Comparison of different derivatisation for amino acids determination of foie gras by high performance liquid chromatography (HPLC).

1. In order to compare the difference between different derivatisations for amino acids determination of foie gras via, reversed phase high performance liquid chromatography (HPLC), O-phthalaldehyde and 9-fluorenyl-methyl chloroformate (OPA-FMOC group), phenylisothiocyanate (PITC group) and 6-Aminoquinolyl-N-hydrox-ysuccinimidyl Carbamate (AQC group) were applied to derivatisation reagent in this current experiment. The determination results of automatic amino acid analyser were applied, and 17 amino acids were detected by these three derivatisation methods.2. The running times of OPA-FMOC group, PITC group and AQC group were 18, 45 and 35 min, respectively. There was a large difference between the results of OPA-FMOC group and results from the automatic amino acid analyser, although the difference between the results from PITC and the automatic amino acid analyser was minimal.3. In conclusion, the running time of OPA-FMOC group was shorter than that of PITC group and AQC group; the accuracy of the former was better than the OPA-FMOC group and AQC group for the determination of amino acid of foie gras.

RXR agonists inhibit high glucose-induced upregulation of inflammation by suppressing activation of the NADPH oxidase-nuclear factor-κB pathway in human endothelial cells.

An inflammatory response induced by high glucose is a cause of endothelial dysfunction in diabetes and is an important contributing link to atherosclerosis. Diabetes is an independent risk factor of atherosclerosis and activation of retinoid X receptor (RXR) has been shown to exert anti-atherogenic effects. In the present study, we examined the effects of the RXR ligands 9-cis-retinoic acid (9-cis-RA) and SR11237 on high glucose-induced inflammation in human umbilical endothelial vein endothelial cells (HUVECs) and explored the potential mechanism. Our results showed that the inflammation induced by high-glucose in HUVECs was mainly mediated by the activation of nuclear factor-B (NF- κB). High glucose-induced expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were in comparison, significantly decreased by treatment with RXR. The effect of RXR agonists was mainly due to the inhibition of NF-κB activation. Using pharmacological inhibitors and siRNA, we confirmed that nicotinamide adenine dinucleotide phosphate (NADPH) oxidase was an upstream activator of NF-κB. Furthermore, RXR agonists significantly inhibited high glucose-induced activation of NADPH oxidase and significantly decreased the production of reactive oxygen species (ROS). To explore whether the rapid inhibitory effects of RXR agonists were in fact mediated by RXR, we examined the effect of RXR downregulation by RXR siRNA. Our results showed that RXR siRNA largely abrogated the effects of RXR agonists, suggesting the requirement of RXR expression. Therefore, we have shown that RXR is involved in the regulation of NADPH oxidase- NF-κB signal pathway, as the RXR ligands antagonized the inflammatory response in HUVECs induced by high glucose.

A phylogenetic study of dengue virus in urban Vietnam shows long-term persistence of endemic strains.

Dengue virus (DENV) causes repeated outbreaks of disease in endemic areas, with patterns of local transmission strongly influenced by seasonality, importation via human movement, immunity, and vector control efforts. An understanding of how each of these interacts to enable endemic transmission (continual circulation of local virus strains) is largely unknown. There are times of the year when no cases are reported, often for extended periods of time, perhaps wrongly implying the successful eradication of a local strain from that area. Individuals who presented at a clinic or hospital in four communes in Nha Trang, Vietnam, were initially tested for DENV antigen presence. Enrolled positive individuals then had their corresponding household members invited to participate, and those who enrolled were tested for DENV. The presence of viral nucleic acid in all samples was confirmed using quantitative polymerase chain reaction, and positive samples were then whole-genome sequenced using an amplicon and target enrichment library preparation techniques and Illumina MiSeq sequencing technology. Generated consensus genome sequences were then analysed using phylogenetic tree reconstruction to categorise sequences into clades with a common ancestor, enabling investigations of both viral clade persistence and introductions. Hypothetical introduction dates were additionally assessed using a molecular clock model that calculated the time to the most recent common ancestor (TMRCA). We obtained 511 DENV whole-genome sequences covering four serotypes and more than ten distinct viral clades. For five of these clades, we had sufficient data to show that the same viral lineage persisted for at least several months. We noted that some clades persisted longer than others during the sampling time, and by comparison with other published sequences from elsewhere in Vietnam and around the world, we saw that at least two different viral lineages were introduced into the population during the study period (April 2017-2019). Next, by inferring the TMRCA from the construction of molecular clock phylogenies, we predicted that two of the viral lineages had been present in the study population for over a decade. We observed five viral lineages co-circulating in Nha Trang from three DENV serotypes, with two likely to have remained as uninterrupted transmission chains for a decade. This suggests clade cryptic persistence in the area, even during periods of low reported incidence.

