Angiopoietins ; metabolism ; physiology ; Animals ; Cell Proliferation ; Endothelial Cells ; cytology ; physiology ; Humans ; Neoplasms ; blood supply ; Neovascularization, Pathologic ; physiopathology ; Neovascularization, Physiologic ; physiology ; Pericytes ; cytology ; metabolism ; physiology ; Proto-Oncogene Proteins c-sis ; metabolism ; physiology ; Receptor, Platelet-Derived Growth Factor beta ; metabolism ; physiology ; Receptor, TIE-2 ; metabolism ; physiology ; Signal Transduction

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Emodin ; pharmacology ; Humans ; Neoplasms ; pathology ; Tumor Cells, Cultured

Objective To study the influence of low-iodine diet on the expression of homeobox gene nkx2.1 in rat cerebral tissue. Methods Forty female Wistar rats were randomly divided into two groups according to body. quality: low-iodine group and control group,both fed with low-iodine feed at an iodine content of 13.66 μg/kg,respectively given the deionized water and 200 μg/L KIO3 solution. The hormone levels of two group rats were determined with chemiluminescence immunoassay after three months, and then mated with healthy male rats. Cerebral tissues were taken from the fetus of 16-day pregnancy,newborn and 20 days old offspring in low-iodine and control group to detect the content of nkx2.1 mRNA using real-time fluorescence quantitative PCR (FQ-PCR) techniques. Results Serum TT3, TT4, FT3, FT4 level of rats in low-iodine group(0.89±0.20, 0.32±0.16, 3.33± 0.61, 3.28±0.80) was respectively lower than that in the control group(1.04±0.06, 39.42±14.68,4.83±0.33, 26.99±4.48;t = 2.71,6.52,5.70, 12.89, P < 0.05 or < 0.01). The relative nkx2.1 mRNA expression was(5.60± 0.30)×10-3, (1.20 ± 0.29)×10-3, (0.18± 0.06)×10-3 respectively in the fetus of 16-day pregnancy, newborn and 20 days old offspring of control group, while it was (3.00 ± 0.55)×10-3, (1.90 ± 0.21)×10-3,(0.69 ± 0.15)×10-3 in the low-iodine group. The difference of nkx2.1 mRNA expression was significant among fetal and neonatal rats in the control group and low-iodine group(F = 210.07,162.40, both P < 0.01). The nkx2.1 mRNA expression of newborn rats was lower than that of 16-day pregnancy in both groups(P < 0.01), and that of 20 days old rats was lower than that of 16-day pregnant and neonatal rats(P < 0.01). The 16-day pregnant rats of control group had obviously higher level of nkx2.1 expression than those in the low-iodine group(t = 16.073, P< 0.01), while the nkx2.1 of newborn and 20 days old low-iodine rats expressed much higher than healthy rats(t = 7.573,12.221, P < 0.01). Conclusions Brain development retardation caused by low-iodine is closely related to nkx2.1 differential expression in the brain tissue.

Objective To study the influence of low-iodine diet on the expression of homeobox gene Nkx-6.1 and Nkx-6.2 in rat cerebrum tissue, and to explore the possible molecular mechanism of cerebrum development retardation caused by low-iodine. Methods Twenty female Wistar rats were randomly equally divided into two groups: low-iodine group and control group, both fed with low-iodine diet as low as 13.66 μg/kg determinated by spectrophotometry in Tianjin Institute of Endocrinology and the former with deionized water, the later 200 μg/L potassium iodate. Thyroid hormone level was detected using chemiluminescence immunoassay 3 months later and they were mated with male rats normally fed. Rats of 16-day pregnancy, new-born and 20th days old were detected the content of Nkx-6.1 and Nkx-6.2 mRNA in the cerebrum tissue by real-time fluorescence quantitative PCR 0.61), (3.28±0.80)pmol/L] were lower than the control group[(1.04±0.06), (39.42±14.68)nmol/L, (4.83±0.33), day pregnancy, new-born and 20th days old of control group was (1.90±0.23)×10-3,(1.86±0.40)×10-4, (1.11± 0.27)×10-4(F=827.58, P<0.01), Nkx-6.1 mRNA expression level gradually decreased along with aging(all P<0.05). The intra-group difference was significant (F=297.25, P<0.01) and the Nkxr.1 mRNA expression level during 16 days of pregnancy was the highest(P<0.01). It was higher in the control group than in the low-iodine group during 16 days of pregnancy (t=10.14, P<0.01) as well as in the low-iodine group than in the in 16-day pregnancy, new-born and 20th days old of control group was respectively(1.03±0.19)×10-2, (1.33± 0.10)×10-3, (8.79±0,87)×10-3, and that of low-iodine group was (0.31±0.03)×10-2, (1.53±0.13)×10-3, (7.51±0.86)×10-2. The intra-group difference was significant(F=1293.02,1065.83, all P<0.01). Nkx-6.2 expression level during 20th days old was the highest(P<0.01) and that of newborn was the lowest(P<0.01). The Nkx6.2 mRNA expression level in control group were higher than the low-iodine group during 16-day pregnancy and 20th days old(t=14.35, 4.05, all P<0.01). It was higher in the low-iodine group than in the control group during newboru(t=4.78, P<0.01). Conclusions The difference in the expression of Nkx-6.1 and Nkx-62 is highly related to the brain development retardation caused by low-iodine.

