1.The effects of protein phosphatase 2A inhibitors on the viability of pancreatic cancer cell and its mechanism
Wei LI ; Zheng CHEN ; Feiran GONG ; Yang ZONG ; Yi MIAO ; Min TAO ; Zekuan XU
Chinese Journal of Digestion 2012;32(1):42-45
ObjectiveTo investigate the effects of protein phosphatase 2A (PP2A) inhibitors on the viability of pancreatic cancer cell line PANC-1 and its mechanism.MethodsPANC-1 cells were treated with PP2A inhibitors Cantharidin or Okadiac acid.The activity degree of NF-κB pathway was tested by Western blot.NF-κB pathway was blocked from all sectors by PP2Acα plamid transfection,NF-κB inhibition of protein kinase α (IKKα) and NF-κB inhibitor α (IκBα) dominant negative mutant and p65 interfering plasmid.Cell viability was determined by MTT.ResultsPP2A inhibitors could induce phosphorylation of IKKα,further phosphorylation of IκBα and degradation and followed by the release of p65 into nucleus.When PP2Acα,IKKα dominant negative mutant and IκBα dominant negative mutant were overexpressed,or p65 was interfered,the inhibition rate of Cantharidin on cell viability decreased (31.85±13.37) %,(23.48±8.98)%,(22.63±5.81)% and (20.88±3.24)%respectively,and the inhibition rate of Okadiac acid on cell viability decreased (40.17 ± 11.65)%,(27.34±14.28)%,(24.85±3.39)% and (27.08±3.81)% respectively.ConclusionsPP2Ainhibitors play a role in preventing pancreatic cancer through PP2Acα/IKKα/IκBα/p65 pathway.
2.Expression and biological function of miR-622 in gastric cancer cells
Ruizheng MIAO ; Leping LI ; Jianfeng LI ; Meng WANG ; Hongguang SHENG ; Tao XU
Chinese Journal of General Surgery 2011;26(11):932-935
Objective To investigate the biological function of miR-622 in human gastric cancer cell lines of SGC-7901 and NCI-N87 cells and its role in gastric carcinogenesis.Methods We analyzed the expression of miR-622 in those human gastric cancer cell lines by quantitative real-time polymerase chain reaction.Tumorigenesis,migration and invasion ability of miR-622 overexpression was assessed in vitro with miR-622 precursor and inhibitor in in SGC-7901 and NCI-N87 cells.Results The expression level of miR-622 in SGC 7901 was 1.29 ± 0.57,and it was 10.96 ± 1.02 in NCI-N87 cells.The soft agar colony formation rate was 76% in SGC-7901 after transfecting miR-331-3p precursor,the ability of scratch healing was ( 11 ±7) μm,and the ability of transwell invasion was (731 ±3),compared with that in control group,the differences were statistically significant ( P < 0.05 ).Conclusions Over-expression of miR-622 promotes tumorigenesis,migration,and invasion in gastric cancer cells in vitro.
3.Low-temperature deposition manufacturing and property of polylactic-co-glycolic acid composite scaffolds at different ratios
Tao HU ; Mingen XU ; Ming YAN ; Miao GUO ; Ran SHI ; Qingqing ZHOU
Chinese Journal of Tissue Engineering Research 2015;(12):1805-1811
BACKGROUND:The study confirmed that adding tricalcium phosphate or pearl powder in polylactic-co-glycolic acid can complement the performance of both, which provides a good environment for cels and makes a faster and better growth of cels. OBJECTIVE:We used polylactic-co-glycolic acid as matrix, composited with pearl powder or tricalcium phosphate to prepare scaffolds by low-temperature deposition manufacturing. METHODS:Low-temperature deposition manufacturing was utilized to prepare composite scaffold of polylactic-co-glycolic acid/pearl powder or polylactic-co-glycolic acid/tricalcium phosphate at the ratio of 10:0, 5:2, 7:3 and 6:4. Microstructure, contact angle and compression modulus of elasticity of scaffolds were detected. MC3T3-E1 cels basicaly fused at 1×104/cm3 were seeded in the pure nonporous polylactic-co-glycolic acid scaffold, pure polylactic-co-glycolic acid scaffold with holes, polylactic-co-glycolic acid/pearl powder at 5:2 and polylactic-co-glycolic acid/tricalcium phosphate at 5:2 separately for 1 and 3 hours. Cel adhesion rate was detected using flow cytometry. After incubation for 1, 4 and 7 days, cel proliferation was measured using Alamar Blue method. RESULTS AND CONCLUSION:Pure polylactic-co-glycolic acid scaffold had cross-linked microporous structure, with pore size of 3-15 μm. Scaffolds ofpolylactic-co-glycolic acid/pearl powder at 5:2 or polylactic-co-glycolic acid/tricalcium phosphate at 5:2 had good continuous porous structure, with pore size of 10-25 μm. With increased content of pearl powder or tricalcium phosphate, the hydrophilicity of the composite scaffold increased. The addition of pearl powder or tricalcium phosphate could elevate compressive mechanical properties of the composite scaffold. With increased content, the mechanical property of the scaffold enhanced and then reduced. The addition of pearl powder or tricalcium phosphate improved the celular affinity of polylactic-co-glycolic acid and the biocompatibility of the scaffold. The biocompatibility of polylactic-co-glycolic acid/pearl powder scaffold at 5:2 was the best.
