Objective To investigate whether EZH2 participates in the process of authphagy and its regulatory mechanism in CRC (colorectal cancer) .Methods ZEB1 ,EZH2 and PTEN expression were measured by Western blot and immunohistochemistry respectively .ZEB1 ,EZH2 and PTEN mRNA level were measured by real-time PCR .Electron microscopy was introduced to validate the existence of autophagy .Results Knockdown of EZH2 induced the formation of autophagosome in colorectal cancer cell lines , which was evident on electron microscopy .Furthermore ,Western Blot and real-time PCR data showed that ZEB1 and EZH2 may regulate the expression of PTEN ,which played a vital role in autophagy .Moreover ,downregulation of ZEB1 significantly reduced the expression of EZH2 .An inverse correlation between the expression of EZH2 and ZEB1 ,and the expression of PTEN was also revealed in CRC tissues ,when compared with normal tissue in patients .Conclusion The impact of EZH2 on autophagy via PTEN during CRC carcinogenesis is revealed .At the same time ,EZH2 expression may be regulated by ZEB1 in colorectal cancer .

Background:Vitamin D receptor( VDR)is a member of the steroid hormone receptor superfamily,which plays roles in various biological processes including cell proliferation,differentiation,apoptosis and immune responses,and is low expressed in many malignant tumors. Aims:To evaluate the expression and regulation mechanism of VDR in colorectal cancer. Methods:A total of 30 patients with colorectal cancer admitted from February 2010 to December 2012 at Ren Ji Hospital,School of Medicine,Shanghai Jiao Tong University were enrolled. Expression of VDR in cancerous tissue and para-cancer noncancerous tissue was determined by immunohistochemistry. Clinical data of 224 patients with colorectal cancer were collected from Gene Expression Omnibus( GEO)for analyzing the correlation between VDR expression in cancerous tissue and clinicopathological characteristics as well as survival time. Expression of VDR in human colon cancer cell lines HCT116 and SW620 transfected with enhancer of zeste homolog 2( EZH2 )-siRNA or treated with 5-azacytidine (5-AZA)was determined by real-time PCR,and methylation level of VDR promoter was determined by methylation-specific PCR( MSP). Results:Positivity rate of VDR was significantly lower in colorectal cancer tissue than that in para-cancer noncancerous tissue( 26. 7% vs. 70. 0%,P < 0. 001 ). Expression of VDR was negatively correlated with histological staging,distant metastasis,vascular metastasis and lymph node metastasis of colorectal cancer(P<0. 05). Survival time was significantly longer in patients with high expression of VDR than patients with low expression of VDR (P=0. 032). Compared with cells transfected with control-siRNA,expression of EZH2 mRNA and methylation level of VDR promoter in HCT116 and SW620 cells transfected with EZH2-siRNA were significantly decreased(P<0. 05),and expression of VDR mRNA was significantly increased(P<0. 05). Compared with negative control group,methylation level of VDR promoter in HCT116 and SW620 cells treated with 5-AZA was significantly decreased(P<0. 05),and expression of VDR mRNA was significantly increased(P<0. 05). Conclusions:Expression of VDR in colorectal cancer is down-regulated and positively correlated with a favourable prognosis. Transcriptional repression of VDR in colorectal cancer is co-regulated by histone methylation and DNA methylation.

Growth hormone (GHRH) and pitutary adenylate cyclase activating peptide (PACAP) are the members of the PACAP/Glucagon superfamily,who are related in both sequence and function.Their stimulation of GH secretion and animal growth is concerned.A series of expression plasmid,pIRES1-GHRH-PACAP (P-G-P),plRESI-GHRH (P-G) and plRESI-PACAP(P-P),were constructed,extracted and purified,then transfected into CHO cell line with Lipofectamine.The expression was examined by RT-PCR,dot-ELISA and Western blotting.The biological activity of expression products was detected in rats.At 8 h after injection of transfection supematant,serum IGF-I concentrations in P-G-P group were significantly higher than that in other groups(P ＜ 0.05).PLGA encapsulating plasmid microspheres were prepared and injected intramuscularly into rabbit legs.Growth behavior and IGF-1 level were measured at day 0,15,30 and 45 after injection.Greater body weights gain and higher serum 1GF- [ levels were observed in three plasmid microsphere injection groups,compared with control group.At day 30,the body weight gain in P-G-P group was greater than saline group (81%,P＜ 0.01),P-G mierosphere group (15%,P＜ 0.05) and P-P microsphere group (7%,P＞ 0.05),serum IGF-I concentration in P-G-P microsphere group showed a 16.68% increase to P-G microsphere (P ＞ 0.05),a 17.14% increase to P-P microsphere(P ＞ 0.05) and a 50.46% increase to control (P ＜ 0.05).These results suggest that co-expression of GHRH and PACAP in one expression plasmid might exert an additive stimulation of GH secretion and growth when delivered into rabbit skeletal muscle with PLGA mierosphere.The results may provide a new approach to regulate animal growth.

