A simple and efficient CRISPR/Cas9 system permits ultra-multiplex genome editing in plants

The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sg RNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sg RNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering.

Identification of Mixtures of Two Types of Body Fluids Using the Multiplex Methylation System and Random Forest Models

Objective Body fluid mixtures are complex biological samples that frequently occur in crime scenes,and can provide important clues for criminal case analysis.DNA methylation assay has been applied in the identification of human body fluids,and has exhibited excellent performance in predicting single-source body fluids.The present study aims to develop a methylation SNaPshot multiplex system for body fluid identification,and accurately predict the mixture samples.In addition,the value of DNA methylation in the prediction of body fluid mixtures was further explored.Methods In the present study,420 samples of body fluid mixtures and 250 samples of single body fluids were tested using an optimized multiplex methylation system.Each kind of body fluid sample presented the specific methylation profiles of the 10 markers.Results Significant differences in methylation levels were observed between the mixtures and single body fluids.For all kinds of mixtures,the Spearman’s correlation analysis revealed a significantly strong correlation between the methylation levels and component proportions(1:20,1:10,1:5,1:1,5:1,10:1 and 20:1).Two random forest classification models were trained for the prediction of mixture types and the prediction of the mixture proportion of 2 components,based on the methylation levels of 10 markers.For the mixture prediction,Model-1 presented outstanding prediction accuracy,which reached up to 99.3%in 427 training samples,and had a remarkable accuracy of 100%in 243 independent test samples.For the mixture proportion prediction,Model-2 demonstrated an excellent accuracy of 98.8%in 252 training samples,and 98.2%in 168 independent test samples.The total prediction accuracy reached 99.3%for body fluid mixtures and 98.6%for the mixture proportions.Conclusion These results indicate the excellent capability and powerful value of the multiplex methylation system in the identification of forensic body fluid mixtures.

A High Spectral Efficient Frequency-Domain Channel-Estimation Method for the Polarization-Division-Multiplexed CO-OFDM-OQAM System

Contrary to the other multi-carrier modulation systems, the coherent optical orthogonal frequency division multiplexing communication system with an offset quadrature amplitude modulation (CO-OFDM-OQAM) possesses inherent imaginary interference (IMI). This has an important impact on the channel estimation process. Currently, a variety of frequency-domain channel estimation methods have been proposed. However, there are various problems that still exist. For instance, in order to reduce the influence of IMI, it is necessary to insert more guard intervals between the training sequence and the payload, leading to the occupation of excessive spectrum resources. In order to address this problem, this work designs a high spectral efficient frequency-domain channel estimation method for the polarization-division-multiplexing CO-OFDM-OQAM systems. First, the working principle of the proposed method is described in detail. Then, its spectral efficiency, power peak-to-average ratio, and channel estimation performance are studied based on simulations. The simulation results show that the proposed method improves the spectral efficiency without worsening the power peak-to-average ratio. The channel estimation capability of this method is verified in three scenarios of long-distance transmissions, including back-to-back, 100 km, and 200 km transmissions. .

Multi-dimensional multiplexing optical secret sharing framework with cascaded liquid crystal holograms

Secret sharing is a promising technology for information encryption by splitting the secret information into different shares.However,the traditional scheme suffers from information leakage in decryption process since the amount of available information channels is limited.Herein,we propose and demonstrate an optical secret sharing framework based on the multi-dimensional multiplexing liquid crystal(LC)holograms.The LC holograms are used as spatially separated shares to carry secret images.The polarization of the incident light and the distance between different shares are served as secret keys,which can significantly improve the information security and capacity.Besides,the decryption condition is also restricted by the applied external voltage due to the variant diffraction efficiency,which further increases the information security.In implementation,an artificial neural network(ANN)model is developed to carefully design the phase distribution of each LC hologram.With the advantage of high security,high capacity and simple configuration,our optical secret sharing framework has great potentials in optical encryption and dynamic holographic display.

