1.The expression and significance of Survivin mRNA in human prostate cancer
Xin YANG ; Qun XIE ; Yadong ZHANG
Chinese Journal of Primary Medicine and Pharmacy 2008;15(11):1831-1832
Objective To evaluate the expression of Srvivin mRNA in the occurrence and progress of prostate cancer.Methods The inhibitor of apoptosis gene Survivin mRNA in 45 prostate cancer cases and 30 benign prostatic hyperplasia(BPH) cases were detected by hybridization in situ technique.Results The average optical density of prostate cancer groups(0.4232±0.0085)was signifi cantly higher than BPH groups (0.3303±0.0834) (P=0.001 ).Among the different pathological grades [G1(0.3401±0.0474), G2 (0.4270±0.0074), G3 (0.4560±0.0883)],the difference was statistically significant in the expression of Survivin mRNA (P= 0.004).It was found that there was positive correlation between the transfer of lymph (0.4557±0.7921)and the expression of Survivin mRNA in prostate cancer(P=0.005 ).Conclusion Survivin mRNA is overexpression in prostate cancer.It presumed that Survivin participate in genesis,development of prostate cancer and closely correlated with prognosis.
2.Experimental study on bone formation with a novel borosilicate glass combined with platelet-rich plasma
Yadong ZHANG ; Gang WANG ; Xin LIU ; Wenchan HUANG ; Changqing ZHANG ; Zongpin XIE ; Shengbao CHEN
Chinese Journal of Orthopaedics 2010;30(8):789-795
Objective To evaluate the osteogenesis of a novel borosilicate glass materials combined with platelet-rich plasma in repairing segmental bone defects. Methods 36 New Zealand white rabbits which bilateral radius were resected into l.5cm bone defect, were divided into 4 groups averagely depending on implanted materials: group A (one side: D-Alk- 1B, another side: D-Alk- 1B +PRP), group B (one side:D-Alk- 1B +PRP, another side: β-TCP); group C (one side: β-TCP, another side: experimental bone defect),and group D(one side:D-Alk-1 B,another side :experimental bone defect). The specimens were examined after 4, 8,12 weeks; the osteogenesis was evaluated through gross observation, X-ray radiograph,histological examination,scanning electron microscope and Micro-CT. Results There were similar results about gross observation,X-ray radiograph ,histological examination. After 4, 8, 12 weeks ,D-Alk-1B materials, β-TCP and D-Alk-1 B + PRP group had better osteogenesis ability than the experimental control group (P <0.05); D-Alk-lB + PRP had the best performance, better than D-Alk-1B and β-TCP (P<0.05); D-Alk-1B were similar to β-TCP (P>0.05). D-Alk-1 B materials degradated faster than β-TCP materials, and the porous structure of the materials disappeared after degradation. D-Alk-1B materials intergrated with host's bone was better than β-TCP materials. Conclusion D-Alk-1B material have good biological activity, histocompatibility and biodegradation and simiar presence of bone formation compared withβ-TCP in the aspect of repairing the segmental bone defect, the combination of PRP and D-Alk-1 B strengthened osteogenesis in vivo.
