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1.
Article in English | WPRIM | ID: wpr-939907

ABSTRACT

Bacterial surface glycans perform a diverse and important set of biological roles, and have been widely used in the treatment of bacterial infectious diseases. The majority of bacterial surface glycans are decorated with diverse rare functional groups, including amido, acetamidino, carboxamido and pyruvate groups. These functional groups are thought to be important constituents for the biological activities of glycans. Chemical synthesis of glycans bearing these functional groups or their variants is essential for the investigation of structure-activity relationships by a medicinal chemistry approach. To date, a broad choice of synthetic methods is available for targeting the different rare functional groups in bacterial surface glycans. This article reviews the structures of naturally occurring rare functional groups in bacterial surface glycans, and the chemical methods used for installation of these groups.


Subject(s)
Bacterial Infections , Humans , Polysaccharides/chemistry , Structure-Activity Relationship
2.
Article in English | WPRIM | ID: wpr-929269

ABSTRACT

Most bacterial cell surface glycans are structurally unique, and have been considered as ideal target molecules for the developments of detection and diagnosis techniques, as well as vaccines. Chemical synthesis has been a promising approach to prepare well-defined oligosaccharides, facilitating the structure-activity relationship exploration and biomedical applications of bacterial glycans. L-Galactosaminuronic acid is a rare sugar that has been only found in cell surface glycans of gram-negative bacteria. Here, an orthogonally protected L-galactosaminuronic acid building block was designed and chemically synthesized. A synthetic strategy based on glycal addition and TEMPO/BAIB-mediated C6 oxidation served well for the transformation of commercial L-galactose to the corresponding L-galactosaminuronic acid. Notably, the C6 oxidation of the allyl glycoside was more efficient than that of the selenoglycoside. In addition, a balance between the formation of allyl glycoside and the recovery of selenoglycoside was essential to improve efficiency of the NIS/TfOH-catalyzed allylation. This synthetically useful L-galactosaminuronic acid building block will provide a basis for the syntheses of complex bacterial glycans.


Subject(s)
Carbohydrates , Glycosides , Oligosaccharides , Oxidation-Reduction , Polysaccharides/chemistry
3.
Article in Chinese | WPRIM | ID: wpr-905961

