Objective To study the chemical constituents from Clerodendrum inerme.Methods The constituents were isolated and repeatedly purified on silica gel column chromatography and their structures were elucidated by spectral analysis.Results Eight compounds: friedelin (Ⅰ), stigmasterol (Ⅱ), betulinic acid (Ⅲ), acacetin (Ⅳ), syringic acid (Ⅴ), p-methoxybenzoic acid (Ⅵ), apigenin (Ⅶ), and daucosterol (Ⅷ) were isolated from the aerial part of C.inerme. Conclusion Compounds Ⅱ, Ⅲ, Ⅴ, Ⅵ, and Ⅷ are isolated from C.inerme for the first time.

Objective To investigate the optimal window width and window level for measuring the airway dimensions with spiral CT scan in Japanese white big-ear rabbits so as to lay the foundation for airway stenting in animal experiments. Methods Multi-slice spiral CT scanning of cervico-thoracic region was performed in 30 healthy adult Japanese white big-ear rabbits, the anteroposterior and transversal diameter of the thoracic trachea, the anteroposterior diameter of the right and left bronchus were measured with lung window, mediastinum window and special fat window separately. The revealing rate of the tracheal wall and the measuring results in different windows and levels were recorded and compared with the anatomical data. The differences of the relevant data were statistically analyzed. Results With lung window, the tracheal wall was well demonstrated, but the relevant data were smaller than that with mediastinum window. With mediastinum window, the data were bigger and the tracheal wall border appeared blurred. The results obtained with fat window were close to the actual anatomical data. Conclusion For accurately measuring the anteroposterior and transversal diameter of the thoracic trachea in Japanese white big-ear rabbits with multi-slice spiral CT scan, fat window should be adopted, which is helpful for the preparation of tracheal and bronchial stents.

DNA Fingerprinting ; DNA, Mitochondrial ; Homicide ; Humans ; Mitochondria

A large amount of nicotinamide adenine dinucleotide phosphate (NADPH) is required for fatty acid synthesis and maintenance of the redox state in cancer cells. Malic enzyme 1(ME1)-dependent NADPH production is one of the three pathways that contribute to the formation of the cytosolic NADPH pool. ME1 is generally considered to be overexpressed in cancer cells to meet the high demand for increased de novo fatty acid synthesis. In the present study, we found that glucose induced higher ME1 activity and that repressing ME1 had a profound impact on glucose metabolism of nasopharyngeal carcinoma(NPC) cells. High incorporation of glucose and an enhancement of the pentose phosphate pathway were observed in ME1-repressed cells. However, there were no obvious changes in the other two pathways for glucose metabolism: glycolysis and oxidative phosphorylation. Interestingly, NADPH was decreased under low-glucose condition in ME1-repressed cells relative to wild-type cells, whereas no significant difference was observed under high-glucose condition. ME1-repressed cells had significantly decreased tolerance to low-glucose condition. Moreover, NADPH produced by ME1 was not only important for fatty acid synthesis but also essential for maintenance of the intracellular redox state and the protection of cells from oxidative stress. Furthermore, diminished migration and invasion were observed in ME1-repressed cells due to a reduced level of Snail protein. Collectively, these results suggest an essential role for ME1 in the production of cytosolic NADPH and maintenance of migratory and invasive abilities of NPC cells.
Carcinoma ; Cell Line, Tumor ; Cell Movement ; Cell Survival ; Glucose ; metabolism ; Glycolysis ; Humans ; Malate Dehydrogenase ; metabolism ; NADP ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Oxidation-Reduction ; Oxidative Phosphorylation ; Pentose Phosphate Pathway ; Proto-Oncogene Proteins c-akt ; metabolism

The objective was to identify some biochemical and physical properties for fusion protein IL6D24-PE40KDEL. Edman degradation, SDS-PAGE, peptide mass fingerprinting, Western blot and MTT were used for identification of the protein. The results showed that the sequence of N-terminus is Met-Ile-Asp-Lys-Gln-Ile, Met was added because of prokaryotic expression system; Western blot revealed that the purified protein could react with IL6 and PEA antibody. The purified protein IL6D24-PE40KDEL could kill the multiple myeloma cell lines U266 expressing high affinity IL6R, but it could not kill the cell lines CEM which not expressed IL6R; The molecular weight was 58.7 kD measuring by SDS-PAGE; peptide mass fingerprinting (PMF) confirmed that the construction of IL6D24-PE40KDEL was correct. A novel protein by Peptident database in EXPASY web site was identified. In conclusion, IL6D24-PE40KDEL is a new targeting protein with bioactivity of specific killing effect.
ADP Ribose Transferases ; chemistry ; metabolism ; pharmacology ; Amino Acid Sequence ; Blotting, Western ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Exotoxins ; chemistry ; metabolism ; pharmacology ; Humans ; Interleukin-6 ; chemistry ; metabolism ; pharmacology ; Molecular Sequence Data ; Pseudomonas aeruginosa ; genetics ; metabolism ; Recombinant Fusion Proteins ; chemistry ; metabolism ; pharmacology ; Sequence Analysis, Protein ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

AIMTo observe the antagonistic effect of hydroxysafflor yellow A (HSYA) on the platelet activating factor (PAF).

