Aim Vascular endothelial cell growth inhibitor(VEGI) is a recently discovered novel member of the TNF superfamily,which is expressed predominantly in endothelial cells.As an endothelial cell-specific negative regulator of angiogenesis,the relationship between structure and function of VEGI is not understood at present.Methods In order to explore the functional key amino acids of VEGI,four mutants of VEGI(E45→R,G47→A,Y111→F,Y111→T) were construced by site-directed mutagenesis,and recombinant proteins were generated from E.coli.Four mutant proteins behaved similar to the wild type VEGI in various physico-chemical assays.The proliferation of HUVEC and chick choriallantic membrane assay were performed to study the activity of four mutants.Results The mutant E45→R significantly decreased the biological activity,and the mutant G47→A caused a slight drop on activity,but the mutants Y111→F,Y111→T almost completely abolished biological activity.Conclusion It suggests that Y111 is an important residue in biological activity,which may play a direct role in receptor recognition.Moreover,the tyrosine ring and hydroxy group of the amino acid are important determinant of biological activity.Additionally,E45 also plays an important role in biological activity of VEGI.

A new cool-tip radiofrequency (RF) ablation therapeutic instrument based on impedance control algorithm is introduced in this paper. The equipment is composed of hardware system and software system. The RF power output and real time data acquisition are completed by the hardware system, while the software is used mainly to finish the control of the ablation range, the core of which is impedance control algorithm, and it also used to complete the display of the real time data in the course of the experiment. The impedance algorithm has solved the problem of impedance increased rapidly during the RF ablation, which has also expanded the scope of ablation. The pig liver experiments showed that the impedance control algorithm had strong adaptability. It also obtained a result of ablation range up to 3.5-4.5 cm single needle. It has the high clinical practical value of one-time inactivation of 3-5 cm tumor.
Algorithms ; Animals ; Catheter Ablation ; Electric Impedance ; Humans ; Liver ; Needles ; Software ; Swine

Objective To analyze the role of tissue factor (TF) regulating the invasive ability of human colorectal carcinoma cells (HT-29). Method The plasmids pcDNA3.1/Zeo bearing either sense or antisense TFcDNA were transfected into HT-29 cells by lipofactamine 2000. TF proteins in transfectants and its parental cells were detected by Western blot. Matrigel invasion assays and nude mice inoculation assays were used to show the invasive ability of cancer cells in vitro and in vivo respectively. Microvessel density of the implanted tumors was counted. Result HT-29 cells with sense-TFcDNA transfection upregulated TF expression and invasive ability compared with the parental cells, and tumor tissues after sense transfection were of greater microvessel density than the parental cells. HT-29 cells transfected with antisense-TFcDNA had a downregulated TF expression and are of less microvessel density. Conclusion Positive TF expression increases the invasive ability of HT-29 cell in vitro and in vivo.

Objective To investigate the expression of T cell immunoglobulin-and mucin-domaincontaining molecule-3 (Tim-3) in peripheral CD8 +T cells and its significance in patients with chronic hepatitis B (CHB).Methods Fifty-eight CHB patients and 16 healthy controls were enrolled.Tim-3 expression in CDs + T cells was detected by flow cytometry,and quantities of IFNγ-producing HBV-specific cytotoxic T lymphocytes (CTLs) in HLA-A2 positive subjects were detected by enzyme-linked immunosorbent spot (ELISPOT) test before and after the blockade of Tim-3/Tim-3L pathway.Paired t test was performed to compare the quantities of CTLs before and after the blockade,and nonparametric Spearman correlation analysis was performed to explore the correlation in quantitive data.Results Tim-3 expression in CHB patients was (14.2 ± 8.98 )％,which was higher than that of healthy controls (4.80 ± 2.92)％,and the difference was of statistical significance (x2 =92.48,P ＜ 0.05 ) Tim-3 expressions in 16 severe CHB patients and 42 mild CHB patients were ( 19.54 ± 10.95) ％ and (9.58 ± 7.30) ％,respectively,and the difference was statistically significant (x2 =77.24,P ＜ 0.05 ). Before the blockade of Tim-3/Tim-3L pathway,IFNγ-producing HBV-specific CTLs were 7.27 ± 3.14,and it increased to 19.62 ± 4.97 after the blockade ( t =2.95,P ＜ 0.05 ).Conclusion The upregulation of Tim-3 on peripheral CD8 + T cells may inhibit HBV-specific CTLs,and the blockade of Tim-3 pathway can enhance the proliferation of IFNγ-producing HBV-specific CTLs,thus can enhance antiviral effect.

