1.Experimental study of puerarin injection on the hemorheology in acute blood-stasis model rats.
Hong-ping PAN ; Jia-zhen YANG ; Lu-li LI ; Fei YI ; Zhuo-qun HUANG ; Kai-wen HUANG
China Journal of Chinese Materia Medica 2003;28(12):1178-1180
OBJECTIVETo observe the effect of Puerarin Injection on the hemorheology in acute blood-stasis model rats.
METHODThe acute blood-stasis model rats were made by being soaked in ice water afer being injected adrenaline hydrochloride injection in a major dose. The changes of viscosity of whole blood and plasma, blood yield stress, erythrocyte aggregation and the maximum rate of platelet aggregation in the acute blood-stasis model rats were measured with Auto-Viscometer, and then the influence of Puerarin Injection on the hemorheology in the model rats was investigated.
RESULTThe viscosity of whole blood and plasma, and blood yield stress in the acute blood-stasis model rats were significantly higher than those in the normal control group (P < 0.01, P < 0.05). Both the high dose and the low dose of Puerarin Injection could reduce the viscosity of whole blood and plasma, blood yield stress and the maximum rate of platelet aggregation in the acute blood-stasis model rats (P < 0.01, P < 0.05). The high dose could also reduce the erythrocyte aggregation and the deformed Index of red blood cell (P < 0.05).
CONCLUSIONPuerarin Injection can ameliorate the hemorheology in acute blood-stasis model rats, and it has a dose-response relationship.
Animals ; Blood Coagulation Disorders ; blood ; Blood Viscosity ; drug effects ; Dose-Response Relationship, Drug ; Erythrocyte Aggregation ; drug effects ; Erythrocyte Deformability ; drug effects ; Female ; Hemorheology ; drug effects ; Injections ; Isoflavones ; administration & dosage ; isolation & purification ; pharmacology ; Male ; Plants, Medicinal ; chemistry ; Platelet Aggregation ; drug effects ; Pueraria ; chemistry ; Rats ; Rats, Wistar
2.Antigenic analysis of two chimeric hepatitis B core particles presenting the preS1 neutralizing epitopes.
Qin-Dong SU ; Min-Zhuo GUO ; Yao YI ; Si-Yong CHEN ; Zhi-Yuan JIA ; Xue-Xin LU ; Feng QIU ; Sheng-Li BI
Chinese Journal of Experimental and Clinical Virology 2013;27(5):336-339
OBJECTIVETo construct full-length hepatitis B core particles presenting preS1 aa 21-47 epitope and truncated core particles presenting preS1 aa 37-45 epitope on their surface and compare their antigenicity.
METHODSPreS1 aa21-47 epitope and aa 37-45 epitope were inserted respectively into full-length hepatitis B core (aa 1-183) and truncated HBcAg (aa 1-144), between the 78th (Asp) and 79th (Pro). The genes synthesized after the codon optimization were ligated to the pET43. 1a vector with the same cohesive terminal (NdeI and XhoI) and expressed in the E. coli expression system. The morphology of the proteins of interest were observed by electron microscope and characterized by ELISA and Western Blotting.
RESULTSThe morphology of the virus-like particles were confirmed by electron microscope. H2 were solid particles with a diameter of (31.61 +/- 1.27) nm, while H3 were hollow particles with a diameter of (28.46 +/- 1.16) nm. Statistical analysis showed that H2 is larger than H3 in the diameter (P < 0.01). The antigenicity of the inserted epitopes and carrier protein were identified by ELISA and Western Blotting.
CONCLUSIONChimeric hepatitis B core particles presenting the preS1 neutralizing epitopes on their surface have been expressed, purified and identified, which lays the foundation for its application in vaccine research.
Epitopes ; chemistry ; genetics ; immunology ; Hepatitis B ; immunology ; virology ; Hepatitis B Core Antigens ; chemistry ; genetics ; immunology ; Hepatitis B Surface Antigens ; chemistry ; genetics ; immunology ; Hepatitis B virus ; chemistry ; genetics ; immunology ; Humans ; Neutralization Tests ; Protein Precursors ; chemistry ; genetics ; immunology ; Recombinant Fusion Proteins ; chemistry ; genetics ; immunology
3.Prospective clinical study of radix astragali and its compound prescription for treatment of β-thalassemia in children.