Objective To detect the levels of five trace elements in whole blood of patients with Keshan disease(KSD) and dilated cardiomyopathy(DCM) and explore their role in the pathogenesis of KSD.Methods One hundred and four patients with chronic KSD were selected from Keshan diseased areas in Shandong,Sichuan and Inner Mongolia.Thirty patients with DCM were selected from Qilu Hospital of Shandong University,Jinan Central Hospital,The First People's Hospital.Ninety-one healthy people from KSD endemic areas and 39 healthy people from Jinan were selected as endemic healthy controls and non-endemic healthy controls,respectively.Blood samples were collected to determinate the level of selenium (Se),copper (Cu),zinc (Zn),chromium (Cr) and manganese (Mn) with fluorescence method and atomic absorption spectrometry,according to the principle of informed consent.Results The level of Se,Zn and Cr of KSD group[(36.0 + 4.9)μg/L,(22.73 + 4.62)mg/L,(0.56 + 0.17)mg/L] was significantly lower than that of non-endemic healthy controls [(56.4 ± 6.8)lμg/L,(25.35 ± 4.44)mg/L,(0.71 ± 0.17)mg/L,all P ＜ 0.05],but the level of Cu of KSD group[(0.95 ± 0.24)mg/L] was significantly higher than that of non-endemic healthy controls[(0.73 ± 0.13) mg/L,all P ＜ 0.05].The level of Se and Cr of KSD was significantly lower than that of endemic healthy controls[(54.5 ± 5.4)μg/L,(0.87 ± 0.02)mg/L,P ＜ 0.05],and Cu was significantly higher than that of endemic healthy controls[(0.66 ± 0.02)mg/L,P ＜ 0.05].The level of Cu and Zn of KSD was significantly lower than that of DCM [(1.21 ± 0.23)mg/L,(27.09 ± 7.10)mg/L,all P ＜ 0.01].The level of Se and Cr of DCM group[(39.6 ± 3.5)μg/L,(0.58 ± 0.14)mg/L] was significantly lower than that of non-endemic healthy controls(all P ＜ 0.01),but Cu[(1.21 + 0.23)mg/L] was significantly increased (P ＜ 0.01).Compared with non-endemic healthy controls,the level of Se of endemic healthy control group was significantly decreased (P ＜ 0.01),while Cu was significantly increased (P ＜ 0.01).Se,Zn and Cr level of KSD decreased gradually following elevated heart function level,but the level of Cu gradually increased.Conclusions The metabolism of Se,Cr,Cu and Zn is unbalanced in KSD patients,whose Se level is still lower than that of people in non-endemic areas.The change of Se,Cr,Cu and Mn level between KSD and DCM is consistent.

The aim of this study was to detect the rate of T-helper (Th)17 cells and interleukin (IL)-17 level in peripheral blood of patients with primary immune thrombocytopenia (ITP) and to explore their clinical significance. The proportion of Th17 cells from 48 patients with ITP and 28 healthy controls was detected by flow cytometry, and the IL-17 level was evaluated by enzyme-linked immunosorbent assay (ELISA). The results showed that the percentage of Th17 cells in ITP group was (1.40 ± 1.35)%, which was significantly higher than that in healthy control group (P < 0.05), but in the glucocorticoid hormone-treated group it was significantly lower than that in treated group without glucocorticoid hormone(P < 0.05). The level of IL-17 expressed by Th17 cells in ITP patients was (19.624 ± 5.187) pg/ml, which was higher than that in the healthy control group (P < 0.05), it was lower in the glucocorticoid hormone treated group than that in treated group without glucocorticoid hormone, but there was no statistically significant difference between the glucocorticoid treated and treated group without glucocorticoid hormone (P > 0.05). It is concluded that the Th17 cells may involve in the pathogenesis of ITP, and the glucocorticoid hormone probably plays a therapeutic role through inhibiting Th17 cells.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Interleukin-17 ; metabolism ; Male ; Middle Aged ; Th17 Cells ; metabolism ; Thrombocytopenia ; drug therapy ; metabolism ; Young Adult

