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1.
Article in Chinese | WPRIM | ID: wpr-879016

ABSTRACT

There is no consensus on the content, accumulation, transformation and content determination methods of phenolic acids in fresh Salvia miltiorrhiza. In order to find out the true content of phenolic acids in fresh S. miltiorrhiza, a variety of treatment me-thods were used in this study to prepare sample solution. The content changes of phenolic acids in S. miltiorrhiza samples with different dehydration rates were investigated during drying and shade drying processes. Polyphenol oxidase(PPO) of S. miltiorrhiza was extracted and purified by ammonium sulfate precipitation and dialysis to investigate the enzymatic properties. The content of rosmarinic acid, lithosperic acid and S. nolic acid B in S. miltiorrhiza was determined by UPLC. The results showed that the content of phenolic acids in fresh S. miltiorrhiza was highest when it was homogenized with 1 mol·L~(-1) HCl solution or 1 mol·L~(-1) HCl methanol solution. There was no significant difference in the content of phenolic acids in S. miltiorrhiza with different dehydration rates, indicating that there was no correlation between phenolic acid content and dehydration rate. The optimum pH of S. miltiorrhiza PPO was 7.6 and the optimum temperature was 40 ℃. With catechol as substrate, S. miltiorrhiza PPO had the enzymatic browning reaction which was in compliance with Michaelis equation, with Michaelis constant K_m of 0.12 mol·L~(-1) and V_(max) of 588.23 U·min~(-1). The inhibitory effect of citric acid, disodium ethylenediamine tetraacetate, ascorbic acid and sodium sulfite on S. miltiorrhiza PPO increased with the increase of inhibitor concentration, and sodium sulfite showed the strongest inhibitory effect. The present study proved that there were a large number of phenolic acids in fresh S. miltiorrhiza, which were the secondary metabolite of primitive accumulation during the growth of S. miltiorrhiza, rather than the induced product of postharvest drying and dehydration stress. This study has reference value and significance for the cultivation, harvest and processing of S. miltiorrhiza.


Subject(s)
Catechol Oxidase , Desiccation , Hydroxybenzoates , Plant Roots , Salvia miltiorrhiza
2.
Article in Chinese | WPRIM | ID: wpr-827191

ABSTRACT

OBJECTIVE@#To study the correlation of the expression alteration of Tim-3 with the T cell and B cell dysfunction in peripheral blood of multiple myeloma (MM) patients.@*METHODS@#30 patients diagnosed as MM from October 2016 to October 2018 were selected and enrolled in MM group, and 30 healthy persons whose sex and age was matched with the MM patients were selected and enrolled in healthy control group (HC). The blood samples from MM patients and HC were collected, and the peripheral blood mononuclear cells (PBMNC) were separated by density gradient centrifugation, then the serum was kept for further study. The ratios of CD3CD4Tim-3T cells, CD3CD8Tim-3T cells and the CD19+CD20-CD38+B cells were analysed by flow cytometry (FCM),and the concentration of T cell-related cytokines IFN-γ, TNF-αand B cell-related antibodies IgA, IgM and IgG were measured by ELISA. At the same time, the differences of the ratios of CCD3CD4Tim-3T, CD3CD8Tim-3T cells and plasmablast and the concentration of IFN-γ, TNF-α, IgA, IgM and IgG between the MM patient and HC were estimated, and the correlation of the ratio of CD3CD4Tim-3T, CD3CD8Tim-3T cells with the ratio of plasmablast and the concentration of IFN-γ, TNF-α, IgA, IgM and IgG in MM patients were analyzed.@*RESULTS@#The ratio of CD3CD4Tim-3T, CD3CD8Tim-3T cells increased in MM patients, while the ratio of CD19+CD20- CD38+B cells and the concentration of IFN-γ, TNF-α, IgA, IgM and IgG decreased in MM patients. And there was a negative correlation of the ratio of CD3CD4Tim-3T cells with CD19+CD20-CD38+B cells and the concentration of IFN-γ, IgA, IgM and IgG in MM patients, while the ratio of CD3CD8Tim-3T cells just negatively correlated with the concentration of TNF-α.@*CONCLUSION@#Expression of Tim-3 on CD4 and CD8 cells elevates in the peripheral blood of MM patients, which also correlates with the function suppression of T and B cells.


