Objective To investigate the effects of transfection of chemokine receptor CXCR4 antisense-RNA on the functional expression of VEGF-C mRNA and the invasive ability of colon cell line HT-29 in vitro.Methods PCR primers were designed according to the coding sequence of CXCR4 gene.EcoR Ⅰ and BamH Ⅰ recognition sequences and cutting sites were added to the 5' end of the antisense primer.The purified PCR fragment was retro-inserted into the cloning site of eukaryotic expression vector pcDNA3.1(+).The pcDNA3.1(+)for CXCR4 antisense-RNA was transfected into colon cancer HT-29 cells by liposome transfection.The expression of VEGF-C mRNA was detected by RT-PCR.The expression of CXCR4 protein was determined by Western blot.At the same time,cell growth kinetics was assessed by MTT assay,and in vitro invasive ability was assessed with Boyden chamber.Results CXCR4 antisense-RNA recombinant was successfully constructed.The expression of VEGF-C mRNA in antisense-CXCR4 transfected(HT-29tran)group decreased by 54.2%,while the expression of VEGF-C mRNA in vacant-transfected(HT-29KZ)group only decreased by 9.4% campared with non-transfected(HT-29)group.The difference between the two groups mentioned above was remarkable(P

Objective To investigate the molecular mechanism of metformin. Methods The changes of protein expression of IRS-1 in liver, skeletal muscle and adipose tissues after treatment with metformin for OLETF rats, a model of spontaneous type 2 diabetes mellitus, were measured by Western blot analysis, and compared with those before treatment. Results 22 weeks after the treatment with metformin, the protein expression of IRS-1 was significantly increased in liver (P0. 05) ;and the protein expression of IRS-1 in adipose tissue was significantly decreased (P

Objective To study the influence of unilateral and bilateral amplification on the effect of hearing aid evaluation .Methods Using the subjective method that International Outcome Inventory for Hearing Aids (IOI-HA) and objective method that medium acoustic intensity (65 dB SPL) word recognition score(WRS) to evaluate the effect of unilateral and bilateral hearing aid fitting of middle -aged severe sensorneural hearing loss .Results Hearing aid were used for severe sensorneural hearing loss and the improvement of monosyllables and sentences in quiet and noise test of unilateral were 35 .73% ,43 .15% ,43 .23% ;the improvement of monosyllables and sentences in quiet and noise test of bilateral were 37 .90% ,51 .33% ,54 .86% .The WRS of bilateral was higher than unilater-al .The score of IOI-HA was 15~37 ,meaning patients with severe sensorneural hearing loss were satisfied with hearing aid ,and there was no statistical significance between unilateral and bilateral fitting .Conclusion The bilat-eral hearing aid fitting was better than unilateral .Binaural hearing loss are recommended to fit bilaleral hearing aids .

Objective To analyze the metabolic profile of children with dyskinetic cerebral palsy by metabolomics, and its abnormal metabolic pathway. Methods The serum of 10 children with dyskinetic cerebral palsy (patient group) and 7 healthy children (control group) aged 6 to 12 years were collected at clinic from May to August, 2014. The serum samples were tested by the nuclear magnetic resonance spectrometer and the spectroscopies were discriminated by partial least squares-discriminant analysis. According to the human metabolome database, the final metabolites disturbed would be figured out. Results 15 chemical shifts were defined, and 6 of them, including 2.04 ppm, 2.12 ppm, 3.00 ppm, 3.24 ppm, 3.76 ppm, 6.50 ppm, were significantly different between 2 groups (P<0.05). The KEGG Pathway Database showed that the levels of taurine, fumarate, oxaloacete, pyruvate, citrate, aspartate, succinate, malate, cysteine decreased, and the levels of glutamate, 2-oxoglutarate, glutamine, leucine, alanine increased. The abnormal metabolism was found in taurine metabolism, glutamine me-tabolism and energy metabolism pathways. Conclusion Based on metabolomics, the metabolic profile of children with dyskinetic cerebral palsy was discriminated out successfully. The further research can focus on the small molecules found out.

