SH-SY5Y cell were pretreated with CORT for 30 min followed by co-treated with 120,60 and 20 micromol x L(-1) of Rb1, 120, 80 and 40 micromol x L(-1) of Rg1 and 120, 80 and 40 micromol x L(-1) of Re for 24 h. Cells viability was determined by Cell Counting Kit (CCK) assay. CREB expressing Luciferase reporter gene was constructed and transfected with plasmid containing hRaf, hcAMP, hAkt, hCaMK gene into human embryonic kidney (HEK293) cells using liposornal transfection reagent lipofection 2000. The expression of CREB before and after it addion of Rb1, Rg1 and Re was examined by Luc assay system and Western blotting.

RESULTCompared with normal control group, CORT significantly decreased the viability of SH-SY5Y cells to 67.21% (P < 0.01). CCK results show that Rb1 (60 micromol x L(-1)), Rg1 (80 micromol x L(-1)) and Re (80 micromol x L(-1)) on SH-SY5Y cells have significant protective effect (P < 0.01). Lucassay and Western blotting results show that the gene and protein levels of CREB increased significantly through the pathway of Raf and Akt with Rb1 and Rg1 (P < 0.01), Re can increase significantly the gene and protein levels of CREB through the pathway of Raf and CaMK II.

CONCLUSIONRb1, Rg1 and Re protects SH-SY5Y cells from CORT-induced damage and the neuroprotective mechanism may be associated with the Raf-CREB, Akt-CREB and CaMK II -CREB pathways.

Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; genetics ; metabolism ; Cell Line ; Cell Survival ; drug effects ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Genes, Reporter ; Ginsenosides ; pharmacology ; Humans ; Panax ; chemistry ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Signal Transduction ; drug effects ; raf Kinases ; genetics ; metabolism

Dingzhi Xiaowan is a widely used traditional Chinese medicine in treating depression, which is a similar formula of Kaixinsan. In this research, a rapid ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS(E)) method was established to analyze the metabolites of Dingzhi Xiaowan in depressive model rat plasma, bile, urine and feces. After we established Chronic unpredictable mild stress (CUMS) model rats and orally administrated Dingzhi Xiaowan, rat plasma, bile, urine and feces samples were collected and prepared. Using Waters Cortects UPLC C18 column (2.1 mm x 50 mm, 1.6 μm), acetonitrile-0.1% formic acid mobile phase gradient, these samples were analyzed and 33 metabolites of nine bioactive compounds were detected and tentatively identified by Metabolynx. Among the 33 metabolites, three metabolites were identified from plasma sample, three came from bile sample, and 27 metabolites were identified from urine and feces samples. This approach provided a rapid method for characterizing the metabolites of Dingzhi Xiaowan and gave the truly active structures and the action mechanism of their antidepressant effects.
Animals ; Bile ; metabolism ; Chromatography, High Pressure Liquid ; Depression ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; metabolism ; Feces ; chemistry ; Male ; Mass Spectrometry ; Medicine, Chinese Traditional ; Plant Extracts ; metabolism ; Rats ; Rats, Sprague-Dawley

The efficacy of Chinese herbal formula in treating depression has been proved in many studies. In this study, six different Kaixin San formulas were compared to investigate their effects on central monoamine neurotransmitters of chronic stress rats and against depression based on their different components in plasma, in order to discuss the efficacy-comparability relationship and the possible efficacy mechanism. The classic isolation method and the chronic unpredictable mild stress (CUMS) depression model were combined to investigate the changes in contents in hippocampus and monoamine neurotransmitters (NE, DA, 5-HT) and the components of some formulas in plasma with HPLC and UPLC-Q-TOF-MSE methods. As a result, Dingzhi Xiaowan recorded in Essential Recipes for Emergent Use Worth A Thousand significantly increased the behavioral scores, NE and 5-HT contents in hippocampus and NE, DA and 5-HT contents in cortex, with the best anti-depressant effect. Dingzhi Xiaowan recorded in Complete Records of Ancient and Modern Medical Works showed a notable increase in sucrose preference and open field score in model rats, NE content in hippocampus and NE, DA and 5-HT contents in cortex, with a certain anti anti-depressant effect. Kaixin San recorded in Ishinpo showed remarkable rise in weight of model rats. NE content in hippocampus and DA content in cortex. Puxin Decoction recorded in A Supplement to Recipes Worth A Thousand Gold showed 5-HT content in hippocampus and DA content in cortex. Kaixin San recorded in Yimenfang only showed DA content in cortex. Kaixin Wan recorded in Essential Recipes for Emergent Use Worth A Thousand did not mention the antidepressant effect. According to the results, the formulas' different anti-depressant effects may be related to the different plasma components.
Animals ; Behavior, Animal ; drug effects ; Biogenic Monoamines ; analysis ; Brain Chemistry ; drug effects ; Chronic Disease ; Drugs, Chinese Herbal ; pharmacology ; Male ; Medicine, Chinese Traditional ; Neurotransmitter Agents ; analysis ; Norepinephrine ; analysis ; Rats ; Rats, Sprague-Dawley ; Serotonin ; analysis ; Stress, Psychological ; metabolism

