[Effects of Bushen Zhuanggu Granules on expressions of serum GH with IGF-1 and its receptors in bone tissue of ovariectomized rats].

Objective: To investigate the effects of Bushen Zhuanggu granule on the expressions of serum growth hormone (GH) and insulin-like growth factor-1(IGF-1) and their receptors in bone tissues of ovariectomized rats. Methods: Forty-eight SD female rats (weight 273±21.3 g) were divided into 4 four groups: the dosage of Bushen Zhuanggu granule group (BSZG) was 2.5 g/(kg·d),the do -sage of estradiol group(E2) was 0.071mg/(kg·d),sham group (SHAM) and ovariectomized model group (OVX group) were given the same amount of saline by oral administration.Each group included 12 rats,the treatments were conducted once a day. After 3 and 6 months of treatment,bone mineral density (B -MD) was measured by bone density instrument;the serum levels of GH and IGF-1 were detected by EL-ISA;the expressions of GHR and IGF-1R of bone tissue were detected by qPCR;the optical density(OD)value and positive cell count of pituitary GH immunohistochemical tablets were analyzed by Image J software,respectively. Results: ①After 3 months intervention,compared with the SHAM, the BMD of the spine in E2 group was increased(P＜0.05),and the BMD of both parts in BSZG group were increased(P＜0.05).After two stage intervention,BMD of both part in the two drug groups was higher than that in the OVX group(P＜0.05).②After two-stage intervention,the expression levels of serum GH and IGF-1,GHR and IGF-1R in BSZG group were higher than those in OVX group (P＜0.05).The levels of serum GH and it's receptors in the E2 group were increased (P＜0.05), but the serum IGF-1 level remained unchanged (P＞0.05) or even was decreased (P＜0.05).③After two stage in -tervention,the OD values and the positive cells count in the two drug intervention groups were increased (P＞0.05).④Pearson correlation analysis showed that serum GH,IGF-1 concentration and it's receptors in bone tissue were positively related with BMD. Serum GH concentration was positively correlated with OD values and number of positive cells. Conclusion: Bushen Zhuanggu granule can be used to improve the expressions of GH,IGF-1 in serum and its receptors in bone tissues of ovariectomized osteoporosis rats to prevent further loss of bone mass and increase bone mineral density.

An observation chamber for studying temperature-dependent and drug-induced events in live neurons using fluorescence microscopy.

Live-cell imaging chambers are used in a wide range of cell biology research. Recently, chambers capable of taking high-resolution and time-lapse images of live cells have been developed and become commercially available. However, because most of these chambers are designed to maintain a thermally stable environment for the cells under study, it is usually very difficult to use them to study temperature-dependent cellular events. Here we report the development of a chamber that is able to be used for the continuous monitoring of live neurons under most commercially available upright epifluorescence and confocal microscopes and in which the temperature and composition of the medium surrounding the neurons can be changed rapidly and reversibly. This live-cell observation chamber has been used successfully with cultured rat hippocampal neurons to study temperature-dependent changes in the dynamics of the microtubule cytoskeleton using fluorescence recovery after photobleaching (FRAP) together with the localization of alpha-tubulin in the dendritic spines. The success of these observations demonstrates the usefulness and applicability of the live-cell observation chamber described here to a wide range of cell biology experiments.

Studying the protein-protein interactions in the postsynaptic density by means of immunoabsorption and chemical crosslinking.

Postsynaptic densities (PSDs), isolated from porcine cerebral cortices, are large disk-shaped aggregates consisting of hundreds of different proteins. To study the protein-protein interactions in such complex supramolecules, we developed a procedure to break up the PSD's overall structure, while preserving some interactions between individual proteins. Using the resulting PSD sample and an indirect immunoabsorption procedure, PSD-95 was isolated along with the α- and ß-subunits of calcium calmodulin-dependent protein kinase II (CaMKIIα and CaMKIIß), α-tubulin, ß-tubulin, and Chapsyn110. Similarly, CaMKIIα was isolated along with CaMKIIß, α-tubulin, ß-tubulin, and small amounts of PSD-95. The proteins isolated from PSDs treated with a cleavable bifunctional crosslinking reagent were further subjected to diagonal gel electrophoresis analysis, and the results indicated that CaMKIIα resides next to α-tubulin in the PSD. Overall, the results obtained here suggest that within the PSD, large aggregates of CaMKIIα, CaMKIIß, α-tubulin, and ß-tubulin may occur that indirectly associate with PSD-95 and Chapsyn110. Such a protein organization would allow interactions with F-actin in the cytoplasm and with proteins, such as N-methyl-D-aspartate receptors, which reside on the postsynaptic membrane. Furthermore, it would facilitate binding to proteins such as the various microtubule-associated proteins that reside in the core region of the PSD.