Zhuo-Ming LU ; Xin-Hua QIAN ; Zhi-Wen CHEN ; Chun-Hong ZHANG ; Li-Shan GUO ; Jia CHEN
Chinese Journal of Contemporary Pediatrics 2012;14(5):344-349
OBJECTIVETo evaluate the efficacy and safety of radix astragali and its compound prescription for treatment of β-thalassemia in children.
METHODSThis study was a randomized, controlled, double-blind clinical trial. Fifty-seven children with β-thalassemia were randomly assigned to radix astragali, compound prescription (radix astragali+ codonopsis pilosula + tortoise plastron) and placebo control groups after stratifying the patients according to disease type (intermedia and major). The parameters of hematology and safety were assessed after 12 weeks of treatment.
RESULTSAfter 12 weeks of treatment, the mean Hb elevation levels in children with β-thalassemia intermedia from the compound prescription and the radix astragali groups were 1.21±1.12 and 1.05±0.80 g/dL respectively compared with -(0.28±0.51) g/dL in the placebo control group (P<0.01). Mean Hb levels in the compound prescription and radix astragali groups were significantly higher than in the placebo control group (P<0.05). Therapy with both radix astragali and its compound prescription increased fetal hemoglobin, red blood cell, mean corpuscular hemoglobin and reticulocyte levels in children with β-thalassemia intermedia. The total effective rates were 64% and 62% in children with β-thalassemia intermedia from the compound prescription and radix astragali groups respectively, which was significantly higher than in the placebo control group (9%; P<0.01). Therapy with radix astragali or its compound prescription in children with β-thalassemia major had similar but less favourable effects than the same therapy in children with β-thalassemia intermedia. White blood cell, neutrophil, platelet and hepatic and renal functions were not adversely affected by the medicines.
CONCLUSIONSTherapy with radix astragali or its compound prescription is effective and safe in children with β-thalassemia.
Astragalus Plant ; adverse effects ; Child ; Child, Preschool ; Double-Blind Method ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Hemoglobins ; analysis ; Humans ; Male ; Prospective Studies ; beta-Thalassemia ; blood ; drug therapy
4.The extracellular domain of human delta-like-1 expressed and purified from CHO cells promotes expansion of hematopoietic progenitor cells.
Zhuo-Zhuang LU ; Chu-Tse WU ; Hong-Jun LIU ; Qun-Wei ZHANG ; Xiang-Xu JIA ; Li-Sheng WANG
Journal of Experimental Hematology 2003;11(3):222-226
Notch signal path plays important roles in the regulation of proliferation and differentiation of hematopoietic stem cells. An extracellular domain of human Delta-like-1 (hDll-1(ext)), one of Notch ligands, was cloned and expressed in CHO cells, and the effect of hDll-1(ext) on expansion of hematopoietic stem/progenitor cells was investigated in this study. Total RNA was isolated from human marrow mononuclear cells. hDll-1(ext) was amplified by RT-PCR and cloned to T vector, then the gene was sequenced and subcloned to pcDNA3.1/Myc-His(+)A expression vector. The constructed plasmid was transfected into CHO cells with lipofectin and the expression of secreted hDll-1(ext) in G418-resistant clones was assayed by Western blot. hDll-1(ext) high-expressed clone was cultured to collect supernatant. Fusion protein hDll-1(ext) was purified from the supernatant by immobilized metal affinity chromatography (IMAC). The results showed that expression of Notch-1 receptor was detected in cord blood-derived CD34(+) cells by RT-PCR. Human umbilical blood CD34(+) cells were cultured in serum-free medium containing SCF, IL-3, VEGF, and with or without purified hDll-1(ext) for 4 or 8 days. Effect of hDll-1(ext) on the expansion of progenitor cells was analyzed then by clonogenic assays. The number of CFU-Mix and HPP-CFC generated from the culture system containing hDll-1(ext) was 1.5 times of that from the control. In conclusion, the recombinant hDll-1(ext) promotes the expansion of primitive hematopoietic progenitors.
