AAV-Mediated nuclear localized PGC1α4 delivery in muscle ameliorates sarcopenia and aging-associated metabolic dysfunctions.

Sarcopenia is characterized of muscle mass loss and functional decline in elder individuals which severely affects human physical activity, metabolic homeostasis, and life quality. Physical exercise is considered effective in combating muscle atrophy and sarcopenia, yet it is not feasible to elders with limited mobility. PGC-1α4, a short isoform of PGC-1α, is strongly induced in muscle under resistance training, and promotes muscle hypertrophy. In the present study, we showed that the transcriptional levels and nuclear localization of PGC1α4 was reduced during aging, accompanied with muscle dystrophic morphology, and gene programs. We thus designed NLS-PGC1α4 and ectopically express it in myotubes to enhance PGC1α4 levels and maintain its location in nucleus. Indeed, NLS-PGC1α4 overexpression increased muscle sizes in myotubes. In addition, by utilizing AAV-NLS-PGC1α4 delivery into gastrocnemius muscle, we found that it could improve sarcopenia with grip strength, muscle weights, fiber size and molecular phenotypes, and alleviate age-associated adiposity, insulin resistance and hepatic steatosis, accompanied with altered gene signatures. Mechanistically, we demonstrated that NLS-PGC-1α4 improved insulin signaling and enhanced glucose uptake in skeletal muscle. Besides, via RNA-seq analysis, we identified myokines IGF1 and METRNL as potential targets of NLS-PGC-1α4 that possibly mediate the improvement of muscle and adipose tissue functionality and systemic energy metabolism in aged mice. Moreover, we found a negative correlation between PGC1α4 and age in human skeletal muscle. Together, our results revealed that NLS-PGC1α4 overexpression improves muscle physiology and systematic energy homeostasis during aging and suggested it as a potent therapeutic strategy against sarcopenia and aging-associated metabolic diseases.

Development of high-performance mixed matrix reverse osmosis membranes by incorporating aminosilane-modified hydrotalcite.

Thin film nanocomposite (TFN) reverse osmosis (RO) membranes were prepared by dispersing 3-aminopropyltriethoxysilane (APTES) modified hydrotalcite (HT), designated as A-HT, in aqueous solution and incorporating the nanoparticles in polyamide layers during the interfacial polymerization process. Results of Fourier transform infrared spectroscopy and zeta potential characterization showed the successful modification of nanoparticles by APTES. In addition, Fourier transform infrared spectroscopy suggested that amidation would take place between the aminosilane on APTES and trimesoyl chloride in organic solution, providing firm covalent interaction between the nanoparticles and polyamide matrix. Dynamic light scattering and transmission electron microscopy indicated that aminosilane modification improved dispersibility of the nanoparticles in aqueous solution and obtained membranes, which suppressed the aggregation. Both the covalent interaction and aggregation suppression were beneficial to compatibility between nanoparticles and the polyamide matrix. TFN RO membranes incorporated with A-HT demonstrated excellent performance. Compared with the pristine RO membrane, the water flux of A-HT-0.050 prepared with an optimum A-HT concentration of 0.050 wt% was enhanced by 18.6% without sacrificing the salt rejection. Moreover, the selectivity of A-HT-0.050 was superior to that of HT-0.050 prepared with HT of 0.050 wt%, which proved aminosilane modification of hydrotalcite was beneficial to high membrane performance especially to selectivity.

Evidence for heightened genetic instability in precancerous spasmolytic polypeptide expressing gastric glands.

BACKGROUND: Spasmolytic polypeptide-expressing metaplasia (SPEM) is present in more than 90% of resected gastric cancer tissues. However, although widely regarded as a pre-cancerous tissue, its genetic characteristics have not been well studied. METHODS: Immunohistochemistry using Trefoil factor 2 (TFF2) antibodies was used to identify TFF2-positive SPEM cells within SPEM glands in the stomach of Helicobacter felis (H. felis) -infected mice and human clinical samples. Laser microdissection was used to isolate specific cells from both the infected mice and the human samples. The genetic instability in these cells was examined by measuring the lengths of microsatellite (MS) markers using capillary electrophoresis. Also, genome-wide genetic variations in the SPEM cells from the clinical sample was examined using deep whole-exome sequencing. RESULTS: SPEM cells indeed exhibit not only heightened MS instability (MSI), but also genetic instabilities that extend genome-wide. Furthermore, surprisingly, we found that morphologically normal, TFF2-negative cells also contain a comparable degree of genomic instabilities as the co-resident SPEM cells within the SPEM glands. CONCLUSION: These results, for the first time, clearly establish elevated genetic instability as a critical property of SPEM glands, which may provide a greater possibility for malignant clone selection. More importantly, these results indicate that SPEM cells may not be the sole origin of carcinogenesis in the stomach and strongly suggest the common progenitor of these cells, the stem cells, as the source of these genetic instabilities, and thus, potential key players in carcinogenesis.

