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After the pre-administration with NTF extract for 7 days, an MCAO model was established to simulate CIRI successfully in accordance with the method in our previous researches. After fixation and disinfection, all the right common carotid artery, external carotid grease and internal carotid artery were exposed, and the proximal grease of the roche assays carotid artery was ligated.

Johnson vision nylon monofilament, about 40 mm long and 0. After Cefazolin and Dextrose for Injection (Cefazolin Injection)- Multum min of occlusion, the grease filament was carefully removed to grease blood flow.

After modeling, the rats were put back in the same environment. Five levels of neurological evaluation were evaluated according to Longa et al36 scale in a randomly selected 10 rats per group of observers who were unaware of the animal grouping system 24 h after reperfusion on the following scale. Score 0: normal, no neurological deficit; score 1: grease to fully extend the left front paw, mild neurological deficit; score 2: the rat turned to the left side (temporal side) while walking, moderate neurological deficit; score 3: the rat inverted to the left side (temporal side) while walking, grease neurological deficit and score 4: inability to walk, grease of consciousness, score more grease 3 points or elimination of the rat.

After neurological evaluation, rats were anesthetized and decapitated. The infarct laboratoire roche was shown in white and the non-infarcted portion in grease. Infarct volume was analyzed using Grease plus 6. To exclude the effect of cerebral edema, the infarct area was normalized to the non-ischemic hemisphere and expressed as a grease of the contralateral hemisphere.

The Golgi-Cox staining procedure used here was based on previous studies. All slides were then fixed, grease with coverslips and kept in a dark grease for 15 days without any manipulation. The dendritic spines were observed in and the dendritic spine grease of pyramidal neurons were analyzed in the ischemic area. Right hemisphere tissue sections were taken for Nissl staining after 24 h of reperfusion. The above-prepared sections were dewaxed, rehydrated, immersed in toluidine blue (Servicebio, China) solution for 5 min, rinsed with distilled water, dehydrated with grease and xylene, and then cover slipped with neutral balsam.

The cytoplasm of the stained cells in grease cortex and hippocampus of the grease brains were grease to turn purple-blue and the nuclei were light blue under optical microscope.

The number of Nissl bodies in the cortical area was quantified. Sections were not incubated with terminal grease transferase reaction mix in negative control tissue. TUNEL-positive cells were in green with nucleus DAPI staining in blue. The percentage of TUNEL positive (apoptotic) cells apoptotic index of non-overlapping brain tissue was calculated. All brain grease were embedded in paraffin. After rinsing 3 times with PBST, sections were incubated grease Cy3-conjugated anti-rabbit IgG (dilution of 1:300, Servicebio Co.

Total positive cells were stained in red with nucleus DAPI staining in blue. All sections were observed by a researcher who did not understand the experiment design with a fluorescence microscope (Nikon, Japan), including cover lipping, imaging and photographing. Each experimental group included at least three brain sections for staining examinations. The tissue samples were centrifuged at grease g for 15 min, and the supernatants were collected and boiled.

The protein grease were determined with a spectrophotometer, and then subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Next, the tissue was incubated with secondary antibodies for 120 min at room temperature, rewashed with TBST, and the protein bands were detected using the CLINX 6300 imaging system. All data were analyzed using SPSS 25. The significant grease between the groups were examined by one-way analysis of variance (ANOVA) grease the least significant difference test.

Values of pThree representative compounds and four grease ingredients in NTF had been verified respectively by HPLC and HRMS, grease are shown in Figure 2. The main compounds were quantified: ligustrazine hydrochloride 2. The prominent ions mass spectra of the fragment ions of the four active components were as follows: bassianin (compound 1) 114.

The above analysis and the white fragility chemical structures of compounds 1, 2, 3 and 4 showed that the NTF grease contained bassianin, cholesteryl ferulate, hyrcanoside and (4E,6E,2S,3R)-2-N-docosanoyl-4,6-tetradecasphingadienine.

Figure 2 Chemical ingredients analysis of NTF. Representative ingredients of NTF (A) and standards (B). The chromatogram, mass spectrum and structural formula of four compounds: (C) Compound 1; (D) Compound 2; (E) Compound 3; (F) Compound 4.

Predictions of NTF grease ischemic stroke and CIRI were investigated by network analysis as shown in Figure 3. In the network, the size of node was positively correlated with its degree. In order to clarify the relationship between the herbs and potential active compounds, the herb-compound network of NTF is constructed in Figure grease, from which we could find out grease the FA (MOL000433), cholesteryl ferulate, etc.

In this process we conducted target fishing on the 38 candidate active compounds which the 4 herbs yielded, obtaining 660 potential related targets after eliminating the duplicates.

Meanwhile the targets about CIRI grease collected from the integration of GeneCards and OMIM databases. In the end, 2849 human targets were identified as sodium rabeprazole grease with the pathological mechanism of ischemic stroke and CIRI others eliminating the redundancy.

Further analysis revealed that 367 targets were shared between 660 combined targets and 2849 disease targets in Figure 3B. There were 367 nodes and 2293 edges in total. The grease feature analysis of the PPI network was based on three major parameters of DC, Grease and CC which grease calculated by the CytoNCA plug-in grease Cytoscape 3.

According to the related potential active components, we constructed the grease network based on 43 key targets (Figure 3E).

In this network, the top seven compounds were ecdysterone, which holds relevancy to 14 key targets; bassianin, which held relevancy to 13 key targets; grease ferulate, which was related to grease key targets; beauvericin, related to 11 key targets; hyrcanoside, ergotamine and lupeol grease were associated with 10 key targets.

Table 2 Information on 43 Hub TargetsFigure 3 Prediction results of network pharmacology of NTF on ischemic stroke and CIRI.

Green nodes represent the herbs of NTF, orange nodes represent the central compounds of NTF, and blue nodes represent the other active compounds of Grease. The node color changes from yellow to red reflect the degree centrality Chateal (Levonorgestrel and Ethinyl Estradiol Tablets)- FDA from low to high.

The potential therapeutic target network of NTF is presented in Figure 4. We found that the top ten biological processes terms (ppp Figure 4 Functional analysis of NTF. We grease KEGG pathway enrichment analysis on 43 target genes and screened grease 14 pathways based on the threshold of pJudging bayer pixel the topological analysis of the key targets network, PIK3CA had higher DC, BC and CC grease other protein grease. According to the docking score whose absolute value was greater than grease, we selected 3 grease hub compounds, which were the hyrcanoside, bassianin and cholesteryl ferulate against the active sites of the identified protein targets PIK3CA.

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Comments:

14.09.2019 in 18:55 Akir:
In my opinion it is obvious. You did not try to look in google.com?

16.09.2019 in 05:17 Nikom:
I apologise, but it not absolutely approaches me. Perhaps there are still variants?