Application of flow cytometry for evaluating clinical prognosis and histopathological grade of human glioma

<p><b>Objectives:</b> Flow cytometry was applied to predict the biological parameters of tumor behavior based on the DNA content distribution of tumors. We used flow cytometry to determine the number of cell cycles for the characterization of intracranial gliomas and its possible prognostic role.</p> <p><b>Methods:</b> Flow cytometric analysis of the DNA content was performed for 37 fresh operative glioma specimens. The expression of Ki-67 in glioma specimens was detected using immunohistochemistry staining. The check points of G2/M-phase fractions, cyclin B, and pCdk1 (Y15) were analyzed using Western immunoblotting.</p> <p><b>Results:</b> Compared to low-grade (grade I/II) gliomas, significant differences in the Ki-67, cyclin B, G2/M-phase, and S+G2/M-phase expressions were found in high-grade (grade III/IV) gliomas. Furthermore, receiver operating characteristic (ROC) analysis indicated optimal cutoff points for the G2/M-phase and S+G2/M-phase fractions of 13.47 and 17.26%, respectively, which can be used to differentiate cases with low- and high-grade gliomas. Additionally, both G2/M-phase and S+G2/M-phase fractions had significant association with the expression of Ki-67 in the gliomas. The gliomas were classified by the DNA content. We found that patients with high-grade glioma had worse survival rate than patients with low-grade glioma. Meanwhile, ROC curve analysis gave cutoffs for G2/M-phase of 9.4% and for S+G2/M-phase fractions of 15.04% as best predicting survival. The patients with glioma had poor survival when the levels of G2/M-phase and S+G2/M-phase were more than 9.4 and 15.04%, respectively. In contrast, no significant association between the DNA content of glioma patients and their age, tumor recurrence, and tumor size was found.</p> <p><b>Discussion:</b> Our results indicate that flow cytometry analysis for G2/M-phase and S+G2/M-phase fractions can be used for tumor grading for rapidly differentiating low- from high-grade gliomas.</p

Physiologic parameters before (preocclusion) and after (postocclusion) permanent middle cerebral artery occlusion (PMCAO) between animals pretreated with magnolol versus vehicle (PEG 400)-treated controls.

<p>Physiologic data obtained from control and pre-treated animal groups are represented as the mean±standard deviation (SD). Hct – hematocrit; Gluc - blood glucose; MABP - mean arterial blood pressure; HR - heart rate; <i>n</i> – number of animals. All animals were maintained at 37±0.5°C. Paired Students’ <i>t</i> tests were used to evaluate the response to a change in conditions, and one-way Analysis of Variance (one-way ANOVA) with Dunnett’s <i>posthoc</i> comparison was used to evaluate differences between groups. The symbol * and <b>†</b> mean <i>P<</i>0.05, compared to preischemic and control data, respectively.</p

Chemical Constituents and Anti-inflammatory Principles from the Fruits of <i>Forsythia suspensa</i>

Fifty compounds were isolated from the fruits of <i>Forsythia suspensa</i>, including 13 new compounds characterized as eight new diterpenoids (<b>1</b>–<b>8</b>), three new lignans (<b>9</b>–<b>11</b>), a new iridoid (<b>12</b>), and a new triterpenoid (<b>13</b>). Their structures were established on the basis of spectroscopic and spectrometric analysis. Most of the isolated compounds were examined for their anti-inflammatory activity in vitro. The results showed that several compounds displayed significant inhibition of fMLP/CB-induced superoxide anion generation and elastase release, with IC₅₀ values ranging from 0.6 ± 0.1 to 8.6 ± 0.8 μg/mL and from 0.8 ± 0.3 to 7.3 ± 1.1 μg/mL, respectively

Magnolol attenuated glutamate-induced rises in the intracellular calcium, [Ca²⁺](i), inflow and cell swelling in cultured neurons.

<p>(A) Ratio image detection for [Ca²⁺](i) concentrations showed that magnolol at 0.1 µM, but not at 0.01 and 1 µM, effectively inhibited the rises of [Ca²⁺](i) induced by glutamate exposure. (B) Time-course differential interference contrast (DIC) photomicrographs of cultured neurons showed that magnolol at 0.1–1 µM attenuated the glutamate-induced cell swelling over time. *<i>P</i><0.05 vs controls, and <i>n</i> = 6–7 per group.</p

Magnolol reduced glutamate-and N-methyl-D-aspartate (NMDA)-induced cell deaths.

<p>(A, B) Differential interference contrast (DIC) photomicrographs showed neurotoxicity of magnolol at a concentration beyond 100 µM. (C, D) Magnolol (0.1–1 µM) achieved potent cytoprotection against glutamate-induced neuronal damage. (E) Magnolol (0.1–1 µM) achieved potent cytoprotection against NMDA-induced neuronal damage. *<i>P</i><0.05 vs controls, and <i>n</i> = 5–9 per group.</p

Neurobehavioral scores and body weight loss obtained after permanent middle cerebral artery occlusion (pMCAO) in each pretreatment group in the study.

<p>Data are represented as the mean±standard deviation (S.D.). <i>n</i> – number of animals.</p>*<p>means <i>P</i><0.05, compared to control data, respectively.</p

The changes of core temperatures obtained after vehicle or magnolol treatment in rats subjected to permanent middle cerebral artery occlusion (PMCAO) in the study.

<p>Data are represented as the mean±standard deviation (S.D.). <i>n</i> – number of animals.</p>*<p>means <i>P</i><0.05, compared to control data, respectively.</p

Dragonbloodin A1 and A2: Flavan Trimers and Anti-inflammatory Principles from Sanguis Draconis

Two flavan trimers, dragonbloodin A1 (<b>1</b>) and A2 (<b>2</b>), were isolated as diastereomers from sanguis draconis, a traditional Chinese medicine for regulating blood. The structures of <b>1</b> and <b>2</b> were elucidated by spectroscopic analysis and X-ray diffraction. Possible interactions between <b>1</b> and <b>2</b> were discussed, and possible biosynthesis pathways were deduced. Compounds <b>1</b>, <b>2</b>, and their racemic mixture all exhibited inhibition of human neutrophil elastase in a dose-dependent manner

Anti-inflammatory Flavan-3-ol-dihydroretrochalcones from <i>Daemonorops draco</i>

Four A-type flavan-3-ol-dihydroretrochalcone dimers, dragonins A–D (<b>1</b>–<b>4</b>), were characterized from the traditional Chinese medicine Sanguis Draconis. The structures of <b>1</b>–<b>4</b> were elucidated by spectroscopic and spectrometric analyses. Compounds <b>1</b> and <b>2</b> exhibited significant inhibition of fMLP/CB-induced superoxide anion and elastase. The signaling pathways accounting for the inhibitory effects of compound <b>2</b> were also elucidated. These purified A-type flavan-3-ol-dihydroretrochalcones are new potential leads for the development of anti-inflammatory drugs

Dragonbloodin A1 and A2: Flavan Trimers and Anti-inflammatory Principles from Sanguis Draconis

Two flavan trimers, dragonbloodin A1 (<b>1</b>) and A2 (<b>2</b>), were isolated as diastereomers from sanguis draconis, a traditional Chinese medicine for regulating blood. The structures of <b>1</b> and <b>2</b> were elucidated by spectroscopic analysis and X-ray diffraction. Possible interactions between <b>1</b> and <b>2</b> were discussed, and possible biosynthesis pathways were deduced. Compounds <b>1</b>, <b>2</b>, and their racemic mixture all exhibited inhibition of human neutrophil elastase in a dose-dependent manner