Objective To observe the effects of tissue factor pathway inhibitor(TFPI) on thrombosis formation in rabbit carotid arteries after ballon injury. Methods Fouty rabbits with the weight 2.5-3.0 kg were respectively divided into 4 groups, Ad-TFPI, Ad-LacZ, PBS and normal control groups. The normal control group was not given any treatment and other 3 groups were given 0.2 ml Ad-TFPI, Ad-LacZ or PBS reproduced by the Dispatch catheter respectively after the PTCA balloon iniury on the right carotid arteries. Ten days after gene transfer the repeated balloon injury was performed in the 3 groups, and the first balloon injury was performed in the normal control group by the same method. The carotid blood flow was recovered immediately after the injury. Thirty minutes later all the animals were sacrificed. The injured carotid arteries and one part of contralateral normal artery were cut down, scissored along the long axis, flattened and fixed in the 2% glutaral. The platelet aggregation and thrombosis formation on the luminal surfaces was observed under electron microscope. Results The electron microscope results showed that the vascular endothelial cell structure was integrated and lined up in order in the nomal artery which had no any injury. After the balloon injury in the normal control group, the structure of the endothelial cell was disintegrated, and there was some platelet aggregation but no fibrosis formation. A large amount of platelet aggregated but no fibrosis formed in Ad-TFPI group after the repeated balloon injury. A large amount of fibrosis formed and red cells piled up in the Ad-LacZ and PBS group. The positive rate of thrombosis formation among groups had siginificant differences(χ2=14.95, P<0.01). The positive rate in Ad-TFPI group(20%) was lower than that in Ad-LacZ group(80%, χ2=7.20, P<0.01) and PBS group(70%, χ2=5.05, P<0.05), but was higher than that in the normal control group(10%, χ2=0.39, P>0.05). The positive rate in Ad-LacZ group(80%) was higher than in the normal control group(10%, χ2=9.90, P<0.01) and in the PBS group(70%, χ2=0.27, P> 0.05). The positive rate in PBS group(70%) was higher than that in the normal control group(10%, χ2=7.50, P< 0.01). Conclusions The repeated balloon injury method can cause a large amount of fibrosis formation in the rabbit carotid. TFPI gene inhibits thrombosis formation in balloon-injured rabbit carotid arteries.

Objective To conduct a computer simulation to evaluate the discrepancy between the cumulative doses calculated by four-dimensional computed tomography (4DCT) images and 4DCT scans (for real-time respiratory motions) due to the patient's irregular breathing.Methods A series of digital phantoms were generated from a patient's 4DCT images to simulate 4DCT images and 4DCT scans (for real-time respiratory motions) resulting from various irregular breathing curves.A six-field intensity-modulated radiotherapy plan was generated.Two cumulative doses in the target were calculated.The first one, named Dall, was calculated by tracking the point displacements in the target manifested on the 4DCT images;the second one, named D4D, was calculated based on the point displacements along the whole breathing motion during 4DCT scanning.Dose discrepancy between D4D and Dall was calculated to evaluate the correlation between breathing pattern and dose discrepancy in the target.Results The dose discrepancy in the target was correlated with mean motion excursion and the standard deviation of motion excursion.ΔDmin(ΔD99) in the target increased from 2.39%(2.04%) to 11.91%(5.24%) as the mean motion excursion increased from 5 mm to 15 mm, and increased from 5.93%(2.15%) to 14.65%(5.01%) as the standard deviation of motion excursion increased from 15% to 45% of the mean motion excursion.When the mean period increased from 3 s to 5 s or the standard deviation of period increased from 10% to 40% of the mean period,ΔDmin(ΔD99) in the target was greater than 6.0%(2.0%), but less than 9.0%(3.0%).When the target diameter was 2 cm, 3 cm, and 4 cm,ΔDminΔD99) in the target was 11.88%(5.50%), 6.91%(2.42%), and 7.53%(3.62%), respectively.Conclusions There is a large discrepancy between the cumulative doses calculated using 4DCT images and 4DCT scans (for real-time respiratory motions) when the patient has irregular breathing.This dose discrepancy depends on mean motion excursion and the standard deviation of motion excursion, but has little relationship with mean period, the standard deviation of period, and tumor volume.

OBJECTIVETo investigate the in vivo gene expression of adenovirus-mediated human tissue factor pathway inhibitor (hTFPI) and its inhibition effects on intimal proliferation in rabbit carotid arteries after balloon injury.

METHODSRabbits underwent carotid artery balloon injuries were treated with Ad-TFPI (n = 25), Ad-LacZ (n = 25) or PBS (n = 10), respectively. Sham operated rabbits (n = 10) serve as normal controls. The expressions of human TFPI at mRNA and protein levels were detected by RT-PCR and ELISA respectively on the 3rd, 7th, 10th, 14th, 28th day after operation. Intimal proliferation was detected by angiograms and morphometric analysis.

