Objective:To determine the equilibrium solubility and the apparent oil/water partition coefficient of nebivolol hydrochloride to provide experimental basis for the development of new preparations.Methods:The concentration of nebivolol hydrochloride was determined by an HPLC method,and a saturated solution method and a shake-flask method were respectively applied to determine the equilibrium solubility and the apparent oil/water partition coefficient of nebivolol hydrochloride in water,0.1 mol·L-1 HCl solution and phosphate buffer solution with different pH values(pH2.0,pH6.8,pH7.4 and pH8.0).Results:At (37±0.5)℃,the equilibrium solubility of nebivolol hydrochloride in water and in 0.1 mol·L-1 HCl solution was 722.53 μg·ml-1and 56.07μg·ml-1,respectively.The apparent oil/water partition coefficient (Log P) of nebivolol hydrochloride was 1.17 and 1.32,respectively.Within the pH range of 2.0-7.4,with the increase of pH value, the equilibrium solubility and the Log P decreased and increased,respectively,while pH value increased from 7.4 to 8.0,the equilibrium solubility of nebivolol hydrochloride increased and Log P decreased.Conclusion:The method is accurate and reliable.Nebivolol hydrochloride has poor water solubility,and the equilibrium solubility and the Log P are both influenced by pH values.

Objective: To observe the clinical efficacy of moxibustion therapy plus Liu's pediatric massage (tuina) for children with recurrent respiratory tract infections due to qi deficiency of spleen and lung. Methods: A total of 60 children who met the inclusion criteria were divided into an observation group and a control group according to the visiting sequence, with 30 cases in each group. Children in the observation group were treated with moxibustion therapy plus Liu's pediatric massage, and those in the control group were treated with Liu's pediatric massage alone. The incidence of respiratory tract infections and traditional Chinese medicine (TCM) symptoms score were observed and recorded in both groups before and after treatment. And the clinical efficacy was compared between the two groups. Results: The total effective rate of the observation group was 93.3％, and that of the control group was 83.3％. The difference between the two groups was statistically significant (P<0.05). After treatment, the TCM symptoms score and total times of infections in both groups were all statistically different from those before treatment (all P<0.05). The differences in TCM symptoms score and infection frequency before and after treatment in the observation group were statistically different from those in the control group (both P<0.05). Conclusion: Moxibustion therapy plus Liu's pediatric massage has a better effect in improving the clinical symptoms and reducing the frequency of respiratory tract infections for children with recurrent respiratory tract infections due to qi deficiency of spleen and lung than the pediatric massage alone.

OBJECTIVETo explore the mechanism of anti-proliferative effect of indomethacin (IN) on chronic myelogenous leukemia (CML) cells.

METHODSMTT was applied to assay CML cells viability under IN intervention. STAT1, STAT5 proteins were analyzed by Western blot, the expressions of phosphorylated STAT1 or STAT5 by immunoprecipitation combined with Western blot, the cellular localization of p-STATs proteins by indirect immunofluorescence technique, and the detection of Bcl-X(L) and COX-2 protein by Western blot.

RESULTSIN could significantly inhibit the viability of CML cells. 0 approximately 400 micromol/L of IN could down-regulate the expression of p-STAT1 or p-STAT5 in a dose-response manner, p-STATs were distributed mainly in the nucleus as scattering spots. The expression of COX-2 protein could be detected in K562 cells. Both Bcl-X(L) and COX-2 proteins could be inhibited by IN in a dose-dependent manner.

CONCLUSIONSIN could significantly inhibit the proliferation of CML cells, the mechanism of which might be related to the suppression of STATs/Bcl-X(L) signal transduction pathway. There exists COX-2 protein expression in K562 cells, the anti-leukemia effect of IN was possibly dependent on COX-2 pathway.

Blotting, Western ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; metabolism ; Cyclooxygenase Inhibitors ; pharmacology ; Down-Regulation ; drug effects ; Humans ; Indomethacin ; pharmacology ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; Microscopy, Fluorescence ; Phosphorylation ; drug effects ; STAT1 Transcription Factor ; metabolism ; STAT5 Transcription Factor ; metabolism ; bcl-X Protein ; metabolism

This study was aimed to observe whether expressions of JAK2, STAT1, STAT5 proteins in indomethacin-treated CML cells involved in the proliferation inhibition of CML cells, and elucidate the pharmacological mechanism of indomethacin anti-leukemia. MTT assay and trypan blue dye exclusion test were used to detect the inhibitory effect of indomethacin on CML cells proliferation. JAK2, STAT1, STAT5 proteins were analyzed by Western blot; the subcellular distribution of STAT1, STAT5 were detected with indirect immunofluorescence technique. The results showed that indomethacin at >or= 400 micromol/L significantly inhibited the proliferation of CML cells and down-regulated the expression of STAT1, STAT5 protein, no JAK2 change was observed. STAT1 and STAT5 were located in cytoplasm. It is concluded that indomethacin inhibits the proliferation of CML cells and the mechanism may be related to down-regulated expression of STAT, or blockage of cells growth signals.
Antineoplastic Agents ; pharmacology ; Blotting, Western ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Fluorescent Antibody Technique ; Humans ; Indomethacin ; pharmacology ; Janus Kinase 2 ; metabolism ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; STAT1 Transcription Factor ; metabolism ; STAT5 Transcription Factor ; metabolism ; Signal Transduction ; drug effects

