Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 27
Filter
1.
International Eye Science ; (12): 379-384, 2023.
Article in Chinese | WPRIM | ID: wpr-964233

ABSTRACT

AIM: To study the role and mechanism of curcumol in neovascularization induced by vascular endothelial growth factor(VEGF).METHODS: Human umbilical vein endothelial cells were cultured in vitro and treated with 50ng/mL VEGF and curcumol at different concentrations. Cell proliferation was detected by CCK-8 and EdU assay, the migration ability of cells was analyzed by Transwell assay, the angiogenesis ability of endothelial cells was analyzed by tube formation assay, and the change of Akt/mTORC1 signal pathway was detected by Western blot.RESULTS: CCK-8 results showed that the OD450 value of cells in 400 and 800 μmol/L curcumol+VEGF group was significantly lower than that in VEGF group(all P<0.01). EdU results showed that the rate of cell proliferation in 400 μmol/L curcumol+VEGF group was significantly lower than that in VEGF group(P<0.001). Transwell assay and the formation assay results showed that the number of migratory cells in 400 μmol/L curcumol+VEGF group was decreased, and the number and length of tube branches were also reduced compared with VEGF group(all P<0.001). Western blot results showed that curcumol significantly inhibited the expression of p-Akt and p-S6, which were downstream targets of Akt/mTORC1 pathway in cells.CONCLUSION: Curcumol can inhibit VEGF-induced cell proliferation, migration and tube formation of vein endothelial cells, and has a strong inhibitory effect on angiogenesis, which can be further studied in the treatment of ocular fundus neovascularization.

2.
Journal of Experimental Hematology ; (6): 944-950, 2021.
Article in Chinese | WPRIM | ID: wpr-880173

ABSTRACT

OBJECTIVE@#To investigate the clinical characteristics and risk factors of cytomegalovirus (CMV) infection after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with severe aplastic anemia (SAA).@*METHODS@#Clinical data from 270 SAA patients with allo-HSCT were retrospectively analyzed, including 108 sib congruence patients and 162 substitute donors (68 unrelated donor congruence patients and 94 related haploid patients). Different pretreatment schemes were selected for different transplantation modes. The HLA-identical sibling and haploid grafts were all bone marrow and peripheral blood stem cells, and the grafts from unrelated donors were peripheral blood stem cells. After granulocyte implantation, blood CMV-DNA was regularly monitored. Flow cytometry was also used to determine the absolute number of CD3@*RESULTS@#CMV infection occurred in 229 of 270 patients with an incidence of 84.8%. Among them, 18 patients developed giant cell disease. Univariate analysis showed that alternative donors (unrelated total and haploid donors), mycophenolate mofetil and acute graft-versus-host disease were statistically significantly associated with CMV infection (P<0.05). Multivariate analysis showed that alternative donors were associated with CMV infection. The recovery of CD3@*CONCLUSION@#After allo-HSCT, substitute donors are more easily to develop CMV infection than full-sibling donors, and the reconstruction of immune function is delayed after transplantation.


Subject(s)
Humans , Anemia, Aplastic , Cytomegalovirus Infections , Graft vs Host Disease , Hematopoietic Stem Cell Transplantation , Retrospective Studies
3.
China Journal of Chinese Materia Medica ; (24): 4959-4968, 2021.
Article in Chinese | WPRIM | ID: wpr-921633

ABSTRACT

Azadirachtin, as a botanical insecticide, is a highly oxidized limonoid triterpenoid existing in the seeds of Azadirachta indica. However, due to the low content in the seeds, the production of azadirachtin by seed extraction has low yield. Chemical synthesis of azadirachtin is characterized by complex process and low yield. Synthetic biology provides an alternative for the supply of azadirach-tin. In this study, two oxidosqualene cyclases AiOSC1 and MaOSC1 respectively derived from A. indica and Melia azedarach were identified in yeast. A yeast strain producing tirucalla-7,24-dien-3β-ol was constructed by integration of AiOSC1, Arabidopsis thaliana-derived squalene synthase gene(AtAQS2), and Saccharomyces cerevisiae-derived truncated 3-hydroxy-3-methyl-glutaryl coenzyme A reductase gene(PgtHMGR) into the delta site of yeast. Then, the function of MaCYP71BQ5 was successfully verified in yeast after this gene was introduced into the constructed yeast strain. This study not only laid a foundation for the biosynthesis of tirucalla-7,24-dien-3β-ol, but also provided a chassis cell for the functional identification of cytochrome oxidases(CYP450 s) in azadirachtin biosynthesis pathway.