ABSTRACT

Objective:To explore the acute toxicities and hepatotoxicities of aqueous extracts of Taxilli Herba from <italic>Morus alba</italic>, <italic>Toxicodendron</italic> <italic>trichocarpum</italic>, <italic>Camellia oleifera</italic>, <italic>Salix babylonica</italic>, <italic>Melia azedarach</italic>, and <italic>Nerium indicum</italic> against zebrafish model and the effect of different hosts on the toxicity of Taxilli Herba, hoping to provide a theoretical basis for the safe use of Taxilli Herba. Method:The normally developed AB zebrafish at 3-day post fertilization was selected for acute toxicity study. According to the results of preliminary toxicity experiments, the zebrafishes were treated with aqueous extracts of Taxilli Herba from different hosts at six doses, and their mortality was calculated 72 h later. GraphPad Prism 6.0 was used for plotting the dose-toxicity curve, followed by the calculation of their median lethal concentration (LC<sub>50</sub>) and 10% lethal concentration (LC<sub>10</sub>). The gz15Tg/+(AB) liver fluorescent protein transgenic zebrafish with normal development at 4-day post fertilization was applied for the hepatotoxicity study. The zebrafishes were divided into the low-, medium-, and high-dose groups of aqueous extracts of Taxilli Herba from six hosts, the positive control (acetaminophen) group, and the blank (embryo amniotic fluid) group, and then treated with the corresponding drugs. Seventy-two hours later, the liver morphology and fluorescent area changes in zebrafish were observed. And the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected. Result:The results of acute toxicity test demonstrated that the LC<sub>50</sub> values of water extracts of Taxilli Herba from <italic>M. alba</italic>, <italic>T.</italic> <italic>trichocarpum</italic>, <italic>C. oleifera</italic>, <italic>S. babylonica</italic>, <italic>M. azedarach</italic>, and <italic>N. indicum</italic> were 1.24, 0.94, 0.51, 0.38, 0.11, 0.09 g·L<sup>-1</sup>, respectively, and the LC<sub>10</sub> values were 0.70, 0.60, 0.35, 0.28, 0.08, 0.07 g·L<sup>-1</sup>, respectively. As revealed by hepatotoxicity test, compared with the blank group, the positive control group exhibited liver morphological changes, decreased fluorescent area (<italic>P</italic><0.01), and elevated ALT and AST activities (<italic>P</italic>< 0.01), suggesting that acetaminophen was hepatotoxic to zebrafish. However, there was no change in the liver morphology or fluorescent area of zebrafish in the low-, medium-, and high-dose groups of water extracts of Taxilli Herba from <italic>M. alba</italic>, and the ALT and AST activities were decreased. By contrast, the liver morphology and fluorescent areas in the medium- and high-dose groups of water extracts of Taxilli Herba from <italic>T.</italic> <italic>trichocarpum</italic>, <italic>C. oleifera</italic>, <italic>S. babylonica</italic>, <italic>M. azedarach</italic>, and <italic>N. indicum</italic> changed to varying degrees (<italic>P</italic><0.05, <italic>P</italic><0.01). Besides, the activities of both ALT and AST were also enhanced. These indicated that Taxilli Herba from <italic>M. alba</italic> had no hepatotoxicity to zebrafish, while that from <italic>T.</italic> <italic>trichocarpum</italic>, <italic>C. oleifera</italic>, <italic>S. babylonica</italic>, <italic>M. azedarach</italic>, and <italic>N. indicum</italic> showed varying degrees of hepatotoxicity to zebrafish. Conclusion:The toxicity of Taxilli Herba is host-dependent. Taxilli Herba from <italic>M. alba</italic> has no hepatotoxicity, but that from the other five hosts shows varying degrees of hepatotoxicity. Standardizing the host source may be an important measure to realize the medication safety of Taxilli Herba.

4.
Article in Chinese | WPRIM | ID: wpr-906440

ABSTRACT

Objective:To investigate the taxonomic structure and diversity of endophytic fungi from <italic>Datura metel </italic>and screen the strains with anti-dermatophyte activities, so as to provide resources for the development of new lead compounds against dermatomycosis. Method:Endophytic fungi were isolated from the roots, stems and leaves of <italic>D. metel</italic> after tissue block incubation and then identified by morphological analysis and rDNA-internal transcribed spacer(ITS) sequencing. Their anti-dermatophyte activities were detected by agar diffusion assay. Result:A total of 292 strains of endophytic fungi were isolated from <italic>D. metel</italic>, belonging to 34 genera, with<italic> Fusarium</italic> (72.97%) in roots, <italic>Fusarium </italic>(37.25%) and <italic>Plectosphaerella </italic>(28.43%) in stems, and <italic>Colletotrichum </italic>(39.66%) in leaves as the dominant species. The isolation rate (89.23%), colonization rate (84.62%), and diversity index (1.82) of endophytic fungi in leaves were significantly higher than those in roots (70.48%, 70.48% and 1.23) and stems (69.39%,68.03% and 1.64). The determination of anti-dermatophyte activities of 35 endophytic fungal fermented filtrates showed that the strains exhibiting inhibitory activities against <italic>Microsporum canis</italic>, <italic>Trichosporon mucoides</italic>, <italic>Trichophyton rubrum</italic> and <italic>Candida albicans </italic>accounted for 97.14%, 71.43%, 45.71%, and 25.71%, respectively. Among them, six strains (17.14%), namely <italic>Fusarium </italic>sp. R1, <italic>Penicillium </italic>sp. R5, <italic>Aspergillus </italic>sp. R7, <italic>Metarhizium </italic>sp.<italic> </italic>S18, <italic>Diaporthe </italic>sp. S19, and <italic>Glomerella </italic>sp.<italic> </italic>L57, all inhibited the four types of cutaneous fungal pathogens. Conclusion:The endophytic fungi in <italic>D. metel</italic> are diverse, and the proportion of endophytic fungi possessing anti-dermatophyte activities is high, allowing them to serve as potential resources for the development of new anti-dermatophyte agents.

