BACKGROUND: Previous studies demonstrated that smooth muscle injury or ischemia/reperfusion injury result in tissue factor (TF) increasing. However, few reports concern the expression and mechanism of TF in venous bypass grafting.OBJECTIVE: To examine changes in TF protein expression in response to venous bypass grafting.DESIGN, TIME AND SETTING: The animal observation experiment was performed at the Department of Cardiovascular Surgery, Affiliated Union Hospital of Tongji Medical College, Huazhong University of Science and Technology from May 2006 to May 2008.MATERIALS: A total of 30 Sprague-Dawley (SD) rats.METHODS: Rats were underwent interposition bypass grafting of the common carotid artery via the ipsilateral external jugular vein. Namely, the proximal end of external jugular vein was ligated at the joints of external jugular vein and internal jugular vein, and the distal end of external jugular vein was ligated before branches. The proximal and distal ends of common carotid artery were occluded by artery clamp, and a 5 mm artery was removed. The proximal end of artery was anastomosed with distal end of artery, and the frontal wall was sutured with posterior wall. After that, the proximal end of external jugular vein was cut down and coincided with the distal end of common carotid artery.MAIN OUTCOME MEASURES: The expression of TF and proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry. Meantime, TF activity in vessel protein extracts was determined with TF activity assay kit, and the thickness of intima, media were calculated by computer imaging analysis system. The contralateral external jugular veins were served as the control.RESULTS: The adventitia of all vessels showed abundant TF staining. In early vein grafts, TF staining was markedly increased in the intima and media. However, intimal and media TF staining was absent in the contralateral control jugular veins and late vein grafts. The number of PCNA positive cells was increased in the vein grafts at day 3 after grafting, obvious increased at day 7, and reached the peak at day 14. TF activity in whole-vessel protein extracts was similar in control veins and early and late vein grafts. The thickness of neointima of the vein graft increased significantly at days 7, 14, and 28, and the thickness of media increased significantly at days 14 and 28.CONCLUSION: The changes of TF expression at various time points may relate to hyperplasia of neointima.

Induced pluripotent stem cells (iPSCs) can be propagated indefinitely, while maintaining the capacity to differentiate into all cell types in the body except for the extra-embryonic tissues. This iPSC technology not only represents a new way to use individual-specific stem cells for regenerative medicine but also constitutes a novel method to obtain large numbers of disease-specific cells for biomedical research. However, the low efficiency of reprogramming and genomic integration of oncogenes and viral vectors limit the potential application of iPSCs. Chemical-induced reprogramming offers a novel approach to generating iPSCs. In this study, a new combination of small-molecule compounds (SMs) (sodium butyrate, A-83-01, CHIR99021, Y-27632) under conditions of transient folate deprivation was used to generate iPSC. It was found that transient folate deprivation combined with SMs was sufficient to permit reprogramming from mouse embryonic fibroblasts (MEFs) in the presence of transcription factors, Oct4 and Klf4, within 25 days, replacing Sox2 and c-Myc, and accelerated the generation of mouse iPSCs. The resulting cell lines resembled mouse embryonic stem (ES) cells with respect to proliferation rate, morphology, pluripotency-associated markers and gene expressions. Deprivation of folic acid, combined with treating MEFs with SMs, can improve the inducing efficiency of iPSCs and reduce their carcinogenicity and the use of exogenous reprogramming factors.

This study aimed at evaluating the quality of the precise powder decoction pieces (PPDP) of L.japonicae Flos (LJF) compared with the traditional commercial slices with chemical fingerprint methods and DNA molecular identification technology.Different specifications of PPDP were prepared,their dry extract contents were in contrast with that of commercial slices.The three batches of commercial slices were collected,and the content uniformity,fingerprint and similarity evaluation before and after the mixing and pulverization were studied by HPLC-DAD and DNA sequence alignment.As a result,the paste rate of PPDP was slightly higher than that of the traditional commercial slices.The dissolution of chlorogenic acid of PPDP was higher than that of the traditional commercial slices.RSD of inter-assay dissolutions of chlorogenic acid of commercial slices was 11.93％,which was reduced to 8.29％ after mixing and preparing into PPDP.The fingerprint showed that the slimilarity of the fringerprint of the mixed and powdered LJF was elevated with 7 common peaks.All the common peaks were increased at different levels.In conclusion,compared with traditional commercial slices of LJF,PPDP apparently improved the dissolution rate and the quality uniformity,indicating that the boiled powder of CRP obviously presented vantages in clinic.

