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1.
Chinese Journal of Anesthesiology ; (12): 1098-1102, 2022.
Article in Chinese | WPRIM | ID: wpr-957573

ABSTRACT

Objective:To evaluate the effect of hypothermia on the polarization of microglia and TLR4/NF-κB signaling pathway during oxygen-glucose deprivation/restoration (OGD/R).Methods:BV2 microglia were cultured in vitro and divided into 3 groups ( n=18 each) using the random number table method: control group (group C), group OGD/R and OGD/R plus hypothermia group (group OGD/R+ HT). Group C was cultured normally for 24 h. In group OGD/R, the cells were exposed to 5% CO 2-1% O 2 at 37 ℃ for 2 h in a glucose-free medium, followed by restoration of glucose and oxygen for 24 h. In group OGD/R+ HT, the high-glucose medium was replaced with a glucose-free medium, the cells were exposed to 5% CO 2-1% O 2 for 2 h in a 33 ℃ cryostat, followed by restoration of glucose and oxygen for 24 h. The cell survival rate was measured by CCK-8 assay.The expression of M1 microglia markers CD32 and iNOS protein and mRNA and M2 microglia markers CD206 and Arg-1 and mRNA was detected by immunofluorescence and real-time quantitative polymerase chain reaction.The expression of TLR-4 and NF-κB in cells was detected by Western blot, and the expression of TLR4 and NF-κB mRNA in cells was detected by real-time quantitative polymerase chain reaction.The concentrations of tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β) and interleukin 10 (IL-10) in the cell supernatant were detected by enzyme-linked immunosorbent assay. Results:Compared with group C, the cell survival rate was significantly decreased, the expression of CD32, iNOS, CD206, Arg-1, TLR4 and NF-κB protein and mRNA was up-regulated, and the concentrations of TNF-α, IL-1β and IL-10 in supernatant were increased in OGD/R and OGD/R+ HT groups ( P<0.05). Compared with group OGD/R, the cell survival rate was significantly increased, the expression of CD32, iNOS, TLR4 and NF-κB protein and mRNA was down-regulated, the expression of CD206 and Arg-1 protein and mRNA was up-regulated, the concentrations of TNF-α and IL-1β in supernatant were decreased, and the concentration of IL-10 was increased in group OGD/R+ HT ( P<0.05). Conclusions:Hypothermia can significantly inhibit microglia polarization toward M1 phenotype, increase microglia polarization toward M2 phenotype and inhibit the development of inflammatory responses during OGD/R, and the mechanism may be related to inhibition of TLR4/NF-κB signaling pathway.

2.
Article in Chinese | WPRIM | ID: wpr-706980

ABSTRACT

Objective To construct the evaluation indicator system for clinical scientific research on data quality; To provide a basis for assessing data quality of clinic scientific research.Methods Evaluation indicator system for clinic scientific research on data quality was established based on literature review and expert suggestion. A two- rounds Delphi process was conducted and the specialist participating level, concentration degree, coordination degree and authority coefficient were calculated to assess the indicator system.Results Two rounds of Delphi consultations was conducted by 30 experts that from different majors at the department of internal medicine or surgery, and both the effective recovery rates were 100%; the average authority coefficients of expert were 0.79 and 0.81, respectively. In the second round of survey, the final concentration degree ranged from 46.67% to 54.69% at full mark rate, and 4.23 to 4.36 at the mean values in 5 primary indicators. The CV for average 5 primary indicators ranged from 0.11 to 0.18. After 2 rounds of expert consultation, the final indicator system was consisted of 5 primary indicators (CRF construction, random control process, informed consent, data capture and data validation), 18 secondary indicators and 63 third-class indicators.Conclusion After two rounds of consultation, the opinions of all experts have been tending to be uniform, with high degree of coordination, and the indicator system can be used in assessment of data quality of clinic scientific research.