Subject(s)
Azadirachta , Limonins , Saccharomyces cerevisiae/genetics , Triterpenes
4.
China Journal of Chinese Materia Medica ; (24): 3819-3825, 2020.
Article in Chinese | WPRIM | ID: wpr-828380

ABSTRACT

In this study, citrate synthase gene(CIT2), and malate synthase gene(MLS1) were successfully knocked out in β-amyrin-producing yeast cells by using CRISPR/CAS9. The promoter of phosphoglucose isomerase gene(PGI1) was replaced by that of cytochrome c oxidase subunit Ⅶa(Cox9)to weaken its expression, aiming to channel more carbon flux into the NADPH-producing pathway. The fermentation results showed that CIT2 deletion had no effect on the β-amyrin production. Compared with the control strain, the production of β-amyrin was increased by 1.85 times after deleting MLS1, reaching into 3.3 mg·L~(-1). By replacing the promoter of PGI1, the β-amyrin yield was 3.75 times higher than that of the control strain, reaching up to 6.7 mg·L~(-1). This study successfully knocked out the CITT2 and MLS1 genes and weakened the PGI1 gene by using CRISPR/CAS9, which directly influenced the production of β-amyrin and provided some reference for the the metabolic engineering of triterpernoid producing strain.


Subject(s)
Ethanol , Fermentation , Metabolic Engineering , Saccharomyces cerevisiae , Genetics
5.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 63-67, 2019.
Article in Chinese | WPRIM | ID: wpr-802420

ABSTRACT

Objective: To observe the clinical efficacy modified Shenling Baizhu San in treating patients with spleen deficiency wetness type subacute eczema, and its effect on the inflammatory factors and peripheral blood T lymphocyte subsets, so as to provide a theoretical basis for clinical treatment. Method: Totally 176 cases of spleen deficiency wetness type subacute eczema treated at our hospital from Jan. 2016 to Dec. 2017 were selected and randomly divided into control group (88 cases) and observation group (88 cases). Control group was treated with levocetirizine hydrochloride tablets and compound econazole nitrate gel, and observation group was given modified Shenling Baizhu San in addition to therapy of control group for 4 weeks. The clinical efficacy, pruritus score, severity index(EASI)score, dermatology quality of life scales(DQOLS) score and recurrence rate were observed,and inflammatory factor and peripheral blood T lymphocyte subgroup level were detected. Result: After treatment, the pruritus score and EASI score in observation group were lower than those before treatment (PPPPβ, IL-2, IL-4, IL-5 and tumor necrosis factor(TNF) -α in observation group were lower than those before treatment (P+ and CD4+/CD8+ were higher than those of before treatment, and significantly higher than those of control group (PPConclusion: Modified Shenling Baizhu San could enhance the clinical efficacy by inhibiting the inflammatory reaction and regulating the immune function of patients.

6.
China Journal of Chinese Materia Medica ; (24): 1341-1349, 2019.
Article in Chinese | WPRIM | ID: wpr-774550

ABSTRACT

In this study, the synthetic pathway of β-amyrin was constructed in the pre-constructed Saccharomyces cerevisiae chassis strain Y0 by introducing β-amyrin synthase from Glycyrrhiza uralensis, resulting strain Y1-C20-6, which successfully produced β-amyrin up to 5.97 mg·L~(-1). Then, the mevalonate pyrophosphate decarboxylase gene(ERG19), mevalonate kinase gene(ERG12), 3-hydroxy-3-methylglutaryl-CoA synthase gene(ERG13), phosphomevalonate kinase gene(ERG8) and IPP isomerase gene(IDI1)were overexpressed to promoted the metabolic fluxto the direction of β-amyrin synthesis for further improving β-amyrin production, resulting the strain Y2-C2-4 which produced β-amyrin of 10.3 mg·L~(-1)under the shake flask fermentation condition. This is 100% higher than that of strain Y1-C20-6, illustrating the positive effect of the metabolic engineering strategy applied in this study. The titer of β-amyrin was further improved up to 157.4 mg·L~(-1) in the fed-batch fermentation, which was almost 26 fold of that produced by strain Y1-C20-6. This study not only laid the foundation for the biosynthesis of β-amyrin but also provided a favorable chassis strain for elucidation of cytochrome oxidases and glycosyltransferases of β-amyrin-based triterpenoids.


