OBJECTIVETo investigate the expression and procoagulant activity of phosphatidylserine (PS) on the normal peripheral blood cells of adults.

METHODSNormal peripheral blood samples were collected from 10 healthy volunteers (5 ml from each volunteer), platelets, neutrophils, lymphocytes and erythrocytes were isolated. The expression and procoagulant activity of PS on normal blood cells were identified by flow cytometry, inhibition test with lactadherin as PS probe and coagulation anticoagulant, respectively.

RESULTSThere was PS expression on a few normal blood cells (9.1%, 5.4%, 3.9% and 3.2% in platelets, neutrophils, lymphocytes and erythrocytes, respectively). The PS on these normal blood cells in vitro showed significant procoagulant activity. The plasma recalcification time was shortened by 47%, 36.5%, 25% and 12.5% by platelets, neutrophils, lymphocytes and erythrocytes, respectively; the formation of factor Xa (through both intrinsic and extrinsic pathways) and thrombin was also increased by 13% - 26% by platelets, neutrophils, lymphocytes and erythrocytes, respectively.

CONCLUSIONThe PS on normal blood cells in vivo may play a crucial role in the coagulation cascade.

Adult ; Blood Cells ; metabolism ; physiology ; Blood Coagulation Tests ; Female ; Flow Cytometry ; Humans ; Male ; Phosphatidylserines ; metabolism

OBJECTIVETo investigate the changes in the activity of nuclear factor-kappaB (NF-kappa B) in mice with dextran sulphate sodium (DSS)-induced rat colitis and its modulalorg effect on intercellular adhesion molecule-1 (ICAM-1) expression.

METHODSTwenty normal male mice were randomized into DSS group and normal saline (NS) control group according to a matched-pair design. From days 1 to 7, the mice in DSS group were subjected to oral administration of 5%DSS solution, and from days 8 to 20, NS was given instead, for a total of 3 cycles. In the control group, only NS was administered. The colonic pathology was observed using HE staining and the mucosa 1 damage was scored for each mouse. The DNA-binding activity of NF-kappa B was tested by electrophoretic mobility shift assay, and the expressions of ICAM-1 and NF-kappa B p65 were detected using immunohistochemistry.

RESULTSThe DNA-binding activity of NF-kappa B was significantly increased in DSS group as compared with NS group. ICAM-1 and p65 expressions were detected in the nuclei of the vascular endothelial and inflammatory cells, especially in the mucosa and submucosa, but such positive cells were seldom observed in NS group. A positive correlation was found between the DNA-binding activity of NF-kappa B and ICAM-1 expression.

CONCLUSIONNF-kappa B activation is an important event in the development of DSS-induced colitis in that activated NF-kappa B upregulates ICAM-1 expression during colonic inflammation.

Animals ; Colitis ; chemically induced ; metabolism ; DNA ; metabolism ; Dextran Sulfate ; Electrophoretic Mobility Shift Assay ; Immunohistochemistry ; Intercellular Adhesion Molecule-1 ; biosynthesis ; Male ; Mice ; Mice, Inbred BALB C ; NF-kappa B ; metabolism ; Protein Binding ; Random Allocation ; Transcription Factor RelA ; metabolism

Bio-active components from Carthamus tinctorius were separated on the basis of antioxidant capacities in vitro. The antioxidant capacity was investigated on the basis of the ability to scavenge DPPH radical, ABTS radical and reduce Fe3+ of different polar fractions. Furthermore, the chemical compounds were isolated from bio-active fraction, and were evaluated for the antioxidative effects. Five major components were isolated and identified from water extract as 6-hydroxykaempferol 3,6,7-tri-O-β-D-glucoside(1), 6-hydroxykaempferol 3-O-β-rutinoside-6-O-β-D-glucoside (2), 6-hydroxykaempferol 3-O-β-D-glucoside (3), hydroxysafflor yellow A (4) and anhydrosafflor yellow B (5). By evaluating and comparing the antioxidative effects of different fractions and obtained compounds, the results showed that water extract displayed significantly high antioxidative activities and 6-hydroxykaempferol glycosides and quinochalcone C-glycosides were found as main contribution for antioxidant property.
Antioxidants ; isolation & purification ; metabolism ; pharmacology ; Benzothiazoles ; metabolism ; Biphenyl Compounds ; metabolism ; Carthamus tinctorius ; chemistry ; Chalcone ; analogs & derivatives ; isolation & purification ; metabolism ; pharmacology ; Ferric Compounds ; metabolism ; Free Radicals ; metabolism ; Kaempferols ; isolation & purification ; metabolism ; pharmacology ; Oxidation-Reduction ; drug effects ; Picrates ; metabolism ; Plant Extracts ; isolation & purification ; metabolism ; pharmacology ; Plants, Medicinal ; chemistry ; Quinones ; isolation & purification ; metabolism ; pharmacology ; Sulfonic Acids ; metabolism ; Water ; chemistry

OBJECTIVETo probe into the clinical effect of needle-pricking therapy for treatment of polycystic ovarial syndrome.