A new direct compression mechanism of structural transition in Poria cocos extract composite particles.

The structural transition to generate amorphous translucent grains in Poria cocos dry extract (PCE) composite particles was found and studied as a new direct compression mechanism. The pressure and displacement sensing techniques were used to obtained stress-strain profiles during compression. The Exponential function, Kawakita model, Shapiro model and Heckel model were used to analysis mechanical properties of powders. 12 parameters derived from compression models and powder physical properties were applied to partial least squares method (PLS) for analyzing powder compression mechanism. It was found that only the oven-dried PCE composite particles undergoes the structural transition and generate translucent grains scattered and embedded in tablet, and these tablets have excellent mechanical stability. The structural transition in plant dry extract as the PCE composite particles could be exploited to improve powder compression and tabletability.

Critical role of CXCL4 in the lung pathogenesis of influenza (H1N1) respiratory infection.

Annual epidemics and unexpected pandemics of influenza are threats to human health. Lung immune and inflammatory responses, such as those induced by respiratory infection influenza virus, determine the outcome of pulmonary pathogenesis. Platelet-derived chemokine (C-X-C motif) ligand 4 (CXCL4) has an immunoregulatory role in inflammatory diseases. Here we show that CXCL4 is associated with pulmonary influenza infection and has a critical role in protecting mice from fatal H1N1 virus respiratory infection. CXCL4 knockout resulted in diminished viral clearance from the lung and decreased lung inflammation during early infection but more severe lung pathology relative to wild-type mice during late infection. Additionally, CXCL4 deficiency decreased leukocyte accumulation in the infected lung with markedly decreased neutrophil infiltration into the lung during early infection and extensive leukocyte, especially lymphocyte accumulation at the late infection stage. Loss of CXCL4 did not affect the activation of adaptive immune T and B lymphocytes during the late stage of lung infection. Further study revealed that CXCL4 deficiency inhibited neutrophil recruitment to the infected mouse lung. Thus the above results identify CXCL4 as a vital immunoregulatory chemokine essential for protecting mice against influenza A virus infection, especially as it affects the development of lung injury and neutrophil mobilization to the inflamed lung.

Coadministration of VDR and RXR agonists synergistically alleviates atherosclerosis through inhibition of oxidative stress: An in vivo and in vitro study.

BACKGROUND AND AIMS: Diabetes contributes to atherosclerosis partially through induction of oxidative stress. Both vitamin D receptor (VDR) and retinoid X receptor (RXR) agonists exhibit anti-atherogenic effects. METHODS: We explored the effects of combination treatment with VDR and RXR agonists (represented by calcitriol and bexarotene, respectively) on atherosclerosis progression and the mechanisms involved, using a diabetes model of mice. The animals were intragastrically fed calcitriol (200 ng/kg, twice-a-week), bexarotene (10 mg/kg, once-daily) either alone or in combination for 12 weeks. RESULTS: VDR and RXR agonists delayed atherosclerosis progression independent of serum lipid and glucose levels, and significantly reduced the protein expression of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunit gp91phox and nuclear factor-kappa B (NF-κB) subunit p65, as well as plasma biomarkers of oxidative stress and inflammation. Combination therapy alleviated atherosclerosis and inhibited indexes of oxidative stress and inflammation to a greater extent than either monotherapy. In the in vitro study, naturally occurring VDR ligand 1α,25-dihydroxyvitamin D3 (1,25[OH]2D3) and RXR ligand 9-cis retinoic acid (9-cis-RA), both significantly inhibited high-glucose-induced endothelial cell apoptosis. Co-administration of VDR and RXR ligands produced synergistic protection against endothelial apoptosis by antagonizing the protein kinase C /NADPH oxidase/reactive oxygen species pathway. The inhibitory effects of 9-cis-RA on oxidative stress was attenuated when VDR was downregulated by VDR siRNA; however, downregulation of RXR by RXR siRNA imposed no influence on the effects of 1,25(OH)2D3. CONCLUSIONS: Combination treatment with VDR and RXR agonists synergistically alleviated diabetic atherosclerosis through inhibition of oxidative stress, and the preventive effects of RXR agonist may partially depend on VDR activation.

