Macrophage-hitchhiked arsenic/AB bionic preparations for liver cancer.

Macrophage-hitchhiked arsenic/AB bionic preparations were developed to improve the therapeutic effect on liver cancer by means of the tumor-targeting ability of macrophages in vivo. In vitro and in vivo cellular uptake assays demonstrated that arsenic/AB, with negatively charged particles of around 100-200 nm size, could hitchhike to macrophages. Dissolution experiments of arsenic/AB showed that arsenic/AB could delay the release of arsenic and ensure the safety of macrophages during its transport. Histological examination confirmed the safety of the preparations for major organs. In vivo distribution experiment showed that the arsenic/AB bionic preparations could rapidly accumulate in tumors, and in vivo treatment experiment showed a significant tumor inhibition of arsenic/AB. The therapeutic mechanism of liver cancer might be that the arsenic/AB bionic preparations could inhibit tumor growth by reducing inflammatory response and inhibiting CSF1 secretion to block CSF1R activation to induce more differentiation of tumor-associated macrophages (TAMs) towards the anti-tumor M1 phenotype. Therefore, we concluded that the arsenic/AB bionic preparations could improve the distribution of arsenic in vivo by hitchhiking on macrophages as well as make it have tumor targeting and deep penetration abilities, thus increasing the therapeutic effect of arsenic on liver cancer with reduced side effects.

Traumatic dislocation of the iris into the vitreous cavity with intact lens: a case report.

BACKGROUND: Traumatic aniridia occurs when the iris is extruded from the eye and is often accompanied by lens injuries. However, traumatic aniridia due to dislocation of the iris into the vitreous cavity without lens damage has never been reported. CASE PRESENTATION: A 30-year-old man presented with visual loss and pain for 6 h after a thin wire injured his right eyeball. Ophthalmologic examinations manifested a 2 mm full-thickness corneal laceration and total hyphema. An intact clear lens, healthy attached retina, and almost complete iris tissue in the vitreous cavity were found after resolution of hyphema the next day. Further examination revealed that the defect in the zonule below the corneal wound was the path for the iris to enter the vitreous cavity. The patient opted for nonsurgical treatment until pigment granules and opacity were observed in the vitreous cavity after 50 days. Vitrectomy was performed to remove the dislocated iris. CONCLUSIONS: The presentation of this unique case indicates that the torn iris was displaced to the vitreous cavity with an intact lens and missing local zonula instead of out the corneal laceration after a penetrating injury. The type of injury, mechanism, and force on the spot may contribute to the occurrence of this rare condition. Instead of artificial irises, tinted glasses were more appropriate treatment option for this patient. Peripheral retinal examination was essential in the management of this case. In such cases, the iris in the vitreous cavity should be resected to prevent complications.

Niche Filling Dynamics of Ragweed (Ambrosia artemisiifolia L.) during Global Invasion.

Determining whether the climatic ecological niche of an invasive alien plant is similar to that of the niche occupied by its native population (ecological niche conservatism) is essential for predicting the plant invasion process. Ragweed (Ambrosia artemisiifolia L.) usually poses serious threats to human health, agriculture, and ecosystems within its newly occupied range. We calculated the overlap, stability, unfilling, and expansion of ragweed's climatic ecological niche using principal component analysis and performed ecological niche hypothesis testing. The current and potential distribution of A. artemisiifolia was mapped by ecological niche models to identify areas in China with the highest potential risk of A. artemisiifolia invasion. The high ecological niche stability indicates that A. artemisiifolia is ecologically conservative during the invasion. Ecological niche expansion (expansion = 0.407) occurred only in South America. In addition, the difference between the climatic and native niches of the invasive populations is mainly the result of unpopulated niches. The ecological niche model suggests that southwest China, which has not been invaded by A. artemisiifolia, faces an elevated risk of invasion. Although A. artemisiifolia occupies a climatic niche distinct from native populations, the climatic niche of the invasive population is only a subset of the native niche. The difference in climatic conditions is the main factor leading to the ecological niche expansion of A. artemisiifolia during the invasion. Additionally, human activities play a substantial role in the expansion of A. artemisiifolia. Alterations in the A. artemisiifolia niche would help explain why this species is so invasive in China.

Novel mutations in the BEST1 gene cause distinct retinopathies in two Chinese families.

