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.

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.

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.

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.

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.

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.

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.

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.

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.

AIM:To investigate the effect of ischemic postconditioning ( IPC) on autophagy induced by focal cerebral ischemia reperfusion ( I/R) in rats.METHODS:Healthy male SD rats were assigned randomly into sham-opera-tion (sham) group, I/R group and IPC group with 10 rats in each group.The rats in sham group were only exposed the right common , internal and external carotid artery surgically .The rats in I/R group were subjected to right middle cerebral artery occlusion (MCAO) by the modified Longa suture method for 2 h followed by 24 h of reperfusion.The rats in IPC group were subjected to MCAO for 2 h followed by reperfusion of the ipsilateral common carotid artery occlusion for 10 s for 5 episodes, and then reperfusion for 24 h.Autophagy was obeserved by transmission electron microscopy (TEM).The pro-tein levels of mammalian target of rapamycin (mTOR), p-mTOR and microtubule associated protein light chain 3 (LC3)-II in brain tissue of the rats were determined by Western blot .Pathological changes of brain tissue were observed by HE staining.RESULTS:The protein levels of mTOR and p-mTOR in IPC group were significantly higher than those in I/R group (P<0.05).The expression of LC3-II in IPC group was significantly lower than that in I/R group (P<0.01).The cerebral infarction area and brain water content in IPC group were significantly lower than those in I /R group (P<0.01). HE staining showed that neurons degeneration and necrosis in IPC group were significantly alleviated compared with I /R group.TEM observation showed that IPC revealed fewer autophagosomes , with much less severe cell damage than that in I/R group.CONCLUSION:IPC reduces brain ischemia reperfusion damage by decreasing autophagy of brain cells , which might be related to the activation of mTOR .