Objective To investigate the risk factors for early postoperative cognitive function in the patients with peritoneal surface malignancies after cytoreductive surgery and hyperthermic intra-peritoneal chemotherapy(CRS-HIPEC).Methods Fifty-one patients(21 men and 30 women), ranged from 25 to 65 years,42-80 kg,ASA Ⅰ-Ⅲ,undergoing CRS-HIPEC under combined intrave-nous-inhalational anesthesia,were studied.Patients were assigned into postoperative cognitive dys-function (POCD)group or non-POCD group according to their performances of visual verbal learning test,concept shifting task,letter-digit coding test and stroop color-word test 1 day before operation and 7 days after operation.Years of education,medical history,duration of operation,intraoperative blood loss,frequency of cardiovascular events,amount of fluid infused per hour and VAS scores were recorded.Venous blood samples were taken at five time points:before surgery(T0 ),30 minutes after the beginning of the procedure(T1 ),30 minutes after the beginning of HIPEC(T2 ),at the end of the surgery(T3 )and 24 hours after the surgery(T4 ),to determine the concentration of serum amyloid A (SAA).pH,PaCO 2, Hb,blood glucose were recorded at T0-T2.Then the data was statistically analyzed.Results According to the diagnostic criteria,twenty patients developed POCD 20 (39.2%). There were significant differences between POCD and non-POCD groups on age,gender, pre-operative complications and the origin of tumor(P <0.05).The concentration of SAA increased from T2 and reached the peak at T4 ,and SAA concentration for patients in POCD group was higher than that for patients in non-POCD group(P <0.05).Compared with non-POCD group,the levels of blood glucose were significantly increased in POCD group at T2 (P <0.05).Conclusion CRS-HIPEC resul-ted in exaggerated and prolonged inflammatory response.Advanced age,female,diabetes,hyperten-sion,peritoneal carcinomatosis from ovarian cancer and endometrial cancer are associated with early POCD in the patients undergoing CRS-HIPEC.

