To study the chemical constituents of Abacopteris penangiana (Hook.) Ching, various chromatographic techniques were used to isolate and purify the constituents. The structures of the obtained compounds were elucidated by spectroscopic data and physical-chemical properties. Two compounds were isolated from the n-BuOH soluble fraction of an acetone-H2O (4:1) extract of A. penangiana and were identified as 4'-hydroxy pneumatopterin B (I) and 6"-O-acetyl triphyllin A (II). Compounds I and II are new compounds.
Ferns ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Molecular Structure ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry

BACKGROUNDJaw osteonecrosis possibly associated with the administration of bisphosphonates is expected to be treated with a non-pharmacologic approach. This study aimed to determine whether noninvasive, mechanically mediated vibration would inhibit the decline in bone mineral density (BMD) that follows menopause, enhance the BMD of the lumbar and femoral neck, and reduce chronic back pain in postmenopausal women with osteoporosis.

METHODSA total of 116 postmenopausal women with osteoporosis participated in this study, and they were divided into groups A (66 patients) and B (50). Group A received vibration treatment (Subjects vertically stand on the vibration platform, with a vibration frequency of 30 Hz, amplitude of 5 mm; they received the treatment five times per week, ten minutes each time and totally for six months), whereas women of group B served as controls without any treatment. L2 - 4 BMD, bilateral femoral neck BMD, and body mass index (BMI) were recorded before the treatment or at the third and sixth months of the treatment respectively. After the ending of the treatment, the change of BMD in each group was compared and analyzed. Chronic back pain was evaluated by visual analogue scale (VAS) at baseline and the third and sixth months of the treatment.

RESULTSOf the 116 women, 94 including 51 women from group A ((61.23 +/- 8.20) years) and 43 women from group B ((63.73 +/- 5.45) years), completed the study. There were no significant differences in baseline characteristics including age, BMI, menopausal years, lumbar BMD, femoral neck BMD, and VAS between the two groups. The lumbar BMD of the 51 women in group A increased by 1.3% (P = 0.034) after vibration treatment for 3 months and by 4.3% at the sixth month (P = 0.000). The lumbar BMD in group B was decreased at the third month, but there was not statistical significance (P > 0.05). At the sixth month, it was decreased by 1.9% (P < 0.05). The femoral neck BMD of the 51 women in group A was slightly increased after vibration treatment for 3 months, but without statistical significance (P > 0.05). At the sixth month, the BMD was increased by 3.2% (P < 0.05). In group B, the BMD was not decreased significantly (P = 0.185) at the third month, but decreased significantly at the sixth month (1.7%) (P < 0.05) compared with the baseline. Chronic back pain (VAS) reduced more significantly in group A at the third and the sixth months (P < 0.05) after vibration therapy in comparison with the baseline. The BMI was not significantly changed in the two groups during the period of follow-up.

CONCLUSIONSVibration therapy appears to be useful in reducing chronic back pain and increasing the femoral neck and lumbar BMD in postmenopausal women with osteoporosis.

Aged ; Back Pain ; prevention & control ; Bone Density ; Female ; Femur Neck ; Humans ; Lumbar Vertebrae ; Middle Aged ; Osteoporosis, Postmenopausal ; therapy ; Vibration ; therapeutic use

OBJECTIVETo study the chemical constituents of the leaf of Isatis indigotica.

METHODChromatography and spectral analysis were respectively used to isolate and identify the constituents.

RESULTThree compounds were isolated from the ethanol extracts of theleaf of I. indigotica, and identified as indirubin, tryptanthrin and L-pyroglutamic acid.

CONCLUSIONL-pyroglutamic acid was isolated from the genus for the first time, and tryptanthrin was isolated from the leaf of this plant for the first time.

Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Indoles ; chemistry ; isolation & purification ; Isatis ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Pyrrolidonecarboxylic Acid ; chemistry ; isolation & purification ; Quinazolines ; chemistry ; isolation & purification

OBJECTIVETo explore the liver-toxic fraction in Rhigoma of Dioscorea bulbifera.