4.Identification and screening the mimic epitopes of human Rh(D)blood type antigens
Maohong BIAN ; Jijia SHEN ; Miao LIU ; Wei XU ; Peng YANG ; Shujun LIU ; Tao ZHONG
Chinese Journal of Laboratory Medicine 2008;31(3):305-308
Objective To screen the mimic epitopes of Rh(D)blood group antigens and identify their immunity from phage display peptide library.Methods A twelve mer phage peptide library was biopanned with anti-Rh(D)monoclonal antibody immobilized on plastic surface.After three round panning,thirty-five clones were randomly selected and positive clones were identified by ELISA and cross-reaction,followed by antibody competition inhibition assay and DNA sequencing to obtain the mimic epitopes of Rh (D)blood type antigens.The target phage clones were characterized and the antigenicity was analyzed by Western blot.Results After the third round screening,phages were enriched,and eleven positive clones were obtained.According to sequencing and competition inhibition analysis,the same"-WP-Q-"structure existed in seven of the eleven clones,and they had more than 40%inhibition ratio.The other clones had no same characteristics with low inhibition ratio possibly due to non-specific binding.Western blot analysis indicated that these phage clones could be specifically recognized by the anti-Rh(D)serum and they shared the same antigenicity of Rh(D)protein.Conclusions Rh(D)mimotope of"-WP-Q-"structure is successfully obtained by phage peptide library screening with anti-Rh(D)monoelonal antibody.The results lay the foundation for further exploration of pathogenesis and vaccine development of Rh(D)hemolytic diseases of newborn.
5.Application of hyperinsulinemic euglycemic clamp in the diagnosis and treatment of newly-onset diabetic patients
Tao YUAN ; Weigang ZHAO ; Yong FU ; Yingyue DONG ; Lian DUAN ; Yan JIANG ; Ou WANG ; Miao YU ; Jianping XU
Chinese Journal of Clinical Nutrition 2010;18(2):77-79
Objective To evaluate the application value of hyperinsulinemic euglycemic clamp in the diagnosis and treatment of newly-onset diabetic patients. Methods Totally 11 newly-onset diabetic inpatients (10 patients with type 2 diabetes mellitus and 1 patient with latent autoimmune diabetes of adulthood) who were diagnosed in the Department of Endocrinology of Peking Union Medical College Hospital from January 2009 to August 2009 were included in this study. Hyperinsulinemic euglycemic clamp was applied to measure the glucose disposal rate (M value). Afterwards insulin pump therapy was applied and the total insulin dosage per day to get to the target of the fasting and postprandial blood glucose was calculated. Final]y the relationships between insulin dosage per day and the M value, body mass index (BMI) , fasting blood glucose, fasting insulin level were separately analyzed. Results The insulin dosage was only negatively correlated with M value (r = - 0. 83, P = 0. 003), and was not significantly correlated with BMI (r = 0.54, P = 0.106), fasting blood glucose (r = - 0. 16, P =0. 657) , and fasting insulin (r = 0. 16, P = 0. 659). The formula of insulin dosage and M value according to the mathematic model as follows: insulin dosage per day = - 3. 327 M + 49. 849. Conclusion Hyperinsulinemic euglycemic clamp can effectively evaluate the insulin sensitivity in the newly-onset type 2 diabetic patients, and thus can be a useful tool in deciding the clinical insulin dosage.