This study investigated the effect of arctigenin (Arc) on the cell activation, cytokines expression, proliferation, and cell-cycle distribution of mouse T lymphocytes. Mouse lymphocytes were prepared from lymph node and treated with Phorbol-12-myristate-13-acetate (PMA)/Ionimycin (Ion) and/or Arc. CD69, CD25, cytokines, proliferation and cell cycle were assayed by flow cytometry. The results showed that, at concentrations of less than 1.00 micromol x L(-1), Arc expressed non-obvious cell damage to cultured lymphocytes, however, it could significantly down-regulate the expression of CD69 and CD25, as well as TNF-alpha, IFN-gamma, IL-2, IL-4, IL-6 and IL-10 on PMA/Ion stimulated lymphocytes. At the same time, Arc could also inhibit the proliferation of PMA/Ion-activated lymphocytes and exhibited lymphocyte G 0/G1 phase cycle arrest. These results suggest that Arc possesses significant anti-inflammatory effects that may be mediated through the regulation of cell activation, cytokines expression and cell proliferation.
Animals ; Anti-Inflammatory Agents ; isolation & purification ; pharmacology ; Antigens, CD ; metabolism ; Antigens, Differentiation, T-Lymphocyte ; metabolism ; Arctium ; chemistry ; Cell Cycle Checkpoints ; drug effects ; Cell Proliferation ; drug effects ; Cytokines ; metabolism ; Female ; Furans ; isolation & purification ; pharmacology ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Ionomycin ; pharmacology ; Lectins, C-Type ; metabolism ; Lignans ; isolation & purification ; pharmacology ; Lymphocyte Activation ; drug effects ; Mice ; Mice, Inbred BALB C ; Plants, Medicinal ; chemistry ; T-Lymphocytes ; cytology ; drug effects ; immunology ; Tetradecanoylphorbol Acetate ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo study the plasma protein binding rate of methyl protodioscin.

METHODThe ultrafiltration was employed to determine the plasma protein binding rate of methyl protodioscin. The plasma concentrations of methyl protodioscin were measured by HPLC-MS-MS.

RESULTThe plasma protein binding rate of methyl protodioscin with rat plasma at the concentration of 20.0, 100 and 200 microg x mL(-1) were (94.6 +/- 0.16)%, (91.6 +/- 0.35)% and (86.10 +/- 0.60)%, respectively, while the plasma protein binding rate of methyl protodioscin with normal human plasma at the above concentrations were (82.11 +/- 5.12)%, (84.54 +/- 0.32)% and (88.52 +/- 1.02)%, respectively.

CONCLUSIONThe binding rate of methyl protodioscin with plasma protein is high.

Animals ; Antineoplastic Agents ; metabolism ; Blood Proteins ; metabolism ; Calibration ; Chromatography, High Pressure Liquid ; Diosgenin ; analogs & derivatives ; metabolism ; Female ; Humans ; Male ; Protein Binding ; Rats ; Saponins ; metabolism ; Sensitivity and Specificity ; Tandem Mass Spectrometry ; Ultrafiltration

BACKGROUNDFor patients with obstructive sleep apnea-hypopnea syndrome (OSAHS) and type 2 diabetes mellitus (T2DM), the night sleep interruption and intermittent hypoxia due to apnea or hypopnea may induce glycemic excursions and reduce insulin sensitivity. This study aimed to investigate the effect of continuous positive airway pressure (CPAP) therapy in patients with OSAHS and T2DM.

METHODSContinuous glucose monitoring system (CGMS) was used in 40 patients with T2DM and newly diagnosed OSAHS. The measurements were repeated after 30 days of CPAP treatment. Subsequently, insulin sensitivity and glycohemoglobin (HbA1c) were measured and compared to the pretreatment data.

RESULTSAfter CPAP therapy, the CGMS indicators showed that the 24-h mean blood glucose (MBG) and the night time MBG were significantly reduced (P < 0.05 and P = 0.03, respectively). The mean ambulatory glucose excursions (MAGEs) and the mean of daily differences were also significantly reduced (P < 0.05 and P = 0.002, respectively) compared to pretreatment levels. During the night, MAGE also significantly decreased (P = 0.049). The differences between the highest and lowest levels of blood glucose over 24 h and during the night were significantly lower than prior to CPAP treatment (P < 0.05 and P = 0.024, respectively). The 24 h and night time durations of high blood glucose (>7.8 mmol/L and > 11.1 mmol/L) decreased (P < 0.05 and P < 0.05, respectively) after the treatment. In addition, HbA1c levels were also lower than those before treatment (P < 0.05), and the homeostasis model assessment index of insulin resistance was also significantly lower than before CPAP treatment (P = 0.034).

CONCLUSIONSCPAP therapy may have a beneficial effect on improving not only blood glucose but also upon insulin sensitivity in T2DM patients with OSAHS. This suggests that CPAP may be an effective treatment for T2DM in addition to intensive diabetes management.

