Vigour testing for the rice seed with computer vision-based techniques.

Rice is the staple food for approximately half of the world's population. Seed vigour has a crucial impact on the yield, which can be evaluated by germination rate, vigor index and etc. Existing seed vigour testing methods heavily rely on manual inspections that are destructive, time-consuming, and labor-intensive. To address the drawbacks of existing rice seed vigour testing, we proposed a multispectral image-based non-destructive seed germination testing approach. Specifically, we collected multispectral data in 19 wavebands for six rice varieties. Furthermore, we designed an end-to-end pipeline, denoted as MsiFormer (MisFormer cod3e will be available at https://github.com/LiaoYun0x0/MisFormer) by integrating a Yolo-based object detector (trained Yolo v5) and a vision transformer-based vigour testing model, which effectively improved the automation and efficiency of existing techniques. In order to objectively evaluate the performance of the proposed method in this paper, we conduct a comparison between MisFormer and other 3 deep learning methods. The results showed that, MisFormer performed much better with the accuracy of 94.17%, which was 2.5%-18.34% higher than the other 3 deep learning methods. Besides MsiFormer, possibilities of CIELab mediated image analysis of TTC (tetrazolium chloride) staining in rice seed viability and nCDA (normalized canonical discriminant analysis) in rice seed vigour were also discussed, where CIELab L* of TTC staining were negatively correlated with vigor index and germination rate, with Pearson's correlation coefficient of -0.9874, -0.9802 respectively, and CIELab A* of TTC staining were and positively correlated with vigor index and germination rate, with Pearson's correlation coefficient of 0.9624, 0.9544 respectively, and CIELab A* of nCDA had Pearson's correlation coefficient of -0.8866 and -0.9340 with vigor index and germination rate, respectively. Besides testing methods, vigour results within and among variety(ies) showed that, there were great variations among the 6 rice varieties, and mean coefficient of variation (CV) of vigor index of individual seed within a variety reached 64.87%, revealing the high risk of conventional methods in random sampling. Vigour variations had close relationship with wavelengths of 780 nm-970 nm, indicating their value in future research.

The superiority and feasibility of 2,3,5-triphenyltetrazolium chloride-stained brain tissues for molecular biology experiments based on microglial properties.

BACKGROUND: TTC (2,3,5-triphenyltetrazolium chloride) staining is the most commonly used method in identifying and assessing cerebral infarct volumes in the transient middle cerebral artery occlusion model. Given that microglia exhibit different morphologies in different regions after ischemic stroke, we demonstrate the superiority and necessity of using TTC-stained brain tissue to analyze the expression of various proteins or genes in different regions based on microglia character. METHODS: We compared brain tissue (left for 10 min on ice) from the improved TTC staining method with penumbra from the traditional sampling method. We identified the feasibility and necessity of the improved staining method using real time (RT)-PCR, Western blot, and immunofluorescence analysis. RESULTS: There was no protein and RNA degradation in the TTC-stained brain tissue group. However, the TREM2 specifically expressed on the microglia showed a significant difference between two groups in the penumbra region. CONCLUSIONS: TTC-stained brain tissue can be used for molecular biology experiments without any restrictions. In addition, TTC-stained brain tissue shows greater superiority due to its precise positioning.

Tectonic infarct analysis: A computational tool for automated whole-brain infarct analysis from TTC-stained tissue.

