The hemolymph-like HL3 saline(Stewart et al., 1994)and standard saline(Jan & Jan, 1976)are two widely used bathing solutions for physiological recordings at the Drosophila larval neuromuscular junction. It has been established that longevity of larval preparations is better maintained in HL3 saline. However, HL3 can produce results that are inconsistent with previous findings in standard saline, particularly on temperature sensitivity and membrane excitability phenotypes. In wild-type larvae, the excitatory junctional potentials(EJPs)in standard saline(containing 4 mM Mg(2+)and 1.8 mM Ca(2+))were not blocked by a temperature increase up to 39-40 degrees C, consistent with unimpaired larval locomotion below these temperatures. However, in HL3 saline(containing 20 mM Mg(2+)and 1.5 mM Ca(2+)), EJPs were blocked at 30 degrees C. As for temperature-sensitive mutants nap(ts)and para(ts), the EJP-blocking temperatures were decreased from about 29 and 33 degrees C in standard saline to about 23 and 26 degrees C in HL3, respectively. Compound action potential recordings confirmed that segmental nerve action potentials were more readily blocked by a temperature increase in HL3 than in standard saline. Axonal excitability was suppressed in HL3 even at room temperatures, as evidenced by a lengthened refractory period in wild-type larvae. Similar suppression occurred for the hyper-excitable double mutant eag Sh, which maintained high-frequency spontaneous EJPs in standard saline but showed a rapidly declining EJP frequency in HL3. Application of HL3 saline also strongly suppressed the prolonged transmitter release following removal of repolarization mechanisms by K(+)channel blockers or by the eag Sh mutation previously described in standard saline. These discrepancies suggest that the high divalent cation content in HL3 may confer a surface charge screening effect to suppress nerve membrane excitability. We found that a minimal adjustment of the HL3 saline, decreasing the Mg(2+)ion concentration from 20 to 4 mM, was sufficient to resolve the discrepancies. While retaining the longevity of the larval neuromuscular preparation, the modified HL3 saline(HL3.1)restored the established wild-type EJP properties as well as phenotypes of several widely used temperature-sensitive and hyper-excitable mutants previously documented in standard saline.

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Although regenerated silk fibroin (SF), which has excellent biocompatibility, biodegradability, and a low inflammatory response in vivo, has promising applications in tissue engineering, the mechanical properties and biofunctionality must be further improved to satisfy tissue-engineering applications. Cellulose nanofibrils (CNFs) are promising candidates for bionanocomposite production due to their ultrahigh strength and excellent biocompatibility. In this study, CNFs were extracted directly from microcrystalline cellulose using an aqueous lithium bromide solution, a typical solvent for dissolving SF fibers. As a result, SF/cellulose nanocomposite films with improved tensile strength were fabricated using aqueous lithium bromide solution as a novel solvent system for the dissolution and blending of SF and cellulose. The extracted CNFs were homogeneously dispersed within the composite films through the rapid gelation of cellulose. The degradability of the composite films in a protease XIV solution was strongly dependent upon the SF component, which significantly promoted the degradation rate of composite films. Adhesion and proliferation results showed that SF/cellulose nanocomposite films promoted cell viability. Our work suggests a facile and effective approach for designing SF/cellulose nanocomposites that may have wide potential applications in tissue engineering.

Renal cell carcinoma (RCC) is one of the most common kidney cancers worldwide. Although great progressions have been made in the past decades, its morbidity and lethality remain increasing. Long noncoding RNAs (lncRNAs) are demonstrated to play significant roles in the tumorigenesis. This study aimed to investigate the detailed roles of lncRNA FTX in RCC cell proliferation and metastasis. Our results showed that the transcript levels of FTX in both clinical RCC tissues and the cultured RCC cells were significantly upregulated and associated with multiple clinical parameters of RCC patients, including familial status, tumor sizes, lymphatic metastasis, and TNM stages. With cell proliferation assays, colony formation assays, and cell cycle assays, we testified that knockdown of FTX in A498 and ACHIN cells with specific shRNAs inhibited cell proliferation rate, colony formation ability, and arrested cell cycle in the G0/G1 phase. FTX depletion also suppressed cell migration and invasion with Transwell assays and wound-healing assays. These data indicated the pro-oncogenic potential of FTX in RCC, which makes it a latent therapeutic target of RCC diagnosis and treatment in the clinic.