Objective·To investigate the effects of insulin-like growth factor 1 on prion encoding gene (PRNP) expression and amyloid precursor protein (APP) metabolism of PC12 cells. Methods·After PC12 cells were treated with 20, 40, 80 ng/mL IGF-1 for 24 h, real-time PCR was used to detect the mRNA expression levels of PRNP, and Western blotting was used to detect the protein levels of AKT, phosphorylation of AKT (pAKT), ERK and phosphorylation of AKT (pERK). The level of β-amyloid 42 (Aβ42) in supernatant fluid was detected by ELISA. Results·Compared with the blank control group, the expression of PRNP mRNA in Alzheimer's disease (AD) model group was increased significantly (P<0.01). The expression of PRNP mRNA was significantly increased after cells were treated with 40 and 80 ng/mL IGF-1 for 24 h (P<0.01). There was no significant difference in APP protein expression among AD model group and three IGF-1 treatment groups (P>0.05). Compared with the blank control group, the level of Aβ42 in supernatant fluid of model group was decreased significantly with the increasing of IGF-1 concentration. The expression level of Aβ42 was decreased significantly in 40 and 80 ng/ml IGF-1 treatment group (P<0.05). The expression of AKT/pAKT, ERK/pERK in AD model group was significantly increased along with the increasing of IGF-1 concentration (P<0.05). Conclusion·IGF-1 could regulate the expression of PRNP gene and effect the metabolism of APP, which may be associated with PI3K/AKT, MAPK/ERK1/2 signaling pathway.

Objective To observe the effects of indole-3-carbinol on neointimal hyperplasia and restenosis of rat artery after balloon injury and the possible mechanisms.Methods Balloon dilation was used to establish the neointimal injury model of left carotid artery in rats.Twenty Sprague-Dawley rats were randomly divided into single balloon dilation group(control group)and balloon dilation followed by indole-3-carbinol therapy group(therapeutic group).After balloon dilation,indole-3-carbinol(12.5,25,50 mg/d)was applied to the rats for 7 days respectively.The rats were killed two weeks after balloon dilation and the injured vascular specimens were harvested for pathologic examination and immunohistochemical staining.Results ①The neointimal thickness,neointimal area in the therapeutic groups were significantly less than that of the control group(P

0.05).The intimal thickness was decreased in high-dose DIM group compared with model group and low-dose DIM group(P

This study is to investigate the effect of artesunate on transforming growth factor-beta1 (TGF-beta1) induced epithelial-mesenchymal transition (EMT) and its possible mechanism. After the in vitro cultured RLE-6TN cells were treated with TGF-beta1 then artesunate intervened on it, after 24 h, expression of the markers of mesenchymal cell was assayed using Western blotting and real-time PCR analysis. Western blotting was also used to detect the effect of TGF-beta1 on the Smad3 and Smad7 expressions of RLE-6TN cells. Morphological alterations were examined by phase-contrast microscope, and ultrastructure changes by electron microscope. Incubation of RLE-6TN cells with TGF-beta1 resulted in the up-regulation of the expression of the mesenchymal cell markers, after artesunate intervened on it, resulted in the down-regulation of the expression. Meanwhile, incubation with artesunate intervened on RLE-6TN cells could lead to the apparent down-regulation of the expression of Smad3 and up-regulation of Samd7 and the transition of RLE-6TN cells to mesenchymal-like by TGF-beta1 induction, after artesunate intervened on it, RLE-6TN cells to epithelial-like. TGF-beta1 induced epithelial-mesenchymal transition process; artesunate can inhibit TGF-beta1-induced epithelial-mesenchymal transition process, the possible mechanism is up-regulation of the expression of Smad7 and down-regulation of the expression of Smad3, meanwhile inhibits phosphorylation of Smad3.

