Cloning of Macaque Monkeys by Somatic Cell Nuclear Transfer.

Generation of genetically uniform non-human primates may help to establish animal models for primate biology and biomedical research. In this study, we have successfully cloned cynomolgus monkeys (Macaca fascicularis) by somatic cell nuclear transfer (SCNT). We found that injection of H3K9me3 demethylase Kdm4d mRNA and treatment with histone deacetylase inhibitor trichostatin A at one-cell stage following SCNT greatly improved blastocyst development and pregnancy rate of transplanted SCNT embryos in surrogate monkeys. For SCNT using fetal monkey fibroblasts, 6 pregnancies were confirmed in 21 surrogates and yielded 2 healthy babies. For SCNT using adult monkey cumulus cells, 22 pregnancies were confirmed in 42 surrogates and yielded 2 babies that were short-lived. In both cases, genetic analyses confirmed that the nuclear DNA and mitochondria DNA of the monkey offspring originated from the nucleus donor cell and the oocyte donor monkey, respectively. Thus, cloning macaque monkeys by SCNT is feasible using fetal fibroblasts.

Heterogeneous nuclear ribonucleoprotein A3 controls mitotic progression of neural progenitors via interaction with cohesin.

Cortex development is controlled by temporal patterning of neural progenitor (NP) competence with sequential generation of deep and superficial layer neurons, but underlying mechanisms remain elusive. Here, we report a role for heterogeneous nuclear ribonucleoprotein A3 (HNRNPA3) in regulating the division of early cortical NPs that mainly give rise to deep-layer neurons via direct neurogenesis. HNRNPA3 is expressed at high levels in NPs of mouse and human cortex at early stages, with a unique peri-chromosome pattern. Intriguingly, downregulation of HNRNPA3 caused chromosome disarrangement, which hindered normal separation of chromosomes during NP division, leading to mitotic delay. Furthermore, HNRNPA3 is associated with the cohesin-core subunit SMC1A and controls its association with chromosomes, implicating a mechanism for the role of HNRNPA3 in regulating chromosome segregation in dividing NPs. Hnrnpa3-deficient mice exhibited reduced cortical thickness, especially of deep layers. Moreover, downregulation of HNRNPA3 in cultured human cerebral organoids led to marked reduction in NPs and deep-layer neurons. Thus, this study has identified a crucial role for HNRNPA3 in NP division and highlighted the relationship between mitosis progression and early neurogenesis.

Autism-like behaviours and germline transmission in transgenic monkeys overexpressing MeCP2.

Methyl-CpG binding protein 2 (MeCP2) has crucial roles in transcriptional regulation and microRNA processing. Mutations in the MECP2 gene are found in 90% of patients with Rett syndrome, a severe developmental disorder with autistic phenotypes. Duplications of MECP2-containing genomic segments cause the MECP2 duplication syndrome, which shares core symptoms with autism spectrum disorders. Although Mecp2-null mice recapitulate most developmental and behavioural defects seen in patients with Rett syndrome, it has been difficult to identify autism-like behaviours in the mouse model of MeCP2 overexpression. Here we report that lentivirus-based transgenic cynomolgus monkeys (Macaca fascicularis) expressing human MeCP2 in the brain exhibit autism-like behaviours and show germline transmission of the transgene. Expression of the MECP2 transgene was confirmed by western blotting and immunostaining of brain tissues of transgenic monkeys. Genomic integration sites of the transgenes were characterized by a deep-sequencing-based method. As compared to wild-type monkeys, MECP2 transgenic monkeys exhibited a higher frequency of repetitive circular locomotion and increased stress responses, as measured by the threat-related anxiety and defensive test. The transgenic monkeys showed less interaction with wild-type monkeys within the same group, and also a reduced interaction time when paired with other transgenic monkeys in social interaction tests. The cognitive functions of the transgenic monkeys were largely normal in the Wisconsin general test apparatus, although some showed signs of stereotypic cognitive behaviours. Notably, we succeeded in generating five F1 offspring of MECP2 transgenic monkeys by intracytoplasmic sperm injection with sperm from one F0 transgenic monkey, showing germline transmission and Mendelian segregation of several MECP2 transgenes in the F1 progeny. Moreover, F1 transgenic monkeys also showed reduced social interactions when tested in pairs, as compared to wild-type monkeys of similar age. Together, these results indicate the feasibility and reliability of using genetically engineered non-human primates to study brain disorders.