Adult ; Blood Glucose ; physiology ; Continuous Positive Airway Pressure ; methods ; Diabetes Mellitus, Type 2 ; therapy ; Female ; Humans ; Insulin Resistance ; physiology ; Male ; Middle Aged ; Sleep Apnea, Obstructive ; therapy ; Treatment Outcome

OBJECTIVETo observe the biocompatibility of new biomaterials porous calcium phosphate (CPC) and ectopic bone formation of CPC with bone marrow stromal cells (BMSCs).

METHODSThe BMSCs were cultured from Beagle dog and combined with the porous CPC with the best concentration after transfect green fluorescent protein (GFP). The adhesion and growth of BMSCs on CPC were observed under inversion, fluorescence and scanning electron microscopy. The ectopic bone formation were observed at the 8th week after CPC and BMSCs were implanted subcutaneously into nude mice.

RESULTSWhen BMSCs with CPC were cultured at the 1st day, cells were climbing out from CPC with normal morphology. At the 7th day cells can be seen protruding pseudopods, secretion of matrix. Bone formation could be seen histomorphologically at the 8th week.

CONCLUSIONPorous CPC has good biocompatibility and is an ideal scaffold material for bone tissue engineering.

Animals ; Biocompatible Materials ; Bone Cements ; Bone and Bones ; Calcium Phosphates ; Dental Cementum ; Dogs ; Mesenchymal Stromal Cells ; Mice ; Mice, Nude ; Tissue Engineering

OBJECTIVETo investigate the clinical characteristics and the corneal morphological abnormalities in Fleck corneal dystrophy.

METHODSEighteen eyes with Fleck corneal dystrophy of 9 patients from two unrelated families were examined by slit-lamp biomicroscopy, Cochet-Bonnet esthesiometer and in vivo confocal microscopy. The corneal cells, nerves and stromal deposits were analyzed quantitatively with NAVIS software.

RESULTSlit-lamp biomicroscopy of the Fleck corneal dystrophy revealed that bilateral small grey-white fleck-like or wreath-like opacities were scattered in all layer of the corneal stroma from the center to the periphery, the intervening stroma between the lesions was clear. In vivo confocal microscopy identified that the opacities appeared as doughnut-like or nephroid-like deposits approximately 70.6 μm × 110.3 μm in size, (1.6 ± 0.4)/frame in density and involved about (438.4 ± 22.0) μm of the corneal stroma, with hyper reflective dot-like intracellular particles,measuring 2 to 18 μm in diameter. In two eyes of 1 patient associated with decreased corneal sensation, confocal images showed that abnormal hyper reflective deposits involved the Bowman's layer and the branch and density of the subbasal nerve was reduced. The other eyes with normal corneal sensation appeared normal in the morphology and the density of the corneal nerves.

CONCLUSIONFleck corneal dystrophy has typical clinical characteristics. In vivo confocal microscopy allows to study the morphological changes of the cornea at cellular level, which is valuable for the clinical diagnosis and evaluation of the corneal dystrophy. Decreased corneal sensitivity in Fleck corneal dystrophy is probably caused by corneal stromal deposits involved in the Bowman's layer.

Adolescent ; Adult ; Aged ; Cornea ; pathology ; Corneal Dystrophies, Hereditary ; pathology ; Female ; Humans ; Male ; Microscopy, Confocal ; Middle Aged ; Young Adult

OBJECTIVETo evaluate the safety and efficacy and summarize the initial experience of transcatheter aortic valve implantation (TAVI) for treating patients with severe aortic stenosis.

METHODSFrom October 2010 to May 2011, TAVI using 18 F Corevalve system was applied in 3 patients with severe calcified aortic valve stenosis at high risk for surgery. The efficacy and complications of the procedure were analyzed and the procedure experiences were summarized.

RESULTSTAVI procedure was successful in all 3 cases. The mean operation time was (109.0 ± 22.6) minutes and X-ray exposure time was (24.0 ± 9.5) minutes. The peak pressure gradients after surgery were significantly reduced [from (84 ± 15) mm Hg (1 mm Hg = 0.133 kPa) to (6 ± 3) mm Hg]. A trivial to mild paravalvular leak was observed in all patients post procedure. Case 1 was free from perioperative complications. Case 2 experienced a transient complete left bundle branch block. Case 3 developed 3 degree atrioventricular block and implanted with a permanent cardiac pacemaker, cardiac tamponade which was relieved through conservative treatment, including pericardial puncture and drainage and acute kidney injury.

CONCLUSIONSOur initial experience showed that TAVI using the 18 F Corevalve system is safe and effective for patients with severe calcified aortic valve stenosis at high-risk for surgery, though the procedure may cause some complications. Strict patient selection and proficient surgical techniques may reduce the incidence of complications.

Aged ; Aged, 80 and over ; Aortic Valve ; surgery ; Aortic Valve Stenosis ; surgery ; Cardiac Catheterization ; Female ; Heart Valve Prosthesis Implantation ; methods ; Humans ; Male ; Treatment Outcome