Objective To explore the prognostic value for cardiovascular events of elevated NT-proBNP in coronary heart disease(CHD) patients with metabolic syndrome(MS).Methods The level of NT-proBNP was measured in 165 CHD patients with MS diagnosed by coronary angiography.After 2 years follow-up,we recorded the corresponding cardiovascular events in those patients.Results The NT-proBNP level was much higher in ACS group than that in SCHD group( P ＜ 0.01 ).There were totally 45 events after 2 years.After adjustment of traditional risk factors including hypertension,diabetes,smoking history and high sensitivity C reactive protein( hs-CRP),Cox proportional hazard analysis indicated that high NT-proBNP level( ＞ 111.58ng/L)was an independent risk factor for future events (relative risk 2.67,P ＜0.01 ).Conclusion Elevated NT-proBNP level might be one of the independent risk factor for cardiovascular events in CHD patiens with MS.

Objective To study the function of the toll-like receptor-4 (TLR-4) signaling pathway in the synthesis and secretion of pulmonary artery smooth muscle cells of rats with COPD. Methods The primary pulmonary artery smooth muscle cells (PASMCs) of rats with COPD were digested, separated and purified. Then they were randomly divided into control group, LPS group, TLR4 inhibitor group (TAK242) and LPS + TLR4 inhibitor group. RT-PCR, Western blot were used to detect the expression level of TLR-4 and NF-κB among groups. The levels of IFN-γ and PDGF-AB in supernatant with PASMCs in each group were detected by ELISA. Results LPS increased the expression of TLR-4、 NF-κB and the levels of IFN-γ and PDGF-AB. The expression of TLR4, NF-κB and the levels of IFN-γ and PDGF-AB were significantly reduced after inhibiting the expression of TLR4(P < 0.05). Conclusion TLR-4 signaling pathway involved in the inflammatory response and pulmonary vascular remodeling which increased the synthesis and secretion of IFN-γ and PDGF-AB in PASMCs. It provides a theoretical approach for the early intervention of clinical with COPD.

ObjectiveTo study the characteristic of apoptosis and the expression of inducible nitric oxide synthase(iNOS),as well as relation between them after spinal cord injury (SCI) in rats.Methods84 adult SD rats were randomly divided into three groups,and made into mild,moderate and severe acute SCI models.The rats were killed at 4h,8h,1d,3d,7d,14d and 21d after surgery.The histopathology changes in spinal cord tissue were observed with HE staining microscopic examination.The expression of iNOS was determined by immunohistochemistry and the apoptosis was labeled by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL).ResultsThe histopathology changes in spinal cord tissue deteriorated with increasing in injury degree.A little iNOS immunoreactivity (iNOS-IR) was detected in nerve cells,gliocytes,ependymocytes and blood vessel endotheliocytes of normal and vertebra shelf operated controls,increasing at 8 h, and in flood tide in 7 d. The expression of Bax and the rates of TUNEL positive cells were similar with that of iNOS after SCI. There was positive correlation between expression of iNOS and degree of primary SCI ,apoptosis index(r=0.414,P<0.01;r=0.854,P<0.01).Conclusions Expression of iNOS and Bax increase and a large number of TUNEL positive cells present after SCI in rats. There is positive correlation between expression of iNOS and degree of primary SCI,apoptosis index.

Objective To observe the changes in hypersplenism after orthotopic liver transplantation(OLT) and investigate the effect of OLT on hypersplenism. Methods Based on the clinical data of 14 eligible OLT patients operated on in our hospital during two and a half years, an analysis of the pre operative values of the thickness of the spleen, blood WBC and PLT count was made,and the postoperative pattern of changes of portal flow velocity was observed. Results Blood WBC and PLT count returned to normal 1 month after the operation, and the thickness of spleen reduced about 17.0% in the first month , but had no additional change at 1 year later. Portal flow velocity increased significantly 1 month after operation , then decreased slowly in the first year. Portal flow velocity , blood WBC and PLT count were all significantly related to the thickness of the spleen. Conclusions The high velocity of portal flow after OLT was mostly attributed to increased flow from the splenic vein; the main cause of the decrease in the level of blood WBC and PLT in hypersplenism before operation is augmentation of splenic volume; the recuperation of hypersplenism after OLT relies on the extent of reversion of splenic volume. It is not necessary to perform splenectomy in patients with hypersplenism when they receive OLT.

