1.Cytokine-induced killer cells in cancer treatment
Xuefeng HANG ; Zhenyu DING ; Xiaodong XIE
Journal of International Oncology 2012;39(5):344-347
Cytokine-induced killer cells (CIK) is the fourth largest cancer treatment after surgery,chemotherapy and radiotherapy,and it is the development direction of cancer treatment.It is a new type of immune cell,and it is named after natural killer cell samples T lymphocytes as it express CD3 and CD56.Currently,CIK treatment has a broader range of clinical applications,and it has achieved the better clinical efficacy in the blood system cancer and solid tumors,The CIK adoptive immunotherapy is considered to be a new hope for the anticancer treatment.
2.Effect of active compound components of SQ on MMP-1,TIMP-1,VEGF,t-PA and PAI-1 of in? ammatory endometrial cell
Qing DING ; Zhaoling YOU ; Zhenyu TAN
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(03):-
Objective :To study the effect of MMP-1,TIMP-1,VEGF and t-PA,PAI-1 in the inflammatory process of endometrium cell and the active components of SQ.Methods: ICC,ISH and WB was used to test the level of protein of MMP-1,TIMP-1,VEGF and t-PA,PAI-1 and mRNA of MMP-1,TIMP-1 and VEGF of endometrium cell in regular group,pattern group,GongXueNing group and SQ group.Result: In contrast with model group,the level of MMP-1protein and mRNA expression obviously decreased,but the level of TIMP-1 and VEGF increased in SQ group,there were signifi cant differences(P
3.Expression of signal transducer and activator of transcription 3 protein in development of embryo liver in mice
Lu MA ; Zegui LI ; Hua JI ; Zhenyu DING ; Yizhan XING
Chinese Journal of Tissue Engineering Research 2005;9(7):188-190
BACKGROUND: Signal transducer and activator of transcription 3 (STAT3) protein is widely expressed in cells and tissues of different type and. It is the important component of signal transduction and transcription chain that participates in the regulation of various physiological functions of cells like proliferation and differentiation. However, there is no literature reported regarding if there is STAT3 expression and whether it participates in embryo liver development both at home and abroad.OBJECTIVE: To explore the effects and mechanism of STAT3 signal transduction route on development of embryo liver by. Studying the expression of STAT3 protein during embryo liver development in mice.DESIGN: Randomized controlled study based on STAT3 protein.SETTING: Department of histology and embryology in a military medical university of Chinese PLA.MATERIALS: The study was completed in the Department of Histology and Embryology, Third Military Medical University of Chinese PLA during January to June 2004. Kunming mice were provided by Animal Centre of Third Military Medical University of Chinese PLA. Ten male mice and 20 female mice with body mass during 25 to 30 grams were randomly chosen.METHODS: Serial frozen sections of mouse embryo was performed in the 13.5 days ( n = 8), 14.5 days ( n = 9) and 15.5 days ( n = 7) after mating respectively. One of the every 2 slices was used as negative control by replacing first antibody of STAT3 with 0. 01 mol/L PBS. Immunochemical technique and immunofluorescence method were used to display the expression of STAT3 protein in embryo liver development in mice.MAIN OUTCOME MEASURES: Expression of STAT3 protein in serial frozen sections of mouse embryo.RESULTS: There was strong expression of STAT3 protein in liver cells in E13.5 days mouse embryo while the expression was decreased in E14.5 days and E15.5 days mouse embryos.CONCLUSION: STAT3 protein has participated in the development of embryo liver. The expression of STAT3 in liver development can be used to explain the mechanism of liver regeneration and provide experimental data for organ repair.