Semi-implantable device based on multiplexed microfilament electrode cluster for continuous monitoring of physiological ions

Modern medicine is increasingly interested in advanced sensors to detect and analyze biochemical indicators.Ion sensors based on potentiometric methods are a promising platform for monitoring physiological ions in biological subjects.Current semi-implantable devices are mainly based on single-parameter detection.Miniaturized semi-implantable electrodes for multiparameter sensing have more restrictions on the electrode size due to biocompatibility considerations,but reducing the electrode surface area could potentially limit electrode sensitivity.This study developed a semi-implantable device system comprising a multiplexed microfilament electrode cluster(MMEC)and a printed circuit board for real-time monitoring of intra-tissue K^(+),Ca^(2+),and Na^(+)concentrations.The electrode surface area was less important for the potentiometric sensing mechanism,suggesting the feasibility of using a tiny fiber-like electrode for potentiometric sensing.The MMEC device exhibited a broad linear response(K^(+):2–32 mmol/L;Ca^(2+):0.5–4 mmol/L;Na^(+):10–160 mmol/L),high sensitivity(about 20–45 mV/decade),temporal stability(>2weeks),and good selectivity(>80%)for the above ions.In vitro detection and in vivo subcutaneous and brain experiment results showed that the MMEC system exhibits good multi-ion monitoring performance in several complex environments.This work provides a platform for the continuous real-time monitoring of ion fluctuations in different situations and has implications for developing smart sensors to monitor human health.

Turbo Message Passing Based Burst Interference Cancellation for Data Detection in Massive MIMO-OFDM Systems

This paper investigates the fundamental data detection problem with burst interference in massive multiple-input multiple-output orthogonal frequency division multiplexing(MIMO-OFDM) systems. In particular, burst interference may occur only on data symbols but not on pilot symbols, which means that interference information cannot be premeasured. To cancel the burst interference, we first revisit the uplink multi-user system and develop a matrixform system model, where the covariance pattern and the low-rank property of the interference matrix is discussed. Then, we propose a turbo message passing based burst interference cancellation(TMP-BIC) algorithm to solve the data detection problem, where the constellation information of target data is fully exploited to refine its estimate. Furthermore, in the TMP-BIC algorithm, we design one module to cope with the interference matrix by exploiting its lowrank property. Numerical results demonstrate that the proposed algorithm can effectively mitigate the adverse effects of burst interference and approach the interference-free bound.

Polar codes based OFDM-PLC systems in the presence of impulsive noise

Power line communication(PLC)has the potential to become the preferred technique for providing broadband communication to homes and offices with advantage of eliminating the need for new wiring infrastructure and reducing the cost.But it suffers from the impulsive noise because it introduces significant time variance into the power line channel.In this paper,a polar codes based orthogonal frequency division multiplexing(OFDM)PLC system is proposed to deal with the impulsive noise and thereby improve the transmission performance.Firstly,the impulsive noise is modelled with a multi-damped sine function by analyzing the time behavior of impulse events.Then the polar codes are used to combat the impulsive noise of PLC channel,and a low complexity bit-flipping decoding method based on CRC-aided successive cancellation list(CA-SCL)decoding algorithm is proposed.Simulations evaluate the proposed decoding algorithm and the results validate the suggested polar codes based OFDM-PLC scheme which can improve the BER performance of PLC with impulsive interference.

A Study of Radiation-Induced Telomere Instability Using Multiplex Ligation-Dependent Probe Amplification (MLPA)

The integrity of the chromosomes for two WIL2-derived lymphoblastoid cell lines (TK6 and WTK1) in the presence and absence of ionizing radiation was analyzed by Multiplex Ligation-Dependent Probe Amplification (MLPA). The TK6 cell line has the native p53 tumor-suppressor gene, whereas WTK1 cells contain a p53 mutation. Each cell line was isolated pre- and post-irradiation (2 and 3 Gy) and analyzed by MLPA. The impact of irradiation on these two cell lines was investigated using probes that target specific regions on chromosomes associated with subtelomeric regions. Results indicate that WTK1 and TK6 are impacted differently after irradiation, and that each cell line presents its own unique MLPA profile. The most notable differences are the appearance of a number of probes in the post-irradiated MLPA profile that are not present in the controls, and two unique probe signals only seen in WTK1 cells. These results build on our previous studies that indicate how different human cell lines can be affected by radiation in significantly different ways depending on the presence or absence of wild type p53.