Objective To investigate the relationship between the mechanism of ischemic postconditioning-induced activation of nuclear factor-E2 related factor 2 (Nrf2)-antioxidant response element (ARE) signaling pathway during myocardial ischemia-reperfusion (I/R) and reactive oxygen species (ROS).Methods Healthy male Sprague-Dawley rats, aged 16-20 weeks, weighing 250-300 g, were heparinized and anesthetized with intraperitoneal 1％ pentobarbital sodium 40 mg/kg.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution.Thirty-two isolated rat hearts were randomly divided into 4 groups (n=8 each) using a random number table: control group (group C) , group I/R,ischemic postconditioning group (group IPO) , and N-(2-mercaptopropionyl)-glycine (a ROS scavenger) + IPO group (group M + IPO).After 20 min of equilibration, group C was continuously perfused with K-H solution for 100 min, and the isolated hearts received the drugs via the perfusion system in the other groups.Group I/R was perfused with cardioplegic solution 4 ℃ St.Thomas, and then was subjected to 40 min of ischemia at 32 ℃ followed by 60 min of reperfusion.In group IPO, ischemic postconditioning was induced by 6 cycles of 10 s reperfusion followed by 10 s limb ischemia starting from the onset of reperfusion, and the hearts were then perfused for 58 min.In group M + IPO, the hearts were perfused with K-H solution containing N-(2-mercaptopropionyl)-glycine 2 m mol/L for 3 min starting from the onset of reperfusion,underwent 2 min of ischemic postconditioning, and then was perfused for 55 min.Heart rate (HR), left ventricular developed pressure (LVDP), left ventricular end-diastolic pressure (LVEDP),and positive maximal pressure of left ventricular increase (+dp/dtmax) were recorded at the end of equilibration and of reperfusion.At 5 min of reperfusion and the end of reperfusion, myocardial specimens were obtained from the left ventricle for determination of ROS content by enzyme-linked immunosorbent assay.At the end of reperfusion, myocardial specimens were obtained from the left ventricle for examination of the ultrastructure of myocardial cells and for determination of Nrf2, heme oxygenase-1 (HO-1) , quinone oxidoreductase 1 (NQO1), and superoxide dismutase 1 (SOD1) mRNA and protein expression (by using Western blot and real-time polymerase chain reaction).The damage to myocardial mitochondria was assessed using Flameng scoring.Results Compared with group C, HR, +dp/dtmax and LVDP were significantly decreased, and LVEDP was increased at the end of reperfusion in I/R and M+IPO groups, HR and LVDP were decreased, LVEDP was increased, and no significant changes were found in +dp/dtmax at the end of reperfusion in IPO group, Flameng score was increased in I/R, IPO and M+IPO groups , the ROS content was increased at the end of reperfusion in I/R, IPO and M+IPO groups, and Nrf2, HO-1,NQO1 and SOD1 mRNA and protein expression was down-regulated at the end of reperfusion in I/R, IPO and M+IPO groups.Compared with group I/R, HR, +dp/dtmax and LVDP were significantly increased, and LVEDP and ROS content were decreased at the end of reperfusion, Nrf2, HO-1, NQO1 and SOD1 mRNA and protein expression was up-regulated at the end of reperfusion in IPO and M+IPO groups, Flameng score was decreased in IPO group, there was no significant change in Flameng score in M+IPO group.Compared with group IPO, HR, +dp/dtmax and LVDP were significantly decreased, LVEDP and ROS content were increased at the end of reperfusion, Flameng score was increased, and Nrf2, HO-1, NQO1 and SOD1 mRNA and protein expression was down-regulated in M+IPO group.Conclusion Ischemic postconditioning can regulate ROS level and activate Nrf2-ARE signaling pathway, thus attenuating myocardial I/R injury in rats.