METHODThe rats were randomized into four groups: control group (20% PVP-water), T001(10% total methanol extraction), F002(5% chloroform fraction) and F003(5% methanol fraction). Direct bilirubin (DBil) and Glutamic-pyruvic transaminase (GPT) were examined, and liver index was measured. The histological and morphological observations were performed with optical and electrical microscope.

RESULTT001 and F002 showed significant liver toxicity.

CONCLUSIONThe chloroform fraction was the liver-toxic fraction of D. bulbifera.

Alanine Transaminase ; blood ; Animals ; Bilirubin ; blood ; Chemical and Drug Induced Liver Injury ; blood ; etiology ; pathology ; Dioscorea ; chemistry ; Drugs, Chinese Herbal ; toxicity ; Female ; Liver ; pathology ; ultrastructure ; Male ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study chemical constituents of Arachniodes rhomboidea.

METHODSilica gel column chromatography and Sephadex LH -20 gel column chromatography were employed for the isolation and purification. The structures were identified on the basis of spectral data and chemical methods.

RESULTSix compounds were isolated and identified as follows: kaempferol (1), kaempferol-3-O-alpha-L-rhamnoside (2), kaempferol-3-O-beta-D-glucoside (3), kaempferol-3, 7-O-alpha-L-dirhamnoside (4), quercetin-3-O-beta-D-glucoside (5), kaempferol-3-O-beta-D-rutinoside (6).

CONCLUSIONCompouds 1-6 were isolated from this plant for the first time.

Chromatography, Gel ; Dryopteridaceae ; chemistry ; Flavonols ; analysis ; isolation & purification ; Magnetic Resonance Spectroscopy

Dioscin is a natural steroid saponin derived from several plants, showing potent anti-cancer effect against a variety of tumor cell lines. In the present study, we investigated the anti-cancer activity of dioscin against human LNCaP cells, and evaluated the possible mechanism involved in its antineoplastic action. It was found that dioscin (1, 2 and 4 μmol/L) could significantly inhibit the viability of LNCaP cells in a time- and concentration-dependent manner. Flow cytometry revealed that the apoptosis rate was increased after treatment of LNCaP cells with dioscin for 24 h, indicating that apoptosis was an important mechanism by which dioscin inhibited cancer. Western blotting was employed to detect the expression of caspase-3, Bcl-2 and Bax in LNCaP cells. The expression of cleaved caspase-3 was significantly increased, and meanwhile procaspase-3 was markedly decreased. The expression of anti-apoptotic protein Bcl-2 was down-regulated, whereas the pro-apoptotic protein Bax was up-regulated. Moreover, the Bcl-2/Bax ratio was drastically decreased. These results suggested that dioscin possessed potential anti-tumor activity in human LNCaP cells through the apoptosis pathway, which might be associated with caspase-3 and Bcl-2 protein family.
Apoptosis ; drug effects ; Blotting, Western ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Survival ; drug effects ; Diosgenin ; analogs & derivatives ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Enzyme Activation ; drug effects ; Flow Cytometry ; Humans ; Male ; Molecular Structure ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Time Factors ; bcl-2-Associated X Protein ; metabolism