Development of a multiplex polymerase chain reaction assay for detection of hepatitis C virus,hepatitis B virus,and human immunodeficiency virus 1

BACKGROUND Hepatitis C virus(HCV),hepatitis B virus(HBV),and human immunodeficiency virus 1(HIV-1)are the most epidemic blood-borne viruses,posing threats to human health and causing economic losses to nations for combating the infection transmission.The diagnostic methodologies that depend on the detection of viral nucleic acids are much more expensive,but they are more accurate than sero-logical testing.AIM To develop a rapid,cost-effective,and accurate diagnostic multiplex polymerase chain reaction(PCR)assay for simultaneous detection of HCV,HBV,and HIV-1.METHODS The design of the proposed PCR assay targets the amplification of a short conserved region featured with a distinguishable melting profile and electro-phoretic molecular weight inside each viral genome.Therefore,this diagnostic method will be appropriate for application in both conventional(combined with electrophoresis)and real-time PCR facilities.Confirmatory in silico investigations were conducted to prove the capability of the approached PCR assay to detect variants of each virus.Then,Egyptian isolates of each virus were subjected to the wet lab examination using the given diagnostic assay.RESULTS The in silico investigations confirmed that the PCR primers can match many viral variants in a multiplex PCR assay.The wet lab experiment proved the efficiency of the assay in distinguishing each viral type through high-resolution melting analysis.Compared to related published assays,the proposed assay in the current study is more sensitive and competitive with many expensive PCR assays.CONCLUSION This study provides a simple,cost-effective,and sensitive diagnostic PCR assay facilitating the detection of the most epidemic blood-borne viruses;this makes the proposed assay promising to be substitutive for the mistakable and cheap serological-based assays.

Real-Time 4-Mode MDM Transmission Using Commercial 400G OTN Transceivers and All-Fiber Mode Multiplexers

Weakly-coupled mode division multiplexing(MDM)technique is considered a promising candidate to enhance the capacity of an optical transmission system,in which mode multiplexers/demultiplexers(MMUX/MDEMUX)with low insertion loss and modal crosstalk are the key components.In this paper,a low-modal-crosstalk 4-mode MMUX/MDEMUX for the weakly-coupled triple-ring-core few-mode fiber(TRC-FMF)is designed and fabricated with side-polishing processing.The measurement results show that a pair of MMUX/MDEMUX and 25 km weakly-coupled TRC-FMF MDM link achieve low modal crosstalk of lower than−17.5 dB and insertion loss of lower than 11.56 dB for all the four modes.Based on the TRC-FMF and all-fiber MMUX/MDEMUX,an experiment for 25 km real-time 4-mode 3-λwavelength division multiplexing(WDM)-MDM transmission is conducted using commercial 400G optical transport network(OTN)transceivers.The experimental results prove weakly-coupled MDM techniques facilitate a smooth upgrade of the optical transmission system.

Study on Distribution of Four Pseudomonas Species in Living Environment Using Multiplex PCR