Objective To investigate effects of capsaicin on proliferation,migration and invasion of human large cell carcinoma NCI-H460 and discuss the possible mechanisms by which capsaicin inhibits non-small cell lung cancer. Methods NCI-H460 cells were cultured in vitro and treated with capsaicin at various concentrations. Colony formation assay,wound healing assay,and transwell chamber invasion assay were used to detect the effects of capsaicin on proliferation,migration and invasion of NCI-H460 cells. Western blotting was used to detect the protein expression of E-cadherin,N-cadherin,Vimentin and Snail. Results The rate of colony formation of 100,200,300 μmol.L-1 of capsaicin were (91.17±24.38)%,(43.22±12.91)% and (15.71±4.26)%,respectively,the control group was (99.53±21.25)%.the rate of migration of NCI-H460 cells of 25,50, 100 μmol.L-1 of capsaicin were (85.83±17.43)%,(37.92±10.16)% and(21.08±6.39)%,respectively,the control group was (93.04±20.23)%.The number of invasion of NCI-H460 cells of 25,50,100 μmol.L-1 of capsaicin were (99±18.2),(75± 17.9) and( 52 ± 14. 1 ) , respectively, the control group was ( 107 ± 20. 4 ) . The middle and high-dose capsaicincould inhibit proliferation,migration and invasion of NCI-H460 cells ( P<0.05) ,and E-cadherin expression level was significantly up-regulated and N-cadherin, Vimentin, Snail expression level was significantly down-regulated ( P<0. 05 ) . Conclusion Capsaicin can inhibit proliferation,migration and invasion of NCI-H460 cells,the mechanism may be related to up-regulation of E-cadherin and down-regulation of N-cadherin,Vimentin and Snail expression levels.

Objective:To establish and evaluate a newly established method of enzyme-linked immu-nosorbent assay (ELISA) for measuring human autoantibody to folate receptor (FR).Methods: Folate receptor was extracted and purified from healthy woman placenta tissues .The protein was coated on 96-well plates.Goat monoclonal antibody was used as detecting antibody to set up the indirect ELISA proce -dure.The sensitivity, precision and linearity of the method were evaluated .Further, the method was compared with the ELISA method with commercialized bovine folate binding protein ( FBP) by determi-ning autoantibody levels in 24 individuals .Results:The measuring range of the standard curve was from 6 .25 ×10 -4 to 8 ×10 -2 ( the IgG concentration of pooled plasma from healthy donors was defined as 1 ) . The lowest detectable level was 3.13 ×10 -4 .The intra-and inter-assay coefficients of variations were 2.74%-8.07% and 4.16% -8.23%, respectively.Linearity test results were considered within acceptable limits.The data from FBP-ELISA and FR-ELISA were highly correlated ( r=0.954, P <0.001);The value from FR-ELISA was higher by 14% than that from FBP-ELISA.Conclusion: The ELISA method for measuring human autoantibody IgG to folate receptor was successfully established using human FR as coating protein .The method is sensitive and repeatable and can be used in large-scale population study .

