Objective To investigate the expression of scaffold protein palladin in pancreatic ductal adenocarcinoma (PDAC) tissues and to discuss its clinicopathological significance.Methods 56 samples of PDAC and corresponding adjacent normal pancreas (NP) tissues were collected.Another 10 samples of chronic pancreatitis (CP) tissues were collected.Western blot analysis and immunohistochemistry assay were performed to detect the expression of protein palladin.The correlation of palladin expression with clinicopathological factors of PDAC was analyzed.Results Western blot analysis revealed that the expression of palladin in PDAC,CP and NP tissues were respectively 0.93±0.07,0.41±0.07 and 0.20±0.06,and the expression of palladin was significantly increased in PDAC tissues compared with NP and CP tissues (P ＜ 0.05),and its expression was significantly increased in CP tissues compared with NP tissues (P ＜ 0.05).Immunohistochemical staining showed that palladin was mainly expressed in activated myofibroblasts in PDAC tissues.The rate of palladin expression was 79 ％ (44/56),which was higher than that in NP tissues (2/10) and CP tissues (4/10),and its expression was found to be correlated with the degree of tumor differentiation,lymph node metastasis and clinical TNM classification (P ＜ 0.05),and it had no correlation with patient' s sex,age,tumor location and distant metastasis (P ＞ 0.05).Conclusions Scaffold protein palladin is highly expressed in PDAC tissues,and it is expressed in the activated myofibroblasts within tumor microenvironment.Scaffold protein palladin may be involved in the invasion and metastasis of PDAC.

Objective To evaluate diagnostic value and clinical significance of procalcitonin (PCT)in infection in intensive care unit (ICU)patients.Methods 96 ICU patients in a hospital between September 2011 and March 2012 were selected for study,levels of patients’PCT,high-sensitivity C-reactive protein (HsCRP)and white blood cell (WBC)count were detected,statistical analysis were conducted.Results Compared with non-bacteria infected patients,serum PCT and HsCRP levels in all bacteria infected patients increased,the difference were significant (Z=-6.102;-3.918,both P <0.05 );WBC count was not significantly different(Z =0.212.P >0.05).PCT sensi-tivity,specificity,positive predictive value,and negative predictive value for diagnosing infection was 82.35%, 67.86%,86.15%,and 61 .29% respectively;receiver operating characteristic (ROC)curve of PCT,HsCRP,and WBC was 0.898,0.755,and 0.581 respectively.Conclusion There are higher sensitivity and specificity of PCT to predict infection,which is helpful for early detection of infection in critically ill patients.

AIM:As a factor that can improve cell growth,there are few studies about the effect of insulin-like growth factor Ⅰ(IGF-Ⅰ) on the apoptosis of endothelial cell.The study investigated the inhibition and mechanism of IGF-Ⅰ on the apoptosis of human umbilical vein endothelial cells(HUVEC) induced by oxidized low density lipoprotein(ox-LDL).METHODS:The experiment was performed in the Institute of Cardiovascular Disease,Union Hospital of Huazhong University of Science and Technology from December 2006 to July 2007.①Fresh human umbilical cord was obtained(the informed consent) to isolate and culture HUVECs.The cells were divided into four groups.Except the control group,HUVEC cells were cultured with IGF-Ⅰ(1?10-9mmol/L),ox-LDL(200 mg/L)+IGF-Ⅰ(1?10-9mmol/L),and ox-LDL(200 mg/L),respectively after cultured for 24 hours.②Cell viability was determined by MTT assay,morphology and apoptosis by DAPI fluorescence staining,and expressions of caspase-3 were analyzed.RESULTS:①Ox-LDL could significantly inhibit HUVEC cell proliferation.After treated with both IGF-Ⅰand ox-LDL,the cell proliferation increased obviously compared with the cells treated with ox-LDL(P

This study examined the change of p16(INK4a) and PNCA protein expression in myocardium after injection of hIGF-1 gene modified skeletal myoblasts into post-infarction rats. HIGF-1 gene modified skeletal myoblasts (hIGF-1-myoblasts) were injected into hind limb muscles of 18 post-infraction rats (experimental group). Primary-myoblasts were injected into 18 post-infraction rats (control group) and 12 non-infarction rats (sham group). Expression of p16(INK4a) and PCNA protein in myocardiums were separately detected immunocytochemically 1, 2 and 4 weeks after the inuection. The level of hIGF-1 and rIGF-1 protein in serum and myocardium were detected by enzyme-linked immunosorbent assay (ELISA). Compared with the sham group, the percentage of p16(INK4a) and PCNA positive cells reached a peak after 1 week in the control group and the experimental group (P<0.01). Moreover, the percentage of p16(INK4a)-positive cells in the experimental group was lower than in control group whereas the percentage of PCNA-positive cells was lower in the control group than in the experimental group (P<0.01). The percentage of p16(INK4a)-positive cells in the experimental group and the percentage of PCNA-positive cells in the control group were close to that in the sham group from the 2nd week (P>0.05). ELISA analysis disclosed that the myocardium level of rIGF-1 protein increased gradually in the controls and especially in the experimental group (P<0.01). The serum level of rIGF-1 decreased significantly in post-infraction rats, but these conditions were improved in the experimental group (P<0.01). The hIGF-1 protein in serum and myocardium were detected from the 1st week to the 4th week in the experimental group. Statistical analysis revealed significant associations of myocardium level of hIGF-1 protein with expression of p16(INK4a) and PCNA protein (r=-0.323, P<0.05; r=0.647, P<0.01). It is concluded that genetically hIGF-1-myoblast provides a means for constant synthesis and release of hIGF-1. It could not only improve the expression of rIGF-1 and PCNA protein in myocardium, but also suppress the expression of p16(INK4a) protein for 30 days in post-infraction rats. Myoblasts-mediated IGF-1 gene therapy may provide a new alternative for the clinical treatment of heart failure.