4.Effect of genistein on cell cycle and apoptosis in PC12 cells transfected with APP695MT gene
Fengbin WANG ; Zhenyu DING ; Youhua KONG ; Yanan WANG ; Ning LI
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(2):123-125
ObjectiveTo investigate the effect of Genistein (GST) on cell cycle and apoptosis in PC12 cells transfected App695MT gene.MethodsPC12 cells were transfected with pIRES2-EGFP plasmid or pIRES2-EGFP/APP695MT expression plasmid,and then were divided into control vectortransfected group,APP695 transfected group and GST treatment group.Flow cytometry was applied to detect cell cycle and apoptosis,laser confocal microscope was used to observe morphological changes of cell apoptosis.ResultsCompared with control vectortransfected group,PC12 cells in APP695 transfected group increased significantly in G0 and G1 phase,and less into S phase,cell proliferation index was decreased significantly( (55.6 ±0.57)%,P<0.0l ),apoptosis rate was increased significantly( (77.10 ± 12.53)%,P<0.01 ).Emitted red fluorescence increased significantly when cell death,cell body swelling,organelle disintegration,nuclear condensation or fragmentation.Compared with APP695 transfected group,PC12 cells in GST( 15μ mol/L) treatment group,decreased significantly in G0 and G1 phase,and more into S phase,cell proliferation index was increased significantly ( ( 61.57 ± 0.47 ) %,P < 0.01 ),apoptosis rate was decreased significantly ( (46.00 ± 8.43 ) %,P < 0.01 ).Cell death was significantly reduced red fluorescence,emitted green fluorescence was significantly enhanced,compared with APP695 transfected group cell morphology improved.ConclusionGST can improve APP695MT gene caused cell cycle arrest,promote cell to S phase transition,reduce apoptosis rate in PC12 cells,and have a protective effect on transfected cells.
5.Effect of genistein on the expression of Bax and Bcl-2 protein in PC12 cells transfected with mutant APP695 type
Zhenyu DING ; Cuixiang ZHAO ; Fengbin WANG ; Bing LIU ; Chunyan TAN ; Yanan WANG
Chinese Journal of Behavioral Medicine and Brain Science 2013;22(9):791-793
Objective To investigate the effect of genistein on the expression of Bax and Bcl-2 protein in PC12 cells transfected with mutant APP695 Type.Methods PC12 cells were transfected with pIRES2-EGFP plasmid or pIRES2-EGFP/APP695MT expression plasmid,and then divided into normal control group,Unladen group,APP695 transfected group,GST treatment group(5 μmol/L,15 μmol/L).The Bax and Bcl-2 protein expression in PC12 cells was measured by Immunocytochemistry SABC and Western-blot.Results Immunocyto-chemistry SABC showed that:compared with the normal control group,the expression of bax protein in APP695 transfected group was strongly increased((78.02 ± 1.26) %,P<0.01)) and the expression of bcl-2 protein was very weak oppositely ((15.40 ± 0.72) %,P<0.01)).Compared with APP695 transfected group,the expression of bax protein in cells of 15umol/L GST treatment group was significantly decreased((22.35 ± 0.49) %,P<0.01))and the expression of bcl-2 protein in these cells was strongly increased ((68.11 ± 0.24) %,P <0.01)).Western-Blot showed that:compared with the normal control group,the expression of bax protein in APP695 transfected group was markedly increased and the expression of Bcl-2 protein was significantly decreased.Compared with the APP695 transfected group,the expression of Bax protein was markedly decreased and the expression of Bcl-2 protein was significantly increased in the cells of the GST treatment groups.Conclusion GST can reduce the expression of bax protein and the increased expression of Bcl-2 protein has protective effect on PC12 cells transfected with APP696MT.
