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1.
Article in Chinese | WPRIM | ID: wpr-862604

ABSTRACT

Objective@#To explore the intrinsic mechanism of emotional intelligence and trait anger in the association between childhood psychological abuse and aggressive behavior among middle school students,and to provide a theoretical basis for intervention of aggressive behaviors.@*Methods@#By using the multi-stage stratified random sampling method, 2 458 middle school students in Guizhou Province were selected, and group tests were carried out using psychological abuse scale, attack behavior scale,emotional intelligence scale and characteristic anger scale.@*Results@#Agressive behavior, trait anger and emotional intelligence differed significantly by gender(t=2.19,5.12,-2.34,P<0.05), and there were significant differences in aggressive behavior, psychological abuse, emotional intelligence and trait anger among middle school students of different student origins by residence(t=-18.77,-6.04,9.10,-9.94,P<0.05). Childhood psychological abuse (r=0.41), temperamental trait anger (r=0.52) and reactive trait anger (r=0.49) were significantly positively correlated with aggressive behavior(P<0.05). Emotional intelligence was significantly negatively correlated with aggressive behavior (r=-0.33, P<0.05). The test of multiple mediation effects showed that emotional intelligence, temperamental trait anger and reactive trait anger played multiple mediation roles between psychological abuse and aggressive behavior in childhood.@*Conclusion@#Childhood psychological abuse could not only directly affect the aggressive behavior of middle school students, but also further affect the aggressive behavior through emotional intelligence and trait anger. Emotional intelligence and idiosyncratic anger are important internal mechanisms by which childhood psychological abuse affects aggressive behavior.

2.
Chinese Pharmaceutical Journal ; (24): 930-934, 2016.
Article in Chinese | WPRIM | ID: wpr-859098

ABSTRACT

OBJECTIVE: To determine two toxic impurities, namely bromoacetic acid and 4-aminobenzonitrile, in the intermediate of dabigatran etexilate by UHPLC-MS. METHODS: The separation was performed on a Shimadzu Shim-Pack GIS C18 column (2.1 mm×50 mm, 2 μm) with mobile phase consisting of 0.1% formic acid aqueous solution (A) and 0.1% formic acid methanol (B) by gradient elution at a flow rate of 0.4 mL·min-1. The detection was achieved by triple quadrupole mass spectrometry with rapid polarity switching using MRM mode. RESULTS: The calibration curves were linear in the ranges of 0.2-40 and 0.4-40 ng·mL-1 for bromoacetic acid and 4-aminobenzonitrile, respectively. The values of LOQ of bromoacetic acid and 4-aminobenzonitrile were 0.1 and 0.4 ng·mL-1, respectively. The recoveries of bromoacetic acid and 4-aminobenzonitrile were 100.9% and 99.6%, respectively. CONCLUSION: The method is accurate, rapid, sensitive, and reliable to determine the two toxic impurities bromoacetic acid and 4-aminobenzonitrile in the intermediate of dabigatran etexilate for quality control.

3.
Zhonghua Yu Fang Yi Xue Za Zhi ; (12): 848-854, 2013.
Article in Chinese | WPRIM | ID: wpr-355775

ABSTRACT

<p><b>OBJECTIVE</b>The health surveillance proposal for chromate exposed workers was provided and analyzed on the evidence-based study and then to be improved.</p><p><b>METHOD</b>Firstly, the related literatures were searched about liver damage, micronuclei, urinary chromium and hexavalent chromium exposure in Evidence Based Medicine Reviews such as Cochran library, OVID Medline, Web of knowledge in December 2011; and then, these literatures were reviewed in according to inclusion and exclusion criteria; 22 articles totally were retrieved, evaluated and classified in according to the grading standard by Oxford Centre for Evidence-based Medicine.Finally, field epidemiological investigation was further adopted to confirm the efficiency and feasibility of this proposal, combined with cost-effectiveness analysis:the ratio of total cost divided survival years was used to express the cost-effectiveness.</p><p><b>RESULT</b>Only the glutamic pyruvic transaminase test could not reflect liver damage caused by chromate exposure well; Urinary chromium correlated well with the index reflecting body damage caused by chromate exposure; Binucleated cells micronucleus index in peripheral blood lymphocyte could reflect the genetic damage caused by chromate exposure. As for health economic evaluation of chromate lung cancer, the value of cost/effectiveness was ¥42 321.61 per year that was far below the value of common people (¥252 868.97 per year) .</p><p><b>CONCLUSION</b>It was suggested that serum glutamic pyruvic transaminase test should be replaced by liver function test, urinary chromium should be classified as a compulsory index and binucleated cells micronucleus index in peripheral blood lymphocyte should be supplied as a recommended index.</p>


Subject(s)
Humans , Alanine Transaminase , Blood , Chromates , Urine , Evidence-Based Medicine , Micronucleus Tests , Occupational Exposure , Population Surveillance
4.
Zhongguo Zhong Yao Za Zhi ; (24): 3628-3632, 2012.
Article in Chinese | WPRIM | ID: wpr-346894