Background: Infarct volume measured from 2,3,5-triphenyltetrazolium chloride (TTC)-stained brain slices is critical to in vivo stroke models. In this study, we developed an interactive, tunable, software that automatically computes whole-brain infarct metrics from serial TTC-stained brain sections. Methods: Three rat ischemic stroke cohorts were used in this study (Total n = 91 rats; Cohort 1 n = 21, Cohort 2 n = 40, Cohort 3 n = 30). For each, brains were serially-sliced, stained with TTC and scanned on both anterior and posterior sides. Ground truth annotation and infarct morphometric analysis (e.g., brain-Vbrain, infarct-Vinfarct, and non-infarct-Vnon-infarct volumes) were completed by domain experts. We used Cohort 1 for brain and infarct segmentation model development (n = 3 training cases with 36 slices [18 anterior and posterior faces], n = 18 testing cases with 218 slices [109 anterior and posterior faces]), as well as infarct morphometrics automation. The infarct quantification pipeline and pre-trained model were packaged as a standalone software and applied to Cohort 2, an internal validation dataset. Finally, software and model trainability were tested as a use-case with Cohort 3, a dataset from a separate institute. Results: Both high segmentation and statistically significant quantification performance (correlation between manual and software) were observed across all datasets. Segmentation performance: Cohort 1 brain accuracy = 0.95/f1-score = 0.90, infarct accuracy = 0.96/f1-score = 0.89; Cohort 2 brain accuracy = 0.97/f1-score = 0.90, infarct accuracy = 0.97/f1-score = 0.80; Cohort 3 brain accuracy = 0.96/f1-score = 0.92, infarct accuracy = 0.95/f1-score = 0.82. Infarct quantification (cohort average): Vbrain (ρ = 0.87, p < 0.001), Vinfarct (0.92, p < 0.001), Vnon-infarct (0.80, p < 0.001), %infarct (0.87, p = 0.001), and infarct:non-infact ratio (ρ = 0.92, p < 0.001). Conclusion: Tectonic Infarct Analysis software offers a robust and adaptable approach for rapid TTC-based stroke assessment.

Biodegradable Redox-Responsive AIEgen-Based-Covalent Organic Framework Nanocarriers for Long-Term Treatment of Myocardial Ischemia/Reperfusion Injury.

Timely restoration of blood supply after myocardial ischemia is imperative for the treatment of acute myocardial infarction but causes additional myocardial ischemia/reperfusion (MI/R) injury, which has not been hitherto effectively targeted by interventions for MI/R injury. Hence, the development of advanced nanomedicine that can reduce apoptosis of cardiomyocytes while protecting against MI/R in vivo is of utmost importance. Herein, a redox-responsive and emissive TPE-ss covalent organic framework (COF) nanocarrier by integrating aggregation-induced emission luminogens and redox-responsive disulfide motifs into the COF skeleton is developed. TPE-ss COF allows for efficient loading and delivery of matrine, a renowned anti-cryptosporidial drug, which significantly reduces MI/R-induced functional deterioration and cardiomyocyte injury when injected through the tail vein into MI/R models at 5 min after 30 min of ischemia. Moreover, TPE-ss COF@Matrine shows a drastic reduction in cardiomyocyte apoptosis and improvements in cardiac function and survival rate. The effect of the TPE-ss COF carrier is further elucidated by enhanced cardiomyocyte viability and triphenyltetrazolium chloride staining in vitro. This work demonstrates the cardioprotective effect of TPE-ss COFs for MI/R injury, which unleashes the immense potential of using COFs as smart drug carriers for the peri-reperfusion treatment of ischemic heart disease with low cost, high stability, and single postoperative intervention.

Dihydromyricetin down-regulating the expression of nod-like receptor protein-3 in the brain tissue of ischemic stroke in rats / 解剖学报

[Abstract] Objective To investigate the role of dihydromyricetin (DHM) in the treatment of ischemic stroke in rats, and to explore the effect of DHM on the expression of inflammasome. Methods The middle cerebral artery occlusion (MCAO) model was induced by endovascular suture method. The therapeutic effect and mechanism of DHM were investigated by Longa score, TTC staining, Nissl staining, immunohistochemical staining and Western bloting. Results After DHM treatment, the motor capacity of MCAO rats was significantly improved, the infarct volume was significantly reduced, the brain structure and neuron morphology were improved, and the expressions of nod-like receptor protein-3 (NLRP3) and interleukin-1(IL-1) decreased significantly. Conclusion DHM can down-regulate the expression of NLRP3 and thus reduces the cerebral infarction volume and improves neurobehavioral performance in MCAO rats.

