Rapid Purification of dsRNA Using Micro-Spin Cellulose Column.

Double-stranded RNA (dsRNA) is the replication intermediates of all RNA viruses. Purification and analysis of the profile and sequence of dsRNA is vital in virus diagnoses and/or characterization. Cellulose is one of the common materials used for isolation of dsRNA. Cellulose specifically binds dsRNA fraction under 15% ethanol concentration, which allows to isolate dsRNA from total nucleic acid solution or cell lysate. Here, we describe a rapid and reliable method for purifying dsRNA using a home-made micro-spin cellulose column from the cell lysate of virus-infected plant tissue. This labor-saving and rapid method enables routinely high-throughput isolation and analysis of dsRNA in plant or fungi samples.

Isolation of dsRNA from Plants by Cellulose Chromatography.

As the constitutive molecules of double-stranded RNA (dsRNA) virus genomes and replicative intermediates of single-stranded RNA (ssRNA) viruses, the high-molecular-weight dsRNAs are commonly found in RNA virus-infected plants. Therefore, the dsRNA is recognized as a hallmark of RNA virus infection and the profile of dsRNA has been applied as an efficient tool for diagnoses or characterization of unreported RNA viruses. Cellulose chromatography is one of the most useful procedures for the isolation of viral dsRNAs from total nucleic acids. Here, we describe rapid cellulose-based methods for purification of dsRNAs from plant tissue.

Study of optical parameters of polystyrene spheres in dense aqueous suspensions.

We investigated the dependence of the scattering and absorption coefficients of particles in dense suspensions by the low-coherence fiber optic dynamic light scattering (FODLS) technique. The estimated particle size was used to calculate the scattering coefficient of particles suspended in dense suspensions. The path-length resolved intensity distributions of light backscattered from absorbing dense suspensions were investigated experimentally. The absorption coefficient can be obtained by applying the measured path-length resolved intensity distributions to the modified Lambert-Beer law. As a result, the low-coherence FODLS technique can simultaneously measure the scattering and absorption coefficients of particles in absorbing dense suspensions, and the scattering and absorption coefficients are independent of each other in dense suspensions in the low-scattering regime of 2l(d) < 10ℓ*.

Clinical Evaluation of Titanium-Alloy Bands for Sternal Closure after Median Sternotomy: Single-Center Experience.

Background: Median sternotomy is the most applied approach in open-heart surgery, while potential complications such as postoperative bleeding, sternal dehiscence, and deep sternal wound infection (DWSI) still remain a challenge to cardiac surgeons. Several new sternum-closure products instead of stainless wire have been brought into clinical application. The objective of this retrospective study is to evaluate the novel sternum-fixing product in terms of clinical outcomes. Methods: 689 consecutive cases undergoing cardiac surgery through median sternotomy between February 2015 and December 2018 in our center were enrolled in this study. All the cases were divided into two groups according to different sternum fixation methods: wire cerclage group and rigid fixator group. The demographic as well as clinical data including the mediastinal drainage of first, second, and third post-op 24 hours, the total mediastinal drainage of post-op 72 hours, ICU duration, length of hospital stay, and post-op mortality in 30 days were collected and compared between the two groups. Results: 278 cases were enrolled in the wire cerclage group and 411 cases in the rigid fixator group. There is no significant difference in the demographic data between the two groups, while the mediastinal drainage in the first and third 24 hours after surgery and the total mediastinal drainage in postoperative 72 hours of the rigid fixator group were significantly less than those of the wire cerclage group (P < 0.05). No significant difference was found in other clinical outcomes between the groups including ICU duration, LOS in hospital, and 30-day mortality. 14 cases (5.0%) in the wire cerclage group and 11 cases (2.7%) in the rigid fixator group had sternotomy-related complications including severe postoperative bleeding, sternal dehiscence, and DSWI. Conclusion: Compared with the conventional wire cerclage, the new rigid fixator is superior in median sternotomy closure in terms of postoperative mediastinal bleeding as well as incidence of sternotomy-related complications.

Iridium(III) complexes entrapped in liposomes trigger mitochondria-mediated apoptosis and GSDME-mediated pyroptosis.