METHODSOne hondred and twenty-one cases of polycystic ovarial syndrome were divided into a needle-pricking therapy group of 61 cases and a medication group of 60 cases with randomized and controlled method. The needle-pricking therapy group were treated by needle-pricking therapy at sacral plexus stimulating points on both sides of the spine and lateral points of Dazhui (CV 14), and the medication group by oral administration of domiphen and intramuscular injection of chorionic gonadotropin (HCG). Levels of hormones and symptoms in the patients before treatment, after treatment of 3 cycles and at the sixth cycle after treatment were investigated.

RESULTSAfter treatment of 3 cycles, the level of hormone and B type ultrasound examination were significantly improved in the two groups (P < 0.01). At the sixth cycle after treatment, the conditions of the patients in the medication group were returned to the original levels before treatment, while the conditions in the needle-pricking therapy group still kept at the post-therapeutic level, and their menstruation and ovulation restored to normal state, and the ovulation mucosa and the pregnancy rate were significantly higher than those in the medication group (all P < 0.01).

CONCLUSIONNeedle-pricking therapy has obvious effect on polycystic ovarial syndrome, and has a good long-term therapeutic effect.

Acupuncture Therapy ; methods ; Adult ; Female ; Humans ; Ovulation ; Polycystic Ovary Syndrome ; physiopathology ; therapy ; Pregnancy ; Pregnancy Rate

OBJECTIVETo observe the effect of Compound Zhajin Granule (CZG) on Toll-like re-ceptor 4 (TLR4) signaling pathway in high-fructose corn syrup induced NASH mice.

METHODSThirty 6-week-old male C3H mice were divided into the high fat and high fructose (HFHFr) group (n = 20) and the control group (n = 10) according to body weight. Mice in the HFHFr group ate high fat diet and drank 20% fructose water, while those in the control group ate common diet and drank common water. After 8 weeks mice in the HFHFr group were divided into two group according to body weight, the HFHFr group and the CZG group, 10 in each group. Mice in the CZG group were fed with high fat forage and 20% fructose water, and administered with 50 mL/kg 12. 8% CZG (prepared by hawthorn, Radix Curcumae, Alisma Orientale, Fritillaria Thunbergii, Silybum Marianum, peach seed in the ratio of 3:1.5:1.5:2:1.5:2:1) by gastrogavage. Mice in the HFHFr group were fed in the same way and daily administered with equal volume of distilled water by gastrogavage. Sixteen weeks later all mice were sacrificed. Body weight, liver wet weight, liver function, and lipid metabolism were detected. Pathological changes of liver tissues were assessed by HE staining, oil red O staining, and Masson staining. Expressions of TLR4, myeloid differentiation factor 88 (MyD88), tumor necrosis factor-alpha (TNF-α) were detected using immunohistochemical staining and real-time fluorescent quantitative PCR.

RESULTSBody weight, alanine aminotransferase (ALT), aspartate aminotransferase (AST) were obviously lower in the CZG group than in the HFHFr group (P < 0.05); oil red O stained area and density were decreased more in the CZG group than in the control group. HE staining showed ballooning inflammation was reduced more in the CZG group than in the HFHFr group. Masson staining was negative. Positive rates of TLR4 and MyD88 and mRNA expressions were significantly lower in the CZG group than in the HFHFr group (all P < 0.05).

CONCLUSIONCZG could significantly inhibit TLR4 signaling pathway of liver in NASH mice.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Diet, High-Fat ; Drugs, Chinese Herbal ; pharmacology ; Fructose ; administration & dosage ; adverse effects ; Inflammation ; Lipid Metabolism ; Male ; Mice ; Mice, Inbred C3H ; Myeloid Differentiation Factor 88 ; metabolism ; Non-alcoholic Fatty Liver Disease ; drug therapy ; Signal Transduction ; drug effects ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo study whether hematologic malignancy patients with anemia have a lower erythropoietin (EPO) response.