Stroke core revealed by tissue scattering using spatial frequency domain imaging.

Ischemic stroke leads to a reduction or complete loss of blood supply causing injury to brain tissue, which ultimately leads to behavioral impairment. Optical techniques are widely used to study the structural and functional changes that result as a consequence of ischemic stroke both in the acute and chronic phases of stroke recovery. It is currently a challenge to accurately estimate the spatial extent of the infarct without the use of histological parameters however, and in order to follow recovery mechanisms longitudinally at the mesoscopic scale it is essential to know the spatial extent of the stroke core. In this paper we first establish optical coherence tomography (OCT) as a reliable indicator of the stroke core by analyzing signal attenuation and spatially correlating it with the infarct, determined by staining with triphenyl-tetrazolium chloride (TTC). We then introduce spatial frequency domain imaging (SFDI) as a mesoscopic optical technique that can be used to accurately measure the infarct spatial extent by exploiting changes in optical scattering that occur as a consequence of ischemic stroke. Additionally, we follow the progression of ischemia through the acute and sub-acute phases of stroke recovery using both OCT and SFDI and show a consistently high spatial overlap in estimating infarct location. The use of SFDI in assessing infarct location will allow longitudinal studies targeted at following functional recovery mechanisms on a mesoscopic level without having to sacrifice the mouse acutely.

Elucidation of the enzyme involved in 2,3,5-triphenyl tetrazolium chloride (TTC) staining activity and the relationship between TTC staining activity and fermentation profiles in Saccharomyces cerevisiae.

2,3,5-Triphenyl tetrazolium chloride (TTC) staining is a method to distinguish the mitochondrial activity of cells based on the color: colorless TTC turns red when under reducing conditions. Although the assay reflects the mitochondrial activity of cells, which enzyme(s) in the electron transport system contribute to TTC reduction has been unclear. TTC staining assays using gene disruptants related to the electron transport system in Saccharomyces cerevisiae revealed those disruptants related to electron transport from each electron donor to ubiquinone (red colonies) and disruptants that were related to ubiquinol-cytochrome c oxidoreductase and cytochrome c oxidase (white colonies). In addition, when the enzyme activities of ubiquinol-cytochrome c oxidoreductase and cytochrome c oxidase were measured using TTC as the electron acceptor, only ubiquinol-cytochrome c oxidoreductase showed TTC reduction activity, and the activity was enhanced by potassium cyanide, an inhibitor of cytochrome c oxidase. These results indicated that ubiquinol-cytochrome c oxidoreductase is involved in TTC reduction in S. cerevisiae. The fermentation profiles of BY4741UΔcor1 and BY4741UΔcox4, which exhibited no TTC staining activity, were almost identical to that of the parental strain BY4741U. However, cell growth and ethanol and succinate production of the ura3-mutated strain BY4741, which also exhibited no TTC staining activity, was altered compared to those of BY4741U, indicating that the fermentation profile varies among strains that show no TTC staining activity. The relationship between uracil metabolism and TTC staining activity was also determined based on metabolome analysis.

Brain slice viability determined under normoxic and oxidative stress conditions: involvement of slice quantity in the medium.