Objective:To analyze the relationship between free androgen index and insulin function in obese young men aged from 20 to 35.Methods:A total of 82 young obese men in Obesity Clinic from February to October 2019 were enrolled in the study. The subjects were divided into 3 subgroups according to free androgen index level tertiles. The blood glucose and insulin levels were tested after oral glucose tolerance test. Homeostasis model assessment for insulin resistance (HOMA-IR), homeostasis model assessment for β cell function (HOMA-β), insulin secretion index, and insulin sensitivity index (Matsuda index) were used to evaluate β cell function in oder to analyze the relationship between free androgen index and insulin function.Results:In young obese men, participants with higher free androgen index levels exhibited less waist circumference, lower body mass index, 1 h postprandial insulin, 2 h postprandial insulin, HOMA-IR level but with a higher total testosterone, sex hormone binding globulin, and Matsuda index level (all P<0.05). There was a negative correlation between the free androgen index and the HOMA-IR ( r=-0.386, P=0.016), and the correlation tended to a linear trend after adjustment for age, sex, body mass index, and waist circumference ( Ptrend=0.034). Free testosterone was positively correlated with Matsuda index ( r=0.280, P=0.004), but the correlation disappeared after adjustment ( Ptrend=0.623). The results of further regression analysis showed that the level of free testosterone index decreased by 14.1% ( OR=0.869, 95% CI0.767-0.984, P=0.028) for each increase of HOMA-IR after adjustment. Conclusion:The level of free testosterone index is a predictor of insulin resistance in obese young men, but the association between this parameter and insulin sensitivity may be caused by obesity.

Objective To determine if Shenfu injectio(SFI)has any analeptic action in patients emerging from general anesthesia.Methods Eighty-six ASAⅠorⅡadult patients undergoing elective abdominal surgery under general anesthesia were randomly divided into 2 groups(n=43 each):SFI group and control group.The patients were premedicated with intramuscular phenobarbital 0.1g and atropine 0.5mg.Anesthesia was induced with propofol 2 mg?kg~(-1),fentanyl 4-5?g?kg~(-1) and vecuronium 0.1 mg?kg~(-1) and maintained with propofol infusion(2-4 mg?kg~(-1)?h~(-1)),0.5%-1.0% isoflurane inhalation and intermittent i.v.boluses of fentanyl and vecuronium.The patients were intubated and mechanically ventilated.The propofol infusion and isoflurane inhalation were stopped during skin closure.The patients were still unconscious and on mechanical ventilation at the end of surgery and transferred to PACU with a tube in trachea.As soon as the patients reached the PACU,SFI 1 ml?kg~(-1) in Ringer's solution 100 ml was infused over 10 min.In control group the patients received Ringer's solution 100 ml without SFI.The following times were recorded:(1)the time when the patients opened their eyes on command;(2)the time when mechanical ventilation was stopped;(3)the time when oxygen inhalation was stopped;(4)the extubation time;(5)the time of staying in PACU.Venous blood samples were taken before(T_0) and 5,15 and 45 min(T_(1,2,3))after SFI infusion for determination of plasma?-endorphin concentration.Results The awakening time,the mechanical ventilation time,oxygen inhalation time,extubation time and duration of PACU stay were significantly shorter in SFI group than in control group.There were no significant differences in MAP and HR after SFI between the two groups.The plasma?-endorphin concentration was significantly higher in group SFI than in control group.Conclusion Shenfu injectio can make patients emerging from general anesthesia faster.

OBJECTIVETo determine the visible light-induced photodegradation kinetics of two xanthene photosensitizers, phloxine B and uranine, in solution and on the surface of silica TLC plates, and to examine the phototoxicity of residues of degradation, which could provide valuable safety data on the two photosensitizers and other xanthene chemicals when applied in the environment.

METHODSUV-Vis absorption during photodegradation was monitored with a Unico 2102 spectrophotometer. Organic content of samples was measured with a Shimadzu TOC 4100. Phototoxicity tests were carried out using Saccharomyces cerevisiae with the methods modified from Daniels.

RESULTSWhen phloxine B and uranine degraded in solution, their apparent rate constant k was 0.0019 and 0.0027 min(-1), respectively. The total organic carbon (TOC) content decreased by approximately 50% during the 8 h irradiation period, which led to a gradual decrease in phototoxicity of the residues. The photodegradation of photosensitizers on the surface of silica TLC plates was much faster than that in the solution. The apparent rate constant k and the half life of phloxine B were 0.0073 min(-1) and 95 min, respectively.