A anaerobic hydrogen-producing strain HR-1 was isolated from compost. Phylogenetic analysis based on 16S rRNA sequence similarity indicates that strain HR-1 is the closest relative to Clostridium ace- tobutylicum ATCC 824, with the similarity of 96%. Biological characteristics and phylogenetic analysis of 16S rRNA gene indicate that HR-1 is a new species named Clostridium sp. HR-1. Cells are Gram-positive, mobile rod-shaped. Spores and flagellums were no observed. Temperature range for growth is 10?C to 45?C (optimum temperature 37?C~39?C), and range pH for growth is 4.0 to 10.0 (optimum pH 7.5~8.0). H2, CO2, acetate, butyrate and a little ethanol are the end products of PYG fermentation. Strain HR-1 has the ability to use organic nitrogen and inorganic nitrogen sources for growth and hydrogen production, and yeast extract is the optimum nitrogen source for hydrogen production. Strain HR-1 produces hydrogen from xylose (3 g/L) at 37?C and initial pH 6.5, the hydrogen yields and maximal hydrogen production rate are 1.84 mol-H2/mol-xylose and 10.52 mmol-H2/h?g-cdw, respectively. Strain HR-1 is able to utilize glucose, galactose, fructose, mannose and cellobiose for hydrogen production and the hydrogen yields from glucose is 2.36 mol-H2/mol-glucose.

Objective To compare the results of mitral valve reconstruction and replacement as treatments for moderate to severe ischemic mitral regurgitation(IMR), and report the mid-term outcome. Methods From June 2002 to May 2008, 83 pa-tients with moderate IMR(35 cases) and severe IMR (48 cases) underwent coronary artery bypass grafting(CABG) combined with mitral valvuloplasty (MVP) (n = 43) or mitral valve replacement (MVR) (n = 40). There were 49 males and 34 females with a mean age of (59.3±7.5) years(51 -77years). The procedures of MVP included annuloplasty with a Dacron or autologous per-icardium ring in 21cases, commissural annuloplasty in 9, quadrangular resection of the posterior leaflet in 9 and using St. Jude mitral annuloplasty ring in 4. In the cases underwent MVR, 28 patients received mechanical prostheses and 12 received biopros-theses. Results 30-day mortality rate was 2.3% for MVP and 5.0% for MVR (P >0.05). The 30-day complication rate was similar for the 2 groups but mechanical ventilation time was longer for MVR patients. Mild MR ocurred in 6 patients with MVP (P <0.05). Sevonty-six patients were followed by outpatient department visit or telephone for (20.2 ± 4.9) months (3 - 60 months). During the follow-up period, 7 patients with MVP had mild insufficiency but free off etber complications. All the valve prothesis functioned well. However, 3 cases had thromboembolic complications and 7 late deaths were recorded in MVR group. Five-year complication-free survival rate was 90% for MVP group and 61% for MVR. Conclusion MVP resulted in excellent durability and provided significant mid-term survival benefit over MVR. MVP should be the first choice for patients with chronic IMR.

Objective To investigate the significance of the serum levels of interleukin-10 (IL-10),IL-13,IL-15 of patients with chronic hepatic failure and the correlation between those inter- leukin levels and nosocomial infections.Methods The serum levels of IL-10,IL-13,IL-15 of 58 patients with chronic hepatic failure were measured by double antibody sandwich enzyme-linked immu- nosorbent assay at the time of admission and 2 weeks after admission.Results The serum levels of IL-15 and the propotion of IL-15/IL-10 and IL-15/IL-13 in patients with chronic hepatic failure group at the time of admission were significantly higher than those in healthy control group[(358.16?290.91) ng/L vs (38.55?21.49) ng/L,12.93?14.26 vs 1.10?0.55,98.55?97.5.5 vs 9.70?5.03,respectively,all P=0.000].Those in death group were significantly higher than those in improving group[(479.93v205.52) ng/L vs (244.51?236.29) ng/L,17.65?17.78 vs 8.53?7.98,130.69?115.50 vs 68.55?65.99,respectively,all P