Using the concepts and methods of epigenetics and metabolomics, to investigate the overall action molecular mechanism of Chrysanthemi indici C (CIC), the anti-hepatitis B virus (HBV) active extracts from Flos chrysanthemi indici. The inhibitory effects of CIC on proliferation and hepatitis B surface antigen (HBsAg), hepatitis B envelope antigen (HBeAg) and HBV-DNA of HepG2.2.15 cells were detected by CCK-8 and antigen kit. The DNA methyltransferases (DNMTs)/ten-eleven-translocation-2 (TET2) equilibrium was detected by ELISA. Illumina 850K methylation chip, pyrosequencing and qPCR were used to determine the action pathway and target of CIC by GO and KEGG analysis. Cell metabolites were extracted with 80% methanol, and the changes of differential metabolites, differential metabolic pathways and cell microenvironment were detected by LC-MS and other metabolomics methods. The results showed that CIC could inhibit the proliferation, HBsAg, HBeAg and HBV-DNA of HepG2.2.15 cells obviously, down-regulate DNA methyltransferase 1 (DNMT1), DNA methyltransferase 3a (DNMT3a) and DNA methyltransferase 3b (DNMT3b), up-regulate TET2, and restore the balance of DNMTs/TET2. The action targets of CIC were phospholipase C gamma 2 (PLCG2), phosphoinositide-3-kinase regulatory subunit 3 (PIK3R3), 1-acylglycerol-3-phosphate O-acyltransferase 2 (AGPAT2), 5-hydroxytryptamine receptor 2B (HTR2B), nerve growth factor (NGF), mainly involved in lipid metabolism, inflammation mediated regulation of transient receptor potential (TRP), phospholipase D signaling and advanced glycation end product-receptor for AGE (AGE-RAGE) signaling in diabetic complications pathways. CIC could significantly affect fatty acid metabolism and had great influence on phenolic acid, alkaloid and lipid metabolites in cell microenvironment. These results suggest that the action mechanism of CIC may be the synergistic action of multiple pathways and multiple targets, including related inflammatory pathways, immune pathways and lipid metabolism, through regulating epigenetic expression balance and restoring the balance of cell microenvironment.

OBJECTIVETo determine whether homeostatic conditions (pH, glycine or ion concentration) affect the protective effects of edaravone on ischemic injury in rat cortical neurons.

METHODSIn cultured rat cortical neurons, the compositions in the experimental solutions were changed to mimic the disturbance of homeostasis after cerebral ischemia. In vitro ischemic injury was induced by oxygen-glucose deprivation (OGD) for 3 h and reperfusion for 12 h, and the neuron injury was evaluated by 3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyl tetrazolium bromide (MTT) reduction assay and lactate dehydrogenase (LDH) release. Effect of edaravone on OGD injury was observed in different experimental solutions.

RESULTIn weak alkalified solution (pH 7.8) or the solution containing glycine (10 micromol/L), OGD injury became more serious; but in weak acidic (pH 6.5) or higher Mg(2+) (1.8 mmol/L) solutions, OGD injury was attenuated. Edaravone (1 micromol/L) reversed the injury in the solutions with pH 6.1,7.4 and 7.8 or the solution containing glycine, but did not show protective effect in the solution with pH 6.5 and the higher Mg(2+) or lower Ca(2+) solution.

CONCLUSIONThe changes of homeostatic conditions affect the severity of ischemic injury of neurons and the protective effect of edaravone.

Animals ; Animals, Newborn ; Antipyrine ; analogs & derivatives ; pharmacology ; Cell Hypoxia ; Cells, Cultured ; Cerebral Cortex ; cytology ; Glycine ; pharmacology ; Homeostasis ; Hydrogen-Ion Concentration ; Magnesium ; pharmacology ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Rats ; Reperfusion Injury ; prevention & control

OBJECTIVETo explore the methods for labeling CDTPA-coupled CD45 monoclonal antibody (mAb) with yttrium-90 ((90)Y) for potential acute myeloid therapy.

METHODSCD45 mAb was labeled with (90)Y by CDTPA and the labeling rate, radiochemical purity, final specific activity, and immunological activity of the mAb were detected.

RESULTSWith the optimal molar ratio of CDTPA/Ab at 20:1, the labeling rate was 95%, radiochemical purity 99.8%, and final specific activity 1.9 mCi/mg. This conjugate was stable in vitro with comparable immunological activity in comparison with unlabeled CD45 mAb.