RESULTSTFPI mRNA and protein expressions were detected at 3 days and peaked at the 10th and 14th day after TFPI gene transfer. The expressions were still detectable on the 28th day. There was no TFPI expression in Ad-LacZ group. The carotid angiogram results indicated that the minimal lumen diameter in TFPI group was significantly larger and the lumina stenosis percentage was significantly lower in TFPI group compared those in Ad-LacZ and PBS groups (all P < 0.05). The morphometric analysis showed that the intimal area, the ratio of the intimal/media area, the lumina stenosis percentage in TFPI group were all significantly reduced compared with those in Ad-LacZ and PBS groups (all P < 0.01).

CONCLUSIONSThe TFPI gene could be effectively transferred by adenovirus vector to injured carotid arteries and transferred Ad-TFPI could significantly attenuate intimal proliferation in balloon injured carotid arteries in rabbits.

Adenoviridae ; genetics ; Animals ; Carotid Arteries ; metabolism ; Carotid Artery Injuries ; metabolism ; pathology ; Female ; Gene Transfer Techniques ; Genetic Vectors ; Humans ; Lipoproteins ; genetics ; Male ; Rabbits ; Transfection ; Tunica Intima ; pathology

OBJECTIVETo investigate the relationship between the changes of the copy numbers of mtDNA in peripheral blood mono-nucle- ar cell(PBMC) and the disordered of antioxidant capacity of hepatocellular carcinoma (HCC) patients.

METHODSThe Ficoll Hypaque method was used to isolate the PBMC from blood specimens. The ND1 gene of the mitochondrial was amplified by real-time PCR; meantime β-actin was served as a quantitative standard marker; the difference of mtDNA copy number in PBMC was compared between HCC and healthy control group. The level of reactive oxygen species (ROS) in PBMC was determined by flow cytometry. The change of total antioxidant capacity (T- AOC) of plasma was detected by the biochemistry examination.

RESULTSThe copy numbers of ND1 gene in PBMC of HCC was 73% that of the healthy control group,which suggested a decrease of the copy numbers of mtDNA in HCC. The levels of ROS of PBMC in HCC was (417. 82 ± 110.62) and (301.82 ± 75.54) in control group, which showed that the levels of ROS of PBMC in HCC were significant higher than that in control group (P < 0.01).Plasma T-AOC in HCC was (1.30 ± 0.85), and (3.20 ± 1.62) in control. The T-AOC of plasma of HCC was significantly lower than in control group (P < 0.01).

CONCLUSIONThere was a certain relationship between the decrease of the copy numbers of mtDNA and the disordered antioxidant capacity in hepatocellular carcinoma, which may be associated with the development of hepatocellular carcinoma.

Actins ; Antioxidants ; metabolism ; Carcinoma, Hepatocellular ; blood ; genetics ; Case-Control Studies ; DNA Copy Number Variations ; DNA, Mitochondrial ; genetics ; Humans ; Leukocytes, Mononuclear ; metabolism ; Liver Neoplasms ; blood ; genetics ; Reactive Oxygen Species ; metabolism

Methods for determination of heavy metals and harmful residues in traditional Chinese medicine injection were established. Graphite furnace atomic absorption spectrometry was used for determination of lead, cadmium and copper, atomic fluorescence spectrometry for arsenic and mercury. The preprocessing method was optimized. The average recoveries of 5 elements were between 91% and 112% while the precisions were less than 2%. The determination limit of lead, cadmium, copper, arsenic and mercury were 0.28, 0.014, 0.49, 0.19, 0.061 microg x L(-1), respectively. The proposed method was simple, sensitive, accurate and reliable, and could be used widely.
Arsenic ; analysis ; Cadmium ; analysis ; Drugs, Chinese Herbal ; chemistry ; Lead ; analysis ; Mercury ; analysis ; Metals, Heavy ; analysis ; Spectrophotometry, Atomic ; methods

Objective To evaluate the clinical significance of bone scintigraphy in the preoperative diagnosis of sacral tumor. Methods Preoperative 99Tcm-methylene diphosphonate (MDP) whole body bone scintigraphy was performed in total of 103 patients with sacral tumor for whole body survey and radionuclide uptake in the sacral tumor. Of these 103 patients,39 had SPECT. According to the osteoblastic reaction in bone SPECT studies,patterns of tumor with a "hot" lesion was defined as type Ⅰ,a "cold" lesion accompanied with partial uptake was defined as type Ⅱ,a purely "cold" lesion was defined as type Ⅲ,and a "cold" lesion with marginal uptake which produced "doughnut sign" was defined as type Ⅳ. Imaging interpretation was correlated with the final pathologic diagnosis. Results Of the 103 patients,18 ( 17.5% ) had polyostotic involvement. About 46.6% (48/103 ) in planar and 84.6% ( 33/39 ) in SPECT showed decreased uptake at sacrum. Of the bone metastatic patients (n =21 ) ,12 (51.7%) had sole metastasis to sacrum. Tumor with type Ⅰ (6/6) or type Ⅱ (16/19) uptake was likely to be a malignancy,whereas type Ⅲ uptake tended to occur in the benign disease in those patients without polyostotic involvement( 5/7 ),and type Ⅳ was all appeared in giant cell tumors( n = 5 ). Conclusions Preoperative bone scintigraphy is useful in examination of polyostotic involvement for the patients with sacral tumor,but it is limited for diagnosing isolated sacral metastatic disease. Tumor uptake on bone scintigraphy can be helpful in differential diagnosis of sacral tumor.