Efficient and risk-reduced genome editing using double nicks enhanced by bacterial recombination factors in multiple species.

Site-specific DNA double-strand breaks have been used to generate knock-in through the homology-dependent or -independent pathway. However, low efficiency and accompanying negative impacts such as undesirable indels or tumorigenic potential remain problematic. In this study, we present an enhanced reduced-risk genome editing strategy we named as NEO, which used either site-specific trans or cis double-nicking facilitated by four bacterial recombination factors (RecOFAR). In comparison to currently available approaches, NEO achieved higher knock-in (KI) germline transmission frequency (improving from zero to up to 10% efficiency with an average of 5-fold improvement for 8 loci) and 'cleaner' knock-in of long DNA fragments (up to 5.5 kb) into a variety of genome regions in zebrafish, mice and rats. Furthermore, NEO yielded up to 50% knock-in in monkey embryos and 20% relative integration efficiency in non-dividing primary human peripheral blood lymphocytes (hPBLCs). Remarkably, both on-target and off-target indels were effectively suppressed by NEO. NEO may also be used to introduce low-risk unrestricted point mutations effectively and precisely. Therefore, by balancing efficiency with safety and quality, the NEO method reported here shows substantial potential and improves the in vivo gene-editing strategies that have recently been developed.

Efficient HIE-FDTD method for designing a dual-band anisotropic terahertz absorption structure.

The finite-difference time-domain (FDTD) method is considered to be one of the most accurate and common methods for the simulation of optical devices. However, the conventional FDTD method is subject to the Courant-Friedrich-Levy condition, resulting in extremely low efficiency for calculating two-dimensional materials (2DMs). Recent researches on the hybrid implicit-explicit FDTD (HIE-FDTD) method show that the method can efficiently simulate homogeneous and isotropic 2DMs such as graphene sheet; however, it is inapplicable to the anisotropic medium. In this paper, we propose an in-plane anisotropic HIE-FDTD method to simulate optical devices containing graphene and black phosphorus (BP) sheets. Numerical analysis shows that the proposed method is accurate and efficient. With this method, we present a novel multi-layer graphene-BP-based dual-band anisotropic terahertz absorption structure (GBP-DATAS) and analyze its optical characteristics. Combining the advantages of graphene and BP localized surface plasmons, the GBP-DATAS demonstrates strong anisotropic plasmonic resonance and high absorption rate in the terahertz band.

Refractive index sensor with alternative high performance using black phosphorus in the all-dielectric configuration.

We theoretically propose a nonplasmonic optical refractive index sensor based on black phosphorus (BP) and other dielectric materials in the infrared band. Due to the anisotropic property of BP, the proposed sensor can achieve alternative sensitivity and figure of merit (FOM) in its different crystal directions. The high sensitivity and FOM are attributed to the strong magnetic resonance in the all-dielectric configuration. The coupled-mode theory (CMT) is used to verify the simulation results and reveal the physical mechanism. Furthermore, influences of the sample and the incident angle on the performance of the sensor are also discussed. Our design utilizes a simple dielectric structure with a BP monolayer, which exhibits great potential for the future high-performance sensor with low cost.

Tunable dual-band terahertz absorber with all-dielectric configuration based on graphene.