6.Influence of MTDH gene down regulation on proliferation and apoptosis of human breast cancer SK-BR 3 cells
Cheng DU ; Zhaozhe LIU ; Zhenyu DING ; Fang GUO ; Dongchu MA ; Xiaodong XIE
Journal of Endocrine Surgery 2013;7(5):359-363
Objective To investigate if down regulation of MTDH could inhibit proliferation and induce apoptosis in breast cancer SK-BR-3 cells.Methods RNA interference was employed to reduce MTDH expression in human breast cancer SK-BR-3 cells.Western blot assay was applied to measure the down regulation of MTDH.MTT assay was performed to assess the proliferation of SK-BR-3 cell.Flow cytometry was employed to detect cell cycle and apoptosis.Western blot assay was applied to detect the molecular alterations that was associated with cell proliferation,cell cycle and apoptosis.Results MTDH down regulation significantly inhibited cell proliferation.48 hours and 72 hours after trasnfection,the absorbance value(A value)in blank control,negative control and treatment group was (2.0 ± 0.1) vs (1.9 ± 0.3) vs (0.9 ± 0.1) (P =0.02) and (2.7 ± 0.2) vs (2.5 ± 0.4) vs (1.3 ± 0.2) (P =0.008).MTDH down regulation resulted in accumulation of the G0/G1 phase cells and reduction of S and G2/M phase cells.Moreover,MTDH down regulation significantly induced cell apoptosis.The cell apoptosis rate in blank control,negative control and trial group was (1.3 ± 0.2) %,(1.4 ± 0.3) % and (19.6 ± 2.7) % (P =0.012).MTDH down regulation resulted in a decreased expression of cyclinD1 and Bcl-2,an increased expression of P21 and the activation of caspase-3.Conclusions Reduced MTDH expression in SKBR-3 cells can inhibit proliferation and induce apoptosis,which may be associated with decreased expression of cyclinD1 and Bcl-2,an increased expression of P21 and the activation of caspase-3.
7.Motor capacity early after cardiac surgery
Shijie LU ; Zhenyu LI ; Zhiyu QIAO ; Yaodong DING ; Yi YANG ; Shichao GUO ; Yu XIA ; Yipeng GE ; Junming ZHU ; Tie ZHENG
Chinese Journal of Physical Medicine and Rehabilitation 2021;43(3):231-235
Objective:To observe the motor capacity of patients early after cardiac surgery using a cardiopulmonary exercise test.Methods:Patients who had performed a cardiopulmonary exercise test within 3 months after cardiac surgery were included in this retrospective study. Patients who took the test within 30 days of the operation formed a discharge group ( n=20), those within 30 to 60 days and 60 to 90 days formed the one month and two month groups ( n=10 for both). The discharge group was further divided into an aortic surgery group ( n=9), a bypass surgery group ( n=6) and a valve surgery group ( n=5) according to their procedure. The exercise capacity of each person was measured in terms of the changes in heart rate and systolic pressure from the resting to the anaerobic threshold stage. Anaerobic threshold, peak oxygen uptake and carbon dioxide ventilation equivalent were also recorded. Results:All of the patients completed the cardiopulmonary exercise test above the anaerobic threshold, and no adverse events such as exercise accidents occurred. At the anaerobic threshold the average heart rate of the discharge group was (8.8±7.1)bpm, significantly lower than the averages of the one month and two months groups: (17.0±5.9) and (18.3±10.5)bpm respectively. The average anaerobic thresholds and peak oxygen uptakes of the 1 month and 2 months groups were not significantly different, but they were all significantly higher than the discharge group′s averages. There were, however, no significant differences among the groups in the average changes in their systolic pressure and carbon dioxide ventilation equivalent. Moreover, the average anaerobic threshold and peak oxygen uptake of the aortic surgery group and the bypass surgery group were significantly lower than the valve surgery group′s averages.Conclusions:Postoperative motor ability after cardiac surgery improves significantly for at least 30 days. Patients who have received aortic or bypass surgery have significantly lower exercise capacity than those after valve surgery.
8.High titer ethanol production from an atmospheric glycerol autocatalytic organosolv pretreated wheat straw.