ABSTRACT

<p><b>OBJECTIVE</b>To do some comparative study on anti-inflammatory and antipyretic effects between the Dao-di herb and non Dao-di herb of Huangqin (the roots of Scutellaria baicalensis) and provide thinking and evidence for study on geoherbalism and clinical usage of Huangqin.</p><p><b>METHOD</b>The anti-inflammatory action was assessed by auricular swelling induced by dimethylbenzene in mice and the antipyretic action was monitored by dried yeast-induced mice fever.</p><p><b>RESULT</b>All samples of both Dao-di and non Dao-di herbs of Huangqin showed antipyretic effect. The Dao-di Huangqin samples showed antipyretic effect between 61% to 53% , whereas the non Dao-di Huangqin samples between 53% to 43%. Six Dao-di Huangqin samples showed better antipyretic effect than four non Dao-di Huangqin samples. All samples of both Dao-di and non Dao-di Huangqin showed anti-inflammatory effect. Dao-di Huangqin showed anti-inflammatory effect between 73% to 54%, whereas non dao-di Huangqin between 53% to 34%. Six Dao-di Huangqin showed better anti-inflammatory effect than four non Dao-di Huangqin. In totality, results from analysis of geoherbalism showed that geoherbal production areas of Huangqin had better effect than that of the non geoherbal production areas in anti-inflammatory and antipyretic effects.</p><p><b>CONCLUSION</b>Both the Dao-di and non Dao-di Huangqin have effects of anti-inflammatory and antipyretic to a certain extent, but the efficacy of the Dao-di Huangqin surpass the non Dao-di Huangqin.</p>


Subject(s)
Animals , Humans , Mice , Anti-Inflammatory Agents , Antipyretics , China , Drug Contamination , Drugs, Chinese Herbal , Fever , Drug Therapy , Inflammation , Drug Therapy , Scutellaria baicalensis , Chemistry
5.
Zhonghua Yu Fang Yi Xue Za Zhi ; (12): 355-358, 2012.
Article in Chinese | WPRIM | ID: wpr-292467

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of combined occupational exposure of chromium and iron on erythrocyte metabolism, and the possible mechanism.</p><p><b>METHODS</b>A total of 115 chromate production workers were selected in a chemical factory of Jinan as exposure group, Dec, 2008, and 60 healthy residents from a community which was far away from the factory were enrolled as control group. Environmental concentrations of chromium and iron were collected by filter membrane sampling and determined. The peripheral blood of subjects were collected for determination of chromium, iron, copper in whole blood and folate, vitamin B₁₂ in serum, mean corpuscular hemoglobin (MCH) and mean corpuscular volume (MCV) and correlation analysis was conducted.</p><p><b>RESULTS</b>The median (quartile interval) concentration of air-chromium and air-iron in workplace were 9.0 (10.5) and 11.2 (10.1) µg/m³, respectively, which were significantly higher than that of the control (0.1 (0.1) and 7.2 (2.5) µg/m³) (all P values < 0.01). Blood-chromium and blood-iron of the exposed group were 15.5 (14.1) µg/L and (895.1 ± 90.2) mg/L, which were significantly higher than the counterpart of the control (3.6(2.0) µg/L, (563.7 ± 49.3) mg/L) (all P values < 0.01). Serum folate ((6.9 ± 2.5) µg/L), serum vitamin B₁₂ ((396.4 ± 177.0) µg/L) and blood copper ((777.6 ± 103.5) µg/L) of the exposed group were all significantly lower comparing to the control group ((558.0 ± 330.8), (8.1 ± 3.8), (812.1 ± 94.6) µg/L) (all P values < 0.05). The relationships between blood chromium and serum folate, serum vitamin B₁₂ were statistical significant (r = -0.319 and -0.293, P < 0.01). Both serum vitamin B₁₂ and blood copper correlated with mean corpuscular hemoglobin (MCH) and mean corpuscular volume (MCV) (r = -0.223, -0.242, -0.261, -0.292, all P values < 0.01).</p><p><b>CONCLUSION</b>Combined chromium and iron exposure existed in the workplace. Adverse effect of Chromium on human erythrocyte may via folate and vitamin B₁₂ metabolism, while iron may via copper metabolism.</p>


Subject(s)
Humans , Air Pollutants, Occupational , Chromates , Chromium , Copper , Blood , Erythrocytes , Metabolism , Folic Acid , Blood , Iron , Occupational Exposure , Vitamin B 12 , Blood
6.
Zhonghua Yu Fang Yi Xue Za Zhi ; (12): 775-779, 2010.
Article in Chinese | WPRIM | ID: wpr-291479

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the inhalable titanium dioxide exposure level and make an assessment of its oxidative effect on occupational exposed population.</p><p><b>METHODS</b>A total of 7 workers occupationally exposing to inhalable titanium dioxide were recruited into the study. The basic information and occupational history were collected by interview, while their blood sample (10 ml for each subject) were collected before and after the investigation, respectively. Pre- and post-work shift urine samples (60 ml for each subject) were collected for 29 days consecutively. The daily personal titanium dioxide exposure level, temperature and relative humidity were detected too. Urinary 8-hydroxy-deoxyguanosine (8-OHdG) and serum high-sensitivity C-reactive protein (hs-CRP) were detected by ELISA and latex immunoturbidimetric assay, respectively.</p><p><b>RESULTS</b>The mean concentration of air inhalable titanium dioxide was (1.194 ± 1.015) mg/m(3). Serum hs-CRP level before and after the investigation was (1.13 ± 1.08), (1.33 ± 1.01) mg/L, respectively. No statistical significance was observed between hs-CRP level before and after the investigation (t = -0.848, P = 0.425). Pre- and post-work shift urinary 8-OHdG was (3.51 ± 1.39), (3.65 ± 1.06) µmol/mol Cr, respectively. A positive correlation was found between the concentration of inhalable titanium dioxide and the changes of 8-OHdG level (r = 0.192, t = 2.09, P = 0.039). Linear mixed-effect models, adjusted by work shift, years of employment, age, body mass index, smoking status, temperature and relative humidity, showed no significant exposure-respond trend between the inhalable titanium dioxide concentration and 8-OHdG level (β = 0.288, t = 1.940, P = 0.055).</p><p><b>CONCLUSION</b>Our findings do not support the potential link between occupationally exposure to inhalable titanium dioxide and high induction of DNA oxidative stress.</p>


