Analysis of full and empty ratio of EV71 virus by using capillary zone electrophoresis.

Hand, foot, and mouth disease is a serious public health problem, and the main pathogen is enterovirus 71 (EV71). Its capsid assembly mechanism including capsid protein processing has been widely studied. Full and empty capsids have different immunological efficacy. Therefore, tracking full/empty capsid ratio throughout the EV71 production process is important to ensure consistent product quality and proper dosing response. The analysis of full/empty capsid ratio of intact virus has been widely reported as well. A variety of techniques have been employed to evaluate the full/empty capsid ratios. However, there has not been a rapid, reproducible, and robust assay to determine the full/empty capsid ratios of final and in-process products. In this study, a novel assay based on capillary zone electrophoresis was established. The separation of full and empty species could be achieved within 10 min and the ratio of peak areas was used to calculate the full/empty capsid ratio directly. The results showed good reproducibility and linearity for the determination of full/empty capsid ratios.

Pangenome obtained by long-read sequencing of 11 genomes reveal hidden functional structural variants in pigs.

Long-read sequencing (LRS) facilitates both the genome assembly and the discovery of structural variants (SVs). Here, we built a graph-based pig pangenome by incorporating 11 LRS genomes with an average of 94.01% BUSCO completeness score, revealing 206-Mb novel sequences. We discovered 183,352 nonredundant SVs (63% novel), representing 12.12% of the reference genome. By genotyping SVs in an additional 196 short-read sequencing samples, we identified thousands of population stratified SVs. Particularly, we detected 7,568 Tibetan specific SVs, some of which demonstrate significant population differentiation between Tibetan and low-altitude pigs, which might be associated with the high-altitude hypoxia adaptation in Tibetan pigs. Further integrating functional genomic data, the most promising candidate genes within the SVs that might contribute to the high-altitude hypoxia adaptation were discovered. Overall, our study generates a benchmark pangenome resource for illustrating the important roles of SVs in adaptive evolution, domestication, and genetic improvement of agronomic traits in pigs.

A butterfly shaped Eu4(OH)2 cluster-based luminescent metal-organic framework with Lewis basic triazole sites demonstrating turn off sensing in the presence of organic amines.

The effective detection of organic amines is particularly important for environmental protection and human health. Herein, according to hard and soft acid base theory, a novel three-dimensional (3D) butterfly shaped Eu4(OH)2 cluster-based metal-organic framework with Lewis basic triazole sites, {[Eu4(taip)4(ox)(OH)2(H2O)4]·3H2O}n (1) (H2taip = 5-(1,2,4-triazol-1-yl) isophthalic acid, H2ox = oxalic acid), was successfully synthesized under solvothermal conditions, and was characterized by single crystal X-ray diffraction, powder X-ray diffraction, elemental analysis, infrared spectroscopy and thermogravimetric analysis. Structural analysis reveals that compound 1 is a 3D net constructed from butterfly shaped Eu4(OH)2 clusters and contains isosceles triangular channels with dimensions of 8.84 × 8.84 × 8.63 Å3, which shows an unprecedented 8-connected topology with a Schläfli symbol {36·418·53·6}. Fluorescence experiments of compound 1 show sensitive luminescence quenching responses to organic amines such as diethylamine (DEA), trimethylamine (TMA), triethylamine (TEA), ethylenediamine (EDA) and aniline, and the quenching constants (KSV) decrease in the following order: EDA > DEA > TMA > TEA > aniline. The fluorescence quenching responses may be attributed to the energy gap between the LUMO energy values of H2taip and organic amines, which hinders the transfer of excited state energy to the emission state of Eu3+ and results in luminescence quenching. The fluorescence lifetimes of compound 1 in ethanol and organic anilines indicate that the fluorescence recognition process of organic amines was static.

Inhibitory effects of aloperine on voltage-gated Na+ channels in rat ventricular myocytes.