5.
Journal of Experimental Hematology ; (6): 1610-1616, 2021.
Article in Chinese | WPRIM | ID: wpr-922304

ABSTRACT

OBJECTIVE@#To evaluate the incidence and clinical characteristics of metabolic syndrome (MS) within one year after hematopoietic stem cell transplantation (HSCT) in order to screen the risk factors for HSCT-MS, provide early intervention and improve the long-term quality of survival of patients.@*METHODS@#The clinical follow-up data of 64 HSCT patients (survival time > 1 year) who received HSCT in our center from January 2007 to August 2018 were collected. Among them, 50 cases were allogeneic hematopoietic stem cell transplantation (allo-HSCT) and 14 cases were autologous hematopoietic stem cell transplantation (auto-HSCT). The changes of MS-related indexes and clinical characteristics before and 1, 3, 6 and 12 months after HSCT were analyzed retrospectively.@*RESULTS@#In allo-HSCT group, 14 cases were diagnosed as MS before operation, including high-density lipoprotein cholesterol (hypo-HDL-C)> hyper triglycerides(hyper-TG)> hyper fasting glucose(hyper-FBG)> abdominal obesity (AO) > hypertension. The preoperative diagnosis of MS in the auto-HSCT group was 5 cases, in the order of hyper-FBG> hyper-TG> AO> hypo-HDL-C> hypertension. Incidence of MS at 1, 3, 6 and 12 months after transplantation: 19, 26, 24 and 20 cases in the allo-HSCT group, respectively; auto-HSCT group were 7, 7, 6 and 6 cases, respectively. Hyper-TG and hypo-HDL-C were prominent in both groups.@*CONCLUSION@#The incidence of HSCT-MS is significantly higher within 1 year after HSCT. Regardless of allo-HSCT and auto-HSCT, the prevention and control of HSCT-MS is emphasized as an important guarantee to improve the long-term survival quality of HSCT patients.


Subject(s)
Hematopoietic Stem Cell Transplantation , Hematopoietic Stem Cells , Humans , Metabolic Syndrome , Retrospective Studies , Transplantation, Homologous
6.
Article in Chinese | WPRIM | ID: wpr-743045

ABSTRACT

Objective To investigate the effect of extracts from rabbit skins inflamed by Viccinia virus vaccine (analgecine) on the proliferation of human cancer cells and on the cytokine secretion in mouse spleen lymphocytes in vitro. Methods Five human tumor cell lines, HepG2, LM3, H460, A549 and HeLa were used and the effect of analgecine (1.63, 0.815, 0.326, 0.163 and0.0815 U/ml) on the cell proliferation was evaluated by the CCK-8 assay. The mouse spleen was isolated aseptically, and the spleen lymphocyte suspension was prepared and cultured with PRMI-1640 medium containing 10% fetal bovine serum (FBS). For detection of the cytokine IL-2, IFN-γ, IL-4 and IL-12 level, the stimulant concanavalin A (ConA) or lipoplysaccharide (LPS) was added into the lymphocyte suspension, and the lymphocytes were cultured under the presence of analgecine at the final concentration of 0.815, 0.163 and 0.0815 U/ml for 24 hours. Then, the level of the cytokines in the supernatant was detected by the ELISA kit. On the other hand, the effect of supernatant of the spleen lymphocyte cultures under the presence of analgecine at 0.815 U/ml on the proliferation HepG2 cells was also evaluated by the CCK-8 assay. The CCK-8 assay was performed after cultivation of the HepG2 cells in the whole supernatant or in its dilution with fresh medium for 24 hours. Results Analgecine showed a dose-dependent inhibitory effect on the five tested cancer cell lines, with the inhibition rate of 58.95%, 55.08%, 57.28%, 45.80% and 48.18% at the 1.63 U/ml on the HepG2, LM3, H460, A549 and HeLa cells, respectively. Compared with the control group, the secretion of IL-2, IFN-γ and IL-4 was significantly increased in the 0.163 and 0.815 U/ml analgecine groups (P<0.01), while the secretion level of IL-12 was increased in the 0.0815, 0.163 and 0.815 U/ml analgecine groups (P<0.01). The supernatant of the mouse spleen lymphocyte cultures under the presence of0.163 U/ml analgecine could inhibit the HepG2 cell proliferation in a dose-dependent manner, and the inhibitory effect of the whole supernatant was significantly stronger than the effect of the same concentration analgecine 0.163 U/ml (P<0.01). Conclusion Analgecine could inhibit the cell proliferation of the tested five human cancer cell lines, increased the secretion of IL-2, IFN-γ, IL-4 and IL-12 cytokines in mouse spleen lymphocytes, all in vitro, and its effect on the cytokine secretion may be related to the inhibitory effect on the human cancer cell proliferation.

