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1.
Small ; 15(24): e1901848, 2019 Jun.
Article in English | MEDLINE | ID: mdl-31062504

ABSTRACT

The zinc-bromine flow battery (ZBFB) is one of the most promising technologies for large-scale energy storage. Here, nitrogen-doped carbon is synthesized and investigated as the positive electrode material in ZBFBs. The synthesis includes the carbonization of the glucose precursor and nitrogen doping by etching in ammonia gas. Physicochemical characterizations reveal that the resultant carbon exhibits high electronic conductivity, large specific surface area, and abundant heteroatom-containing functional groups, which benefit the formation and exposure of the active sites toward the Br2 /Br- redox couple. As a result, the assembled ZBFB achieves a voltage efficiency of 83.0% and an energy efficiency of 82.5% at a current density of 80 mA cm-2 , which are among the top values in literature. Finally, the ZBFB does not yield any detectable degradation in performance after a 200-cycle charging/discharging test, revealing its high stability. In summary, this work provides a highly efficient electrode material for the zinc-bromine flow battery.

2.
Sleep Med Rev ; 29: 108-18, 2016 10.
Article in English | MEDLINE | ID: mdl-26866454

ABSTRACT

The main pharmacological effects of sedative agents are sedation, hypnosis, antianxiety, and antidepression. Traditional Chinese medicine (TCM) has a long history of clinical experience in treating insomnia. This review focuses mainly on the role of active ingredients from TCM in the treatment of insomnia. Single herbs and their active ingredients from TCM with hypnotic effects are summarized through reviewing the relevant literature published in the past 20 y. The active ingredients are divided into alkaloids, terpenoids, and volatile oils, flavonoids, lignanoids and coumarins, saponins, and others. Current studies on TCM in treating insomnia are described from the aspects of active ingredients, sources, experimental models and methods, results, and mechanisms. In addition, Chinese compound prescriptions developed from a variety of single herbs with sedative-hypnotic effects are introduced. The acting pathways of TCM are covered from the perspectives of regulating central neurotransmitters, influencing sleep-related cytokines, and improving the structure of the central nervous system.


Subject(s)
Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacology , Hypnotics and Sedatives/chemistry , Hypnotics and Sedatives/pharmacology , Medicine, Chinese Traditional , Central Nervous System/drug effects , Central Nervous System/physiology , Humans , Sleep/drug effects , Sleep/physiology
3.
Chin J Nat Med ; 13(4): 299-302, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25908628

ABSTRACT

The present study was designed to isolate and characterize novel chemical constituents of the stem bark of Juglans mandshurica Maxim. (Juglandaceae). The chemical constituents were isolated and purified by various chromatographic techniques. The structures of the compounds were elucidated on the basis of spectral data (1D and 2D NMR, HR-ESI-MS, CD, UV, and IR) and by the comparisons of spectroscopic data with the reported values in the literatures. Two long chain polyunsaturated fatty acids (1 and 2) were obtained and identified as (S)-(8E,10E)-12-hydroxy-7-oxo-8,10-octadecadienoic acid (1) and (S)-(8E, 10E)-12-hydroxy-7-oxo-8,10-octadecadienoic acid methyl ester (2). To the best of our knowledge, this is the first report on the isolation and structural elucidation of the two new conjugated ketonic fatty acids from this genus.


Subject(s)
Fatty Acids, Unsaturated/isolation & purification , Juglans/chemistry , Plant Bark/chemistry , Fatty Acids, Unsaturated/chemistry , Spectrum Analysis
4.
J Neural Transm (Vienna) ; 122(8): 1105-13, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25596842

ABSTRACT

It has long been known that the polymorphisms of angiotensin-converting enzyme gene (ACE) are associated to increase risk of Alzheimer's disease (AD) in Chinese population. However, consistent results were not obtained among studies. This study is aimed to clarify the association between ACE insertion (I)/deletion (D) polymorphism (rs1799752) and AD. Literatures were searched from PubMed, Embase, Cochrane Library, CNKI, Wanfang and VIP databases without language restrictions. Eleven separate studies were suitable for the inclusion criterion. The selected studies contained 2,763 Chinese participants, including 1,383 in AD group and 1,380 controls. Pooled odds ratios (ORs) were calculated to assess the association between ACE I/D polymorphism and AD. Our case-control data indicated that ACE insertion is a risk allele in all genetic models: additive model (I vs. D: OR = 1.32, 95 % CI 1.07-1.62, P = 0.008), dominant model (II + ID vs. DD: OR = 1.61, 95 % CI 1.08-2.41, P = 0.02) and recessive model (II vs. ID + DD: OR = 1.39, 95 % CI 1.07-1.81, P = 0.01). Heterogeneity between studies was significant (P < 0.10) but not in stratification defined by the selection of controls (P > 0.10). After stratification according to the selection of controls, the carrier of ACE I allele remained a high risk for AD in population-based samples subgroup (I vs. D: P = 0.008, OR = 1.32, 95 % CI 1.07-1.61, P(heterogeneity) = 0.47, I (2) = 0 %). Our study provided solid evidence suggesting that ACE gene I/D polymorphism is a genetic risk factor for AD in Chinese population.


