Objective To investigate the effects of peroxisome proliferator-activated receptor-? (PPAR?) agonist Pioglitazone on treating the vascular dementia (VD) model.Methods SD rats were randomly divided into the sham-operated group,VD group,Pioglitazone high-dose group [20 mg/(kg?d)] and low-dose group [5 mg/(kg?d)].The VD models were made by modified Pulsinelli four-vessel occlusion method,and each group received corresponding treatment for 7 d.Then,Morris water maze test was applied to examine the place navigation escape latency.The number of PPAR? positive neurons and expression of PPAR? in the cerebral tissue were detected by immunohistochemistry.Results Compared to the VD group,the place navigation escape latencies in the two Pioglitazone treated groups were significantly shorter(all P

Objective To compare the therapeutic effects of different doses of pioglitazone, a kind of peroxisome proliferator-activated receptor γ (PPARγ) agonist, on vascular dementia and explore how pioglitazone affects cerebral ischemia. Methods Modified Pulsinelli's vessel ligation was used to establish a vascular dementia model in rats. Recognition, learning and memory were evaluated by Morris's water maze test. Immunoenzyme staining was used to determine the number of nerve cells. Immunofluorescence double-staining was used to examine the expression of PPARγ/nerve cells and PPARγ/astrocytes in different groups. Results Both in pioglitazone groups and sham-operation group, the latency was reduced significantly compared to that in control group (P<0.01). Sham-operation group had the largest number of neurons in the cortex, followed by low-dose pioglitazone group and high-dose pioglitazone group, and control group came last. Compared with control group, pioglitazone groups had more PPARγ expression in nerve cells, and the fluorescence intensity of PPARγ was stronger. Conclusion Pioglitazone can induce the expression of PPARγ in neuron endochylema and astrocyte endochylema to protect nerve cells, and then to improve spatial learning and memory function in VD rats.

Objective:This study aimed to investigate the effect of human lymphatic endothelial cells (HLECs) on proteins secreted by epithelial ovarian cancer (EOC) cells SKOV3-pm4 with highly directional lymphatic metastasis. Methods:The supernatants of the four groups of cultured cells (A, SKOV3;B, SKOV3+HLEC;C, SKOV3-PM4;and D, SKOV3-PM4+HLEC) were collected. The proteins of these cells were detected by antibody arrays and iTRAQ-2D-LC-MALDI-TOF/TOF/MS. The screened significantly differential proteins were further analyzed by bioinformatics and validated in the human serum and cell culture medium by ELISA. Results:Progranulin (GRN) and vascular endothelial growth factor A (VEGF-A) were upregulated between groups C and A. In addition, insulin-like growth factor binding protein-7 (IGFBP-7) and secreted protein acid rich in cysteine (SPARC) were downregulated between groups D and C. Comprehensive bioinformatics analysis revealed that IGFBP7 interacted with VEGFA. VEGF exhibited the highest expression in ovarian cancer and IGFBP7 exhibited the lowest expression compared with the serum of the normal control group. Statistically significant differences were observed between the two substances. Conclusion:The HLEC microenvironment is closely associated with directional metastasis in lymph nodes with differential proteins, including matricellular proteins and adhesion factors. In particular, the upregulation of VEGFA and GRN and the downregulation of SPARC and IGFBP7 were closely associated with the directional metastasis of EOC cells in lymph nodes.

ObjectiveTo explore the effect of blocking the JAK/STAT signal pathway on the activity of Caspase-3 in the synovial tissue of rheumatoid arthritis rats.MethodsFifty rat models of collageninduced arthritis,which had arthritis index more than 2 were divided into the model group,the low dosage of AG490 group,the medium dosage of AG490 group and high dosage of AG490 group.Inaddition,6 rats were treated as normal control group.Normal saline,AG490 1,5,10 mg·kg-1 ·d-1 were given by intraperitoneal injection.Then the volume claws and pathologic scores of the rat models were recorded and the activity of Caspase-3 in the synovial tissue were compared.The results were analyzed by one-way ANOVA and LSD-t or Tamhane's T2 test.ResultsThe arthritis of the CIA models progressed fast,the volume of the claws and the pathological score of them were significantly higher than those of the control group.At the same time,no Caspase-3 positive express could be detected in the control group,whilethe model group had slightly increased expression.After different dosages of AG490 were applied,the swollen of joints was significantly improved compared with the model group.The histopathological score of the medium AG490 dosage of group and high dosage group(2.7±0.8,1.8+0.9) were remarkably decreased than those of the model group(4.3+1.2),the differences were statistically significant (P＜0.01).In addition,the Caspase-3 expression in the low,medium and high AG490 dosage group ( 1.90±0.15,3.13±0.33,3.56+0.34) was significantly higher than that of the model group(1.48±0.18)(P＜0.05 or P＜0.01 ).ConclusionBlocking JAK/STAT signal pathway can increase the activity of Caspase-3,reduce the excessive proliferation of synovial tissue,and improve arthritis symptoms.