7.
Military Medical Sciences ; (12): 60-63, 2018.
Article in Chinese | WPRIM | ID: wpr-694317

ABSTRACT

Objective To investigate the effects of 72 h sleep deprivation(SD)on circadian clock gene expression in the rat liver.Methods Twelve rats were randomly divided into control group and SD group.An SD instrument was used to deprive the rats′sleep for 72 h.Then the abdominal cavity was exposed to obtain liver,and the expression of clock genes was detected by RT-PCR and Western blotting analysis, respectively.Results Compared with the control group, the mRNA levels of clock,npas2 and rev-erbαstrikingly decreased in the livers of the SD group rats.However,per1,per2 and rorαmRNA levels obviously increased.bmal1 and cry1 mRNA expression hardly changed in the control and SD groups. Meanwhile,the protein levels of liver BMAL1,CLOCK,NPAS2,CRY1 and REV-ERBαwere significantly down-regulated and PER1,PER2 and RORαprotein levels were up-regulated in SD group compared with control group.Conclusion 72 h SD can result in abnormal expressions of several circadian clock genes in the rat liver at both transcriptional and translational levels.

8.
China Medical Equipment ; (12): 130-133, 2018.
Article in Chinese | WPRIM | ID: wpr-706546

ABSTRACT

Objective: To design a hospital management information system based on attitude and perceived behavior control, and to explore the influence of the application of hospital management information system on the work performance of medical personnel. Methods: Through constructed hospital management information system and applied the work performance model of medical personnel to design questionnaire from three points of view included attitude, habits, subjective norm and perceived behavior control. And then the internal consistency and validity of the scale of measurement indexes of the questionnaire were tested, and the methods of statistical modeling and parameter estimation were adopted to carry out path analysis. Results: The results of primary estimation for the change situation of main medical indexes of application information system in earlier, middle and later stage indicated that the reliability of variates that were selected in this research was higher, and the consistency of interior of various variates was higher. At the three stages, earlier, middle and later stage, that information system was implemented, the person-time of leaving hospital, the number of inpatient surgery and the outpatient amount of expert displayed rising trend. And the drug proportion, average stay time decreased 9.45%, 3.36% and 16.16% and 4.27% respectively. Conclusion: Whether the hospital information system can be applied, the personal subjective factors of medical personnel play an important role.

9.
Military Medical Sciences ; (12): 894-897, 2017.
Article in Chinese | WPRIM | ID: wpr-694276

ABSTRACT

Objective To investigate the effects of 72 h sleep deprivation (SD) on circadian clock gene expression in the rat spleen.Methods The rats were randomly divided into control group and SD group.An SD instrument was used to deprive the rats of sleep for 72 h.Then the lymphocytes from the spleen were obtained by Ficoll seperation medium before the expression of clock genes was detected by RT-PCR and Western blotting analysis respectively.Results Compared with the control group,the mRNA levels of bmal1,clock,per2 and rev-erbα strikingly decreased in the spleens of the SD group rats.However,npas2,per1,rorα and cry1 mRNA expression hardly changed in the control and SD group.Meanwhile,the protein levels of spleen BMAL1,NPAS2,CRY1 and RORα were significantly down-regulated and PER1 protein levels were up-regulated in SD group compared with control group.However REV-ERBα protein expression remained unchanged in the control and SD group.Conclusion 72 h SD can result in abnormal expressions of several circadian clock genes in lymphocytes of the spleen at both transcription and translation levels.

