Adaptive evolution analysis of squalene synthase in medicinal plants of Araliaceae / 中草药

Objective: To analyze the adaptive evolution of squalene synthase (SS) in the medicinal plants of Araliaceae. Methods: The adaptive evolutionary analysis of 23 SS genes of seven medicinal plants of Araliaceae was carried out by using the branch model, site model, branch-site model, MEC model, SLAC, FEL, and REL of PAML software. Results: In the analysis of PAML and MEC models, most of the branches and loci were found to be under strong negative selection, and no positive selection sites were found. The analysis by SLAC, FEL, and REL also showed that there were a large number of negative selection sites, only 412P, 413N, and 415K were positive selection sites. Conclusion: This indicates that negative selection plays a leading role in SS gene of Araliaceae. The 412P, 413N, and 415K sites found may be involved in the activity of SS.

El retículo endoplasmático liso en hepatocitos estimulados con distintas dosis de láser infrarrojo / The smooth endoplasmic reticulum in hepatocytes stimulated with different doses of infrared laser

Veinticuatro ratas hembras Sprague Dawley de 4 meses de vida con peso aproximado de 250 g, fueron divididas en cuatro grupos (A, B, C y D), donde el grupo A (control) no recibió estimulación infrarroja, B se irradió con láser infrarrojo 4 J/cm², C con dosis de 8 J/cm² y D con 16 J/cm². La estimulación infrarroja se realizó diariamente, por 15 días ininterrumpidos. Las ratas fueron sacrificadas y se extrajeron muestras tanto de hígado normal (control) como estimulado con las distintas dosis infrarrojas, las que fueron procesadas para microscopía electrónica de transmisión. De los hepatocitos normales y estimulados, se obtuvieron microfotografías con aumentos finales de hasta 36.500 X, que fueron sometidas a estudios morfométricos para determinar fracciones volumétricas con especial énfasis en el retículo endoplásmico liso (REL) y de los siguientes componentes celulares: retículo endoplasmático rugoso (RER), mitocondrias, glicógeno, eu y heterocromatina. De igual manera se cuantificaron las áreas celulares y nucleares. Del análisis de los resultados entre hepatocitos normales y estimulados con diferentes dosis infrarrojas, se visualiza que existen notables diferencias en todos los componentes celulares cuantificados particularmente el REL. Se concluye que las estimulaciones infrarrojas provocan una drástica transformación en la ultraestructura y morfología de los hepatocitos, lo que provocaría una variación funcional, representando de esta manera el efecto que estas estimulaciones provocan en este tipo celular.

A total of 24 female Sprague-Dawley rats aged 4 months and weighing approximately 250 g, were divided into four groups labeled A, B, C and D. Group A received no infrared stimulation and served as control. Group B was radiated with a dose of 4 J/cm² of infrared laser, Group C with doses of 8 J/cm² and Group D with 16 J/cm². This infrared stimulation was carried out daily for 15 days uninterrupted. The rats were then sacrificed and samples of both normal-control liver and liver stimulated with the different infrared doses were extracted for immediate processing via transmission electron microscopy. Transmission electron microphotographs were obtained at magnifications of 21300X from both normal and stimulated hepatocytes; these were subjected to morphometric studies to determine volumetric fractions with special emphasis on the smooth endoplasmic reticulum (SER) and the following cell components: rough endoplasmic reticulum (RER), mitochondria, glycogen, eu and heterochromatin. Likewise, cell and nuclear areas were quantified. Analysis of the results of normal and stimulated hepatocytes with different infrared doses showed considerable differences in all the quantified cell components and particularly from the SER it is concluded that the effects of these stimulations bring about a drastic transformation in the ultrastructure and morphology of the hepatocytes, which may ultimately translate into a functional variation, thus representing the effect that these stimulations cause in this cell type.