Subject(s)
Alzheimer Disease/genetics , Asian People/genetics , Genetic Predisposition to Disease , INDEL Mutation , Polymorphism, Genetic , Alzheimer Disease/ethnology , Case-Control Studies , China , Humans
5.
Food Funct ; 5(7): 1369-73, 2014 Jul 25.
Article in English | MEDLINE | ID: mdl-24818222

ABSTRACT

Curcuma phaeocaulis Val. has been used as a health food in China for a long time. This research aimed to isolate and identify its active compounds with protective effects against hydrogen peroxide-induced PC12 cell death. 70% ethanol extracts of C. phaeocaulis were re-extracted and three fractions of water, petroleum ether and ethyl acetate were obtained. Three diphenylheptane compounds from the ethyl acetate fraction were identified for the first time from C. phaeocaulis, and compound III was considered to be a new structure. All of the three compounds displayed certain protective effects against toxicity in PC12 cells. For all concentrations, compound III displayed a more significant protective effect than ethanol extracts, the ethyl acetate fraction, and the other two compounds. At a concentration of 50 µg mL(-1), the survival rate of damaged PC-12 cells treated with compound III reached 84.7%. Diphenylheptanes were concluded to be the main compounds responsible for the health effects of C. phaeocaulis.


Subject(s)
Curcuma/chemistry , Heptanes/pharmacology , Hydrogen Peroxide/adverse effects , Plant Extracts/pharmacology , Animals , PC12 Cells , Protective Agents/pharmacology , Rats
6.
Oncol Rep ; 32(1): 293-301, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24842472

ABSTRACT

Our previous study found that caveolin-1 (CAV-1) protein expression is upregulated during oleanolic acid (OA)-induced inhibition of proliferation and promotion of apoptosis in HL-60 cells. CAV-1 is the main structural protein component of caveolae, playing important roles in tumorigenesis and tumor development. It has been shown that cav-1 expression is lower in leukemia cancer cell lines SUP-B15, HL-60, THP-1 and K562 and in chronic lymphocytic leukemia primary (CLP) cells when compared with normal white blood cells, with the lowest cav-1 expression level found in HL-60 cells. To study the effects of cav-1 in HL-60 cells and the effects of cav-1 overexpression on OA drug efficacy, cav-1 was overexpressed in HL-60 cells using lentiviral-mediated transfection combined with OA treatment. The results showed that cav-1 overexpression inhibited HL-60 cell proliferation, promoted apoptosis, arrested the cell cycle in the G1 phase and inhibited activation of the PI3K/AKT/mTOR signaling pathway. Overexpression of CAV-1 also increased HL-60 cell sensitivity to OA. To further verify whether OA affects HL-60 cells via the activation of downstream signaling pathways by CAV-1, cav-1 gene expression was silenced using RNAi, and the cells were treated with OA to examine its efficacy. The results showed that after cav-1 silencing, OA had little effect on cell activity, apoptosis, the cell cycle and phosphorylation of HL-60 cells. This study is the first to show that CAV-1 plays a crucial role in the effects of OA on HL-60 cells.


Subject(s)
Caveolin 1/genetics , Caveolin 1/metabolism , Leukemia/pathology , Oleanolic Acid/pharmacology , Apoptosis/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Gene Expression Regulation, Neoplastic/drug effects , Gene Silencing , HL-60 Cells , Humans , Leukemia/metabolism , MAP Kinase Signaling System/drug effects , Phosphorylation/drug effects
7.
Food Funct ; 4(12): 1811-8, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24150197

