1.ATRA combined with IFN-α2a promotes iodine uptake of FTC-133 follicular thyroid carcinoma cells
Zhen ZHAO ; Guohua SHEN ; Huawei CAI
Journal of Medical Postgraduates 2016;29(9):910-912
Objective Some drugs can effectively improve the uptake of 131 I in dedifferentiated thyroid cancer .This study was to investigate the effect of all-trans retinoic acid (ATRA) combined with interferon-α2a (IFN-α2a) on the iodine uptake and re-tention rate of FTC-133 follicular thyroid carcinoma cells . Methods FTC-133 cells were cultured in the presence of 2 μmol/L AT-RA and/or 1500 IU/mL IFN-α2a for 72 hours.The the cells were collected for measurement of the uptake and retention rate of Na 125 I. Results The Na125 I uptake of the FTC-133 cells was significantly increased after 72-hour incubation with 2μmol/L ATRA and 1500 IU/mL IFN-α2a (2423.1 ±237.5) as compared with those of the control (1279.5 ±102.8), ATRA (1438.2 ±149.6), and IFN-α2a groups (1355.3 ±198.4) (P<0.05).Statistically significant differences were observed in the retention rate of Na 125I in the FTC-133 cells at different time points in the each of the blank control , 2μmol/L ATRA, 1500 IU/mL IFN-α2a, and combination groups (P<0.01), but not in that of Na125I among these groups (P<0.05). Conclusion ATRA in combination with IFN-α2a can promote the Na125 I uptake but cannot improve the Na 125 I retention rate in FTC-133 cells.
2.Research advancement of pro-apoptotic mechanisms of GRIM-19 and its related proteins
Guohua SHEN ; Huawei CAI ; Zhen ZHAO
Journal of Medical Postgraduates 2015;(3):303-307
Cancer development is a complex process that involves multiple genetic changes and multiple signaling pathways . Recent findings show that the GRIM-19 is a novel apoptosis regulation gene , and its gene mutations and loss of protein expression have been observed in many tumor types such as urinarysystem tumor , digestive system neoplasm , which are closely related to cancer devel-opment.Thus, GRIM-19 may be a potential target for gene therapy .Pro-apoptotic mechanisms of GRIM-19 and its related proteins such as STAT3,GW112,p16INK4aare overviewed in this article.
3.Analysis of pedicle screw misplacement in the thoracolumbar spine of 19 cases
Yafeng ZHANG ; Jianwei WANG ; Jianping CAI ; Xiaowu TIAN ; Huawei ZOU
Chinese Journal of Tissue Engineering Research 2009;13(39):7773-7776
OBJECTIVE: To analyze the causes for misplacement of pedicle screw in thoracolumbar spine.METHODS: From January 2002 to January 2008, 19 patients with misplacement in thoracolumbar spine were treated in Department of Orthopedics, Wuxi Affiliated Hospital of Nanjing University of Chinese Medicine, including 12 males and 7 females with an average age of 52.5 years (range 23-68 years). The diagnoses were thoracolumbar fracture in 5 cases, lumbar spondylolisthesis in 8 and degenerative lumbar disease in 6. The fixation systems were Steffee used in 4 cases, DRFS in 3, RF in 6, AF in 4 and GSS in 2. X-ray and CT scanning were used to observe pedicle screw location, including screw,pedicle and membranous sac and great vessels.RESULTS: The time of misplacement of pedicle screw was 5-69 days with an average time of 18.5 days, including 7 cases of screw penetrating into lateral cortex, 4 of screw penetrating into medial cortex, 2 of screw penetrating into pedicle cortex, 2 of overplacement, 2 of intervertebral foremen placement and 2 of intervertebral space placement.CONCLUSION: The causes for screw misplacement were anatomic variation and poor surgical skills, and the key factors in precise insertion of pedicle screw are fine surgical skills, carefully study of preoperative image and the intra-operative monitoring.
4.Metabonomics and its application in thyroid diseases
Zhen ZHAO ; Yu MA ; Guohua SHEN ; Huawei CAI
Chinese Journal of Nuclear Medicine and Molecular Imaging 2017;37(1):49-52
As an important branch of systematic biology, metabonomics is an emerging discipline following genomics, proteomics and transcriptomics. It focuses on the quality and quantity changes of the small molecular metabolites in appointed physiological or pathological state, and provides important informa?tion for studies of diseases mechanism, clinical early diagnosis and prognosis prediction. Compared with tra?ditional clinical diagnostic method ( single index) , metabonomics method can more comprehensively analyze metabolites changes in the body. This review introduces the metabonomics and summarizes its application in thyroid diseases.