In this paper, we theoretically design a dual-band graphene-based terahertz (THz) absorber combining the magnetic resonance with a THz cold mirror without any metallic loss. The absorption spectrum of the all-dielectric THz absorber can be actively manipulated after fabrication due to the tunable conductivity of graphene. After delicate optimization, two ultra-narrow absorption peaks are achieved with respective full width at half maximum (FWHM) of 0.0272 THz and 0.0424 THz. Also, we investigate the effect of geometric parameters on the absorption performance. Coupled mode theory (CMT) is conducted on the dual-band spectrum as an analytic method to confirm the validity of numerical results. Furthermore, physical mechanism is deeply revealed with magnetic and electric field distributions, which demonstrate a totally different principle with traditional plasmonic absorber. Our research provides a significant design guide for developing tunable multi-resonant THz devices based on all-dielectric configuration.

Enhanced terahertz modulation using a plasmonic perfect absorber based on black phosphorus.

In this paper, we design a plasmonic perfect absorber based on black phosphorus (BP) with enhanced terahertz modulation. By tuning the chemical potential (µc) of BP, the modulation depth can reach up to 95%. The influence of geometric size and bandgap of BP on reflection spectra is also investigated. Moreover, the effect of the incident angle on the reflectance is discussed with different values of µc. Our results show that the plasmonic nanoslit mode contributes to the enhancement of the modulation effect. This simple periodical structure provides a potential route to design a tunable plasmonic BP-based modulator in the THz range.

Anisotropic infrared plasmonic broadband absorber based on graphene-black phosphorus multilayers.

Two-dimensional materials (2DMs) such as graphene and black phosphorus (BP) have aroused considerable attentions in the past few years. Engineering and enhancing their light-matter interaction is possible due to their support for localized surface plasmon resonances in the infrared regime. In this paper, we have proposed an infrared broadband absorber consisting of multilayer graphene-BP nanoparticles sandwiched between dielectric layers. Benefiting from the properties of graphene and BP, the absorber exhibits both perfect broadband responses and strong anisotropy beyond individual graphene and BP layers. The absorber is tunable with the variation of geometric parameters as well as the doping levels of graphene and BP. The physical insight is revealed by electric field distributions. Furthermore, the angular robustness for incident wave is investigated. The proposed anisotropic omnidirectional broadband absorber may have promising potential applications in various biosensing, communication and imaging systems.

Tunable broadband terahertz absorber based on multilayer graphene-sandwiched plasmonic structure.

We numerically demonstrate a tunable broadband terahertz absorber with near-unity absorption by using multilayer graphene ribbons sandwiched in a plasmonic integrated structure. By stacking slightly different widths of graphene ribbons in a sandwiched configuration, the absorption bandwidth can be increased because of the different resonant modes closely positioned together. The absorption spectrum's center frequency can be manipulated by varying the graphene's chemical potential, which provides a flexible way to design and optimize absorption property after fabrication. Furthermore, the structure can tolerate a wide range of incident angles, while the improved structure with graphene nanoparticles also shows polarization-independent feature. In this routine, stacking more graphene ribbons or particles with well-designed dimensions can further increase the bandwidth, as long as the metamaterial dimension satisfies the sub-wavelength condition. Therefore, our research provides an important theoretical guide for designing various graphene-based tunable broadband absorbers at terahertz, infrared, and microwave frequencies. This may have promising applications in imaging, sensing, and novel optoelectronic devices.

Age-related nomograms of serum anti-Mullerian hormone levels in female monkeys: Comparison of rhesus (Macaca mulatta) and cynomolgus (Macaca fascicularis) monkeys.

