HBB contributes to individualized aconitine-induced cardiotoxicity in mice via interfering with ABHD5/AMPK/HDAC4 axis.

The root of Aconitum carmichaelii Debx. (Fuzi) is an herbal medicine used in China that exerts significant efficacy in rescuing patients from severe diseases. A key toxic compound in Fuzi, aconitine (AC), could trigger unpredictable cardiotoxicities with high-individualization, thus hinders safe application of Fuzi. In this study we investigated the individual differences of AC-induced cardiotoxicities, the biomarkers and underlying mechanisms. Diversity Outbred (DO) mice were used as a genetically heterogeneous model for mimicking individualization clinically. The mice were orally administered AC (0.3, 0.6, 0.9 mg· kg-1 ·d-1) for 7 d. We found that AC-triggered cardiotoxicities in DO mice shared similar characteristics to those observed in clinic patients. Most importantly, significant individual differences were found in DO mice (variation coefficients: 34.08%-53.17%). RNA-sequencing in AC-tolerant and AC-sensitive mice revealed that hemoglobin subunit beta (HBB), a toxic-responsive protein in blood with 89% homology to human, was specifically enriched in AC-sensitive mice. Moreover, we found that HBB overexpression could significantly exacerbate AC-induced cardiotoxicity while HBB knockdown markedly attenuated cell death of cardiomyocytes. We revealed that AC could trigger hemolysis, and specifically bind to HBB in cell-free hemoglobin (cf-Hb), which could excessively promote NO scavenge and decrease cardioprotective S-nitrosylation. Meanwhile, AC bound to HBB enhanced the binding of HBB to ABHD5 and AMPK, which correspondingly decreased HDAC-NT generation and led to cardiomyocytes death. This study not only demonstrates HBB achievement a novel target of AC in blood, but provides the first clue for HBB as a novel biomarker in determining the individual differences of Fuzi-triggered cardiotoxicity.

Effects of tenuifolin on rest/wake behaviour in zebrafish.

Insomnia is a common sleep disorder with a high prevalence and substantial adverse consequences. There is growing interest in identifying novel therapeutics from herbal medicine. Tenuifolin is a major constituent of the well-known anti-insomnia herb Radix Polygala. The present study investigated the neural activity in response to tenuifolin during rest/wake behaviour in zebrafish and identified the potential biological signalling pathways involved. An automatic video tracking system was used to monitor the behavioural response of zebrafish larvae for 24 h after treatment with tenuifolin. In total, six rest/wake parameters were measured and visualized with a behavioural fingerprint. Time series analysis was conducted by averaging the total rest and waking activity in 10 min intervals. A correlation analysis was performed between tenuifolin and well-known compounds to analyse the underlying biological signalling pathways. Reverse transcription-quantitative PCR was also performed to detect the effects of tenuifolin on the transcription of interesting genes associated with the signalling pathways that were potentially involved. The present results suggested tenuifolin significantly increased the total rest time during the dark phase, with a slight effect on the waking activity in zebrafish larvae. This behavioural phenotype induced by tenuifolin is similar to that of selective serotonin reuptake inhibitors and gamma-aminobutyric acid (GABA) agonists. Furthermore, the expression levels of GABA transporter 1 were significantly increased after tenuifolin treatment. No significant difference was determined in other associated genes in untreated control and tenuifolin-treated larvae. The present results suggested that tenuifolin caused sleep-promoting activity in zebrafish and that these effects may be mediated by the serotoninergic systems and the GABAergic systems.

[Analysis on regularity of traditional Chinese patent medicine in treating hyperlipidemia].