5.advanced aged patients with Bipolar artificial femoral head replacement in the treatment of advanced aged ;patients with femoral neck fracture
Huawei CHEN ; Yanlu CAI ; Wanan CHEN ; Ruizhong LIN
Chinese Journal of Primary Medicine and Pharmacy 2013;20(z2):18-19
Objective To explore the clinical curative effect of the bipolar artificial femoral head replace-ment in the treatment of advanced aged patients with femoral neck fracture .Methods 32 advanced aged patients with femoral neck fracture were treated with the bipolar artificial femoral head replacement ,the effect was observed . Results 32 patients were stage Ⅰincision healing ,and after two weeks all could ambulate .Followed up for 6 to 36 months,two cases of dislocation ,two cases of postoperative pain ,3 cases of deep venous thrombosis ,one case of hip limited functionality.Harris score function assessment:excellent 24 cases(74%),good in 5 cases(16%),3 cases (10%).Conclusion Bipolar artificial femoral head replacement is the effective treatment of advanced aged patients with femoral neck fracture,with shorter operation time,small trauma,early and activities,and reduce the complications and reduce mortality rate etc .
6.Effect of 5-Fluorouracil and Radiotherapy on the Apoptosis of Human Cervical Adenocarcinoma HeLa Cells
Bin YANG ; Min CAI ; Hua YAN ; Huawei ZHOU ; Fan YANG ; Yuan LIU
Journal of China Medical University 2010;(1):24-27
Objective To investigate the effect of 5-fluorouracil (5-FU) combined with radiotherapy (RT) on the apoptosis of human cervical cancer HeLa cells.Methods The HeLa cells were divided into four groups treated with chemotherapy(ChT),RT,RT+ChT, and control groups.The non-cytotoxic concentration (48 h IC_(50)) of HeLa cells treated with 5-FU was determined by methyl thiazolyl tetrazolium (MTT) and the apoptosis rates of HeLa cells detected by flow cytometry with annexin V-FTTC and PI double labeling.The morphologic changes of the apoptosis cells were observed under fluorescence microsope.Results ChT,RT and RT+ChT treatment could induce the apoptosis of HeLa cells with the strong induction of the combined treatment (P<0.05).Conclusion 5-FU combined with RT can obviously enhance the apoptosis of HeLa cells, which provides a reliable experimental evidence for clinical use of 5-FU combined with RT in the intervention of human cervical cancer.
7.Cloning and recombinant expression of human soluble TRAIL in Pichia pastoris.
Yingying LI ; Lin WAN ; Hao YANG ; Shan LIU ; Huawei CAI ; Xiaofeng LU
Journal of Biomedical Engineering 2010;27(6):1307-1326
TNF-related apoptosis-inducing ligand (TRAIL) is a member of factor TNF family, which could be potentially developed as novel antitumor agent due to its selective and efficient induction of apoptosis in tumor cells. Gene recombinant expression is an important tool for production of pharmaceutical protein. In this paper, the gene encoding human soluble TRAIL (114-281aa fragment) was cloned by PCR and then inserted into the Pichia Pastoris expression vector pPIC9K. The transformants were double-screened on plates containing neomycin G418 and many clones with high levels of G418-resistance were selected for further studies on protein expression. The recombinant human soluble TRAIL was secreted into the BMMY media under the condition of 3% methanol. And the recombinant protein was purified to homogeneity (-80% purity) by using Ni-agarose affinity chromatography. The yield of this protein is about 1-2 mg per liter culture. Cell viability assays demonstrated that human soluble TRAIL was cytotoxic in both leukemia cells Jurkat and lung cancer cells A549. After treatment with 0.05 microg/ml TRAIL, the survival rate of Jurkat cells was about 10%. The expressed TRAIL showed dose-dependent cytotoxicity in A549 cells within the range of 0.1-1 microg/ml. When the protein concentration reached 1 microg/ml, the survival rates of A549 cells were about 30%. However, the recombinant human soluble TRAIL did not show obvious cytotoxicity in human skin fibroblast cells (HSF) at concentrations tested. There results demonstrate that human soluble TRAIL is selectively cytotoxic in tumor cells. The expression system constructed in this experiment might contribute to further production of soluble TRAIL and TRAIL-based novel fusion proteins in large quantities.
Antineoplastic Agents
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pharmacology
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Cell Line, Tumor
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Cloning, Molecular
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Cytotoxicity Tests, Immunologic
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methods
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Genetic Vectors
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genetics
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Humans
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Pichia
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genetics
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metabolism
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Recombinant Proteins
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biosynthesis
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genetics
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pharmacology
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TNF-Related Apoptosis-Inducing Ligand
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biosynthesis
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genetics
8.Expression and purification of recombinant human cytochrome C in Escherichia coli.