Objective To observe the activation mechanism of Nrf2-ARE pathway in protective effect of ischemia and pinacidil postconditioning on isolated rat hearts.Methods The hearts of adult male Sprague Dawley rats were established ischemia-reperfusion injury model,and devided into six groups(n =8,each group),i.e.Normal group(Group N),ischemiareperfusion group (Group Con,I/R),ischemic postconditioning group (Group IPO),pinacidil postconditioning group (Group P50),N-(2-mercaptopropionyl)-glycine(MPG,2mmol/L) + IPO group(Group M + IPO),MPG + P50 group(Group M + P50).Rat hearts were perfused with Krebs-Henseleit(K-H) buffer for 20 minutes for equilibration.Subsequently,Group N was perfused with K-H buffer for 100 minutes after equilibration,Group Con was perfused with 4℃ ST.Thomas solution to stop the heart beating after equilibration,then the hearts were underwent 40 minutes global ischemia under 32℃,and followed by the K-H solution for 60 minutes.Group IPO after global ischemia period,the hearts were subjected to six 10-seconds cycles of ischemia/reperfusion at the beginning of reperfusion,then were reperfused for 58 minutes.Group P50 after global ischemia,rat hearts were perfused with K-H buffer containing pinacidil(50.μmol/L) for 2 minutes before reperfusion.Group M + IPO after global ischemia,the hearts were subjected to perfuse with K-H buffer containing MPG(2 mmol/L) for 3 minutes,and then underwent six 10-seconds cycles of ischemia/reperfusion before reperfusion.Group M + P50 after global ischemia,the hearts were perfused with K-H buffer containing MPG(2 mmol/L) for 3 minutes,and then subjected to perfuse with K-H buffer containing pinacidil(50 μmol/L) for 2 minutes before reperfusion.Cardiac function indexes(such as HR,LVDP,LVEDP,and the Max dp/dt) at the end point of equilibration and repeffusion were observed and recorded.The ultrastructure of myocardial tissue was observed by electron microscopy and the mitochondrial Flameng score was calculated.RT-PCR and western-blot were applied to detect the gene transcription and protein expression of HO-1,NQO1,SOD1,and Nrf2 in left ventricular myocardial tissue after reperfusion.Results The HR,LVDP and + dp/dtmax at the end of reperfusion:the cardiac function indexes are lower among each group compared with group N,group 1PO and group P50 are better than group Con (P ＜ 0.05).Compared with group IPO,there is no significant difference in group group P50,but group M + IPO is obviously decreased(P ＜ 0.05).Compared with group P50,group M + P50 index is decreased significantly(P ＜ 0.05).The LVEDP at the end of reperfusion is lower than that among each group as compared with group Con,which is significantly increased in group Con (P ＜ 0.05).Compared with group IPO,there is no significant difference in group P50,but group M + IPO is significantly increased(P ＜ 0.05).Compared with group P50,the group M + P50 is obviously decreased(P ＜ 0.05).The ultrastructure of myocardial tissue in group N is mostly normal,group Con presence serious damage.The ultrastructure damage of myocardial tissue is improved in group IPO and group P50 as compared with that in group Con,while group M + IPO is more serious than group IPO,group M + P50 is more serious group P50.The mitochondrial Flameng score is higher among each group as compared with group N (P ＜ 0.05),the score is lower in group IPO and group P50 as compared with group Con and corresponding nonblocking group (M + IPO,M + P50,P ＜0.05).The mRNA and the protein expressions of HO-1,NQO1,SOD1 and Nrf2 among each group are lower as compared with group N(P ＜0.05).Compared with those in group Con,the mRNA and the protein expressions in group IPO and group P50 are obviously increased(P ＜ 0.05),group IPO and group P50 are higher than those in group adding active oxygen scavenger(MPG) (P ＜ 0.05).Conclusion Ischemic postconditioning and pinacidil postconditioning have protective effect of myocardial tissue from ischemia reperfusion injury,while improve the cardiac function index.The cardiac protective effect of Ischemic and Pinacidil postconditioning methods may be involved the ROS in early reperfusion,which activate the Nrf2-ARE pathway,and up-regulate the expression downstream antioxidant protein and phase Ⅱ detoxifying enzyme,ultimately improve the cardiac function index during the reperfusion period.

Objective To evaluate the value of retrospective ECG-gated 256 slices spiral CT technique in the scanning of inferior phrenic artery (IPA).Methods 80 patients with underdone abdominal CTA were divided randomly into two groups (40 patients each)as regular abdominal CTA scanning group and retrospective ECG-gated technique group.And further analysis was done to evaluate the display of IPA branches and the image quality.Results 80 patients were all scanned successfully.40 cases LIPA and 40 cases RIPA were showed in the regular group.80 cases showed 1st level branches of IPA,73 cases with 2nd level,59 cases with 3rd and 43 ca-ses showing 4th level.40 cases LIPA and 40 cases RIPA were showed in ECG-gated group.And the numbers of IPA branches levels were 80,79,71,65,respectively.There is no significant difference in the ability of showing the1st level IPA or their image quality between those two groups (P >0.05);and there is a statistical significance in the ability of showing other levels of IPA and their im-age quality.Conclusion It is feasible that we use retrospective ECG-gated 256 slices spiral CT scanning technique to show the IPA. And it could improve the ability of showing the IPA branches as well as the image quality.