Liang WANG ; Jianquan LIU ; Zhe ZHANG ; Feiyang ZHANG ; Junli REN ; Fubao SUN ; Zhenyu ZHANG ; Cancan DING ; Qiaowen LIN
Chinese Journal of Biotechnology 2015;31(10):1468-1483
The expensive production of bioethanol is because it has not yet reached the 'THREE-HIGH' (High-titer, high-conversion and high-productivity) technical levels of starchy ethanol production. To cope with it, it is necessary to implement a high-gravity mash bioethanol production (HMBP), in which sugar hydrolysates are thick and fermentation-inhibitive compounds are negligible. In this work, HMBP from an atmospheric glycerol autocatalytic organosolv pretreated wheat straw was carried out with different fermentation strategies. Under an optimized condition (15% substrate concentration, 10 g/L (NH4)2SO4, 30 FPU/g dry matter, 10% (V/V) inoculum ratio), HMBP was at 31.2 g/L with a shaking simultaneous saccharification and fermentation (SSF) at 37 degrees C for 72 h, and achieved with a conversion of 73% and a productivity of 0.43 g/(L x h). Further by a semi-SFF with pre-hydrolysis time of 24 h, HMBP reached 33.7 g/L, the conversion and productivity of which was 79% and 0.47 g/(L x h), respectively. During the SSF and semi-SSF, more than 90% of the cellulose in both substrates were hydrolyzed into fermentable sugars. Finally, a fed-batch semi-SFF was developed with an initial substrate concentration of 15%, in which dried substrate (= the weight of the initial substrate) was divided into three portions and added into the conical flask once each 8 h during the first 24 h. HMBP achieved at 51.2 g/L for 72 h with a high productivity of 0.71 g/(L x h) while a low cellulose conversion of 62%. Interestingly, the fermentation inhibitive compound was mainly acetic acid, less than 3.0 g/L, and there were no other inhibitors detected, commonly furfural and hydroxymethyl furfural existing in the slurry. The data indicate that the lignocellulosic substrate subjected to the atmospheric glycerol autocatalytic organosolv pretreatment is very applicable for HMBP. The fed-batch semi-SFF is effective and desirable to realize an HMBP.
Biofuels
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Carbohydrates
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chemistry
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Cellulose
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chemistry
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Ethanol
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metabolism
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Fermentation
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Furaldehyde
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chemistry
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Glycerol
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chemistry
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Hydrolysis
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Triticum
9.Association between activity of nuclear factor-kappa B and angiotensin system in renal tissues of diabetic rats
Helin DING ; Ying GUO ; Mingtong XU ; Shaoling ZHANG ; Lihong CHEN ; Feng LI ; Zhenyu ZHU ; Yiqun DENG ; Zuzhi FU
Chinese Journal of Tissue Engineering Research 2006;10(20):184-186
BACKGROUND: Nowadays, angiotensin Ⅱ plays an important role in onset of diabetic nephropathy. Therefore, the nuclear factor-κB may have adjustive effects on angiotonin system of kidney tissue of diabetic rats. OBJECTIVE: To observe the relationship of activity of inhibitive nuclear factor-κB with angiotensin Ⅱ and its type 1 receptor mRNA expression of renal tissue of diabetic rats. DESIGN: Completely randomized group design, control experiment. MATERIALS: The experiment was conducted at the Experimental Animal Center, Sun Yat-sen University of Medical Sciences between March and April 2000. Fifty-one pure breed clean grade male Wistar rats were select ed. METHODS: ①Models were established in 39 rats. Streptozotocin dissolv ing in citric acid buffer (0.1 mmol/L,pH=4.5) were given to establish dia betic models with 60 mg/kg intraperitoneal injection. If the fasting blood glucose maintained above 13.9 mmol/L, the establishment of models was successful. The thirty-nine rats were randomly assigned into 3 groups: model group (n=17, without other interventional measure, feeding normally) and pyrrolidine dithiocar2. Bamate (PDTC) (active inhibitor of nuclear fac tor-κB) interventional group [n=22, PDTC at the dose of 20 mg/kg were given with intraperitoneal injection, twice a day]. Other 12 rats were as normal control group, did not make into diabetic models with normal breeding. ②After feeding for 18 weeks kidneys were got in every group. The activity of nuclear factor-κB was detected with