Quinazolines and 1,4-benzodiazepines. 82.1 5-Pyrimidyl-and 5-pyrazinylbenzodiazepines.

Analogues of bromazepam [7-bromo-1,3-dihydro-5(2-pyridyl)-2H-1,4-benzodiazepin-2-one, A], which is a clinically useful minor tranquilizer, have been prepared by replacing the 2-pyridyl group at position 5 with 4-pyrimidyl (5), 2-pyrazinyl (8), 2,5-dimethylpyrazin-3-yl (10), and 2-pyrimidyl (12) groups. Low to moderate CNS activities in both mice and cat were found for all the new compounds. For the screening procedures used, the 2-pyrimidyl-substituted derivatives were found to be the most active new analogues although none of the activities exceeded those observed for bromazepam.

A cone beam filtered backprojection (CB-FBP) reconstruction algorithm for a circle-plus-two-arc orbit.

The circle-plus-arc orbit possesses advantages over other "circle-plus" orbits for the application of x-ray cone beam (CB) volume CT in image-guided interventional procedures requiring intraoperative imaging, in which movement of the patient table is to be avoided. A CB circle-plus-two-arc orbit satisfying the data sufficiency condition and a filtered backprojection (FBP) algorithm to reconstruct longitudinally unbounded objects is presented here. In the circle suborbit, the algorithm employs Feldkamp's formula and another FBP implementation. In the arc suborbits, an FBP solution is obtained originating from Grangeat's formula, and the reconstruction computation is significantly reduced using a window function to exclude redundancy in Radon domain. The performance of the algorithm has been thoroughly evaluated through computer-simulated phantoms and preliminarily evaluated through experimental data, revealing that the algorithm can regionally reconstruct longitudinally unbounded objects exactly and efficiently, is insensitive to the variation of the angle sampling interval along the arc suborbits, and is robust over practical x-ray quantum noise. The algorithm's merits include: only 1D filtering is implemented even in a 3D reconstruction, only separable 2D interpolation is required to accomplish the CB backprojection, and the algorithm structure is appropriate for parallel computation.

Computer simulation of image intensifier-based computed tomography detector: vascular application.

This present study reports the results of a computer simulation whose aim was to predict the low-contrast imaging performance of which a conventional x-ray image intensifier with charge coupled device (CCD) camera would be capable if incorporated into a computed tomography (CT) volume imager. A vascular imaging task was modeled in our simulation. The effects of detector noise, x-ray exposure levels, analog-to-digital conversion (ADC) precision and residual levels of detected x-ray scatter were considered. The results of this simulation indicate that the low-contrast imaging performance of an image intensifier-based CT system was most limited by the CCD detector readout noise. Given this limitation the detection of greater than about 100,000 detected photons/pixel/projection gave marginal improvement in low-contrast resolution. At these exposures 12 bit ADC precision resulted in little additional image noise. The effects of detecting scattered x rays are twofold; decreasing the signal-to-noise ratio associated with our modeled artery and introducing a cupping artifact. Based on the results from the simulation, it appears that an image intensifier-based CT system is a feasible concept from a noise viewpoint, if the anticipated imaging task is intravenous angiography.