Huawei CAI ; Lin WAN ; Hao YANG ; Lihong CHEN ; Xiaofeng LU
Journal of Biomedical Engineering 2007;24(3):620-625
Cytochrome C plays important roles in electron transferring, oxidative stress and apoptosis. In this study, soluble cytochrome C was accumulated in cytoplasm of E. coli by utilizing the co-expression of human cytochrome c and yeast heme lyase from a single plasmid. After ultrasonic disruption of the bacteria, a lot of contaminated proteins were discarded by addition of 350 g/L ammonium sulfate into the supernatant. Then the recombinant human cytochrome C was purified to 80% homogeneity after two times cation exchange chromatography on SP-Sepharose Fast Flow. Yields of cytochrome C greater than 10 mg per liter culture were attained. This efficient system for producing human cytochrome C is helpful for us to understand the roles of this protein in biological processes and therapy of human diseases relevant to apoptosis and oxidative stress.
Cytochromes c
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biosynthesis
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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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Genetic Vectors
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Humans
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Recombinant Proteins
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biosynthesis
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genetics
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isolation & purification
9.Enhancement of cytotoxicity of cantionic antimicrobial peptide in tumor cells by conjugation to cell-penetrating peptide.
Shan LIU ; Hao YANG ; Huawei CAI ; Lin WAN ; Xiaofeng LU
Journal of Biomedical Engineering 2011;28(1):110-114
Due to their lower risk for induction of resistance, membrane-active antimicrobial peptides with anticancer effect are attractive in cancer therapy. Because cell binding contributes to the cytotoxicity of peptide, it is possible to enhance the cytotoxicity of antimicrobial peptide in tumor cells by conjugation to a cell-penetrating peptide (CPP). In this paper, a fusion peptide MPGA by conjugation of antimicrobial peptide MP to CPP Antp at its N-terminus was constructed. After compared the cytotoxicity of unconjugated MP with that of the fusion peptide, it was found that MPGA showed higher cytotoxicity than that of unconjugated MP. And the fusion peptide MPGA induced cell death in tumor cells by membrane disruption. These results demonstrated that the cytotoxicity of antimicrobial peptide can be significantly enhanced by conjugation to CPP, which might be an effective way to develop novel anticancer drugs.
Antimicrobial Cationic Peptides
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pharmacokinetics
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pharmacology
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Antineoplastic Agents
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pharmacokinetics
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pharmacology
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Cell Line, Tumor
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Cell Membrane Permeability
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drug effects
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Humans
10.Expression and prognostic significance of nuclear matrix protein 4 in hepatocellular carcinoma
Jinhai LI ; Huawei ZHAI ; Guangzheng SUN ; Haifeng ZHANG ; Minghui ZHU ; Yu CAI ; Shenghua PAN ; Shuqun LI
Chinese Journal of Hepatobiliary Surgery 2024;30(2):87-92
Objective:To investigate the expression of nuclear matrix protein 4 (NMP4) in hepatocellular carcinoma (HCC), and its relationship with clinicopathological features and survival prognosis of patients.Methods:The clinical data of 100 HCC patients who were treated with radical resection of liver cancer in the Department of Hepatobiliary Surgery of the Third Affiliated Hospital of Wenzhou Medical University from July 1, 2014 to July 1, 2019 were retrospectively analyzed. There were 63 males and 37 females, aged (58.5±10.4) years old. Immunohistochemical method was used to detect the expression of NMP4 protein in HCC cancer tissue and the corresponding adjacent normal tissue. According to the expression of NMP4 in HCC tissues, 100 patients were divided into two groups: the NMP4-positive expression group ( n=62) and the NMP4-negative expression group ( n=32). Univariate analysis was performed on the relationship between NMP4 expression and clinical pathological features as well as overall survival of HCC patients. Cox multivariate analysis was performed on the factors influencing postoperative prognosis of HCC patients. Results:Immunohistochemistry results showed that NMP4 was primarily expressed in the nucleus, the positive expression rate of NMP4 in HCC tissues was higher than that in adjacent non-cancerous tissues [62.0% (62/100) vs. 8.0%(8/100)], and the difference was statistically significant ( χ2=2.12, P=0.003). Univariate analysis revealed that the overall survival of HCC patients was correlated with the degree of tumor differentiation, tumor length, BCLC stage, number of tumor foci, vascular tumor thrombus and expression of NMP4 (all P<0.05). Cox multivariate analysis revealed that low differentiation, high BCLC stage (stage C), number of tumor foci (≥3), and positive expression of NMP4 were independent risk factors affecting postoperative survival and recurrence-free survival of HCC patients. The median overall survival and median recurrence-free survival of HCC patients in the NMP4-positive expression group were 22.3 months and 11.5 months, respectively. In contrast, that in the NMP4-negative expression group were 40.6 months and 19.4 months, respectively. The cumulative survival rate and recurrence-free survival rate of HCC patients in the NMP4-positive expression group were lower than those in the NMP4-negative expression group, and the differences were statistically significant (both P<0.05). Conclusion:Positive NMP4 expression was closely correlated with malignant biological progression and poor prognosis of HCC patients.