Objective To explore the correlation of bone metabolism levels and risk of coronary heart disease in elder women patients .Methods A total of 163 elder women patients were divided into three group:CON group ,CAD group ,and CHD group .We explored related atherosclerosis risk factors and factors related to bone metabolism .Results Compared with CON group ,there was no statistical significance in CAD group in factors related to bone metabolism(P>0 .05) .In CHD group ,serum 25‐OH‐Vitamin D significantly decreased and β‐C‐terminal telopeptide of type Ⅰ collagen significantly increased compared with CON group(P<0 .05) .Compared with CAD group ,the serum 25‐OH‐Vitamin C also significantly decreased andβ‐C‐terminal telopeptide of type Ⅰcollagen significantly increased(P< 0 .05) .Spearman correlation analysis showed that BMI ,HDL‐C ,triglycerides ,LDL‐C ,blood glucose and 25‐OH‐Vitamin D were correlated with coronary heart disease .With coronary heart disease as the dependent variable , the results showed lower LDL‐C ,25‐OH‐Vitamin D had independent predictive value for the risk coronary heart disease .Conclusion Lower 25‐OH‐Vitamin D levels in elder patients were positively correlated with coronary heart disease ,and it might also be an in‐dependent predictor .

Objective To establish the condition cultrue cell system and co-culture cell system with SKOV3/PM4,HUVEC and to study the changes of their biological characteristics. Methods The cells of SKOV3/PM4 and HUVEC were labeled with green and red fluorescent respectively. The cell supernatant of SKOV3/PM4 and HUVEC were collected respectively as the condition medium(e.g:the cell supernatant of HUVEC cells was used as SKOV3/PM4 condition medium)and to establish the condition cultrue cell system and the co-culture cell system of the two cell lines. In the condition cultrue cell system, The morphological changes of cells were observed by HE staining to calculate the mitotic index. The ultrastructural changes of the two cells were observed by transmission electron microscopy(TEM). The growth curve of the cells was determined by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry was used to analyzed the cell cycles.In the co-culture cell system, the interaction of the two cells were detected by laser scanning confocal microscope(LSCM). The expression of matrix metalloproteinase-2(MMP-2) and matrix metalloproteinase-9 (MMP-9) were detected by gelatin zymography. Results Compared with the single culture SKOV3/PM4, the cells which cultured in HUVEC condition medium showed the increase of pseudopodia and nuclear division,the mitotic index respectively were [(4.8 ± 0.8)%,(11.2 ± 0.3)%;P<0.05]. The growth rate was significantly increased. In cell cycles, it showed the declined cell ratio of G0/G1 phase, respectively[(69.4±3.6)%, (48.4±4.6)%;P<0.05] and the raised cell ratio of G2/M phase, respectively [(5.2±1.6)%, (24.9±2.2)%;P<0.05]. Compared with the single culture HUVEC,the cells which cultured in SKOV3/PM4 condition medium showed the significant morphological change and vacuolization in the cytoplasm, Nuclear division was increased and the mitotic index respectively were [(2.7±0.5)%, (5.7±0.6)%;P<0.05]. The growth rate was slightly declined. In cell cycles, it showed the raised cell ratio in G0/G1 phase, respectively [(51.4 ± 2.2)%,(79.0 ± 4.1)%;P<0.05] and the declined cell ratio in G2/M phase, respectively [(19.1±1.2)%, (3.3±0.5)%;P<0.05]. After co-culture for 48 hours, spontaneous fusion between SKOV3/PM4 and HUVEC cell was observed by the laser confocal microscope. Gelatin zymography assay showed that MMP-2 was not expressed in HUVEC cells, low-expressed in SKOV3/PM4 cells and high-expressed in the co-culture SKOV3/PM4+HUVEC cells. The expression of MMP-2 in co-culture SKOV3/PM4+HUVEC cells and SKOV3/PM4 cells respectively were 1 885 ± 84 and 1 209 ± 114 (P<0.05). But there were no MMP-9 expression in HUVEC cells, SKOV3/PM4 cells, and the co-culture SKOV3/PM4+HUVEC. Conclusion The characteristics of SKOV3/PM4 and HUVEC show significant changes after condition culture and co-culture, it may involve in the microenvironment of the cells and the intercellular crosstalk pathway.