10.
Article in Chinese | WPRIM | ID: wpr-693347

ABSTRACT

Objective To investigate the antiplatelet effects of candidate drug W1 when combined with omeprazole,respec?tively.Methods The experimental rats were randomly divided into 5 groups:control group,clopidogrel(10 mg/kg)group,clopido?grel(10 mg/kg)+omeprazole(80 mg/kg)group,W1(3 mg/kg)group,and W1(3 mg/kg)+omeprazole(80 mg/kg)group.Four hours after oral administration of the drugs,the rats were measured for their platelet aggregation rate;Western blot was used to deter?mine the protein expressions of P2Y12 receptor,P-Akt and P-Erk.Results For the platelet aggregation rate,compared with the con?trol group,the 4 groups decreased significantly(P<0.01);the platelet aggregation rate in the clopidogrel + omeprazole group in?creased significantly than that in the clopidogrel group(42% and 20.4%,respectively,P<0.01);the platelet aggregoction rate (30.9%)in W1+omeprazole group was significantly higher(P<0.01)than that in the W1 group(20.5%),which was lower than that in the clopidogrel+omeprazole group(P<0.01).For the protein expression detected by the western blotting,there were no signif?icant changes in the expression of P2Y12 receptor on the platelet surface,in the clopidogrel or W1 group in comparison with the clopi?dogrel+omeprazole or W1+omeprazole group,respectively,while the phosphorylation level of Akt and Erk was up-regulated in the clopidogrel+omeprazole or W1+omeprazole group compared with the clopidogrel or W1 group,and the up-ragulatory effect of omepra?zole was weaker in the W1+omeprazole group than that in the clopidogrel+omeprazole group. Conclusion Combined use of omepra?zole could inhibit the antiplatelet activities of clopidogrel or W1,with the inhibitory effect weaker in W1 group than in clopidogrel group,suggesting that the risk for the combination of omeprazole with W1 is likely less than that for the combination with clopidogrel.

11.
Article in English | WPRIM | ID: wpr-311355

ABSTRACT

<p><b>OBJECTIVE</b>Consuming phthalates may be due to the presence of food contact materials, such as plastic containers. In this study, we investigated the association between plastic container use and phthalate exposure in 2,140 Shanghai adults.</p><p><b>METHODS</b>Participants completed a questionnaire on the frequency of using plastic containers in different scenarios in the previous year (e.g., daily, weekly) and on the consumption of plastic-packaged foods in the previous three days (yes or no). Urinary phthalate metabolites were used to assess the association between phthalate exposure and the use of plastic containers.</p><p><b>RESULTS</b>The metabolites of di-(2-ethylhexyl) phthalate (DEHP) were the most frequently detected in urine. The results revealed that phthalate exposure was associated with consumption of plastic-packaged breakfast or processed food items in the previous three days. The consumption of these two food items had strong synergistic effects on increasing urinary concentrations of most phthalate metabolites.</p><p><b>CONCLUSION</b>Our results of plastic-packaged breakfast and processed food may be explained by the use of flexible plastic containers, indicating the importance of risk assessment for the application of flexible plastic containers.</p>


Subject(s)
Adult , China , Cities , Data Collection , Humans , Phthalic Acids , Metabolism , Urine , Plastics , Chemistry
12.
Article in Chinese | WPRIM | ID: wpr-230969

ABSTRACT

Sucrose non-fermenting 1-related protein kinase 2(SnRK2) plays a key role in abiotic stress signaling in plants. In this study, we cloned a SmSnRK2.4 gene belonging to subclass I of SnRK2 from Salvia miltiorrhiza by screening its transcriptome database. The SmSnRK2.4 gene contains 8 introns and 9 exons, with a 1 068 bp open reading frame encoding a polypeptide of 355 amino acids, the predicted molecular mass of which is 40.63 kDa. Prokaryotic expression of SmSnRK2.4 protein using pMAL-c2X as the expression vector displayed that the recombinant protein of SmSnRK2.4 gene in E. coli was consistent with the predicted size. A 3 000 bp promoter sequence of SmSnRK2.4 contained some stress-responsive elements and hormone-responsive elements. Quantitative real-time PCR analysis revealed that the expression of SmSnRK2.4 in root was much higher than that in stem and leaf, SmSnRK2.4 was strongly induced by PEG stress, weakly induced by ABA stress. This research provided a basis for further study of the SmSnRK2.4 gene playing the role in accumulate mechanism of secondary metabolites in S. miltiorrhiza under drought.

