To investigate the effects and mechanism of qi-tonifying and stasis-eliminating (QTSE) therapy on the expression of vascular endothelial growth factor (VEGF) and its receptors Flt-1 and Flk-1 in the ...brains of intracerebral hemorrhagic (model) rats.
One hundred and eighty Sprague-Dawley rats were randomly divided into six groups: the normal group (n=5), the sham-operative (SO) group (n=35), the model group (n=35), the QTSE group (n=35), the QT group (n=35) and the SE group (n=35). All the rats except those in the normal group and SO group were established into an intracerebral hemorrhage(ICH) model by intracerebral injection of collagenase type VII and the latter three were orally administered with Buyang Huanwu Decoction (a classical recipe for QTSE) or with some of its components for qi-tonification and for stasis-elimination, respectively. To the other three groups, normal saline solutions were given instead. Behavioral tests were carried out in the animals randomly chosen from each group on days 1, 2, 4, 7, 14, 21 and 28 after modeling. The expressions of VEGF, Flk-1 and Flt-1 were determined by immunohistochemistry and the number of vascular segments with positive expression in the injured brain area of the rats was calculated.
From day 7 onwards, the asymmetric forelimb use rate in the QTSE group recovered more significantly than that in the other model groups. In the model group, the expressions of VEGF, Flk-1 and Flt-1 appeared on day 1 and reached a peak on day 21, then weakened gradually. In the QTSE group, as compared with the other model groups, a higher level of VEGF expression was shown from day 7 (P<0.01) and a higher level of Flt-1 expression was shown from the 7th day to the 21st day (P<0.01).
QTSE therapy can up-regulate the expressions of VEGF and its receptors (Flk-1 and Flt-1) and improve the recovery of kinetic function in the ICH rats, which may be correlated with its action in modulating vascular regeneration to promote the reconstruction of microvascular networks in the damaged areas.
The purpose of this study was to investigate the effect of inhibition of calpain on retinal ganglion cell-5 (RGC-5) necroptosis following oxygen glucose deprivation (OGD). RGC-5 cells were cultured ...in Dulbecco’s-modified essential medium and necroptosis was induced by 8-h OGD. PI staining and flow cytometry were performed to detect RGC-5 necrosis. The calpain expression was detected by Western blotting and immunofluorescence staining. The calpain activity was tested by activity detection kit. Flow cytometry was used to detect the effect of calpain on RGC-5 necroptosis following OGD with or without N-acetyl-leucyl-leucyl-norleucinal (ALLN) pre-treatment. Western blot was used to detect the protein level of truncated apoptosis inducing factor (tAIF) in RGC-5 cells following OGD. The results showed that there was an up-regulation of the calpain expression and activity following OGD. Upon adding ALLN, the calpain activity was inhibited and tAIF was reduced following OGD along with the decreased number of RGC-5 necroptosis. In conclusion, calpain was involved in OGD-induced RGC-5 necroptosis with the increased expression of its downstream molecule tAIF.
To explore the effect of precursor of brain-derived neurotrophic factor (proBDNF) on cultured hippocampal neuron and its intracellular mechanism.
The hippocampal neurons were dissociated from E18 ...rats and cultured in neurobasal medium, and then the cells were treated with proBDNF, preBDNF(propeptide of proBDNF) and proBDNF antiserum,respectively. Thirty minutes, 1 hour or 48 hours later, the cells were stained with Nissl solution, and the immunocytochemistry methods of ELK-p (Ets domain protein), ErK2(extracellular signal regulated kinase) and c-fos were performed.
The expressions of ELK-p, ErK and c-fos were significantly upregulated in the cultured hippocampal neurons after they were treated with proBDNF protein,and the immuno-positive staining was also obvious in some nuclei. While the endogenous proBDNF was neutralized by proBDNF antiserum treatment, the expressions of ELK-p, ErK and c-fos were downregulated and many cells showed swelling and vasoculation. The immunoreactivity in preBDNF treated cells was
To study the effects of selective head mild hypothermia on endogenous neuroprotection in brain following global cerebral ischemia/reperfusion.
Fifteen dogs were randomly divided into three groups: ...nonischemic control group (Group A, n = 4), undergoing thoracotomy without cerebral ischemia and general care for 8 hours thereafter; cerebral ischemia/reperfusion group (Group B, n = 5) undergoing thoracotomy, clipping of the ascending aorta for 18 min, cardiac resuscitation, maintenance of respiration and circulation for 8 h; and mild hypothermia group (Group C, n = 6), received selective head mild hypothermia, i.e. lowering the tympanic temperature to (34 +/- 0.5) degrees C for 8 hours after cerebral ischemia. The neurological function was assessed by Glasgow coma scale and Pittsburgh brain stem score. At the end of experiment, the dog brains were taken out to obtain the right parietal cerebral cortex. Immunohistochemistry was used to detect the parvalbumin (PV) and HSP(70). Xanthine oxidase method was used to de
Stem cells transplant of retina Chen, Dan; Huang, Ju-Fang; Luo, Xue-Gang
Zhong nan da xue xue bao. Journal of Central South University. Yi xue ban
29, Številka:
3
Journal Article
In this study, the 3'untranslated region (3'UTR) of MHC class I chain-related gene B (MICB) were investigated in 104 healthy, unrelated Han individuals recruited from northern China, using ...PCR-sequencing method. Seven polymorphic sites were detected including a 2-bp deletion/insertion, a 6-bp deletion/insertion and 5 SNPs. Seven different 3'UTR alleles were identified with frequencies ranging from 0.0048 to 0.7981. MICB(∗)005:02, the most frequent allele in this population, exhibited significant linkage disequilibrium (LD) with UTR1; MICB(∗)004:01, which was also relatively common in this population, showed strong LD with both UTR2 and UTR3 alleles. Analysis for targets of miRNAs revealed that miRNA hsa-miR-4768-5p, whose seed region binds to positions 82-88 of MICB 3'UTR, encompasses the +11800 A/G polymorphism. Ewens-Watterson homozygosity statistics at MICB coding and 3'UTR regions were consistent with neutral expectations. Phylogenetic analysis demonstrated the existence of two main MICB lineages. Our results provided for the first time the data of genetic variation in the 3'UTR of MICB gene in human populations. The findings are valuable for future studies of the mechanisms underlying MICB post-transcriptional regulation, the potential role of regulatory region of MICB gene in disease susceptibility in related ethnic groups, and will inform studies of evolution of the MHC gene complex.