Subject(s)
B-Lymphocytes , CD8-Positive T-Lymphocytes , Hepatitis A Virus Cellular Receptor 2 , Metabolism , Humans , Leukocytes, Mononuclear , Multiple Myeloma
3.
Journal of Experimental Hematology ; (6): 1294-1300, 2018.
Article in Chinese | WPRIM | ID: wpr-689940

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the mutation rate and mutation characteristics of FBXW7 and NOTCH1 in adult T-ALL, and to study the effect of these 2 mutations on the clinical features and prognosis of patients with adult T-ALL.</p><p><b>METHODS</b>The mutations of FBXW7 and NOTCH1 in 106 adult T-ALL patients were determined by gene sequencing of FBXW7 and NOTCH1 genes. and the clinical characteristics and prognosis were compared.</p><p><b>RESULTS</b>Among the 106 cases of adult T-ALL, there were 21 cases (19.8%) of FBXW7 mutation, 66 cases (62.3%) of NOTCH1 mutation and 18 cases (17.0%) of FBXW7 / NOTCH1 double mutations. The 2-year cumulative overall survival rate and event-free survival rate of the patiants in FBXW7 / NOTCH1 double mutant group were lower than those without mutations (27.8% vs 70.3%)(P<0.01) and (16.7% vs 48.6%) ( P<0.01) respectivly, but the FBXW7 or NOTCH1 mutations had a little effect on the prognosis. The recurrence rate at mutant group was higher than that of the non - mutant group (77.8% vs 43.2%) (P<0.05).</p><p><b>CONCLUSION</b>FBXW7 or NOTCH1 mutations can not be as the prognostic factors, but the FBXW7 / NOTCH1 double mutations may indicate poor prognosis.</p>

4.
Chinese Traditional Patent Medicine ; (12): 1106-1109, 2018.
Article in Chinese | WPRIM | ID: wpr-710278

ABSTRACT

AIM To study the chemical constituents from the Stellaria dichotoma L.var.lanceolata Bge and their anti-inflammatory activities.METHODS The 95% ethnol extract from S.dichotoma was isolated and purified by silica,MCI,C18 prep-HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.Their anti-inflammatory activities were evaluated by LPS induced RAW264.7 inflammatory cell model.RESULTS Ten compounds were isolated and identified as 3-hydroxy-β-carboline (1),taraxacine A (2),1,2,3,4-tetrahydro-1,3,4-trioxo-β-carbo1ine (3),1-acetyl-β-carboline (4),arenarine A (5),arenarine B (6),diisobutyl-phthalate (7),dibutyl-phthalate (8),(z,z,z)-9,12,15-octadecatrienoic acid,methyl ester (9),tricin (10).Compounds 1-6 had stronger anti-inflammatory activities with the IC50 values of 4.79-9.34 μg/mL.CONCLUSION All the compounds are isolated from this plant for the first time.Their stronger anti-inflammatory activities are discovered for the first time.

5.
Article in English | WPRIM | ID: wpr-311363

ABSTRACT

We assessed the role of diabetes mellitus (DM) on treatment effects in drug-susceptible initial pulmonary tuberculosis (PTB) patients. A prospective study was conducted in eight provinces of China from October 2008 to December 2010. We enrolled 1,313 confirmed drug-susceptible initial PTB patients, and all subjects received the treatment regimen (2H3R3E3Z3/4H3R3) as recommended by the national guidelines. Of the 1,313 PTB patients, 157 (11.9%) had DM; these patients had more sputum smear-positive rates at the end of the second month [adjusted odds ratios (aOR) 2.829, 95% confidence intervals (CI) 1.783-4.490], and higher treatment failure (aOR 2.120, 95% CI 1.565-3.477) and death rates (aOR 1.536, 95% CI 1.011-2.628). DM was a contributing factor for culture-positive rates at the end of the second month and treatment failure and death of PTB patients, thus playing an unfavorable role in treatment effects of PTB.


