Adult ; Female ; Humans ; Kidney Glomerulus ; metabolism ; pathology ; Lipoproteins ; metabolism ; Male ; Nephrosis, Lipoid ; metabolism ; pathology

To elucidate the role of fetal bone marrow stromal cells (FBMSC) in cooperation with exogenous cytokines in supporting the in vitro expansion of cord blood CD34(+) cells which were purified by negative immunomagnetic selection, FBMSCs were cultured with different combinations of cytokines including SCF, IL-3, IL-6, FL, G-CSF and EPO in a 14-day liquid culture system. The results showed FBMSC plus SCF, IL-3, IL-6, FL and EPO was the most effective combination which increased total nucleated cells, CFU-GM, BFU-E and CD34(+) cells by (692.4 +/- 52.7) fold, (237.1 +/- 106.6) fold, (114.8 +/- 32.8) fold and (25.3 +/- 10.1) fold, respectively. Our studies indicated that fetal bone marrow stromal cells combined with above-mentioned cytokines can efficiently expand cord blood CD34(+) cells.

Objective： To elucidate the role of bone marrow stromal cells in cooperation with exogenous cytokines in hematopoiesis. Methods： Fetal bone marrow stromal cells (FBMSC) was combined with cytokines including SCF,IL-3,IL-6,GM-CSF in a 5-day liquid culture system of adult bone marrow mononuclear cells, then we cultured bone marrow derived CD34+-enriched cells with FBMSC+SCF+IL-3+IL-6+G-CSF+EPO for 2 weeks. Results：FBMSC were in good cooperation with above mentioned exogenous cytokines. When CD34+-enriched cells from adult bone marrow were cultured with combinations of FBMSC, SCF, IL-3, IL-6, G-CSF and EPO, total nucleated cells, CFU-GM, BFU-E and CD34+ cells were increased by 119.6±30.9, 54.6±17.4, 25.2±4.4, 11.1±4.2 folds, respectively. Conclusion：FBMSC in cooperation with exogenous cytokines support the in vitro expansion of human hematopoietic progenitor cells efficiently.

The inhibition activity of 36 flavonoids against CYP1A2 was determined by our previously developed in vitro method. The Comparative Molecular Similarity Indexes Analysis (CoMSJA) approach was used to probe the quantitative relationships between the flavonoids' molecular structural descriptors and their inhibitory activities. A reliable CoMSIA model with the combined electrostatic and hydrophobic fields was derived with the regression coefficient R2 of 0.948 and the cross-validation regression coefficient q2 of 0.630, separately, which is capable of elucidating the quantitative relationship between the 3D structural descriptors of the flavones and their bioactivities. Comparing with flavone, the larger pi-pi conjugated system of alpha-naphthoflavone significantly improved the biologically inhibitory ability. Based on the core structure of the latter, either electropositive substituents or hydrophobic groups at the 6, 3', and 4' ring positions or electronegative counterparts at the 5 ring position, can enhance the inhibitory potency against CYP1 A2 according to the CoMSIA contour maps.
Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 CYP1A2 Inhibitors ; Flavonoids ; chemistry ; pharmacology ; Humans ; Microsomes, Liver ; metabolism ; Models, Molecular ; Molecular Structure ; Quantitative Structure-Activity Relationship

Cardiac Catheterization ; Heart Septal Defects, Ventricular ; therapy ; Humans ; Prosthesis Implantation ; instrumentation ; Septal Occluder Device

OBJECTIVELeukemia is a major cause of death of children in China, which accounts for 50 % of all cancers of children. Data from Tianjin Cancer Hospital was analyzed for mortality of leukemia in children under 20 years from 1981 to 2000 in the city of Tianjin.

METHODSAll physicians and medical staff of the hospitals and clinics in the registry area were responsible for filling out the report forms for every new case diagnosed as malignant tumors. Death certificates for malignant tumors have been registered at the local police station and the residential files were checked. All cancer cases with insufficient information were traced to his/her family and relevant persons worked in the clinic. Tianjin Cancer Registry Center periodically conducted an active re-checking program to review all patient records on cancers that was not registered in this period. Tumors diagnosed in this study were coded according to the International Classification of Diseases for Oncology (ICD-O). Mortality rates were calculated by age, sex and date of death.

RESULTSThe types of acute lymphoid leukemia, acute myeloid leukemia and chronic myeloid leukemia were the most common types of childhood leukemia in Tianjin, comprised 69.3%, 20.9 % and 8.0%, respectively. The mortality for childhood leukemia decreased slowly during the period of 1981 to 2000 in Tianjin. Mortality and morbidity ratios were 0.51.

