AIM: To study compatibility rationality of combination of Paeonia lacliflora and Glycyrrhiza uralensis. METHODS: The effective combination of paeoniflorin(44% purity),glycyrrhizic acid(50% purity) and liquorice flavones(52% purity),glycyrrhizic acid(50% purity) and liquorice flavones(52% purity) were respectively administered to rats.Pharmacokinetic change of these constituents in rat blood was studied. RESULTS: The pharmacokinetic parameters of these constituents in rat blood showed that the increases in AUC and C_(max) of effective combination group were more than that of glycyrrhizic acid group or that of liquorice flavones group.T_(max) of the former was extended with respect to the latters.Clearance of effective combination markedly slowed down. CONCLUSION: The effective combination of paeonia lacliflora and Glycyrrhiza uralensis have the advantage of either Paeonia lacliflora or Glycyrrhiza uralensis.

Objective To investigate the changes of CD4+ CD25+ regulatory T lymphocyte (Treg) and expressions of folkhead helix transcription factor 3 (FoxP3) in intestinal mucosa in human immunodeficiency virus (HIV) infected patients. Methods Twenty-one HIV infected patients and 17 control subjects without HIV infection were included in this study. The expression of FoxP3, which was considered as a specific marker of CD4+ CD25 + Treg, was detected in intestinal mucosa specimens from HIV infected patients by immunohistochemistry. Meanwhile, the in situ expression of CD4+ T lymphocyte was also determined by immunohistochemistry. The data were analyzed by t test. Results The positive labeling index of CD4+ T lymphocyte in intestinal mucosa was significantly lower in HIV infected patients compared to the controls (11. 56%±4. 44% vs 43. 49% ±8. 90% ,t=-11. 86,P<0. 01). The positive labeling index of FoxP3 in intestinal mucosa was also significantly lower in HIV infected patients compared to the controls (0.46% ± 0.20% vs 1. 18% ± 0. 44% ,t= - 5. 98,P<0.01). Conclusion The depletion of CD4+ CD25+ Treg is accompanied with the depletion of CD4 + T lymphocyte and the reduction of FoxP3 expression in intestinal mucosa of HIV infected patients.

AIMTo provide experimental basis for clinical use of cryopreserved dendritic cells (DCs) by investigating the biological properties of cryopreserved DCs derived from cord blood.

METHODSThe biological discrepancies between unfrozen DCs (UFDC) derived from unfrozen cord blood (UFCB) and DCs from cryopreserved cord blood (CPCB) or cryopreserved DCs (CPDCs) were explored by morphological observation and immunophenotype analysis. Moreover, the stimulating index (SI) in mixed lymphocyte culture and cytotoxic eliminating rate (ER) were measured by MTT.

RESULTSThe TBR of CPCB and CPDCs were 95.8% and 88.7% respectively. Compared to UFCB, CPCB differentiated toward DCs in a delayed and decreased mode, displayed lower expression of CD1a, CD83 and HLA-DR, and had reduced SI and ER. Similarly, the CPDCs became adherent DCs later and grew less in number than UFDCs. After culture, the expression of CD1a, CD83 and HLA-DR as well as SI and ER were lower in CPDCs than those in UFDCs.

CONCLUSIONThough the biological properties of CPCB and CPDCs were injured after cryopreservation, they still had relatively complete basic function. Their recoveries rate were > 85%.

Cell Proliferation ; Cells, Cultured ; Cryopreservation ; Dendritic Cells ; cytology ; Fetal Blood ; cytology ; Humans

AIMTo observe the basical properties of adherent stromal cells in culture derived from cryopreserved bone marrow cells (BMCs), and to provide laboratory evidences for clinical application of cryopreserved stromal cells.

METHODSFresh BMCs and adherent stromal cells cultured for 14 days in Dexter long-term culture system (fresh stromal cells) plus 5% DMSO-6% HES cryopreservatives were frozen in -80 degrees C refrigerator, and cryopreserved in - 196 degrees C liquid nitrogen for 2 weeks (the former is called cryopreserved BMCs, the latter called cryopreserved stromal cells). These cells were cultured in Dexter long-term culture system after they were thawed. We have examined the growth features, constituents and stimulating functions of the culture of these cells.