6.Cantharidin induces apoptosis in pancreatic cancer cell lines PANC1 and CFPAC-1
Wei LI ; Zheng CHEN ; Yang ZONG ; Feiran GONG ; Yi ZHU ; Hong YIN ; Zekuan XU ; Min TAO ; Yi MIAO
Chinese Journal of Pancreatology 2011;11(4):255-258
Objective To investigate the apoptosis induction effect of Cantharidin on pancreatic cancer cell line PANC1 and CFPAC-1 and possible mechanism. Methods PANC1 and CFPAC-1 was treated with Cantharidin. Cell growth was determined by MTT. Apoptosis was measured by flow cytometry. Caspase activity was measured by using enzyme chemical method. Apoptosis-related gene expressions were determined by using RT-PCR and Western blotting. Results Cantharidin significantly inhibited the growth of pancreatic cancer cells PANC1, CFPAC-1 and induced apoptosis in a dose-dependent manner. Seventy-two hours after 10 μmol/L Cantharidin treatment, the inhibitory rates of PANC1, CFPAC-1 were (52.95 ± 6.34)% and (71.21 ±6.30)%. Twenty-four hours after treatment, the early and later period apoptotic cell of PANC1 was increased from 7.35% to 24.89%, from 6.36% to 17.73%. The early and later period apoptotic cell of CFPAC was increased from 6.39% to 24.70%, from 9.21% to 12.58% (P<0.01). Activity of caspase 8 and caspase 9 in PANC1 cells was (155.8 + 11.5)% and (194.6 ± 14.7)% when compared with that of control group. Activity of caspase 8 and caspase 9 in CFPAC- 1 was ( 182.5 ± 24.3 ) % and ( 215.8 ± 12.2) %when compared with that of control group ( P < 0. 01 ). The expression of pro-apoptotic genes, TNF-α,TRAILR1, TRAILR2, Bad, Bak and Bid was elevated, the expression of anti-apoptotic Bcl-2 gene was decreased. Conclusions Cantharidin can induce apoptosis in pancreatic cancer cell lines by activating caspase,up-regulating the expression of pro-apoptotic genes and down-regulating the expression of anti-apoptotic genes.
7.Anti-CD25 monoclonal antibody as a salvage therapy for steroid-refractory acute graft-versus-host disease in 80 patients receiving allogeneic hematopoietic stem cell transplantation
Tao TAO ; Shengli XUE ; Feng CHEN ; Yang XU ; Xiao MA ; Miao MIAO ; Xiaowen TANG ; Depei WU
Chinese Journal of Internal Medicine 2018;57(5):324-329
Objective To investigate the efficacy of anti-CD25 monoclonal antibody for steroid-refractory acute graft-versus-host disease (SR-aGVHD) in allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients.Methods A total of 80 patients with SR-aGVHD from January 1st 2012 to December 31st 2016 were enrolled in this study.Acute GVHD were classified as classic aGVHD (n=72) and late-onset aGVHD (n=8).Anti-CD25 monoclonal antibodys (mAb) were administrated on days 1,4,8,15,and 22.The efficacy of anti-CD25 mAb was evaluated at day 28 after the initial treatment.The associated factors of clinical outcome were analyzed.Results The overall response (OR) rate of anti-CD25 mAb was 75% (60/80),with complete response (CR) rate,partial response (PR) rate and no response(NR) rate 52.5% (42/80),22.5% (18/80),and 25% (20/80),respectively.GVHD-relapse was not observed with a median follow-up time of 394.5 days (range,12-1 761 days).The 6-month overall survival (OS) rate was 68.4% (95%CI 63.2%-73.6%).The 1-year OS rate was 63.1% (95%CI 57.6%-68.6%),and 2-years OS rate was 50.7% (95%CI 44.3%-57.1%).Non-relapse mortality (NRM) rate of 1 and 3 years was 32.6% (95%CI 27.2%-38%) and 41.7% (95%CI 35.3%-48.1%),respectively.The 1 and 2 years cumulative incidence of chronic graft versus host disease (cGVHD) was 32.9% (95%CI 26.4%-39.4%) and 38.9% (95%CI 31.8%-46.0%).By univariate and multivariate analysis,liver involvcment was an independent poor risk factor of SR-aGVHD (OR=4.66,95% CI 1.145-18.962,P=0.032).Conclusion Anti-CD25 mAb serves as an alternative and effective salvage therapy for SR-aGVHD at present.Liver involvement is a predictive factor of poor response in patients with SR-aGVHD.