High mobility group box 1 (HMGB1) protein is released by apoptotic cells and monocytes／macrophages.It plays an important role in mediating inflammation development and promoting tissue repairment and regeneration via combining with corresponding receptors .In recent years ,some researches indicated that HMGB1 possessed im-portant diagnostic and prognostic assessment value for coronary heart disease ,heart failure and myocardial injury etc .The present article summarized research progress of HMGB1 in cardiovascular diseases in recent years .

OBJECTIVETo evaluate the relationship between circulating levels of insulin-like growth factor-1 (IGF-1), IGF-binding protein-3 (IGFBP-3) and colorectal cancer.

METHODSA meta-analysis of 6 epidemiological studies on insulin-like growth factors and risk of colorectal cancer were performed.

RESULTSThe pooled odds ratio (OR) of IGF-1 and IGFBP-3 were 1.56 (95% CI: 1.14-2.13) and 0.78 (95% CI: 0.43-1.44) respectively. According to the results from different measurements (enzyme-linked immunoabsorbent assay and immunoradiometric assay), the pooled OR were 1.92 and 1.23 for IGF-1, 0.46 and 1.44 for IGFBP-3 respectively.

CONCLUSIONHigh serum levels of IGF-1 were independent risk factors of colorectal cancer but the OR of IGFBP-3 was not statistically significant. The heterogeneity between studies on IGFBP-3 and colorectal cancer was caused by different measurements used, but there was still a need to conduct simultaneous large size study under 2 different measurements for further conclusion.

China ; epidemiology ; Colorectal Neoplasms ; blood ; epidemiology ; Enzyme-Linked Immunosorbent Assay ; methods ; Insulin-Like Growth Factor Binding Protein 3 ; blood ; Insulin-Like Growth Factor I ; metabolism ; Radioimmunoassay ; Risk Factors

BACKGROUNDIndoleamine-2,3-dioxygenase (IDO) is proven to suppress hepatitis B virus (HBV) specific immune response and depletion of IDO may be a useful approach for HBV therapy. To test this concept, we constructed recombinant adenovirus with human IDO and HBV preS, which would form the basis for future in vivo experiments.

METHODSThe fragment of human IDO and HBV preS cDNA were subcloned into multiple cloning sites in an adenoviral vector system containing two cytomegalovirus (CMV) promoters. Recombination was conducted in the Escherichia coli BJ5183. The recombinant adenovirus containing hIDO gene and HBVpreS gene was packaged and amplified in 293 cells. Integration was confirmed by polymerase chain reaction as well as the quantification of viral titers. HepG2 cells were infected with the recombinant adenovirus and mRNA and protein specific for hIDO and HBVpreS was detected by RT-PCR and Western blotting respectively.

RESULTSThe recombinant adenovirus was produced successfully. Its titer was 2.5 × 10(9) efu/ml. IDO and HBVpreS mRNA as well as the encoded proteins could be found in transfected HepG2 cells, but not in control HepG2 cells.

CONCLUSIONThe transfer of hIDO-HBVpreS with double-promoter adenoviral vector was efficient. The recombinant adenovirus with hIDO and HBV preS would provide the experimental basis for future studies.

Adenoviridae ; genetics ; Cloning, Organism ; Genetic Vectors ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; genetics ; Recombination, Genetic

OBJECTIVETo test the effect of asarinin, the extract of Herba Asari, on the acute heart transplantation rejection and the expression of adhesion molecule.

METHODAsarinin was extracted from herba asari. 64 SD rats undergone heart transplantation were divided into four groups: group A (control group), group B (Cyclosporine A treated), group C (Asarinin treated), and group D (1/2 CsA and 1/2 Asarinin). Some rats were used to examine survival time (n = 8) and the others were used to observe the pathological injury and the expression level of interrellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-I (VCAM-1) by using immunohistochemistry.

RESULTAsarinin could prolong the survival time of allografts, which was similar to CsA group (P > 0.05). Asarinin could relieve the damage of cardiomyocytes of the transplanted. Asarinin could also decrease the level of ICAM-1 and VCAM-1 in the allografts.

CONCLUSIONAsarinin may play important roles in suppressing the immune rejection, prolong the allografts survival time and protect the donor organ, which was similar to CsA. The expression level of ICAM-1 and VCAM-1 is increased in suppressing the course of acute rejection and asarinin can inhibit their expression level. Asarinin can decrease the dosage of CsA.

Animals ; Asarum ; chemistry ; Dioxoles ; isolation & purification ; pharmacology ; Graft Rejection ; metabolism ; Graft Survival ; drug effects ; Heart Transplantation ; Intercellular Adhesion Molecule-1 ; metabolism ; Lignans ; isolation & purification ; pharmacology ; Male ; Myocardium ; metabolism ; pathology ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Vascular Cell Adhesion Molecule-1 ; metabolism