Subject(s)
Antitubercular Agents , Therapeutic Uses , China , Epidemiology , Diabetes Mellitus , Epidemiology , Therapeutics , Female , Humans , Male , Mycobacterium tuberculosis , Tuberculosis, Pulmonary , Drug Therapy , Epidemiology , Microbiology
6.
Article in English | WPRIM | ID: wpr-296505

ABSTRACT

We assessed the incidence of adverse drug reactions (ADRs) with anti-TB medications and evaluated the risk factors for developing ADRs in previously treated tuberculosis patients in China. All patients received the first-line anti-TB regimen (2HREZS/6HRE) as recommended by the national guidelines. Clinical and laboratory evaluations were performed once a month. Out of the 354 participants, 262 (74.0%) experienced ADRs such as hyperuricemia (65.0%, 230/354), hepatotoxicity (6.2%, 22/354) and hearing disturbances (4.8%, 17/354). ADRs were significantly associated with diabetes mellitus [OR (95% CI): 15.5 (2.07-115.87)]; however, weight more than 50 kg [OR (95% CI): 0.41 (0.22-0.85)] was a protective factor for occurrence of ADRs. Hyperuricemia is the most common adverse event but, most patients with hyperuricemia showed increased tolerance for high uric acid levels. Low body weight and diabetes mellitus increased the risk of the occurrence of ADRs during anti-TB treatment.


Subject(s)
Adolescent , Adult , Antitubercular Agents , Therapeutic Uses , Diabetes Mellitus , Female , Humans , Male , Middle Aged , Prevalence , Retrospective Studies , Risk Factors , Tuberculosis, Pulmonary , Drug Therapy , Epidemiology , Young Adult
7.
Article in Chinese | WPRIM | ID: wpr-699890

ABSTRACT

Objective To develop an intelligent pneumatic tourniquet to reduce the complications due to manual operation.Methods The tourniquet was composed of two parts of tourniquet and electronic processor.The electronic processor involved in the pressure sensor,data processor,electronic display and alarm device.Moving average and frequency domain filtering algorithms were used to design signal processing.Results The tourniquet gained advantages over the traditional one in patient comfort,hemostatic efficacy and complications.Conclusion The tourniquet increases the safety while decreases the complications due to manual operation,and thus is worthy promoting clinically.

8.
Article in English | WPRIM | ID: wpr-296559

ABSTRACT

The objective of this prospective study of the risks of treatment failure in patients with drug-susceptible pulmonary tuberculosis (PTB) was to provide reference data to help develop a disease control strategy. Participants were recruited in eight provinces of China from October 2008 to December 2010. A total of 1447 patients with drug-susceptible PTB and older than 15 years of age were enrolled. Demographic characteristics, bacteriological test results, and patient outcome, i.e., cure or treatment failure were recorded and compared using the chi-square or Fisher's exact tests. Multivariate logistic regression was used to identify factors associated with risk of treatment failure. Of the 1447 patients who were enrolled, 1349 patients (93.2%) were successfully treated and 98 (6.8%) failed treatment. Failure was significantly associated with age 365 years [odds ratio (OR)=2.522, 95% confidence interval (CI): (1.097-5.801)], retreatment [OR=2.365, 95% CI: (1.276-4.381)], missed medicine [OR=1.836, 95% CI: (1.020-3.306)], treatment not observed [OR=1.879 95% CI: (1.105-3.195)], and positive culture result after the first [OR=1.971, 95% CI: (1.080-3.597)] and second month [OR=4.659, 95% CI: (2.590-8.382)]. The risk factors associated with treatment failure were age 365 years, retreatment, missed medication, treatment not observed, and positive culture at the end of month 1 or month 2. These risk factors should be monitored during treatment and interventions carried out to reduce or prevent treatment failure and optimize treatment success.