Subject(s)
Humans , Male , Middle Aged , Air Pollutants, Occupational , C-Reactive Protein , Deoxyguanosine , Urine , Occupational Exposure , Oxidative Stress , Titanium , Blood , Urine
7.
Article in Chinese | WPRIM | ID: wpr-237617

ABSTRACT

The aim of this study was to detect the presence of human AML leukemia stem cells (LSC) in childhood patients with acute leukemia (AL) and analyze the correlation between LSC concentrations and minimal residual disease (MRD) levels in AML cases after remission. The multi-parameter flow cytometry (FCM) and a panel of monoclonal antibody combination were used to detect the AML LSC or AML LSC immunophenotype-identical cell (AML LSC-IPIC) concentrations in childhood AML or ALL leukemia both at new diagnosis and at remission and correlated AML LSC to the MRD levels at different time points after remission. The results indicated that the AML LSC or AML LSC-IPIC concentrations [in average 166 (range 14 - 1459)/100 000 mononuclear cells (MNCs)] in AML at initial diagnosis were significantly higher than those in ALL [7 (range 0 - 560)/100 000 MNCs, p < 0.017] and control [0 (range 0 - 6)/100 000 MNCs, p < 0.017], respectively. The AML LSC concentrations in AML at non-CR were in average 36 (range 5 - 224)/100 000 MNCs. No statistical difference (p > 0.05) was found between the AML LSC or AML LSC-IPIC concentrations in AML (in average 6 (range 0 - 41)/100, 000 MNCs) and ALL [10 (range 0 - 105)/100, 000 MNCs] after CR. The significantly negative correlation was noticed between AML LSC concentrations and MRD levels. It is concluded that the AML LSCs exist in newly diagnosed AML, which are significantly reduced when complete remission has achieved, but the low levels of these populations still remain. The phenotypically similar (CD34(+)CD38⁻CD123(+)) AML LSC populations have also been found in the bone marrow from ALL patients, but their concentrations are not significantly different when CR has achieved. The significantly negative correlation between AML LSC concentrations and MRD levels is observed in AML patients after remission.


Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , Flow Cytometry , Immunophenotyping , Leukemia, Myeloid, Acute , Allergy and Immunology , Metabolism , Neoplasm, Residual , Allergy and Immunology , Metabolism , Neoplastic Stem Cells , Cell Biology , Tumor Stem Cell Assay
8.
Chinese Journal of Pediatrics ; (12): 57-61, 2009.
Article in Chinese | WPRIM | ID: wpr-306961

ABSTRACT

<p><b>OBJECTIVE</b>Leukemia is the most common hematopoietic malignancies in children. Chemotherapy is currently the primary modality of treatment for this fatal disease. Although chemotherapy is very effective in terms of cell killing, severe side effects such as severe infections, intracranial hemorrhage etc. are frequently encountered due to its poor selective damage between normal and malignant cells or tissues. Thus, a new therapy with highly selective killing of malignant cells which leaves the normal cells unaffected is desperately desired. The aim of this study was to investigate the targeting efficacy in vitro with a new clone of anti-human CD19 antibody immunotoxin 2E8-Genistein on B lineage leukemia cell line Nalm-6 cells and its mechanisms in order to provide the evidence of target therapy on B lineage leukemia and lymphoma.</p><p><b>METHODS</b>2E8-Genistein immunotoxin was generated by conjugating Mab 2E8 with a tyrosine kinase inhibitor, Genistein (Gen) via the Sulfo-SANPAH, an ultra-violet sensitive reagent. Nalm-6, a CD19+ B cell leukemia cell line, was used as target cells, while Molt-3, a CD19-T cell leukemia cell line, was taken as the negative control. The morphology of the cells was observed under the reverted reversed light microscope and the viability was checked with either trypan blue exclusion or MTT methods. Two-color flow cytometry was applied to study the mechanism of cell killing.</p><p><b>RESULTS</b>After 24 hours of culture, 2E8-Genistein showed marked target killing on Nalm-6 cells at nine different concentrations from 20 nmol/L through 100 nmol/L with cell survival rates from (71.8 +/- 7.9)% down to (16.6 +/- 12.9)%, respectively (n = 3), which were all significantly lower than that of control group (100 +/- 13.9)% (P < 0.05). The killing effect was even more significant when the concentration was over 80 nmol/L. The growth inhibition rates of this immunotoxin on Nalm-6 cells were 82%, 84% and 94%, respectively at 24, 48 and 72 hours of culture in a time dependent manner. Significant difference was observed between the cell growth curve of Nalm-6 cultured with 100 nmol/L of 2E8-Gen and those of Nalm-6 cultured with medium (blank), PBS (negative control) or the same concentration of pure 2E8 antibody (negative control) groups (F = 152.15, P = 2.15 x 10(-7)), but there was no significant difference among the three control groups (F = 1.51, P = 0.29). When Molt-3 cells were used as target cells, the cell growth curves of Molt-3 cultured with 2E8-Gen (100 nmol/L) and with negative control of blank did not show any significant difference (F = 0.34, P = 0.59). PI/FITC Annexin V double staining analysis with flow cytometry showed that the positive rate (33.45 +/- 8.77)% of early apoptosis on Nalm-6 cells induced by 100 nmol/L of 2E8-Genistein was significantly higher than that of negative control of blank (10.44% +/- 1.28%, t = -4.39, P = 0.001) at 24 hours of culture.</p><p><b>CONCLUSION</b>2E8-Genistein immunotoxin can significantly target the Nalm-6 cells in vitro in a time response manner and the apoptosis induction is involved in the course of this killing effect.</p>


