2.The effects of anti-cancer drug NSC319726 on the gene expression of ovarian cancer
Yanhong DENG ; Xiu LE ; Wenchao GAO
Practical Oncology Journal 2016;30(4):332-338
Objective To study the effects of anti -cancer drug NSC319726 on the gene expression of ovarian cancer.Methods The data that NSC319726 acted on the ovarian cancer cell lines TOV112D was down-loaded from the GEO database and then the bioinformatical analysis was conducted .Results NSC319726 in-creased 246 genes including MMP3,CXCR4 et al and decreased 198 genes such as PBX1 in ovarian cancer cell lines.GO analysis indicated that six GO items were related to metabolism .Pathway analysis revealed that NSC319726 could affect the circadian clock and MMP signaling pathway .Conclusion NSC319726 could affect the expression of multiple functional genes in ovarian cancer cell lines and might be a potential drug targeting to the precursor-B cell acute lymphoblastic leukemia .Meanwhile , it should be noted that NSC 319726 may have effects on the tumor metastasis and human sleep .Therefore,further research needs to be conducted before the clinical use .
3.Establishment of prokaryotic expression and optimization ox expression conditions of Eleutherococcus senticosus P450 gene.
Peng WU ; Le-shan XIU ; Fei-fei LI ; Zhao-bin XING
China Journal of Chinese Materia Medica 2015;40(7):1274-1277
According to the sequence of P450 cDNA of Eleutherococcus senticosus, specific primers were designed. Frokaryotic ex pression vector pET30a-P450 was constructed and the prokaryotic expression conditions were optimized. Results showed that the BL21 after being transformed with the recombinant expression vector accumulated the high amount of recombinant protein. SDS-PAGE analysis showed that the recombinant protein was about 53 kDa. The recombinant accumulated the highest amount of recombinant protein af ter IPTG (1 mmol x L(-1)) at 27-37 degrees C for 24 h. Consequently P450 gene of E. senticosus could be expressed successfully by prokaryotic expression vector pET30a-P450. Induction temperature, IPTG concentration, medium type and amount of induction time could all influence the expression of target protein, but the impact strength was different.
Cloning, Molecular
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Cytochrome P-450 Enzyme System
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genetics
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metabolism
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Eleutherococcus
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enzymology
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genetics
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Escherichia coli
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genetics
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metabolism
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Gene Expression
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Genetic Vectors
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genetics
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metabolism
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Plant Proteins
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genetics
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metabolism
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Plasmids
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genetics
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metabolism
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Recombinant Proteins
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genetics
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metabolism
4.Evaluation of Risk Factors for Arytenoid Dislocation after Endotracheal Intubation:a Retrospective Case-control Study
Shen LE ; Wang WU-TAO ; Yu XUE-RONG ; Zhang XIU-HUA ; Huang YU-GUANG
Chinese Medical Sciences Journal 2014;(4):221-224
Objective To investigate the risk factors for postoperative arytenoid dislocation.
Methods From September 2003 to August 2013, the records of 16 patients with a history of postoperative arytenoid dislocation were reviewed. Patients matched in terms of date and type of procedures were chosen as the controls (n=16). Recorded data for all patients were demographics, smoking status, alcoholic status, preoperative physical status, airway evaluation, intubation procedures, preoperative laboratory test results, anesthetic consumption and intensive care unit stay. For arytenoid dislocation cases, we further analyzed the incidences of the left and right arytenoid dislocation, and the outcomes of surgical repair and conservative treatment. Categorical variables were presented as frequencies and percentages, and were compared using the chi-squared test. Continuous variables were expressed as means±SD and compared using the Student’s unpaired t-test. To determine the predictors of arytenoid dislocation, a logistic regression model was used for multivariate analysis.