ObjectiveTo explore the effects of recombinant human growth hormone(rhGH)on myocardial inflammatory cytokine expression and heart function in rats with acute myocardial infarction (AMI). MethodsRats with AMI induced by left anterior descending coronary branch ligation were randomized to rhGH and control groups compared with sham-operated group. The effects of 4 weeks of therapy with GH starting 24 hours after myocardial infarction on myocardial cytokines expression and heart function were studied. Myocardial inflammation was examined by analyzing the myocardial cytokine production including the pro-inflammatory cytokines: interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α and the anti-inflammatory cytokine: IL-10. Echocardiography was used to evaluate heart function. ResultsThe levels of TNF-α, IL-1β, IL-6 and IL-10 in the infarcted and non-infarcted region of control group were markedly elevated compared to sham-operated group (all P＜0.05). After 4 weeks therapy, rhGH reduced the expression of TNF -α, IL-1β, IL-6 and increased IL-10 expression in the infarcted and non-infarcted region of rhGH group compared to control group (all P＜0. 05 ). Echocardiography showed that rhGH markedly improved left heart function (P＜0. 05 ). ConclusionsEarly rhGH treatment can improve heart function and myocardial inflammatory cytokine expression after AMI. One of immunopharmacologic mechanisms underlying the beneficial effects of rhGH on heart function improvement may involve the attenuation of pro-inflammatory cytokines and the increase of anti-inflammatory cytokine levels in cardiac myocytes.

Objective To investigate whether astragaloside Ⅳ can regulate the multidrug resistance of HepG2/GCS-resistant cell lines,restore the sensitivity of drag-resistant cell lines to adriamycin (ADM) and its mechanism.Methods We used recombinant GCS (shRNAS) and control recombinant plasmids and did the transfection with HepG2 cells.RT-PCR and Western blot were used to analyze the expression of GCS.Astragaloside Ⅳ cytotoxicity experiments and ADM were performed in experimental and control groups.Hoechst 33258 was detected in two groups,apoptosis was detected by flow cytometry,and protein expression of caspase 9,3,Bax,and bcl-2 were detected by Western blot.Results RT-PCR and fluorescence observation showed that GCS was highly expressed after the transfection.Western blot showed that compared with control group,and HepG2GCS group,GCS and MDR1 expression were higher than;Astragaloside Ⅳ cytotoxicity experiment showed tumor proliferation was not regulated by GCS(FHepG2GCS=0.308,FHepG2EV =0.216,FHepG2 =0.153,P＞ 0.05),ADM in vitro tumor cell inhibition experiments showed that HepG2GCS cells were resistant to ADM (50％ cell transplantation concentration were 7.5,7.5,15 μg/ml);Hoechst 33258 and flow cytometry showed that Astragaloside Ⅳ can restore ADM tumor inhibition;Western blot showed that compared to untreated HepG2EV and HepG2GCS cells,protein level of caspase 9,caspase 3 were increased in Ast+HepG2EV and Ast+HepG2GCS groups (t=7.17,P＜0.05).At the same time,Bax and Bcl-2 were significantly different in each group (P＜ 0.05).Conclusions Astragaloside Ⅳ reverses multidrug resistance in HepG2/GCS cell lines,restores its sensitivity to ADM,promotes apoptosis in tumor cells through the caspase pathway and Bax pathway,and thus plays an important role in cancer chemotherapy.

This study examined the change of p 16INK4a and PNCA protein expression in myocardium after injection of hIGF-1 gene modified skeletal myoblasts into post-infarction rats. HIGF-1 gene modified skeletal myoblasts (hIGF-1-myoblasts) were injected into hind limb muscles of 18post-infraction rats (experimental group). Primary-myoblasts were injected into 18 post-infraction rats (control group) and 12 non-infarction rats (sham group). Expression of p16INK4a and PCNA protein in myocardiums were separately detected immunocytochemically 1, 2 and 4 weeks after the inuection. The level of hIGF-1 and rIGF-1 protein in serum and myocardium were detected by enzyme-linked immunosorbent assay (ELISA). Compared with the sham group, the percentage of p16INK4a and PCNA positive cells reached a peak after 1 week in the control group and the experimental group (P＜0.01). Moreover, the percentage of p16INK4a-positive cells in the experimental group was lower than in control group whereas the percentage of PCNA-positive cells was lower in the control group than in the experimental group (P＜0.01). The percentage of p16INK4a-positive cells in the experimental group and the percentage of PCNA-positive cells in the control group were close to that in the sham group from the 2nd week (P＞0.05). ELISA analysis disclosed that the myocardium level of rIGF-1 protein increased gradually in the controls and especially in the experimental group (P＜0.01). The serum level of rIGF-1 decreased significantly in post-infraction rats, but these conditions were improved in the experimental group (P＜0.01). The hIGF-1 protein in serum and myocardium were detected from the 1st week to the 4th week in the experimental group. Statistical analysis revealed significant associations of myocardium level of hIGF-1 protein with expression of p16INK4a and PCNA protein (r=-0.323, P＜0.05; r=0.647, P＜0.01). It is concluded that genetically hIGF-1-myoblast provides a means for constant synthesis and release of hIGF-1. It could not only improve the expression of rIGF-1 and PCNA protein in myocardium, but also suppress the expression of p16INK4a protein for 30 days in post-infraction rats. Myoblasts-mediated IGF-1 gene therapy may provide a new alternative for the clinical treatment of heart failure.