Objective To investigate the role of phosphatidyl-inositol 3-kinase-Akt (PI3k-Akt) signal pathway in the attenuation of ischemia-reperfusion (I/R) injury by sevoflurane preconditioning in isolated rat hearts. Methods Ninety-six adult male SD rats weighing 220-280 g were randomly divided into 6 groups ( n = 16 each): sham operation group (group S); I/R group; sevoflurane preconditioning group (group SP); wortmannin group (group W); dimethyl sulfoxide (DMSO) group (group D) and sevoflurane preconditioning + wortmannin group (group SW) . Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5%C02 at 37 ℃ . The hearts were continuously perfused for 180 min in group S. After 15 min of equilibration, the isolated hearts were subjected to 30 min of ischemia followed by 120 min of reperfusion in SP, W, D and SW groups. Croups SP, W, D and SW received 10 min of perfusion with K-H solution containing 2. 4% sevoflurane, 100 nmol/L wortmannin, 20 μmol/L DMSO, and 2.4% sevoflurane + 100 nmol/L wortmannin, respectively, followed by 5 min washout before I/R. Eight hearts in each group were selected and HR, left ventricular end-diabetic pressure (LVEDP), left ventricular developed pressure (LVDP), and ± dp/dtmax were recorded at the end of equilibration and at 15 min of reperfusion, Myocardial tissues were obtained at 15 min of reperfusion for determination of apoptosis (by TUNEL) and phosphorylated Akt (p-Akt) expression (by Western blot) . Another 8 hearts were selected at 120 min of reperfusion for determination of myocardial infarct size by TTC staining. Result Compared with group S, LVDP and ± dp/dt,^ were significantly decreased and LVEDP was significantly increased in groups I/R, SP, W, D and SW, and myocardial p-Akt expression was up-regulated in groups I/R, SP and D ( P ＜ 0.05). Compared with group I/R, LVDP and ± dp/dtmax were significantly increased, LVEDP and apoptosis index were significantly decreased, myocardial p-Akt expression was up-regulated, and myocardial infarct size was significantly reduced in group SP (P ＜0.05) . Conclusion Activation of PI3K-Akt signal pathway is involved in the attenuation of I/R injury by sevoflurane reconditioning in isolated rat hearts.

Objective To observe the levels of HIF-α and TGF-β in the liver tissue,change of serum transaminase in different phases after paraquat (PQ) toxicity and liver histopathology change in PQinduced liver toxicity of rats models in order to analyze the relationship between HIF-1 α and hepatic toxicity induced by PQ.Methods A total of 48 healthy SD rats were randomly (random number) assigned into 2 groups:PQ poisoning group (n =42,20％ PQ solution,50 mg/kg in 1mL) and control group (n =6,normal saline 1mL).Rats were separately sacrificed and liver tissue samples were harvested at 2,6,12,24,48,72 h and 120 h after PQ administered by gastric lavage.Arterial blood gas (ABG) analysis and serum alanine transaminase were assayed.Liver tissue HE and Masson staining and immunochemistry changes were also investigated.HIF-1α and TGF-β levels were detected by Western blot technique.Results Lactic acid level was significantly higher in PQ group than that in control group at 6 h after PQ exposure (P ＜ 0.05).The level of serum alanine transaminase was increased and significantly higher if PQ group than that in control group at 2 h after PQ poisoning (P ＜ 0.05).The level of HIF-1 α and TGF-β protein in the liver tissue were up-regulated significantly and higher in PQ group than those in control group at 2 h after PQ exposure (P ＜ 0.05).The hepatocytes necrosis and inflammatory cells infiltration were observed around portal area 2 h after PQ poisoning.At 12 h after PQ exposure,the periportal area filled with necrosis of hepatocytes and the necrosis began to extend,and huge amounts of inflammatory cell infiltration were found.Cord-like fibroplasia was found.All the histological changes were around central portal area and were enlarged gradually.Conclusions The results show that there are increased level of HIF-1 α in the early stage of PQ poisoning rats.The liver fibroplasia may be associated with increasing the level of TGF-β promoted by HIF-1α.

Aim To express hepatitis C virus glycoprotein E (gE) deleting carboxy-terminal 31 amino acids, and detect anti-E antibody in HCV patients using expressed gE. Methods E gene derived from HCV H strain was inserted into baculovirus transfer vector containing a polyhedrin promotor. The recombinant plasmid was cotransfected into insect cell sf9 with a viral expression vector. The expression of gE was analyzed with Western blot, and the cells were used for dectecting antibodies against E1 and E2 in 35 hepatitis C patients by indirect immunofluorescence. Results A series of proteins with different relative molecular masses(Mr) could be detected by Western blot. Results from indirect immunofluorescence staining showed and only 4 patients were anti-E antibody positive gE was located in cytoplasm. Conclusion HCV gE is expressed successfully in insect cells, the study lay the foundation for further developing HCV vaccine.

OBJECTIVETo explore the effect of peimine on excision repair cross-complementation 1 (ERCC1) mRNA and lung resistant protein (LRP) expressions in A549/cisplatin (DDP) multidrug resistance (MDR) cell line.