Expression of NF-κB subunits P50 and c-Rel protein in primary cortical neurons after oxygen glucose deprivation/reoxygenation / 第二军医大学学报

Objective: To investigate the expression of nuclear factor-κB (NF-κB) subunits P50 and c-Rel protein in primary cortical neurons of Wistar rats at different time points of oxygen glucose deprivation/ reoxygenation(OGD/R). Methods: The neurons dissociated from the cortex of the neonatal rats were primary cultured and were identified by immunocytochemistry. OGD/R model was established. The study was divided into 6 groups according to different processing methods,including normal group,OGD 4 h treated,OGD 4 h/R 2 h treated,OGD 4 h/R 6 h treated,OGD 4 h/R 12 h treated and OGD 4 h/R 24 h treated groups. The expression of NF-κB P50 and c-Rel protein in neurons was examined by immunocytochemistry method and Western blotting. Results: (1) Immunocytochemistry detection targeting neuron specific enolase (NSE) and beta-III tubulin confirmed that the cultured cells were neurons. (2) The expression of NF-κB P50 protein was significantly higher in OGD 4 h group than in control group(P<0.05); the expression continued to increase in OGD 4 h/R 2 h andOGD 4 h/R 6 h groups, and reached its peak 6 h after reoxygenation (P<0.01),then began to decrease,but the expression in OGD 4 h/R 12 h group was significantly different from that of the control group (P<0.05); there was no significant difference between OGD 4 h/R 24 h group and control group. (3) The expression of NF-κB c-Rel protein was similar between OGD 4 h group and the control group; the expression increased in OGD 4 h/R 2 h and OGD 4 h/R 12 h groups and reached its peak 12 h after reoxygenation(P<0.01),and did not recover to the normal level in OGD 4 h/ R 24 h group (P<0.05). Conclusion: Oxygen glucose deprivation/reoxygenation can activate NF-κB subunits P50 and c-Rel in the primary cortical neurons in rats in a time-associated manner.

Magnolol Inhibits LPS-induced NF-kappaB/Rel Activation by Blocking p38 Kinase in Murine Macrophages

This study demonstrates the ability of magnolol, a hydroxylated biphenyl compound isolated from Magnolia officinalis, to inhibit LPS-induced expression of iNOS gene and activation of NF-kappaB/Rel in RAW 264.7 cells. Immunohisto-chemical staining of iNOS and Western blot analysis showed magnolol to inhibit iNOS gene expression. Reporter gene assay and electrophoretic mobility shift assay showed that magnolol inhibited NF-kappaB/Rel transcriptional activation and DNA binding, respectively. Since p38 is important in the regulation of iNOS gene expression, we investigated the possibility that magnolol to target p38 for its anti-inflammatory effects. A molecular modeling study proposed a binding position for magnolol that targets the ATP binding site of p38 kinase (3GC7). Direct interaction of magnolol and p38 was further confirmed by pull down assay using magnolol conjugated to Sepharose 4B beads. The specific p38 inhibitor SB203580 abrogated the LPS-induced NF-kappaB/Rel activation, whereas the selective MEK-1 inhibitor PD98059 did not affect the NF-kappaB/Rel. Collectively, the results of the series of experiments indicate that magnolol inhibits iNOS gene expression by blocking NF-kappaB/Rel and p38 kinase signaling.

Preparation and in Vitro Release Behavior of Paclitaxel Microsphere / 中国药房

OBJECTIVE:To optimize the formulation parameters of poly1,3-bis(p-carboxyphenoxy) propane-sebacic acid (P(CPP∶SA)) microsphere and evaluate its drug release performance in vitro. METHODS: The paclitaxel microspheres were prepared by single emulsion method,and the effects of the factors such as the stirring speed (A),P(CPP∶SA) concentration (B),emulsifier polyvinyl alcohol (PVA) concentration (C) on encapsulation efficiency were evaluated by orthogonal test. The surface morphology of the prepared microsphere was observed and its drug release performance in vitro was evaluated. RESULTS: The optimal formulation of the microsphere obtained was as follows: A was 4 000 r?min-1,B was 80 mg?mL-1 and C was 1%,respectively. The morphology of microspheres was round and intact. The encapsulation efficiency of paclitaxel was up to above 90%. The sustained release duration was 30 days with an accumulative release rate of over 80%. CONCLUSION: Paclitaxel microspheres prepared by optimal formula display a high encapsulation efficiency and satisfactory sustained-release pattern.