AIM: To describe the clinical heterogeneity of patients with novel mutations in BEST1. METHODS: All the members in the two Chinese families underwent detailed clinical evaluations including best-corrected visual acuity, slit-lamp examination, applanation tonometry, and dilated fundus examination. Fundus autofluorescence, fundus fluorescein angiography, spectral-domain optical coherence tomography, electrooculography, and electroretinogram were also performed. Genomic DNA was extracted from venous blood for all the participants. The targeted next-generation sequencing of inherited retinal disease-associated genes was conducted to identify the causative mutation. RESULTS: A novel BEST1 missense mutation c.41T>C (p.Leu14Ser) was identified in Family 1. It was co-segregated with the phenotype of best vitelliform macular dystrophy (BVMD) and bioinformatics analysis confirmed it was harmful. Another novel BEST1 frameshift mutation c.345_346insGGCAAGGACG (p.Glu119Glyfs*116) and a novel USH2A missense mutation c.12560G>A, p.Arg4187His were identified in family 2 with retinitis pigmentosa (RP), which might interact and lead to the phenotype of RP. CONCLUSION: Two novel mutations in the BEST1 gene in two unrelated families with distinct phenotypes and BEST1 mutation accompanied with USH2A mutation would result in RP, which could be enormously helpful in understanding the pathogenesis of the inherited retinal disease caused by a BEST1 mutation.

The dispersal-related traits of an invasive plant Galinsoga quadriradiata correlate with elevation during range expansion into mountain ranges.

Detecting shifts in trait values among populations of an invasive plant is important for assessing invasion risks and predicting future spread. Although a growing number of studies suggest that the dispersal propensity of invasive plants increases during range expansion, there has been relatively little attention paid to dispersal patterns along elevational gradients. In this study, we tested the differentiation of dispersal-related traits in an invasive plant, Galinsoga quadriradiata, across populations at different elevations in the Qinling and Bashan Mountains in central China. Seed mass-area ratio (MAR), an important seed dispersal-related trait, of 45 populations from along an elevational gradient was measured, and genetic variation of 23 populations was quantified using inter-simple sequence repeat (ISSR) markers. Individuals from four populations were then planted in a greenhouse to compare their performance under shared conditions. Changing patterns of seed dispersal-related traits and populations genetic diversity along elevation were tested using linear regression. Mass-area ratio of G. quadriradiata increased, while genetic diversity decreased with elevation in the field survey. In the greenhouse, populations of G. quadriradiata sourced from different elevations showed a difference response of MAR. These results suggest that although rapid evolution may contribute to the range expansion of G. quadriradiata in mountain ranges, dispersal-related traits will also likely be affected by phenotypic plasticity. This challenges the common argument that dispersal ability of invasive plants increases along dispersal routes. Furthermore, our results suggest that high-altitude populations would be more effective at seed dispersal once they continue to expand their range downslope on the other side. Our experiment provides novel evidence that the spread of these high-altitude populations may be more likely than previously theorized and that they should thus be cautiously monitored.

Comprehensive circRNA-microRNA-mRNA network analysis revealed the novel regulatory mechanism of Trichosporon asahii infection.

BACKGROUND: Invasive Trichosporon asahii (T. asahii) infection frequently occurs with a high mortality in immunodeficient hosts, but the pathogenesis of T. asahii infection remains elusive. Circular RNAs (circRNAs) are a type of endogenous noncoding RNA that participate in various disease processes. However, the mechanism of circRNAs in T. asahii infection remains completely unknown. METHODS: RNA sequencing (RNA-seq) was performed to analyze the expression profiles of circRNAs, microRNAs (miRNAs), and mRNAs in THP-1 cells infected with T. asahii or uninfected samples. Some of the RNA-seq results were verified by RT-qPCR. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were used to analyze the differentially expressed mRNAs. A circRNA-miRNA-mRNA network was constructed and verified by dual-luciferase reporter assay and overexpression experiments. RESULTS: A total of 46 circRNAs, 412 mRNAs and 47 miRNAs were differentially expressed at 12 h after T. asahii infection. GO and KEGG analyses showed that the differentially expressed mRNAs were primarily linked to the leukocyte migration involved in the inflammatory response, the Toll-like receptor signaling pathway, and the TNF signaling pathway. A competing endogenous RNA (ceRNA) network was constructed with 5 differentially expressed circRNAs, 5 differentially expressed miRNAs and 42 differentially expressed mRNAs. Among them, hsa_circ_0065336 was found to indirectly regulate PTPN11 expression by sponging miR-505-3p. CONCLUSIONS: These data revealed a comprehensive circRNA-associated ceRNA network during T. asahii infection, thus providing new insights into the pathogenesis of the T. asahii-host interactions.