Subject(s)
Fermentation , Glycyrrhiza uralensis , Genetics , Industrial Microbiology , Intramolecular Transferases , Genetics , Metabolic Engineering , Oleanolic Acid , Saccharomyces cerevisiae , Metabolism
7.
Chinese Journal of Infection Control ; (4): 153-157, 2019.
Article in Chinese | WPRIM | ID: wpr-744323

ABSTRACT

Objective To understand the occurrence regularity and epidemic features of notifiable infectious diseases (NIDs) among children in a tertiary general hospital, provide scientific basis for the triage and referral of infectious diseases, as well as formulation of prevention and control measures of healthcare-associated infection (HAI) among children in a general hospital.Methods Descriptive epidemiological method was used to analyze the epidemiological data of reported NIDs in children in the hospital from 2013 to 2017.Results From 2013 to 2017, 1 170 children with infectious diseases were reported, the average annual reporting rate was 5.81‰, 670 cases (57.26%) were males and 500 (42.74%) were females.The population distribution was mainly students (n=503, 42.99%) and scatter lived children (n=433, 37.01%).The reported cases were mainly concentrated in the second quarter of each year, the top three diseases were chickenpox (n=423, 36.15%), hand-foot-mouth disease (n=332, 28.38%), and mumps (n=199, 17.01%).Conclusion Infectious disease in children is an important link in the prevention and control of infection in tertiary general hospitals, the report of infectious diseases of hand-foot-mouth disease, chickenpox and mumps, as well as implementation of prevention and control measures of HAI should be strengthened.

8.
China Journal of Chinese Materia Medica ; (24): 2421-2432, 2019.
Article in Chinese | WPRIM | ID: wpr-773243

ABSTRACT

With the development of various biotechnology,the research on molecular genetics of medicinal plants has gradually deepened. In this paper,the research system of molecular genetics of medicinal plants was proposed for the first time,which was elaborated from the aspects of genetic resources,genome,gene function and research methods. The application fields of medicinal plant mainly contain species identification,molecular breeding and biosynthesis. The research directions of molecular genetics of medicinal plants in genetic resources,model platform,synthetic biology and molecular breeding were put forward,which include 1 000 genome projects of medicinal plants,model species and mutant libraries,gene original libraries of heterologous synthetic systems,construction gene original library and specific chassis cells in heterologous synthesis system of active ingredient,breeding of new varieties of medicinal plants with high active ingredient and high resistance based on molecular markers andtransgenes.


Subject(s)
Biotechnology , Gene Library , Genetic Markers , Genome, Plant , Molecular Biology , Plant Breeding , Plants, Medicinal , Genetics , Research , Transgenes
9.
China Journal of Chinese Materia Medica ; (24): 86-91, 2018.
Article in Chinese | WPRIM | ID: wpr-776419

ABSTRACT

Residue of Mori Cortex was studied to optimize its enzymatic hydrolysis process, and explore its potential as a carbon source for biochemistry and biofuel production. The cellulose content of diluted acid pretreated (DAP) and non-pretreated from Mori Cortex were measured in this study, and the results showed that the cellulose content of DAP and non-pretreated from Mori Cortex were 52.5% and 47%, respectively. This higher cellulose content indicated that residue of Mori Cortex had the potential to act as a carbon source for biochemistry and biofuel production. Enzymatic hydrolysis of pretreated and non-pretreated from Mori Cortex was conducted under different enzyme loading amount. 40 FPU·(g DW)⁻¹ enzyme loading was determined as the optimal amount by comparing the yield of sugar and the rate of enzymolysis. Under this condition, the concentrations of glucose, xylose, arabinose sugar were 23.82, 4.84, 3.6 g·L⁻¹, and the corresponding enzymatic hydrolysis rate was 45.33% which was 2.3 times higher than that of non-pretreated from Morus alba residues. Fed-batch enzymatic hydrolysis was conducted finally to get higher sugar yield, and the final glucose concentration reached up to 38 g·L⁻¹ with the enzymatic hydrolysis rate of 36.19%. The results indicated that Mori Cortex residue had higher cellulose and hemicellulose contents, so it had the potential to become a carbon source to produce the bio-chemicals and biofuels. Through enzymatic hydrolysis, it can be converted into microbial available monosaccharides; and through fermentation, it can be converted into high value-added chemicals, biofuels, etc., to solve the problem of residue pollution, and achieve the sustainable development and greening of Chinese pharmaceutical production process.