This study was purposed to investigate the correlation of deep vein thrombosis (DVT) with C-reactive protein (CRP), fibrinogen (Fg), coagulation factor VIII (FVIII:C), coagulation factor IX (FIX:C) and to explore the effect of inflammation and coagulation as well as their interaction in DVT and its mechanism. 59 patients with DVT undergoing selective venous ultrasonography and 26 healthy individuals as controls were enrolled in this study. The plasma level of CRP was detected by immunoturbidimetry, FVIII:C, FIX:C levels were determined by a one-stage assay and fibrinogen level was measured by full-automatic biochemical apparatus. The results showed that the mean levels of plasma CRP, Fg, FVIII:C and FIX:C were significantly higher in deep vein thrombosis group than that in controls [CRP (2.67 +/- 0.91) vs (0.14 +/- 0.08) mg/dl; Fg (4.73 +/- 1.36) vs (2.79 +/- 0.66)g/L; FVIII:C (126.71 +/- 28.10) vs (81.35 +/- 20.77)%; FIX:C (81.01 +/- 23.60) vs (70.71 +/- 11.3)%] (p < 0.01), and the level of plasma CRP was strongly correlated with Fg, FVIII:C and FIX:C (r(s) = 0.432, 0.571 and 0.544, p < 0.01). It is concluded that the DVT and inflammation are closely related, increased level of plasma CRP may be a predictor of DVT. Increased plasma levels of Fg, FVIII:C and FIX:C all are important risk factors to DVT. Interaction between inflammation and coagulation promote the incidence of DVT, which may be one of DVT pathogenesis.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biomarkers ; blood ; Blood Coagulation ; C-Reactive Protein ; metabolism ; Case-Control Studies ; Factor IX ; metabolism ; Factor VIII ; metabolism ; Female ; Fibrinogen ; metabolism ; Humans ; Inflammation ; Male ; Middle Aged ; Venous Thrombosis ; blood ; etiology ; Young Adult

OBJECTIVETo explore the effect of N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide(W7) on the differentiation from human adipose-derived mesenchymal stem cells (hADSCs) to endothelial cells.

METHODShADSCs were cultured with serum-free differential medium containing 40 ng/ml vascular endothelial growth factor (VEGF) and 10ng/ml basic fibroblast growth factor (bFGF). Cells were divided into control group (differential medium without W7), high-dose group (containing 30 μmol/L W7), medium-dose group (containing 20 μmol/L W7), and low-dose group ( containing 10 μmol/L W7). The hADSCs were cultured for 8 days, and then the changes in the phenotypes of von Willebrand factor (vWF) and vessel-selective cadherin (VE-Cadherin) were detected by flow cytometry (FCM). The intracellular Ca(2+) labeled with Fluo-3 was detected by laser confocal microscopy. After hADSCs planting on Matrigel, their angiogenic potentials were observed under the inverted phase contrast microscope, and the expression of extracellular regulated kinase (ERK) and phosphorylated extracellular regulated kinase (p-ERK) were evaluated by Western blot.

RESULTSAfter the hADSCs were cultured for 8 days, compared with the control group, the expressions of vWF and VE-Cadherin significantly increased along with the decrease of W7 level and the intracellular Ca(2+) also significantly increased (Pü0.01). Lumina-like vascular structure was formed in W7 treatment groups, but not in the blank control group. Compared with the blank control group, the expression of ERK showed no significant in W7 treatment groups (high-, medium-, and low-dose groups)(P>0.05); however, along with the decrease of W7 levels, the expression of p-ERK significantly increased(P<0.05).

CONCLUSIONW7 in proper levels can effectively induce the differentiation from hADSCs to endothelium by increasing the intracellular Ca(2+) level and thus activating the ERK/MAPK pathway.

Adipose Tissue ; cytology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Sulfonamides ; pharmacology

OBJECTIVETo study the effect of co-blockage of vascular endothelial growth factor (VEGF) and its receptor (KDR) on growth of bladder carcinoma T24 cells and nude mice xenograft.