AMH is regarded as a promising predictor for ovarian reserve in humans and non-human primate, and widely used in human medicine to predict ovarian response to gonadotropin, menopause and premature ovarian failure. However, large data set on the range of AMH levels in nonhuman primates is still scarce, which limited its applications largely. In this study, age-related AMH nomograms of rhesus (Macaca mulatta) and cynomolgus (Macaca fascicularis) were produced and compared. 219 rhesus and 529 cynomolgus monkeys ranging from infancy to adult were included. In total, the mean serum AMH levels in cynomolgus monkeys were higher than that of rhesus monkeys (14.6 ±â€¯5.3 ng/ml vs 9.5 ±â€¯6.0 ng/ml, P < 0.001). AMH was inversely correlated with age (r = -0.371, P < 0.001) in rhesus, while the negative correlation did not reach statistical significance in cynomolgus monkeys (r = -0.044, P = 0.156). The maximum mean AMH levels were attained at the subgroup of 4-11 yr and the lowest AMH levels were obtained at the subgroup of ≧12 yr in both primates, corresponding to their fertility potential. In rhesus monkeys, from 1 to 11 years old, AMH level remained stable (1-3 yr: ß = 2.784, P = 0.340; 4-11 yr: r = 0.100, P = 0.110) whereas from 12 yr onward, an inverse correlation between AMH and age (r = -0.450, P = 0.02) was observed. Similarly, AMH appeared stable from 1 to 3 yr (ß = -2.289, P = 0.429) and showed an inverse correlation with age (r = -0.521, P < 0.001) from 12 yr onward in cynomolgus monkeys, while a positive correlation was observed (r = 0.156, P = 0.001) from 4 to 11 yr. AMH levels were relatively stable across the menstrual cycle in both primates and no seasonal difference for AMH levels was observed in rhesus monkeys. Body mass index did not affect serum AMH levels in both primates. Our nomograms of serum AMH provide a reference guide on AMH longitudinal distribution by age for Macaca monkeys and might facilitate its applications.

Enhanced expression of Vastatin inhibits angiogenesis and prolongs survival in murine orthotopic glioblastoma model.

BACKGROUND: Antiangiogenic therapies are considered promising for the treatment of glioblastoma (GB). The non-collagenous C-terminal globular NC1 domain of type VIII collagen a1 chain, Vastatin, is an endogenous antiangiogenic polypeptide. Sustained enhanced expression of Vastatin was shown to inhibit tumour growth and metastasis in murine hepatocellular carcinoma models. In this study, we further explored the efficacy of Vastatin in the treatment of GB xenografts. METHOD: Treatment of Vastatin was carried out using a nanopolymer gene vector PEI600-CyD-Folate (H1). Antiangiogenic effect of Vastatin was tested in vitro by using co-culture system and conditioned medium. An orthotopic GB murine model was established to examine the in vivo therapeutic effect of Vastatin alone treatment and its combination with temozolomide. RESULTS: Vastatin gene transfection mediated by H1 could target tumour cells specifically and suppress the proliferation of microvessel endothelial cells (MECs) through a paracrine inhibition manner. Enhancing Vastatin expression by intracerebral injection of H1-Vastatin significantly prolonged animal survival from 48 to 75 days in GB murine model, which was comparable to the effect of Endostatin, the most studied endogenous antiangiogenic polypeptide. The diminished presence of CD34 positive cells in the GB xenografts suggested that Vastatin induced significant antiangiogenesis. Moreover, a synergistic effect in extending survival was detected when H1-Vastatin was administered with temozolomide (TMZ) in GB chemoresistant murine models. CONCLUSION: Our results suggest, for the first time, that Vastatin is an antiangiogenic polypeptide with significant potential therapeutic benefit for GB. H1-Vastatin gene therapy may have important implications in re-sensitizing recurrent GB to standard chemotherapeutic agents.

Serial optical coherence tomography angiography for corneal vascularization.