Objective: In vitro acute adult brain slice methods are instruments in developing our knowledge of the nervous system. Optimization of this method for obtaining high-quality brain slices is extremely important in terms of consistency and reliability of the experimental results. Although some important topics such as slice thickness, temperature, and composition of the physiological medium have been studied for optimization, involvement of slice quantity in medium on tissue viability has not been investigated yet.Methods: Different number of slices (1, 3, or 6 slices) were incubated under normoxic or some prooxidant stress conditions induced by oxygen-glucose deprivation (OGD), H2O2, FeSO4+ ascorbic acid, or menadione to evaluate the effect of slice density on tissue viability.Results:Slice quantity in the normoxic incubation medium caused a significant increase in 2,3,5-triphenyltetrazolium chloride (TTC) staining intensity of the slices. Similarly, increase in the slice quantity in the medium also protected the slices against either OGD, H2O2, FeSO4, or menadione-induced decrease in TTC staining. In addition to TTC staining, lactate dehydrogenase leakage or malondialdehyde and reactive oxygen species production under normoxic or ischemia-like conditions were also attenuated by increasing slice quantity in the medium.Conclusion: These results show that when using brain slices method for investigating the structural and functional features of brain at the molecular and cellular levels, both slice quantity in the medium and incubation volume should be considered first. Increasing slice quantity or decreasing incubation volume probably causes an increase in the concentration of endogenous substance(s) involved in neuroprotection.

Establishment of mouse myocardial infarction model and early electrocardio- gram assessment / 药学实践杂志

Objective To establish and optimize a mouse myocardial infarction (MI) model, and to use twice limb lead ECGs immediately after coronary ligation and 4 h after surgery to evaluate the occurrence of myocardial infarction. Methods Twenty-nine male C57BL/6J mice were anesthetized with isoflurane. then a myocardial infarction model was established by ligating the left anterior descending (LAD) coronary artery through the third/fourth intercostal space of left anterior chest. Immediate and 4 h postoperative limb lead ECGs were performed. Twenty-four hours after surgery, the chest was opened and the occurrence of myocardial infarction was evaluated. The heart samples were taken for TTC staining to determine the infarct area and calculate the infarct area. Results During the mice underwent coronary artery ligation the intraoperative mortality was 6.8% (2/29), and the early postoperative (<4 h) mortality was 10.3% (3/29). The 24 h survival rate was 82.8% (24/29). 24 hours after TTC staining confirmed the occurrence of infarction, the myocardial infarction model was established. The success rate of the model was 79.3% (23/29), and the average infarct size (infarcted myocardial weight / whole ventricular weight) was (28 ± 6)%; The mice successfully established by the model showed obvious ST-T changes in the ECG at 4 hours after surgery, suggesting that a myocardial infarction has occurred. Conclusions The mouse myocardial infarction model was successfully established. The combined use of ECG immediately after surgery and 4 h after surgery could be used as a rapid and non-invasive evaluation method for mouse myocardial infarction.

Neural like cells and acetyl-salicylic acid alter rat brain structure and function following transient middle cerebral artery occlusion.

Introduction Transient cerebral ischemia is a pandemic neurological disorder and the main aim of medical intervention is to reduce complications. Human umbilical cord mesenchymal cells (hUCMs) are capable of differentiating into neural-like cells (NLC) in vitro, therefore we investigated the neuroprotective potential of these cells in comparison to aspirin and in combination (NLC-Aspirin) on spatial memory and neural morphologic changes in male rats submitted to transient cerebral ischemia. Methods Ten days after the intervention, the improvement in learning and memory were assessed in the animals by Morris Water Maze. Thence, the animals were examined for the presence of PKH26 labeled cells in the ischemic area of the brain, the infarct volume and neural changes in the brain tissue. Results Significant spatial memory deficits in the ischemic animals were detected compared with the control animals. The learning and memory were significantly improved (p ≤ 0.05) in the aspirin and NLC groups compared with the ischemic animals. Co-treatment of aspirin and NLCs did not improve the outcome. Moreover, infarction volume and neural changes were significantly altered when aspirin or NLCs were administered. Conclusions Our data suggest the significant neuroprotective potential of aspirin and neural-like cells derived from hUCM cells in the treatment of brain ischemic stroke. Further studies are required to evaluate possible underlying mechanisms, and to evaluate the possible interactions between aspirin and stem cells in a joint treatment aimed at the recovery of cognitive impairments.

Use of 2,3,5-triphenyltetrazolium chloride-stained brain tissues for immunofluorescence analyses after focal cerebral ischemia in rats.