METHODSSerum EPO levels were detected by ELISA in patients with hematologic malignancies and with iron deficiency anemia (IDA). Eighty patients with hematologic malignancies, including 13 multiple myeloma (MM), 7 chronic lymphocytic leukemia (CLL) and 60 non-Hodgkin's lymphoma (NHL) were studied. Thirty of them had anemia(21 NHL,6 MM and 3 CLL). Twenty patients with IDA were studied as the control.

RESULTSHematologic malignancy patients with anemia had higher EPO levels [(97.8 +/- 183.9) IU/L] than those with normal Hb values [(27.8 +/- 85.4) IU/L; P <0.01]. In patients with IDA, serum EPO response was inversely correlated with Hb level (r= -0.5, P <0.05) , but no such inverse correlation was found in the hematologic malignancy patients with anemia (r = -0.14). After corrected for Hb level, the serum EPO levels were significantly lower in anemic patients with hematologic malignancies than in IDA patients (P = 0.032) , indicating a decreased EPO response in the former group.

CONCLUSIONAnemia associated with hematologic malignancy might result from an inappropriately low EPO response. EPO treatment for these patients may be beneficial.

Adolescent ; Adult ; Aged ; Anemia, Iron-Deficiency ; blood ; complications ; Enzyme-Linked Immunosorbent Assay ; Erythropoietin ; blood ; Female ; Hematologic Neoplasms ; blood ; complications ; Hemoglobins ; metabolism ; Humans ; Male ; Middle Aged ; Prospective Studies

The human basic Krüppel-like factor (hBKLF) is a newly cloned human transcription factor from the cDNA library of fetal liver. It belongs to the Krüppel-like transcription factor family. Previous expression study showed that it is a hematopoietic related factor. This study was aimed to investigate the effect of hBKLF on cell proliferation, differentiation and hemoglobin synthesis by using K562 cell line as model. The sense and antisense expression plasmids of hBKLF were constructed, and transfected into K562 cells by lipofectamine. After G418 selection for 4 weeks, the cell line with stable expression of the gene was obtained. Then the hBKLF expression level, proliferation ability, colony formation and hemoglobin production were detected by RT-PCR and Western blot, MTT method, methyl cellulose semisolid culture method and benzidine test respectively. The morphologic change of cell was observed with inverted microscope. The results showed that the sense plasmid could increase hBKLF level and antisense plasmid could decrease hBKLF expression. When hBKLF level was down-regulated, K562 cells could proliferate more quickly and synthesize more hemoglobin. But there were no differences in colony formation ability and no apparent morphologic change. It is concluded that hBKLF can inhibit hematopoietic cell proliferation and hemoglobin synthesis. It is suggested that hBKLF plays an important role in the proliferation and differentiation of hematopoietic cells.
Animals ; COS Cells ; Cell Differentiation ; physiology ; Cell Proliferation ; drug effects ; Cell Transformation, Neoplastic ; drug effects ; Cercopithecus aethiops ; Hemoglobins ; biosynthesis ; Humans ; K562 Cells ; Kruppel-Like Transcription Factors ; biosynthesis ; genetics ; pharmacology ; Transcription Factors ; biosynthesis ; genetics ; Transfection

To explore the possibility of a new therapeutic strategy for leukemia by intervening in the DNA methylation to re-express p15 suppressor gene, methylation inhibitors, 5-Aza-2'-deoxycytidine (5-Aza-CdR) and cell differentiation agent (CDAII) were used to treat myelogenous leukemia cell line KG1a in which p15 gene expression was suppressed due to DNA hypermethylation. The biological characteristics of KG1a cells untreated or treated with the agents were investigated and analyzed using morphology, methylation specific-PCR (MSP), (3)H-labeled microassay technique, restriction endonuclease reaction, flow cytometry and immunofluorescence methods. The results indicated that both agents showed concentration-dependent and time-dependent inhibition of cell proliferation. 5-Aza-CdR and CDAII induced apoptosis and cell differentiation with G(2) and G(0)/G(1) arrest respectively. Furthermore, DNA methyltransferase activity and level of methylation in genomic DNA were decreased and p15 protein was re-expressed partially. It is concluded that it is possible to treat leukemia by intervening in the DNA methylation using methyltransferase inhibitors and it is worth to make a thorough study on mechanism of the new strategy.
Azacitidine ; analogs & derivatives ; pharmacology ; Cell Cycle Proteins ; genetics ; Cell Differentiation ; drug effects ; Cell Division ; Cell Line ; Cyclin-Dependent Kinase Inhibitor p15 ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; DNA Methylation ; DNA Modification Methylases ; antagonists & inhibitors ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; pharmacology ; Genes, Tumor Suppressor ; Humans ; Leukemia, Myeloid ; drug therapy ; pathology ; Tumor Suppressor Proteins