ABSTRACT

Penthorum chinense Pursh is rich in flavonoids, which have strong antioxidant and anticomplement activities. In order to optimize their extraction conditions, various extraction parameters were chosen to identify their effects on flavonoids extraction. Single factor and Box-Behnken experimental designs consisting of 24 experimental runs and five replicates at zero point were applied to obtain the optimal extraction yield. The optimization conditions for flavonoids extraction were determined as follows: ethanol concentration 60.89%, extraction time 68.15 min, temperature 52.89 °C and liquid/solid ratio 19.70 : 1. The corresponding flavonoids content was 7.19%. The regression equation was found to fit well with the actual situation. Furthermore, the antioxidant activity (the free radical scavenging ability and ferric reducing/antioxidant power) and anticomplement ability of the flavonoids from P. chinense were determined. Results showed that the flavonoids of P. chinense displayed significant antioxidant and anticomplement activities. Its antioxidant activity can compete with ascorbic acid (Vc), whereas its anticomplement activity (IC50 = 111.6 µg ml(-1)) surpassed the effect of heparin (IC50 = 399.7 µg ml(-1)) which was used as the positive control, suggesting that P. chinense flavonoids and their related products could potentially be used as a promising natural agent in the treatment of humoral effector inflammation.


Subject(s)
Antioxidants/pharmacology , Complement Inactivator Proteins/chemistry , Complement Inactivator Proteins/pharmacology , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacology , Flavonoids/pharmacology , Magnoliopsida/chemistry , Animals , Antioxidants/chemistry , Antioxidants/isolation & purification , Complement Inactivator Proteins/isolation & purification , Drugs, Chinese Herbal/isolation & purification , Erythrocytes/drug effects , Erythrocytes/physiology , Flavonoids/chemistry , Flavonoids/isolation & purification , Sheep
8.
Zhongguo Zhong Yao Za Zhi ; 38(7): 1067-70, 2013 Apr.
Article in Chinese | MEDLINE | ID: mdl-23847959

ABSTRACT

OBJECTIVE: To observe the effect of Dingguier umbilical paste on rats with functional dyspepsia and mice with splenic asthenia, and investigate the related mechanism. METHOD: Functional dyspepsia models of rats were made by irregular food intake plus diluted hydrochloric acid. Successional treatments were offered for 14 days. The rats weights, contents of serum NO, AChE and MC were measured. The rats with splenic asthenia were made by rhubarb feed, and observed the affection of gastric emptying. RESULT: Compared with those in the model control group, the weight of rats in all dosages Dingguier umbilical paste groups increased obviously (P < 0.05), pepsin activity of rats in the dosage (1.34 g x kg(-1)) Dingguier umbilical paste groups was significantly higher and the contents of NO and quantities of MC in the dosage (2.67 g x kg(-1)) Dingguier umbilical paste groups decreased clearly (P < 0.05), and the contents of serum AChE in all dosages Dingguier umbilical paste groups rose apparently. The weight of mice with splenic asthenia increased obviously, accelerated gastric emptying, and improved the symptom. CONCLUSION: Dingguier umbilical paste has significant improvement of indigestion. The related mechanism may be to reduce the content of serum NO and the quantity of MC and enhance the content of serum AChE.


Subject(s)
Asthenia/drug therapy , Drugs, Chinese Herbal/administration & dosage , Dyspepsia/drug therapy , Spleen/drug effects , Animals , Asthenia/pathology , Asthenia/physiopathology , Body Weight/drug effects , Dyspepsia/physiopathology , Eating/drug effects , Female , Gastric Emptying/drug effects , Humans , Male , Mice , Rats , Rats, Wistar , Spleen/pathology , Umbilicus
9.
Expert Opin Drug Saf ; 11(6): 985-1002, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22992190

ABSTRACT

INTRODUCTION: Traditional Chinese medicine (TCM) is widely used around the world. However, with its wide use has been the identification of a number of toxicological issues that have severely restricted its use in clinical treatment. The identification of these toxic substances within TCM has become somewhat of a hot topic in recent years. AREAS COVERED: This article reviews literature published on professional authoritative journals in the last 10 years on the toxic constituents and toxicology of TCM, including chemical structures, absorption and metabolism. The literature search for this article was based, but not limited to, toxic constituents including: alkaloids, glycosides, toxic proteins, polypeptide, amino acids, phenols or organic acids, terpenes and lactones. The authors discuss the toxic substances referring to their toxicity on organs, tissues and systems. EXPERT OPINION: More and more toxic constituents from different TCMs have been identified, in addition to information on how they act in the body at a molecular level. However, the toxicology of TCMs is very complex, and although some progress has been made, a lot work is still needed in order to put an end to toxic incidents.