The purpose of this study was to construct a recombinant conditionally replicating adenovirus (CRAd) expressing programmed cell death 5 (pdcd5). Pdcd5 gene was inserted in the E3 region of SG600-a CRAd in which the key genes for virus replication E1a and E1b were controlled under the human telomerase reverse transcriptase promoter (hTERTp) and the hypoxia response element (HRE) respectively, and with a deletion of 24 nucleotides within CR2 region of E1a. The insertion and orientation of all recombined plasmids were confirmed by restriction enzyme digestion and polymerase chain reaction (PCR). The infection efficiencies of a recombined virus carrying enhanced green fluorescent protein (EGFP) in leukemic cell lines were observed by using fluorescence microscope. The relative pdcd5 expression levels of K562 after being infected with SG611-pdcd5 were detected by real-time quantitative PCR. The results showed that the construction of SG611-pdcd5 was completed and confirmed. Pdcd5, hTERTp, HRE, skeleton and fiber11 of recombinant adenovirus SG611-pdcd5 were successfully amplified. The infection efficiencies of SG611-EGFP were all above 70% in both leukemic K562 and MEG-01 cell lines. SG611-pdcd5 expressed pdcd5 with high efficiency in leukemic cells as compared with Ad-pdcd5 or SG611 (p < 0.001). The expression level of pdcd5 increased gradually along with the increase of MOI. It is concluded that the triple-regulated adenovirus of SG611-pdcd5 containing the pro-apopro-tic gene pdcd5 has been successfully established with high pdcd5 expression level in leukemic cells, indicating that the recombinant adenovirus, SG611-pdcd5, promises further development of targeted tumor gene therapy.
Adenoviridae ; genetics ; Apoptosis Regulatory Proteins ; genetics ; Genetic Therapy ; methods ; Neoplasm Proteins ; genetics ; Oncolytic Viruses ; genetics ; Promoter Regions, Genetic ; Telomerase ; genetics

Vascular endothelial growth factor (VEGF) is a specific mitogen for vascular endothelial cells. It has been associated with angiogenesis, growth, metastasis and poor prognosis in solid tumors. Lately, it has been known that VEGF expression is higher in bone marrow from chronic myeloid leukemia (CML) patients than that in normal subjects. However, it is not clarified that the effect of VEGF on the abnormal proliferation of CML cells. In order to explore the effect of autocrine VEGF on CML cells, K562 cells were transfected with the VEGF(121) cDNA sense vector (K562/S) or with the VEGF(121) cDNA antisense vector (K562/As). K562 cells were transfected with the pcDNA(3) vector (K562/V) as the control. Cell proliferation was determined by MTT and colony forming assay in vitro. Flow cytometric Annexin-V-FITC/PI dual labeling technique was performed to observe the effect of VEGF(121) cDNA transfection on apoptosis of K562 cells. Results indicated that K562/S transfectants exhibited a 3-fold increase in VEGF secretion, and K562/As transfectants exhibited a 49% reduction in VEGF secretion. K562/As showed a reduced growth rate and colony forming efficiency as compared to K562/V. K562/S showed an increasing growth rate and colony forming efficiency as compared to K562/V. K562/As had more apoptotic cells than K562/V and K562/S in the same culture condition. These data suggest that VEGF plays an important role in the abnormal proliferation and apoptosis in CML cells through an autocrine mechanism.

OBJECTIVETo evaluate the significance of quantification of WT1 mRNA for monitoring minimal residual disease (MRD) in patients with acute myeloid leukemia (AML).

METHODSWT1 mRNA level was detected with real-time quantitative RT-PCR (RQ-PCR) technique in bone marrow samples from 15 normal subjects (NBM) and 123 AML patients. Sixty-two AML samples were also detected AML1-ETO mRNA expression by RQ-PCR. Simultaneously follow-up of WT1 and AML1-ETO levels were carried out in 50 samples from 8 AML patients. WT1 and AML1-ETO levels were normalized by internal control ABL gene.

RESULTSAll correlation co-efficiencies were over 0.99 for WT1, AML1-ETO and ABL standard curves. Co-efficiencies of both interassay and intraassay variation were below 4%. The WT1 expression levels in NBM were 0.001 to 0.019 with a median level of 0.008. Higher levels of WT1 expression were found in 61 of 67 (91%) newly diagnosed AML patients compared with NBMs and 37 of the 67 (55.2%) showed 100-fold higher WT1 levels than that in NBMs. WT1 mRNA levels were highest in M(4EO) and M(3) and lowest in M(1) and M(5) patients. There was an excellent correlation between WT1 and AML1-ETO gene expression levels (r = 0.88, P < 0.001). WT1 expression levels in three patients who were in continuous complete hematological remission (CHR) were within normal range. In three of four relapsing patients, WT1 expression levels increased 31.4, 11.4 and 4.0 fold respectively one month before hematological relapse.