13.
Electron. j. biotechnol ; 15(4): 6-6, July 2012. ilus, tab
Article in English | LILACS | ID: lil-646956

ABSTRACT

Response surface methodology was used to optimize the fermentation conditions for the production of pristinamycin by immobilization of Streptomyces pristinaespiralis F213 in shaking flask cultivation. Seed medium volume, fermentation medium volume and shaking speed of seed culture were found to have significant effects on pristinamycin production by the Plackett-Burman design. The steepest ascent method was adopted to approach the vicinity of optimum space, followed by central composite design for further optimization. A quadratic model was built to fit the pristinamycin production. The optimum conditions were found to be seed medium volume of 29.5 ml, fermentation medium volume of 28.8 ml, and shaking speed of seed culture at 204 rpm. At the optimum conditions, a production of 213 mg/l was obtained, which was in agreement with the maximum predicted pristinamycin yield of 209 mg/l. This is the first report on pristinamycins production by immobilized S. pristinaespiralis using response surface methodology.


Subject(s)
Fermentation , Pristinamycin/biosynthesis , Streptomyces/metabolism , Culture Techniques
14.
Article in Chinese | WPRIM | ID: wpr-347571

ABSTRACT

<p><b>OBJECTIVE</b>To study serum insulin-like growth factor 1 (IGF-1) levels and their association with growth and development in infants aged 1-24 mouths.</p><p><b>METHODS</b>A total of 525 healthy infants (125 preterm, 400 term) were enrolled. Serum IGF-1 levels were measured using ELISA 1.5, 4, 6, 8, 12, 18 and 24 months after birth. The body weight and body length were simultaneously measured.</p><p><b>RESULTS</b>Serum IGF-1 levels were the lowest in preterm infants 1.5 months after birth (86+/-60 ng/mL). Thereafter, serum IGF-1 levels increased, and were significantly higher than those in term infants between 4 and 12 months after birth. Serum IGF-1 levels in term infants were the highest (116+/-52 ng/mL) 1.5 months after birth during their life of 12 months old. Thereafter, serum IGF-1 levels decreased and reached to a nadir (69+/-58 ng/mL) 8 months after birth. IGF-I levels were positively correlated with the weight and the height (SDS) in both preterm and term infants.</p><p><b>CONCLUSIONS</b>Serum IGF-1 levels are closely associated with growth and development in infants.</p>


Subject(s)
Body Height , Body Weight , Child Development , Female , Humans , Infant , Infant, Newborn , Infant, Premature , Insulin-Like Growth Factor I , Male
15.
Article in Chinese | WPRIM | ID: wpr-671357

ABSTRACT

Objective To investigate the relation between the cellular immune function and unexplained recurrent spontaneous abortion(URSA) and the mechanism of active immunotherapy on URSA patients.Methods The flow cytometry(FCM) was used to detect CD3+ ,CD4+ ,CD8+ T lymphocyte and CD16+CD56+ natural killer(NK) cell subsets and the ratio of CD4+/CD8+ of peripheral blood(PBL).112 cases with URSA(76 cases treated with active immunotherapy) and 30 cases of normal fertiled(NF) women were studied.The percentages of T lymphocyte and NK cell subsets before and after therapy were compared among 76 cases wtih URSA treated by active immunotherapy.The rate of next successful pregnancy of URSA patients treated with and without active immunotherapy was compared.Results The percentages of CD3+ and CD16+CD56+ cell subsets as well as the ratio of CD4+/CD8+ of the URSA patients were significantly higher than those of NF cases(P <0.05).After active immunotherapy,the percentages of CD3+ and CD56+CD16+ cell subsets as well as the ratio of CD4+/CD8+ of URSA cases were significantly decreased(P <0.05 ).The rate of next successful pregnancy of URSA cases with and without active immunotherapy were 88.2% and 31.2% respectively,the difference was significant(P< 0.05).Conclusion The changes in the percentages of T lympyocyte and NK cell subsets have something to do with URSA.Active immunotherapy can effectively regulate the cellular immune function and increase the rate of next successful pregnancy of URSA patients.

