1.Tendinopathy and the extracellular matrix:to deeply study the key proteins and enzymes in the bone-tendon junction
Chunyu LIU ; Xiaoyan HAN ; Lin WANG
Chinese Journal of Tissue Engineering Research 2016;20(15):2265-2272
BACKGROUND:In recent years, research on tendinopathy is becoming a hot topic in the sports medicine and rehabilitation, and the extracelular matrix plays a considerable role in the occurrence, development and healing of tendinopathy.
OBJECTIVE: To explore the relationship between tendinopathy and the extracelular matrix.
METHODS: CNKI, VIP, CBM, Wanfang, PubMed and Embase were retrieved for articles related to colagen, protein, glycoprotein and matrix remodeling published since 2000.
RESULTS AND CONCLUSION:After tendinopathy, there are obvious changes in colagen, proteoglycans, glycoprotein and related enzymes; and these changes also affect the reconstruction and repair of tendinopathy. Currently, most of researches focus on the tendon rather than the bone-tendon junction. This means that there are no in-depth studies on the correlation between many important proteins and tendinopathy, and the effects of exercise intervention.
2.Study of the biological characteristics and antitumor activity of recombinant IFN-α-2B-BCG
Erlin SUN ; Chunyu LIU ; Ruifa HAN ; Xiaodong FAN
Chinese Journal of Microbiology and Immunology 2010;30(1):46-50
Objective To investigate the biological characteristics of recombinant hIFN-α-2B-BCG and its direct effect to bladder tumor cells in vitro. Methods BCG and recombinant BCG(rBCG) wild-type growth and morphology were compared. After 10 generations, hIFN-α-2B content was analyzed by ELISA, of bacteria in vivo and in vitro. The effects of rBCG on bladder cancer cells EJ, MB49, were detected by elec-tron microscopy and cell inhibitory rate from MTT. Results The normal BCG and rBCG had no significant difference in growth phase. They were both positive for acid-fast stain, maintaining the characteristics of cell's connection. While rBCG slightly larger than normal BCG, no else abnormality was obvious between them. IFN-α-2B was 997.2 pg/ml in secretions, 99.3 pg. In bacteria in vivo, 990.3 pg/ml in 10th genera-tion's sections, of rBCG. Compared with 1st generation, rBCG had no significant differences in morphology and interferon expression. Normal BCG and rBCG both had, anti-proliferation directly on bladder tumor cell in vitro, the rBCG is most effective in all. After rBCG cultivated with bladder tumor cell together, tumor cell's slow proliferation, detach, quantity decreasing and death were observed under microscope. Different degeneration in degree on most of tumor cells, disorganization on organelle, aggregation on chromatin, pyc-nosis on nucleolus, and cytoplasm lysis were on tumor cell under transmission electro microscopy. MTT as-say showed rBCG inhibited the proliferation of bladder caner cell and more active than normal BCG. Conclu-sion These results suggest that the rBCG have the same characteristics in growth phase as normal BCG, and stable properties in interferon expression and morphology by generations, rBCG has more anti-tumor effects in vitro than normal BCG.
3.Difluoromethylornithine suppresses growth and improves apoptosis of human leukemia Jurkat cells
Yushan REN ; Yu HAN ; Chunyu CAO ; Yanlin WANG
Journal of Third Military Medical University 1988;0(06):-
Objective To investigate whether difluromethylornithine (DFMO) can be used in the treatment of human leukemia. Methods The cell proliferation was detected by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)] assay after treatment of human lymphocyte Jurkat cells by DFMO (0 to 10 mmol/L) for 24 to 72 h. Enzyme activity of spermine oxidase (SMO) and acetylpolyamine oxidase (PAO) was determined by chemiluminesence assay. DNA fragmentation assay was used to evaluate cell apoptosis. Fluorescent dye assay was performed to determine the changes in mitochondrial membrane potential. Western blotting was used to determine Bax content. Casepase-3 enzyme activity was measured by spectrophotometric method. Results DFMO treatment inhibited the proliferation of Jurkat cells significantly in a dosage- and time-dependent manner (P
4.Polyamine analogue CPENSpm inhibits proliferation of human lung cancer cells by interfering polyamine metabolism
Yanlin WANG ; Yu HAN ; Taining YUAN ; Chunyu CAO ; Yongqin ZHOU
Chinese Pharmacological Bulletin 1987;0(01):-
Aim To study the effects of polyamine analogue CPENSpm on the human lung cancer line A549 in cell proliferation and apoptosis.Methods MTS was used to assay the cell proliferation,chemical analysis methods were used to determine the activities of enzymes in the polyamine metabolism,HPLC was performed to assay the intracellular concentration of polyamines,Sub-G1 and DNA fragmentation assays were used to determine the cell apoptosis.Results Treating A549 lung cancer cells by CPENSpm resulted in:①cell-growth inhibition and cell apoptosis;②inhibition of ODC(key enzyme in polyamine synthetic pathway)and activation of SSAT and SMO(key enzymes in polyamine catabolism);③great decrease of intracellular polyamine concentrations.MDL72527,the SMO inhibitor,can antagonize the effect of CPENSpm on inhibiting the proliferation of A549 cells.Conclusion CPENSpm inhibits proliferation and induces apoptosis of human A549 lung cancer cell line by interfering the polyamine metabolism,depleting intracellular polyamine contents that are need by quick-growth of cancer cells and inducing production of H2O2.