Animals
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Antigens, CD34
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immunology
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Binding Sites
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genetics
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CHO Cells
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Cell Division
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drug effects
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physiology
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Colony-Forming Units Assay
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Cricetinae
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Endothelial Growth Factors
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pharmacology
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Fetal Blood
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cytology
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immunology
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metabolism
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Gene Expression
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Genetic Vectors
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genetics
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Glycoproteins
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genetics
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pharmacology
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physiology
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Hematopoietic Stem Cells
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cytology
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drug effects
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Humans
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Intercellular Signaling Peptides and Proteins
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pharmacology
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Interleukin-3
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pharmacology
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Lymphokines
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pharmacology
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Membrane Proteins
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genetics
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RNA
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genetics
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metabolism
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Receptor, Notch1
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Receptors, Cell Surface
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Recombinant Proteins
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isolation & purification
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pharmacology
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Reverse Transcriptase Polymerase Chain Reaction
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Stem Cell Factor
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pharmacology
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Transcription Factors
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Transfection
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Vascular Endothelial Growth Factor A
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Vascular Endothelial Growth Factors
5.Preclinical evaluation of pseudoallergic reactions on Chinese herbal injections: study on animal strain and gender difference.
Yu-shi ZHANG ; Yan YI ; Chun-ying LI ; Yong ZHAO ; Lian-mei WANG ; Jia-yin HAN ; Yu-ting LU ; Jing-zhuo TIAN ; Xiao-yi FENG ; Gui-qin LI ; Ai-hua LIANG
China Journal of Chinese Materia Medica 2015;40(14):2717-2722
Pseudoallergic reactions occured after the first administration of patients, and the pathogenic mechanisms of them were different from the allergic reactions which needed excitation after antigen sensitization. To provide a basis for evaluation, clinical use and drug development of pseudoallergic reactions, the models were established by two kinds of Chinese herbal injections (CHI) both on different strain or gender mice. With the use of ICR, Kunming, BALB/C, C57 mice, pseudoallergic tests of two CHI were conducted to compare the sensitivity of four strains mice, and compared the differences in male and female animals. Test substances contain 0.8% Evans blue (EB) were intravenously injected into different strain and gender mice. Scores of ear blue staining and quantitation of ear EB exudation were the parameters for pseudoallergic reaction. Results of strain difference indicated that both CHI A and B could cause severe pseudoallergic reactions indicated by obvious vascular hyperpermeability on ICR mice. The pseudoallergic reactions in ICR mice are more obvious under the the same dose of injection, which stated the sensibility of ICR mice. And the reactions of KM mice and BALB/C mice were slightly reduced which compared to ICR mice, even alomost nothing on C57 mice. Comparison results of gender difference showed that one CHI was not have significant difference in male and female animals, but male animals were more susceptible than females on another CHI. Therefore, ICR mice were preferable experimental strain on the model of pseudoallergic reactions induced by CHI A and B. Because of female animals were easily influenced by estrous cycle, the pseudoallergic reactions induced by CHI A and B select and use male mice befittingly.
Animals
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Drug Hypersensitivity
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etiology
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Drugs, Chinese Herbal
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adverse effects
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Female
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Injections
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Male
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Mice
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Mice, Inbred BALB C
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Mice, Inbred ICR
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Sex Characteristics
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Species Specificity
6.Effect of type I collagen on the adhesion, proliferation, and osteoblastic gene expression of bone marrow-derived mesenchymal stem cells.
Gang LIU ; Yun-Yu HU ; Jian-Ning ZHAO ; Su-Jia WU ; Zhuo XIONG ; Rong LU
Chinese Journal of Traumatology 2004;7(6):358-362
OBJECTIVETo investigate the effects of porous poly lactide-co-glycolide (PLGA) modified by type I collagen on the adhesion, proliferation, and differentiation of rabbit marrow-derived mesenchymal stem cells (MSCs).