The middle cerebral artery occlusion (MCAO) model in rodents has been widely used as model for studying brain ischemic stroke. TTC (2,3,5-triphenyltetrazolium chloride) staining in fresh tissues is used to evaluate the size of the infarct in MCAO model, and TTC-stained brain tissues are considered to be possible to bring a damage to the anatomical structure of neuronal cells and unsuitable for immunofluorescence analyses of cytology, and discarded after evaluation of infarct volume. Another group of models with in vivo fixation was required to the pathological or histological analyses of the infarct brains, which lead to double the numbers of animals in researches. However, some evidences indicate that if we properly optimized staining protocol, TTC-stained brain tissues might be suitable for cytological analyses. In this work, we have optimized the immunofluorescent staining methods of TTC-stained brain slices, and found that TTC-stained brain tissues are suitable for quantitative and qualitative analyses of microglia, astrocytes and neuroblasts, the morphology of theses cell were nearly identical to the in-vivo fixed models. Our optimized-protocol provide two advantages over traditional methods one of them is providing the precise the infarct region, which reduces the differences within groups, the other one is decreasing the total number of animals in research dramatically.

Detection of hypothalamic infarction in intraluminal middle cerebral artery occlusion model rat by DWI scanning / 上海交通大学学报（医学版）

Objective: To detect and evaluate the hypothalamic infarction in middle cerebral artery occlusion (MCAO) model rat. Methods: For 15 Sprague-Dawley rats weighed 200-250 g, aged 6-8 months, their right middle cerebral artery was occluded for 90 min by a silicon-coated 4-0 nylon filament and reperfused. Sprague-Dawley rats underwent diffusion weighted MR imaging (DWI) scanning (at 1 h and 24 h after reperfusion) and 2, 3, 5-triphenyl tetrazolium chloride (TTC) staining (at 24 h after reperfusion) to determine the hypothalamic and cerebral infarct volume. The relationship between hypothalamic infarct volume and cerebral infarction volume was analyzed by DWI scanning. The results of TTC staining were compared with those of 24 h DWI scanning. Results: Fifteen Sprague-Dawley rats successfully received intraluminal MCAO/reperfusion procedures. The incidences of hypothalamic infarction on brain DWI scanning and TTC staining were 100% and 40% at 24 h after reperfusion, respectively. Therefore, DWI scanning was more sensitive than TTC staining to detect hypothalamic injury (P=0.001). The hypothalamic infarct volume on DWI scanning was (8.59±2.89) mm3 and (11.65±3.19) mm3 at 1 h and 24 h after reperfusion, respectively. On DWI scanning, hypothalamic and cerebral infarct volume at 24 h after reperfusion were correlated with each other significantly (r=0.573, P=0.025), so were the increases of hypothalamic and cerebral infarct volume (r=0.554, P=0.032) from 1 h to 24 h. Conclusion: DWI scanning was more sensitive than TTC staining to detect hypothalamic injury in intraluminal transient MCAO model. Hypothalamic and cerebral infarct volume were correlated with each other.

A Simple Procedure for the Evaluation of Bone Vitality by Staining with a Tetrazolium Salt.

Presently, no intra-operative method for a direct assessment of bone vitality exists. Therefore, we set out to test the applicability of tetrazolium-based staining on bone samples. The explanted femoral heads of 37 patients were used to obtain either cancellous bone fragments or bone slices. Samples were stained with 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (thiazolyl blue, MTT) at different times (one to twelve hours) after explantation. Staining was quantified either spectrophotometrically after extraction of the dyes or by densitometric image analysis. TTC-staining of cancellous bone fragments and bone slices, respectively, indicated the detectability of vital cells in both types of samples in a window of up to six hours after explantation. Staining intensity at later time-points was indistinguishable from the staining of untreated samples or sodium azide treated samples, which represent dead cells. In contrast, MTT-staining of bone slices revealed intense unspecific staining, which obscured the evaluation of the vitality of the samples. The lack of a detectable increase of colour intensity in TTC-stained bone samples, which were treated more than six hours after explantation, corresponds to reduced fracture healing. The described simple procedure could provide a basis for an intraoperative decision by the orthopaedic surgeon.