Subject(s)
Adolescent , Adult , Aged , Antitubercular Agents , Therapeutic Uses , China , Epidemiology , Female , Humans , Male , Middle Aged , Mycobacterium tuberculosis , Physiology , Prospective Studies , Retreatment , Risk Factors , Treatment Failure , Tuberculosis, Multidrug-Resistant , Drug Therapy , Epidemiology , Microbiology , Tuberculosis, Pulmonary , Drug Therapy , Epidemiology , Microbiology , Young Adult
9.
Article in English | WPRIM | ID: wpr-258839

ABSTRACT

Interleukin 8 (IL8) is an important chemokine that elicits host immune response against tuberculosis (TB). However, whether there is an association between IL8 gene polymorphism and TB susceptibility in the Chinese population is unknown. IL8 gene was amplified and sequenced to search for nucleotide polymorphisms among the Chinese population. Four single nucleotide polymorphisms (SNPs) were identified, selected, and analyzed in a cohort of 438 patients with TB and 536 healthy controls. Allelic, genotypic, and haplotypic analysis demonstrated that the distribution of the four IL8 SNPs between patients with TB and healthy controls was not significantly different (P>0.05). The four IL8 SNPs detected in this study were not associated with TB susceptibility in the Chinese population. Secretion of IL8 by peripheral blood cells was greatly stimulated upon exposure to Mycobacterium tuberculosis whole cell extract, but such enhanced secretion was not associated with the IL8 rs4073 alleles.


Subject(s)
Asian Continental Ancestry Group , Genetics , China , Genetic Predisposition to Disease , Humans , Interleukin-8 , Genetics , Polymorphism, Single Nucleotide , Tuberculosis , Genetics
10.
Article in Chinese | WPRIM | ID: wpr-341813

ABSTRACT

To study the effect of steaming and baking process on contents of alkaloids in Aconite Lateralis Radix (Fuzi), 13 alkaloids were analyzed by UPLC-MS/MS equipped with ESI ion source in MRM mode. In steaming process, the contents of diester-diterpenoid alkaloids decreased rapidly, the contents of monoester-diterpenoid alkaloids firstly increased, reached the peak at 40 min, and then deceased gradually. The contents of aconine alkaloids (mesaconine, aconine and hypaconine) increased all the time during processing, while the contents of fuziline, songorine, karacoline, salsolionl were stable or slightly decreased. In baking process, dynamic variations of alkaloids were different from that in the steaming process. Diester-diterpenoid alkaloids were degraded slightly slower than in steaming process. Monoester-diterpenoid alkaloids, aconine alkaloids and the total alkaloids had been destroyed at different degrees, their contents were significantly lower than the ones in steaming Fuzi at the same processing time. This experiment revealed the dynamic variations of alkaloids in the course of steaming and baking. Two processing methods which can both effectively remove the toxic ingredients and retain the active ingredients are simple and controllable, and are valuable for popularization and application.


Subject(s)
Aconitine , Aconitum , Chemistry , Alkaloids , Chromatography, High Pressure Liquid , Drug Stability , Drugs, Chinese Herbal , Hot Temperature , Plant Extracts , Steam , Tandem Mass Spectrometry , Time Factors
11.
Article in Chinese | WPRIM | ID: wpr-233733

