A UPLC method has been developed in the current investigation for simultaneous determination of nine chemical markers of Bletilla striata, 4-hydroxymethylphenyl beta-D-glucoside, blestroside, dactylorhin A, militarine, dihydrophenanthrene 5, gymnoside V, dihydrophenanthrene 1, benzylphenanthrene 3 and gymnosides IX. Separation was performed at 45 degrees C on an ACQUITY UPLC BEH C18 column (2.1 mm x 150 mm, 1.7 microm) with a gradient solvent system of acetonitrile-water as the mobile phase. The flow rate was 0.3 mL x min(-1), the detection wavelength was 280 nm. The results showed that the nine chemical markers could be well resolved and that in the selected linear range, all calibration curves of the nine chemical markers showed good linearity (r > or = 0.999 3). The recoveries (n = 6) were in the range of 98.15% - 102.2% and RSDs were between 2.1% - 3.6%. The data suggested that the developed UPLC-UV method had good reproducibility, robustness, and accuracy, which was suitable for the quality control of Bletilla striata. Applications of the method showed that the nine chemical markers had higher contents in the wild B. striata than in the cultivated ones.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Magnoliopsida ; chemistry ; Rhizome ; chemistry

Objective To discuss the CT manifestations of multi-parasite infection in liver to improve the diagnostic accuracy. Methods A total of 11 patients with ELISA-proved multi-parasite infection in liver were enrolled in this study.The plain and dynamic enhanced CT scans were performed.The imaging findings including the number,distribution,size,shape,density,enhancement and degree intratumaral features,cholangiectasis and abdominal lymphadenopathy were retrospectively analyzed.Results A single lesion of multi-parasite infection in liver was detected in 6 patients and multiple lesions were in other 5.The lesions in 8 patients were located in a single lobe of the liver,and involved in several hepatic lobes in other 3.The maximum diameter of the lesion ranged from 1.7 cm to 6.3 cm with an average diameter of 4.4 cm.Irregular lesions were found in 7 patients and round ones in shape were in other 4.Plain CT showed the lesions with low-intensity.All lesions were mild to moderate inhomogeneous enhancement on portal vein phase,presented honeycomb-like or separated enhancement with cholangiectasis (n=2)and abdominal lymphadenopathy (n=2).Conclusion The CT manifestations on portal vein phase in combination with clinical data are useful for the diagnosis of multi-parasite infection in liver.

OBJECTIVETo study the expressions of gastric mucosal proteins in chronic gastritis (CG) rats of Pi-Wei damp-heat syndrome (PWDHS), to investigate the pathogenesis correlated to CG rats of PWDHS, to observe the differential expressions of gastric mucosal proteins in CG rats of PWDHS, and to investigate the mechanisms of Sanren Decoction (SD) for treating CG rats of PWDHS.

METHODSTotally 36 male SD rats were adaptable fed for 3 days and randomly divided into 3 groups, i.e., the normal control group, the CG of PWDHS rat model group (abbreviated as the model group), and the SD treatment group, 12 in each group. The CG of PWDHS rat model was prepared by composite factors. The gastric mucosal protein was separated using two-dimensional gel electrophoresis technique, and stained by Coomassie brilliant blue. The protein spots expressed differently were analyzed by PDquest 8.0 software. The protein spots expressed differently was identified by MALDI-TOF/TOF-MS.

RESULTSThe protein spots were 1 025 +/- 3 9, 994 +/- 51, 1 087 +/- 33 in the normal control group, the model group, and the SD treatment group respectively detected from two-dimensional gel electrophoresis profiles. Compared with the normal control group, there were 74 protein spots differentially expressed in the model group, 30 spots up-regulated and 44 spots down-regulated. Compared with the model group, there were 75 protein spots differentially expressed in the SD treatment group, 49 spots up-regulated and 26 spots down-regulated. Five protein spots differentially expressed were successfully identified, i.e., heat shock protein 72 (HSP72), heat shock protein 60 (HSP60), protein disulfide-isomerase (PDI), malate dehydrogenase (MDH), and unnamed protein.