Evaluation of a vital staining protocol with 2,3,5-triphenyltetrazolium chloride for cancellous bone in a sheep model.

Decision making on the optimal surgical treatment of fractures often is hampered by the lack of a method for direct assessment of bone vitality. In various contexts, for example to determine the extents of cerebral insults or of myocardial infarctions in experimental studies, tetrazolium based staining procedures of vital cells are widely used. Here, we set out to test the applicability of tetrazolium based staining on bone samples. 8 brains and 26 femoral heads from sheep were used to prepare tissue slices, which were stained with 2,3,5-triphenyltetrazolium chloride (TTC) at various times (1 to 12h) after explantation. Staining of tissue slices was quantified by densitometric image analysis. Spectrophotometry was used for quantification in cultured cells. TTC-staining of tissue slices indicated detectability of vital cells in slices from both tissues up to 4h after explantation. Staining intensity at later time-points was indistinguishable from the staining of untreated samples or sodium azide treated (necrotic cells) controls. We provide experimental evidence that the choice of the optimal surgical approach for the treatment of fractures involving cancellous bone could be aided by a simple staining procedure for vital bone. However, the described procedure depends on the availability of bone specimens (slices). Therefore, search for an improved stain directly applicable to the bone surface is needed.

Protection of Batroxobin Injection on super acute stage of thromboembolic cerebral stroke in rats / 药物评价研究

Objective To observe the effects of Batroxobin Injection on thromboembolic cerebral stroke by magnetic resonance imaging (MRI) and TTC staining.Methods Rat model ofthromboembolic stroke was prepared after the left middle cerebral artery was occluded by autologous blood clots,and 32 rats with successful operation were divided into four groups according to the degree of neurological deficit:model group,Batroxobin Injection low and high dose (0.3,1.0 BU/kg) group,and rt-PA (9 mg/kg) group,with eight rats in each group,and other eight rats in Sham group.Rats were administered 1 h after modeling by tail iv method.At 6 h after administration,neurological deficit score and MRIincluding SE-T2WI and DWI sequence scanning were measured.At 24 h after administration,the brain was cut for TTC staining to measure the infarct area,and blood FIB was measured.Results Compared with model group,Batroxobin Injection 0.3 BU/kg treatment for 24 h (P ＜ 0.05),1 BU/kg treatment for 6 and 24 h (P ＜ 0.05,0.01) could significntly improve the neurological function scores of rats.MRIresults showed that Batroxobin Injection at dose of 0.3 and 1 BU/kg significantly reduced the lesion range (P ＜ 0.05 and 0.01).Results of TTC stain showed that Batroxobin Injection at dose of 0.3 and 1 BU/kg significantly reduced the infarct size (P ＜ 0.05).Batroxobin Injection at doses of 0.3 and 1 BU/kg can significantly lower plasma FIB concentration (P ＜ 0.05,0.01,0.001) 6 and 24 h after administration.Conclusion Batroxobin Injection can improve the damaged neural function,reduce scope of lesions,decrease plasma fibrinogen,with protective effects for cerebral ischemia in rats.

Protection of Batroxobin Injection on super acute stage of thromboembolic cerebral stroke in rats / 药物评价研究