RESULTSGrowth features: Cryopreserved BMCs produced adherent stromal cells, cell clusters and cell layer 1-2 days later than fresh BMCs. Cryopreserved stromal cells formed cell layer in 2nd day of culture, and were 12-18 h later than fresh stromal cells. Cryopreserved BMCs and stromal cells proliferated significantly less than fresh BMCs and stromal cells. Constituents: The ratio of fibrocytes and endothelial cells were lower, and the ratio of macrophages and fat cells were higher in culture of cryopreserved BMCs than that in fresh BMCs. The measurements in culture of cryopreserved stromal cells were much more significant compared with that in fresh stromal cells. Numbers of cells containing apoptic bodies in culture of cryopreserved BMCs and stromal cells were more than that in fresh BMCs and stromal cells. TBRR of cryopreserved BMCs and stromal cells were 92.5% and 89.5% respectively. The expression rates of CD14 and HLA-DR in culture of cryopreserved BMCs and stromal cells were higher than that in fresh BMCs and stromal cells, and just in contrast with the expression rates of CD45 and CD33. Stimulating functions: CAFA and LTC-IC on the stromal cell layer derived from cryopreserved BMCs, cryopreserved stromal cells, fresh BMCs and fresh stromal cells were all growing well, and there were no significant differences among these groups.

CONCLUSIONBiological properties of adherent stromal cells derived from BMCs and stromal cells were injured slightly and still maintained completely after cryopreservation with 5% DMSO-6% HES cryopreservatives.

Bone Marrow Cells ; cytology ; Cell Survival ; Cells, Cultured ; Cryopreservation ; Humans ; Stromal Cells ; cytology

The aim was to investigate the cytolytic activity of cytotoxic T lymphocytes (CTL) induced by dendritic cells (DC) derived from human cord blood in vitro. Human cord blood mononuclear cells (CBMNC) were cultured in vitro with addition of various cytokines. DC was confirmed by morphology, immune phenotype and capacity of stimulating MLR (mixed lymphocyte reaction). CTL were generated through the co-culture of autologous T lymphocytes and DC. (51)Cr-release assays were performed for the measurement of cytotoxicity of CTL. The results showed that distribution of the subgroups of T lymphocytes in CBMNC was similar to that in adult peripheral blood. The percentage of CD1a-expressing cells in CBMNC was very low, merely (0.41 +/- 0.09)%. During culture, DC became larger and more irregular in shape. Spiny dendrites or multiple cell processes in morphology emerged on the surface of DC. Among the cell populations at 15 days of culture, there were (28.4 +/- 3.55)% of CD1a-expressing cells, (63.67 +/- 23.33)% of CD86-positive, (8.7 +/- 1.49)% of CD83-positive and (32.5 +/- 1.53)% of HLA-DR-positive cells. DC derived from CBMNC is capable of stimulating the proliferation of allogeneic lymphocytes in MLR. CTL derived from autologous T lymphocytes induced by dendritic cells pulsed with lysates of HL-60 cells, possessed specific cytolytic effects against HL-60 cells. In conclusions, relatively high percentage of CD1a-expressing cells can be generated in culture system of this study. DC derived from cord blood is able to induce the production of anti-leukemic CTL in vitro.
Antigens, CD1 ; analysis ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Fetal Blood ; immunology ; Humans ; Immunophenotyping ; Leukemia ; immunology ; T-Lymphocytes, Cytotoxic ; immunology

Preparative HPLC was used to prepare ferulic acid, senkyunolide I and senkyunolide H from Ligusticum chuanxiong. The separation was conducted on a Shim-Pack Prep-ODS (20.0 mm x 250 mm, 5 microm) column with the mobile phase of methanol-0.2% glacial acetic acid (50:50)at the flow rate of 5 mL x min(-1). The detection wavelength was 278 nm, and the purity of each compound was detected by HPLC analysis. Spectral data analyses including UV, ESI-MS and NMR were used to identify their structures. This method is simple, fast, which is suitable for preparation of standard reference of ferulic acid, senkyunolide I and senkyunolide H from L. chuanxiong and can meet the requirement of new drug research and development.
Benzofurans ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; chemistry ; isolation & purification ; Ligusticum ; chemistry

OBJECTIVETo study the spasmolysisy activity constituents of the Radix Paeoniae alba and Radix et Rhizoma compound.

