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

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

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 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

The objective of this study was to estimate a novel apoptosis-promoting molecule PDCD5 expression in the bone marrow cells from adult acute myeloid leukemia (AML) for investigation of its significance in the pathogenesis of AML. Flow cytometry assay was used for detection of PDCD5 expression in the different groups of cells from bone marrow of AML patients and normal controls by using 21 monoclonal antibodies with different fluorescent markers. The PDCD5 expressions in bone marrow cells from some AML patients and normal controls were also detected by Western blot. The results showed that the mean PDCD5 fluorescence intensity in bone marrow nucleated cells (MNC) from the bone marrow of 36 untreated AML patients was significantly lower than that from the bone marrow of 30 normal controls (3059 +/- 1392) vs (7432 +/- 1261) (P < 0.01). The mean PDCD5 fluorescence intensity was lower in the marrow granulocytes, monocytes, blast cells, and lymphocytes from untreated AML patients than that from normal (3939 +/- 2121) vs (8367 +/- 1045); (3156 +/- 1635) vs (5917 +/- 2329); (2824 +/- 1592) vs (3998 +/- 2106); (1474 +/- 816) vs (3355 +/- 2042) respectively, (all P < 0.01). Western blot analysis demonstrated that PDCD5 expression was significantly decreased in the AML cells, as compared with normal cells. It is concluded that PDCD5 expression in MNC in untreated AML patients is lower than that in the normal. PDCD5 expression in the marrow granulocytes, monocytes, blast cells, and lymphocytes of untreated AML patients is significantly lower than that in the normal. It suggests that the abnormally low expression of PDCD5 may be involved in the pathogenesis of AML.
Apoptosis ; physiology ; Apoptosis Regulatory Proteins ; metabolism ; Bone Marrow Cells ; metabolism ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; pathology ; Neoplasm Proteins ; metabolism

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 explore the specificity of anti-phosphotyrosine monoclonal antibody PY20 in bcr-abl+ cells and its possible clinical applications.

METHODSBcr-abl cell lines( K562, MEG-01) and bcr-abl- cells lines( Jurkat, MCF-7 )were stained with PY20. Phosphotyrosine protein of K562 and MEG-01 cells was detected by flow cytometry before and after treatment with imatinib. Phosphotyrosine protein in bone marrow cells from 49 patients with chronic myeloid leukemia (CML), Ph+ acute lymphoblastic leukemia(Ph(+) -ALL), Ph- ALL, acute myeloid leukemia (AML-M1, M2, M3, M5, FAB classification), chronic lymphocytic leukemia (CML) and 3 normal donor. Positive cells over 5% of total cells was considered positive cases for phosphotyrosine protein. The level of tyrosine phosphorylation was determined by median fluorescence intensity (MFI).

RESULTSBcr-abl cell lines and marrow cells from 10 CML patients and 8 ALL patients were all PY20-positive, while bcr-abl- cell lines and marrow cells from 18 leukemia patients and 3 normal donor were all PY20-negative. MFI decreased remarkably after blocked by imatinib in K562 cells and MEG-01 cells. The positive cell percent of marrow cells from 10 newly diagnosed CML patients and 9 imatinib-sensitive CML patients was (54.20 +/- 19.82)% and (14.84 +/- 6.17)% (P < 0.05), while that of 2 cases of imatinib-resistant was 64.3% and 57.2%. There was significant difference of MFI between imatinib-resistant patients and imatinib-sensitive patients (99.42 +/- 4.87 vs 46.41 +/- 4.67, P < 0.01).

CONCLUSIONPY20 monoclonal antibody is highly specific for bcr-abl+ cells. It might be useful in rapid detection of bcr-abl+ cells and sensitivity to imatinib of CML patients.

Antibodies, Monoclonal ; analysis ; Bone Marrow Cells ; metabolism ; Flow Cytometry ; Fusion Proteins, bcr-abl ; analysis ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; Leukemia, Myeloid, Acute ; metabolism ; Phosphotyrosine ; analysis ; immunology

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.

In order to investigate the features of M-bcr/abl and m-bcr/abl fusion transcripts in patients with chronic myeloid leukemia (CML) after allogeneic stem cell transplantation (SCT), M-bcr/abl and m-bcr/abl fusion transcripts were sequentially detected by RT-PCR technique in 72 CML patients after SCT. The results showed that M-bcr/abl positive rate (79.2%, 42/53) within 6 months after SCT was remarkably higher than that in 6-12 months group (34.3%, 11/32) and >or= 12 months group (35.1%, 13/37) (P < 0.001), and the clinical relapse rates in corresponding periods were 1.9% (1/53), 0% (0/32) and 16.2% (6/37) respectively. M-bcr/abl and m-bcr/abl fusion transcripts occurred in 5 of 6 clinically relapsed patients. In period of more than 6 months after transplantation, none of 17 M-bcr/abl(+) samples from 14 patients in cytogenetic remission appeared positive reaction of m-bcr/abl. It is concluded that M-bcr/abl(+) fusion transcript still existed in most patients after SCT, and usually disappeared within 6 months. Existence of M-bcr/abl is not a clinical relapse marker in CML patients. Simultaneous detection of M-bcr/abl and m-bcr/abl fusion transcripts can be helpful for monitoring residual disease.
Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Fusion Proteins, bcr-abl ; genetics ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; therapy ; Male ; Middle Aged ; RNA, Messenger ; analysis ; Recurrence ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Homologous