OBJECTIVETo investigate synergistically killing effect of histone deacetylase (HDAC) inhibitor suberoylanilide hydroxamic acid (SAHA) combined with imatinib on human chronic myeloid leukemia (CML) cell line.

METHODSK562 cells were co-treated with SAHA and imatinib. Cell growth was measured by MTT assay. Apoptosis was determined using Hoechst staining apoptosis detection kit and flow cytometric analysis. Activation of Caspase pathway, expression of Bcr-Abl and its downstream target genes, and expression of anti-apoptotic proteins were detected by Western blot.

RESULTSSAHA synergized the cytotoxicity of imatinib against leukemia K562 cells, concomitantly with increased apoptosis and enhanced activation of Caspase-3, -8 and PRAP. The combination therapy resulted in significantly lower levels of Bcr-Abl,phosphorylated Bcr-Abl compared to treatment with either SAHA or imatinib alone. Furthermore,the co-treatment resulted in down-regulation of anti-apoptotic protein Mcl-1 expression. Also,marked down-regulated expression of JAK2,STAT5,and phosphorylated STAT5 was detected in the combination therapy.

CONCLUSIONCombining HDAC inhibitor SAHA with imatinib can kill CML cells synergistically by inhibiting cell growth and inducing apoptosis, which is associated with activation of Caspase pathway and regulation of anti-apoptotic proteins.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Benzamides ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Drug Synergism ; Fusion Proteins, bcr-abl ; metabolism ; Histone Deacetylase Inhibitors ; pharmacology ; Humans ; Hydroxamic Acids ; pharmacology ; Imatinib Mesylate ; Intracellular Signaling Peptides and Proteins ; metabolism ; Janus Kinase 2 ; metabolism ; K562 Cells ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology ; STAT5 Transcription Factor ; metabolism

In order to explore the dormancy physiological and biochemical mechanism of Paris seeds, the seed embryo growth courses, and the dynamic change of 5 enzymes, include SOD, POD, CAT, MDH, G-6-PDH were measured during variable temperature stratification. The results indicated that Paris seeds embryo grew quickly after 40 d in warm-stratification (18 ± 1) °C, at the meantime the metabolic activity was significantly strengthened. These facts showed that Paris seeds turned into physiological after-ripening process. After 60-80 d, the morphological embryo after-ripping process basically completed, and the following cold-stratification (4 ± 1) °C furthered Paris seed to finish physiological after-ripening. After 40 d, the activity of MDH decreased while G-6-PDH increased significantly. This showed that the main respiratory pathway of seed changed from TCA to PPP, which benifited breaking seed dormancy. In the whole period of stratification process, the activity variation of SOD and CAT was insignificantly and the activity of POD was enhanced significantly after shifting the seed in cold stratification process. This showed that SOD, CAT had no direct effects on breaking Paris seed dormancy but keeping the seed vigor, while the POD might involve in the process of Paris seed dormancy breaking.
Germination ; Liliaceae ; chemistry ; embryology ; enzymology ; Plant Proteins ; metabolism ; Seeds ; chemistry ; enzymology ; growth & development ; Temperature

OBJECTIVEIt has been shown that triptolide can attenuate pulmonary arterial hypertension in rats. This study was designed to investigate the therapeutic effect of triptolide on pulmonary hypertension in rats and possible mechanisms.

METHODSSixty Sprague-Dawley (SD) rats were randomly divided into 6 groups: normal control, model, continuous triptolide-treated, delayed triptolide-treated and two placebo groups for continuous and delayed fashions (n=10 each). The rats from the last 5 groups were injected with monocrotaline (MCT, 60 mg/kg) on day 7 after left pneumonectomy. The rats in the continuous triptolide-treated group received therapy from day 5 to 35 with triptolide (0.25 mg/kg intraperitoneally, every other day) and those in the delayed triptolide-treated received therapy with triptolide (0.20 mg/kg intraperitoneally, daily) from day 21 to 35 after operation. The hemodynamic parameters were detected by catheterization and the pathologic changes of small pulmonary arteries were evaluated by light microscopy 5 weeks post-operation. The expression of matrix metalloproteinases (MMPs) was assessed by immunohistochemistry and quantitative fluorescence PCR of relevant (MMP2 and MMP9) mRNAs.

RESULTSBy day 35 after operation, the mean pulmonary arterial pressure (mPAP, 38.10+/-1.20 vs 16.70+/-1.16 mmHg)the ratio of right ventricle/left ventricle plus septum [RV/(LV+S), 62.45+/-5.28% vs 22.76 +/-3.01%] and the vessel obstructive scores (VOS, 1.736 +/-0.080 vs 0.000 +/-0.000) increased significantly in the Model group compared with those of the normal control group (P < 0.01). The expression of MMP2 and MMP9 and their mRNA expression in lung tissues obviously also elevated in the Model group (P < 0.05). The continuous and the delayed triptolide-treated groups had significantly lower mPAP (20.80+/-1.03 and 26.20+/-1.03 mmHg, respectively) and less right ventricular hypertrophy and pulmonary arterial neointimal formation compared with the model and the placebo groups. The two treated groups also demonstrated decreased expression of MMP2 and MMP9 and their mRNA expression in lung tissues. There were significant differences in mPAP, RV/(LV+S) and VOS between the two triptolide-treated groups.

