Chicken TAX1BP1 suppresses type I interferon production via degrading chicken MAVS and facilitates infectious bursal diseases virus replication.

Mammalian TAX1BP1 (TAX1 binding protein 1), originally identified as a partner of the HTLV-1 viral oncoprotein, functions in regulation of cellular cytokine production. TAX1BP1 plays an important signal transduction regulator, specifically modulating innate immune signaling pathways including NF-B and IRF3. The function of TAX1BP1, which regulates the innate immune response in mammals, has been well studied in previous reports, but the role of chicken TAX1BP1 (chTAX1) in IFN regulation and infectious bursal disease virus (IBDV) replication is still unclear. In this report, chTAX1 was successfully cloned and sub-inserted into a eukaryotic expression vector. The critical regions of chTAX1, such as LC3 binding motif, ubiquitin binding motif, are highly conserved compared to other organisms. We also found that chTAX1 inhibits IFN expression by promoting degradation of chicken MAVS (chMAVS). In addition, the distribution of chTAX1 altered and translocated to co-localize with both VP1 and VP3 after IBDV infection. Overexpression of chTAX1 promotes IBDV replication and knockdown of chTAX1 by RNA interference suppresses IBDV replication. In summary, our data initially indicate that chTAX1 is a suppressor of IFN expression as well as a promoter of IBDV replication.

Preparation and evaluation of itraconazole dihydrochloride for the solubility and dissolution rate enhancement.

The purpose of this work was to explore the feasibility of preparing itraconazole hydrochloride to improve the solubility and dissolution rate. Itraconazole dihydrochloride was synthesized by bubbling anhydrous hydrogen chloride gas into the acetone suspension of itraconazole. Results of the elementary analysis gave the molecular formula of C(35)H(38)Cl(2)N(8)O(4).2HCl and its structure was confirmed by Fourier transform infrared (FTIR), thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC). Powder X-Ray diffraction (PXRD) suggested that a new crystalline form of the salt was formed. The morphology and mean size distribution study by scanning electron microscopy (SEM) and dynamic light scattering (DLS) confirmed that the salt was dispersable nanoparticle aggregation. Aqueous solubility measurements showed that the solubility of the salt, its 1:1, 1:2 and 1:3 (w/w) physical mixtures with beta-cyclodextrin (beta-CD) was 6, 99, 236 and 388 times greater than itraconazole. More than 94% of itraconazole was dissolved out of the salt/beta-CD 1/3 physical mixture after 60min. The stability studies indicated that the physical mixture remained stable for 24 months in assay, the related substances and dissolution. Based on the present results, it is concluded that hydrochloride formation can significantly increase solubility and dissolution rate of itraconazole, and the formulation of itraconazole dihydrochloride/beta-CD (1/3) would be an environment-friendly, economic and practical alternative to the commercially available itraconazole capsules (Sporanox)

[Image processing applying in analysis of motion features of cultured cardiac myocyte in rat].

Study of mechanism of medicine actions, by quantitative analysis of cultured cardiac myocyte, is one of the cutting edge researches in myocyte dynamics and molecular biology. The characteristics of cardiac myocyte auto-beating without external stimulation make the research sense. Research of the morphology and cardiac myocyte motion using image analysis can reveal the fundamental mechanism of medical actions, increase the accuracy of medicine filtering, and design the optimal formula of medicine for best medical treatments. A system of hardware and software has been built with complete sets of functions including living cardiac myocyte image acquisition, image processing, motion image analysis, and image recognition. In this paper, theories and approaches are introduced for analysis of living cardiac myocyte motion images and implementing quantitative analysis of cardiac myocyte features. A motion estimation algorithm is used for motion vector detection of particular points and amplitude and frequency detection of a cardiac myocyte. Beatings of cardiac myocytes are sometimes very small. In such case, it is difficult to detect the motion vectors from the particular points in a time sequence of images. For this reason, an image correlation theory is employed to detect the beating frequencies. Active contour algorithm in terms of energy function is proposed to approximate the boundary and detect the changes of edge of myocyte.

[Effect of melatonin and/or diazepam on circadian rhythm of cultured myocardiocytes in rats].

OBJECTIVE: To investigate the effect of melatonin and/or diazepam on the endogenous rhythm in cultured myocardiocytes and in that connexion to inquire into the mechanism of circadian oscillation level. METHODS: The beating rate of cultured myocardiocytes was dynamically monitored by an inverted phase-contrast microscope coupled to high speed CCD camera and computer system. Under the culturing condition of artificial light L:D = 12:12, the circadian rhythm variation of cultured myocardiocytes beating frequencies was measured, and then the effect exerted by melatonin and/or diazepam on that circadian rhythm was studied. RESULTS: It was found that under the L:D = 12:12 culturing condition, the cultured myocardiocytes exhibited prominent circadian rhythm. With the use of melatonin and or diazepam, the circadian rhythm of those myocardiocytes changed, not only the amplitude and mesor decreased, but also the acrophase delayed. CONCLUSION: The modulation of circadian rhythm in cellular level is not only related with endogenous control but also affected by external factors.

[Circadian expression of mPER1 in cultured murine myocardiocytes and effects of melatonin on it].

Objective. To investigate the mechanism of beating in cultured myocardiocytes through analyzing mPER1 expression and effect of melatonin on it. Method. Immunohistochemistry and melatonin interference test were employed. Result. mPER1 expression in cultured myocardiocytes showed circadian pattern, its acrophase was 15:20, its three consecutive daily average period length was approximately 23 h. Melatonin had little effects on its amplitude and period, but results in its phase delayed. The observation in this study was similar to those that we previously observed in cultured murine myocardiocytes beating. Conclusion. Oscillation of mPER1 gene is one of the important reasons which cause murine myocardiocytes circadian beating. Melatonin acts as "Zeitgeber" regulating mPER1 gene expression.