ABSTRACT

<p><b>OBJECTIVE</b>To establish a colorectal cancer cell line with stable PRL-3 and CDH22 gene knock-down for investigating the role of PRL-3 and CDH22 genes in the carcinogenesis and metastasis of colorectal cancer.</p><p><b>METHODS</b>A recombinant lentiviral vector targeting CDH22 gene was obtained using the pENTRTM/U6 construct and pLenti6/BLOCK-iT (TM)-DEST vector. The recombinant lentivirus was harvested from 293FT cells cotransfected with the optimized ViraPower(TM) Packaging Mix and the pLenti6/BLOCK-iT(TM)-DEST expression construct. SW480/PRL-3- cells were infected with the recombinant lentivirus targeting CDH22, and SW480 cells with stable PRL-3 and CDH22 knock-down were screened by blasticidin selection. PRL-3 expression in the cells was determined by real-time RT-PCR. RESULTS The titer of the lentivirus for the second infection was 8 x 10(5) U/ml. Seventeen positive clones were selected, among which the Clone 1 exhibited substantially down-regulated CDH22 and PRL-3 mRNA expressions.</p><p><b>CONCLUSIONS</b>A human colorectal cancer cell line with stable PRL-3 and CDH22 gene knock-down has been established.</p>


Subject(s)
Cadherins , Genetics , Cell Line, Tumor , Clone Cells , Metabolism , Colorectal Neoplasms , Genetics , Pathology , Gene Expression Regulation, Neoplastic , Gene Knockdown Techniques , Methods , Humans , Lentivirus , Genetics , Neoplasm Proteins , Genetics , Protein Tyrosine Phosphatases , Genetics , RNA Interference , RNA, Messenger , Genetics , Metabolism
12.
Article in Chinese | WPRIM | ID: wpr-310352

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the personality traits and intensity dependence of auditory evoked potentials (AEPs) in chronic primary insomnia.</p><p><b>METHODS</b>Thirty-seven patients with chronic primary insomnia (insomnia group) and 44 healthy subjects (control group) were enrolled in the study. The AEPs were examined in insomnia and control groups; the personality traits were studied by Zuckerman's Sensation Seeking Scales (SSS) and Zuckerman-Kuhlman's Personality Questionnaire (ZKPQ); and the mood states by Plutchik-van Praag's Depression Inventory (PVP).</p><p><b>RESULT</b>The scores of neuroticism-anxiety and depression in insomnia group were significantly higher than those in control group (P<0.01); and the scores of impulsivity and aggression-hostility were also higher than those in control group (P<0.05); N1-P2 amplitude of AEP increased with stimulus intensity, which were significantly different in 70, 80, 90,100 dB (P<0.01). There were significant correlations between activity and N1 latency at 80 dB, activity and P2 latency at 100 dB (r=0.270, r=0.276, P<0.05); and between total scores of sensation seeking scale and N1-P2 amplitude (r=0.3746, r=0.35329, P<0.01) at 70 and 90 dB stimulus intensity in insomnia group. There were significant correlations among experience seeking and N1-P2 amplitude, experience seeking and slope rate (P<0.01) at 70, 80, 90, 100 dB stimulus intensity in insomnia group (r=0.539, r=0.3439, r=0.439, r=0.3278). There were significant correlations between sensation seeking of boredom susceptibility and slope rate (r=-0.282998, P<0.05) in insomnia group. There were significant correlations between thrill and adventure seeking and N1-P2 amplitude(r=0.2789, P<0.05) at 90 dB stimulus intensity in insomnia group; there were significant correlations between PVP and N1-P2 amplitude (r=-0.3434, r=-0.3158, P<0.05) at 70 dB and N1 latency at 80 dB in insomnia group.</p><p><b>CONCLUSION</b>Chronic primary insomnia sufferers have higher levels of neuroticism-anxiety, depression, aggression-hostility and impulsivity, and some are correlated with stimulus intensity dependence in AEP.</p>


Subject(s)
Adult , Case-Control Studies , Chronic Disease , Depression , Evoked Potentials, Auditory , Female , Humans , Male , Personality , Physiology , Sleep Initiation and Maintenance Disorders , Psychology , Surveys and Questionnaires , Young Adult
13.
Article in Chinese | WPRIM | ID: wpr-280161