Arbuscular mycorrhizal colonization rate of an exotic plant, Galinsoga quadriradiata, in mountain ranges changes with altitude.

Arbuscular mycorrhizal fungi (AMF) are suggested to be important for invasions by many exotic plants. However, it is not yet known how associations between AMF and invasive plant populations change in mountains ranges and how changed associations affect further expansion of different populations in new habitats. We conducted a field survey to detect AMF colonization rate of the invasive Galinsoga quadriradiata along an elevational gradient ranging from 223 to 1947 masl in the Qinling and Bashan Mountains, China. Additionally, a greenhouse experiment was conducted to compare plant growth performance among five elevational populations. In the field, total plant mass and seed production, as well as root AMF colonization rate, significantly decreased with elevation. When populations were grown in a novel soil environment in the greenhouse, the high-altitude populations achieved higher seed and total mass at lower AMF colonization rate than the low-altitude populations. Moreover, high AMF association was related to high intraspecific competition within low-altitude populations and limited seed production. Our results revealed that the associations between AMF and G. quadriradiata decrease with altitude in mountain ranges, and this may indicate that differentiation of association between AMF and elevational populations occurs during range expansion of G. quadriradiata. The results of the greenhouse experiment suggest that the high-altitude populations are more aggressive than the low-altitude populations in a non-stressful environment.

Controllable Epitaxial Growth of MoSe2 Bilayers with Different Stacking Orders by Reverse-Flow Chemical Vapor Deposition.

The stacking order plays a critical role in the electronic and optical properties of two-dimensional materials. It is however of great challenge to achieve large-size and homogeneous bilayer crystals with precisely controlled stacking orders. Here, we demonstrate an optimized chemical vapor deposition strategy to grow MoSe2 bilayers with controlled AA or AB stacking sequences. Reverse gas flow effectively suppresses the random nucleation centers, leading to uniform growth of the second layer of MoSe2 on the first monolayer. A customized temperature profile selectively activates the growth of the MoSe2 bilayer with different stacking orders: the AA stacking MoSe2 bilayer tends to form at 810 °C, and the AB stacking MoSe2 bilayer prefers to grow at a higher temperature of 860 °C. A series of characterization methods confirm that MoSe2 bilayers with different stacking orders exhibit distinct crystal structures and physical properties. Our results demonstrate a robust and effective route for the controllable synthesis of transition metal dichalcogenide bilayers, which will benefit the development of two-dimensional materials and van der Waals heterostructures.

[Preparation of angiopep-2-modified FITC-labeled neurotoxin nanoparticles and in vitro release characteristics].

In order to prepare angiopep-2 modified fluorescein isothiocyanate-labeled neurotoxin nanoparticles( ANG-NPs/FITCNT),emulsion/solvent evaporation method was used with m PEG-PLA and ANG-PEG-PLA( in proper proportions) as carriers and with FITC-NT as drug. With particle size and encapsulation efficiency as comprehensive indexes,the effects of different ultrasound power and ultrasound time combinations on the process were investigated. The in vitro release characteristics of nanoparticles in PBS buffer at p H 7. 4 and p H 6. 5 were investigated by dialysis method. The results indicated that the optimum process for preparing ANG-NPs/FITC-NT was as follows: ultrasonic power 90 W,ultrasonic time 30 s. In such optimal process,ANG-NPs/FITC-NT were well-shaped under the transmission electron microscope,with an average particle size of( 123. 9±0. 5) nm,Zeta potential of(-10. 5±0. 5) m V,encapsulation efficiency of( 68. 1±0. 4) %,and the drug loading of( 0. 82±0. 01) %. The in vitro drug release profiles of the nanoparticles in PBS buffer at p H 7. 4 and p H 6. 5 were both consistent with Ritger-Peppas equation,ln Q = 0. 508 8 lnt-2. 285 0,r = 0. 961 5( p H 7. 4) and ln Q= 0. 449 9 lnt-1. 855 3,r = 0. 970 3( p H 6. 5),respectively. The experiment results proved that the nanoparticles prepared by emulsion/solvent evaporation method had uniform particle size,high encapsulation efficiency and in vitro sustained release characteristic,which might be a potential carrier for NT intracerebral drug delivery.