8.Microsurgical anatomy of white matter fiber tracts and important structures of the temporal lobe
Jin XU ; Feng WANG ; Fa-Zheng SHEN ; Lin MIAO ; Yong-Long XU ; Tao SUN
Chinese Journal of Neuromedicine 2012;11(12):1233-1237
Objective To investigate the microsurgical anatomy of white matter fiber tracts and the important structures of the temporal lobe,and analyze its functional and clinical implications.Methods Ten formalin (100 g/L)-fixed human brain hemispheres were dissected using the Klingler fiber dissection technique,with the aid of an operating microscope at 4-25 magnification; the microsurgical anatomy of white matter fiber tracts and the important structures of the temporal lobe was observed.Results In the temporal lobe,a large number of complex white matter fiber tracts were noted locating lateral to the lateral wall of the temporal horn of lateral ventricle and superior to the roof wall.The vertical segment of the superior longitudinal fasciculus,occipitotemporopontine tract,inferior occipitofrontal fascicle,anterior and middle tracts of the optic radiation were located from lateral to the lateral wall of the temporal hom in turn; claustro-opercular and zinsulo-opercular fibers of the external and extreme capsules,auditory radiations,uncinate fasciculus,part of occipitotemporopontine tract,part of inferior occipitofrontal fascicle,anterior commissure,anterior and middle tracts of the optic radiation(including Meyer's loop),the ansa peduncularis and the stria terminalis were located from superior to inferior to the lateral wall of the temporal horn in turn.The lateral wall of the temporal horn of the lateral ventricle was composed of corpus callosum radiation,whose roof wall composed of the tail of caudate nucleus.Furthermore,amygdala composed of the anterior,tip and medial wall of the temporal horn,and the hippocampus constituted the medial wall of the temporal horn.The cerebral foot loop was an important medial structure of the temporal horn.Conclusion It's important in the clinical diagnosis and treatment to improve the knowledge and understanding of white matter fiber tracts and important structures of the temporal lobe.
9.Depyrogenation in key manufacturing processes of Reduning injection.
Miao LI ; Yuling XU ; Juan SONG ; Yongxiang WANG ; Yingzhi PAN ; Zhengzhong WANG ; Wei XIAO ; Tao LIU
China Journal of Chinese Materia Medica 2011;36(6):663-665
OBJECTIVETo investigate the effect of removing bacterial endotoxin in the key processes of Reduning injection.
METHODThe content of bacterial endotoxins was detected by kenitic-turbidimetry and the removal efficacy was studied before and after using 0.8% of activated carbon and ultrafiltration with molecular weight cut-off of 10 x 10(3).
RESULTThe adsorption rate of bacterial endotoxins was 78.7% by using activated carbon, while the removal efficacy of bacterial endotoxins was 99.6% with ultrafiltration membrane at cut-off molecular weight 10 x 10(3).
CONCLUSIONThe key technology can effectively guarantee the safety of Reduning injection.
Adsorption ; Endotoxins ; isolation & purification ; Injections ; Pyrogens ; isolation & purification ; Ultrafiltration
10.Bootstrapping Data Envelopment Analysis of Efficiency and Productivity of County Public Hospitals in Eastern, Central, and Western China after the Public Hospital Reform
WANG MAN-LI ; FANG HAI-QING ; TAO HONG-BING ; CHENG ZHAO-HUI ; LIN XIAO-JUN ; CAI MIAO ; XU CHANG ; JIANG SHUAI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(5):681-692
China implemented the public hospital reform in 2012.This study utilized bootstrapping data envelopment analysis (DEA) to evaluate the technical efficiency (TE) and productivity of county public hospitals in Eastern,Central,and Western China after the 2012 public hospital reform.Data from 127 county public hospitals (39,45,and 43 in Eastern,Central,and Western China,respectively) were collected during 2012-2015.Changes of TE and productivity over time were estimated by bootstrapping DEA and bootstrapping Malmquist.The disparities in TE and productivity among public hospitals in the three regions of China were compared by Kruskal-Wallis H test and Mann-Whitney U test.The average bias-corrected TE values for the four-year period were 0.6442,0.5785,0.6099,and 0.6094 in Eastern,Central,and Western China,and the entire country respectively,with average non-technical efficiency,low pure technical efficiency (PTE),and high scale efficiency found.Productivity increased by 8.12%,0.25%,12.11%,and 11.58% in China and its three regions during 2012-2015,and such increase in productivity resulted from progressive technological changes by 16.42%,6.32%,21.08%,and 21.42%,respectively.The TE and PTE of the county hospitals significantly differed among the three regions of China.Eastern and Western China showed significantly higher TE and PTE than Central China.More than 60% of county public hospitals in China and its three areas operated at decreasing return scales.There was a considerable space for TE improvement in county hospitals in China and its three regions.During 2012-2015,the hospitals experienced progressive productivity;however,the PTE changed adversely.Moreover,Central China continuously achieved a significantly lower efficiency score than Eastern and Westem China.Decision makers and administrators in China should identify the causes of the observed inefficiencies and take appropriate measures to increase the efficiency of county public hospitals in the three areas of China,especially in Central China.