Subject(s)
Carbohydrates , Cellulose , Chemistry , Enzymes , Metabolism , Fermentation , Hydrolysis , Morus , Chemistry
10.
Journal of Preventive Medicine ; (12): 148-152, 2018.
Article in Chinese | WPRIM | ID: wpr-792710

ABSTRACT

Objective To assess the dietary risk of organophosphates pesticides residues in some vegetables in Shaanxi Province. Methods The monitoring data of organophosphorus pesticides in vegetables from 2012 to 2016 were derived from Shaanxi Province Food Contamination Monitoring Network. The exposure of organophosphorus to vegetables by using exposure risk index (ERI) , dietary exposure risk index (RI) and dietary exposure risk assessment. Results Exposed risk index (ERI) of organophosphorus in some vegetables in Shaanxi Province ranged from 1.25E-06 to 1.87E-01, and among them, the highest ERI of clozaptid in garlic was 1.87E-01, followed by onion. In some vegetables, the highest exposure to organophosphorus was isocarbophos of fresh beans with an exposure of 4.27E-02 μg / (kg body weight · day) and the lowest dietary exposure was bulbs, and stems and fresh beans were 2.00E-04μg / (kg body weight·day) . The exposure of organophosphorus to all kinds of vegetables was less than their respective daily allowable intake (ADI) . The risk of dietary exposure to organic- phosphorus in vegetables was 187% and greater than 100% for the risk index (RI) of lettuce, and RI for all other vegetables was <100%. Conclusion The dietary exposure and risk index of organophosphates pesticides in some vegetables in Shaanxi Province was safe, and more attention should be paid to dimethoate and isazofos.

11.
China Journal of Chinese Materia Medica ; (24): 4124-4128, 2016.
Article in Chinese | WPRIM | ID: wpr-272723

ABSTRACT

Flavonoids are the valuable components in medicinal plants, which possess a variety of pharmacological activities, including anti-tumor, antioxidant and anti-inflammatory activities. There is an unambiguous understanding about flavonoids biosynthetic pathway, that is,2S-flavanones including naringenin and pinocembrin are the skeleton of other flavonoids and they can transform to other flavonoids through branched metabolic pathway. Elucidation of the flavonoids biosynthetic pathway lays a solid foundation for their synthetic biology. A few flavonoids have been produced in Escherichia coli or yeast with synthetic biological technologies, such as naringenin, pinocembrin and fisetin. Synthetic biology will provide a new way to get valuable flavonoids and promote the research and development of flavonoid drugs and health products, making flavonoids play more important roles in human diet and health.

12.
China Journal of Chinese Materia Medica ; (24): 4129-4137, 2016.
Article in Chinese | WPRIM | ID: wpr-272722

ABSTRACT

Catharanthus roseus can produce a variety of terpenoid indole alkaloids (TIA), most of which exhibit strong pharmacological activities. Hence, biosynthesis and regulation of TIA have received recent attention. 3α (S)-strictosidine is an important node in TIA biosynthesis, which is a condensation product of secologanin and tryptamine. The former is produced in iridoid pathway, and the latter is produced in indole pathway. Vindoline and catharanthine, which are produced respectively by 3α (S)-strictosidine via multi-step enzymatic reaction, can form α-3, 4-anhydrovinblastine by the condensation reaction. Then, vinblastine and vincristine are generated from α-3, 4-anhydrovinblastine. Many transcription factors are involved in the regulation of TIA synthesis, such as AP2/ERF and WRKY. Illumination of biosynthetic pathway has laid a foundation for the study of synthetic biology. Today, 3α (S)-strictosidine and vindoline have been synthesized in heterologous hosts Saccharomyces cerevisiae.Research about synthetic biology and the regulation mechanisms will provide a guidance for the production and development of TIA drugs in C. roseus.