Aloperine (ALO), a quinolizidine alkaloid extracted from Sophora alopecuroides L., modulates hypertension, ventricular remodeling, and myocardial ischemia. However, few studies have evaluated the effects of ALO on other cardiovascular parameters. Accordingly, in this study, we used a rat model of aconitine-induced ventricular arrhythmia to assess the effects of ALO. Notably, ALO pretreatment delayed the onset of ventricular premature and ventricular tachycardia and reduced the incidence of fatal ventricular fibrillation. Moreover, whole-cell patch-clamp assays in rats' ventricular myocyte showed that ALO (3, 10, and 30 µM) significantly reduced the peak sodium current density of voltage-gated Na+ channel currents (INa) in a concentration-dependent manner. The gating kinetics characteristics showed that the steady-state activation and recovery curve were shifted in positive direction along the voltage axis, respectively, and the steady-state inactivation curve was shifted in negative direction along the voltage axis, i.e., which was similar to the inhibitory effects of amiodarone. These results indicated that ALO had anti-arrhythmic effects, partly attributed to INa inhibition. ALO may act as a class I sodium channel anti-arrhythmia agent.

Highly Efficient and Selective Dissolution Separation of Fission Products by an Ionic Liquid [Hbet][Tf2N]: A New Approach to Spent Nuclear Fuel Recycling.

Here, we propose the use of carboxyl-functionalized ionic liquid, [Hbet][Tf2N], to separate the fission products from spent nuclear fuels. This innovative method allows the selective dissolution of neutron poisons, lanthanides oxide, as well as some fission products with high yield, leaving most of the UO2 matrix and minor actinides behind in the spent nuclear fuel and accomplishing the actinides recovery as a group. Water-saturated [Hbet][Tf2N] can dissolve lanthanides oxide from simulated spent nuclear fuel with a dissolution ratio of 100% at 40 °C. However, the dissolution of uranium is almost negligible (<1%) under the same conditions. This big difference in dissolution provides a novel separation approach to spent nuclear fuel recycling and may open new perspectives for spent nuclear fuel reprocessing. The recovery of Nd and U from metal-loaded ionic liquids and the recyclability of the ionic liquid [Hbet][Tf2N] have also been investigated. Furthermore, a U/ x value related to the lattice energy U of metal compound M xO y is used to elaborate the solubility. This work represents the first case for efficient fission products removal by selective dissolution, avoiding the complete dissolution of spent nuclear fuel, the producing of the large high-level radioactive waste, and reducing environmental hazards.

An R2R3-MYB gene, LeAN2, positively regulated the thermo-tolerance in transgenic tomato.

LeAN2 is an anthocyanin-associated R2R3-MYB transcription factor, but little is known about its function in imparting thermo-tolerance to higher plants. To examine the function of LeAN2 in the regulation of heat stress in tomato, LeAN2 was isolated and transgenic tomato plants were obtained. Overexpression of LeAN2 under the control of the CaMV35S promoter in tomato induced the up-regulation of several structural genes in the anthocyanin biosynthetic pathway as well as anthocyanin accumulation in transgenic tomato plants. Transgenic tomato plants showed enhanced tolerance to heat stress by maintaining higher fresh weight (FW), net photosynthetic rate (Pn) and maximal photochemical efficiency of photosystem II (PSII) (Fv/Fm) compared with wild-type (WT) plants. Furthermore, transgenic plants showed higher non-enzymatic antioxidant activity, lower levels of reactive oxygen species (ROS), and higher contents of D1 protein than that in WT plants under heat stress. These results indicate that LeAN2 had an important function in heat stress resistance.

A novel tomato MYC-type ICE1-like transcription factor, SlICE1a, confers cold, osmotic and salt tolerance in transgenic tobacco.

ICE1 (inducer of CBF expression 1), a MYC-type bHLH transcription factor, is an important activator of CBF3/DREB1A for regulating cold signaling and stress tolerance. In this study, we isolated the novel ICE1-like gene SlICE1a from tomato which contains the conserved bHLH domain, an S-rich motif, and ACT-domain. It is localized in the nucleus and harbors transcription-activating activity in the N-terminal. In addition, the SlICE1a transcript is slightly upregulated by cold stress, salt stress, and osmotic stress. SlICE1a overexpression in tobacco enhances the induction of CBF/DREB and their target genes, consequently increasing the levels of proline, soluble sugars, and late embryogenesis abundant (LEA) proteins, and enhancing tolerance to cold stress, osmotic stress, and salt stress. SlICE1a functions in abiotic stress responses by regulating the expression of stress-tolerant genes, and is thus beneficial for crop improvement.