CONCLUSIONSThe pathogenesis of CG of PWDHS may be correlated to energy metabolism disturbance and stress. The mechanisms of SD for treating it may possibly adjust differential expressions of gastric mucosal proteins.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; metabolism ; Gastritis ; diagnosis ; drug therapy ; metabolism ; Male ; Medicine, Chinese Traditional ; Phytotherapy ; Proteome ; metabolism ; Rats ; Rats, Sprague-Dawley

Animals ; Apoptosis ; drug effects ; Cell Line ; Cell Proliferation ; Curcuma ; Extracellular Matrix ; metabolism ; Hepatic Stellate Cells ; drug effects ; pathology ; Plant Extracts ; pharmacology ; Plant Oils ; pharmacology ; Rats

This study was purpose to investigate the role of reactive oxygen species (ROS) in apoptosis and autophagy induced by FTY720 in multiple myeloma cell line U266. U266 cells were treated by different concentrations of FTY720 for 24 h, the apoptotic rates were detected by flow cytometry, and the expression of LC3B was detected by Western blot. The results indicated that apoptosis and autophagy were induced by FTY720 in U266 cells. Autophagy induced by FTY720 could lead to cell death. Bafilomycin A1, the inhibitor of autophagy, could enhance the cell viability in U266 cells treated with FTY720. NAC or Tiron, ROS scavenger, could decrease the FTY720 induced apoptosis and the expression of LC3B-II was reduced in combination of FTY720 with NAC or Tiron as compared with treatment with FTY720 only. It is concluded that FTY720 can induce U266 cell apoptosis and autophagy. ROS is the mediator that regulates both the apoptosis and autophagy in multiple myeloma cells.
1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt ; Apoptosis ; drug effects ; Autophagy ; drug effects ; Cell Line, Tumor ; Fingolimod Hydrochloride ; Humans ; Macrolides ; Microtubule-Associated Proteins ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; Propylene Glycols ; pharmacology ; Reactive Oxygen Species ; metabolism ; Sphingosine ; analogs & derivatives ; pharmacology

This study was aimed to investigate the influence of TIEG1 on apoptosis of HL-60 cells and the expression of Bcl-2/Bax. Different concentration of TIEG1 were used to treat HL-60 cells, the cell growth inhibition rate was detected by MTT method. After treating HL-60 cells with 12.03 ng/ml TIEG1, cell apoptosis was detected with flow cytometry. Bcl-2 and Bax was detected with RT-PCR. The results showed that TIEG1 had inhibitory effect on HL-60 cell proliferation, and in time-and dose-dependent manners. The more obvious inhibitory effect was observed in HL-60 cells treated with TIEG1 of 12.03 ng/ml. During the course of cell apoptosis, Bax expression increased, but Bcl-2 expression decreased (P < 0.05). It is concluded that TIEG1 inhibits HL-60 cell proliferation and induces apoptosis in time and dose-dependent manners. During the course of HL-60 cells apoptosis induced by TIEG1, Bcl-2/Bax are associated with HL-60 cell apoptosis induced by TIEG1.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Early Growth Response Transcription Factors ; pharmacology ; Gene Expression Regulation, Leukemic ; HL-60 Cells ; Humans ; Kruppel-Like Transcription Factors ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

This study was aimed to investigate the effects of valproic acid sodium (VPA) on the proliferation and apoptosis of acute T-lymphoblastic leukemia Jurkat cells. Jurkat cells were treated with different concentration of VPA. Proliferation-inhibition curve was assayed and plotted by CCK-8 method and the cell apoptosis was detected by flow cytometry with Annexin V/PI double staining. The expression level of anti-apoptotic gene BCL-2 and pro-apoptosis gene Bak1 were detected by semi-quantitative RT-PCR. The results showed that the VPA inhibited the proliferation of Jurkat cells in concentration-dependent manner. As compared with the control group, the apoptosis of cells increased along with adding concentration of VPA; VPA could decrease the expression of BCL-2 gene, but did not show obvious effect on the expression of Bak1. It is concluded that the VPA can inhibit proliferation of Jurkat cells which possibly associates with the decrease of BCL-2 expression.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Jurkat Cells ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Sodium ; pharmacology ; Valproic Acid ; pharmacology ; bcl-2 Homologous Antagonist-Killer Protein ; metabolism