Subject(s)
Drugs, Chinese Herbal/adverse effects , Medicine, Chinese Traditional/adverse effects , Animals , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacokinetics , Humans , Medicine, Chinese Traditional/methods
10.
J Food Sci ; 77(9): C975-82, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22900526

ABSTRACT

Eclipta prostrasta L. is a traditional Chinese medicine herb, which is rich in saponins and has strong antiviral and antitumor activities. An ultrasonic-assisted extraction (UAE) technique was developed for the fast extraction of saponins from E. prostrasta. The content of total saponins in E. prostrasta was determined using UV/vis spectrophotometric methods. Several influential parameters like ethanol concentration, extraction time, temperature, and liquid/solid ratio were investigated for the optimization of the extraction using single factor and Box-Behnken experimental designs. Extraction conditions were optimized for maximum yield of total saponins in E. prostrasta using response surface methodology (RSM) with 4 independent variables at 3 levels of each variable. Results showed that the optimization conditions for saponins extraction were: ethanol concentration 70%, extraction time 3 h, temperature 70 °C, and liquid/solid ratio 14:1. Corresponding saponins content was 2.096%. The mathematical model developed was found to fit well with the experimental data. Practical Application: Although there are wider applications of Eclipta prostrasta L. as a functional food or traditional medicine due to its various bioactivities, these properties are limited by its crude extracts. Total saponins are the main active ingredient of E. prostrasta. This research has optimized the extraction conditions of total saponins from E. prostrasta, which will provide useful reference information for further studies, and offer related industries with helpful guidance in practice.


Subject(s)
Eclipta/chemistry , Saponins/isolation & purification , Ultrasonics/methods , Antineoplastic Agents/isolation & purification , Antineoplastic Agents/pharmacology , Antiviral Agents/isolation & purification , Antiviral Agents/pharmacology , Drugs, Chinese Herbal/isolation & purification , Drugs, Chinese Herbal/pharmacology , Plant Extracts/chemistry , Plant Extracts/pharmacology , Reproducibility of Results , Saponins/pharmacology , Temperature , Ultrasonics/instrumentation
11.
Pharm Biol ; 50(2): 225-31, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22235889

ABSTRACT

CONTEXT: The rhizome of Wikstroemia indica (L.) C. A. Mey (Thymelaeaceae) is widespread in China which has been widely used in China as folk medicine for the treatment of syphilis, arthritis, whooping cough, and cancer. Due to its multiactivities, its extract has an attractive potential as a promising natural agent in the pharmaceutical industries. OBJECTIVE: Aims of this study were to optimize the extraction process of the flavonoids from W. indica, and evaluate its multiple activities. MATERIALS AND METHODS: An orthogonal test design was employed to optimize the extraction procedure of flavonoids from W. indica. And multichromatography and spectroscopy were used to study the chemical compounds of W. indica, while several bioactivity assays were used to evaluate the antibacterial, anti-inflammatory, and antitumor activities of W. indica. RESULTS: Optimal extraction conditions were determined: ethanol concentration was 60%; extraction time was 60 min; liquid-solid ratio was 16:1 and the power of ultrasonic instrument was 160 W. Four compounds: daphnoretin, chrysophanol, myricitrime and rutin were purified from W. indica, and chrysophanol was identified from this plant for the first time. The extract of W. indica displayed significant antimicrobial and anti-inflammatory activities. Daphnoretin showed a significant inhibition effect on CNE cells and HeLa cells lines at the concentrations ranging from 15.6 to 125 µg/mL, the tendency of antitumor effect was displayed in a concentration-dependent manner. DISCUSSION AND CONCLUSIONS: Extracts of W. indica could potentially be used as a promising natural agent in the pharmaceutical industries.


Subject(s)
Flavonoids/pharmacology , Plant Extracts/pharmacology , Wikstroemia/chemistry , Anti-Bacterial Agents/administration & dosage , Anti-Bacterial Agents/isolation & purification , Anti-Bacterial Agents/pharmacology , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/isolation & purification , Anti-Inflammatory Agents/pharmacology , Antineoplastic Agents, Phytogenic/administration & dosage , Antineoplastic Agents, Phytogenic/isolation & purification , Antineoplastic Agents, Phytogenic/pharmacology , Cell Line, Tumor , Dose-Response Relationship, Drug , Flavonoids/administration & dosage , Flavonoids/isolation & purification , HeLa Cells , Humans , Medicine, Chinese Traditional , Plant Extracts/administration & dosage , Plant Extracts/chemistry
12.
J Food Sci ; 76(9): C1219-25, 2011.
Article in English | MEDLINE | ID: mdl-22416680