METHODSWashed rabbit platelet (WRP) aggregation and rabbit polymorphonuclear leukocytes (PMNs) aggregation induced by PAF were observed by turbidimetric assay in vitro. The PAF receptor antagonistic effect of HSYA was investigated by radio ligand binding assay (RLBA).

RESULTSIn RLBA the specific binding inhibition effect of HSYA was found to be concentration-dependent in three different [3H]PAF concentrations. In the experiments, WRP aggregation and rabbit PMNs aggregation induced by PAF (9.55 x 10(-10), 9.55 x 10(-6) mol.L-1) were both inhibited by HSYA in a concentration-dependent manner in vitro. The IC50 of HSYA to inhibit WRP and rabbit PMNs aggregation was 0.99 and 0.70 mmol.L-1, respectively.

CONCLUSIONThe PAF receptor binding can be antagonized by HSYA.

Animals ; Carthamus ; chemistry ; Cell Aggregation ; drug effects ; Chalcone ; analogs & derivatives ; isolation & purification ; pharmacology ; In Vitro Techniques ; Male ; Neutrophils ; drug effects ; Plants, Medicinal ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Membrane Glycoproteins ; antagonists & inhibitors ; Quinones ; isolation & purification ; pharmacology ; Rabbits ; Receptors, G-Protein-Coupled ; antagonists & inhibitors

Objective To observe the improving effect in joint function of moderate skeletal fluorosis treated by traditional Chinese medication(main ingredient was Strychnine).MethodsFrom December 2007 to July 2009,120 moderate skeletal fluorosis patients met the inclusion criteria were divided into the treatment group(60 cases)and the control group(60 cases)in the skeletal fluorosis hospital of Xinzhou,the treatment group was given basic treatment and traditional Chinese medication,the control group wa8 given basic treatment and placebo.The treatment lasted 12 weeks,follow up 24 weeks.Before treatment,after treatment 4 weeks,8 weeks,12 weeks,36 weeks,a third party evaluate comprehensive function of both upper and lower limb and joint dysfunction.Results The main effect of both drugs was statistically significance in the scores of the upper forearm in the finger by touching the posterior contralateral ear, upper arm touched by the finger back in the opposite corner subscapularis function, lower limb function and single-joint dysfunction(F values were 4.08,14.32,35.81,13.02, all P<0.05), the main effect of time also was significant (F values were 82.63,72.82,277.33,328.16, all P<0.05),①the upper forearm in the finger by touching the posterior contralateral ear functions:At the time of 8,12 weeks,scores of the treatment group were lower than those of before treatment and control group (all P<0.05);At the time of 36 weeks,scores of the treatment group were lower than that 12 weeks(all P<0.05);At the time of 8,12,36 weeks, scores of the control group were lower than those of before treatment(all P < 0.05);②upper arm function, namely fingers touching the opposite corner subscapularis:At the time of 4,8,12 weeks, scores of the treatment group were lower than those of before treatment(all P<0.05); At the time of 36 weeks, scores of the treatment group were lower than that 12 weeks(all P<0.05); At the time of 8,12,36 weeks, scores of the treatment group were lower than those of the control group (all P<0.05);③Lower extremity functions: At the time of 8,12 weeks,scores of the treatment group were lower than those of before treatment and control group (all P<0.05);At the time of 36 weeks, scores of the treatment group were lower than that 12 weeks(all P<0.05) ; At the time of 8,12,36 weeks, scores of the control group were lower than those of before treatment (all P<0.05);④single joint functions:At the time of 4,8,12 weeks,scores of the treatment group were lower than those of before treatment(all P<0.05); At the time of 36 weeks,scores of the treatment group were lower than that 12 weeks(all P<0.05) ; At the time of 8,12,36 weeks, scores of the control group were lower than those of before treatment(all P<0.05);At the time of 4,8,12,36 weeks, scores of the treatment group were lower than those of control group(all P<0.05);⑤At the end of treatment and follow-up,the improvement rate in joint functions in the treatment group were 88.33% (53/60),93.33% (56/60); the control group were 28.07%(16/57),40.35%(23/57), (Fisher test, P<0.01,X2=56.21, P<0.01). ConclusionTraditional Chinese medication(its main ingredient is Strychnine), an effective drug for improving joint dysfunction in patients suffering from moderate skeletal fluorosis, is simple and effective.