METHODThe three effective compounds of Radix Paeoniae and Radix et Rhizoma were arrayed to eight groups by orthogonal design rat, serum samples were collected after administered in these groups, and these serum samples were used to carry out the experiments of serum pharmacology of spasmolysis. The inhibition ratios were assigned as pharmacodynamic variable X, the areas of characteristic peaks on HPLC fingerprints were assigned as variable Y, and the correlation of X and Y was studied in order to research the activity spasmolysisy compounds of effective constituents. Serum samples were analyzed by HPLC-ESI-MS/MS.

RESULTThe correlation of fingerprints and pharmacology showed that four compounds were the main spasmolysisy activity compounds. The correlation parameter was greater than >0.4, whose value were 0.774, 0.678, 0.566 and 0.472 respectively, and their retention times were 8.03, 51.7, 54.59 and 72.43 min respectively. They were metabolites of effective compounds.

CONCLUSIONThis study makes a valid approach for understanding the material foundation of TCM by establishing the correlation of fingerprints and pharmacology of activity compounds in vivo.

Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; pharmacology ; Glycyrrhiza uralensis ; chemistry ; Male ; Muscle Contraction ; drug effects ; Paeonia ; chemistry ; Rats ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

This study was aimed to explore the expression of DNA-dependent protein kinase catalytic subunit (DNA-PKcs) in adult acute leukemia and its correlation with clinical characteristics, karyotype and prognosis. Indirect immunofluorescent cytometry was used to detect the expression of DNA-PKcs in bone marrow mononuclear cells of 105 patients with acute leukemia before chemotherapy and 41 of them after 2 cycles of chemotherapy. Cytogenetic data were obtained from 26 of them by R band karyotypic analysis. The results showed that the expression of DNA-PKcs was correlated with higher WBC count level in peripheral blood (p < 0.05), but was not obviously associated with median age, gender, percentage of bone marrow blasts, clinical classification, median hemoglobin level and median platelet count (p > 0.05). The middle and strong positive expression of DNA-Pkcs in non-remission group was significantly higher than that in remission group (p < 0.05). The positive rate of DNA-PKcs in abnormal chromosome group was significantly higher than that in chromosome normal group (p < 0.05). It is concluded that the DNA-PKcs expression level is closely related with the increased WBC count, and the expression of DNA-PKcs is correlated also with karyotype and clinical prognosis in adult acute leukemia.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Chromosome Aberrations ; DNA-Activated Protein Kinase ; genetics ; metabolism ; Female ; Humans ; Karyotype ; Leukemia ; diagnosis ; genetics ; metabolism ; Male ; Middle Aged ; Nuclear Proteins ; genetics ; metabolism ; Prognosis ; Young Adult

he activity of MMP-2 and MMP-9 via ERK1/2 signaling pathway.

During the past two decades tremendous efforts have been made by the medical community, especially in the fields of forensic medicine and pediatrics, to better understand the etiology, epidemiology and pathophysiology of SIDS. There have been many SIDS reports from developed countries, but few from developing Asian countries. Despite a recent significant decrease in the incidence of SIDS in many developed countries, SIDS continues to be the most common cause of post-neonatal infant death in these countries. This article analyzes the SIDS data (1990-2006) from the Office of the Chief Medical Examiner for the State of Maryland, USA, along with review of the literature with regard to the history, epidemiological and pathophysiological characteristics of SIDS, as well as the recent advances in SIDS research. The changing trends in the diagnosis of SIDS and current challenges to its forensic investigation are also discussed.
Arrhythmias, Cardiac/complications* ; Forensic Medicine ; Humans ; Infant ; Risk Factors ; Sleep Apnea Syndromes/complications* ; Sudden Infant Death/etiology* ; United States/epidemiology*