ABSTRACT

UNLABELLED: A comparative study of steam distillation extraction (SDE), reflux extraction (RE), and ultrasound-assisted extraction (USE) was conducted for the extraction of essential oils from the bud of Citrus aurantium L. var. amara Engl. Each method was evaluated in terms of qualitative and quantitative composition of the isolated essential oil by gas chromatography/mass spectrometry (GC/MS). The extract yields of essential oil were 0.16%, 2.18%, and 2.34%, respectively. A total of 82 compounds were identified by GC/MS. The main components obtained by SDE were terpinen-4-ol (20.98%), dipentene (11.67%), terpinene (9.24%), those by RE were palmitic acid (20.61%), 2-chloroethyl linoleate (14.54%), tetracosane (12.26%), and α-linolenic acid (11.24%), and those by USE were tetracosane (11.32%), heneicosane (11.06%), and palmitic acid (8.76%). Comparative analysis indicated that SDE was favorable for the extraction of monoterpene hydrocarbons, sesquiterpene hydrocarbons, alcohols, and carbonyl compounds, RE and USE had certain advantages in the extraction of aliphatic saturated hydrocarbons, organic acids, and esters. It is concluded that different extraction methods may lead to different yields of essential oils; the choice of appropriate method is very important to obtain more desired components with higher physiological activities. PRACTICAL APPLICATION: C. aurantium oils from different plant parts have great economic, medicinal, and nutritional values because of their wide-spectrum biological activities. The essential oil from C. aurantium L. var amara is one of the best C. aurantium oils. The data presented in this article will help us understand the relationship between essential oils and its extraction methods and know more about the aromatic components of Citrus aurantium bud. The methods established in this study will provide useful reference information for further studies, and offer essential oil industries with helpful guidance in practice.


Subject(s)
Citrus/chemistry , Food Handling/methods , Gas Chromatography-Mass Spectrometry/methods , Oils, Volatile/chemistry , Alkanes/analysis , Cyclohexenes/analysis , Limonene , Linoleic Acids/analysis , Menthol/analogs & derivatives , Menthol/analysis , Monoterpenes/analysis , Oils, Volatile/isolation & purification , Palmitic Acid/analysis , Sesquiterpenes/analysis , Terpenes/analysis , alpha-Linolenic Acid/analysis
13.
Curr Med Chem ; 17(36): 4521-37, 2010.
Article in English | MEDLINE | ID: mdl-21062253

ABSTRACT

Traditional Chinese medicine (TCM) has undergone a long history of clinical practice, which can arrive at ideal therapeutic effects by regulating the body's overall function. However, the complex nature of TCM determines a difficult study on the mechanism and material base of TCM. The current investigations of TCM indicate that the development of modern biotechnology will offer a strong arm in the process of the study. This review focused on the application of the modern biotechnology, including transgenic, gene knockout, cell membrane chromatography (CMC), molecular biochromatography (MBC), gene chips, proteomics, etc. in the research of pharmacodynamic effects of TCM at levels of whole animal, cell and molecular models over the past decade. The whole animal models established by the transgenic and gene knockout technology can truly reflect the characteristics of the target gene activity. Thereby the created animal model could share the pathology of maximum degree of approximation. Cellular models are especially suitable for the situation that functional proteins, enzymes, or drug targets are difficult to separate, or the characteristics of the drugs are unidentified. The utilization of MBC can not only achieve high-throughput screening, but also directly detect the chemical composition of the active components relative to the receptors. Based on the remarkable progress of genomics and proteomics and the technique of gene chips, the bioactive components of TCM can be screened through observing the changes of genes or proteins before and after the compounds acting on the cells.


Subject(s)
Cell Membrane/chemistry , Drugs, Chinese Herbal/pharmacokinetics , Medicine, Chinese Traditional , Models, Biological , Animals , Chromatography, High Pressure Liquid , Models, Molecular , Oligonucleotide Array Sequence Analysis
14.
Cardiol Young ; 20(4): 426-32, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20420740

ABSTRACT

OBJECTIVE: To investigate the epidemiological characteristics of Kawasaki disease in Jilin province of China and explore its clinical features. METHODS: The medical records of children with Kawasaki disease hospitalised in the First Affiliated Hospital of Jilin University and Yanbian University between January, 2000 and December, 2008 were retrospectively analysed. RESULTS: A total of 735 children with Kawasaki disease were enrolled in this study with 483 boys and 252 girls. The ratio of male to female was 1.92:1. The ages of the children at onset varied from 51 days to 12 years with a mean age of 2.8 years. The children under the age of 5 years accounted for 79.5%, but most children were 2-3 years old. Kawasaki disease occurred all the year and more frequently in both the ending of spring and the beginning of summer. Fever was the most common clinical feature and enlarged cervical lymph nodes were the smallest clinical feature. A cardiovascular lesion was found in 41.4% of these children, in whom coronary artery dilatation was the most common (26.97%). A total of 117 (18.2%) of 643 children (87.5%) receiving intravenous immunoglobulin had a non-response to gamma globulin. Of the 117 children, 66 (56.4%) had cardiovascular lesion. Kawasaki disease recurred in 19 children (2.6%). CONCLUSION: The incidence of Kawasaki disease in Jilin province has shown an increasing tendency. The age at onset is slightly higher than that described in other reports. Kawasaki disease is the most common in both the ending of spring and the beginning of summer, and the second incidence peak occurs in autumn.


