In order to study the accumulation of Fritillaria thunbergii cultivar, peimine content in Xiaye, Kuanye, Duozi and Xiaosanzi bulbs of different sizes and parts was determined by HPLC-ELSE. The results indicated that the peimine content varied significantly with the cultivar type, the size and part of bulb. The distribution laws of peimine were as follow: Xiaosanzi > Duozi > Xiaye > Kuanye, small-size bulb > big-size bulb, core bud > scale. The peimine yield per plant in Duozi was the highest.
Cevanes ; analysis ; Chromatography, High Pressure Liquid ; Fritillaria ; chemistry ; growth & development

Objective To study the MRI manitestation of juvemle acute pure cartilage fracture of the knee joint.Methods The MRI changes of cartilage,subcartilage low signal line and subcartilage bone were analysed retrospectively in 26 juvenile patients with acute pure cartilage fracture confirmed by arthroscopy.Sagittal and coronal MRI scanning were performed in 26 patients.Using fast low angle shot fat saturation T_1-weighted image(FLASH-FS-T_1WI)sequences,spin echo T_1-weighted image(SE-T_1WI)and fast imaging with steady-state precession three dimensional fat saturation T_2-weighted image(FISP-3D-FS- T_2WI)sequences in sagittal plane,SE-T_1WI and multi echo data image combination T_2-weighted imaging (MEDIC or ME-T_2WI)in coronal plane.Using ME-T_2WI sequence,axial plane MRI scanning in 5 patients.Results Twenty-seven sites of 26 patients include 8 patella,7 femoral medial condyle, 11 femoral lateral condyle and 1 tibial plateau.Three types pure cartilage fracture were observed,totally defect of the cartilage in 7 sites(include 3 patella,2 femoral medial condyle,1 femoral lateral condyle and 1 tibial plateau),fissuring fracture in 3 sites(include 2 femoral medial and 1 femoral lateral condyles), superficial defect of the cartilage in 17 sites(include 5 patella,3 femoral medial and 9 femoral lateral condyle).Corpus liberum was found in 21 patients'knee joints by arthroscopy,but only 3 cases by MRI. Bone bruise was detected,and subcartilage low signal lines were normal.Conclusion Using FLASH-FS- T_1WI,SE-T_1WI,FISP-3D-FS-T_2WI and ME-T_2WI sequences,sagittal and coronal MRI scanning in femoral and tibial plateau pure cartilage fractures,and using ME-T_2WI sequence axial scanning in patella r cartilage fractures may show the position,extension and types of the acute pure cartilage fracture of the knee joint. MRI is the hest non-invasive method for studying cartilage fracture.

Objective To observe the effects of 2 Gy γ-ray irradiation on regulatory T cells (Tregs) and Th17 cells and immune balance of mice.Methods A total of fifty C57BL/6 male mice were randomly divided into two groups,the irradiated group exposed to 2 Gy of whole body γ-ray irradiation,and the control group sham-irradiated.At 1,3,7,14 and 28 d after radiation,changes of peripheral haemogram were detected respectively and Tregs in peripheral blood,thymus and spleen and Th17 cells in spleen were analyzed by flow cytometry.Results Compared with control group,the number of peripheral blood white cells (WBC) and lymphocyte in irradiated group reduced significantly post-irradiation (t =8.89-33.54,P ＜ 0.05),while the cell number of peripheral CD4 + CD25 + Tregs post-irradiation rose but not significantly.Thymic Treg cells increased 1 and 3 d post-irradiation(t =-6.45,-10.59,P ＜0.05),but reduced 28 d post-irradiation (t =5.34,P ＜ 0.05).Splenic Treg cells ascended obviously from 1 to 14 d post-irradiation (t =-6.82-3.89,P ＜ 0.05).After irradiation splenic Th17 cells increased at 1 d,and reached the maximal level at 3 d (t =-2.42,P ＜ 0.05),more obviously than splenic Treg cells.The reduction of Treg/Th17 ratio from 1 to 14 d post-irradiation disturbed Treg/Th17 balance and made it drift to the direction of Th17 (t =4.02-8.04,P ＜ 0.05).Conclusions Treg/Th17 imbalance plays an important role in immune injury induced by irradiation.

Cell Transformation, Neoplastic ; drug effects ; genetics ; Dimethyl Sulfoxide ; pharmacology ; Electroporation ; methods ; HL-60 Cells ; Humans ; Membrane Proteins ; genetics ; Neoplasm Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Stromal Interaction Molecule 1 ; Transfection

Objective - To optimize the extraction and quantification methods for the determination of S phenylmercapturic acid - Methods (SPMA) in urine with performance liquid chromatography mass spectrometry. The urine was hydrolyzed with 50.0% sulfuric acid. The hydrolysate was purified by solid phase extraction column. Purified samples were separated by C18 chromatographic column and detected by tandem mass spectrometry. The isotope labeled SPMA was used as the internal Results - standard. The internal standard curve was used for quantification. The linear range of SPMA was 0.50 50.00 μg/L with the correlation coefficient of 0.999 8. The detection limit and the lower limit of quantification were 0.05 and 0.17 μg/L, - - - - respectively. The recovery rate was 97.0% 102.0%. The within run and between run relative standard deviation were 0.6% 1.0% - and 1.7% 6.5%, respectively. The mass concentration of urinary SPMA in the occupational benzene exposure group was - vs P higher than the non occupational benzene exposure group by this method (median: 2.81 0.28 μg/g creatinine, <0.05). Conclusion Compared to the national standard method, this optimized method of solid phase extraction and internal standard for quantification eliminates the matrix effect. This method is accurate and precise, and is suitable for the determination of SPMA acid in urine.