Cone beam volume CT image artifacts caused by defective cells in x-ray flat panel imagers and the artifact removal using a wavelet-analysis-based algorithm.

The application of x-ray flat panel imagers (FPIs) in cone beam volume CT (CBVCT) has attracted increasing attention. However, due to a deficient semiconductor array manufacturing process, defective cells unavoidably exist in x-ray FPIs. These defective cells cause their corresponding image pixels in a projection image to behave abnormally in signal gray level, and result in severe streak and ring artifacts in a CBVCT image reconstructed from the projection images. Since a three-dimensional (3-D) back-projection is involved in CBVCT, the formation of the streak and ring artifacts is different from that in the two-dimensional (2-D) fan beam CT. In this paper, a geometric analysis of the abnormality propagation in the 3D back-projection is presented, and the morphology of the streak and ring artifacts caused by the abnormality propagation is investigated through both computer simulation and phantom studies. In order to calibrate those artifacts, a 2D wavelet-analysis-based statistical approach to correct the abnormal pixels is proposed. The approach consists of three steps: (1) the location-invariant defective cells in an x-ray FPI are recognized by applying 2-D wavelet analysis on flat-field images, and a comprehensive defective cell template is acquired; (2) based upon the template, the abnormal signal gray level of the projection image pixels corresponding to the location-invariant defective cells is replaced with the interpolation of that of their normal neighbor pixels; (3) that corresponding to the isolated location-variant defective cells are corrected using a narrow-windowed median filter. The CBVCT images of a CT low-contrast phantom are employed to evaluate this proposed approach, showing that the streak and ring artifacts can be reliably eliminated. The novelty and merit of the approach are the incorporation of the wavelet analysis whose intrinsic multi-resolution analysis and localizability make the recognition algorithm robust under variable x-ray exposure levels between 30% and 70% of the dynamic range of an x-ray FPI.

Reconstruction of blood vessels from x-ray subtraction projections: limited angle geometry.

Several algorithms have been investigated for reconstructing blood vessels from a limited number of x-ray subtraction projections, distributed over a limited range of angles. Both computer simulations and an in vivo animal study were carried out. The best reconstruction performance was achieved using an algorithm that folded in two pieces of a priori knowledge of the vascular density distributions: (1) the object is dilute, consisting mainly of a void; and (2) the density distribution in the reconstructions is most likely to be non-negative. Both the signal-to-noise ratio (SNR) and the signal to out-of-focus blur were quantitated. Compared to tomosynthetic reconstruction (backprojection), the amount of residual blur from out-of-focus planes was significantly reduced with only a small penalty in diminished SNR. The combined effect resulted in significant qualitative image improvement for real arterial distributions as demonstrated in a canine arterial imaging example.

A cone-beam reconstruction algorithm for circle-plus-arc data-acquisition geometry.

In cone-beam computerized tomography (CT), projections acquired with the focal spot constrained on a planar orbit cannot provide a complete set of data to reconstruct the object function exactly. There are severe distortions in the reconstructed noncentral transverse planes when the cone angle is large. In this work, a new method is proposed which can obtain a complete set of data by acquiring cone-beam projections along a circle-plus-arc orbit. A reconstruction algorithm using this circle-plus-arc orbit is developed, based on the Radon transform and Grangeat's formula. This algorithm first transforms the cone-beam projection data of an object to the first derivative of the three-dimensional (3-D) Radon transform, using Grangeat's formula, and then reconstructs the object using the inverse Radon transform. In order to reduce interpolation errors, new rebinning equations have been derived accurately, which allows one-dimensional (1-D) interpolation to be used in the rebinning process instead of 3-D interpolation. A noise-free Defrise phantom and a Poisson noise-added Shepp-Logan phantom were simulated and reconstructed for algorithm validation. The results from the computer simulation indicate that the new cone-beam data-acquisition scheme can provide a complete set of projection data and the image reconstruction algorithm can achieve exact reconstruction. Potentially, the algorithm can be applied in practice for both a standard CT gantry-based volume tomographic imaging system and a C-arm-based cone-beam tomographic imaging system, with little mechanical modification required.