Subject(s)
Humans , Antibodies, Monoclonal , Allergy and Immunology , Pharmacology , Antigens, CD19 , Apoptosis , Cell Line, Tumor , Flow Cytometry , Genistein , Allergy and Immunology , Pharmacology , Immunotoxins , Allergy and Immunology , Pharmacology , Leukemia, B-Cell , Allergy and Immunology
9.
Chinese Journal of Pediatrics ; (12): 605-609, 2008.
Article in Chinese | WPRIM | ID: wpr-300723

ABSTRACT

<p><b>OBJECTIVE</b>Acute monocytic leukemia (AML)-M5 is the common type of acute myeloid leukemias in children. Studies have shown that there are abundant lipopolysaccharide (LPS) receptor (designated as CD14) molecules on the cell membrane of M5 cells and they play an important role in the diagnosis of M5, since they can be recognized and bound by mouse-anti-human CD14 monoclonal antibody (McAb). However, mouse-originated antibodies are largely not suitable for clinical application due to the severe side effects, thus "humanized antibody" is desired. As the first step for developing humanized antibody, the construction and expression of single chain antibody (scFv) with functional protein are necessary. The present study aimed to express ZCH-7-2F9 ScFv (scFv(2F9)) in eukaryotic cells and obtain the scFv(2F9) protein with a high biological activity.</p><p><b>METHODS</b>Four primers were synthesized to construct the eukaryotic expressional vector, which included SfiI and EcoRI enzyme cleaving site, 6 x His and stop code TGA sequences. scFv(2F9) gene was amplified through splicing by overlap extension (SOE) using the high fidelity Taq polymerase. Positive recombinants (pSectag2A/scFv(2F9)) were identified through enzyme cleaving and sequenced before expression and were transformed into Chinese hamster ovary (CHO) cells for expression. Western-Blot and flow cytometry (FCM) were carried out to determine the relative molecular mass (Mr) and binding activity of scFv(2F9).</p><p><b>RESULTS</b>The cloned scFv(2F9) gene was identified to be functional by sequencing and expressing. The interested protein could be detected in the culture supernatant of transformed CHO cells with an Mr of 31 000. The blocking test showed that the positive cell percentages, the mean fluorescence intensity (MFI) and the peak of channel (peak Ch) were reduced by 90.02%, 63.30% and 63.38%, respectively after blocking with scFv(2F9), while those were 4.55%, 10.09% and 5.02% after blockage using the supernatant from the CHO cells transfected with blanked vector pSectag2A.</p><p><b>CONCLUSIONS</b>The scFv(2F9) against human CD14 antigen was successfully expressed in eukaryotic cells and showed a high biological activity, which may be useful for the further studies on its humanized antibodies.</p>


Subject(s)
Animals , Child , Cricetinae , Humans , Amino Acid Sequence , Antibodies, Monoclonal , Genetics , Base Sequence , CHO Cells , Cloning, Molecular , Cricetulus , Lipopolysaccharide Receptors , Allergy and Immunology , Molecular Sequence Data , Single-Chain Antibodies , Genetics
10.
Chinese Journal of Pediatrics ; (12): 493-497, 2008.
Article in Chinese | WPRIM | ID: wpr-326101

ABSTRACT

<p><b>OBJECTIVE</b>Monoclonal antibody (mAb) conjugated with certain toxin to generate immunotoxin bears an important and promising effect as a new therapy for patients with hematopoietic malignancies. However, most toxic moieties conjugated with antibody proteins reported in the literature were toxic proteins which presented immunogenicity to patients capable of inducing anti-toxin antibody. Norcantharidin (NCTD) is a small molecule toxin. It does not have the immunogenicity to human body so that it bears a promising potential for development of new targeting drug. In this study, a new clone of self-made anti-CD19 mAb named ZCH-4-2E8 conjugated with NCTD was used to investigate its targeting efficacy against CD19+ lymphoid malignant Nalm-6 cells in vitro to provide the experimental data for the further development of this new targeting agent.</p><p><b>METHODS</b>A monoclonal antibody named 2E8 was prepared from mouse ascites and purified by gel chromatography. The purity of the antibody protein was checked by SDS-PAGE assay. Immunotoxin 2E8-NCTD was successfully generated through conjugating CD19 mAb protein and Norcantharidin by the activated ester method. The binding activity of the immunoconjugate (2E8-NCTD) against CD19 antigens on cell surface and the expression levels of CD19 antigens on Nalm-6 and K562 cells were examined by flow cytometry. Comparisons of the inhibitory effects among PBS, purified 2E8 antibody, norcantharidin and immunotoxin 2E8-NCTD groups on cell growth of either Nalm-6 cells or K562 cells were made.</p><p><b>RESULTS</b>The purity of the purified 2E8 antibody was higher than 99.00% demonstrated by SDS-PAGE assay. 2E8 antibody in the supernatant reacted with 99.34% of Nalm-6 cells, while only 0.98% of K562 cells reacted with this antibody. The newly generated immunotoxin (2E8-NCTD) had a positive rate of 99.90% on Nalm-6 cells with little reduction of binding activity. From the in vitro study, both 2E8-NCTD and norcantharidin were shown to have significant inhibitory effects on the growth of CD19+ Nalm-6 cells (P < 0.001), while the purified 2E8 antibody did not show any significant influences on the growth of Nalm-6 cells. Since no significant inhibitory effects were identified among immunotoxin 2E8-NCTD, 2E8 antibody and control groups on CD19(-) K562 cells, a significant targeting effect of the 2E8-NCTD against Nalm-6 cells was confirmed.</p><p><b>CONCLUSIONS</b>The immunotoxin 2E8-NCTD was successfully synthesized by activated ester method with an excellent targeting killing effect on CD19+ Nalm-6 leukemia cells in vitro, which provides some experimental data for the further development of this new targeting agent.</p>