BackgroundGlaucoma mainly induced by increased intraocular pressure (IOP), it was believed that the pressure that wall of eyeball withstands were determined by material properties of the tissue and ...stereoscopic geometry of the eyeball. In order to study the pressure changes in different parts of interior eyeball wall, it is necessary to develop a novel eye ball FEM with more accurate geometry and material properties. Use this model to study the stress changes in different parts of eyeball, especially the lamina cribrosa (LC) under normal physiological and pathological IOP, and provide a mathematical model for biomechanical studies of selected retinal ganglion cells (RGCs) death.Methods(1) Sclera was cut into 3.8-mm wide, 14.5-mm long strips, and cornea was cut into 9.5-mm-wide and 10-mm-long strips; (2) 858 Mini BionixII biomechanical loading instrument was used to stretch sclera and cornea. The stretching rate for sclera was 0.3 mm/s, 3 mm/s, 30 mm/s, 300 mm/s; and for cornea were 0.3 mm/s and 30 mm/s. The deformation-stress curve was recorded; (3) Naso-temporal and longitudinal distance of LC were measured; (4) Micro-CT was used to accurately scan fresh bovine eyes and obtain the geometrical image and data to establish bovine eye model. 3-D reconstruction was performed using these images and data to work out the geometric shape of bovine eye; (5) IOP levels for eyeball FEM was set and the inner wall of eyeball was used taken as load-bearing part. Simulated eyeball FE modeling was run under the IOP level of 10 mmHg, 30 mmHg, 60 mmHg and 100 mmHg, and the force condition of different parts of eyeball was recorded under different IOP levels.Results(1) We obtained the material parameters more in line with physiological conditions and established a more realistic eyeball model using reversed engineering of parameters optimization method to calculate the complex nonlinear super-elastic and viscoelastic parameters more accurately; (2) We observed the following phenomenon by simulating increased pressure using FEM: as simulative IOP increased, the stress concentration scope on the posterior half of sclera became narrower; in the meantime, the stress-concentration scope on the anterior half of scleral gradually expanded, and the stress on the central part of LC is highest.ConclusionAs simulative IOP increased, stress-concentration scope on the posterior half of sclera gradually narrowed; in the meantime, the stress-concentration scope on the anterior half of sclera gradually expanded, and the stress on the LC is mainly concentrated in the central part, suggesting that IOP is mainly concentrated in the anterior part of the eyeball as it increases. This might provide a biomechanical evidence to explain why RGCs in peripheral part die earlier than RGCs in central part under HIOP.
Chronic lead (Pb) poisoning remains an environmental risk especially for the pediatric population, and it may affect brain development. Immature neurons expressing doublecortin (DCX+) exist around ...cortical layer II in various mammals, including adult guinea pigs and humans. Using young adult guinea pigs as an experimental model, the present study explored if chronic Pb exposure affects cortical DCX + immature neurons and those around the subventricular and subgranular zones (SVZ, SGZ).
Two month-old guinea pigs were treated with 0.2% lead acetate in drinking water for 2, 4 and 6 months. Blood Pb levels in these animals reached 10.27 ± 0.62, 16.25 ± 0.78 and 19.03 ± 0.86 μg/dL at the above time points, respectively, relative to ~3 μg/dL in vehicle controls. The density of DCX + neurons was significantly reduced around cortical layer II, SVZ and SGZ in Pb-treated animals surviving 4 and 6 months relative to controls. Bromodeoxyuridine (BrdU) pulse-chasing studies failed to find cellular colocalization of this DNA synthesis indicator in DCX + cells around layer II in Pb-treated and control animals. These cortical immature neurons were not found to coexist with active caspase-3 or Fluoro-Jade C labeling.
Chronic Pb exposure can lead to significant reduction in the number of the immature neurons around cortical layer II and in the conventional neurogenic sites in young adult guinea pigs. No direct evidence could be identified to link the reduced cortical DCX expression with alteration in local neurogenesis or neuronal death.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Previous studies have demonstrated that doublecortin-positive immature neurons exist pre- dominantly in the superficial layer of the cerebral cortex of adult mammals such as guinea pigs, and these ...neurons exhibit very weak properties of self-proliferation during adulthood under physiological conditions. To verify whether environmental enrichment has an impact on the proliferation and maturation of these immature neurons in the prefrontal cortex of adult guinea pigs, healthy adult guinea pigs were subjected to short-term environmental enrichment. Animals were allowed to play with various cognitive and physical stimulating objects over a period of 2 weeks, twice per day, for 60 minutes each. Immunofluorescence staining results indicated that the number of doublecortin-positive cells in layer II of the prefrontal cortex was significantly increased after short-term environmental enrichment exposure. In addition, these doublecortin-positive cells co-expressed 5-bromo-2-deoxyuridine （a marker of cell prolifera- tion）, c-Fos （a marker of cell viability） and NeuN （a marker of mature neurons）. Experimental findings showed that short-term environmental enrichment can induce proliferation, activation and maturation of doublecortin-positive cells in layer II of the prefrontal cortex of adult guinea pigs.
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