13.
China Journal of Chinese Materia Medica ; (24): 4144-4149, 2016.
Article in Chinese | WPRIM | ID: wpr-272720

ABSTRACT

Taxol, a kind of terpenoid secondary metabolite produced by Taxus brevifolia, is an effective anticancer drug that manufacture relies mainly on the extraction form plants. In order to solve the resource shortage, a lot of work has been done to develop the alternative method. Recently, using synthetic biology to realize heterologous biosynthesis of the precursors of taxol has become a hotspot. Now, the basic framework of taxol biosynthetic pathways has been confirmed, and most enzyme genes involved in taxol biosynthesis have been cloned and identified. The two taxol precursors, taxa-4(5),11(12)-diene and taxa-4(20),11(12)-dien-5α-ol, have been synthesized in Escherichia coli and Saccharomyces cerevisiae. Here this paper reviewed the recent advances in the biosynthetic pathway of taxol and the latest developments of synthetic biology, which aims to provide a guidance for the heterologous biosynthesis of taxol.

14.
China Journal of Chinese Materia Medica ; (24): 4150-4157, 2016.
Article in Chinese | WPRIM | ID: wpr-272719

ABSTRACT

The functional ingredients in Chinese materia medica are the main active substance for traditional Chinese medicine and most of them are secondary metabolites derivatives. Until now,the main method to obtain those functional ingredients is through direct extraction from the Chinese materia medica. However, the income is very low because of the high extraction costs and the decreased medicinal plants. Synthetic biology technology, as a new and microbial approach, can be able to carry out large-scale production of functional ingredients and greatly ease the shortage of traditional Chinese medicine ingredients. This review mainly focused on the recent advances in synthetic biology for the functional ingredients production.

15.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 71-75, 2013.
Article in Chinese | WPRIM | ID: wpr-355587

ABSTRACT

<p><b>OBJECTIVE</b>Based on proteomics technology, Pi-yang deficiency syndrome (PYDS) correlated differential proteins were screened, thus providing powerful experiment reliance for exploring the essence of PYDS.</p><p><b>METHODS</b>Totally 36 SD rats of SPF grade were randomly divided into the normal control group (n = 16) and the PYDS group (n = 20). The PYDS model rats were induced by improper diet, overstrain, and administration of yang impairing bitter cold herbs. The total proteins of the ileum were separated and extracted from rats in the PYDS group and the normal control group. The differential protein dots were identified using Delyder 2D 6.5 image analysis software by two-dimensional gel electrophoresis (2-DE) technology. The finger print map of corresponding peptide qualities was obtained by applying MALDI TOF/TOF. The differential proteins were identified using Mascot search library.</p><p><b>RESULTS</b>Judged by statistics and fuzzy mathematics, Pi-yang deficiency rat model was successfully established. Eight proteins with differential expressions involving cell skeleton, energy metabolism, and signal transduction, and so on were obtained. Of them, there were 4 up-regulated proteins, i.e., desmin, cytokeratin8 (CK8), pyruvate kinase (PK), and ezrin. Four down-regulated proteins were glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cytokeratin19 (CK19), cytokeratin1 (CK1), and actin.</p><p><b>CONCLUSION</b>The pathogenesis of PYDS might be slowed energy metabolism rate, reduced energy production, changed structure of ileal villin, and weakened absorbing and digestive functions.</p>


Subject(s)
Animals , Female , Male , Rats , Ileum , Metabolism , Medicine, Chinese Traditional , Proteome , Metabolism , Proteomics , Yang Deficiency , Diagnosis , Metabolism
16.
Chinese Pharmaceutical Journal ; (24): 1733-1736, 2013.
Article in Chinese | WPRIM | ID: wpr-860192

ABSTRACT

OBJECTIVE: To explore the anti-tumor efficacy of liposomal vinorelbine tartrate injection(NVB-lipo). METHODS: The anti-neoplastic effect of NVB-lipo was evaluated by using mice bearing H22 tumor and nude mice bearing NCI-H460 human xenograft tumor model. RESULTS: After a single intravenous injection at doses of 20, 10, 5 and 2.5 mg · kg-1, the inhibition ratio of H22 tumor were 82.1%, 75.8%, 63.2% and 35.4% respectively, and that of NVB-free(20 and 10 mg · kg-1) were 45.8% and 37.1%. After three times iv injection interval 3-day, NVB-lipo 8 mg · kg-1 could markedly inhibited the growth of NCI-H460 tumor, relative to control and NVB-free (P < 0.05). The anti-tumor effect of NVB-lipo in H22 and NCI-H460 tumor model was much stronger than NVB-free, and was positively correlated with its dose levels. CONCLUSION: At the same dose level, the antineoplastic effects of NVB-lipo is stronger than NVB-free.