PURPOSE: Recently, optical coherence tomography angiography (OCTA) has been used to successfully delineate vessels within the retina. This current study aims to assess corneal vascularization secondary to herpetic keratitis pre- and post-treatment using serial OCTA imaging adapted for the anterior segment. METHODS: All eyes were scanned using the split-spectrum amplitude decorrelation angiography (SSADA) algorithm on the AngioVue OCTA system (Optovue Inc. Fremont, CA, USA) with an anterior segment lens adapter. Multiple scans in the regions of interest (ROI) before and after treatment were analysed to assess change in corneal vascularization in response to each treatment modality. RESULTS: We analyzed a total of 12 OCTA scans in three eyes with corneal vascularization, comparing images pre- and 3 months post-treatment. We found that the OCTA was able to detect a significant decrease in area of vascularization in all eyes: including fine-needle diathermy (48 ± 7 to 41 ± 5 %, P = 0.048), subconjunctival bevacizumab (45 ± 7 to 38 ± 5 %, P = 0.015) and systemic steroid treatment following graft rejection (38 ± 1 to 32 ± 2 %, P = 0.003). CONCLUSIONS: Our preliminary study of serial OCTA scans suggests that this may be a useful tool for objective quantification of corneal vascularization. Future development of image processing software will be needed for clinical use or trials to evaluate anti-vascular therapies.

Enhanced spatial near-infrared modulation of graphene-loaded perfect absorbers using plasmonic nanoslits.

Modulating spatial near-infrared light for ultra-compact electro-optic devices is a critical issue in optical communication and imaging applications. To date, spatial near-infrared modulators based on graphene have been reported, but they showed limited modulation effects due to the relatively weak light-graphene interaction. In combination with graphene and metallic nanoslits, we design a kind of ultrathin near-infrared perfect absorber with enhanced spatial modulation effects and independence on a wide range of incident angles. The modulated spectral shift of central wavelength is up to 258.2 nm in the near-infrared range, which is more promising in applications than state-of-the-art devices. The modulation enhancement is attributed to the plasmonic nanoslit mode, in which the optical electric field is highly concentrated in the deep subwavelength scale and the light-graphene interaction is significantly strengthened. The physical insight is deeply revealed by a combination of equivalent circuit and electromagnetic field analysis. The design principles are not only crucial for spatial near-infrared modulators, but also provide a key guide for developing active near-infrared patch nanoantennas based on graphene.

A study of the immune properties of human umbilical cord lining epithelial cells.

BACKGROUND AIMS: Our previous study has demonstrated the stem cell-like properties of human umbilical cord lining epithelial cells (CLECs) and their capability for epidermal reconstitution in organotypic skin culture; however, the immunogenicity of these cells has not been clearly defined. We assessed several aspects of the immune properties of CLECs in vitro. METHODS: We examined CLECs for their immunoregulatory function in a mixed lymphocyte culture experiment. We characterized the expression patterns of the major histocompatibility complex (MHC), co-stimulatory molecules and the pro-/anti-inflammatory cytokines and growth factors in CLECs by means of reverse transcription-polymerase chain reaction, Western blotting, flow cytometry and FlowCytomix multiple analyte detection assays. RESULTS: CLECs were found not to induce but to suppress the proliferation response of the peripheral blood mononuclear cells in a mixed lymphocyte culture assay. They did not express the MHC class II antigen HLA-DR but the non-classic MHC class I antigens HLA-G and HLA-E and lacked the expression of the co-stimulatory molecules CD40, CD80 and CD86. In addition, they produced less interleukin-1ß and transforming growth factor-ß1 but more interleukin-4 and hepatocyte growth factor than did adult keratinocytes, a pattern in favor of wound healing with less inflammation response. CONCLUSIONS: Our data suggest that CLECs have an immunosuppressive function in addition to their low immunogenicity. This could be at least partially explained by their expression of HLA-G and HLA-E molecules associated with immune tolerance and absence of HLA-DR and co-stimulatory molecules. The demonstration that CLECs produce a favorable pattern of cytokines and growth factors for wound healing provides further support for their potential clinical application in allogeneic cell therapy.

How Fe-bearing materials affect soil arsenic bioavailability to rice: A meta-analysis.