Results Sixteen patients with postoperative arytenoid dislocation were enrolled, with a median age of 52 years. Most postoperative arytenoid dislocation patients (15/16, 93.75%) received surgical repair, except one patient who recovered after conservative treatment. None of the postoperative arytenoid dislocation patients were smokers. Red blood cell (P=0.044) and hemoglobin (P=0.031) levels were significantly lower among arytenoid dislocation cases compared with the controls.
Conclusions Non-smoking and anemic patients may be susceptible to postoperative arytenoid dislocation. However, neither of them was independent risk factor for postoperative arytenoid dislocation.
5.Corneal epithelial remodeling and its relationship with diopter after SMILE for moderate and high myopia
Juan Shu LIU ; Jie HOU ; Le Le ZHANG ; Lin Yu LEI ; Yun Xiu ZHENG
Recent Advances in Ophthalmology 2017;37(11):1060-1063
Objective To evaluate the changes in epithelial thickness profile and its relationship with diopter following small incision lenticule extraction (SMILE) for moderate and high myopia.Methods Together 46 myopia or myopic astigmatism (92 eyes) who underwent SMILE were included under the informed consent from January 2016 to March 2017 and were decided into 2 groups according to the diopter:moderate myopia group (58 eyes)and high myopia group (34 eyes).Epithelial thickness profile was measured using spectral-domain optical coherence tomography at different corneal zones (0-2 mm,> 2-5 mm and > 5-6 mm cornea) preoperatively before surgery and 1 month and 6 months after surgery for observing the changes in epithelial thickness and its correction with diopter.Results The mean epithelial thickness in the central zone was (55.68 ± 3.61) μm before surgery,and,6 months after surgery,it was thickened by (3.85-±3.99),(3.46 ±3.29) and (2.85 ±3.18) μm in the 0-2 mm,>2-5 mum and >5-6 mm cornea respectively,and the difference was statistically significant among the three zones (P < 0.01).After surgery for 6 months,the epithelial thickness in the high myopia group was thickened more obviously compared with the moderate myopia group (t =1.440,P =-0.047).And no correlation was found between changes in the epithelial thickness at 0-2 mm cornea and diopter after surgery(moderate myopia group:r =0.219,P=0.633;high myopia group:r =0.197,P =0.585).Conclusion Significant epithelial thickening was observed after SMILE,presenting the thickened epithelium.The higher the diopter was,the more thickened the epithelium was.The epithelial changes does not appear to affect the diopter after SMILE.
6.Myricetin inhibits interferon-γ-induced programmed death ligand-1 and indoleamine 2, 3-dioxygenase 1 expression in lung cancer cells
Yu-Chi CHEN ; Xin-Ling HE ; Lu QI ; Wei SHI ; Luo-Wei YUAN ; Mu-Yang HUANG ; Yu-Lian XU ; Xiu-Ping CHEN ; Le-Le ZHANG ; Jin-Jian LU
Chinese Journal of Pharmacology and Toxicology 2021;35(10):761-761
OBJECTIVE Programmed death ligand-1 (PD-L1) and indoleamine 2, 3-dioxygenase 1 (IDO1) are immune checkpoints which can be induced by interferon-γ(IFN-γ) in the tumor microenvironment, leading to immune escape of tumors. Myricetin (MY) is a flavonoid distributed in many edible and medicinal plants. The aim of this study is to clarify the effect and the mechanism of MY on inhibiting IFN-γ-induced PD-L1 and IDO1 in lung cancer cells. METHODS Expressions of PD-L1 and major histocompatibility complex-I (MHC-I) were evaluated by flow cytometry and Western blotting, and the expression of IDO1 was measured by Western blotting. qRT-PCR was used to detect their mRNA levels. The function of T cells was evaluated using a co-culture system consist of lung cancer cells and the Jurkat-PD-1 T cell line that overexpressing PD-1. Molecular docking analysis, Western blotting and immunofluorescence were used for mechanism study. RESULTS MY potently inhibited IFN-γ-induced PD-L1 and IDO1 expression in human lung cancer cells, while didn't show obvious effect on the expression of MHC-I. In addition, MY restored the survival, proliferation, CD69 expression and interleukin-2 (IL-2) secretion of Jurkat-PD-1 T cells suppressed by IFN-γ-treated lung cancer cells in the co-culture system. Mechanistically, IFN-γ up-regulated PD-L1 and IDO1 at the transcriptional level through the JAK-STAT-IRF1 axis, which was targeted and inhibited by MY. CONCLUSION Our research revealed a new insight into the anti-tumor effects of MY which inhibited IFN-γ-induced PD-L1 and IDO1 expression, supporting the potential of MY in anti-tumor immunotherapy.