17.
Acta Pharmaceutica Sinica ; (12): 1565-1569, 2010.
Article in Chinese | WPRIM | ID: wpr-250594

ABSTRACT

This study is to compare the pharmacodynamics, pharmacokinetics and tissue distribution of liposomal mitoxantrone (Mit-lipo) and free mitoxantrone (Mit-free). The antineoplastic effect of Mit-lipo was evaluated on PC-3 human xenograft tumor model after repeated intravenous injection at dose levels of 1, 2 and 4 mg x kg(-1). The pharmacokinetic study of Mit-lipo and Mit-free was performed on dogs following a single intravenous injection. The tissue distribution of Mit-lipo and Mit-free was observed on S-180 bearing mice after a single intravenous injection. (1) Pharmacodynamics: Mit-lipo dose-dependently inhibited PC-3 tumor growth at a dose ranging from 1 to 4 mg x kg(-1). The antitumor effect studies showed that Mit-lipo significantly improved the therapeutic effect in comparison with free drug. (2) Pharmacokinetics: in comparison with Mit-free, the AUC and t(1/2) values of Mit-lipo at the same dose level were higher than those of Mit-free in Beagle dogs. The results showed that Mit-lipo had long circulation characteristics. (3) Tissue distribution in S-180 bearing mice: compared to Mit-free, Mit-lipo preferentially accumulated into tumor zones instead of normal tissues. Tumor AUC in Mit-lipo treated animals was 8.7 fold higher than that in mice treated with the same dose of Mit-free. The Cmax values of Mit-lipo in heart, kidney, lung, spleen and intestinal tissue in Mit-lipo were 30.2%, 161.6%, 20.2%, 27.9% and 78.3% lower than those of Mit-free, respectively. The pharmacokinetics and tissue distribution of Mit-lipo changed obviously, thus increasing therapeutic effect and improving drug therapeutic index.


Subject(s)
Animals , Dogs , Female , Humans , Male , Mice , Antineoplastic Agents , Pharmacokinetics , Pharmacology , Area Under Curve , Cell Line, Tumor , Cell Proliferation , Dose-Response Relationship, Drug , Drug Carriers , Injections, Intravenous , Liposomes , Chemistry , Mice, Inbred BALB C , Mice, Nude , Mitoxantrone , Pharmacokinetics , Pharmacology , Neoplasm Transplantation , Prostatic Neoplasms , Pathology , Sarcoma 180 , Pathology , Tissue Distribution
18.
Journal of Experimental Hematology ; (6): 450-453, 2010.
Article in Chinese | WPRIM | ID: wpr-243337

ABSTRACT

The aim of this study was to detect DNA damage during expansion ex vivo of umbilical cord blood (UCB) hematopoietic cells and explore the optimal harvest time for culture of CB hematopoietic cells. Mononuclear cells (MNCs) separated from UCB were cultured in a serum-free system supplemented with cytokines and colony forming units were assessed by semisolid culture at the same time. On day 0, 7, 14 and 21 cells were collected for single cell gel electrophoresis (SCGE) analysis and CFUs were also assayed by SCGE, CD34+ cells and CD133+ cells were quantitated by fluorescence-activated cell sorting (FACS). The results showed that the percentage of CD34+ and CD133+ cells was found to be highest after short-term culture (<14 days) and the cord blood DNA damage rate was observed to be less than 5.0% at earlier time points, but at day 21 the DNA damage rate was 28.2%, which was higher than that at day 0 (p=0.000), the tail length of the DNA comet was longer than that at day 0 (p=0.000). The tail lengths of DNA damage on other time points were not significantly different from that at day 0. It is concluded that the DNA damage rate is less than 5.0% after short-term (<14 days) culture of UCB cells ex vivo by using this method. After 14 days DNA damage rate increases significantly. The optimal harvest time of cord blood cells after culture ex vivo would be within 14 days.