Objective: To investigate the effect of macrophage-capping protein (CapG) on migration and proliferation of human gastric cancer cell line.Methods: Real-time PCR method was used to detect the expression of CapG gene in four gastric cancer cell lines, and AGS cells with low expression and transfection were selected as the research objects.Specific primers were designed for CapG and recombinant plasmids synthesized.A lentivirus packaging system which could express CapG was constructed, and a cell line stably expressing CapG was established by infecting human gastric cancer cell line AGS cells.The effect of overexpression of CapG gene on the growth and proliferation of AGS cells was analyzed by CCK8 assay.Cells cratch and Transwell assay were used to analyze the effect of overexpression of CapG gene on AGS cell migration.Results: After the overexpression of CapG, the growth rate of AGS cells was slightly lower than that of the control group, but there was no significant difference between the two groups (t=2.424, P=0.073).Scratch test showed that the average narrowing distance of the scratches in the CapG experimental group was significantly reduced compared with the control group, the average narrowing distance of the CapG experimental group and the control group was 336.99 μm and 45.54 μm, the difference was statistically significant (t=14.97, P=0.004).The average number of cell penetra-ting membrane in the CapG experimental group and the eGFP control group was 176 and 70, the number of the cells in the CapG experimental group was significantly higher than that of the control group (t=40.00, P<0.001).Conclusion: The overexpression of CapG gene has no significant effect on the growth and proliferation of AGS cells of gastric cancer cell line.Overexpression of CapG gene can promote the migration of AGS cells of gastric cancer cell lines.

OBJECTIVETo explore the effect of Rhodiola on the expression of iNOS mRNA in severe acute pancreatitis (SAP) associated renal injury rats.

METHODSA total of 72 healthy rats were randomly divided into the sham-operated group (S), the SAP associated renal injury group (M), and the Rhodiola-treated group (RHO), 24 in each group. Rats in S and M groups were peritoneally injected with 10 mL/kg saline 3h before modeling, while rats in the RHO group were peritoneally injected with 10 mL/kg Rhodiola Injection 3 h before modeling. The peripheral ligament of pancreas was bluntly dissociated in rats of M and RHO groups. The head of pancreas was occlused by nontraumatic blood vessel forceps 3 h later to establish the model. Eight rats were randomly selected from each group at 12, 24, and 36 h after modeling to detect levels of serum amylase, creatinine, and blood urea nitrogen. Serum levels of interleukin 1β (IL-1β) and interleukin 10 (IL-10) were detected by enzyme-linked immunosorbent assay (ELISA). Pathological changes of the left kidney were observed under light microscope. The expression of inducible nitric oxide synthase (iNOS) mRNA in the right kidney was detected with real time polymerase chain reaction (RT-PCR).

RESULTSCompared with the S group, serum levels of amylase, creatinine (Cr), blood urea nitrogen (BUN), IL-1β, IL-10, and iNOS mRNA expression significantly increased in the M group (P < 0.01). The function of kidney and pancreas were obviously improved in the RHO group than in the M group. Levels of IL-1β and iNOS significantly decreased, but IL-10 levels significantly increased in the RHO group with statistical difference (P < 0.05).

CONCLUSIONRhodiola had better protective effect on SAP associated renal injury, which might be achieved through inhibiting the expression of IL-1β, stimulating the expression of IL-10, down-regulating iNOS mRNA expression, reducing the generation of oxygen free radicals and NO damage to cells, and improving hypoxia tolerance capabilities of the kidney.

Amylases ; Animals ; Blood Urea Nitrogen ; Creatinine ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Interleukin-1beta ; Kidney ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Pancreas ; Pancreatitis ; drug therapy ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Rhodiola

OBJECTIVETo establish a tumor-bearing nude mouse model of human neuroblastoma in order to study the mechanisms of neuroblastoma invasion and metastasis, and to investigate potential therapeutic modalities in the experimental animal models.