16.
Journal of Experimental Hematology ; (6): 1408-1412, 2008.
Article in Chinese | WPRIM | ID: wpr-234223

ABSTRACT

This study was aimed to investigate the effects of xenogeneic antigen neu-Fc in combination with the recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF) and Bacillus Calmette-Guerin (BCG) on the regulation of Th1 and Th2 immune response in vitro. The rat neu L2-S2 domain was engineered as a chimeric protein with human IgG Fc. The eukaryotic expression vector was constructed. The recombinant protein was stably expressed in CHO cells and purified by rProtein A Sepharose Fast Flow column. The recombinant protein was identified by SDS-PAGE and Western blot. Peripheral blood mononuclear cells (PBMNCs) were obtained by means of standard Ficoll separation from the blood of healthy donors. Neu-Fc-induced PBMNC proliferation was tested by MTT. The production of IL-12 and IL-10 was measured by ELISA. The results showed that the level of IL-12 decreased and IL-10 increased after PBMNCs were incubated with MCF-7 cultural supernatant. 10 nmol/L neu-Fc strongly induced the cell proliferation. Compared with neu-Fc or GM-CSF or BCG treatment alone, neu-Fc in combination with GM-CSF and BCG significantly stimulated IL-12 production and inhibited IL-10 production (p < 0.01). It is concluded that the neu-Fc can stimulate the proliferation activity of PBMNCs. neu-Fc, GM-CSF and BCG costimulation efficiently induces Th1 immune response.


Subject(s)
Animals , BCG Vaccine , Allergy and Immunology , CHO Cells , Cricetinae , Cricetulus , Granulocyte-Macrophage Colony-Stimulating Factor , Allergy and Immunology , Humans , Interleukin-10 , Metabolism , Interleukin-12 , Metabolism , Rats , Recombinant Proteins , Allergy and Immunology , Th1 Cells , Allergy and Immunology , Th2 Cells , Allergy and Immunology
17.
Article in Chinese | WPRIM | ID: wpr-231903

ABSTRACT

<p><b>OBJECTIVE</b>To identify two differentiation-associated proteins induced by rhIL-6 in M1 mouse myeloid leukemia cells.</p><p><b>METHODS</b>Protein spots were excised from 2-D gels and digested in-gel with trypsin. The trypsin lysis products were first analyzed by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) through peptide mass fingerprinting and then performed peptide sequencing by nano-electrospray ionization mass spectrometry/mass spectrometry (nano-ESI-MS/MS). The database search was finished with the Mascot search engine (http://www.matrixscience.co.uk) using the data processed through MaxEnt3 and MasSeq.</p><p><b>RESULTS</b>The two proteins were not revealed by peptide mass fingerprint using MALDI-TOF-MS, while they were respectively identified as Destrin and Putative protein after the sequence of their trypic peptides were obtained by the nano-ESI-MS/MS techniques.</p><p><b>CONCLUSION</b>Nano-ESI-MS/MS technique can successfully identify the two differentiation-associated proteins induced by rhIL-6 and has great advantage in protein analysis.</p>


Subject(s)
Actin Depolymerizing Factors , Amino Acid Sequence , Animals , Apoptosis , Cell Transformation, Neoplastic , Destrin , Interleukin-6 , Pharmacology , Leukemia, Myeloid, Acute , Metabolism , Pathology , Mice , Microfilament Proteins , Molecular Sequence Data , Nanotechnology , Recombinant Proteins , Pharmacology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Methods , Tumor Suppressor Proteins
18.
Article in Chinese | WPRIM | ID: wpr-355653