Single-cell gene variation analysis method for single gland.

Single-cell analysis of heterogeneity has become the cutting-edge technology for profound understandings of relationships between cell populations. At present, common methods used in single cellular genomic research are mainly microfluidic technologies (Fluidigm) or based on microwells, both requiring a uniform size of cells at the entrance. However, the size of cells in specific tissues can vary from type to type. To address this issue, we need to establish a method to identify genomic features of individual cells of different sizes. In this paper, we developed a robust method in the analysis of single cellular genomic mutations among gastric tissues. Briefly, the single gastric gland was isolated from the whole tissue, and further enzymatically digested into single cells of various sizes by trypsin. These single cells were then spread on the polyethylene naphthalene slides and selected by the laser microdissection method. Whole genome amplification (WGA) and capillary electrophoresis were performed subsequently to detect single cell microsatellite. This method enabled us to detect the existence of microsatellite instability (MSI) of each single cell within the intestinal metaplasia, and to carry out a flexible and fine analysis of single cells with different sizes in tissues and glands. This reliable and practical method is well performed in both low and high-throughput genome analysis when combined with cell labeling methods, thus providing a novel and highly flexible way to study tissue heterogeneity on the single cell scale.

Lipase immobilized catalytically active membrane for synthesis of lauryl stearate in a pervaporation membrane reactor.

A composite catalytically active membrane immobilized with Candida rugosa lipase has been prepared by immersion phase inversion technique for enzymatic synthesis of lauryl stearate in a pervaporation membrane reactor. SEM images showed that a "sandwich-like" membrane structure with a porous lipase-PVA catalytic layer uniformly coated on a polyvinyl alcohol (PVA)/polyethersulfone (PES) bilayer was obtained. Optimum conditions for lipase immobilization in the catalytic layer were determined. The membrane was proved to exhibit superior thermal stability, pH stability and reusability than free lipase under similar conditions. In the case of pervaporation coupled synthesis of lauryl stearate, benefited from in-situ water removal by the membrane, a conversion enhancement of approximately 40% was achieved in comparison to the equilibrium conversion obtained in batch reactors. In addition to conversion enhancement, it was also found that excess water removal by the catalytically active membrane appears to improve activity of the lipase immobilized.

Consequence alimentary reconstruction in nutritional status after total gastrectomy for gastric cancer.

AIM:To investigate the effect of gastroenteric reconstruction on the nutritional status of patients with gastric cancer after total gastrectomy.METHODS: From 1989-1994, nutritional status was studied in 24 patients, including 12 patients with the gastric reservoir and pyloric sphincter reconstruction (GRPS), 7 with Braun's esophago-jejunostomy (EJ) and 5 with Lawrance's Roux en Y reconstruction (RY).The ability of these patients to ingest and absorb the amount of nutrients was examined and compared, and metabolic balance test was performed to compare the efficiency of those patients to accumulate and use the absorbed nutrients.RESULTS:In the controlled hospital situation, the amount of food ingested by all the patients was greater than that required for maintenance of ideal body weight. In direct contrast, food intake in most patients with EJ or RY reconstruction significantly decreased when the patients returned home and that in EJ patients it was the lowest. The overgrowth of anaerobic bacteria was found in the jejunum in the patients with EJ and RY, due mainly to food stasis in the duodenum or in the Roux limb,caused by the operative procedure itself. In patients with GRPS,because of restoring of the alimentary continuity according to the normal digestive physiologic characters, all the nutritional parameters could fall in the normal range.CONCLUSION:The most common mechanism responsible for postoperative malnutrition was inadequate food intake. Having solved the problem of alkaline reflux esophagitis, it is imperative to preserve the duodenal food passage to reduce malabsorption and other complications after total gastrectomy.