METHODST24 cell line co-transfected with VEGF siRNA and sKDR expression plasmids was developed and its proliferation was assayed by MTT and apoptosis by FCM. The nude mice model bearing bladder carcinoma xenograft was established. The tumor cell VEGF expression, stroma microvessel density (MVD) and tumor cell topoisomerase II alpha (Topo II alpha) expression were detected by immunohistochemistry. Cell apoptosis was estimated by TUNEL assay.

RESULTSMTT assay showed that cell proliferation in VEGF siRNA, sKDR and combination groups was 56.3% +/- 8.3%, 42.6% +/- 13.8% and 32.5% +/- 4.3%, respectively, significantly lower than that in the scramble control (97.3% +/- 11.6%, P < 0.0001). FCM showed there were sub-diploid apoptotic peaks before G1 phase in VEGF siRNA, sKDR and combination groups, and apoptosis ratio was 5.1% +/- 0.9%, 4.2% +/- 0.5% and 8.8% +/- 0.7%, respectively, all of which were higher than that in the scramble control (0.9% +/- 0.4%, P < 0.05), and the combination group had even more higher apoptosis than the two singlely treated groups (P < 0.01). In vivo test showed that tumor growth was inhibited in VEGF siRNA, sKDR and combination groups, and from day 16 the tumor volume in combination group was significantly smaller than that in scramble control (P < 0.05), and from day 28 the tumor almost lost the ability to further growth. Immunohistochemistry revealed VEGF expression in combination group was 54.37 +/- 5.28, significantly lower than that in the scramble control (141.66 +/- 8.59, P < 0.0001). MVD number was only 8.22 +/- 3.79, much less compared with that in the scramble control (61.76 +/- 5.28, P < 0.0001) or sKDR group (19.46 +/- 4.16, P = 0.0089). Tumor cell proliferation index in the combination group (1.5% +/- 0.7%) was significantly decreased compared with that in the scramble control (11.8% +/- 5.2%, P < 0.0001), and apoptosis index (67.2% +/- 8.5%) was much higher than that in the scramble control (8.7% +/- 2.7%, P < 0.0001), VEGF siRNA group (54.3% +/- 4.8%, P = 0.0492) or sKDR group (52.3% +/- 6.4%, P = 0.0293).

CONCLUSIONVEGF siRNA or sKDR alone can inhibit tumor cell proliferation and induce cell apoptosis, but co-blockage of VEGF and KDR by their combination shows more significant therapeutic efficacy.

Animals ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; prevention & control ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Tumor Burden ; Urinary Bladder Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism ; Xenograft Model Antitumor Assays

OBJECTIVETo study the number of CECs in patients with acute myocardial infarction (AMI) and unstable angina (UA), and to investigate its relationship with inflammatory related cytokines.

METHODS37 patients with AMI, 12 patients with UA, and 42 health controls were studied. CECs were isolated from peripheral blood by using of immunomagnetic beads coated with antibodies against CD146. Their endothelial origin was confirmed by the positive labelling of von Willebrand Factor (vWF), CD31 and electron microscope. Annexin V-FITC/PI kit was used to measure the apoptosis of CECs. Inflammatory related cytokines were analyzed turbidimetrically or ELISA using of commercially available testing kit.

RESULTSCECs number was significantly higher in AMI and UA patients [medians (interquartile range) were 52 (28 approximately 81.5) cells/ml and 29 (18 approximately 61) cells/ml respectively] compared with health control [10.5 (6-16.5)cells/ml, P < 0.001]. After excluding diabetes patients, the number of CECs and CRP in AMI and UA group (n = 26) were still significantly higher than controls. The necrotic rate of CECs in AMI and UA was significantly higher than controls (P < 0.01). Correlation analysis revealed a significant positive correlation between CECs and CRP, or IL-6 (r = 0.677, 0.316, P = 0.000, 0.002). The multivariate linear regression analysis showed that CRP and Diabetes increased the number of CECs significantly (OR = 0.620, 0.164, 95% CI 3.985-6.751, 0.301-21.877, P = 0.000, 0.044).

CONCLUSIONThe mechanism responsible for the increase of CECs in acute coronary disease may be due to the vessel injury caused by inflammation.

Acute Coronary Syndrome ; blood ; Aged ; Angina, Unstable ; blood ; C-Reactive Protein ; metabolism ; Case-Control Studies ; Endothelial Cells ; cytology ; Endothelium, Vascular ; chemistry ; cytology ; Female ; Humans ; Inflammation ; Interleukin-6 ; metabolism ; Linear Models ; Male ; Middle Aged ; Multivariate Analysis ; Myocardial Infarction ; blood ; Tumor Necrosis Factor-alpha ; metabolism