3.Effects of short hairpin RNA-mediated semaphorin 5A gene silencing on proliferation, metastasis and invasion ;of malignant melanoma cell line A375
Li ZHANG ; Yadong LI ; Chen CHEN ; Lingjia LI ; Yuyan XIE ; Tongyun LIU ; Wei CUN
Chinese Journal of Dermatology 2016;49(8):573-577
Objective To study the effects of semaphorin 5A (SEMA5A) gene silencing by lentivirus?mediated short hairpin RNA(shRNA)on biological activity of malignant melanoma cell line A375. Methods Two pairs of interference sequences for SEMA5A gene(shRNA1 and A375?shRNA2)and a pair of control interference sequences were designed to build lentiviral vectors, which were then transfected into HEK293T cells to gain lentivirus. A375 cells were divided into three groups:experimental group(A375?shRNA1 and A375?shRNA2 cells)transfected with the lentivirus containing shRNA1 or shRNA2, negative control group (A375?con cells) transfected with that containing the control shRNA, and blank control group(A375 cells)receiving no transfection. The A375 cells with stable knockdown of SEMA5A gene expression were screened by puromycin. Subsequently, reverse transcription?PCR and Western?blot analysis were performed to detect mRNA and protein expressions of Semaphorin 5A in these cells, and methyl thiazolyl tetrazolium(MTT)assay was applied to evaluate the growth of cells. The scratch assay and invasion assay were conducted to estimate migration and invasion ability of cells. Results The lentivirus containing the SEMA5A?targeting shRNAs or control shRNA was successfully transfected into A375 cells, and stably transfected cells were gained after puromycin selection. The expressions of semaphorin 5A mRNA and protein in the A375?shRNA2 cells were significantly reduced compared with those in the A375?con and A375 cells(all P < 0.05). MTT assay showed that the growth of A375?shRNA2 cells was significantly slower than that of A375?con and A375 cells(both P<0.05), while there was no significant difference in the growth rate between A375?con and A375 cells(P>0.05). The scratch assay showed that there was no obvious cell migration into the scratch in the experiment group, whereas the scratch was almost covered by cells in the negative control group and blank control group. The invasion assay showed that the number of A375?shRNA2 cells passing through the Transwell chamber was significantly smaller than that of A375 and A375?con cells(both P<0.05), while there was no significant difference between that of A375 and A375?con cells(P > 0.05). Conclusion The silencing of SEMA5A gene by lentivirus?mediated shRNA could effectively down?regulate the expression of semaphorin 5A, and inhibit the growth, invasion and migration of A375 cells.
4.Directed evolution of aflatoxin detoxifzyme in vitro by error-prone PCR.
Sai ZHANG ; Keke XING ; Yadong HU ; Chunfang XIE ; Daling LIU ; Dongsheng YAO
Chinese Journal of Biotechnology 2011;27(7):1100-1108
The experiment was conducted by directed evolution strategy (error-prone PCR) to improve the activity of aflatoxin detoxifzyme with the high-throughput horse radish peroxidas and recessive brilliant green (HRP-RBG) screening system. We built up a mutant library to the order of 10(4). Two rounds of EP-PCR and HRP-RBG screening were used to obtain three optimum mutant strains A1773, A1476 and A2863. We found that mutant A1773 had upper temperature tolerance of 70 degrees C and that its enzyme activity was 6.5 times higher than that of the parent strain. Mutant strains A1476 worked well at pH 4.0 and its enzyme activity was 21 times higher than that of the parent strain. Mutant A2863 worked well at pH 4.0 and pH 7.5, and its enzyme activity was 12.6 times higher than that of the parent strain. With DNA sequencing we found that mutant A1773 revealed two amino acid substitutions, Glu127Lys and Gln613Arg. Mutant A1476 revealed four amino acid substitutions: Ser46Pro, Lys221Gln, Ile307Leu and Asn471lle. Mutant A2863 revealed four amino acid substitutions: Gly73Ser, Ile307Leu, Va1596Ala and Gln613Arg. The results provided a useful illustration for the deep understanding of the relationship between the function and structure of aflatoxin detoxifzyme.
Aflatoxin B1
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antagonists & inhibitors
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chemistry
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Amino Acid Substitution
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Directed Molecular Evolution
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Enzyme Activation
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Enzyme Stability
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Multienzyme Complexes
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genetics
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metabolism
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Mutant Proteins
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genetics
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metabolism
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Point Mutation
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Polymerase Chain Reaction
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methods
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Protein Engineering
5.Differential Hrd1 Expression and B-Cell Accumulation in Eosinophilic and Non-eosinophilic Chronic Rhinosinusitis With Nasal Polyps.