Elevated nitrogen allows the weak invasive plant Galinsoga quadriradiata to become more vigorous with respect to inter-specific competition.

Elevated nitrogen associated with global change is believed to promote the invasion of many vigorous exotic plants. However, it is unclear how a weak exotic plant will respond to elevated nitrogen in the future. In this study, the competitive outcome of a weak invasive plant (Galinsoga quadriradiata) and two non-invasive plants was detected. The plants were subjected to 3 types of culture (mixed, monoculture or one-plant), 2 levels of nitrogen (ambient or elevated at a rate of 2 g m-2 yr-1) and 2 levels of light (65% shade or full sunlight). The results showed that elevated nitrogen significantly promoted the growth of both the weak invader and the non-invasive plants in one-plant pots; however, growth promotion was not observed for the non-invasive species in the mixed culture pots. The presence of G. quadriradiata significantly inhibited the growth of the non-invasive plants, and a decreased negative species interaction was detected as a result of elevated nitrogen. Our results suggest that competitive interactions between G. quadriradiata and the non-invasive plants were altered by elevated nitrogen. It provides exceptional evidence that an initially weak invasive plant can become an aggressive invader through elevated nitrogen deposition.

Proteomic profiling of early degenerative retina of RCS rats.

AIM: To identify the underlying cellular and molecular changes in retinitis pigmentosa (RP). METHODS: Label-free quantification-based proteomics analysis, with its advantages of being more economic and consisting of simpler procedures, has been used with increasing frequency in modern biological research. Dystrophic RCS rats, the first laboratory animal model for the study of RP, possess a similar pathological course as human beings with the diseases. Thus, we employed a comparative proteomics analysis approach for in-depth proteome profiling of retinas from dystrophic RCS rats and non-dystrophic congenic controls through Linear Trap Quadrupole - orbitrap MS/MS, to identify the significant differentially expressed proteins (DEPs). Bioinformatics analyses, including Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotation and upstream regulatory analysis, were then performed on these retina proteins. Finally, a Western blotting experiment was carried out to verify the difference in the abundance of transcript factor E2F1. RESULTS: In this study, we identified a total of 2375 protein groups from the retinal protein samples of RCS rats and non-dystrophic congenic controls. Four hundred thirty-four significantly DEPs were selected by Student's t-test. Based on the results of the bioinformatics analysis, we identified mitochondrial dysfunction and transcription factor E2F1 as the key initiation factors in early retinal degenerative process. CONCLUSION: We showed that the mitochondrial dysfunction and the transcription factor E2F1 substantially contribute to the disease etiology of RP. The results provide a new potential therapeutic approach for this retinal degenerative disease.

Ultra-fast pulse propagation in nonlinear graphene/silicon ridge waveguide.

We report the femtosecond laser propagation in a hybrid graphene/silicon ridge waveguide with demonstration of the ultra-large Kerr coefficient of graphene. We also fabricated a slot-like graphene/silicon ridge waveguide which can enhance its effective Kerr coefficient 1.5 times compared with the graphene/silicon ridge waveguide. Both transverse-electric-like (TE-like) mode and transverse-magnetic-like (TM-like) mode are experimentally measured and numerically analyzed. The results show nonlinearity dependence on mode polarization not in graphene/silicon ridge waveguide but in slot-like graphene/silicon ridge waveguide. Great spectral broadening was observed due to self-phase modulation (SPM) after propagation in the hybrid waveguide with length of 2 mm. Power dependence property of the slot-like hybrid waveguide is also measured and numerically analyzed. The results also confirm the effective Kerr coefficient estimation of the hybrid structures. Spectral blue shift of the output pulse was observed in the slot-like graphene/silicon ridge waveguide. One possible explanation is that the blue shift was caused by the ultra-fast free carrier effect with the optical absorption of the doped graphene. This interesting effect can be used for soliton compression in femtosecond region. We also discussed the broadband anomalous dispersion of the Kerr coefficient of graphene.