Overexpression of tomato enhancer of SOS3-1 (LeENH1) in tobacco enhanced salinity tolerance by excluding Na+ from the cytosol.

The salt overly sensitive pathway has an important function in plant salinity tolerance. The enhancer of SOS3-1 (ENH1) participates in a new salinity stress pathway with SOS2 but without SOS3. To investigate the physiological effects and functional mechanism of ENH1 under salt stress, ENH1 was isolated from tomato and overexpressed in tobacco. Under salt stress, the sprouting percentage, fresh weight, and dry weight of transgenic plants were higher than those of wild-type (WT) plants. Under salt stress, the chlorophyll content, net photosynthetic rate, and maximal photochemical efficiency of PSII in transgenic plants decreased more slowly than those in WT plants. The overexpression of LeENH1 in tobacco excluded Na(+) from the cytosol and retained high K(+) levels in the cytosol to reestablish ion homeostasis. Higher thylakoid-bound ascorbate peroxidase activity and lower reactive oxygen species levels were found in transgenic plants under salt stress.

[Clinical studies of dynamic changes on the renal injury indicators of acute paraquat poisoning].

OBJECTIVE: To investigate the clinical significance of dynamic changes of blood urea nitrogen (BUN), serum creatinine (Cr), serum cystatin (Cys C) and urinary protein on renal injury with paraquat poisoning. METHODS: According to the clinical manifestation and curative effect, the clinical information was analyzed retrospectively in 35 cases of acute paraquat poisoning, survival after eight weeks as the standard. Poisoning patients were taken a fasting blood 5 ml and the middle of urinary on the 1st day, 3rd day, 7th day, 14th day, 21st day and 8 weeks after the poisoning. Then the levels of serum BUN, Cr, Cys C and urinary protein were detected by automatic biochemistry analyzer. 30 cases healthy subjects were randomly selected as normal control group, and discharged kidney disease and other diseases of urinary system history. The same laboratory subjects have been done. RESULTS: The level of serum Bun, Cr, Cys C of survival group increased significantly compared with control group within 21 days (P < 0.05). The level of serum BUN, Cr Cys C decreased on the 14th day. The decreased level of serum Cys C was lower than that of serum BUN and Cr. The renal function of 29 cases among 35 cases survival patients recovered on 21st day. The renal function of 31 cases among 35 cases survival patients recovered 8 weeks late. The positive rate of urinary protein of survival patients was high in the early intoxication (76.9%). CONCLUSION: Serum Cys C is sensitive indicator to reveal the kidney injury on paraquat poisoning patients and have higher value of clinical applications in the diagnosis of the kidney injury of paraquat poisoning, which sensitivity is higher than serum BUN and Cr. The kidney injury caused by paraquat poisoning is reversible.

Morphology, distribution, and abundance of antennal sensilla of male and female macropterous and brachypterous small brown planthopper, Laodelphax striatellus (Fallén) (Hemiptera: Delphacidae).

The antennal sensilla of both genders of macropterous and brachypterous adults of the small brown planthopper, Laodelphax striatellus (Fallén) (Hemiptera: Delphacidae) were examined using light and scanning electron microscopy. Scanning electron microscopy revealed seven types of antennal sensilla in adult L. striatellus which were not evenly distributed on all antennal segments. Sensilla chaetica, a sensillum campaniformium and a Böhm bristle were found on the scape. Sensilla chaetica, sensilla trichodea, sensilla placodea which always present as plaque organs, sensilla basiconica and a sensillum campaniformium were present on the pedicel. Three sensilla basiconica and one sensillum coeloconicum containing two sensory pegs were located on the swollen sensory region of the basal flagellum. Pores observed on the surface of s. trichodea and s. placodea suggest these organs probably play a role in olfaction, whereas the aporous s. chaetica with flexible sockets probably function as mechanoreceptors. The aporous s. basiconica with inflexible sockets are probable to be thermo-hygroreceptors while the Böhm bristle and s. campaniformia may act as antennal proprioceptors. The function of s. coeloconicum remains uncertain. The numerical dominance of antennal olfactory receptors suggests olfaction is an important function of the antenna in L. striatellus. Although a small degree of sexual/wing dimorphism was observed in the numbers of sensilla and in the length and width of antennae and antennal segments, the basic shape and structure of the antennae and antennal sensilla did not differ between the gender or wing form in L. striatellus.