CONCLUSIONCombined with characteristics of individual forms of childhood leukemia mortality, further epidemiological research is needed to prevent childhood leukemia.

Adolescent ; Cause of Death ; Child ; Child, Preschool ; China ; epidemiology ; Humans ; Infant ; Leukemia ; mortality ; Young Adult

OBJECTIVETo evaluate the clinical efficacy of syndrome differentiation-based treatment with traditional Chinese medicine (TCM) versus losartan therapy in addition to basic treatment for management of proteinuria in patients with chronic kidney disease.

METHODSThis multicenter, randomized, and case-controlled clinical trial was conducted among 81 consecutive patients meeting the inclusion criteria. The patients were randomized consecutively to receive TCM treatments according to the syndrome patterns in TCM (spleen and kidney Qi and Yin deficiency, and spleen and kidney Qi and Yang deficiency, n=60) or oral losartan therapy (50 mg/day, n=21) in addition to the basic treatments. All the patients were followed up for 24 weeks to observe the clinical effects.

RESULTSThe patients in TCM group showed a significantly higher overall response rate (93.33%) than those in losartan group (76.20%, P<0.05). The TCM score in the two groups were all decreased at week 24 as compared with baseline (P<0.01 or P<0.05). The TCM scores in both groups decreased significantly after the treatments as compared with the baseline scores (P<0.05). After a 8-week-long treatment, Scr, eGFR and Cys-C, U-Pro/24 h, and MA/Cr all decreased significantly in TCM group (P<0.05) but showed no significant changes in losartan group (P>0.05).

CONCLUSIONSyndrome differentiation-based TCM treatment in addition to basic treatments can produce satisfactory therapeutic effects on proteinuria in patients with chronic kidney disease by improving the clinical symptoms, reducing TCM symptom scores and proteinuria, and protecting the renal functions.

Adult ; Aged ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Losartan ; therapeutic use ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Phytotherapy ; Prospective Studies ; Proteinuria ; drug therapy ; etiology ; Renal Insufficiency, Chronic ; complications ; drug therapy

OBJECTIVETo investigate whether or not NADPH oxidase (NOX) participates in leptin-induced reactive oxygen species (ROS) production in hepatic stellate cells (HSC) and to explore the possible mechanism.

METHODSHSC-T6 cells (rat hepatic stellate cells line) were divided into nine groups: Group1: leptin (100 ng/ml) treated; Group2-6: leptin treated together with inhibitors that block different ROS-producing systems: diphenylene-iodonium (DPI) (20 micromol/L), Rotenone (20 micromol/L), Metyrapone (250 micromol/L), Allopurinol (100 micromol/L) and Indomethacin(100 micromol/L); Group7: leptin treated together with Janus kinase (JAK) inhibitor AG490 50 micromol/L; Group8: normal control group (treated DMEM with 0.1% DMSO); Group9: negative control group (untreated). Intracellular ROS levels were measured with dichlorodihydrofluorescein diacetate (DCFH-DA) dye assay by Fluorescence microscope and/or flow cytometry. NOX activity was analyzed by using spectrophotometer to calculate the absorbance of NADPH. The mRNA levels of Rac1 and p22Phox were evaluated by RT-PCR.

RESULTS(1) Leptin increased significantly the ROS production as compared to normal control group (92.91+/-4.19 vs.27.56+/-6.27, P<0.01) in HSC-T6 cells. Both the NADPH oxidase inhibitor DPI and AG490 (50 micromol/L) blocked the ROS production, inhibitors of other ROS producing systems had no significant effect on ROS production induced by lepin (P is more than 0.05). (2) Leptin treated HSC-T6 cells for 1 hour up-regulated the NOX activity significantly compared with that in normal control group [(1.90+/-0.22) pmol.min(-1).mg(-1) vs. (0.76+/-0.06) pmol.min(-1).mg(-1), P<0.05]. Furthermore, the NOX activity increased after being treated with leptin for 12 hours and 24 hours than being treated for 1 hour. Leptin-induced up-regulation of NOX activity was inhibited by pretreatment with DPI or AG490. (3) The RT-PCR results indicated that mRNA expressions of Rac1 and p22Phox in HSC-T6 cells with 12 hours of leptin stimulation increased significantly as compared with normal control group (0.41+/-0.13 vs 0.14+/-0.08, 0.45+/-0.12 vs 0.20+/-0.08, all P<0.05), while the DPI and AG490 had no effect on the mRNA expressions of Rac1 and p22Phox.