METHODSLung cancer A549/DDP cells were cultured in vitro.Cells at logarithmic growth phase were divided into 4 groups, i.e., the blank control group, the DDP group, the ligustrazine group (DDP+ligustrazine), the peimine group (DDP + peimine). After 48-h drug action, ERCC1 mRNA expression was detected by RT-PCR and LRP expression detected by cell immunofluorescence.

RESULTSThere was no statistical difference in expression levels of ERCC1 mRNA and LRP between the DDP group and the blank control group (P > 0.05). Compared with the DDP group, expression levels of ERCC1 mRNA and LRP obviously decreased in the ligustrazine group and the peimine group (P < 0.05). They were obviously lower in the peimine group than in the ligustrazine group (P < 0.05).

CONCLUSIONSPeimine could reverse MDR of A549/DDP cell line. Its mechanism might be associated with down-regulating ERCC1 mRNA and LRP expression levels.

Cell Line, Tumor ; Cevanes ; pharmacology ; Cisplatin ; DNA-Binding Proteins ; genetics ; Down-Regulation ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; drug effects ; Endonucleases ; genetics ; Humans ; Low Density Lipoprotein Receptor-Related Protein-1 ; genetics ; Lung Neoplasms ; RNA, Messenger ; metabolism

BACKGROUND: Research has been reported that serum soluble CD146 (sCD146) expression was improved on the surface of endothelial cells and activated T cells by the stimulation of inflammatory factor. Therefore, it predicts that CD146 may participate in inflammatory reaction of tissue.OBJECTIVE: To investigate the expression and clinical significance of serum sCD146 in peripheral blood from patients with ankylosing spondylitis.METHODS: A total of 62 patients with ankylosing spondylitis were selected from the Sixth People's Hospital AffiUated to Shanghai Jiao Tong University. All patients were divided into two groups: active group (n=46) and inactive group (n=16); while, 20 healthy subjects were selected as the control group. Indicators including Bath Ankylosing SpondyUtis Disease ActivityIndex (BASDAI),Bath Ankylosing Spondylitis Functional Index (BASFI), patient's global assessment (PGA), night pain, visual analogue scale (VAS),morning stiffness time, erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP) were measured in all patients. The serum concentration of sCD146 from 62 patients with ankylosing spondlitis and 20 healthy controls was measured by enzyme-linked immunosorbent assay. Westergren method was used to measure ESR and immunoturbidimetry for CRP. Clinical data of the patients were collected as well.RESULTS AND CONCLUSION: sCD146 levels of patients with ankylosing spondlitis were significantly higher than normal control group (P ＜ 0.05). The sCD146 expression in the active group was significantly higher than inactive and normal control groups (P ＜0.05). Positive correlations were observed between sCD146 and BASDAI index of patients with ankylosing spondlitis (P ＜ 0.05).The sCD146 levels of ankylosing spondUtis patients with peripheral joint involvement were significantly higher than the patients with axial involvement alone or the normal controls (P ＜ 0.05).The expression level of sCD146 in peripheral blood was positively correlated with disease activities of patients with ankylosing spondlitis. It may play important roles in the pathogenesis in ankylosing spondlitis.

Objective:To study the inhibitory effect of deoxyschizandrin on the growth of brain glioma C6 cells, and to explore its mechanism.Methods:The rat glioma C6 cells were cultured and divided into control group,50, 100,and 200 mg·L-1 deoxyschizandrin groups.The proliferation rates of C6 cells were examined by MTT assay;the changes of cell cycles were examined by flow cytometry;the expression levels of CyclinD1,Bax,Bcl-2 and Caspase-3 proteins in supernant were detected by ELISA assay. Results:Compared with control group, the proliferation rates at 24 and 48 h in 50,100,and 200 mg·L-1 deoxyschizandrin groups were significantly decreased (P <0.01),and the proliferation rates at 72 h in 100 and 200 mg·L-1 deoxyschizandrin groups were significantly decreased (P < 0.05 or P < 0.01 ). Compared with control group, the percentage of cells at SubG1 phase in 200 mg·L-1 deoxyschizandrin group was increased (P < 0.05 ), and the percentage of cells at S phase was decreased (P <0.05).Compared with control group,the expression levels of CyclinD1 in 100 and 200 mg· L-1 deoxyschizandrin groups were decreased (P < 0.01 );the expression levels of Bax protein in deoxyschizandrin groups were increased (P < 0.05 or P < 0.01 ), and the expression level of Bcl-2 protein in 200 mg · L-1 deoxyschizandrin group was decreased (P < 0.01 ), and the Bax/Bcl-2 value in deoxyschizandrin groups were increased (P < 0.01 ); the expression level of Caspase-3 protein in 200 mg · L-1 deoxyschizandrin group was increased (P < 0.01 ).Conclusion:Deoxyschizandrin could inhibit the growth of glioma cells through down-regulating the expression levels of CyclinD1 protein and up-regulating the expression levels apoptotic factors Bax and Bcl-2.