Objective To evaluate the effect of preoperative aescuven forte tablets in the prevention of procedure for prolapse and hemorrhoids(PPH) complications.Methods One hundred and seventy-nine patients underwent PPH in the Erlonglu Hospital of Beijing from June 2013 to June 2014 were divided into postoperative aescuven forte tablets group (n =90) and preoperative aescuven forte tablets group (n =89).The postoperative pain,bleeding,retention of urine,anal swelling,length of stay in hospital and recurrence were compared between the two groups.Results The incidence of postoperative pain,bleeding,retention of urine,anal swelling were lower than in the preoperative aescuven forte tablets group (pain after operations rate:7.9％ (7/89) vs.20.0％ (18/90),x2 =5.484,P＜ 0.05;Wound bleeding:0 vs.6.7 ％ (6/90),P =0.026;retention of urine:9.0％ (8/89) vs.20.0％(18/90),x2 =4.370,P＜0.05;anal swelling:1.1％(1/89) vs.8.9％(8/90),P=0.035),and length of stay in hospital was shorter(11.9±3.9) d vs.(13.3 ±5.0) d,t=3.134,P＜0.05).There was no recurrence both of two groups follow up for more than 2 months.Conclusion Preoperative oral aescuven forte tablets of 2 h can reduce PPH complications and worthy of popularization and application.

Objective To observe the role of brain-derived neurotrophic factor (BDNF) in the plasticity of enteric nervous-smooth muscle system,and to investigate the effects of BDNF induced plasticity on gastrointestinal motility in mice.Methods Male hybrid BDNF knockout (BDNF+/) mice and wild type (BDNF+/+) mice were selected,eight in each group.Gastrointestinal motility of BDNF+/+ mice and BDNF+/ mice were tested and compared.Longitudinal muscle strips of mice colon smooth muscle were prepared.The effects of carbachol (1 × 10 5 mol/L) and BDNF (1 × 10 7 mol/L) on contractile function of muscle strips were observed.And the effects of tetrodotoxin (TTX,1 × 10-6 mol/L) on BDNF induced contractile function of muscle strips were also studied.The changes of the density of mice intestinal myenteric plexus and the expression of smooth muscle α-actin (α-SMA) in colon smooth muscle were detected by immunohistochemical techniqne.The ultrastruetural alterations of myenteric plexus,neuromuscular junction (NMJ) and smooth muscle cells were detected by transmission electron microscope (TEM).T-test or Rank sum test was performed for comparison between groups.Results Number of feces particles and water content in feces of BDNF+/-mice ((3.80±0.75) and (39.60±1.47)％) were both lower than those of BDNF+/+ mice ((6.30± 1.03) and (51.00± 1.61) ％),and the differences were statistically significant (t=4.792,12.827;both P＜0.05).Carbachol (1 × 10-5 mol/L) could significantly increase contraction activity of smooth muscle of BDNF+/+ mice (R =3.26 ± 0.43) and BDNF+/-mice (R=2.15 ± 0.36),and the difference was statistically significant (t=15.754,9.632;both P＜0.05).The effects on contraction exciting of smooth muscle strips of BDNF+/+ mice were more significant than BDNF+/ mice,and the difference was statistically significant (t =5.972,P＜0.05).BDNF could significantly increase contraction of muscle strips of BDNF+/+ mice and R value increased from 1 to 1.41±0.09,and the differences were statistically significant (t=13.674,P＜0.05).TTX could obviously inhibit the excitatory effects of BDNF,R value decreased from 1.41 ± 0.09 to 1.03 ± 0.04 (t=11.692,P＜0.05).The density of myenteric plexus of BDNF+/ mice (median 5.8％,interquartile range 4.2％-7.0％) was significantly lower than that of BDNF+/+ mice (median 9.0％,interquartile range 7.1％-10.8％),and the difference was statistically significant (Z =3.730,P＜ 0.05).The expression of α-SMA of BDNF+/-mice (median 33.4％,interquartile range 28.8％-38.5％) was significantly lower than that of BDNF+/+ mice (median 44.6％,interquartile range 39.2％-48.8％),and the difference was statistically significant (Z=4.565,P＜0.05).The results of TEM indicated ultrastructural alterations of myenteric plexus,NMJ and smooth muscle in BDNF+/-mice.Conelusionss BDNF could induce the plasticity of morphology and function in enteric nervous-smooth muscle system,which may play an important role in mice gastrointestinal motility.