Purpose: The genus Pseudomonas is a ubiquitous microorganism frequently detected from immunocompromised patients. The inherent resistance to numerous antimicrobial agents contributes to the opportunistic character of this pathogen exhaustive monitoring of this pathogen is considered of critical importance to public health organizations. The reliable identification method able to distinguish genetic close Pseudomonas species is needed, because these organisms are difficult to differentiate by phenotypic or biochemical methods. The purpose of the present study was to design species-specific primers in order to identify and detect four Pseudomonas species which are frequently detected from the human oral cavities, and to investigate the distribution of these organisms in the living environment using a multiplex PCR. Methods: Polymerase chain reaction (PCR) primers were designed based on partial sequences of the rpoD gene of four Pseudomonas species. Swab samples were collected from fifty washstands, and the distribution of Pseudomonas species was investigated using a conventional PCR at genus level and a multiplex PCR at species level. Results: Multiplex PCR method developed in this study was able to distinguish four Pseudomonas species clearly. The genus Pseudomonas was detected from all samples (100%), whereas P. putida, P, aeruginosa, P. stutzeri and P. fluorescens were detected at 44%, 8%, 4% and 2% in fifty swab samples, respectively. Conclusion: Our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and works without requiring DNA extraction. It was indicated that washstands were the uninhabitable environment for P. putida, P, aeruginosa, P. stutzeri and P. fluorescens.

Multiplex Rapid Test with Acceptable Diagnosis Performance as a Solution to Increase Diagnosis of Hepatitis B and C Viruses in Pregnant Women in an Area of High Prevalence of Both Hepatitis Viruses Associated with HIV

Background and Objective: HIV, hepatitis B virus (HBV) and hepatitis C virus (HCV) are very widespread in the world, however, less than 20% of the people affected are diagnosed and treated. This study aimed to determine the prevalence of HIV, HCV and HBV co-infections in pregnant women at Bangui Community University Hospital and the cost of screening. Methods: A cross-sectional study involving consenting pregnant women who came for antenatal care was performed. HIV, HCV antibodies and HBV antigens were detected using Exacto Triplex? HIV/HCV/HBsAg rapid test, cross-validated by ELISA tests. Sociodemographic and professional data, the modes of transmission and prevention of HIV and both hepatitis viruses were collected in a standard sheet and analyzed using the Epi-Info software version 7. Results: Pregnant women aged 15 to 24 were the most affected (45.3%);high school girls (46.0%), and pregnant women living in cohabitation (65.3%) were the most represented. Twenty-five (16.7%) worked in the formal sector, 12.7% were unemployed housewives and the remainder in the informal sector. The prevalence of HIV, HBV, and HCV viruses was 11.8%, 21.9% and 22.2%, respectively. The prevalence of co-infections was 8.6% for HIV-HBV, 10.2% for HIV-HCV, 14.7% for HBV-HCV and 6.5% for HIV-HBV-HCV. All positive results and 10% of negative results by the rapid test were confirmed by ELISA tests. The serology of the three viruses costs 39,000 FCFA (60 Euros) by ELISA compared to 10,000 FCFA (15.00 Euros) with Exacto Triplex? HIV/HCV/AgHBs (BioSynex, Strasbourg, France). Conclusion: The low level of education and awareness of hepatitis are barriers to development and indicate the importance of improving the literacy rate of women in the Central African Republic (CAR). Likewise, the high prevalence of the three viruses shows the need for the urgent establishment of a national program to combat viral hepatitis in the CAR.

Turbulence mitigation scheme based on multiple-user detection in an orbital-angular-momentum multiplexed system

Atmospheric turbulence（AT） induced crosstalk can significantly impair the performance of a free-space optical（FSO）communication link using orbital angular momentum（OAM） multiplexing.In this paper,we propose a multiple-user detection（MUD） turbulence mitigation scheme in an OAM-multiplexed FSO communication link.First,we present a MUD equivalent communication model for an OAM-multiplexed FSO communication link under AT.In the equivalent model,each input bit stream represents one user＇s information.The deformed OAM spatial modes caused by AT,instead of the pure OAM spatial modes,are used as information carriers,and the overlapping between the deformed OAM spatial modes are computed as the correlation coefficients between the users.Then,we present a turbulence mitigation scheme based on MUD idea to enhance AT tolerance of the OAM-multiplexed FSO communication link.In the proposed scheme,the crosstalk caused by AT is used as a useful component to deduce users＇ information.The numerical results show that the performance of the OAM-multiplexed communication link has greatly improved by the proposed scheme.When the turbulence strength C_n^2 is 1 × 10^（-15） m^（-2/3）,the transmission distance is 1000 m and the channel signal-to-noise ratio（SNR）is 26 dB,the bit-error-rate（BER） performance of four spatial multiplexed OAM modes l_m = ＋ 1,＋2,＋3,＋4 are all close to 10-5,and there is a 2-3 fold increase in the BER performance in comparison with those results without the proposed scheme.In addition,the proposed scheme is more effective for an OAM-multiplexed FSO communication link with a larger OAM mode topological charge interval.The proposed scheme is a promising direction for compensating the interference caused by AT in the OAM-multiplexed FSO communication link.