Subject(s)
Animals , Humans , Mice , Antibodies, Monoclonal , Antigens, CD19 , Allergy and Immunology , Bridged Bicyclo Compounds, Heterocyclic , Allergy and Immunology , Electrophoresis, Polyacrylamide Gel , Hybridomas , Immunotoxins , Allergy and Immunology , K562 Cells , Mice, Inbred BALB C
11.
Article in Chinese | WPRIM | ID: wpr-310421

ABSTRACT

<p><b>OBJECTIVE</b>To establish an acute leukemia animal model for testing new therapeutic agents in vivo.</p><p><b>METHODS</b>Nude mice were intraperitoneally injected with 2 mg cyclophosphamide, 24 h later 5 x 10(6) acute B-cell leukemia Nalm-6 cells was inoculated via the tail vein, then monitored daily. When animals were paralyzed or dying, the organs including the liver, spleen, lung, heart, kidney, brain, bone marrow, pancreas, testes were removed and fixed with formalin, examined by routine histopathology.</p><p><b>RESULTS</b>After Nalm-6 cells were inoculated the mean survival of mice were( 19.4+/-0.55)d (n=6). The paralysis of mice was followed by weight loss, bent spines, hogback, cachexia and death. Histopathological examination showed that the tumor cells infiltrated liver, spleen, kidney, lung, meninges, interior cerebrum, the liver and kidney were the most affected organs.</p><p><b>CONCLUSION</b>B lineage acute leukemia animal model has been successfully established in the nude mice, which is suitable for testing new therapeutic agents.</p>


Subject(s)
Animals , Female , Mice , Cyclophosphamide , Disease Models, Animal , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation , Precursor Cell Lymphoblastic Leukemia-Lymphoma
12.
Article in Chinese | WPRIM | ID: wpr-360516

ABSTRACT

The pharmacokinetic research of Chinese drugs is still in the exploratory stage so far, a great progress has achieved in the researches on active components of them since the 1980s. The progresses in pharmacokinetic research of active components of some commonly used Chinese drugs were reviewed in this paper, and the problems to be solved in future were pointed out.


Subject(s)
Animals , Humans , Biomedical Research , Drugs, Chinese Herbal , Pharmacokinetics , Medicine, Chinese Traditional
13.
Article in Chinese | WPRIM | ID: wpr-344377

ABSTRACT

<p><b>OBJECTIVE</b>To construct a prokaryotic vector of ZCH-7-2F9 single chain antibody (ScFv2F9) and to obtain the ScFv2F9 protein with biological activity for further studies.</p><p><b>METHODS</b>Primers were synthesized according to the gene sequence of ScFv2F9, four tandem glycin and one serine (G4S) 3linker and multiple cloning site(MCS) of pIVEX2.3-MCS vector, which included NdeI and SmaI enzyme cleaving sites. ScFv2F9 gene was amplified through splicing by overlap extension (SOE) using the high fidelity Taq polymerase. Then the gene was cloned to pGEM-T easy and pIVEX2.3-MCS vectors. Positive recombinants (pIVEX2.3-MCS/ScFv2F9) were identified through enzyme cleaving and sequenced before expression. The recombinant plasmids were transfected into E.coli BL21star(DE3)plysS for expression. After purification with Ni+resin and renaturation in vitro, the relative molecular mass (Mr) and the binding activity of the interesting protein were determined by SDS-PAGE and flow cytometry (FCM), respectively.</p><p><b>RESULT</b>The cloned ScFv2F9 gene was identified to be functional by sequencing and expressing. The interesting protein was detected in inclusion body with a Mr of 31 000. The blocking test showed that the positive cell percentage, the mean fluorescence intensity (MFI) and the peak of channel (peak Ch) were reduced by 11.73%, 11.96% and 31.57%, respectively after once blockage by scFv2F9 protein, and 26.44 %, 21.75 % and 42.11 % after blockage twice.</p><p><b>CONCLUSION</b>The ScFv2F9 against human CD14 antigen has been successfully expressed in prokaryotic cells with partial biological activity, which lays the foundation for further studies on its immunotoxin and other kinds of engineered antibodies.</p>


Subject(s)
Humans , Antibodies, Monoclonal , Biotechnology , Methods , Cells, Cultured , Cloning, Molecular , Gene Expression , Genetic Vectors , Lipopolysaccharide Receptors , Allergy and Immunology , Prokaryotic Cells , Metabolism , Recombinant Fusion Proteins , Genetics
14.
Article in Chinese | WPRIM | ID: wpr-269104