Subject(s)
Humans , Cell Division , Cells, Cultured , Colony-Forming Units Assay , DNA Damage , Fetal Blood , Cell Biology , Hematopoietic Stem Cells , Cell Biology
19.
Journal of Experimental Hematology ; (6): 431-436, 2009.
Article in Chinese | WPRIM | ID: wpr-302117

ABSTRACT

This study was purposed to investigate the expression and clonal proliferation of receptor (TCR) Vbeta subfamilies of the T-cells in acute leukemic patients at different disease status (onset, complete remission or relapse) and to analyze the influence of the leukemic cell load on anti-leukemic effect of peripheral T-lymphocytes of the patients. Gene sequences of peripheral TCR Vbeta 24 families from 11 leukemic patients and 3 normal donors were expanded by RT-PCR. Genescan technique was applied to evaluate clonal expression of the TCRVbeta subfamilies, clonal characteristics of the CDR3 from peripheral blood of AML patients at different disease status. The application, clonal proliferation, cellular complexity of T-cells, and the variation of immunotypes of T-cells were compared. The results indicated that the lower and partial distribution of TCR Vbeta subfamily was found in all 11 patients when firstly diagnosed; the expression of TCR Vbeta subfamilies after induction in vitro increased; obvious elevation of TCR Vbeta subfamilies was observed in patients at complete remission although expression level was still lower than normal, whereas the significant descent of TCR Vbeta subfamilies was detected in 4 relapsed patients. Only 1 - 2 clonal proliferation of TCR Vbeta subfamilies existed in 9 out of 11 patients at initial diagnosis which increased at remission. The status of clonal proliferation of Vbeta subfamily T-cells continued regardless of any different disease status in most patients. There was an obvious decrease of CDR3 complexity at initial diagnosis or relapse, while CDR3 complexity would be partially improved at remission. It is concluded that the restrict distribution and expression of TCR Vbeta subfamilies were found in AML patients. Clonal proliferation of T-cells Vbeta subfamily continuously exists regardless of any different disease status in most patients. Some Vbeta subfamilies sustain clonal proliferation at different disease status. Some clonal proliferations of Vbeta subfamilies are associated with the effects of leukemic cells, CDR3 complexity obviously decreases under disease status which can be partially improved at remission.


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Clone Cells , Complementarity Determining Regions , Genetics , Leukemia, Myeloid, Acute , Genetics , Receptors, Antigen, T-Cell , Genetics , Reverse Transcriptase Polymerase Chain Reaction
20.
Journal of Experimental Hematology ; (6): 1511-1515, 2009.
Article in Chinese | WPRIM | ID: wpr-328610

ABSTRACT

This study was aimed to investigate the effect of 1,4-benzoquinone (1,4-BQ) on growth of myeloid progenitor cells with IFN-gamma different genotypes and to compare its differences. Polymerase chain reaction (PCR) was used to amplify the polymorphism gene segment of IFN-gamma +874 A/T in 36 cord blood (CB) specimens. The specimens were divided into three groups (AA, AT and TT group). MNCs were planted on complete methylcellulose medium containing different concentrations of 1,4-BQ. The colony-forming units (CFU) were assayed, the differences of colony growth in specimens with different genotypes (AA, AT and TT) under 1,4-BQ exposure were analyzed. The results showed that frequencies of AA, AT and TT genotypes were 5.56%, 88.89% and 5.56% in the 36 CB samples respectively. Comparing colony numbers of IFN-gamma +874 AA, AT and TT genotype indicated that there was significant difference (p(AA) = 0.033, p(AT) = 0.009, p(TT) = 0.001, < 0.05). Significant cytotoxicity was observed after exposure to concentrations of 1,4-BQ > or = 5 micromol/L. Cytotoxic response of 1,4-BQ was dose-dependent. Under the same concentration of 1,4-BQ, there were no significant differences in capacity of cell colony growth between 3 groups (AA, AT and TT). Colony numbers of specimen No 3 in AT group and specimen No 2 in TT group were less than those of other specimens significantly. It is concluded that the hematopoietic myeloid progenitor cells cultured in the presence of 1,4-BQ show a dose-dependent cytotoxic response, but there are no significant differences in colony growth of IFN-gamma different genotypes (AA, AT and TT) under the same concentration of 1,4-BQ.


Subject(s)
Humans , Benzoquinones , Pharmacology , Bone Marrow Cells , Fetal Blood , Cell Biology , Genotype , Hematopoietic Stem Cells , Interferon-gamma , Genetics , Stem Cells
SELECTION OF CITATIONS
SEARCH DETAIL