3.
Chin. med. j ; Chin. med. j;(24): 2958-2966, 2016.
Article in English | WPRIM | ID: wpr-230847

ABSTRACT

<p><b>BACKGROUND</b>Traditional Chinese medicine wogonin plays an important role in the treatment of leukemia. Recently, the application of drug-coated magnetic nanoparticles (MNPs) to increase water solubility of the drug and to enhance its chemotherapeutic efficiency has attracted much attention. Drugs coated with MNPs are becoming a promising way for better leukemia treatment. This study aimed to assess the possible molecular mechanisms of wogonin-coated MNP-Fe3O4 (Wog-MNPs-Fe3O4) as an antileukemia agent.</p><p><b>METHODS</b>After incubated for 48 h, the antiproliferative effects of MNPs, wogonin, or Wog-MNPs-Fe3O4on K562/A02 cells were determined by methyl thiazolyl tetrazolium (MTT) assay. The apoptotic rates of K562/A02 cells treated with either wogonin or Wog-MNPs-Fe3O4were determined by flow cytometer (FCM) assay. The cell cycle arrest in K562/A02 cells was determined by FCM assay. The elementary molecular mechanisms of these phenomena were explored by Western blot and reverse transcriptase polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>With cell viabilities ranging from 98.76% to 101.43%, MNP-Fe3O4was nontoxic to the cell line. Meanwhile, the wogonin and Wog-MNPs-Fe3O4had little effects on normal human embryonic lung fibroblast cells. The cell viabilities of the Wog-MNPs-Fe3O4group (28.64-68.36%) were significantly lower than those of the wogonin group (35.53-97.28%) in a dose-dependent manner in 48 h (P < 0.001). The apoptotic rate of K562/A02 cells was significantly improved in 50 μmol/L Wog-MNPs-Fe3O4group (34.28%) compared with that in 50 μmol/L wogonin group (23.46%; P< 0.001). Compared with those of the 25 and 50 μmol/L wogonin groups, the ratios of G0/G1-phase K562/A02 cells were significantly higher in the 25 and 50 μmol/L Wog-MNPs-Fe3O4groups (all P< 0.001). The mRNA and protein expression levels of the p21 and p27 in the K562/A02 cells were also significantly higher in the Wog-MNPs-Fe3O4group compared with those of the wogonin group (all P< 0.001).</p><p><b>CONCLUSIONS</b>This study demonstrated that MNPs were the effective drug delivery vehicles to deliver wogonin to the leukemia cells. Through increasing cells arrested at G0/G1-phase and inducing apoptosis of K562/A02 cells, MNPs could enhance the therapeutic effects of wogonin on leukemia cells. These findings indicated that MNPs loaded with wogonin could provide a promising way for better leukemia treatment.</p>


Subject(s)
Humans , Apoptosis , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Cell Survival , Drug Delivery Systems , Methods , Drug Resistance, Multiple , Drugs, Chinese Herbal , Chemistry , Pharmacology , Flavanones , Chemistry , Pharmacology , K562 Cells , Magnetics , Nanoparticles , Chemistry
4.
Chin. med. j ; Chin. med. j;(24): 741-746, 2012.
Article in English | WPRIM | ID: wpr-262533

ABSTRACT

<p><b>BACKGROUND</b>Dihydropyrimidine dehydrogenase (DPD), a key enzyme involved in the catabolism of 5-fluorouracil (5-FU), is the attractive candidate for pharmacogenetic research on efficacies and toxicities of 5-FU. The aim of this study is to explore the association between polymorphisms of dihydropyrimidine dehydrogenase gene (DPYD) and clinical outcomes of gastric cancer patients treated with fluorouracil-based adjuvant chemotherapy in the Chinese population.</p><p><b>METHODS</b>Three hundred and sixty-two patients with gastric cancer in the Chinese population were treated with fluorouracil-based adjuvant chemotherapy. The single nucleotide polymorphic genotypes of DPYD were determined by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) using DNA samples isolated from peripheral blood collected before treatment.</p><p><b>RESULTS</b>The average response rate for chemotherapy was 46.7%. A significantly different distribution of the rs1801159 (c2=8.76, P=0.012) genotypes was observed. Homozygous genotype rs1801159A/A was over-represented in responsive patients. Conversely, carriers of the rs1801159A/G genotype were prevalent in non-responsive patients. In the haplotype association analysis, there was significant difference in global haplotype distribution between the groups (c2=3.96, P=0.0465).</p><p><b>CONCLUSIONS</b>These results suggest that polymorphisms of rs1801159 in DPYD may be used as valuable predictors of the response to fluorouracil-based chemotherapy for gastric cancer patients in the Chinese population. Well-designed, comprehensive, and prospective studies on determining these polymorphisms of DPYD as predictive markers for gastric cancer in response to fluorouracil-based therapies are warranted.</p>


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Asian People , Chemotherapy, Adjuvant , Methods , Dihydrouracil Dehydrogenase (NADP) , Genetics , Fluorouracil , Therapeutic Uses , Genotype , Polymorphism, Single Nucleotide , Genetics , Stomach Neoplasms , Drug Therapy , Genetics , Treatment Outcome
5.
Article in Chinese | WPRIM | ID: wpr-332296

ABSTRACT

This study was purposed to detect single nucleotide polymorphisms (SNP) of 2 pharmacokinetics-related genes in K562 and K562/A02 cell lines. Leukemia cell line K562 and its resistant line K562/A02 were cultured, the genomic DNA was isolated by QIAamp DNA Blood Mini kit, primers were designed, the related DNA fragments were amplified by PCR. The SNP genotyping of mthfr gene rs1801131, rs1801133 and rs2274976 and dpyd gene rs1801159, rs1801160 and rs17376848 was performed by means of matrix assisted laser desorption ionization-time of flight mass spectrometry method (MALDI-TOFMS). The results showed that the genotype of mthfr gene locus 1801131 was AC, rs1801133 was CC, rs2274976 was GG, genotype of dpyd gene locus 1801159 was GG, rs1801160 was GG, rs17376848 was AA in both K562 and K562/A02 cell lines. It is concluded that the above-mentioned loci of mthfr and dpyd genes in K562 and K562/A02 cell lines are not expressed differently.


Subject(s)
Humans , DNA Mutational Analysis , DNA Primers , Dihydrouracil Dehydrogenase (NADP) , Genetics , Drug Resistance, Multiple , Genetics , Drug Resistance, Neoplasm , Genotype , K562 Cells , Methylenetetrahydrofolate Reductase (NADPH2) , Genetics , Polymorphism, Single Nucleotide
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