Objective To evaluate the effects of different selection criteria on the prognosis of hepatocellular carcinoma(HCC)patients undergoing liver transplantation(LT)and to evaluate a new criterion.Methods A retrospective analysis was performed on 81 consecutive patients with HCC who underwent LT.The survival rates of the patients who met different criteria such as Milan.UCSF(University of California San Francisco UCSF).and Pittsburgh(Pitt)modified TNM criteria were calculated by KaplanMeier method,and the value of different criteria was evaluated.The Long-Rank test and COX proportional hazards regression model were performed to analyze the prognostic factors.the model of criteria was established according the most important prognostic factors.Using the Kaplan-Meier method,the suitable cut-offs of every variable ifl the model were found by comparing the survival and the number of the patients who met the cut-off,and considering the significant difference between the patients who met and exceed the cut-off at the same time.Resuits The 1,2,3-year accumulative survival rates of the 19 patients who met Milan criteria were 87.7%,87.7%,and 52.6%respectively:the 1,2,3-year disease free survival rates of them were 88.9%,72.7%,and 72.7%respectively.The 1,2.3-year accumulative survival rates of the 26 patients who met UCSF criteria were 87.2%,80.5%,and 55.2%respectively;the 1,2,3-year disease free survival rates of them were 84.1%,68.4%,and 68.4%respectively.With our new expanded criterion as of solitary tumor≤8 cm in diameter.or no more than 3 tumors,with the largest≤6 am,and a total tumor diameter≤10 cm.there was no significant difierence in 1,2,3-year sunrival rates and disease free survival rates(89.0%,81.8%,71.8%,and 81.9%,72.4%,72.4%.)as compared with Milan or UCSF criteria.but with this new criterion more patients(a=41)would be eligible for transplantation with a comparable long term survival.and the difference of the accumulative survival rates and disease free survival rates of the patients who met and exceed the new criteria was significant(P＜0.05).Conclusion The new indication is acceptable because the criteria does not adversely impact survival.

[Abstract ] Objective Chronic obstructive pulmonary disease (COPD) is closely related to pulmonary hypertension .This study was to investigate the correlation of the expression of high mobility group protein B 1 ( HMGB1) with its synthesis and secretion of interferon-γ( IFN-γ) in the distal pulmonary arterial smooth muscle cells ( PASMCs ) of COPD rats and its clinical significance . Methods We established COPD models in rats , primarily cultured and identified PASMCs , and detected the synthesis and secretion of cytokine in the PASMCs induced by cigarette smoke extract (CSE) and/or lipopolysaccharide (LPS).The PASMCs in the control, CSE, LPS, and CSE+LPS groups were cultured with anti-HMGB1 antibodies at 0.2 mL 1∶1000.At 12, 24, 48 and 72 hours of inter-vention, the expression of HMGB1 in the PASMCs was detected by Western blot and the concentrations of HMGB 1 and IFN-γin the cell culture supernatant were measured by ELISA , followed by a correlation analysis . Results Light microscopy manifested a peak -valley growth pattern or fusiform shape of the cells , which were identified as PASMCs by α-actin immunohistochemistry and immuno-fluorescence .Western blot and ELISA showed statistically significant differences in the expression of HMGB 1 and concentrations of cell supernatant HMGB1 and IFN-γat 12 hours between any two of the four groups (P<0.05).The HMGB1 expression level was positive-ly correlated with the concentration of IFN-γ(r=0.91), obviously decreased after 12 hours of intervention with anti-HMGB1 antibod-ies as compared with the corresponding experimental groups ( P <0.05).Western blot and ELISA showed no significant differences within the same group at 12, 24, 48 and 72 hours (P>0.05). Conclusion The expression level of HMGB1 was positively correla-ted with the synthesis and secretion of IFN-γin the distal PASMCs of the COPD model rats , and anti-HMGB1 antibodies provide a new in-tervention target for the clinical treatment of COPD and inhibition of its systemic inflammatory response .