CONCLUSIONNOX is the main cellular source of the reactive oxygen species (ROS) generated by HSCs in response to leptin stimulation. The mechanism is probably that leptin can directly activate NOX through JAK signal transduction and hence induce the expression of NOX subunit to promote the activity of NOX which generates considerable ROS in HSC.

Animals ; Cells, Cultured ; Hepatic Stellate Cells ; drug effects ; metabolism ; Leptin ; pharmacology ; NADPH Oxidases ; genetics ; metabolism ; Rats ; Reactive Oxygen Species ; metabolism

OBJECTIVETo investigate the clinicopathologic and immunophenotypic features of primary breast lymphoma (PBL) and to discuss the diagnosis of the tumor.

METHODSTwenty-one cases of PBL with follow up data were clinically reviewed. The histologic diagnosis of the tumor was based on the updated WHO Classification of tumors of hematopoietic and lymphoid tissues (2008). Immunohistochemistry was performed by SP method and antibodies selected were as follows: CD20, CD3epsilon, CD10, Bcl-6, MUM-1, CD5, Bcl-2, CD23, CD10, cyclin D1, CD43 and Ki67.

RESULTS(1) All 21 patients were female and the median age of patients was 48 years. The right and the left breasts were involved in 11 (52.4%) and 10 patients (47.6%), respectively. According to Ann Arbor staging system, 20 cases were stage I-II (95.2%), and the remaining case was stage IV (4.8%). For the international prognostic index (IPI), 19 cases were score 0-1, and 2 cases were score 2-3. For ECOG score, 19 cases were 0, and the remaining 2 cases were 1. (2) Histologically, all 21 cases (100%) were DLBCL. Immunohistochemically, the frequency of antigen expression was as follows: CD20 (100%), MUM-1 (14/21, 66.7%), bcl-6 (5/21, 23.8%), CD10 (0), bcl-2 (13/21, 61.9%), CD5 (1/21, 4.8%); Ki-67 index: 10 cases (47.6%) were less than 59%, with the expression of seven cases (33.3%) being 60% - 89%, and more than 90% in the remaining four cases (19.1%). The median Ki-67 index was 60%. All the cases were considered non germinal center B-cell-like type of DLBCL. (3) Follow-up data was available in 64% of the cases. One, two and five-year survival rates were 11 cases, 7 cases and 3 cases, respectively.

CONCLUSIONAll the cases of PBL in the current study were DLBCL, non germinal center B-cell-like type, and a diagnosis of PBL can only be established after excluding breast involvement in systemic lymphoma.

Adult ; Aged ; Antigens, CD20 ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Interferon Regulatory Factors ; metabolism ; Ki-67 Antigen ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; metabolism ; pathology ; surgery ; Mastectomy, Radical ; Middle Aged ; Neoplasm Staging ; Prednisone ; therapeutic use ; Proto-Oncogene Proteins c-bcl-6 ; metabolism ; Retrospective Studies ; Survival Rate ; Vincristine ; therapeutic use ; Young Adult

To further explore the mechanism of congenital pyrimidine 5'-nuleotidase I (P5'N-I) deficiency, on the basis of purification of the protein, the molecular weight and amino acid composition were analysed by mass-spectrograph and amino-acid analyzer, microsequencing and bioinformation analysis of P5'N-I were performed after it was hydrolysed by trypsin. The results showed that three fractions were found in the purified P5'N-I and their molecular weights were 26,952.9, 55,476 and 110,938, respectively. The sequence from one of the peptide fragments was I-E-G-P-T-I-R-Q-I-E. The homologous sequence was not found after comparision with the ten-amino-acid sequence in GenBank by blast procedure. Amino acid analysis indicated that P5'N-I was composed of 18 amino acids at least, and 243 amino acid residues. In conclusion, the enzyme might be an allosteric enzyme, there might be homologous dimer or tetramer in physiological status of normal human erythrocyte, the microsequence could be designed as the probe for fishing the genes of interest. The composition of amino acid might be an important information in determination of its protein primary structure.
5'-Nucleotidase ; blood ; chemistry ; isolation & purification ; Amino Acid Sequence ; Amino Acids ; analysis ; Chromatography, High Pressure Liquid ; Electrophoresis, Polyacrylamide Gel ; Erythrocytes ; enzymology ; Humans ; Mass Spectrometry ; Molecular Weight ; Peptide Fragments ; chemistry ; Sequence Analysis, Protein