[Function of a novel brain-specific gene LRRC4].

OBJECTIVE: To study the suppressive effect of LRRC4 gene on human glioma U251 cells and further investigate its biological functions. METHODS: H&E, DNA and AgNORs stainings were performed on LRRC4-transfected U251 cells, mock-transfected U251 cells and non-transfected U251 cells, respectively. Quantitative analysis including cell morphometry, DNA content, DNA ploidy, silver stained argyrophilic nucleolar organizer regions (AgNORs) were investigated by image analysis. Flow cytometry was employed to determine the difference of cell cycle distribution and MTT staining was used to elucidate the activity of the LRRC4-transfected U251 cells. RESULTS: The morphological cell parameters such as area, perimeter and diameter, DNA content, chromosomal aneupoloidy, mean area of AgNORs particles and mean nucleus area of the LRRC4-transfected U251 cells were remarkably decreased compared to those of the mock-transfected and non-transfected U251 cells (P < 0.05, P < 0.01). Meanwhile, significant accumulation of cells in G(0)/G(1) phase but decrease of cells in S and G(2)/M phase, was observed in transfected U251 cells compared to those of the mock-transfected and non-transfected U251 cells (P < 0.05, P < 0.01). MTT staining showed that proliferation activity of both the mock- and non-trasfected U251 cells was significantly higher than that of the U251 cells transfected with LRRC4 gene (P < 0.01). CONCLUSION: LRRC4 gene might be involved in tumor suppression by restraining DNA synthesis and the nucleoli organizer regions-associated proteins, keeping the cell cycles in phase G(0)/G(1) and reducing proliferation activity of the glioma cells. Morphometry combined with other techniques such as flow cytometry and MTT staining can well elucidate the biological function of novel genes.

Study of a novel brain relatively specific gene LRRC4 involved in glioma tumorigenesis suppression using the Tet-on system.

LRRC4 is a novel relatively specific gene, which displays significant down-regulation in primary brain tumor biopsies and has the potential to suppress brain tumor growth. In this study, we investigated the growth inhibitory effect of LRRC4 on tumorigencity in vivo and on cell proliferation in vitro by a tetracycline-inducible expression system. Results showed that LRRC4 significantly reduced the growth and malignant grade of xenografts arising from glioblastoma U251MG cells. Cell proliferation was markedly inhibited after U251MG Tet-on-LRRC4 cell induction with doxycycline. Flow cytometry and Western blot analysis demonstrated that LRRC4 mediated a delay of the cell cycle in late G1, possibly through up-regulating the expressions of p21Waf1/cip1 and p27Kip1 and down-regulating the expressions of cyclin-dependent kinase 2, retinoblastoma protein and epidermal growth factor receptors. Together, these findings provide clues to the function of LRRC4 as a negative regulator of cell growth and underscore a link between the above-mentioned cyclins, cyclin-associated molecules and tumorigenicity.

Expression of tumor related gene NAG6 in gastric cancer and restriction fragment length polymorphism analysis.