Objective To investigate the efficacy and safety of Lipo-prostaglandin E1 (LipoPGE1) in the treatment of non-ST segment elevation acute myocardial infarction (NSTEAMI).Methods A total of 86 patients with NSTEAMI were randomly and equally divided into LipoPGE1 group (n= 43) which received intravenous LipoPGE1 combined with low-molecular-weight heparin,aspirin, clopidogrel and other basic therapy, and the control group (n=43) which received placebo combined with the same therapy. The basic clinical settings, curative effect, main adverse cardiovascular events (MACEs) within 30 days including sudden death, new-onset myocardial infarction and target vessel revascularization, bleeding complications and drug adverse effects were observed. Results There were no significant differences in basic clinical characteristics between the two groups. Compared with control group, the patients in LipoPGE1 group showed. significant improvements of ECG (93.0% vs. 74.4%), angina (95.3% vs. 81.4%, both P＜0. 05), the incidences of left heart failure (2.3% vs. 14.0%) and MACEs within 30 days (4.7% vs.18.6%)(both P＜0.05). There were no serious drug adverse effects. Conclusions The LipoPGE1 combined with heparin, aspirin and clopidogrel is effective and safe in the treatment of NSTEAMI,which could improve the clinical symptoms, distal myocardium perfusion and cardiac function,decrease the incidence of MACEs.

The data of 10 cases of traumatic extra-articular ankylosis of temporomandibular joint(TMJ),including the type of trauma and the type of ankylosis,pathology,treatment method,prognosis,and so on were collected and analyzed.A reference of diagnosis and treatment is provided.

Objective To analyze and discuss four methods of calculating likelihood ratio of DNA mixture. Methods In the case with CNAS-T0757 proficiency testing in 2013, the likelihood ratios were calculated and compared among four methods, including unrestricted combinatorial method, Clayton’s method, p2 principle method, and recommendations fromISFG . Results The likelihood ratios were maximumby Clayton’s method and recommendations fromISFG , followed by result of the unrestricted combinational method. The minimu mlikelihood ratio was obtained by p2 principle. Conclusion The unrestricted combi-national method could give furthest consideration to both information preservation and appraiser protection.

Objective To explore the variation trends of interleukin-1β( IL-1β) and intercellular adhesion molecule-1 (ICAM-1) in both normal and affected sides of brain tissues in rats with ischemia-reperfusion injury and the therapeutic action of electroacupuncture.Methods The cerebral ischemia-reperfusion model was established with suture embolization in the right middle cerebral artery.The rats were randomly divided into control group, model group and electroacupunture group.Each group was then divided into six subgroups by the time after operation (12 h,24 h,48 h,72 h,96 h,144 h), ten rats in each subgroup. Frozen sections of brain tissues were prepared and the expression of IL-1βand ICAM-1 in brain tissues of both sides were detec-ted by immunohistochemistry.Results The expressions of IL-1βand ICAM-1 showed typical bimodal pattern in both affected is-chemic region and contralateral normal region.In the model group, the peaks of IL-1βin the cerebral ischemic region were at 12 h and 48 h, while in the contralateral normal region the peaks were at 12 h and 144 h, the expression of IL-1βin the ischemic region was significantly higher than that in the contralateral normal region at 48 h (P<0.05), and lower at 96 h and 144 h (P <0.05).In the electroacupuncture group, the expressions of IL-1βin the ipsilateral region were significantly lower than that in the contralateral region at 24 h, 48 h and 144 h (P<0.05).In the model group, the peaks of ICAM-1 in the cerebral ischemic regions were at 24 h and 72 h, while in the contralateral normal regions the peaks were at 24 h and 144 h.In the electroacupunc-ture group, the expressions of ICAM-1 in the ischemic regions were significantly lower than that in the contralateral normal re-gions at all the 12 h, 24 h, 48 h, 72 h and 144 h (P<0.05).Conclusions Our findings suggest that electroacupuncture may inhibit the inflammation of ischemia/reperfusion brain tissue through reducing the expression of IL-1βand ICAM-1 to relieve the cerebral ischemia-reperfusion injury.

Objective To investigate the effect of JUC on the incisions for episiotomy.Methods Three hundred primiparas to undergo episiotomy in our hospital were divided into two groups in equal number.The experiment group was given JUC Spray before suturing and the control group did not use any solution.In the two groups,antibiotics were not used after the operation,and the incisions were only cleaned with 0.5%povidone-iodine 2 times a day.Result There were significant differences between the two groups in terms of postoperative pains,inflammation and healing in the wounds,and hospital stay(P<0.05).Conclusions Application of JUC after episiotomy could be long-acting in antibacteria.It can reduce wound pain,improve wound healing rate, decrease the medical cost and shorten the hospital stay.