ABSTRACT

Eukaryotic initiation factor 5A (eIF-5A) contains an unusual amino acid, hypusine, which is formed post-translationally. Although eIF-5A and its hypusine modification are essential for eukaryotic cell viability, the precise physiological function of it has remained elusive. The aim of the study is to investigate how hypusine formation modulate the proliferation, cell cycle and apoptosis in leukaemia cells. The effects of 1,7-diaminoheptane (DAH), a potent inhibitor of deoxyhypusine synthase, on proliferation and cell viability of leukemia cell lines (Mo7e, TF-1 and THP-1) and MCF-7 cells, were investigated. eIF-5A expression level was detected after cell synchronization. The results showed that inhibition of cell proliferation by DAH was in a concentration-dependent manner while apoptosis was also induced at the same time. Upon treatment of the cell lines with DAH, cell growth was inhibited. Cell cycle analysis showed that DAH induced cell growth arrest at the G(1)-S boundary of the cell cycle. In synchronized MCF-7 cells, the expression level of eIF-5A peaked at G(1) phase but very low at S and G(2)/M phases. It is concluded that hypusine formation of eIF-5A exits in the regulation of cell cycle and the results suggest that eIF-5A is involved in the expression of proteins regulating transition of G(1)-S phase of cell cycle.


Subject(s)
Cell Line, Tumor , Diamines , Pharmacology , G1 Phase , Physiology , Humans , Lysine , Metabolism , Peptide Initiation Factors , Physiology , RNA-Binding Proteins , S Phase , Physiology
19.
Article in Chinese | WPRIM | ID: wpr-258011

ABSTRACT

The ubiquitin-proteasome pathway is the principal mechanism for the degradation of short-lived proteins in eukaryotic cells. Recently, proteasome inhibitors have been shown to induce apoptosis in many kinds of human malignant cells. In this study, the mechanism of apoptosis induced by proteasome inhibitor in leukemic cells was examined. Evaluated by MTT assay, treatment of leukemic cells with Z-LLL-CHO, a reversible proteasome inhibitor, induced cell death in a dose-dependent manner. Appearance of the sub G(0)/G(1) fraction of cell cycle observed in flow cytometry assay suggested the induction of apoptosis, which was further proved by typical DNA ladder and morphological study. Western blot displayed the cleavage of bcl-2 into a shortened 22 kD fragment and the decrease in the levels of caspase-3 precursor. A highly sensitive colorimetric assay was employed and the elevation of caspase-3 activity was detected in both cell lines after treatment with Z-LLL-CHO. By comparison, these results showed that the leukemic cell line M-07e and KG-1a, which both express bcl-2 at a relative high level, had different susceptibility to undergo apoptosis induced by Z-LLL-CHO, which possibly due to their different levels of expression and activation of caspase-3 precursor, as well as their different degree of bcl-2 cleavage after treated by Z-LLL-CHO.

20.
Microbiology ; (12)1992.
Article in Chinese | WPRIM | ID: wpr-684453

ABSTRACT

The influence of medium variety、thermal treatment of the soil samples and inhibitor to the isolation effect of actinomycetes was studied by plate paint isolation methods.The results showed that: ①The isolation results of gauze No.1(GA) and straw decay substance agar(SDSA) might reflect basic station of soil actinomycetes. ②Thermal treatment (120℃?1.0h)in soils might promote actinomycetes spore sprouting and increase the quantity and variety of actinomycetes. The quantity and varieties of actinomycetes in the Soils that were treated with 120℃?1.0h were increased by 5.5%~54.9% and 12.5%~100% than the check of none-thermal treatment respectively. But thermal treatment had no effect on reduce the quantity of bacteria. ③The quantity of bacteria had significant reduction by adding 75?g/mL K 2Cr 2O 7 and 1~3?g/mL penicillin at same time to the isolation medium, and had little effect on the quantity and varieties of actinomycetes. Actinomycetes quantity (?10 6cfu/g) and variety of the treating of 75?g/mL K 2Cr 2O 7 and 1~3?g/mL penicillin in No.1 soil on SDSA medium had reductions by 0~58.8% and 0~18.2% compared with treating of 75?g/mL K 2Cr 2O 7, respectively. ④streptomycin might not been used for bacterteria inhabitor in the isolation of actinomycetes.

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