To explore the regularity of traditional Chinese patent medicines for the treatment of hyperlipidemia recorded in Newly Edited National Chinese Traditional Patent Medicines,the Composition Principles of Chinese Patent Drugs,New Drug Conversion Standard,the Compilation of National Standard for Traditional Chinese Medicines and Chinese Pharmacopoeia. Researchers extracted the information of prescriptions from these cases according to the inclusion and exclusion criteria. Then microsoft excel 2010 was used to conduct frequency statistics and count the frequency of traditional Chinese medicine. SPSS Clementine( ver.12. 0) and SPSS( ver. 18. 0)were adopted respectively for frequency analysis,association rules analysis,cluster analysis and factor analysis. Besides,KMO test and Bartlett spherical test were performed for factor adaptation test. Finally,a total of 173 traditional Chinese medicines were included,involving 94 Chinese patent medicine prescriptions. The frequency results of traditional Chinese medicine showed that there were 33 kinds of high-frequency traditional Chinese medicine,mainly including those for tonifying medicine,activating blood and resolving stasis and blood-stasis,and clearing damp. The association rules analysis found out 12 association rules of drug pairs,3-herb pairs of 25 and4-herb pairs of 6. Totally 11 medicine groups with relevance were respectively extracted by cluster analysis. KMO test and Bartlett spherical test indicated that the method was suitable for factor analysis and 11 common factors were respectively extracted by factor analysis. The association rules reflected the therapeutic method for tonify the liver and kidney,activating blood and resolving stasis. Cluster analysis and factor analysis showed the therapeutic method of Qi-enriching and Yin-nourishing,and the factor analysis focused more on removing blood stasis and dampness. The decision tree with hawthorn as the dependent variable reflects the importance of alisma orientalis and fructus schisandrae in the drug matching. In conclusion,data mining technique can comprehensively analyze the regularity of prescriptions of traditional Chinese patent medicine for hyperlipidemia,and is helpful for guiding the development of Chinese patent medicines and the clinical practice of traditional Chinese medicine.

[Analysis of treating bi syndrome by I-Ching navel acupuncture].

Based on the theories of I-Ching and umbilicus-hologram, the navel acupuncture is considered as a new acupuncture therapy that only acupuncture at Shenque (CV 8). It has a good effect on the treatment of bi syndrome and provides a new treatment idea for bi syndrome. This article presents the definition, etiology and treatment of bi syndrome, and introduces the application of umbilical-holographic, the principle and method of positioning and needle-inserting, the adjustment of therapies and the analysis of cases, in order to introduce the idea of treating bi syndrome by I-Ching navel acupuncture.

[Study on estrogen receptor of adult human osteoblasts].

OBJECTIVE: To examine estrogen receptor (ER) in osteoblasts from adult human and to elucidate the mechanism of estrogen in modulating bone metabolism. METHODS: The cultured osteoblasts were harvested from bone chips by modified sequential digestive enzyme release and immunohistochemical assay of ER in osteoblasts were carried out in three groups of female adults: normal control (group 1), patients with moderate osteoporosis (group 2) and patients with serious osteoporosis (group 3). The percentages of ER-positive osteoblasts from the three groups were compared by t test. RESULTS: The brown marks that indicate ER were found in nuclei and plasma of the osteoblasts, and the percentages of ER-positive osteoblasts among three groups were significantly different. CONCLUSION: ERs exist in nuclei and plasma of the osteoblasts. Estrogen may modulate bone metabolism through binding ER in nuclei and plasma of the osteoblasts. The reduction of ER of osteoblasts may play an important role in the pathogenesis of postmenopausal osteoporosis.

[Culture of human osteoclasts].

OBJECTIVE: To establish a stable, useful culture system for human osteoclasts and to investigate the effect of osteoblasts on the differentiation, proliferation and activation of osteoclasts so as to provide a base for the studies on prevention and treatment of osteolysis and osteoporosis. METHODS: In the presence of 1,25-(OH)2D3, monocytes abstracted from human bone marrow were cultured in three groups: co-culture of monocytes and osteoblasts, monocytes alone, monocytes with conditional media (CM) of osteoblasts. Differentiation process of the cultured cells was observed under biological microscope. HE staining and tartrate-resistant acid phosphatase (Trap) staining were employed to assay the cultured cells. The resorption pits on bone slices, on which cells were cultured, were observed under scanning electronic microscope (SEM). RESULTS: In the group of co-culture of monocytes and osteoblasts, monocytes gradually fused to form multinucleated cells (MNCs), and the MNCs were also indicated in HE staining and Trap staining. The SEM showed a number of resorption pits on bone slices. In the other two groups, Trap-positive MNCs were not obtained, and resorption pits were not observed on bone slices. CONCLUSION: In this culture, monocytes obtained from human marrow fused to form multinucleated cells (MNCs) that express the main characteristics of the osteoclast phenotype, such as Trap-positive and the ability to form resorption lacunae when cultured on bone slices. Cell-to-cell contact with osteoblasts was necessary for the differentiation, proliferation and activation of osteoclasts.