The Difficulties and Mechanism Innovation of Constructing China's Rural Multiplex Financial System

This paper expounds the necessity of constructing China's rural multiplex financial system as follows.First,overcome the bottleneck effect arising from the shortage of capital in the process of development of rural small and medium-sized enterprise,and increase farmers' income;second,provide capital support for rural modern industrial development,and promote rural economic development;third,rationally allocate capital,master the money flow and coordinate urban-rural development.The paper also profoundly analyzes the difficulties of constructing China's rural multiplex financial system as follows.First,the existing rural financial system structure is irrational and the function is unsound;second,non-governmental financial market is disorderly and the organization is unsound;third,the rural financial system is simple and the overhaul of rural credit cooperatives system has not yet been done;fourth,the construction of rural system of financial laws and regulations is laggard,and the construction of rural multiplex financial system lacks good social environment.The specific thoughts,measures and countermeasures of constructing mechanism innovation of China's rural multiplex financial system are put forward as follows.First,increasingly perfect legal system and provide legal support for constructing China's rural multiplex financial system;second,quicken the overhaul of rural credit cooperatives system and perfect its function in China's rural multiplex financial system;third,strengthen the degree of support and development,and change the situation of simple structure of rural financial system;fourth,reinforce administrative supervision and perfect public services,and standardize the rural financial market order.

Establishment of a Multiplex PCR System to Diagnose Tuberculosis and Other Bacterial Infections

In order to rapidly diagnose and differentiate tuberculosis from other bacterial infections, a 16S rRNA gene (16s rDNA)-directed multiplex PCR system was developed. In this system, a pair of universal primers and a tubercle bacillus (Tb)-specific primer were designed based on highly con- served regions and Tb species-specific variable region of bacterial 16s rDNA. A 360bp fragment was detected in all bacteria tested, and a 210bp fragment was found only in Tb. 19 species of known bac- teria including Tb were used for evaluating specificity, universality and sensitivity of the PCR. Candi- da albicans and human diploid cell served as controls. It was found that both 210bp and 360bp frag- ments were amplified only in Tb, and only 360 bp fragment was detected in other 18 species of gener- al bacteria. Candida albicans and human cells were negative for both 360bp and 2l0bp fragments. The lowest detectable level of the PCR was 10 fg of DNA for Escherichia coli and 100 fg of DNA for Tb. The results indicated that this multiplex PCR system for the simultaneous detection of Tb and other common bacteria had higher specificity and sensitivity, as well as good universality and might be useful to rapidly diagnose bacterial infections and effectively distinguish tuberculosis from other bacterial involvement.

Exploiting the Faster-Than-Nyquist Concept in Wavelength-Division Multiplexing Systems Using Duobinary Shaping

This paper begins with Nyquist wavelengthdivision multiplexing （WDM） and then introduces fasterthanNyquist. In fasterthanNyquist a certain amount of intersymbol interference （ISI） is accepted, which violates the fundamental principle of Nyquist WDM. This results in muchrelaxed transceiver bandwidth and simpler spectral design. However, in fasterthanNyquist, implementation complexity is shifted from the transmitter side to the receiver side. Therefore, successful application of fasterthanNyquist depends on innovation in the receiver structure. In this paper, we discuss the guidelines for implementing suboptimum, lowcomplexity receivers based on fasterthanNyquist. We suggest that duobinary shaping is a good technique for trading off achievable spectral efficiency, detection performance, and implementation complexity and might be preferable to Nyquist WDM. Experiments are conducted to verify robustness of the proposed technique.