Mixed Linear Model Approaches of Association Mapping for Complex Traits Based on Omics Variants.

Precise prediction for genetic architecture of complex traits is impeded by the limited understanding on genetic effects of complex traits, especially on gene-by-gene (GxG) and gene-by-environment (GxE) interaction. In the past decades, an explosion of high throughput technologies enables omics studies at multiple levels (such as genomics, transcriptomics, proteomics, and metabolomics). The analyses of large omics data, especially two-loci interaction analysis, are very time intensive. Integrating the diverse omics data and environmental effects in the analyses also remain challenges. We proposed mixed linear model approaches using GPU (Graphic Processing Unit) computation to simultaneously dissect various genetic effects. Analyses can be performed for estimating genetic main effects, GxG epistasis effects, and GxE environment interaction effects on large-scale omics data for complex traits, and for estimating heritability of specific genetic effects. Both mouse data analyses and Monte Carlo simulations demonstrated that genetic effects and environment interaction effects could be unbiasedly estimated with high statistical power by using the proposed approaches.

[Intestinal absorption kinetics of flurbiprofen in rats].

To study the in situ intestinal absorption kinetics of flrubiprofen in rats, the absorption of flurbiprofen in small intestine (duodenum, jejunum and ileum) and colon of rats was investigated using in situ single-pass perfusion method and the drug content was measured by HPLC. The effects of drug concentration on the intestinal absorption were investigated. The K(a) and P(app) values of flurbiprofen in the small intestine and colon had no significant difference (P > 0.05). Drug concentration (4.0, 10.0 and 16.0 mg x L(-1)) had no significant influence on the K(a) values (P > 0.05). However, when concentration was 4.0 mg x L(-1) and 10.0 mg x L(-1), significant effect on the P(app) values (P < 0.05) was found, but significant effect on the P(app) values was not shown between 10.0 mg x L(-1) and 16.0 mg x L(-1) (P > 0.05). The K(a) and P(app) values of flurbiprofen on the perfusion flow rate had significant difference (P < 0.05). Flurbiprofen could be absorbed at all segments of the intestine in rats and had no special absorption window. The absorption of flurbiprofen complies with the facilitated diffusion in the general intestinal segments, and accompany with the cytopsistransport mechanism probably. The perfusion flow rate had significant effect on the K(a) and P(app).

Design of ultrathin plasmonic quarter-wave plate based on period coupling.

Based on an analysis of the surface admittance of a plasmonic film with a substrate, we propose an ultrathin quarter-wave plate consisting of a periodic plane array of symmetrical L-shaped plasmonic antennas. The period, which determines the coupling among L-shaped antennas, is an important parameter for optimizing the performance of the structure. Numerical simulation results show that an Au quarter-wave plate designed in this Letter can efficiently convert a linearly polarized light at normal incidence into circularly polarized light, whose ellipticity is 0.994 at an operating wavelength of 1550 nm. The thickness is only 30 nm, which is nearly 1/50 of the wavelength of incident light.

Improving the room-temperature confinement of light by miniaturizing mode sizes into a deep subwavelength scale using dielectric spheres in metal cavities.

The confinement of light within nanometer-scale regions may result in the significant enhancement of light-matter interactions. However, light confinement to nanometers is hindered by the diffraction limit of a dielectric material. For a dielectric cavity, if the material loss is negligible, reducing the cavity size usually causes a significantly increase in radiation loss. Surface plasmons show great promise for potential subwavelength light confinement. However, in most circumstances, light confinement by dissipative metallic materials can cause ohmic losses at optical frequencies. In such cases, the realization of light confinement with deep subwavelength mode sizes results in great losses and thus has low quality factors. In the present study, a three-dimensional light confinement with deep subwavelength mode sizes is achieved using dielectric spheres in metal cavities. Contrary to other mechanisms for subwavelength light confinement that are based on the use of dielectric or metal cavities, the nanometer-scale regions ensure that most of the light energy is confined away from the metal-dielectric interfaces, thereby decreasing light absorption in the metal cavity. In turn, the metal cavity decreases the radiation loss of light. Thus, high quality factors ranging from 2×10(2) to 6×10(2) can be obtained at room temperature. An effective electrical mode volume ranging from 7×10(-5)λ(0)(3) to 2×10(-4)λ(0)(3) (where λ(0) is the resonant wavelength in a vacuum) can be achieved. Therefore, this method of three-dimensional light confinement with deep subwavelength mode sizes using dielectric spheres in metal cavities may have potential applications in the design of nanolasers, nanophoton detectors, nonlinear optical switches, and so on.