CONCLUSIONSTriptolide attenuates the development of pulmonary hypertention and right ventricular hypertrophy and promotes regression of pulmonary arterial neointimal formation in pneumonectomized rats that received MCT, possibly through an inhibition of MMPs activity.

Animals ; Diterpenes ; pharmacology ; therapeutic use ; Epoxy Compounds ; pharmacology ; therapeutic use ; Hypertension, Pulmonary ; drug therapy ; enzymology ; Immunohistochemistry ; Lung ; enzymology ; Male ; Matrix Metalloproteinase 2 ; analysis ; genetics ; Matrix Metalloproteinase 9 ; analysis ; genetics ; Phenanthrenes ; pharmacology ; therapeutic use ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley

BACKGROUND:As the regulators of cytokines, suppressors of cytokine signaling (SOCS) play an important role in the inflammation reaction. Some studies found that SOCS-1 and SOCS-3 were involved in the pathogenesis of some inflammatory diseases such as rheumatoid arthritis, inflammatory bowel disease. But the expressions of SOCS in coronary heart disease have not yet been reported. This study aimed to investigate the expression and clinical significance of SOCS-1 and SOCS-3 in the myocardium of patients with sudden cardiac death (SCD).METHODS:Myocardial autopsy specimens were collected from 24 patients at the Forensic Medicine Department of Sun Yat-Sen University, Guangzhou, China between 2005 and 2006. Of them, 9 patients had autopsy findings consistent with coronary atherosclerosis (non-myocardial infarction) leading to SCD (non-MI group), 7 died of acute myocardial infaction (MI group), and 8 died from traffic accidents and trauma (control group). The expressions of SOCS-1 mRNA and SOCS-3 mRNA in the myocardium of the non-MI, MI and control groups were detected using RT-PCR. The levels of SOCS-1 and SOCS-3 proteins were detected using immunohistochemistry. Statistical analyses were performed using SPSS version 13.0 software and the data were analyzed by ANOVA.RESULTS:The expressions of SOCS-1 mRNA and SOCS-3 mRNA in the non-MI and MI groups were significantly higher than those in the control group[(0.788±0.101), (0.741±0.111) vs. (0.436±0.044), (P<0.01); (0.841±0.092), (0.776±0.070) vs. (0.454±0.076), (P<0.01)] respectively. The antibody-positive cells of SOCS-1 protein in the myocardium of the non-MI and MI groups were significantly higher than those in the myocardium of the control group[(320.00±48.48), (347.14±70.88) vs. (42.50±10.35), (P<0.01)] respectively. The antibody-positive cells of SOCS-3 protein in the myocardium of the non-MI and MI groups were significantly higher than those in the myocardium of the control group[(381.11±59.25) vs. (40.00±10.69), (P<0.01)] and[(332.86±111.91) vs. (40.00±10.69), (P=0.001)].CONCLUSION:The expressions of SOCS-1 and SOCS-3 in the myocardium of patients with SCD from coronary heart disease are significantly increased and contribute to the pathogenesis of SCD.

Aged ; Female ; Femoral Neck Fractures ; diagnostic imaging ; surgery ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Radiography ; Retrospective Studies ; Treatment Outcome

Animals ; Breath Tests ; methods ; Carbon Isotopes ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Chemical and Drug Induced Liver Injury ; etiology ; physiopathology ; Liver Function Tests ; methods ; Male ; Phenylalanine ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo compare therapeutic effects of local injection with autoallergic platelet rich plasma (PRP) or autoallergic whole blood (AWB) for the treatment of chronic tennis elbow.

METHODSFrom January 2011 to January 2014, 40 patients with chronic tennis elbow were divided into 2 groups, 20 cases in each group: PRP group and AWB group. There were 20 patients in PRP group treated with local injection of autoallergic platelet rich plasma, including 5 males and 15 females, with an average age of (47.50 ± 9.86) years old; and the average course of disease was (4.67 ± 3.27) months. Among the 20 patients in AWB group treated with local injection of autoallergic whole blood, 3 patients were male and 17 patients were female, with an average age of (46.50 ± 9.96) years old;and the average course of disease was (4.53 ± 2.27) months. The elbow joint was fixed with elastic stockings after injection. All the patients were guided to do strengthening and extension exercises during the follow-up period. Visual analog scale (VAS), Mayo scores for elbow and pressure pain threshold (PPT) were used to evaluate clinical effects after injection immediately and 4,8 weeks after treatment. Results:All the patients were followed up,there were no infections and swelling occurred. The VAS, Mayo and PPT scores of patients in PRP group were improved from pre-therapy 7.22 ± 1.32, 56.71 ± 10.90 and 17.47 ± 4.62 to 2.73 ± 1.00, 91.59 ± 6.95 and 21.35 ± 4.80 respectively 8 weeks after treatment. The VAS, Mayo and PPT scores of patients in AWB group were improved from pre-therapy 7.16 ±1.27, 54.72 ± 8.36 and 17.06 ± 4.83 to 3.81 ± 1.36, 82.06 ± 7.89 and 20.12 ± 4.97 respectively 8 weeks after treatment. All the pain and functional variables including VAS, PPT, and Mayo scores were improved significantly in both groups 4 weeks after injection. On the 4th week after injection, there was no statistically significant difference in PPT between two groups; while the VAS and Mayo score of AWB group were lower than those of PRP group. On the 8th week after injection, the VAS of AWB group was higher than that of PRP group; but the Mayo and PPT scores of AWB group were lower than those of PRP group.