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effect of Danzhi Xiaoyao Powder (DXP) on neuro-immuno-endocrine system in patients with depression.</p><p><b>METHODS</b>A randomized double-blinded and controlled study was conducted in 63 cases of depression. They were assigned to the DXP group (32 cases, treated with DXP) and the control group (31 cases, treated with maprotiline). The curative effect was evaluated with Hamilton's depressive scale (HAMD) before and at the end of the 2nd, 4th and 6th week of the treatment. Serum levels of serotonin (5-HT), norepinephrine (NE), brain derived neurotrophic factor (BDNF), cortisol (CORT), interleukin-6 (IL-6), and interleukin-1beta (IL-1beta) were detected before and at the 6th week of the treatment.</p><p><b>RESULTS</b>After 2 weeks of treatment, the total score of HAMD decreased remarkably in both groups (P < 0.01), and the total score, as well as the scores of the three factors, i. e. anxiety/somatization, cognitive impairment and feeling of despair, were lower in the DXP group than that in the control group respectively (P < 0.05 or P < 0.01). After 4 and 6 weeks of treatment the total score and score of the three factors all reduced significantly in both groups (P < 0.01), with insignificant difference between the groups. After 6 weeks of treatment, the serum levels of 5-HT and BDNF increased (P < 0.01), and the serum IL-6 level decreased in both groups (P < 0.05 or P < 0.01), the serum CORT level reduced in the DXP group (P < 0.01), while the serum NE level elevated in the control group (P < 0.01).</p><p><b>CONCLUSION</b>DXP is effective in improving symptoms of depression by regulating the levels of 5-HT, BDNF, CORT and IL-6.</p>


Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Brain-Derived Neurotrophic Factor , Blood , Depressive Disorder , Blood , Drug Therapy , Psychology , Double-Blind Method , Drugs, Chinese Herbal , Therapeutic Uses , Interleukin-6 , Blood , Norepinephrine , Blood , Phytotherapy , Psychiatric Status Rating Scales , Serotonin , Blood
15.
Chinese Journal of Pediatrics ; (12): 736-741, 2007.
Article in Chinese | WPRIM | ID: wpr-311733

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the safety and efficacy of sublingual immunotherapy with 'Dermatophagoides Farinae Drops' in D. farinae allergic asthma and/or rhinitis patients.</p><p><b>METHODS</b>A 25-week double-blind, placebo-controlled, multi-centered trail was conducted in 278 children (aged 4 - 18 yr) with mite-induced asthma and/or rhinitis. Patients were randomly assigned to receive sublingual immunotherapy (SLIT) with 'Dermatophagoides Farinae Drops' (n = 139) or placebo (n = 139) for 25 weeks and the dosage and administration strictly followed the manufacturer's instructions. At the beginning of the 2nd, 3rd, 4th, 6th, 10th, 14th, 18th, 22nd week of the treatment, the patients were asked to accept follow-up visit, during the clinical trial all patients and parents were asked to keep a daily record of their asthma symptom scores, rescue medicine use, rhinitis symptom scores, morning and evening peak expiratory flow. Asthma symptom scores, reduction in use of rescue medicine, rhinitis symptom scores, lung function tests, skin sensitivity to mite, mite-specific immunoglobulin (Ig) E and IgG4, and quality of life and adverse effect were assessed during the study.</p><p><b>RESULT</b>(1) Of the 278 children, 27 dropped out before the study completion. (2) After 25 weeks of treatment, the median variability of PEFR was -1.38 for SLIT group and -0.90 for the placebo (P < 0.05). (3) Besides, the mean variability of medicine score of asthma was -0.08 for SLIT group and 0.52 for the plcebo (P < 0.05). (4) The median variability of rhinitis symptom score was -1.96 for SLIT group and -1.03 for the placebo (P < 0.01). (5) The rescue medicine usage of SLIT reduced but did not show significant differences between SLIT and placebo. (6) After 25 weeks treatment, the increase of D. farinae specific IgE antibody of two groups were similar, while specific IgG4 increased significantly in SLIT compared to the patients in control one (P < 0.01); (7) No severe adverse events happened in the trial and the most-likely adverse events were mild asthma and local rash.</p><p><b>CONCLUSION</b>Dermatophagoides Farinae Drops is safe and effective in treating allergic asthma and atopic rhinitis.</p>


Subject(s)
Adolescent , Animals , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Asthma , Therapeutics , Dermatophagoides farinae , Chemistry , Metabolism , Double-Blind Method , Mites , Chemistry , Rhinitis , Therapeutics
16.
Chinese Journal of Pediatrics ; (12): 334-338, 2007.
Article in Chinese | WPRIM | ID: wpr-356184