5.Effects of Guizhi Fuling Capsule on prostatic hyperplasia in experimental rats
Chunyu LIU ; Jianxin PAN ; Keping ZHANG ; Feng HAN ; Zhenlun GU ;
Chinese Traditional and Herbal Drugs 1994;0(09):-
Object To observe the effect of Guizhi Fuling Capsule (GFC) on prostatic hyperplasia in rats induced by testosterone propionate. Methods Seven days after the male rats were castrated, testosterone propionate (3 mg/kg) was sc injected in rats. Meanwhile, the rats were administered GFC (2.16, 1.08, 0.54 g/kg) once a day, and the treatment continued for 30 d. An hour after the final administration, the rats were put to death, and the weight and volume of prostates were measured. The construction of prostate cells was also observed under light microscope. Results In the GFC group, the weight and volume of prostate was significantly decreased compared with that in the model group (P
6.Prokaryotic expression and polyclonal antibody preparation of human spermine oxidase
Chunyu CAO ; Yu HAN ; Yushan REN ; Yanlin WANG
Journal of Cellular and Molecular Immunology 2009;25(10):920-923
AIM: To prepare recombinant human spermine oxidase (SMO) and polyclonal antibody against human SMO by gene recombination techniques. METHODS: Human SMO cDNA was amplified from total RNA of A549 cells through reverse transcription PCR. The cDNA was then cloned into pET-15b to construct SMO prokaryotic expression vector. After transforming, the vector was induced to express recombinant SMO by IPTG in E. coli BL21 (DE_3). Recombinant SMO was purified by Ni-NTA resin under denaturing condition and then was dialyzed to renature. The enzyme activity of recombinant SMO was analyzed by chemical fluorescent method. SMO polyclonal antibody was prepared by using recombinant human SMO protein purified by polyacrylamide gel electrophoresis as antigen to inoculate rabbit intradermally. The titer and specificity of anti-sera were determined by ELISA, Western blot and Immune Cell Chemistry. RESULTS: Purified and dialyzed recombinant human SMO has the specificicity of oxidizing the spermine. The polyclonal antibody has high titer and specificity against human SMO. CONCLUSION: This research established a method for prokaryotic expression, purification and polyclonal antibody preparation of human SMO. The method lays a foundation for the future functional research of SMO.
7.Comparative analysis of clinical data among 385 postoperative patients with thyroid benign and malignant nodules
Na KONG ; Xinyan CHEN ; Chunyu ZHANG ; Xue HAN
Chinese Journal of Endocrinology and Metabolism 2012;28(10):806-810
Objective To analyze the clinical features of 385 postoperative thyroid nodules patients,and to compare the correlation affected by different factors between benign and malignant thyroid nodules.Methods Total 385 patients underwent thyroid operation in general surgery department of our hospital were enrolled.Their clinical data,ultrasonic scanning of thyroid,and pathological results were compared between benign and malignant thyroid nodules.Results (1)264 cases with benign nodules (68.57%) and 121 cases with malignant nodules (31.43 %)were found among 385 cases.The average age of benign group was (48.1 ± 13.4) years old,while malignant nodules group was (45.6 ± 12.5) years old.(2) No gender difference was found between benign and malignant nodules group (P>0.05).(3) According to body mass index (BMI),the highest prevalence of malignant nodules was in overweight group,followed by obesity group.(4) Fasting blood glucose was (5.61 ± 1.07) mmol/L in benign group and (5.86±1.21) mmol/L in malignant group(P<0.05).(5) Thyroid-stimulating hormone (TSH) in benign nodules group was (1.62 ± 1.30) mIU/L,while in malignant nodules group,which was (2.02 ±2.59) mIU/L(P<0.05).(6) Irregular shape,unclear boundary,calcification,and lymph node metastasis were more frequent in malignant group than those in benign group under ultrasonic scanning (P<0.05).(7) Logistic regression analysis revealed that the existence of thyroid cancer was significantly associated with age,fasting blood glucose,ultrasonic images(lymph node metastasis,irregular shape,and calcification).Conclusion (1) Fasting blood glucose levels and TSH in the malignant nodules group are higher than those in the benign group.(2) A negative correlation exists between age and thyroid cancer prevalence.(3) Ultrasonic images,including lymph node metastasis,irregular nodule shape,and calcification,may suggest the high possibility of thyroid cancer.