Time-sequential color code division multiplexing holographic display with metasurface

Color metasurface holograms are powerful and versatile platforms for modulating the amplitude,phase,polarization,and other properties of light at multiple operating wavelengths.However,the current color metasurface holography can only realize static manipulation.In this study,we propose and demonstrate a multiplexing metasurface technique combined with multiwavelength code-division multiplexing(CDM)to realize dynamic manipulation.Multicolor code references are utilized to record information within a single metasurface and increase the information capacity and security for anticracks.A total of 48 monochrome images consisting of pure color characters and multilevel color video frames were reconstructed in dual polarization channels of the birefringent metasurface to exhibit high information density,and a video was displayed via sequential illumination of the corresponding code patterns to verify the ability of dynamic manipulation.Our approach demonstrates significant application potential in optical data storage,optical encryption,multiwavelengthversatile diffractive optical elements,and stimulated emission depletion microscopy.

Flat multifunctional liquid crystal elements through multi-dimensional information multiplexing

Flat optical elements have attracted enormous attentions and act as promising candidates for the next generation of optical components.As one of the most outstanding representatives,liquid crystal(LC)has been widely applied in flat panel display industries and inspires the wavefront modulation with the development of LC alignment techniques.However,most LC elements perform only one type of optical manipulation and are difficult to realize the multifunctionality and light integration.Here,flat multifunctional liquid crystal elements(FMLCEs),merely composed of anisotropic LC molecules with space-variant orientations,are presented for multichannel information manipulation by means of polarization,space and wavelength multiplexing.Specifically,benefiting from the unique light response with the change of the incident polarization,observation plane,and working wavelength,a series of FMLCEs are demonstrated to achieve distinct near-and far-field display functions.The proposed strategy takes full advantage of basic optical parameters as the decrypted keys to improve the information capacity and security,and we expect it to find potential applications in information encryption,optical anti-counterfeiting,virtual/augmented reality,etc.

Planar peristrophic multiplexing metasurfaces

As a promising counterpart of two-dimensional metamaterials,metasurfaces enable to arbitrarily control the wavefront of light at subwavelength scale and hold promise for planar holography and applicable multiplexing devices.Nevertheless,the degrees of freedom(DoF)to orthogonally multiplex data have been almost exhausted.Compared with state-of-theart methods that extensively employ the orthogonal basis such as wavelength,polarization or orbital angular momentum,we propose an unprecedented method of peristrophic multiplexing by combining the spatial frequency orthogonality with the subwavelength detour phase principle.The orthogonal relationship between the spatial frequency of incident light and the locally shifted building blocks of metasurfaces can be regarded as an additional DoF.We experimentally demonstrate the viability of the multiplexed holograms.Moreover,this newly-explored orthogonality is compatible with conventional DoFs.Our findings will contribute to the development of multiplexing metasurfaces and provide a novel solution to nanophotonics,such as large-capacity chip-scale devices and highly integrated communication.

A TDMA Approach for OFDM-Based Multiuser RadCom Systems

This paper is focused on the multiuser implementation of fusion of radar and communication(RadCom)in internet-of-vehicles(IoV)scenarios.Traditional time-division multiple access(TDMA)technology degrades the velocity detection performance of orthogonal frequency-division multiplexing(OFDM)-based RadCom systems.We propose a new TDMA approach for OFDM-based RadCom systems,where multiuser communication and radar detection are completed synchronously.We consider a continuous-wave TDMA OFDM structure in which random user data or Zadoff-Chu(ZC)sequences are transmitted in one symbol duration to ensure detection performance.As an application of interference cancellation method,user data demodulation and environment sensing can be simultaneously accomplished by our proposed approach.We carry out numerical evaluation and show wireless communication and radar detection performance over the continuous-wave TDMA OFDM-based RadCom approach.