BackgroundDiabetic retinopathy (DR) is the result of the cytokine network disorders,the imbalance of angiogenic factor and vascular inhibitory factor is the start factor.ObjectiveTo analyze the levels of CXC chemotatic factors of type 2 diabetes mellitus patients,evaluate the clinical application value of them in different clinical types of DR using receiver operating characteristic (ROC)analysis and to approach the new way of individualized treatment.Methods This was a prospective research.The gold standard was ophthalmolscope and fundus fluorescein angiography.The levels of CXC chemotatic factors and multiplicaiton factors were measured in 96 cases with type 2 diabetes mellitus (66 cases with retinopathy and 30 cases without retinopathy as control).The assessment tasks were performed for these index and courses of DR with ROC curve.Results The expression of age,course of disease has significant difference in different courses of DR ( F =8.507,P =0.001 ; F =28.143,P =0.000).Compared with the control group,the expression of growth-related oncogene-α ( GROα ) ( t =- 2.172,P =0.035,AUC =0.625 ),whole blood viscosity 200 ( t =- 3.724,P =0.001,AUC =0.904 ) and neutrophilic leukocyte (t=-2.562,P =0.013,AUC =0.577 ) has significant difference in the group of mild NPDR.Compared with the control group,the expression of interferon-γ-inducible protein 10 ( IP-10 ) ( t =-3.591,P =0.001,AUC =0.592 ),platelet derivation growth factor-BB ( PDGF-BB ) ( t =- 3.233,P =0.003,AUC =0.735 ),vascular endothelial growth factor(VEGF) ( t =- 3.617,P =0.001,AUC =0.776 ),C peptide ( t =- 3.366,P =0.002,AUC =0.962 ),leukocyte ( t=-3.201,P =0.003,AUC =0.852) and neutrophilic leukocyte(t =-4.201,P=0.000,AUC =0.852) has significant difference in the group of moderate and severe NPDR.ConclusionsCXC chemotatic factors may act as reactivator in the pathogenesis of DR,GROα and IP-10 may be useful for clinical monitoring of the severity of DR,and evaluating the imbalance state of chemotatic factors maybe a new approach to clinical monitoring and prognosis of DR.

Objective To study the tropism of umbilical cord mesenchymal stem cells (UCMSCs) to glioma stem cells and their mechanism.Methods The isolated humanized UCMSCs were cultured and identified.The glioma stem cells were induced to remain at the resting, proliferation or differentiation states by soft or hard agar combined with stemness maintenance factors (epidermal growth factor [EGF] and basic fibroblast growth factor [bFGF]): soft agar+EGF+bFGF group, hard agar+EGF+bFGF group, and hard agar group;clone spheres of the stem cells were cultured for 7 d, and then, the growth curve of clone spheres diameters was drawn;flow cytometry was used to detect the cell cycle of the stem cells 72 h after culture;CD133, Ki67 and GFAP expressions in the clone spheres of stem cells in the three groups were observed by immunofluorescent staining;the tropism aptitude of UCMSCs to glioma stem cells at different states was detected by Transwell dual chamber culture system;the protein expression levels of stem cell factor (SCF), stromal-derived factor (SDF)-1 and vascular endothelial growth factor (VEGF) in glioma stem cells were monitored by enzyme immunoassay (ELISA);Western blotting was used to measure the protein expression levels of c-Kit, C-X-C chemokine receptor type 4 (CXCR-4) and VEGF receptor in UCMSCs.Results The diameters of clone spheres in the hard agar+EGF+bFGF group and hard agar group were significantly larger than those in the soft agar+EGF+bFGF group on the second and 7th d of culture (P＜0.05);cell cycle analysis indicated that glioma stem cells in the soft agar+EGF+bFGF group, hard agar+EGF+bFGF group, and hard agar group were successfully remained at the resting, proliferation or differentiation states;high CD133 expression ([95.79±5.31] ％) was noted in the cells of soft agar+EGF+bFGF group, high Ki67 and GFAP expressions ([89.39±7.45 and 63.49±16.54] ％) were noted in the cells of hard agar+EGF+bFGF group, and high GFAP expression ([97.36±3.48] ％) was noted in the cells of hard agar group;Transwell assay indicated that the migration percentage of cells from the hard agar+EGF+bFGF group was significantly higher than that in the soft agar+EGF+bFGF group, hard agar group and two control groups (P＜0.05).higher protein expression levels of SCF, SDF-1 and VEGF in the hard agar+EGF+bFGF group were noted as compared with those in the soft agar+EGF+bFGF group and hard agar group (P＜0.05).UCMSCs were induced to express significantly higher levels of c-Kit, CXCR4 and VEGF receptor in the hard agar+EGF+bFGF group as compared with those in the soft agar+EGF+bFGF group and hard agar group (P＜0.05).Conclusion SCF/c-Kit, SDF-1/CXCR4 and VEGF/VEGFR axis expression defects might be the potential mechanism of the migration defect of UCMSCs to glioma stem cells at the resting state.