ABSTRACT

<p><b>OBJECTIVE</b>To construct a lentiviral vector for RNA interference (RNAi) of PRL-3 gene and establish a human colon carcinoma cell line with PRL-3 gene knock-down.</p><p><b>METHODS</b>The plasmids were constructed expressing two different short hairpin RNAs (shRNA) targeting PRL-3 gene under control by the U6 promoter by lentiviral vector. An entry clone was generated in the pENTR/U6 vector. After identification with sequencing, a recombinant lentiviral vector was obtained using the pENTR/U6 construct and pLenti6/BLOCK-iT TM-DEST vector. The recombinant lentivirus was harvested from 293FT cells co-transfected with optimized ViraPower Packaging Mix and the pLenti6/BLOCK-iT -DEST expression construct. SW480 cells were infected with the recombinant lentivirus and the cells with stable PRL-3 gene knock-down were screened by blasticidin selection. PRL-3 expression was detected using real-time reverse transcription-polymerase chain reaction and Western blotting.</p><p><b>RESULTS</b>A recombinant lentiviral vector expressing shRNAs targeting PRL-3 gene was successfully established and confirmed by DNA sequencing. The recombinant lentivirus were harvested from 293FT cells with a viral titer of 6 x 10(5) pfu/L. Twelve clones of SW480 cells infected with the recombinant lentivirus were selected, and the Clone 1 exhibited significant knock-down of PRL-3 protein expression (72.9%).</p><p><b>CONCLUSION</b>The successful establishment of human colon carcinoma cell line with PRL-3 gene knock-down provide a basis for investigation of the role of PRL-3 gene in the metastasis of human colorectal carcinoma.</p>


Subject(s)
Base Sequence , Blotting, Western , Cell Line , Cell Line, Tumor , Colorectal Neoplasms , Genetics , Metabolism , Pathology , Gene Knockdown Techniques , Genetic Vectors , Genetics , Humans , Lentivirus , Genetics , Neoplasm Proteins , Genetics , Metabolism , Protein Tyrosine Phosphatases , Genetics , Metabolism , RNA Interference , Reverse Transcriptase Polymerase Chain Reaction , Transfection
14.
Article in Chinese | WPRIM | ID: wpr-280141

ABSTRACT

<p><b>OBJECTIVE</b>To construct a lentiviral expression vector for RNA interference of human CDH22 gene, and assess its gene silencing effect in colorectal cancer cells to provide a basis for investigating the role of CDH22 gene in the signaling pathway involved in human colorectal carcinoma metastasis.</p><p><b>METHODS</b>Human CDH22 gene short hairpin RNA (shRNA) sequence was designed using a software available on-line. After synthesis and annealing, the double-stranded oligonucleotides (dsOligoe) were cloned into the pENTR(TM)/U6 plasmid followed by sequence analysis. A positive clone was subcloned into pLenti6/BLOCK-iT(TM)-DEST vector and transformed into stb13 competent cells, with also verification by sequencing. The recombinant lentivirus was harvested from 293FT cells contransfected with the positive recombined plasmid and lentiviral packing materials. SW480 cells were infected with the recombinant lentivirus and the cells with stable CDH22 knock-down were screened by blasticidin selection. CDH22 expression in the cells was determined by real-time reverse transcription-polymerase chain reaction.</p><p><b>RESULTS</b>A recombinant lentiviral vector expressing shRNAs against CDH22 gene was obtained and confirmed by DNA sequencing. Fifteen clones of SW480 cells infected with the recombinant lentivirus were selected, and clone 11 exhibited substantial knock-down of CDH22 mRNA expression.</p><p><b>CONCLUSION</b>The lentiviral shRNA expression vector targeting human CDH22 gene capable of stable CDH22 gene knock-down in SW480 cells has been successfully constructed, which provides a basis for further study of the relationship between human colorectal carcinoma and CDH22 gene.</p>