Flat panel detector-based cone-beam volume CT angiography imaging: system evaluation.

Preliminary evaluation of recently developed large-area flat panel detectors (FPDs) indicates that FPDs have some potential advantages: compactness, absence of geometric distortion and veiling glare with the benefits of high resolution, high detective quantum efficiency (DQE), high frame rate and high dynamic range, small image lag (< 1%), and excellent linearity (approximately 1%). The advantages of the new FPD make it a promising candidate for cone-beam volume computed tomography (CT) angiography (CBVCTA) imaging. The purpose of this study is to characterize a prototype FPD-based imaging system for CBVCTA applications. A prototype FPD-based CBVCTA imaging system has been designed and constructed around a modified GE 8800 CT scanner. This system is evaluated for a CBVCTA imaging task in the head and neck using four phantoms and a frozen rat. The system is first characterized in terms of linearity and dynamic range of the detector. Then, the optimal selection of kVps for CBVCTA is determined and the effect of image lag and scatter on the image quality of the CBVCTA system is evaluated. Next, low-contrast resolution and high-contrast spatial resolution are measured. Finally, the example reconstruction images of a frozen rat are presented. The results indicate that the FPD-based CBVCT can achieve 2.75-lp/mm spatial resolution at 0% modulation transfer function (MTF) and provide more than enough low-contrast resolution for intravenous CBVCTA imaging in the head and neck with clinically acceptable entrance exposure level. The results also suggest that to use an FPD for large cone-angle applications, such as body angiography, further investigations are required.

Radioimmunoassay of the anticonvulsant agent clonazepam.

A simple and specific radioimmunoassay was developed for the determination of the anticonvulsant agent clonazepam directly in plasma without extraction. Antibodies to clonazepam were produced in rabbits after immunization with an immunogen prepared by covalently linking the 3-hemisuccinyloxy derivative of clonazepam to bovine serum albumin. When employing 3H-clonazepam as the tracer, the radioimmunoassay has a limit of sensitivity of 5 ng/ml using a 0.1-ml sample of plasma. The antibodies exhibited a high degree of specificity for clonazepam; no cross-reactivity was observed with its 7-amino and 7-acetylamino metabolites nor with a number of other widely prescribed anticonvulsant agents that might be administered in conjuction with clonazepam. Satisfactory agreement was obtained for the plasma levels of clonazepam in humans when samples were assayed by the radioimmunoassay and an established electron-capture GC technique. By virtue ot its simplicity, the radioimmunoassay offers a distinct advantage to the clinician for monitoring plasma clonazepam levels and the compliance of patients undergoing anticonvulsant therapy with the drug.

Quinazolines and 1,4-benzodiazepines. LXXXIX: Haptens useful in benzodiazepine immunoassay development.

The syntheses of some 1,4-benzodiazepines potentially useful as haptens are reported. These compounds are related to chlordiazepoxide, diazepam, nitrazepam, clonazepam, and some of their metabolites. The chemistry reported here is intended to support specific immunoassay development for these drugs.

Image intensifier-based computed tomography volume scanner for angiography.

RATIONALE AND OBJECTIVES: A prototype volume computed tomography (CT) system for use in angiography was designed, constructed, and tested. The system consisted of a fixed X-ray tube, a conventional image intensifier (II) coupled to a charge-coupled device camera, and a computer-controlled turntable on which phantoms were placed. We wanted to predict, through phantom studies, the imaging performance of an II-based volume CT for direct three-dimensional (3D) reconstruction of vascular structures. METHODS: To explore the imaging performance of the system for reconstructing a vascular structure, two sets of projection images of a vascular phantom, acquired over 250 projection angles with two different-sized IIs, were digitized and used for a direct 3D conebeam reconstruction. The signal-to-noise ratio (SNR) of each reconstructed image was measured. From these measurements, image quality was accessed as a function of the number of reconstructions averaged and the different orientations. The spatial resolution limits of the system were measured from the 3D reconstructed images of a specially designed resolution phantom for different orientations and locations. RESULTS: The measured SNRs of all direct 3D reconstruction images were reasonably good, and back-ground noise levels measured from 3D reconstruction images were almost 30 Hounsfield units. The measured spatial resolution of the system was 0.5 line pairs per millimeter. However, spatial resolution was reduced around the edge of the II to nearly half that measured in the central area of the field of view. CONCLUSION: An II-based volume CT scanner can produce direct 3D reconstructions of vascular structures with good image quality for intraarterial angiography.