Subject(s)
Mucocutaneous Lymph Node Syndrome/epidemiology , Age Distribution , Age of Onset , Child , Child, Preschool , China , Cohort Studies , Female , Humans , Immunoglobulins, Intravenous/therapeutic use , Immunologic Factors/therapeutic use , Incidence , Infant , Male , Mucocutaneous Lymph Node Syndrome/diagnosis , Mucocutaneous Lymph Node Syndrome/therapy , Retrospective Studies , Seasons , Sex Distribution
16.
Cancer Res ; 66(19): 9453-60, 2006 Oct 01.
Article in English | MEDLINE | ID: mdl-17018600

ABSTRACT

Blood coagulation factor VII (fVII) is physiologically synthesized in the liver and released into the blood. Binding of fVII to tissue factor (TF) at sites of vascular injury triggers coagulation and hemostasis. TF/fVIIa complex formation on the surface of cancer cells plays important roles in cancer biology. Although fVII is synthesized by hepatocellular carcinoma, it remained unclear how TF/fVIIa complex formation and promigratory signaling can occur for most other cancers in extravascular locations. Here, we show by reverse transcription-PCR analysis that nonhepatic cancer cell lines constitutively express fVII mRNA and that endogenously synthesized fVIIa triggers coagulation activation on these cells. fVIIa expression in cancer cells is inducible under hypoxic conditions and hypoxia-inducible factor-2 alpha bound the promoter region of the FVII gene in chromatin immunoprecipitation analyses. Constitutive fVII expression in an ovarian cancer cell line enhanced both migration and invasion. Enhanced motility was blocked by anti-TF antibodies, factor Xa inhibition, and anti-protease-activated receptor-1 antibody treatment, confirming that TF/fVIIa stimulated migration by triggering cell signaling. This study shows that ectopic synthesis of fVII by cancer cells is sufficient to support proinvasive factor Xa-mediated protease-activated receptor-1 signaling and that this pathway is inducible under hypoxia.


Subject(s)
Factor VII/biosynthesis , Neoplasm Invasiveness/physiopathology , Neoplasm Proteins/biosynthesis , Basic Helix-Loop-Helix Transcription Factors/physiology , Blood Coagulation , Carbon-Carbon Ligases/biosynthesis , Carbon-Carbon Ligases/genetics , Cell Hypoxia/genetics , Cell Line, Tumor/metabolism , Cell Movement/physiology , Factor VII/genetics , Factor VII/physiology , Factor Xa/physiology , Female , Humans , Neoplasm Proteins/genetics , Neoplasm Proteins/physiology , Neoplasms/blood , Neoplasms/pathology , Ovarian Neoplasms/pathology , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , RNA, Neoplasm/biosynthesis , RNA, Neoplasm/genetics , Receptor, PAR-1/physiology , Recombinant Fusion Proteins/physiology , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction , Thromboplastin/biosynthesis , Thromboplastin/genetics , Transfection
17.
Zhongguo Zhong Yao Za Zhi ; 30(14): 1102-5, 2005 Jul.
Article in Chinese | MEDLINE | ID: mdl-16161449

ABSTRACT

OBJECTIVE: To explore the effects of xue-bao capsules on injury of radio-or chemo-therapy in mice, in order to provide rationale behind clinical trials. METHOD: xue-xu (deficiency of blood) model in mice was induced by radiation or cyclophosphamide. Leucocyte (WBC), erythrocyte (RBC), hemoglobin (Hb) and platelet (Pt) in peripheral blood as well as CFU-E and CFU-Gm in bone marrow were counted. RESULT: CFU-E and CFU-Gm in normal mice were promoted by this drug. The reduction of WBC, RBC and Hb in peripheral blood as well as CFU-E and CFU-Gm in bone marrow owing to the 3.5 Gy of 60Co radiation were antagonized by the drug. It had also antagonized cyclophosphamide induced the reduction of WBC, RBC and Pt in peripheral blood. CONCLUSION: xue-bao capsules has the effects against the adverse reactions of radio-or-chemo-therapy.