Inhibition of p65 Nuclear Translocation by KIOM-79 / 대한해부학회지

We demonstrate that KIOM-79, combined extracts isolated from Magnolia officinalis, Pueraria lobata, Glycyrrhiza uralensis, and Euphorbia pekinensis, inhibits LPS-induced expression of iNOS gene in RAW 264.7 cells. Treatment of RAW 264.7 cells with KIOM-79 inhibited LPS-stimulated nitric oxide production in a doserelated manner. Immunohisto-chemical staining of iNOS and RT-PCR analysis showed that the decrease of NO was due to the inhibition of iNOS gene expression. Immunostaining of p65 and EMSA showed that KIOM-79 inhibited NF-kappa/Rel nuclear translocation and DNA binding, respectively. Collectively, this series of experiments indicates that KIOM inhibits iNOS gene expression by blocking NF-kappa/Rel. Due to the critical role that NO release plays in mediating inflammatory responses, the inhibitory effects of KIOM-79 on iNOS suggest that KIOM-79 may represent a useful anti-inflammatory agent.

Nuclear Translocation of NF-kappaB/Rel Component by PCSC22 Isolated from Poria cocos Sclerotium / 대한해부학회지

The sclerotium of Poria cocos Wolf, which grows on the roots of pine trees, has long been used as a sedative and diuretic (Chang and But, 1987). The accumulating data revealed that certain ingredients of the sclerotium of Poria cocos showed anti-tumor activities (Kanayama, 1986). Although the mechanism of anti-tumor activity is not known, the polysaccharides may potentiate the host defense mechanism through the activation of immune system. In the present study we show that PCSC22, a polysaccharide isolated from the sclerotium of Poria cocos with one percent sodium carbonate, significantly induces nitric oxide (NO) production and inducible NO synthase (iNOS) transcription. To further investigate the mechanism responsible for the induction of iNOS gene expression, we investigated the effect of PCSC22 on the activation of NF-kappaB/Rel, whose binding site was located in the promoter of iNOS gene. Immuno-histo-chemical staining of p65 and p50 showed that PCSC22 produced strong induction of NF-kappaB/Rel nuclear translocation. Electrophoretic mobility shift assay (EMSA) further confirmed the activation of NF-kappaB/Rel by PCSC22. In conclusion, we demonstrate that PCSC22 stimulates macrophages to express iNOS gene through the activation of NF-kappa B/Rel.

Relationship Between Nuclear Factor-Kappa B and Restenosis after Angioplasty / 中国普外基础与临床杂志

Objective To discuss the role of nuclear factor kappa B in restenosis after angioplasty.Methods Related literatures of recent 5 years were reviewed.Results Nuclear factor kappa B could lead to hyperplasia of vascular intima which resulted from proliferation and decrease of apoptosis of vascular smooth muscle cells.Conclusion Nuclear factor kappa B plays an important role in restenosis after angioplasty.

Expression of NF-?B subunits P50 and c-Rel protein in primary cortical neurons after oxygen glucose deprivation/reoxygenation / 第二军医大学学报

Objective:To investigate the expression of nuclear factor-?B(NF-?B)subunits P50 and c-Rel protein in primary cortical neurons of Wistar rats at different time points of oxygen glucose deprivation/reoxygenation(OGD/R).Methods:The neurons dissociated from the cortex of the neonatal rats were primary cultured and were identified by immunocytochemistry.OGD/R model was established.The study was divided into 6 groups according to different processing methods,including normal group,OGD 4 h treated,OGD 4 h/R 2 h treated,OGD 4 h/R 6 h treated,OGD 4 h/R 12 h treated and OGD 4 h/R 24 h treated groups.The expression of NF-?B P50 and c-Rel protein in neurons was examined by immunocytochemistry method and Western blotting.Results:(1)Immunocytochemistry detection targeting neuron specific enolase(NSE)and beta-Ⅲ tubulin confirmed that the cultured cells were neurons.(2)The expression of NF-?B P50 protein was significantly higher in OGD 4 h group than in control group(P