This study was aimed to explore the effects of oleanolic acid on PTEN expression and apoptosis of Jurkat cells. The inhibitory rate was measured by Cell Counting Kit-8. The apoptotic nucleus morphous was observed by Hoechst 33258 staining. The apoptosis rate of Jurkat cells were determined by flow cytometry with Annexin V/PI double staining. PTEN mRNA and protein were detected by quantitative real-time PCR and Western blot respectively. The results showed that oleanolic acid inhibited the proliferation of Jurkat cells in time- and dose-dependent manners. The 50% growth inhibition (IC(50)) at 12, 24 and 48 hours were about 85.35 µmol/L, 53.66 µmol/L and 33.18 µmol/L respectively. Flow cytometric assay showed that the apoptotic rates of Jurkat cells treated with oleanolic acid (0, 40, 80 and 160 µmol/L) for 24 hours were 6.72%, 19.8%, 28.72% and 30.12% (p < 0.05). PTEN mRNA and protein expressions were up-regulated in Jurkat cells treated with oleanolic acid of concentration 80 µmol/L and 160 µmol/L for 24 hours. It is concluded that up-regulation of PTEN mRNA and PTEN protein may be involved in oleanolic acid-induced Jurkat cell apoptosis.
Apoptosis ; drug effects ; Cell Proliferation ; Humans ; Jurkat Cells ; Oleanolic Acid ; pharmacology ; PTEN Phosphohydrolase ; metabolism ; Up-Regulation

This study was aimed to investigate the relationship of Ki-67 proliferation index (Ki-67 PI) with non-Hodgkin's lymphoma(NHL) typing and biological behavior, as well as its significance in clinical characters and prognosis of diffuse large B-cell lymphoma(DLBCL). A total of 542 cases of NHL in our hospital from 1st January 2001 to 31st December 2010 were retrospectively analyzed, and Ki-67 PI was all assayed immunohistochemically, and a total of 82 cases of newly-diagnosed DLBCL with more clinical records were investigated. The results indicated that according to the World Health Organization (WHO) histopathological classification of lymphoma, Ki-67 PI was different as classification for NHL subgroups was different. The Ki-67 PI increased with aggressive progression of NHL. The mean Ki-67 PI ranged from 25.5% in indolent lymphoma to 98.4% in very aggressive lymphoma. ROC curve analysis demonstrated that the 50% was the cut-off value distinguishing indolent from aggressive disease. On ROC curve analysis, Ki-67 PI of 75% was found to significantly discriminate patients with DLBCL who had a good or bad prognosis. There was a significant correlation of Ki-67 PI with Ann Arbor stage and LDH level. When the DLBCL cases were divided by Ann Arbor stage and IPI score, the 3-year overall survival (OS) of patients with a low Ki-67 PI (≤ 75%) in the group of Ann Arbor stage III-IV and high LDH level was higher than those with a high Ki-67 PI (> 75%) among the patients with B symptoms and IPI 3.0-5. 3-year OS in those with a low Ki-67 PI (≤ 75%) in the group of Ann Arbor stage III-IV and normal LDH level was higher than those with a high Ki-67 PI (> 75%) among the patients with B symptoms. 3-year OS of patients with a low Ki-67 PI (≤ 75%) in the group at III-IV stage and a high LDH level was higher than those with a high Ki-67 PI (> 75%). It is concluded that a cut-off value of 50% can be helpful to differentiate indolent from aggressive NHL. In DLBCL, a cut-off value of 75% can distinguish patients with a good or bad prognosis when combined with other prognostic factors, i.e. B symptoms, Ann Arbor stage, IPI score and lactate dehydrogenase (LDH) level.
Female ; Humans ; Ki-67 Antigen ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Lymphoma, Non-Hodgkin ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis

The aim of this study was to investigate the effect of proteasome inhibitor bortezomib on the expression of ERK, JNK, and P38 in daunorubicin (DNR)-resistant K562 cells and its mechanism. MTT method was used to determine the drug-resistant K562 cells and the cellular toxicity of bortezomib; Western blot was used to detect the expression of protein ERK, JNK and P38 in K562 cells after treatment with 100 nmol/L DNR alone or combined with 1 nmol/L and 10 nmol/L bortezomib for 36 hours. Flow cytometry assay was used to detect the apoptosis rate in each group cells. The results indicated that the expression of ERK and P38 were significantly suppressed (p < 0.05) and the expression of JNK was significantly enhanced (p < 0.05) in the cells treated by DNR combined with bortezomib. It is concluded that bortezomib can decrease the expressions of protein ERK and P38 and enhance the expression of JNK, the bortezomib reverses the cellular drug-resistance and promote cell apoptosis through MAPK pathway.
Antineoplastic Agents ; administration & dosage ; pharmacology ; Boronic Acids ; administration & dosage ; pharmacology ; Bortezomib ; Drug Resistance, Neoplasm ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; K562 Cells ; Protease Inhibitors ; administration & dosage ; pharmacology ; Pyrazines ; administration & dosage ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; metabolism