Objective To observe the effects of Batroxobin Injection on thromboembolic cerebral stroke by magnetic resonance imaging (MRI) and TTC staining.Methods Rat model ofthromboembolic stroke was prepared after the left middle cerebral artery was occluded by autologous blood clots,and 32 rats with successful operation were divided into four groups according to the degree of neurological deficit:model group,Batroxobin Injection low and high dose (0.3,1.0 BU/kg) group,and rt-PA (9 mg/kg) group,with eight rats in each group,and other eight rats in Sham group.Rats were administered 1 h after modeling by tail iv method.At 6 h after administration,neurological deficit score and MRIincluding SE-T2WI and DWI sequence scanning were measured.At 24 h after administration,the brain was cut for TTC staining to measure the infarct area,and blood FIB was measured.Results Compared with model group,Batroxobin Injection 0.3 BU/kg treatment for 24 h (P ＜ 0.05),1 BU/kg treatment for 6 and 24 h (P ＜ 0.05,0.01) could significntly improve the neurological function scores of rats.MRIresults showed that Batroxobin Injection at dose of 0.3 and 1 BU/kg significantly reduced the lesion range (P ＜ 0.05 and 0.01).Results of TTC stain showed that Batroxobin Injection at dose of 0.3 and 1 BU/kg significantly reduced the infarct size (P ＜ 0.05).Batroxobin Injection at doses of 0.3 and 1 BU/kg can significantly lower plasma FIB concentration (P ＜ 0.05,0.01,0.001) 6 and 24 h after administration.Conclusion Batroxobin Injection can improve the damaged neural function,reduce scope of lesions,decrease plasma fibrinogen,with protective effects for cerebral ischemia in rats.

Hyperglycemia aggravated cerebral ischemia/reperfusion injury by inhibiting mitophagy / 中国药理学通报

Aim Toinvestigatetheroleofhyperglyce-mia in cerebral ischemia/reperfusion(I/R)injury with a middle cerebral artery occlusion(MCAO)rat model anditsmechanism.Methods EightyhealthymaleSD rats were randomly assigned into sham group, I/R group (normoglycemia),hyperglycemic I/R groupⅠ(HG1 )and hyperglycemic I/R groupⅡ(HG2 ).The cerebral I/R model was established by occluding the middle cerebral artery(MCA)in rats.Hyperglycemia was induced by intraperitoneal injection of 50% glu-cose solution.Neurological deficit was determined by Ludmila Belayev test;infarct size and brain edema were measured by TTC staining;mitophagy was ob-served by double immunofluorescent staining and elec-tron microscope.The expressions of autophagy-related proteins(LC3 and Beclin-1 )and apoptosis-related pro-teins(Cyt-C,AIF,caspase-9 and caspase-3 )were ex-aminedbyWesternblotfurtherly.Results Bloodglu-cose level was controlled at 4 mmol·L-1 (normoglyce-mia),10 mmol · L-1 (HG1 ) and 20 mmol · L-1 (HG2)respectively.There were no significant differ-ences between model group and HG1 group in neuro-logical deficit scores,infarct volume and edema size(P>0. 05 ).However,these indications in HG2 group were significantly increased compared with model group (P<0. 05 ).After 3 days of reperfusion,the level of mitophagy was significantly reduced accompanied with increased mitochondria damages in HG 2 group (P <0. 05 ),and the expressions of mitochondrial related ap-optotic proteins(Cyt-C,AIF,caspase-9 and caspase-3 ) were significantly increased accordingly compared to modelgroup.Conclusions Mildhyperglycemiacan not intensify the cerebal ischemic injury.In contrast, severe hyperglycemia significantly aggravates the brain ischemic injury by inhibiting the removal of injured mi-tochondria in a manner of mitophagy,thus amplifying the mitochondrial mediated cascade damage responses.

Stromal Cell-Derived Factor-1α Plays a Crucial Role Based on Neuroprotective Role in Neonatal Brain Injury in Rats.