Subject(s)
Bone Marrow Cells , Drugs, Chinese Herbal/pharmacology , Plants, Medicinal , Radiation Injuries, Experimental/pathology , Animals , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Bone Marrow Cells/radiation effects , Capsules , Cell Count , Cells, Cultured , Cyclophosphamide/toxicity , Drug Combinations , Drugs, Chinese Herbal/administration & dosage , Drugs, Chinese Herbal/isolation & purification , Erythrocyte Count , Erythroid Precursor Cells/drug effects , Erythroid Precursor Cells/radiation effects , Female , Granulocyte Precursor Cells/drug effects , Granulocyte Precursor Cells/radiation effects , Leukocyte Count , Male , Mice , Plants, Medicinal/chemistry , Platelet Count , Radiation Injuries, Experimental/blood , Random Allocation , Whole-Body Irradiation/adverse effects
18.
Yi Chuan ; 27(4): 623-8, 2005 Jul.
Article in Chinese | MEDLINE | ID: mdl-16120590

ABSTRACT

In this study, the encoding sequences of SARS-CoV spike protein were analyzed by bioinformatics methods, the structural characteristics and functions were forecasted based on available data. It suggests that the fragment of spike glycoprotein (S401-659) may be crucial for viral attachment and may be a major immunodominant epitope. Then the fragment was amplified and subcloned into expression vector pET28a(+) and pPIC9K. These two plasmids pET28a(+)-S and pPIC9K-S were transformed to E.coli strain BL21(DE3)-star and Pichia pastoris, respectively. SDS-PAGE and Western blot analysis showed that the recombinant protein was successfully expressed. The denatured inclusion bodies were purified with Ni-NTA chelate agarose and its purity can reach 90%.


Subject(s)
Membrane Glycoproteins/metabolism , Recombinant Proteins/metabolism , Severe acute respiratory syndrome-related coronavirus/metabolism , Viral Envelope Proteins/metabolism , Blotting, Western , Epitopes/chemistry , Epitopes/genetics , Epitopes/metabolism , Escherichia coli/genetics , Gene Expression Regulation, Viral , Genetic Vectors/genetics , Membrane Glycoproteins/chemistry , Membrane Glycoproteins/genetics , Pichia/genetics , Protein Sorting Signals/genetics , Protein Structure, Tertiary , Recombinant Proteins/chemistry , Severe acute respiratory syndrome-related coronavirus/genetics , Spike Glycoprotein, Coronavirus , Viral Envelope Proteins/chemistry , Viral Envelope Proteins/genetics
19.
J Biochem ; 135(4): 555-65, 2004 Apr.
Article in English | MEDLINE | ID: mdl-15115782

ABSTRACT

Anti-tumor associated antigen (TAA).CD3.CD28 trispecific antibody(TsAb) is able to provide two signals for fully and continuously activation of T lymphocytes and recruit them around tumor cells, presenting an attractive concept in tumor immunotherapy. Here, a new single chain trispecific antibody (scTsAb), named CEA-scTsAb, was constructed by fusion of anti-CEA (Carcinoma Embryonic Antigen) single chain antibody (scFv), anti-CD3 scFv and anti-CD28 VH, spaced by polypeptide interlinkers taken from the fragment of constant region (FC) of human IgG and human serum albumin (HSA). It was expressed in Escherichia coli at low temperature (30 degrees C) with up to 50% of the antibody being present in soluble form. After one step of DEAE anion chromatography, the soluble product was sufficiently pure for further in vitro activity assays. First, it was proved that CEA-scTsAb could recognize three antigens (CEA, CD28 and Jurkat cell membrane antigen) specifically and could distinguish antigen positive cells from antigen negative cells in vitro. Then fresh PBMC (peripheral blood mononuclear cells), without being pre-treated by co-stimulatory reagents, such as IL-2 or CD28 mAb, were used as effector cells to test their ability to mediate tumor specific cytolysis of CEA-positive tumor cells, SW1116. It was found by photomicrography that T lymphocytes were attracted to SW1116 cells in the presence of CEA-scTsAb, which resulted in effective cytolysis of tumor cells. As shown by MTT assay, the efficacy of tumor specific cytolysis mediated by CEA-scTsAb related to both the quantity and activation of PBMC. At an effector cells/target cells ratio (E/T) of 5, it was proved by dual-color FACS with propidium iodide (PI) and FITC-annexin V that both necrosis and apoptosis of tumor cells were causes of tumor specific cytolysis. In summary, a new single chain trispecific (CEA x CD3 x CD28) antibody was constructed and characterized carefully in this paper and was found to possess functions: (i) to activate T lymphocytes independently of additional co-stimulatory signal, (ii) to attract activated T lymphocytes around CEA-positive tumor cells, (iii) to attack CEA-positive tumor cells with recruited T lymphocytes. Because it recognizes a widely distributed tumor antigen (CEA), with moderate molecular weight (about 75 kDa) and a simple production procedure, and is able to mediate a high level of tumor specific cytolysis without any additional co-stimulating reagents, CEA-scTsAb is very promising for the task of immunotherapy in future.