OBJECTIVE: The clinical and pathological characteristics of 10 cases of cerebral amyloid angiopathy (CAA)-related cerebral lobar hemorrhage (CLH) that was diagnosed at autopsy were investigated to facilitate the diagnosis of this condition. METHODS: The clinical characteristics of 10 cases of CAA-related CLH were retrospectively reviewed, and a neuropathological examination was performed on autopsy samples. RESULTS: The 10 cases included two with a single lobar hemorrhage and eight with multifocal lobar hemorrhages. In all of the cases, the hemorrhage bled into the subarachnoid space. Pathological examinations of the 10 cases revealed microaneurysms in two, double barrel-like changes in four, multifocal arteriolar clusters in five, obliterative onion skin-like intimal changes in four, fibrinoid necrosis of the vessels in seven, neurofibrillary tangles in eight, and senile plaques in five cases. CONCLUSION: CAA-related CLHs were located primarily in the parietal, temporal, and occipital lobes. These hemorrhages normally consisted of multiple repeated CLHs that frequently bled into the subarachnoid space. CAA-associated microvascular lesions may be the pathological factor underlying CLH.
Amyloid* ; Autopsy ; Cerebral Amyloid Angiopathy ; Diagnosis ; Hemorrhage* ; Necrosis ; Neurofibrillary Tangles ; Occipital Lobe ; Onions ; Plaque, Amyloid ; Rabeprazole ; Retrospective Studies ; Subarachnoid Space

OBJECTIVETo construct full-length hepatitis B core particles presenting preS1 aa 21-47 epitope and truncated core particles presenting preS1 aa 37-45 epitope on their surface and compare their antigenicity.

METHODSPreS1 aa21-47 epitope and aa 37-45 epitope were inserted respectively into full-length hepatitis B core (aa 1-183) and truncated HBcAg (aa 1-144), between the 78th (Asp) and 79th (Pro). The genes synthesized after the codon optimization were ligated to the pET43. 1a vector with the same cohesive terminal (NdeI and XhoI) and expressed in the E. coli expression system. The morphology of the proteins of interest were observed by electron microscope and characterized by ELISA and Western Blotting.

RESULTSThe morphology of the virus-like particles were confirmed by electron microscope. H2 were solid particles with a diameter of (31.61 +/- 1.27) nm, while H3 were hollow particles with a diameter of (28.46 +/- 1.16) nm. Statistical analysis showed that H2 is larger than H3 in the diameter (P < 0.01). The antigenicity of the inserted epitopes and carrier protein were identified by ELISA and Western Blotting.

CONCLUSIONChimeric hepatitis B core particles presenting the preS1 neutralizing epitopes on their surface have been expressed, purified and identified, which lays the foundation for its application in vaccine research.

Epitopes ; chemistry ; genetics ; immunology ; Hepatitis B ; immunology ; virology ; Hepatitis B Core Antigens ; chemistry ; genetics ; immunology ; Hepatitis B Surface Antigens ; chemistry ; genetics ; immunology ; Hepatitis B virus ; chemistry ; genetics ; immunology ; Humans ; Neutralization Tests ; Protein Precursors ; chemistry ; genetics ; immunology ; Recombinant Fusion Proteins ; chemistry ; genetics ; immunology