ABSTRACT

<p><b>OBJECTIVE</b>Leukemia is the most common malignancy in children. Combined chemotherapy is currently the primary treatment modality. During the past decade, very high cure rates of childhood acute lymphoblastic leukemia (ALL) have been reported both at home and abroad. However, the cure rates of children with acute myeloid leukemia (AML) remain low due to the multiple-drug resistance (MDR). P-glycoprotein (P-gp) is one of the most important mechanisms of MDR for leukemia cells. However, the function of the protein, the clinical application of its reversal agents and the efficacy of the combination of the reversal agents remain to be elucidated. The present study aimed to evaluate the P-gp pump function on leukemia cell membrane and the effects of the combined administration of the reversal agents cyclosporin A (CSA) and verapamil (VER) through the observation of Calcein-AM (C-AM) metabolism in the cell line K562 and its multi-drug resistant subline K562/VCR.</p><p><b>METHODS</b>The mean fluorescence intensity (MFI) of C-AM inside the cytoplasm was analyzed with flow cytometry (FCM). The events of K562 and K562/VCR cells treated and untreated with CSA, VER and CSA + VER were acquired at time points 0, 30, 60, 90 and 120 minutes, respectively, and the data obtained were analyzed with CellQuest software.</p><p><b>RESULTS</b>The C-AM in the K562 and K562/VCR varied more apparently in the fist 24 hours. In addition, the MFI of the C-AM in K562 was significantly higher than that in K562/VCR cells indicating that the P-gp pump molecules were functioning. The MFIs of the CSA, VER and CSA + VER groups co-cultured with K562/VCR cells were 4014 +/- 219, 3879 +/- 116 and 4158 +/- 302, respectively after 120 min of incubation, significantly higher as compared to that of control group (3251 +/- 107, P < 0.05). On the other hand, significant inhibition of the efflux from the K562/VCR cell line was also noticed after the same time period of incubation with the MFIs of 2237 +/- 155, 1932 +/- 233 and 2231 +/- 147, respectively in the three groups, which was significantly higher than that of control group (1622 +/- 191, P < 0.05). CSA, VER and CSA + VER could increase the uptake and inhibit the efflux of C-AM by K562/VCR cells, while no evident influence on those functions inside the parental cell line K562 cells was noticed.</p><p><b>CONCLUSIONS</b>CSA, VER and CSA + VER could increase the uptake and reduce the efflux of C-AM by K562/VCR cells while no significant difference between the CSA + VER and CSA or VER was noticed. P-gp pump function and the effects of its reversal agents on leukemic cells can be rapidly and easily evaluated by using the C-AM and FCM.</p>


Subject(s)
Child , Humans , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Metabolism , Antineoplastic Agents , Pharmacology , Drug Resistance, Multiple , Flow Cytometry , Methods , Fluoresceins , Pharmacology , K562 Cells , Tumor Cells, Cultured , Verapamil , Pharmacology , Vincristine , Pharmacology
17.
Zhongguo dangdai erke zazhi ; Zhongguo dangdai erke zazhi;(12): 28-33, 2007.
Article in Chinese | WPRIM | ID: wpr-357756

ABSTRACT

<p><b>OBJECTIVE</b>Acute promyelocytic leukemia (APL) is a specific type of hematopoietic malignancy, accounting for 10% of the de novo acute myeloid leukemia (AML). The data on long-term outcome of APL in children are limited. The aim of this study was to investigate the clinical biological features, diagnosis, prognosis and long-term survival of childhood APL.</p><p><b>METHODS</b>A total of 46 children with newly diagnosed APL from April 1998 to October 2005 were enrolled into this study. Induction treatment containing all-trans retinoic acid (ATRA) plus daunorubicin (DNR) or pirarubicin (THP) was performed on these patients, followed by 6 courses of chemotherapy consolidation: DNR, homoharringtonine or etoposide plus Ara-C. A maintenance therapy was then administered once 3-6 months. The total period of treatment was 2.5 years.</p><p><b>RESULTS</b>Of the 39 patients who had completed the regular treatment, 36 (92.3%) achieved a complete remission. The 5-year cumulative incidence of relapse (CIR) was 28.6%. The estimated overall survival (OS) rates at 1, 3 and 5 years were (86.1 +/- 5.8)%, (76.1 +/- 7.5)% and (70.2 +/- 8.9)% respectively, while the event free survival (EFS) rates were (78.4 +/- 6.8)%, (63.6 +/- 8.7)% and (53.1 +/- 10.0)% respectively. The 5-year OS rate of patients with WBC less than or equal to 10.0 X 10(9)/L was (81.4 +/- 10.3)%, which was significantly higher than that with WBC greater than 10.0 X 10(9)/L[(51.6 +/- 14.7)%, P < 0.05]. Five patients with RT-PCR positive for PML/RARalpha S (short) subtype died eventually although all of them achieved CR, but none of the 13 patients with PML/RARalpha L (long) subtype died.</p><p><b>CONCLUSIONS</b>Remission induction therapy with ATRA + DNR or THP is effective and safe for newly diagnosed childhood APL. The remission induction therapy combined with chemotherapy containing high/intermediate dose Ara-C can improve the long-term survival rates of APL patients. High WBC count and S subtype of PML-RARa are two poor prognostic factors for children with APL.</p>


Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Infant , Male , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Follow-Up Studies , Leukemia, Promyelocytic, Acute , Drug Therapy , Mortality , Oncogene Proteins, Fusion , Genetics , Prognosis , Survival Rate , Treatment Outcome , Tretinoin
18.
Article in Chinese | WPRIM | ID: wpr-282747