MR volumetric analysis of the human basal ganglia: normative data.

RATIONALE AND OBJECTIVES: The authors undertook this study to identify a precise, semiautomated, reproducible magnetic resonance (MR) imaging technique for measuring the basal ganglia, to establish normative volumetric data, and to verify the presence of previously reported asymmetries. MATERIALS AND METHODS: Twenty-eight healthy adults underwent cranial MR examination. The volumes of the various components of the basal ganglia were measured by means of a combination of thresholding and manual tracing techniques performed with specialized software. The validity of these measurements was assessed by fashioning, imaging, and measuring a practical basal ganglia phantom. Measurement accuracy was also established by means of inter- and intrarater reliability indexes. Normalized volumes were statistically analyzed with analysis of variance and paired t tests. RESULTS: The absolute values of the various components of the basal ganglia varied widely even though the volumes were normalized to differences in intracranial volume. The right caudate nucleus volume was significantly (P < .000001) larger than the left in both men and women and in both right-handed and non-right-handed subjects. This asymmetry led to an increase in the overall volume of the basal ganglia on the right. CONCLUSION: The authors have defined a precise, reproducible technique for measuring various components of the basal ganglia and have established normative data. The basal ganglia, similar to other brain structures, exhibit hemispheric lateralization.

Neamine induces neuroprotection after acute ischemic stroke in type one diabetic rats.

INTRODUCTION: Angiogenin is a member of the ribonuclease superfamily and promotes degradation of the basement membrane and the extracellular matrix. After stroke in type one diabetes (T1DM) rats, Angiogenin is significantly increased and the Angiogenin is inversely correlated with functional outcome. Neamine, an aminoglycoside antibiotic, blocks nuclear translocation of Angiogenin, thereby abolishing the biological activity of Angiogenin. In this study, we therefore investigated the effect and underlying protective mechanisms of Neamine treatment of stroke in T1DM. METHODS: T1DM was induced in male Wistar rats by streptozotocin (60mg/kg, ip), and T1DM rats were subjected to embolic middle cerebral artery occlusion (MCAo). Neamine (10mg/kg ip) was administered at 2, 24 and 48h after the induction of embolic MCAo. A battery of functional outcome tests was performed. Blood-brain barrier (BBB) leakage, and lesion volume were evaluated and immunostaining, and Western blot were performed. RESULTS: Neamine treatment of stroke in T1DM rats significantly decreased BBB leakage and lesion volume as well as improved functional outcome compared to T1DM-control. Neamine also significantly decreased apoptosis and cleaved caspase-3 in the ischemic brain. Using immunostaining, we found that Neamine treatment significantly decreased nuclear Angiogenin, nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) activity, advanced glycation endproducts receptor (RAGE) number, the positive area of toll-like receptor 4 (TLR4) and increased Angeopoietin-1 expression compared to T1DM-MCAo control rats. Western blot results are consistent with the immunostaining. CONCLUSION: Neamine treatment of stroke is neuroprotective in T1DM rats. Inhibition of neuroinflammatory factor expression and decrease of BBB leakage may contribute to Neamine-induced neuroprotective effects after stroke in T1DM rats.

Tissue plasminogen activator treatment of stroke in type-1 diabetes rats.