electrophoretic mobility shift assay. The expression of type 1 receptor mRNA of angiotensin Ⅱ was measured with reverse transcription polymerase chain reaction (RT-PCR). Contents of angiotonin Ⅰ and angiotensin Ⅱ were tested with Radio Im munoassay (RIA). Activity of rennin was referred to that the result of the level of angiotonin Ⅰ at 37 ℃ water bath subduced to that at 4 ℃. ③Dif ference of measurement data was compared with single factor analysis of variance. After normal transformation, the non-normal distribution data were conducted with statistical disposal. MAIN OUTCOME MEASURES: Comparison of contents of angiotensin Ⅰ and Ⅱ, activities of rennin and nuclear factor-κB and expression of type 1 receptor mRNA of angiotensin Ⅱ in renal tissues of rats of each group. RESULTS: In the normal control group, model group and PDTC interven tional group 1, 6 and 13 rats were dropped out, respectively, so 11, 11 and 9 rats in each group were involved in the result analysis. ①Activity of nu clear factor-κB: It was higher significantly in the model group than that in the normal control group and PDTC interventional group (P < 0.01 ). It was similar between the normal control group and the PDTC interventional group. ②Activity of rennin of renal tissue: It was similar among the 3 groups. ③Content of angiotonin Ⅰ of renal tissue: It was higher obviously in the model group that that in the normal control group and the PDTC interventional group (P < 0.01 ). ④Content of angiotensin Ⅱ in renal tissue: It was similar between the model group and the normal control group. It was lower markedly in the PDTC interventional group than that in the model group and the normal control group (P < 0.01 ). Expression of type 1 receptor mRNA of angiotensin Ⅱ: It was lower remarkably in the model group than that in the normal control group (P < 0.01 ). It was lower dis tinctly in the PDTC interventional group than that in the model group and the normal control group (P < 0.01 ). CONCLUSION: The increase of activity of nuclear factor-κB in renal tissue of diabetic rats can inhibit the activity of nuclear factor-κB, which will induce the reduction of the level of angiotensin Ⅱ and expression of type 1 receptor mRNA of angiotensin Ⅱ in renal tissue of diabetic rats.
10.Experimental study on the compound culture of small intestinal submucosa with synovial mesenchymal stem cells
Song CHEN ; Song PENG ; Peiliang FU ; Yuli WU ; Zheru DING ; Qi ZHOU ; Ruijun CONG ; Haishan WU ; Zhenyu XU
Chinese Journal of Orthopaedics 2014;(10):1059-1067
Objective To investigate the biosecurity and biocompatibility of small intestinal submucosa (SIS) as scaffold for tissue engineering and to explore the possibility to induce synovial mesenchymal stem cells (SMSCs) differentiate into cartilage with SIS as the scaffold and SMSCs as the seed cells. Methods The SMSCs were isolated and cultured from the synovial mem-brane of knee joints of rabbits by a conventional method. The SIS was harvested from pigs according to the physical method and Abraham's method. 4 rabbits are divided into the experimental group and control group. The biosecurity of SIS as scaf-folds were investigated in pyrogen test, skin sensitization test and systemic toxicity test. The SMSCs and SIS were co-cultured in vitro and induced to differentiate into cartilage to explore the biocompatibility of SMSCs and SIS, the proliferation and differ-entiation ability of SMSCs on the scaffold of SIS. The varietyies were identified by the microscope. Results The SIS isolated with the physical method and Abraham's method is a milky and translucent membrane with a smooth surface. Under the electron microscope, SIS presented a porous Stereoscopic three-dimensional network structure, which is formed by fibrous tissues' intertex-ture. Its' porosity was about 80%and its aperture was 100-200μm. Meanwhile, the biosecurity and biocompatibility of SIS were also fine. In the trial that the SMSCs and SIS were co-cultured in vitro, the SMSCs can grow, adhere to and differentiate on the sur-face of SIS and into the hollows very well. It can also secrete extracellular matrix. Through scanning microscope observation, cells contact with each other by their neuritis, or adhered to the wall of hole by cellular protrution. On the surface of SIS, the SMSCs maintain the property that it can easily differentiate into the chondrogenic lineage in the chondrogenic medium. Immunochemistry staining showed positive expression of type II collagen post 21 days of co-cultrue. Conclusion SIS can be used as the scaffold to construct tissue engineering meniscus as it has good biosecurity and biocompatibility with SMSCs without disturbing the cell form or inhibiting the growth and function of SMSCs.