AIM: NAG6 gene is a novel tumor related gene identified recently. This study was designed to examine the expression of this gene in gastric cancer and corresponding normal tissues, and to investigate its role in the occurrence and development of gastric cancer, also to study if the genetic structure of NAG6 was altered in gastric cancer. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR), Northern blot analysis and dot hybridization were used to compare the expression level of NAG6 gene in 42 cases of gastric cancer tissues with their corresponding normal tissues of the same patients respectively. In addition, restriction fragment length polymorphism (RFLP) analysis was adopted to study if the genetic structure of NAG6 was altered in gastric carcinomas. RESULTS: The expression of NAG6 in 57.1% gastric cancer tissues (25/42) was absent by RT-PCR analysis. The down-regulation rate of NAG6 in gastric cancer tissues was significantly higher than that in corresponding normal tissues (P<0.01). However no correlation between the down-regulation of NAG6 and lymph-node and/or distance metastasis was found in this study (P>0.05). Dot hybridization confirmed the results of RT-PCR. Furthermore, the results of EcoRI RFLP analysis of NAG6 gene demonstrated that 3 of 7 cases of gastric cancer showed loss of 5 kb fragment in comparison with their corresponding normal tissues. CONCLUSION: NAG6 gene is significantly down regulated in gastric cancer. The loss of genetic materials may be the cause of down-regulation of NAG6 expression. This seems to suggest that NAG6 may represent a candidate of putative tumor suppressor gene at 7q31-32 loci associated with gastric carcinoma. The down-regulation of this gene may play a role in occurrence and development of this disease, however it may not be associated with lymph node and/or distance metastasis.

A susceptibility locus at chromosome 3p21 linked to familial nasopharyngeal carcinoma.

Nasopharyngeal carcinoma (NPC) poses one of the serious health problems in southern Chinese, with an incidence rate ranging from 15 to 50/100,000. Chromosome translocation t(1;3) and frequent loss of heterogeneity on short arms of chromosome 3 and 9 have been reported to be associated with NPC, and a genome-wide scan identified an NPC susceptibility locus on chromosome 4p15.1-q12 recently. In our study, we collected samples from 18 families at high risk of NPC from the Hunan province in southern China, genotyped with a panel of polymorphic markers on short arms of chromosomes 3, 9, and 4p15.1-q12. A locus on 3p21 was identified to link to NPC with a maximum logarithm of odds for linkage score of 4.18. Fine mapping located the locus to a 13.6-cM region on 3p21.31-21.2, where a tumor suppressor gene cluster resided. Our findings identified a novel locus for NPC and provided a map location for susceptibility genes candidates. In contrast to a recent study, no significant evidence for NPC linkage to chromosomes 4 and 9 was observed.

Expression of LRRC4 has the potential to decrease the growth rate and tumorigenesis of glioblastoma cell line U251.

BACKGROUND & OBJECTIVE: LRRC4 is a novel gene that the author has identified recently, which displayed significant downregulation in primary brain tumor biopsies. This study was designed to investigate if LRRC4 has the potential of suppressing brain tumor growth. METHODS: The full-length coding region of LRRC4 gene was subcloned into the expression vector pcDNA3.1, the recombinant was introduced into the glioblastoma cell line U251 by liposome transfection, and the U251 cells stably expressing LRRC4 gene were established by G418 selection. Furthermore, cell proliferation assay, soft agar assay, tumorigenesis assay were taken to examine the effect of LRRC4 expression on cell growth and tumor formation. RESULTS: U251 cells stably expressing full-length coding region of LRRC4 were established by lipofection-mediated transfection and selected for further study. Compared with the nontransfected and vector-transfected cells, the cells transfected with LRRC4 cDNA exhibited a significant increase of expression of LRRC4 mRNA by Northern blot analysis. Further, when cell proliferation was followed over several days, the cells expressing the transfected LRRC4 cDNA grew more slowly than nontransfected cells. Consistently, the cells transfected with LRRC4 exhibited markedly lower colony formation rate. These clones were injected into athymic nude mice who was killed after 40 days and the tumor sizes were evaluated. Tumor volume in mice was significantly smaller in the group of cells stably transfected with LRRC4 cDNA than in the control. CONCLUSION: LRRC4 gene may be transfected into the human glioblastoma cell line U251. The expression of LRRC4 in U251 cells may have the potential to suppress tumor cell growth and the tumorigenesis of U251 cell transplanted in nude mice.