CONCLUSIONVisible light can rapidly induce photodegradation of phloxine B and uranine. The phototoxicity of residues is also decreased. The environmental risk of applications of phloxine B and uranine is minimal.

Eosine I Bluish ; chemistry ; toxicity ; Fluorescein ; chemistry ; toxicity ; Kinetics ; Molecular Structure ; Photolysis ; Photosensitizing Agents ; toxicity ; Saccharomyces cerevisiae ; drug effects ; radiation effects

Objective To determine the visible light-induced photodegradation kinetics of two xanthene photosensitizers, phloxine B and uranine, in solution and on the surface of silica TLC plates, and to examine the phototoxicity of residues of degradation, which could provide valuable safety data on the two photosensitizers and other xanthene chemicals when applied in the environment. Methods UV-Vis absorption during hotodegradation was monitored with a Unico 2102 spectrophotometer. Organic content of samples was measured with a Shimadzu TOC 4100. Phototoxicity tests were carded out using Saccharomyces cerevisiae with the methods modified from Daniels. Results When phloxine B and uranine degraded in solution, their apparent rate constant k was 0.0019 and 0.0027 min-1, respectively. The total organic carbon (TOC) content decreased by approximately 50% during the 8 h irradiation period, which led to a gradual decrease in phototoxicity of the residues. The photodegradation of photosensitizers on the surface of silica TLC plates was much faster than that in the solution. The apparent rate constant k and the half life of phloxine B were 0.0073 min-1 and 95 min, respectively. Conclusion Visible light can rapidly induce photodegradation of phloxine B and ranine. The phototoxicity of residues is also decreased. The environmental risk of applications of phloxine B and uranine is minimal.

Objective To purify Shiga toxin Ⅱ (STX Ⅱ) of enterohaemorrhagic Escherichia coli (EHEC) O157: H7 by affinity chromatography, and characterize its biological function. Methods The immno-affinity chromatography column was prepared by STX Ⅱ A subunit-specific antibody S1D8 coupling to Sepharose 4B matrix. The purity and specificity of STX Ⅱ molecule secreted by EHEC O157:H7 were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, respectively. The purified toxin was serially diluted and the toxic activities to Vero cell line and mice were observed. The 50% cytotoxic dose (CD50) for Vero cell line and 100% lethal dose (LD100) for mice were calculated. The protection effect of anti-STX Ⅱ polysera to the mice against the purified toxin challenge was also observed. Results STX Ⅱ was successfully purified from culture supernatant of EHEC O157:H7 using affinity chromatography scheme. The relative molecular weights of STX Ⅱ A and B subunits were 32 000 and 7 500 confirmed by SDS-PAGE, respectively. The purified toxin could react with monoclonal antibodies against STX Ⅱ A and B subunits, respectively.The toxin was cytotoxic to Vero cell with CD50 of 20 ng/L and lethal to mice with LD100 of 5 ng.The toxin could be neutralized by anti-STX Ⅱ polysera in vivo. Conclusion STX Ⅱ is successfully purified and its toxic effects are confirmed in both cell line and mouse model.

To simplify the macrocyclic fragment and to modify the zinc binding group of the natural product apicidin, two series of S-hexyl (heptyl) ethanethioate derivatives were designed and synthesized. Twenty-six compounds were synthesized and confirmed with 1H NMR, IR, MS and HR-MS spectrum, which were not reported. Take vorinostat as control, their antiporliferative activities against cancer cell lines, MCF-7 and HL-60, were tested with MTT assay or trypan blue staining method. Generally in both series it was found that, the chiral carbon atom at 7 position is not necessary, compounds II-1, II-3, II-6 and II-13 showed good activity on HL-60 cells in vitro, with the IC50 values less than 10 micromol x L(-1). II-7 and II-8 showed stronger activity against MCF-7 than Vorinostat, with the IC50 of 3.19 and 6.29 micromol x L(-1), respectively.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Proliferation ; drug effects ; Drug Screening Assays, Antitumor ; HL-60 Cells ; Histone Deacetylase Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; Humans ; Inhibitory Concentration 50 ; MCF-7 Cells ; Peptides, Cyclic ; chemical synthesis ; chemistry ; pharmacology ; Structure-Activity Relationship

Adenoids ; diagnostic imaging ; pathology ; Child ; Child, Preschool ; Female ; Humans ; Hypertrophy ; Magnetic Resonance Imaging ; Male ; Radiographic Image Enhancement ; methods ; Tomography, Spiral Computed ; methods