ABSTRACT

ZCH-2B8a (IgG2a) is a novel monoclonal antibody (McAb) generated in laboratory of Children Hospital of Medical College, Zhejiang University recently using human myeloblastic leukemia cell line KG1a as immunogen. This antibody has been submitted to the 8th International Workshop and Conference on Human Leukocyte Differentiation Antigens (HLDA8) and the results showed that the antibody recognized an unknown molecule on the surface of some blood cells. The aim of this study was to investigate the reactivity of this antibody on normal blood cells and malignant cell lines and to explore its possible application in clinical practice. The multi-parameter flow cytometry was used to analyze the expression pattern of 2B8a antigen in triplicate on normal blood components including T cells, B cells, natural killers (NK), neutrophils, monocytes, dendritic cells (DC), red blood cells (RBC), platelets (Plt), hematopoietic stem/progenitor cells derived from either bone marrow or G-CSF mobilized peripheral blood CD34(+) cells and malignant cell lines including 14 hematopoietic, 5 neuroblastoma, 1 colon cancer and 1 amniotic epithelium cell lines. The amount of positive cells > or = 20% was considered as positivity. The results showed that 2B8a antibody reacted to 3/3 specimens of blood B cells with a positive rate of 26.29% and 2/3 specimens of monocytes with an average positive rate of 59.84%. 2B8a was weakly reactive to neutrophils (23.72%) and negative for T cells, NK, DC, RBC and Plt. The antibody reacted to all 3 marrow CD34(+) cells with an average positive rate of 39.33% while it was negative for G-CSF-mobilized CD34(+) peripheral blood stem/progenitor cells (PBSC, 1.25%). Cell line analysis showed that the antibody notably reacted to three out of 4 cell lines (Raji, SMS-SB, Nalm-6 and Nall-1) with the positive rates of 98.78%, 98.61%, 94.93% respectively and weakly to one of them with 5.68% in B lineage cell lines and monoblastic cell line (U937, 67.78%) while it was only weakly positive or negative for other myeloid leukemia cell lines including Meg01 (33.40%), HL-60 (29.70%), K562 (28.19%), KG1a (16.23%) and HEL92.1.7 (8.02%). Among 4 T lineage leukemia, 5 neuroblastoma and 1 colon cancer cell lines tested, only Molt-3 was found weakly positive (31.40%) for 2B8a, while the remaining 3 T cell lines (Molt4, JM and CCRF-CEM), 5 neuroblastoma cell lines (LA-N1, KCNR, BE, SK-N-SH, SK-N-AS) and the colon cancer cell line (HR8348) tested were negative. An amniotic epithelium cell line (FL) was showed positive for the antibody (45.03%). It is concluded that 2B8a antibody primarily reacts to B lineage and monocytic lineage cells which may bear the diagnostic and therapeutic applications among different types of hematopoietic malignancies.


Subject(s)
Humans , Antibodies, Monoclonal , Allergy and Immunology , Antibody Specificity , Antigen-Antibody Reactions , Antigens, Neoplasm , Allergy and Immunology , Antigens, Surface , Allergy and Immunology , B-Lymphocytes , Cell Biology , Allergy and Immunology , HLA Antigens , Allergy and Immunology , Hematopoietic Stem Cells , Cell Biology , Allergy and Immunology , Hematopoietic System , Cell Biology , Leukemia, Myeloid, Acute , Allergy and Immunology , Pathology , Tumor Cells, Cultured
19.
Article in Chinese | WPRIM | ID: wpr-331719

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effect and side effect of Danzhi Xiaoyao powder (DXP) in treating depression.</p><p><b>METHODS</b>A randomized controlled and double-blinded study was conducted in 63 cases of depression by divided them into the western medicine group (WMG, 31 cases) treated with maprotiline, and the Chinese medicine group (CMG, 32 cases) treated with DXP. The effect of therapy was evaluated before and at the 2nd, 4th and 6th week of the treatment with Hamilton's depressive scale (HAMD), self-rating depression scale (SDS), self-rating anxiety scale (SAS) and the scale for TCM syndrome and symptom differentiation (TCM-SSD), and the side-effect of therapy was assessed with Asberg side-effect scale as well.</p><p><b>RESULTS</b>There was no significant difference between the two groups in scores of HAMD, SDS, SAS, and TCM-SSD. The markedly effective rate in CMG was 84% and in WMG 87%, showed no significance between them (P > 0.05). The scores of HAMD, SDS and SAS of both groups were remarkably lowered after therapy (P < 0.05). However, the score of Asberg in CMG was lower than that in WMG (P < 0.05).</p><p><b>CONCLUSION</b>DXP shows the effect equivalent to that of maprotiline, but with obviously less side-effect.</p>


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Antidepressive Agents , Therapeutic Uses , Depressive Disorder , Drug Therapy , Double-Blind Method , Drugs, Chinese Herbal , Therapeutic Uses , Maprotiline , Therapeutic Uses , Phytotherapy , Psychiatric Status Rating Scales
20.
Article in Chinese | WPRIM | ID: wpr-332137

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the reactive pattern and its clinical significance of ZCH-7-2D3 monoclonal antibody on leukemia cells.</p><p><b>METHODS</b>Bone marrow samples from 100 leukemia patients (male 59: female 41, 34 cases of children and 66 adults, aged 11 months - 77 year) were collected. A CD45 gating strategy and multi-parameter flow cytometry were used to analyze the leukemia cells.</p><p><b>RESULT</b>The positive rate (10/31) of 2D3 antibody on acute lymphocytic leukemia (ALL) patients was significantly lower than that (39/55) of acute myelogeneous leukemia (AML) (P <0.01). 2D3 was positive for all three cases of B/myeloid mixed lineage leukemia, but negative in 6 cases of chronic myelogeneous leukemia (CML), significantly lower than that in AML (39/55, P<0.01) patients. There was no difference between the positive rates of chronic lymphocytic leukemia (CLL, 3/5) and B lineage ALL (9/28, P = 0.2389). The antibody was not found to recognize the differentiation stages of either ALL and AML subtypes.</p><p><b>CONCLUSION</b>2D3 monoclonal antibody primarily recognizes AML cases. Further studies on cloning of gene encoding the antigen protein and its biological functions are warranted.</p>


Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Antibodies, Monoclonal , Allergy and Immunology , Antibodies, Neoplasm , Allergy and Immunology , Antigens, Neoplasm , Allergy and Immunology , Immunophenotyping , Leukemia, Lymphocytic, Chronic, B-Cell , Allergy and Immunology , Leukemia, Myeloid, Acute , Allergy and Immunology , Leukocyte Common Antigens , Allergy and Immunology , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Allergy and Immunology
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