Owing to progress in perinatal medicine, the survival of preterm newborns has markedly increased. However, the incidence of cerebral palsy has risen in association with increased preterm birth. Cerebral palsy is largely caused by cerebral hypoxic ischemia (HI), for which there are no effective medical treatments. We evaluated the effects of stromal cell-derived factor-1α (SDF-1α) on neonatal brain damage in rats. Left common carotid (LCC) arteries of seven-day-old Wistar rat pups were ligated, and animals were exposed to hypoxic gas to cause cerebral HI. Behavioral tests revealed that the memory and spatial perception abilities were disturbed in HI animals, and that SDF-1α treatment improved these cognitive functions. Motor coordination was also impaired after HI but was unimproved by SDF-1α treatment. SDF-1α reduced intracranial inflammation and induced cerebral remyelination, as indicated by the immunohistochemistry results. These data suggest that SDF-1α specifically influences spatial perception abilities in neonatal HI encephalopathy.

Coadministration of the Human Umbilical Cord Matrix-Derived Mesenchymal Cells and Aspirin Alters Postischemic Brain Injury in Rats.

BACKGROUND: Ischemic stroke is an acute brain insult that induces dramatic changes in the neurons. Treatment of brain stroke is one of the main therapeutic targets of neuroprotective therapies. The aim of this study was to evaluate the protective potential of implanted human umbilical cord mesenchymal stem (hUCMs) cells with/without aspirin (ASA) against focal cerebral ischemia. METHODS: We assessed the migration and distribution of PKH26-labeled cells after transplantation. After day 10 of transient occlusion, we evaluated the effect of ASA and hUCMs on the recovery of learning and memory in rats by Morris water maze. Afterward, animals were sacrificed, and the infarct area in the brain was evaluated using 2, 3, 5-triphenyltetrazolium chloride staining and also by hematoxylin and eosin. RESULTS: The recovery of learning and memory in ischemic animals that received ASA and hUCM cells improved significantly compared with the untreated ischemic animals. Coadministration of ASA and hUCM cells did not improve the outcome at a comparable rate with ASA and hUCM cells alone. PKH26-labeled cells were detectable in the ischemic area of the brain tissue sections. 2,3,5-Triphenyltetrazolium chloride staining and histologic examinations showed that treatment with ASA and hUCM cells could significantly alter the ischemic area. CONCLUSIONS: The results of the present study suggest that ASA and hUCM cells can withstand degenerative changes induced by artificial stroke in the rat. Also the learning and memory disturbance in the ASA and cell-treated animals is less pronounced than ischemic animals. Coadministration of ASA and hUCM cells did not raise the outcome higher than administration of ASA and hUCM cells alone.

Pharmacological modulation of I(1)-imidazoline and α2-adrenoceptors in sub acute brain ischemia induced vascular dementia.

Sub-acute brain ischemia is a risk factor for the development of vascular dementia (VaD). Sub-acute brain ischemia induced VaD, participates in a negative role in impaired cognition. Imidazoline receptors are widely expressed in the central nervous system. But the role of I1-imidazoline and α2-adrenoceptors in VaD are still unknown. The present study has been designed to investigate the role of selective I1-imidazoline receptor modulator; moxonidine as well as α2-adrenoceptor modulator; clonidine in sub-acute brain ischemia induced VaD in mice (n=8). Permanent bilateral common carotid arteries ligation (2VO) technique was used to induce sub-acute brain ischemia in mice. Assessment of spatial learning and memory was done by using Morris water maze. Brain damage was assessed as percent infarct, using TTC staining of brain coronal sections. Oxidative stress was assessed by estimating brain malondialdehyde (MDA), catalase (CAT), glutathione (GSH) and superoxide dismutase (SOD). Cholinergic status was assessed by brain acetylcholinesterase (AChE) activity. 2VO animals have shown significant reduction in learning and memory as well as brain CAT, GSH and SOD, with significant increase in brain infarct size, MDA and AChE activity. Whereas, administration of moxonidine and clonidine significantly attenuated 2VO induced learning and memory deficits, brain damage, brain oxidative stress and higher AChE activity. It may be concluded that 2VO induced sub-acute brain ischemia has elicited dementia, which was attenuated by moxonidine and clonidine. Thus, modulators of I1-imidazoline receptors may be explored further for their benefits in sub-acute brain ischemia induced vascular dementia.