Objective To explore the relationship between the Interleukin 6 , matrix metalloproteinases 2 and early embryo arrest. Methods Real time-PCR was used to measure the mRNA expression of IL-6 and MMP-2 and immunohistochemistry (IHC, SP method)was used to measure the location and expression of the two different kinds of protein in villus. ELISA was used to measure the level of IL-6 in serum. Results Real-time PCR and IHC showed that the expression levels of IL-6 was significantly lower in experimental group than in control group, and MMP-2 was significantly higher than the control group (P < 0.05). Differenc of IL-6 level in serum between the two groups was not statistically significant (P > 0.05). Conclusion Proper expressions of IL-6 and MMP-2 in the villus tissue play a key role in the maintenance of early pregnancy.

OBJECTIVETo study whether aristololactam I (AL-I) induces injury in human renal proximal tubular epithelial cells.

METHODCultured human renal proximal tubular epithelial cell line HK-2 was used as the subject. Aristolochic Acid I (AA-I) was used as a positive control. Cell toxicity of AL-I was detected by LDH releasing rate. Cell apoptosis was evaluated by cellular morphology, DNA content and expression of cell membrane phosphatidylserine (PS). The secretion level of fibronectin (FN) and TGF-beta1 in HK-2 cells were assayed by ELISA.

RESULTAL-I had a direct toxicity on HK-2 in a dose dependent manner from 2.5 mg x mL(-1) to 20 mg x mL(-1); In these range of concentration, AL-I could induce cell apoptosis which was detectable by measurements of morphology, DNA content and expression of PS. AL-I could stimulate the secretion of FN and TGF-beta1. The potency of AL-I cell toxicity was higher than AA-I at the same concentration. The effects of AL-I on apoptosis, secretion of FN and TGF-beta1 were all weaker than AA-I.

CONCLUSIONAL-I as one metabolite of AA-I in vivo induces direct injury in renal proximal tubular cells. Its effects are similar to those of AA-I. AL-I may be one of toxic metabolites in Chinese herbs containing AA which participate in renal damage and fibrosis.

Apoptosis ; drug effects ; Aristolochia ; chemistry ; Aristolochic Acids ; administration & dosage ; isolation & purification ; toxicity ; Cell Line ; Cell Membrane ; metabolism ; DNA ; genetics ; Diploidy ; Dose-Response Relationship, Drug ; Epithelial Cells ; drug effects ; Humans ; Kidney Tubules, Proximal ; cytology ; metabolism ; Plants, Medicinal ; chemistry

This study was aimed to explore the expression and significance of DNMT1 gene in bone marrow of patients with acute myelogenous leukemia (AML). The expression of DNMT1 gene was detected by real-time PCR in 30 healthy people and 126 AML patients. The results showed that the expression level of DNMT1 gene was lower in the 30 healthy people and was higher in AML patients. There was a marked decline in the expression level of DNMT1 gene after complete remission (CR) as compared with the initial treatment. The expression level of DNMT1 gene did not correlated with age, sex and the clinical characteristics at initial diagnosis such as white blood cell count and chromosomal karyotype in AML patients. The CR rate in AML patients with low expression level of DNMT1 gene was lower than that in those with high expression level. It is concluded that bone marrow DNMT1 gene level may play an important role in AML pathogenesis and can serve as an index in evaluating AML prognosis.
Adolescent ; Adult ; Aged ; Bone Marrow ; metabolism ; pathology ; Case-Control Studies ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; metabolism ; Female ; Humans ; Karyotyping ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Prognosis ; Young Adult

OBJECTIVETo explore the effects of beta-elemene combined with aclarubicin on the induction of HL-60 cell apoptosis and its mechanisms in antileukemia therapy.

METHODSHL-60 cells were treated for 20 hours with different dose of aclarubicin (0.05, 0.10, 0.25 microg/ml) or with different concentrations of beta-elemene (10, 20, 40 microg/ml) in the presence or absence of aclarubicin (0.10 microg/m). The apoptotic rate was analyzed by flow cytometry (FCM), the productions of PGE2 in culture supernatants was detected by competitive ELISA and the expressions of COX-2 and NF-kappaB activity in HL-60 cells by Western blot.

RESULTSLower concentration of aclarubicin (0.05, 0.10 microg/ml) didn't affect apoptotic rate, and COX-2, NF-kappa B and PGE2 expression on HL-60 cells. Combined treatment of beta-elemene and aclarubicin (0.10 microg/ml) enhanced the apoptotic effect and down-regulated COX-2, NF-kappaB and PGE2 expressions. There was a positive correlation between the effects and beta-elemene concentrations.

CONCLUSIONbeta-elemene enhances aclarubicin-mediated apoptotic effect, down-regulation of COX-2 and their inducing products PGE2 in HL-60 cells by suppressing activitation of NF-kappaB.

Aclarubicin ; Apoptosis ; drug effects ; Cell Line, Tumor ; Down-Regulation ; HL-60 Cells ; Humans ; NF-kappa B ; metabolism