Wave propagation in deep-subwavelength mode waveguides.

This Letter proposes a dielectric waveguide with deep-subwavelength mode sizes. Results of both frequency domain and time domain analysis show that the effective mode area is below λ(0)(2)/400 and can even reach λ(0)(2)/1000 (λ(0) is the wavelength in vacuum). The effective electrical mode area can be comparable to that of a hybrid plasmonic subwavelength confinement waveguide, with reduced optical absorption. In contrast to slot waveguides, which guide light in low-index materials, the proposed structure guides light in high-index materials. Results obtained in this Letter show that the losses are sensitive to the surface roughness on the tens of nanometers scale. The structure can be used to design ring resonators with a quality factor comparable to that of a diffraction-limited dielectric ring resonator with the same standing wave numbers. The property can be applied in nonlinear effect enhancement or laser design with ultralow threshold.

A new mapping method for quantitative trait loci of silkworm.

BACKGROUND: Silkworm is the basis of sericultural industry and the model organism in insect genetics study. Mapping quantitative trait loci (QTLs) underlying economically important traits of silkworm is of high significance for promoting the silkworm molecular breeding and advancing our knowledge on genetic architecture of the Lepidoptera. Yet, the currently used mapping methods are not well suitable for silkworm, because of ignoring the recombination difference in meiosis between two sexes. RESULTS: A mixed linear model including QTL main effects, epistatic effects, and QTL × sex interaction effects was proposed for mapping QTLs in an F2 population of silkworm. The number and positions of QTLs were determined by F-test and model selection. The Markov chain Monte Carlo (MCMC) algorithm was employed to estimate and test genetic effects of QTLs and QTL × sex interaction effects. The effectiveness of the model and statistical method was validated by a series of simulations. The results indicate that when markers are distributed sparsely on chromosomes, our method will substantially improve estimation accuracy as compared to the normal chiasmate F2 model. We also found that a sample size of hundreds was sufficiently large to unbiasedly estimate all the four types of epistases (i.e., additive-additive, additive-dominance, dominance-additive, and dominance-dominance) when the paired QTLs reside on different chromosomes in silkworm. CONCLUSION: The proposed method could accurately estimate not only the additive, dominance and digenic epistatic effects but also their interaction effects with sex, correcting the potential bias and precision loss in the current QTL mapping practice of silkworm and thus representing an important addition to the arsenal of QTL mapping tools.

Bidirectional mediation of TiO2 nanowires field effect transistor by dipole moment from purple membrane.

Bacteriorhodopsin-embedded purple membrane (bR-PM) is one of the most promising biomaterials for various bioelectronics applications. In this work, we demonstrate that a dipole bio-originated from bR-PM can bidirectionally mediate the performance of a bottom-contact TiO(2) nanowire field effect transistor (FET) for performance improvement. When negative gate voltage is applied, both transfer and output characteristics of the TiO(2) nanowire FET are enhanced by the bR-PM modification, resulting in a hole mobility increased by a factor of 2. The effect of the number of the deposited bR-PM layers on the normalized DeltaI(D) of the FET suggests that the additional electric field generated by the dipole moment natively existing in bR-PM actually boosts the performance of the TiO(2) nanowires FET.

Resonance amplification of left-handed transmission at optical frequencies by stimulated emission of radiation in active metamaterials.

We demonstrate that left-handed resonance transmission from metallic metamaterial, composed of periodically arranged double rings, can be extended to visible spectrum by introducing an active medium layer as the substrate. The severe ohmic loss inside metals at optical frequencies is compensated by stimulated emission of radiation in this active system. Due to the resonance amplification mechanism of recently proposed lasing spaser, the left-handed transmission band can be restored up to 610 nm wavelength, in dependence on the gain coefficient of the active layer. Additionally, threshold gains for different scaling levels of the double-ring unit are investigated to evaluate the gain requirement of left-handed transmission restoration at different frequency ranges.