Plasminogen deficiency causes reduced corticospinal axonal plasticity and functional recovery after stroke in mice.

Tissue plasminogen activator (tPA) has been implicated in neurite outgrowth and neurological recovery post stroke. tPA converts the zymogen plasminogen (Plg) into plasmin. In this study, using plasminogen knockout (Plg-/-) mice and their Plg-native littermates (Plg+/+), we investigated the role of Plg in axonal remodeling and neurological recovery after stroke. Plg+/+ and Plg-/- mice (n = 10/group) were subjected to permanent intraluminal monofilament middle cerebral artery occlusion (MCAo). A foot-fault test and a single pellet reaching test were performed prior to and on day 3 after stroke, and weekly thereafter to monitor functional deficit and recovery. Biotinylated dextran amine (BDA) was injected into the left motor cortex to anterogradely label the corticospinal tract (CST). Animals were euthanized 4 weeks after stroke. Neurite outgrowth was also measured in primary cultured cortical neurons harvested from Plg+/+ and Plg-/- embryos. In Plg+/+ mice, the motor functional deficiency after stroke progressively recovered with time. In contrast, recovery in Plg-/- mice was significantly impaired compared to Plg+/+ mice (p0.82, p<0.01). Plg-/- neurons exhibited significantly reduced neurite outgrowth. Our data suggest that plasminogen-dependent proteolysis has a beneficial effect during neurological recovery after stroke, at least in part, by promoting axonal remodeling in the denervated spinal cord

Failure Analysis of Casing in Shale Oil Wells under Multistage Fracturing Conditions

During the multistage fracturing in shale oil and gas wells with tieback and liner, one of the major challenges is the wellbore temperature variation due to the high-rate fracturing. In such a case, the axial shrinkage trend of the casing string could be caused due to the sudden drop in temperature, but the actual axial length of the casing string would not change due to the cement constraints. Therefore, this could lead to cementation damage between the casing and cement due to excessive load from the casing string. A wellbore seal that is out of control often leads to irreversible consequences, even well abandonment. In order to study the mechanism of casing deformation in shale oil and gas wells with tieback and liner quantitatively, in this paper, take LS1 well (a typical shale oil and gas well with tieback and liner, and casing deformation is caused) for example, the transient changes of temperature and pressure in the whole wellbore during multistage fracturing are studied. Moreover, the cementing strength test of the interface between casing and cement is also tested. Then, the testing results are carried out and extended to model the finite element (FE) model with the whole vertical section casing string with tieback and liner. The model is used to simulate the internal force changes under fracturing conditions with different stages of fracturing. Meanwhile, the casing deformation mechanism in LS1 well is analyzed and studied in detail. Our simulation results indicated the failure process and mechanism of cementation between casing and cement in shale oil and gas wells with tieback and liner. Our work can provide a detailed theoretical reference and a basis for field application

Efficacy of single and multiple injections of human umbilical tissue-derived cells following experimental stroke in rats.

INTRODUCTION: Human umbilical tissue-derived cells (hUTC) are a promising source of cells for regenerative treatment of stroke. In this study, we tested the efficacy of hUTC in experimental stroke and whether multiple injections of hUTC provide additional therapeutic benefits as compared to a single injection. METHODS: Adult male Wistar rats were subjected to 2 hours of middle cerebral artery occlusion (MCAo), and randomly selected animals were injected (i.v) with 3×10⁶ hUTC or with vehicle control (at day: 1, 1&3 or 1&7 after MCAo, n = 8-9/group). A battery of functional outcome tests was performed at days 1, 7, 14, 21, 28, 35, 42, 49, 56 and 63 after MCAo. Rats were sacrificed at 63 days after MCAo and lesion volumes were measured. To investigate the underlying mechanism of hUTC treatment of stroke, Von Willebrand Factor (vWF), and Synaptophysin immunostaining were performed. RESULTS: All hUTC treated groups, single or multiple injections, had better functional recovery compared to control (p<0.01). There was no statistically significant difference between a single and multiple injections of hUTC (p = 0.23) or between different multiple injections groups (p>0.07) in functional outcome. All hUTC treatment groups showed significant increases in Synaptophysin, vascular density and perimeter compared to the control group (p<0.05). There was no statistically significant difference between a single and multiple injections of hUTC or between the two groups of multiple injections in all immunohistochemical measurements (p>0.1). CONCLUSION: hUTC treatment significantly improves long term functional outcome after stroke and promotes vascular density and synaptic plasticity. At the proscribed doses, multiple injections of hUTC were not superior to single injection therapy in both functional outcome and histological assessments

Ischemic lesion volume.

<p>A: A representative coronal section stained with hematoxylin and eosin shows ischemic infarct area. B: Quantitative data show there was no difference on the lesion volume between the Plg^+/+ and Plg^-/- mice at 28 days after stroke (n = 10/group). Scale bar = 1 mm.</p

Behavioral outcome after MCAo.

<p>A: The foot-fault test measures the accuracy of forepaw placement on a non-equidistant grid as the percentage of foot-faults of the left forepaw to total steps. B: The single pellet reaching test measures the ability of skilled forepaw use. Animals were trained to use their left forepaw to extract food pellets through a vertical slot of the front wall. The number of the left forepaw extensions through the slot and the number of pellets extracted were counted. Performance was defined by the success rate as the percentage of pellet number extracted per left forepaw attempts. After stroke, significant behavioral deficits were evident in both tests. The Plg^+/+ mice showed a progressive improvement with time, while the Plg^-/- mice exhibited a significant delayed recovery compared with the Plg^+/+ mice (n = 10/group, *p<0.01, **p<0.001).</p

Neurite outgrowth in primary cultured cortical neurons harvested from Plg^+/+ and Plg^-/- embryos.

<p>A representative image shows the neurons identified with immunofluorescent staining for beta tubulin (Tuj1, A). Compared to Plg^+/+ neurons, the Plg^-/- neurons exhibited a significantly reduced neurite outgrowth measured in the percentage of neurite positive neurons (B), number of neurite (C) and neurite length (D, n = 100, *p<0.01).</p