Subject(s)
Base Sequence , Cadherins , Genetics , Cell Line, Tumor , Colorectal Neoplasms , Genetics , Metabolism , Pathology , Genetic Vectors , Humans , Lentivirus , Genetics , Molecular Sequence Data , RNA Interference , RNA, Small Interfering , Genetics , Transfection
15.
Neuroscience Bulletin ; (6): 180-184, 2007.
Article in English | WPRIM | ID: wpr-300979

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of nitric oxide (NO) on the expression of apelin receptor mRNA, as well as their correlation, in the caudate nucleus of rat.</p><p><b>METHODS</b>L-Arginine (L-Arg), N(G)-nitro-L-arginine methyl ester (L-NAME) and normal saline (NS) was separately microinjected into rat caudate nucleus. Expressions of neuronal NO synthase (nNOS) mRNA and apelin receptor mRNA were detected by RT-PCR at 4, 8, 12, 24 and 48 h after microinjection, and their correlation was determined.</p><p><b>RESULTS</b>The expressions of nNOS mRNA and apelin receptor mRNA were both significantly increased after microinjection of L-Arg, but significantly decreased after microinjection of L-NAME compared with the NS control group. The nNOS mRNA had a positive correlation with the expression of apelin receptor mRNA after microinjection of L-Arg and L-NAME.</p><p><b>CONCLUSION</b>The activity of NOS in the central nervous system, especially in the caudate nucleus, is one of the key factors for NO to exert many kinds of biological actions, such as modulation of central pain, as a neurotransmitter. The neurobiological action of NO in rat caudate nucleus may be associated with apelin receptors.</p>


Subject(s)
Animals , Apelin Receptors , Arginine , Pharmacology , Caudate Nucleus , Metabolism , Enzyme Inhibitors , Pharmacology , Gene Expression Regulation , Physiology , Male , NG-Nitroarginine Methyl Ester , Pharmacology , Nitric Oxide , Physiology , Nitric Oxide Synthase Type I , Metabolism , RNA, Messenger , Rats , Rats, Wistar , Receptors, G-Protein-Coupled , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Methods
16.
Article in Chinese | WPRIM | ID: wpr-268122

ABSTRACT

<p><b>OBJECTIVE</b>To identify the region in PRL-3 gene promoter where the transcriptional factor Snail can bind.</p><p><b>METHODS</b>PRL-3 promoter and the possible binding sites of the transcription factors were analyzed by bioinformatical methods. Chromatin immunoprecipitation and PCR were performed using the antibody specific for Snail to verify the binding of Snail to PRL-3 promoter.</p><p><b>RESULTS</b>According to the prediction by TRED, a promoter prediction software, PRL-3 gene promoter was located between -700 bp to 299 bp of PRL-3 gene. Many possible transcription factor binding sites such as for Snail, n-MYC, ARNT, E74A, NF-kappaB, NRF-2 and AML-1 were predicted by Consite, a promoter analysis web system. Interestingly, a 5'-CACCTG-3' core sequence and other related sequences of Snail binding sites were found in the promoter region of PRL-3 genes by Consite software. Two regions in PRL-3 promoter were validated to allow binding of Snail by chromatin immunoprecipitation analysis of SW480 cells.</p><p><b>CONCLUSIONS</b>Snail regulates the activity of PRL-3 gene by binding to the promoter of PRL-3 gene in SW480 cells.</p>


Subject(s)
Base Sequence , Binding Sites , Cell Line, Tumor , Computational Biology , Humans , Molecular Sequence Data , Neoplasm Proteins , Metabolism , Promoter Regions, Genetic , Protein Tyrosine Phosphatases , Metabolism , Snail Family Transcription Factors , Software , Transcription Factors , Metabolism
17.
Article in Chinese | WPRIM | ID: wpr-268059