Objectives To evaluate pediatric neck masses with magnetic resonance imaging (MRI). Methods In this retrospective study, 140 children with neck masses underwent MRI were collected from May 2006 to December 2013. Of them 34 cases went through pathological examinations. The results of MRI diagnosis and pathology were compared in 34 cases. Results In 140 children with neck masses diagnosed by MRI, 103 (73.6%) cases were benign lesions, including 62 vascular malformations, 30 hemangiomas, then cysts, hamartoma, infectious lumps etc., 29 (20.7%) were malignant tumors, including 22 lymphomas, 3 rhabdomyosarcomas, 3 Langerhans cell histiocytosis, 1 neuroblastoma, and 8 (5.7%) cases were undeter-mined masses. Four in 103 cases with benign lesions were performed by pathological examination and all had been con-firmed. Tewenty-five in 29 cases with malignant tumors were performed by pathological examination and 22 cases had been confirmed. Conclusion MRI can help to diagnose the pediatric neck masses and to guide the treatment and follow-up.

Objective To assess the diagnostic potential of circulating galactomannan (GM),(1,3)-β-D-glucan (BG), and a combination of both biomarkers among high-risk pediatric patients.Methods Circulating GM antigen was detected by ELISA kits (Platelia~(TM) Aspergillus) and BG antigen by a turbidimetric kinetic method (GKT-5M Set Kinetic Fungus Detection Kit).Positive tests were defined by two consecutive values of GM index ≥0.5 or by a single value ≥0.8, and by BG detection ≥ 10 pg/ml.Results A total of 130 patients were enrolled.Two was identified with proven IFI, twenty probable IFI.Sensitivity, specificity were 81.8%, 82.4% for plasma BG detection, respectively; 75.0%, 94.4% for GM detection, respectively; 50.0%, 96.3% for combined GM and BG detection, respectively.Conclusions Both circulating GM and BG detections are available for most of the common pathogens and demonstrated desirable sensitivity and specificity among pediatric high-risk population.Both assays can be used as prospective screening tools.Combination of detections of both biomarkers would improve specificity for IA diagnosis.

Objective To investigate the effect of specific cGMP-dependent protein kinase G (PKG) inhibitor (D)-DT-2 on the contractile function of rat vascular rings after being exposed to lipopolysaccharide (LPS).Methods The experiment was performed in 2 parts.Part Ⅰ:The Sprague-Dawley rat thoracic aortic rings were randomly divided into 3 groups (n =5 each):KH group,DT-2 group and (D)-DT-2 group.KH,DT-2 and (D)-DT-2 were added to the aortic ring after being dilated with 8-Br-cGMP 50 μmol/L for 25 min and the changes in tension of vascular rings were measured.Part Ⅱ:The rat thoracic aortic rings were randomly divided into 4 groups (n =5 each):control group,LPS group,LPS-DT2 group and LPS-(D)-DT2 group.After being incubated with LPS for 3 h in vitro.The Emax and EC50 were compared among the 4 groups.Results Part Ⅰ:Both DT-2 and (D)-DT-2 could contract the vascular rings dilated with 8-Br-cGMP and the Emax was significantly higher in (D)-DT-2 group than DT-2 group (P ＜0.05).Part Ⅱ]:Both DT-2 and (D)-DT-2 significantly improved the contractile function of vascular ring after being exposed to LPS.Emax was significantly higher,while EC50 was lower in groups DT-2 and (D)-DT-2 than in LPS group (P ＜0.01).Emax was significantly increased,while EC50 was decreased in LPS-(D)-DT-2 group as compared with LPS-DT-2 group (P ＜ 0.05).Conclusion PKG inhibitor can improve the contractile function of the vascular rings incubated with LPS and the efficacy of (D)-DT-2 is better than DT-2 in recovering the vascular reactivity.

OBJECTIVETo observe the clinical efficacy of TCM for reinforcing Shen and activating blood circulation to dredge Channels in auxiliary treating chronic aplastic anemia (CAA), and to explore its mechanism.

METHODSSeventy-nine patients with CAA were randomly divided into the treated group treated with TCM plus western medicine and the control group treated with western medicine alone. The clinical efficacy was observed and levels of plasma sFas and sFasL before and after treatment were determined by ELISA.

RESULTSThe basic cure rate and total effective rate in the treated group were 37.50% and 87.50%; while in the control group were 20. 51% and 61.54% respectively. Comparison between the two groups showed significant difference (P < 0.01). The sFas level was markedly lower in both groups before treatment, but after treatment, it got elevated in the treated group significantly (P < 0.05), as compared with that in the control group, the difference was significant (P < 0.01).

CONCLUSIONTCM is effective in treating CAA with less toxic and side-effect, the mechanism might be the inhibition on over apoptosis of hematopoietic cells in CAA patients through regulating immune function of organism.

Adolescent ; Adult ; Aged ; Anemia, Aplastic ; drug therapy ; Apoptosis ; drug effects ; Child ; Chronic Disease ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hematopoietic Stem Cells ; pathology ; Humans ; Male ; Middle Aged ; Phytotherapy ; Single-Blind Method ; Ubiquinone ; therapeutic use