Subject(s)
Antibodies, Monoclonal/pharmacology , Carcinoembryonic Antigen/immunology , Cytotoxicity, Immunologic/immunology , T-Lymphocytes/immunology , Antibodies, Monoclonal/genetics , Antibodies, Monoclonal/immunology , Apoptosis/immunology , CD28 Antigens/immunology , CD3 Complex/immunology , Cell Line, Tumor , Chromatography, Ion Exchange , Coculture Techniques , Cytotoxicity, Immunologic/drug effects , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , Escherichia coli/genetics , Flow Cytometry , Gene Expression , Genetic Vectors/genetics , Humans , Immunoglobulin Fc Fragments/genetics , Immunoglobulin Fragments/genetics , Immunoglobulin Heavy Chains/genetics , Immunoglobulin Variable Region/genetics , Jurkat Cells , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/immunology , Lymphocyte Activation/immunology , Necrosis/immunology , Polymerase Chain Reaction , Recombinant Fusion Proteins/immunology , Recombinant Fusion Proteins/isolation & purification , Recombinant Fusion Proteins/pharmacology , Serum Albumin/genetics , T-Lymphocytes/drug effects
20.
J Biochem ; 136(6): 769-76, 2004 Dec.
Article in English | MEDLINE | ID: mdl-15671487

ABSTRACT

Secondary lymphoid tissue chemokine (SLC) is a CC chemokine that plays an important role in leukocytes homing to lymphoid tissues. The ability of SLC to co-localize both T cells and dendritic cells formed the rationale to evaluate its utility in cancer immunotherapy. The in vivo antitumor effect of murine SLC (mSLC) has been well documented, but little is known about that of human SLC (hSLC). To investigate the antitumor efficiency in vivo of hSLC, the hSLC gene was artificially synthesized and induced to express as a soluble form in Escherichia coli. After purification, the purity of the recombinant human SLC (rhSLC) protein was above 95% by SDS-PAGE analysis. The K(d) of rhSLC binding to peripheral blood lymphocytes (PBLs) was 0.2186 +/- 0.02675 microM as assessed by FACS, and the maximal chemotactic index of rhSLC was 9.49 at 100 nM as assessed by in vitro chemotaxis assay. Then genomic sequences of hSLC and mSLC, and of human CCR7 (hCCR7) and murine CCR7 (mCCR7), the receptor for SLC, were aligned. It was found that hSLC and mSLC share 70.72% identity and hCCR7 and mCCR7share 86.77% identity. Furthermore, we found that rhSLC could chemoattract murine peripheral blood mononuclear cells (PBMCs) in vitro. On the basis of these facts, immune competent mice inoculated with S180 sarcoma cells were chosen as an in vivo model. Intratumoral injections of rhSLC inhibited tumor growth and increased survival. These findings suggest that, despite its incapability to bind to either human or murine CXCR3, which is related to angiostasis, rhSLC can induce an antitumor response in vivo by another route. This report proves that rhSLC has a potent tumor-inhibition ability that makes it a promising candidate agent in cancer immunotherapy.


Subject(s)
Angiogenesis Inhibitors/immunology , Angiogenesis Inhibitors/therapeutic use , Chemokines, CC/immunology , Chemokines, CC/therapeutic use , Genetic Therapy , Sarcoma, Experimental/prevention & control , Adult , Angiogenesis Inhibitors/metabolism , Animals , Chemokine CCL21 , Chemokines, CC/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Humans , In Vitro Techniques , Lymphocytes/immunology , Lymphocytes/metabolism , Lymphocytes/pathology , Mice , Mice, Inbred BALB C , Mice, Inbred ICR , Mice, SCID , Receptors, Chemokine/metabolism , Recombinant Proteins/therapeutic use , Research , Sarcoma, Experimental/immunology , Sarcoma, Experimental/metabolism , Survival Rate , T-Lymphocytes/immunology , Tumor Cells, Cultured
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