CONCLUSIONSQuantification of WT1 expression level by RQ-PCR may be used to monitor MRD for most AML patients, but it is less sensitive than fusion gene. Continuous or significant increase of WT1 expression in CHR patients predicts an impending relapse.

Adolescent ; Adult ; Aged ; Bone Marrow ; metabolism ; Child ; Child, Preschool ; Core Binding Factor Alpha 2 Subunit ; genetics ; metabolism ; Female ; Humans ; Leukemia, Myeloid, Acute ; diagnosis ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; metabolism ; pathology ; Oncogene Proteins, Fusion ; genetics ; metabolism ; RNA, Messenger ; genetics ; RUNX1 Translocation Partner 1 Protein ; Recurrence ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; WT1 Proteins ; genetics ; metabolism

OBJECTIVETo compare the immunophenotypic and clinical characteristics between NPM1 mutated acute myeloid leukemia (AML) (NPM1m(+)AML) and unmutated AML(NPM1m(-)AML) not otherwise characterized (NOS) under similar FAB subtypes constituent ratio.

METHODSImmunophenotyping and NPM1 gene mutation type-A, B and D and other leukemic related fusion genes were detected by multiparameter flow cytometry and real time RT-PCR or PCR, respectively. 104 AML patients with NPM1m(+)AML and performed immunophenotyping assay were included, 97 with NPM1m(-)AML.

RESULTSThere were significant difference between the two groups at presentation in terms of sex, white blood count(WBC), platelet counts (PLT), blast ratio, normal karyotype ratio, WT1 expression level, FLT3-ITD mutation positive rate and remission rate of first course of induction therapy (P < 0.05). On the immunophenotype, the expression of early differentiation antigens (CD34, HLA-DR, CD117, CD38), lymphocytic antigens (CD7, CD4, CD19, CD2), myeloid and monocytic differentiation-associated antigens (CD13, CD14, CD15) were lower, and that of CD33 as well as CD123 were higher in NPM1m(+)AML patients. Among them, only CD34, HLA-DR, CD7, and CD4 positive cases were significantly lower in NPM1m(+)AML group than in NPM1m(-)AML group (P < 0.05), the rest of them had significant difference in the number of positive cells (P < 0.05). Above features were further analyzed between the M1/M2 and M4/M5 subgroups. M1/M2 cases retained the women prominent and had a higher WT1 expression level (P < 0.05). The expression of monocytic differentiation-associated antigens including HLA-DR and lymphocytic antigens were higher and that of CD117 were lower in M4/M5 subtype (P < 0.05). Among them, the positive rates of HLA-DR, CD64, CD11b, CD10, CD15, and CD4 were significantly higher in M4/M5 than in M1/M2 in NPM1m(+)AML group (P < 0.05).

CONCLUSIONThe most clinical characteristics in NPM1m(+)AML patients are consistent with reports, but some immunophenotype are different to the previous reports under similar FAB subtypes constituent ratio. The major immunophenotypic features of NPM1m(+)AML patients are lower expression of progenitor, myeloid and lymphoid lineage antigens. Monocytic differentiation-associated antigens are only higher expression in M4/M5 cases when comparison with M1/M2 cases within NPM1m(+)AML group.

Adolescent ; Adult ; Aged ; Antigens, CD ; metabolism ; Child ; Child, Preschool ; Female ; HLA-DR Antigens ; immunology ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; immunology ; Male ; Middle Aged ; Mutation ; Nuclear Proteins ; genetics ; Young Adult