Kun CHEN ; Miaomiao HAN ; Mengyao TANG ; Yadong XIE ; Yuting LAI ; Xianting HU ; Jia ZHANG ; Jun YANG ; Huabin LI
Allergy, Asthma & Immunology Research 2018;10(6):698-715
PURPOSE: Hrd1 has recently emerged as a critical regulator of B-cells in autoimmune diseases. However, its role in the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP) remains largely unexplored. This study aimed to examine Hrd1 expression and B-cell accumulation and their possible roles in CRSwNP. METHODS: Quantitative real-time polymerase chain reaction, immunohistochemistry, enzyme-linked immunosorbent assay and Western blotting were used to assess gene and protein expression in nasal tissue extracts. Cells isolated from nasal tissues and peripheral blood mononuclear cells were characterized by flow cytometry. Local antibody production was measured in tissue extracts with a Bio-Plex assay. Additionally, changes in Hrd1 expression in response to specific inflammatory stimuli were measured in cultured dispersed polyp cells. RESULTS: Nasal polyps (NPs) from patients with eosinophilic CRSwNP (ECRS) had increased levels of Hrd1, B-cells and plasma cells compared with NPs from patients with non-eosinophilic CRSwNP (non-ECRS) or other control subjects (P < 0.05). The average Hrd1 levels in B-cells in NPs from ECRS patients were significantly higher than those from non-ECRS patients and control subjects (P < 0.05). NPs also contained significantly increased levels of several antibody isotypes compared with normal controls (P < 0.05). Interestingly, Hrd1 expression in cultured polyp cells from ECRS patients, but not non-ECRS patients, was significantly increased by interleukin-1β, lipopolysaccharide and Poly(I:C) stimulation, and inhibited by dexamethasone treatment (P < 0.05). CONCLUSIONS: Differential Hrd1 expression and B-cell accumulation between the ECRS and non-ECRS subsets suggests that they can exhibit distinct pathogenic mechanisms and play important roles in NP.
Antibody Formation
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Autoimmune Diseases
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B-Lymphocytes*
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Blotting, Western
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Dexamethasone
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Enzyme-Linked Immunosorbent Assay
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Eosinophils*
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Flow Cytometry
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Humans
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Immunity, Innate
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Immunohistochemistry
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Nasal Polyps*
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Plasma Cells
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Polyps
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Real-Time Polymerase Chain Reaction
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Tissue Extracts
6.Sepsis due to ureteral stones in a patient with uremia: a case report
Junlin XU ; Jie LIN ; Jiangling XIE ; Yadong TAN ; Renju LI ; Hui YANG
Chinese Journal of Urology 2023;44(5):392-393
Urosepsis caused by upper urinary tract stone obstruction is a common critically disease in urology.However, it rarely occurs in the patient who underwent a dialysis with uremia.We report a patient who underwent an implantation of ureteral stent to control the infection, and we saved the patient with perinephric hematoma following the surgery. We removed the stones in the left ureteral through a flexible ureteroscope two month later.The hematoma was completely absorbed 6 months after the implantation of ureteral stent.
7.ADE signal mining and analysis of axitinib based on FAERS database
Runan XIA ; Ting YING ; Hai LIANG ; Jiahui DAI ; Yadong WANG ; Xuefeng XIE
China Pharmacy 2023;34(23):2896-2900
OBJECTIVE To provide references for the clinical safe use of axitinib. METHODS Adverse drug event (ADE) data for axitinib were collected from the US FDA Adverse Event Reporting System (FAERS) database from the first quarter of 2012 to the fourth quarter of 2022. The data were mined and analyzed by utilizing the ratio-of-reporting-ratio (ROR) method and comprehensive standard method of the United Kingdom’s Medicines and Healthcare Products Regulatory Agency (MHRA) of proportional imbalance measurement. RESULTS A total of 13 962 reports of axitinib-related ADEs were obtained, with patients’ age concentrated in 65-85 years (43.25%), gender predominantly male (65.23%), country of reporting predominantly US (60.01%), and serious ADE outcomes mostly hospitalization or prolonged hospitalization (31.51%). A total of 172 ADE risk signals were detected, involving 18 system and organ classifications (SOC), mainly systemic diseases and various reactions at the site of administration (3 749 cases, 30.84%) and gastrointestinal system diseases (2 067 cases, 17.00%). ADE risk signals that occurred more frequently were generally consistent with the drug instruction, such as diarrhea, fatigue, and hypertension; new ADE risk signals requiring clinical attention were death, immune-mediated nephritis, and PT signals contained in the SOC of various benign, malignant, and tumors of undetermined nature (including cysts and polyps). CONCLUSIONS For ADEs that occur frequently with axitinib and are already contained in the drug instruction (e.g. hypertension, diarrhea), they should be adequately evaluated before administration, especially for patients with combined use of immune checkpoint inhibitors and patients with underlying hypertension; for ADEs with stronger signals and newer ADEs (e. g. death, disease progression, tumor progression), the patient’s disease progression should be closely monitored during the treatment period for potentially fatal ADEs; for its rare ADEs (e. g.immune-mediated nephritis, scrotal ulcer, non-infectious encephalitis), clinical validation should be further strengthened.