7.Study of single cell PCR for HLA typing.
Dong LI ; Le-ling ZHANG ; Xiu-li JU ; Huai-shui HOU ; Qing SHI ; Bai-jun SHEN
Chinese Journal of Hematology 2007;28(5):308-311
OBJECTIVETo apply the single cell nested multiplex polymerase chain reaction (PCR) to HLA typing, and analyze the influence factors on the amplification results.
METHODSSingle cell DNA templates were prepared with different methods. The exon 2, 3 and intron 2 of HLA-A, B, and exon 2 of DRBI were amplified using multiplex PCR. The second round of SSP-PCR HLA typing was carried out according to the large scale routine HLA typing results.
RESULTSEnzyme lysis method was the most efficient procedure for preparing the single cell DNA template, with a success rate (SR) of 93.3%, while the SRs of alkali lysis and freezing-thaw lysis methods were 83.3% and 73.3%, respectively. The second round amplification using enzyme lysis and SSP-PCR in 20 samples obtained a 95% success rate and a 15% allele drop out rate. The time for performing the whole procedure was less than 6 hours.
CONCLUSIONThe modified nested multiplex PCR technique is efficient for single cell HLA typing and might be applied to clinical preimplantation genetic diagnosis.
Histocompatibility Testing ; methods ; Humans ; Polymerase Chain Reaction ; methods
8.Analysis on codon usage of chloroplast genome of Eleutherococcus senticosus.
Zhao-Bin XING ; Lei CAO ; Mi ZHOU ; Le-Shan XIU
China Journal of Chinese Materia Medica 2013;38(5):661-665
OBJECTIVETo analyze the codon usage of chloroplast genome and the influencing factor in Eleutherococcus senticosus.
METHODCodon of 52 genes, which were selected from the chloroplast genome sequence of E. senticosus, was multivariate statistical and correspondence analyzed using CodonW and SPSS software.
RESULTGC content at the three position of codons by turns was 46.46%, 38.26%, 29.88%, whereas GC1 and GC2 had a significant correlation coefficient (P < 0.01). The correlation coefficient with GC12, and GC3 was 0.205 and was not significant correlated. There were 30 codons which relative synonymous codon usage was greater than 1 and 29 codons end with A and T. In the corresponding analysis, the first axis shows 10.35% variation. And there was significant correlation coefficient between ENC and GC3. The correlation coefficients with GC3 and ENC were -0.288 and 0.353, respectively. We defined 16 codons from 16 amino acids as the major preference codons in chloroplast genome of E. senticosus.
CONCLUSIONThe third positions for all codon are preferred to ending with A and T. The codon usage bias is formed under effect of mutation and selection, as well as other factors. But the selection will have a far greater impact than others.
Amino Acids ; genetics ; Chloroplasts ; genetics ; Codon ; genetics ; Eleutherococcus ; genetics ; Genome, Plant ; genetics ; Genomics ; Multivariate Analysis ; Mutation
9.Merkel cell carcinoma on left thigh: a case report.
Hui-ling WU ; Li-jun ZHENG ; Yu-sheng YU ; Xiu-di YE ; Shu-jun LE
Journal of Zhejiang University. Medical sciences 2006;35(4):463-464
Aged
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Carcinoma, Merkel Cell
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diagnosis
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surgery
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Female
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Humans
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Skin Neoplasms
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diagnosis
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surgery
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Thigh