8.Serial Processing of Exogenous and Endogenous Information Conflicts in Human Brain
Wei MAO ; Yuanyuan ZHANG ; Wei SUN ; Chunyu HAN ; Yuping WANG
Chinese Journal of Rehabilitation Theory and Practice 2009;15(8):728-731
Objective To investigate the cortical activities for processing exogenous stimulus-related information conflict and endogenous mental-related information conflict.Methods Event-related potentials (ERPs) were recorded while 15 healthy subjects performed a digit matching task. Each trial consisted of two sequentially presented digits (S1~S2), where S2 was either the same as S1 (S-) or different from S1 (S+) in digital value. Subjects pressed a button of a push-pad when S2 was the same as S1 and pressed the other button when S2 was different from S1 in the first session. Furtherly, they were required to calculate the difference between S1 and S2, and compare their calculation result with digit '3' in the second session. They pressed a button when the calculation result was equal to '3' (M-) and pressed the other button when it was not (M+). Three trial types were included: same numbers but their difference was not equal to '3' (S-M+), different numbers but their difference was '3' (S+M-) and different numbers but their difference was not '3' (S+M+). Results Following S2 onset, a component N270 was broadly elicited at all the scalps by S+, S+M- and S-M+. N270 and N400 were elicited in series by S+M+. The maximal amplitude of N270 was at the posterior scalp while the maximal amplitude of the N400 was at the central areas. Conclusion The different spatio-temporal distributions of the two negative potentials suggested that exogenous and endogenous information conflicts were serially processed in the human brain by the conflict processing system of multiple neural substrates.
9.The construction of phIFN-?-2B shuttle plasmid and its expression in BCG
Chunyu LIU ; Ruifa HAN ; Tengxiang MA ; Jiwu CHANG ; Yuzhi HAN ; Zhifang SUI
Chinese Journal of Urology 2006;0(S1):-
Objective To construct a new secreting recombinant hIFN-?-2B-BCG to provide a new tool for the treatment of bladder tumor. Methods BCG was genetically engineered to secrete recombinant human interferon-alpha 2B by transforming of shuttle plasmid phIFN-?-2B. Expression of hIFN-? was readily detectable by ELISA. Results The phIFN-?-2B was transformed in BCG correctly,and the value of hIFN-?-2B in supernatant of recombinant BCG culture was calculated to approximately 997.2 pg/ml. Conclusions This study demonstrates that the recombinant phIFN-?-2B can be expressed in BCG secretively. As the construction of the plasmid and its transformation and expression in BCG were accomplished successfully, a foundation of reformed BCG and new vaccine will be established.
10.Therapeutic Effects of Artemisia Argyi Ferment Substance on Systemic Candida Albicans Infection
Jing BAI ; Lei HU ; Li ZHANG ; Chunyu TIAN ; Dequan PANG ; Haimei BO ; Shuying HAN
Herald of Medicine 2014;(11):1438-1441
Objective To investigate the therapeutic effects and mechanism of Artemisia argyi ferment substance on systemic Candida albicans infection. Methods The model of systemic Candida albicans infection was established in immunosuppressed mice. The model mice were randomly divided into the model control,Artemisia argyi ferment substance( AAFS) at different doses(100,200,and 400 mg·kg-1 )and fluconazole group(20 mg·kg-1 ),30 mice in each. Mice in each treatment group were given therapeutic drugs by gavage for 5 consecutive days,twice daily. The survival of mice was determined 21 days after the model was set up. The serum levels of IFN-γand IL-2 were determined by ELISA. The proliferation activity of T lymphocyte in the spleen was detected by MTT assay. The number of living fungi in liver and kidney tissues was counted. Results Compared with the model control,AAFS at middle and high doses and fluconazole significantly increased the survival rate of mice,the serum levels of IFN-γand IL-2,and the proliferation activity of T lymphocyte in the spleen,but decreased the number of living fungi in tissues(P〈0. 01). Compared with low dose AAFS,middle and high doses of AAFS and fluconazole showed significantly different effect on each index(P〈0. 05 or P〈0. 01),but there was no difference among these groups(P〉0. 05). Conclusion AAFS at 200-400 mg·kg-1 has inhibitory effects on systemic Candida albicans infection in mice,the mechanism of which is related to increasing the proliferation of T lymphocyte in spleen and the levels of IFN-γand IL-2 in serum.