CONCLUSIONPRP and AWB injections are both effective to treat chronic lateral epicondylitis. Compared with AWB injection, PRP injection may be more effective in releasing pain and improving function for a longer time.

Adult ; Blood Transfusion ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Pain Threshold ; Platelet-Rich Plasma ; Tennis Elbow ; therapy ; Visual Analog Scale

This study was amid to construct the pharmacophore model of L-type calcium channel antagonist in the application of screening Drugbank and TCMD. This paper repositions the approved drugs resulting from virtual screening and discusses the relocation-based drug discovery methods, screening antihypertensive drugs with L-type calcium channel function from TCMD. Qualitative hypotheses wre generated by HipHop separately on the basis of 12 compounds with antagonistic action on L-type calcium channel expressed in rabbit cardiac muscle. Datebase searching method was used to evaluate the generated hypotheses. The optimum hypothesis was used to search Drugbank and TCMD. This paper repositions the approved drugs and evaluates the antihypertensive effect of the chemical constituent of traditional Chinese medicine resulting from virtual screening by the matching score and literature. The results showed that optimum qualitative hypothesis is with six features, which were two hydrogen-bond acceptors, four hydrophobic groups, and the CAI value of 2.78. Screening Drugbank achieves 93 approved drugs. Screening TCMD achieves 285 chemical constituents of traditional Chinese medicine. It was concluded that the hypothesis is reliable and can be used to screen datebase. The approved drugs resulting from virtual screening, such as pravastatin, are potentially L-type calcium channels inhibitors. The chemical constituents of traditional Chinese medicine, such as Arctigenin III and Arctigenin are potentially antihypertensive drugs. It indicates that Drug Repositioning based on hypothesis is possible.
Animals ; Antihypertensive Agents ; chemistry ; pharmacology ; Calcium Channel Blockers ; chemistry ; pharmacology ; Calcium Channels, L-Type ; genetics ; metabolism ; Drug Repositioning ; methods ; Molecular Structure ; Myocardium ; metabolism ; Rabbits

OBJECTIVETo design DNA microarray and investigate the molecular anti-tumor mechanism of herbs of traditional Chinese medicine.

METHODcDNA microarrays consisting of 56 probes representing 24 human cell cycle genes were constructed, Four anti-hepatocarcinoma herbs including Radix Linderae, Hebra Artemisiae Annuae, Radix Amebiae, Radix Astragli, were chosen. Effects of herbs on SMMC-7721 cell cycle were observed by flow cytometry assay. Effects of herbs on cell cycle gene expression in SMMC-7721 cells were analyzed by comparing hybridization of Dig-Labeled cDNAs from herb-treated cells and cDNAs from untreated cells.

RESULTExpressions of cell cycle geneswere changed in different degrees after herbs treated. Some genes were down-regulated and some genes were up-regulated. The changes in gene expression agreed with the results of flow cytometry assay.

CONCLUSIONThe results suggest that these herbs may have effects on cell cycle and DNA damage checkpoint genes which may be the mechanism of the herbs, and DNA microarray can be used to investigate the biological function of extracts of traditional Chinese medicine.

Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Artemisia ; chemistry ; Astragalus membranaceus ; chemistry ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Cyclin-Dependent Kinases ; genetics ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Gene Amplification ; Gene Expression Profiling ; Genes, cdc ; drug effects ; Humans ; Lindera ; chemistry ; Lithospermum ; chemistry ; Liver Neoplasms ; metabolism ; pathology ; Oligonucleotide Array Sequence Analysis ; methods ; Plants, Medicinal ; chemistry ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; cdc25 Phosphatases ; genetics ; metabolism

Construction of mutant strain is an essential method in pathogenesis researches. The conventional method for Brucella unmarked deletion mutant construction is based on suicide plasmid, but the efficiency is very low. In the present study, we first optimized the electroporation parameters, and then, the cloning plasmid pEX18Gm containing sacB was successfully used to construct unmarked deletion mutant of the type IV secretion system. This indicated that by using conventional cloning plasmid as suicide plasmid in Brucella, unmarked deletion mutants can be constructed with high efficiency.