METHODSA human neuroblastoma cell line was cultured in vitro. 1 x 10(7) cells undergoing exponential growth were collected in 0.1 ml of suspension and subcutaneously inoculated into the right flank next to the forelimb in nude mice. The biological characteristics of the developed tumors were observed, and histopathological and DNA microarray analyses were performed. The expressions of NSE in the subcutaneous tumor, metastatic tumor and the primary neuroblastoma tumor tissues from a pediatric patient were analyzed by immunohistochemistry.

RESULTSTumors successfully grew in 36 out of 48 injected mice, with a total tumor-formation rate of 75.0%. Metastasis occurred in 10 cases, and the metastatic rate was 20.8%. Tumors in five injected mice grew locally without metastasis. These tumors had large volume and the tumor weight reached up to half of the body weight of the host animal. Four mice exhibited systemic metastasis without tumor growth at the primary inoculation site. There were six mice with locally growing tumor accompanied by metastasis.

CONCLUSIONWe have successfully established a human neuroblastoma xenograft model in nude mice with high tumor growth and metastatic rates. This model depicting the natural cell growth, local infiltration and distant metastasis characteristics of human neuroblastoma, providing an ideal animal model for in vivo studies of neuroblastoma. In addition, the results of this study indicate the heterogeneous nature of neuroblastoma, it may play an important role in metastasis of this tumor.

Animals ; Cell Line, Tumor ; Child, Preschool ; Disease Models, Animal ; Female ; Gene Expression Profiling ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; secondary ; Male ; Mice ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Transplantation ; Neuroblastoma ; genetics ; metabolism ; pathology ; secondary ; Phosphopyruvate Hydratase ; metabolism ; Tumor Burden

Objective To explore the variation and clinical value of the degradation of endothelial glycocalyx in the patients with septic shock. Methods A prospective case control study was conducted. Patients of 18 years or older diagnosed with septic shock and admitted to Department of Critical Care Medicine of Affiliated Hospital of Binzhou Medical University from June 2014 to May 2015 were enrolled. The levels of degradation products, including hyaluronic acid (HA) and heparin sulfate (HS), at 0, 6, 12, 24, 48 hours were determined, while 20 healthy people were enrolled and served as controls. The changes of HA and HS were analyzed in the patients with septic shock. The differences of HA and HS between survival group and death group after 28 days were also analyzed. The relationships between HA, HS and tumor necrosis factor-α (TNF-α), sequential organ failure assessment (SOFA) score, arterial blood lactate (Lac), platelet, albumin were analyzed by Pearson correlation analysis. The receiver-operating characteristic (ROC) curve was plotted to assess the prognostic value of HA and HS for patients with septic shock. Results Thirty-one patients diagnosed as septic shock were enrolled, among whom 17 patients died after 28 days, with a mortality of 54.8%. The levels of HA and HS in patients with septic shock were increased significantly as compared with those of health control group, peaked at 48 hours, and the levels of HA and HS at 48 hours were significantly higher than those at 0 hour [HA (μg/L): 119.47±32.44 vs. 94.84±23.63, HS (μg/L): 72.83±19.03 vs. 58.83±16.63, both P < 0.05]. The levels of HA and HS at 0 hour and 48 hours in death group were significantly higher than those of the survival group [HA (μg/L): 130.42±27.67 vs. 93.29±29.80, 105.14±19.18 vs. 70.82±13.24; HS (μg/L): 67.23±25.01 vs. 39.23±14.58, 79.74±19.84 vs. 56.17±14.53, all P < 0.05]. The levels of HA and HS in patients with septic shock were remarkably positively correlated with the levels of TNF-α, SOFA score, Lac, and platelet, but were remarkably negatively correlated with albumin levels (r value of HA was 0.595, 0.462, 0.545, 0.466, -0.534, respectively; r value of HS was 0.607, 0.468, 0.563, 0.547, -0.455, respectively; all P < 0.05). It was demonstrated by ROC curves that the areas under ROC curve (AUC) of HA and HS at 0 hour and 48 hours for predicting the prognosis of patients with septic shock were 0.881, 0.940 and 0.833, 0.821, respectively, the sensitivities of HA and HS were 87.5%, 100.0% and 83.3%, 81.3%, respectively, and the specificities of HA and HS were 82.6%, 78.3% and 91.3%, 78.3%, respectively. Conclusions The concentrations of degradation products generated by endothelial glycocalyx in the blood of the patients with septic shock are remarkably increased. The elevated levels of the degradation products are closely associated with the severity of septic shock, microcirculation disturbance, and the levels of inflammatory factors.