In this report, a new ligand TFBIP (TFBIP = 2-(4'-trifluoromethyl)-[1,1'-biphenyl]-4-yl)-1H-imidazo[4,5-f][1,10]phenanthroline) and its three iridium (III) complexes [Ir(ppy)2(TFBIP)](PF6) (Ir1, ppy = 2-phenylpyridine), [Ir(bzq)2(TFBIP)](PF6) (Ir2, bzq = benzo[h]quinolone) and [Ir(piq)2(TFBIP)](PF6) (Ir3, piq = 1-phenylisoquinoline) were synthesized and characterized. The cytotoxicity in vitro of the complexes toward several cancer cells was evaluated by 3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide (MTT) methods. The complexes show no cytotoxicity (IC50 > 100 µM) against these cancer cells. To enhance anticancer activity, these complexes were trapped in liposomes to form Ir1Lipo, Ir2Lipo and Ir3Lipo. The liposomes Ir1Lipo, Ir2Lipo and Ir3Lipo exhibit high or moderate cytotoxic activity. In particular, Ir1Lipo can effectively inhibit the cell growth with a low IC50 value (< 10 µM) toward A549, HepG2, BEL-7402, B16, HeLa and SGC-7901 cells. Surprisingly, Ir1Lipo has no cytotoxic activity against the normal cell LO2 (IC50 > 100 µM). The apoptosis and pyroptosis were investigated. Ir3Lipo induces apoptosis with a high early apoptotic number of 37%. The reactive oxygen species (ROS) levels, mitochondrial permeability transition pore open and mitochondrial membrane potential were detected. The intracellular Ca2+ concentration and release of cytochrome c were investigated. The expression of Bcl-2 (B-cell lymphoma-2) family proteins was explored by western blot. The antitumor activity in vivo of Ir1Lipo was evaluated with an inhibitory rate of 53%.

Metagenomic Analysis Reveals Neisseria bacilliformis Variation in the Early Childhood Caries Plaque Microbiome.

The progression of early childhood caries (ECC) is caused by microbial colonized in dental plaque. However, the association framework both from 16s genus down to high resolution metagenomic strain level and from composition to genome function analysis on caries lacks. 16S rRNA sequence revealed the composition of 3-6 years dental caries (ECC, n = 29), and severe dental caries (SECC, n = 36) children are significantly different from caries-free controls (CF, n = 31). Especially, genus Neisseria is enriched in caries (P < 0.05). Metagenomics sequence of 3 ECCs, 3 SECCs, and 3 CFs reveals Neisseria bacilliformis ATCC BAA-1200 in genus Neisseria is also significantly enriched in caries (P < 0.05). Then, we recovered high-quality metagenomic assembly genomes (MAG), named bin 86, which have 99% identity with Neisseria bacilliformis ATCC BAA-1200 genome. Function analysis of Neisseria bacilliformis ATCC BAA-1200 genome shows its metabolism power of sugar and adhesion, colonization, acid production, and acid tolerance ability, which suggested Neisseria bacilliformis ATCC BAA-1200 may serve as a biomarker for childhood caries.

Hematopoietic stem and progenitor cells can be enriched by implanting biomaterial into spatium intermusculare.

Hematopoietic stem and progenitor cells (HSPCs) have been used successfully to treat patients with cancer and disorders of the blood and immune systems. In this study, we tried to enrich HSPCs by implanting biomaterials into the spatium intermusculare of mice hind limbs. Gelatine sponges were implanted into the spatium intermusculare of mice and then retrieved after 12 days. The presence of HSPCs in the migrating cells (MCs) was detected by phenotypically probing with CD34(+)Sca-1(+) and functionally confirming the presence of using colony-forming cell assay and assessing the long-term reconstitution ability. The frequency of CD34(+), Sca-1(+), and CD34(+)Sca-1(+) cells and colony formation unit in the MCs was much higher than that in the bone marrow (BM). Moreover, transplanted MCs were able to home to BM, muscle, and spleen, which induced an efficient long-term hematopoietic reconstitution in vivo. In addition, HSPCs within the MCs originated from the BM. Furthermore, the administration of G-CSF greatly reduced the time of implantation, and increased the number of MCs and frequency of HSPCs in the MCs. These data provide compelling evidence that HSPCs can be enriched by implanting biomaterial into spatium intermusculare. Implantation of biomaterial may be seen as the first step to a proof of their applicability to clinical practice in enriching HSPCs.

Implantation of biomaterial as an efficient method to harvest mesenchymal stem cells.

Autologous mesenchymal stem cell (MSC) transplants have been used successfully to treat a number of diseases, and patients undergoing cell transplantation must have stem cells collected before transplantation. In this study, we developed a new method to harvest MSCs. Biomaterials were implanted into the spatium intermuscular of mice hind limbs, and a large number of migrating cells (MCs) were isolated from the transplanted biomaterials. The adherent cells in MCs showed the characteristics of MSCs. Further comparative study demonstrated that the characteristics of MC-MSCs were similar to that of bone marrow (BM)-MSCs, including morphology, phenotype, proliferation potential, multilineage differentiation capacity and hematopoiesis-supportive function. The colony-forming unit-fibroblast frequency of the MCs was equivalent to approximately 20-fold of that of the BM. In addition, a BM transplantation experiment demonstrated that MC-MSCs were derived from the peripheral blood. In conclusion, we successfully establish an efficient method to harvest MSCs, and together with the distinct advantages of this method, such as accessibility and possibility for autologous cell therapy, we conclude that our efficient method may be a promising alternative for clinical application.