The objective of this study was to investigate the efficacy and toxicity of standard-dose idarubicin in combination with continuous infusion of cytarabine as induction therapy in patients with acute myeloid leukemia (AML). A total of 38 AML patients were enrolled, including 30 new diagnosed patients, 8 relapsed and refractory patients. Cytogenetic analysis was performed in all patients, 15 patients had cytogenetic aberrations including 4 complex abnormalities. All patients were treated with standard-dose idarubicin [12 mg/(m(2).d), days 1 to 3] and continuous infusion of cytarabine [100 mg/(m(2).d), days 1 to 7]. The results showed that after one course of induction therapy, the overall response rate was 89.5% (34/38), and 32 out of 38 (84.2%) patients achieved complete remission (CR), including 27 of 30 (90.0%) new diagnosed AML patients, 5 (62.5%) refractory and relapsed AML patients, all 4 patients with complex cytogenetic aberrations achieved cytogenetic CR. Out of 6 relapsed patients 2 showed as extramedullary relapse, 4 showed as bone marrow relapse. The median survival duration was > 22 months and median disease-free survival time was > 16 months. Myelosuppression and infections due to neutropenia were the most frequent adverse effects, severe nonhematologic toxicity and the early death were not observed in the patients. It is concluded that standard-dose of idarubicin combined with continuous infusions of cytarabine as the induction therapy is highly effective and well tolerated approach in patients with AML, this regimen provides an opportune moment for hematopoietic stem cell transplantation.
Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Cytarabine ; administration & dosage ; therapeutic use ; Female ; Humans ; Idarubicin ; administration & dosage ; therapeutic use ; Leukemia, Myeloid, Acute ; drug therapy ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the intrinsic connection between activation of classical nuclear factor-κB (NF-κB) pathway and gefitinib resistance in human lung adenocarcinoma H1650 cells.

METHODSHuman lung adenocarcinoma H1650 cells were exposed to gefitinib continuously for 60 days to obtain resistant H1650 cells. The expressions of P-IκBα, P-p50 and P-p65 in the cytoplasm or nuclei were detected using Western blotting in human lung adenocarcinoma HCC827 cells, parental H1650 cells and gefitinib-resistant H1650 cells. The effects of gefitinib alone or in combination with PDTC on the survival rate and expressions of NF-κB P-p50 and P-p65 were compared among the 3 cell lines.

RESULTSGefitinib-resistant H1650 cells showed increased cytoplasmic and nuclear P-IκBα expressions. The expressions of P-p50 and P-p65 differed significantly among the 3 cell line, decreasing in the order of resistant H1650 cells, parental H1650 cells, and gefitinib sensitive HCC827 cell lines (P<0.05 or 0.01). Treatment with gefitinib alone resulted in a significantly lower cell inhibition rate in resistant H1650 cells than in the parental H1650 cells (P<0.05) and HCC827 cells (P<0.01). The resistant H1650 cells had a significantly higher expression of P-p50 and P-p65 than other two cell lines (P<0.05). In both the resistant and parental H1650 cells, gefitinib significantly lowered P-p50 and P-p65 expressions (P<0.05 or 0.01), and the combined treatment with gefitinib and PDTC significantly decreased the cell survival rate and further lowered the cytoplasmic and nuclear expressions of P-p50 and P-p65 (P<0.01 or 0.01).

CONCLUSIONThe activation of classical NF-κB pathway is a key factor contributing to transformation of the parental H1650 cells into gefitinib-resistant cells. Gefitinib combined with PDTC can inhibit P-IκBα production and NF-κB P-p50 and P-p65 activation to suppress the survival of residual H1650 cells and the generation of gefitinib-resistant cells.

OBJECTIVETo study the expression of recombinant human SCF-TPO fusion protein and its biological function.

METHODSFour primers were designed according to known sequences of TPO and SCF. The functional amino acid domains of TPO and SCF were amplified by RT-PCR from fetus hepatocytes, respectively. The expression plasmid pET32a/SCF-TPO was constructed by VOE gene fusion technique and expressed in E. coli BL21(DE3) plysS as inclusion body after isopropyl-beta-D-1-thiogalactopyranoside (IPTG) induction. The fusion protein was tested by SDS-PAGE and Western blot. The biological functions of SCF-TPO fusion protein in MO7e cells was investigated by MTT method after purification with metal chelating chromatography.

RESULTSThe high expression SCF-TPO fusion protein was obtained, reaching up to 30% of the total cellular protein. Western blot verified the correct fusion expression and MTT results showed the growth promoting effect of the SCF-TPO fusion protein on MO7e cells, with a higher promoting activity at 100 ng/ml.

CONCLUSIONSExpressed SCF-TPO fusion protein after renaturation has biological activity in promoting the proliferation of MO7e cells.

Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; genetics ; physiology ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Genetic Vectors ; Humans ; Recombinant Fusion Proteins ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cell Factor ; genetics ; metabolism ; physiology ; Thrombopoietin ; genetics ; metabolism ; physiology