METHODSThe third generation MSCs isolated from mature rabbits by density gradient centrifugation were cultured at different initial concentrations on 0.3 cm x 1.2 cm x 2.0 cm 3-D porous PLGA coated by type I collagen in RPMI 1640 containing 10% fetal calf serum, while cultured on PLGA without type I collagen as control. The cells adhesive and proliferative behavior at 7, 14, and 21 days after inoculation was assessed by determining the incorporation rate of [(3)H]-TdR. In order to examine MSCs differentiation, the expression of osteoblasts marker genes, osteocalcin (OCN), alkaline phosphatase (ALP), osteopontin (OPN) mRNA, were evaluated by reverse transcription-polymerase chain reaction (RT-PCR), and further more, the cell morphology at 21 days was also observed by scanning electron microscope (SEM).
RESULTSType I collagen promoted cell adhesion on PLGA. The valve was significantly higher than controls (6 h, 2144 cpm+/-141 cpm vs. 1797 cpm+/-118 cpm, P=0.017; 8 h, 2311 cpm+/-113 cpm vs. 1891 cpm+/-103 cpm, P=0.01). The cells which cultured on PLGA coated with type I collagen showed significantly higher cell proliferation than controls on the 7 th day (1021 cpm+/-159 cpm vs. 451 cpm+/-67 cpm, P=0.002), the 14th day (1472 cpm+/-82 cpm vs. 583 cpm+/-67 cpm, P<0.001) and 21 th day (1728 cpm+/-78 cpm vs. 632 cpm+/-55 cpm, P<0.001). Osteoblasts markers, OCN, ALP, OPN mRNA, were all detected on PLGA coated by type I collagen on the 21 th day, but OCN, OPN mRNA could not be found in controls. Spindle and polygonal cells well distributed on the polymer coated by type I collagen while cylindric or round cells in controls.
CONCLUSIONSType I collagen is effective in promoting the adhesion, proliferation and differentiation of MSCs on PLGA.
Biocompatible Materials ; pharmacology ; Cell Adhesion ; Cell Proliferation ; Collagen Type I ; pharmacology ; Gene Expression ; Humans ; Lactic Acid ; pharmacology ; Mesenchymal Stromal Cells ; physiology ; Osteoblasts ; physiology ; Polyglycolic Acid ; pharmacology ; Polymers ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Engineering
7.Hepatocyte growth factor recruits endothelial progenitor cells from bone marrow into blood circulation.
Qun-wei ZHANG ; Hong-jun LIU ; Hai-feng DUAN ; Xiao-qin HA ; Hua WANG ; Xiang-xu JIA ; Zhuo-zhuang LU ; Chu-tse WU ; Li-sheng WANG
Chinese Journal of Applied Physiology 2005;21(1):100-103
AIMTo assess whether hepatocyte growth factor recruits bone marrow-derived endothelial progenitor cells into blood circulation to participate in postnatal angiogenesis and endothelium repair.
METHODSThe adenovirus vector encoding HGF gene (Ad-HGF) were intravenous administrated into BALB/c mice, and then serum HGF was determined by enzyme-linked immunosorbent assay, the number of CD34+ cells in peripheral blood was assayed by flow cytometry, and the nucleated cells in peripheral blood were isolated, cultured and the endothelial cell colonies were characterized by staining with antibodies against tie-2, vWF. The carbon tetrachloride-induced liver damage model of female mice was established. The peripheral blood nucleated cells of Ad-HGF treated male mice were intravenous administrated into these mice, and 4 weeks later, in situ hybridization for the sry gene was used to identify the implanted cells in the damaged tissues.
RESULTSIntravenous administration of Ad-HGF resulted in significant elevation of serum hepatocyte growth factor level and induced profoundly increase of endothelial progenitor cells in the peripheral blood, which were characterized by their ability to form endothelial cell colonies in culture and expression of CD34, tie-2, and vW factor. HGF-mobilized endothelial progenitors could incorporate into sites of neovascularization in a liver regeneration model.
CONCLUSIONHepatocyte growth factor could markedly recruit bone marrow-derived endothelial progenitor cells into blood circulation.
Animals ; Bone Marrow Cells ; cytology ; Endothelial Cells ; cytology ; Female ; Hematopoietic Stem Cell Mobilization ; Hepatocyte Growth Factor ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Stem Cells ; cytology
8.Suppressive effect of Notch signal activation on apoptosis of multiple myeloma cells.