This preject is to explore the reversal efficacy of calmodulin antagonist berbamine (BBM) on multidrug resistance (MDR) and its mechanism. Human erythroleukemic cell line K562 and its adriamycin-resistant counterpart K562/A02 were used in the study. The cells were co-cultured with ADR and BBM in different concentrations. MTT assay was used to analyze the effect of BBM on cell growth inhibition. According to the MTT assay, the 50% inhibitory concentration (IC(50)), the multiples of drug resistance and increased sensitivity of ADR were calculated. The concentration of intracellular ADR and expression level of P-gp were detected by flow cytometry (FCM). The expression level of mdr1 mRNA and survivin mRNA was detected by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) with beta-actin as internal reference. The results showed that IC(50) of ADR in K562 and K562/A02 cells was 1.16 +/- 0.09 micro mol/L and 37.47 +/- 1.76 micro mol/L, respectively. The resistant multiple of K562/A02 cells to ADR was 32.30 higher than that of K562 cells. BBM increased the chemo-sensitivity of ADR in K562/A02 cells with dose-dependent relationship, i.e. when 5, 10 and 20 micro mol/L BBM was added in the culture the chemo-sensitivity of ADR was increased to 2.01-, 9.68-, and 41.18-fold (P < 0.01), respectively. After treating K562/A02 cells by 5 or 10 micro mol/L BBM for 2 hours the accumulation of intracellular ADR was increased to 1.41- and 1.52-fold (P < 0.01), respectively. Treating by BBM for 72 hours decreased 4.12% (P < 0.05) and 27.09% (P < 0.01) of P-gp expression, respectively, meanwhile down-regulated expression of mdr1 mRNA and survivin mRNA was found. In conclusion, BBM could increase intracellular concentration of ADR in K562/A02 that down-regulated expression level of mdr1 mRNA and P-gp and survivin so that the sensitivity of K562/A02 to ADR was increased significantly.
ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Alkaloids ; Benzylisoquinolines ; pharmacology ; Calmodulin ; antagonists & inhibitors ; Cell Division ; drug effects ; Doxorubicin ; pharmacokinetics ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Genes, MDR ; Humans ; Inhibitor of Apoptosis Proteins ; K562 Cells ; Leukemia ; drug therapy ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; RNA, Messenger ; analysis

OBJECTIVETo plot a hour-specific transcutaneous bilirubin (TCB) nomogram for healthy neonates, and to evaluate its value for prediction of the risk of neonatal hyperbilirubinemia.

METHODSA total of 5,250 healthy full-term or near-term neonates (gestational age≥35 weeks, birth weight≥2 000 g) were enrolled as subjects. Their TCB values were continuously recorded for 168 hours after birth. The TCB values in the high-risk zones of three time periods, 24-48, 49-72, and 73-96 hours after birth, were used as predictors. The hour-specific TCB nomogram combined with the receiver operating characteristic (ROC) curve was used to evaluate the predictive value of hour-specific TCB nomogram for hyperbilirubinemia.

RESULTSAccording to the hour-specific TCB nomogram, the TCB value dramatically increased during 16-72 hours after birth, and the increase slowed down gradually during 72-144 hours. Finally, the curve reached a plateau after 144 hours. Particularly, the P95 of TCB had been stabilized at 96 hours. The P40, P75, and P95 peak values of TCB were 173, 217, and 248 µmol/L, respectively. For the prediction of hyperbilirubinemia, the areas under the ROC curve of TCB at 24-48, 49-72, and 73-96 hours after birth were 0.77, 0.85, and 0.87, respectively. The high-risk zones at 24-48, 49-72, and 73-96 hours after birth predicted the incidence rates of neonatal hyperbilirubinemia as 35.03%, 43.35%, and 79.95%, respectively, with positive likelihood ratios of 3.35, 4.75, and 22.70, respectively.

CONCLUSIONSThe hour-specific TCB nomogram and the division of TCB risk zones can give a satisfactory prediction of the incidence of neonatal hyperbilirubinemia. The neonate with a bilirubin level in the high-risk zone within 73-96 hours after birth is likely to have hyperbilirubinemia after 73-96 hours.

Bilirubin ; analysis ; Female ; Humans ; Hyperbilirubinemia, Neonatal ; diagnosis ; Infant, Newborn ; Male ; Neonatal Screening ; methods ; Nomograms ; ROC Curve