ABSTRACT

<p><b>OBJECTIVE</b>To obtain the entire coding sequence of human PRL-3 gene and construct its prokaryotic expression vector.</p><p><b>METHODS</b>With total RNA extracted from the human colorectal carcinoma cell line SW480 as the template, PRL-3 gene was amplified by RT-PCR with primers designed according to the published sequence of GenBank, and the product was inserted into pGEM-T Easy vector. The recombinant plasmid pGEM-T-PRL-3 was identified by restriction endonuclease analysis and DNA sequencing. After digestion with restriction endonuclease, PRL-3 gene was cloned into the multicloning sites of the prokaryotic expression vector pGEX-4T-1.</p><p><b>RESULTS AND CONCLUSION</b>The entire coding region of human PRL-3 gene was cloned, and the recombinant pGEX-4T-1-PRL-3 vector was successfully constructed and expressed, which may provide the basis for further study of the relationship between human colorectal carcinoma and PRL-3 gene.</p>


Subject(s)
Base Sequence , Cell Line, Tumor , Cloning, Molecular , DNA, Complementary , Chemistry , Genetics , Electrophoresis, Polyacrylamide Gel , Escherichia coli , Genetics , Genetic Vectors , Genetics , Humans , Molecular Sequence Data , Neoplasm Proteins , Genetics , Metabolism , Protein Tyrosine Phosphatases , Genetics , Metabolism , Recombinant Fusion Proteins , Genetics , Metabolism , Sequence Analysis, DNA
18.
Chinese Journal of Epidemiology ; (12): 665-668, 2005.
Article in Chinese | WPRIM | ID: wpr-331811

ABSTRACT

<p><b>OBJECTIVE</b>To study the association between reduced folate carrier gene (RFC1 A80G) polymorphism and the risk for child with neural tube defects (NTDs), and to provide epidemiological evidence for the existence of NTDs genetic marker.</p><p><b>METHODS</b>RFC1 (A80G) genotypes were detected using RFLP-PCR for blood DNA of 104 families with NTDs-affected children and 100 control families with no history of child-affected birth defects. Case-control study and transmission/disequilibrium test(TDT) for the RFC1 genotype of NTDs pedigree were carried out.</p><p><b>RESULTS</b>The G allele frequency of children with NTDs was higher than that of controls when compared to A allele( OR = 1. 64, 95% CI :1.08-2.49). The offspring of the GG genotype were associated with a 2.56-fold increased risk of NTDs when compared to the AA genotype (OR = 2.56, 95% CI: 1.04-6.36) in our study population. There was evidence of association between G allele and the risk of parent having a child with NTDs (OR = 1.56, 95% CI: 1.07-2.28) in the TDT analysis.</p><p><b>CONCLUSION</b>Our findings indicated that there was potential association between offspring RFC1 GG genotype and the risk of NTDs, and the G allele was a possible susceptible gene marker for an increased NTDs risk in the Chinese population.</p>


Subject(s)
Case-Control Studies , Child , Child, Preschool , Female , Gene Frequency , Genetic Predisposition to Disease , Genotype , Humans , Infant , Male , Membrane Transport Proteins , Genetics , Neural Tube Defects , Genetics , Parents , Polymorphism, Genetic , Reduced Folate Carrier Protein
19.
Acta Pharmaceutica Sinica ; (12): 677-679, 2003.
Article in Chinese | WPRIM | ID: wpr-266591

ABSTRACT

<p><b>AIM</b>To study the chemical constituents of the stems of Opuntia vulgaris Mill(Cactaceae).</p><p><b>METHODS</b>The compounds of Opuntia vulgaris were isolated by chromatography of Amberlite Dowex 50 and silica gel, and identified by means of UV, IR, MS, 1D and 2D NMR.</p><p><b>RESULTS</b>Three compounds were isolated and identified as: opuntin B(I), 4-hydroxyproline(II) and tyrosine(III).</p><p><b>CONCLUSION</b>Compound I is a new alkaloid.</p>


Subject(s)
Hydroxyproline , Chemistry , Maleimides , Chemistry , Molecular Conformation , Molecular Structure , Opuntia , Chemistry , Phenols , Chemistry , Plants, Medicinal , Chemistry , Tyrosine , Chemistry
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