Xiang-Xu JIA ; Zhuo-Zhuang LU ; Hua WANG ; Hai-Feng DUAN ; Qun-Wei ZHANG ; Chu-Tse WU ; Li-Sheng WANG
Journal of Experimental Hematology 2004;12(3):335-339
The proliferation and apoptosis of multiple myeloma (MM) cells were regulated by bone marrow microenvironments in which Notch signal plays important role in mediating cell-cell communication. However, the regulatory effect of Notch signal on the proliferation and apoptosis of multiple myeloma cells remains unclear. In this study, regulatory effect of Notch signal on the apoptosis of MM cells induced by DMS (N, N-dimethylsphingosine) was investigated. RT-PCR was used to identify the expression of Notch receptor and related molecules such as Dll-1, Jagged-1, Deltex-1 in MM cell lines. The intracellular domain of Notch (ICN), active form of Notch, was transferred into MM cells by retrovirus. The apoptosis of MM cells was determined by trypan blue exclusion tests and TdT-mediated dUTP nick end labeling (TUNEL) assay. The results showed that multiple myeloma cells expressed the Notch-1 and its related molecules. Notch activated multiple myeloma cell lines were obtained. Activation of Notch protected the multiple myeloma cells from the apoptosis induced by DMS,which was determined by cell viability and TUNEL assay. In conclusion, Notch signal suppressed the apoptosis of multiple myeloma cells and would possibly be a novel therapeutic target.
Apoptosis
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Cell Division
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Humans
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Multiple Myeloma
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drug therapy
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pathology
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Receptor, Notch1
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Receptors, Cell Surface
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physiology
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Signal Transduction
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Transcription Factors
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physiology
9.Expression of heparanase and its coagulation proteins on the surface of leukemic cells
Dong-xia ZHANG ; Zhi-qin LI ; Yan YUN ; Xuan-mao HAN ; Qi-tu HE ; Guo-rong JIA ; Yan LU ; Hong-jie MA ; Xue-wen LIU ; Xue-qin BAI ; Mei-xiang GUO ; Li-xia ZHUO
Journal of Leukemia & Lymphoma 2011;20(12):723-725,729
Objective To explore whether the expression level of heparanase (HPA) and its coagulation proteins on leukemic blast membrane could determine the hemostatic balance on the surface of leukemia cells.Methods Forty patients of leukemia were studied,and 20 patients with iron dificient anemia as the control group.Expression of tissue factor (TF),heparanase (HPA),tissue factor pathway inhibitor (TFPI),and urokinase plasminogen activator receptor (UPAR) on leukemic blast surfaces were analyzed by flowcytometry.Results The expression of TF,UPAR,and HPA in AML,ALL,CML,CLL and CRAL groups were significantly higher compared with the control group (t =.3.289,3.507,2.701,P <0.05; t =2.498,0.802,3.090,P <0.05; t =2.642,3.308,2.696,P <0.05; t =3.417,3.434,2.382,P <0.05; t =2.193,2.272,2.263,P <0.05).There were no significantly differences between the leukemic cell expression of TFPI and the control group (P >0.05).Expression of TF,UPAR,HPA in AML patients were significantly higher than ALL,CML and CLL groups (t =2.463,2.179,2.276,P <0.05; t =2.637,2.402,2.095,P <0.05; t =2.548,2.425,2.412,P <0.05).The levels of TF,UPAR and HPA in M3,M4 and M5 patients were higher than that of M1,M2 groups (P <0.05).There were no significantly differences among M3,M4 and M5 (P >0.05).Conclusions These results suggest that TF,UPAR and HPA are predominately expressed on leukemic blast surface,particularly in M3and M4,5 subtypes.The expression of coagulation proteins on blast membrane might determine the hemostatic balance on the surface of leukemia cells.
10.Regulatory Effect of Huadu Sanyinfang on PI3K/Akt/NF-κB Pathway in Triple-negative Breast Cancer Patients with Qi-deficiency Constitution: An Exploration Based on Differential Expression of miRNA
Yong-jia CUI ; Zhi-li ZHUO ; Wen-ping LU
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(22):194-200
Objective:To explore the regulatory effect of Huadu Sanyinfang on phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt)/transcription factor nuclear factor-