CONCLUSION(90)Y-CDTPA-CD45 mAb possesses good properties as an ideal targetting therapeutic agent for acute leukemia.

Anhydrides ; chemistry ; Antibodies, Monoclonal ; chemistry ; immunology ; Humans ; Immunoconjugates ; chemistry ; immunology ; Isotope Labeling ; methods ; Leukocyte Common Antigens ; immunology ; Pentetic Acid ; chemistry ; Radiopharmaceuticals ; chemical synthesis ; chemistry ; immunology ; Yttrium Radioisotopes ; chemistry

OBJECTIVETo assess the antitumor efficacy and adverse effects of bortezomib either used alone or in combination with arsenic trioxide for transplanted tumor in nude mice.

METHODSNude mice bearing HL-60 cell xenografts were randomized into 4 groups to receive treatment with normal saline, bortezomib, arsenic trioxide, bortezomib plus arsenic trioxide. The tumor growth inhibition and general condition of the nude mice were observed, and in situ TUNEL assay and immunohistochemistry were performed on the transplanted tumors.

RESULTSBortezomib alone and in combination with arsenic trioxide could both inhibit the growth of the transplanted tumors, prolong the survival of the nude mice, and induce cell apoptosis and growth inhibition of the HL-60 cells in vivo, and the combined administration exhibited even better effects. The administration was well tolerated with causing manifest vital organ damages in the mice.

CONCLUSIONBortezomib in combination with arsenic trioxide has significant antitumor effect in nude mice bearing HL-60 cell xenografts possibly by inducing HL-60 cell apoptosis and growth inhibition without producing no significant adverse effects.

Animals ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Proliferation ; Disease Models, Animal ; HL-60 Cells ; Humans ; Leukemia ; drug therapy ; physiopathology ; Male ; Mice ; Mice, Nude ; Oxides ; pharmacology ; Pyrazines ; pharmacology ; Random Allocation ; Xenograft Model Antitumor Assays

OBJECTIVETo investigate the effect of bortezomib alone or combined with harringtonine (HT) or arsenic trioxide (As2O3) on the proliferation capacity and apoptosis of HL-60/ADM cell line and fresh cells from refractory/relapse acute leukemia patients.

METHODSHL-60/ADM cells or refractory/relapse leukemia cells were incubated with bortezomib at different doses alone and in combination with HT or As2O3. The proliferation capacity was observed by MTT assay, cell apoptosis by fluorescence microscopy and flow cytometry. Intracellular concentration of daunorubicin (DNR) was determined by flow cytometry.

RESULTSIn bortezomib-treated HL-60/ADM cells, the proliferation inhibition rate and apoptotic cells increased in a time- and dose-dependent manner. 40 nmol/L bortezomib could maximally inhibit the proliferation of HL-60/ADM cells at 48 hours. 15 micromol/L As2O3 or 752 nmol/L HT combined with different doses of bortezomib could inhibit proliferation and induce apoptosis of HL-60/ADM cells. The As2O3 plus bortezomib or HT plus bortezomib showed a greater anticancer efficacy than either of the drugs alone (P < 0.05, P < 0.01). Bortezomib (10 nmol/L) could markedly enhance the intracellular accumulation of DNR in HL-60/ADM cells (P < 0.05).

CONCLUSIONSBortezomib can inhibit proliferation and induce apoptosis of HL-60/ADM cells and fresh refractory/relapse acute leukemia cells, especially combined with HT or As2O3.

Adolescent ; Adult ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Child ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; HL-60 Cells ; Harringtonines ; pharmacology ; Humans ; Male ; Oxides ; pharmacology ; Pyrazines ; pharmacology ; Young Adult

The aim of this study was to investigate the effect of bortezomib alone and in combination with harringtonine on apoptosis of HL-60 cells. HL-60 cells were treated with bortezomib, harringtonine in different concentrations for 12 - 48 hours. Cell proliferation was analyzed by MTT assay; the apoptosis of HL-60 cells was observed by DNA gel electrophoresis, fluorescence microscopy and flow cytometry. The results showed that 10 - 50 nmol/L bortezomib could effectively inhibit HL-60 cell proliferation, and induced its apoptosis. After treating for 12 hours, 10 nmol/L bortezomib could trigger cells apoptosis. With time prolongation or dose increase, HL-60 cell apoptotic rate significantly increased. Furthermore, co-administration of bortezomib (10 nmol/L) with harringtonine (30 nmol/L) resulted in a higher cell apoptotic rate when compared with that induced by those agents used alone. It is concluded that the bortezomib can induce HL-60 cells apoptosis in a time-and-dose-dependent manner and synergistic effectiveness can be found when bortezomib combined with harringtonine.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drug Synergism ; HL-60 Cells ; Harringtonines ; pharmacology ; Humans ; Pyrazines ; pharmacology