BACKGROUND AND PURPOSE: Diabetes mellitus (DM) is a major stroke risk factor and is associated with poor recovery compared with nondiabetic stroke patients. In the present study, we investigated the effects of tissue plasminogen activator (tPA) treatment of stroke in diabetic and non-diabetic rats. METHODS: Type-1 diabetes (T1DM) was induced by injection of streptozotocin. Non-T1DM and T1DM rats were subjected to embolic middle cerebral artery occlusion (MCAo) and treated with or without tPA 2h after MCAo. Functional outcomes and immunostaining for advanced glycation endproducts receptor (RAGE), matrix metalloproteinase-9 (MMP-9) and toll-like receptor 4 (TLR4) and Western blotting were performed. RESULTS: tPA treatment of WT-MCAo rats significantly improved the functional outcome and reduced the lesion volume compared with non-treatment WT-MCAo rats (p<0.05). There was no significant difference between treatment with or without tPA in the WT-MCAo group in brain hemorrhage, BBB leakage and expression of inflammatory mediators, RAGE, MMP-9 and TLR4. However, tPA treatment in T1DM-MCAo rats (T1DM-MCAo+tPA) significantly enlarged brain hemorrhage, augmented BBB leakage, and failed to decrease lesion volume and improve functional outcome after stroke compared to T1DM-MCAo control. tPA treatment also significantly increased the expression of RAGE, MMP-9 and TLR4 in the ischemic brain in T1DM-MCAo rats compared with T1DM-MCAo control rats (p<0.05). Brain hemorrhage was significantly correlated with functional deficit and RAGE and TLR4 expression, respectively. CONCLUSIONS: Treatment of stroke with tPA increased brain hemorrhage, BBB leakage and failed to improve functional outcome in T1DM rats. The increased inflammatory response may contribute to the failed neuroprotective effects of tPA treatment in T1DM rats.

Combination treatment of stroke with sub-therapeutic doses of Simvastatin and human umbilical cord blood cells enhances vascular remodeling and improves functional outcome.

Human umbilical cord blood cells (HUCBCs) have been employed as a restorative treatment for experimental stroke. In this study, we investigated whether transplantation of sub-therapeutic doses of HUCBCs and Simvastatin enhances cerebral vascular remodeling after stroke. Adult male Wistar rats (n=34) were subjected to transient middle cerebral artery occlusion (MCAo) and treated with: phosphate-buffered solution (PBS, gavaged daily for 7 days); Simvastatin (0.5mg/kg, gavaged daily for 7 days); HUCBCs (1×10(6), injected once via tail vein); and combination Simvasatin with HUCBCs, starting at 24h after MCAo. There was no significant difference between Simvastatin- or HUCBC-monotherapy and MCAo-alone group. Combination treatment 24h post-stroke significantly increased the perimeter of von Willebrand factor (vWF)-positive vessels, the diameter and density of alpha smooth muscle actin (αSMA)-positive arteries, and the percentage of 5-bromodeoxyuridine (BrdU)-positive endothelial cells (ECs) in the ischemic boundary zone (IBZ) compared with MCAo-alone or HUCBC-monotherapy 14 days after MCAo (p<0.05, n=8/group); Combination treatment significantly increased the densities of vWF-vessels and αSMA-arteries as well as the densities of BrdU-ECs and BrdU-positive smooth muscle cells (SMCs) in vascular walls in the IBZ compared with Simvastatin-monotherapy. Moreover, the increased BrdU-ECs and BrdU-SMCs were significantly correlated with neurological functional outcome 14 days after MCAo. Combination treatment also significantly increased the expression of Angiopoietin-1 (Ang1), Tie2 and Occludin in the IBZ (p<0.05, n=8/group). The in vitro experiments showed that combination treatment and Ang1 significantly increased capillary-like tube formation and arterial cell migration; anti-Ang1 significantly reduced combination treatment-induced tube-formation and artery cell migration (p<0.05, n=6/group). These findings indicated that a combination of sub-therapeutic doses of Simvastatin and HUCBCs treatment of stroke increases Ang1/Tie2 and Occludin expression in the ischemic brain, amplifies endogenous angiogenesis and arteriogenesis, and enhances vascular remodeling which in concert may contribute to functional outcome after stroke.