Arsenic (As) contamination is widespread in soil and poses a threat to agricultural products and human health due to its high susceptibility to absorption by rice. Fe-bearing materials (Fe-Mat) display significant potential for reducing As bioavailability in soil and bioaccumulation in rice. However, the remediation effect of various Fe-Mat is often inconsistent, and the response to diverse environmental factors is ambiguous. Here, we conducted a meta-analysis to quantitatively assess the effects of As in soils, rice roots, and grains based on 673, 321, and 305 individual observations from 67 peer-reviewed articles, respectively. On average, Fe-Mat reduced As bioavailability in soils, rice roots, and grains by 28.74 %, 33.48 %, and 44.61 %, respectively. According to the analysis of influencing factors, the remediation efficiency of Fe-Mat on As-contaminated soil was significantly enhanced with increasing Fe content in the material, in which the industry byproduct was the most effective in soils (-42.31 %) and rice roots (-44.57 %), while Fe-biochar was superior in rice grains (-54.62 %). The efficiency of Fe-Mat in minimizing soil As mobility was negatively correlated with soil Fe content, CEC, and pH. In addition, applying Fe-Mat in alkaline soils with higher silt, lower clay and available P was more effective in reducing As in rice grains. A higher efficiency of applying Fe-Mat under continuous flooding conditions (27.39 %) compared with alternate wetting and drying conditions (23.66 %) was also identified. Our results offer an important reference for the development of remediation strategies and methods for various As-contaminated paddy soils.

Case report: Pulse cyclophosphamide for treatment of multi-agent-refractory hepatic graft-versus-host disease.

Graft-versus-host disease (GVHD) is a common complication in patients receiving allogeneic hematopoietic stem cell transplantation (HSCT). GVHD is characterized as either acute or chronic based on symptomatology and histopathological findings. Despite advancements in disease-targeting therapeutics, steroid-refractory GVHD remains a significant contributor to mortality in HSCT recipients, highlighting the gaps in our understanding of its pathophysiology and treatment strategies. We present the case of a 46-year-old woman diagnosed with acute undifferentiated leukemia, who exhibited persistently elevated levels of serum total bilirubin (T.Bili), alkaline phosphatase (ALP), and liver function tests (LFTs) beginning on [day +201] post-haploidentical peripheral blood stem cell (PBSC) transplantation. The patient received fludarabine/total body irradiation (Flu/TBI) as a myeloablative conditioning regimen and post-transplant cyclophosphamide/tacrolimus/mycophenolate mofetil (PTCy/Tac/MMF) as GVHD prophylaxis. A liver biopsy confirmed the diagnosis of GVHD, while other possible etiologies were excluded by corresponding tests. Initial treatment with prednisone and tacrolimus, and the later addition of ruxolitinib, all showed poor response indicated by worsening T.Bili, ALP, and LFTs at the same time. Based on a multidisciplinary comprehensive assessment, we decided to administer 1,000 mg/m2 (1,600 mg) of cyclophosphamide ("pulse Cy"), which resulted in a dramatic improvement in T.Bili and transaminases starting from the very next day. A durable response to pulse cyclophosphamide was observed, as all indicators normalized ("complete response") within 55 days without relapses. The patient remains in good health with no recurrence of hepatic GVHD. To our knowledge, this is the first case in which Grade IV hepatic GVHD, refractory to multiple agents including steroids, tacrolimus, and ruxolitinib, demonstrated a complete response to pulse cyclophosphamide. The success highlights the potential therapeutic role of cyclophosphamide, a potent and cost-effective chemotherapy agent, in treating multi-agent-refractory GVHD. Large-scale clinical trials are warranted to validate its efficacy in this setting.

The cost-effectiveness of zolbetuximab in CLDN18.2-positive gastric or gastroesophageal junction adenocarcinoma.

Aim: To estimate the cost-effectiveness of zolbetuximab plus capecitabine/oxaliplatin (CAPOX) in CLDN18.2-positive, HER2-negative, mG/GEJ adenocarcinoma from the perspective of Chinese payers. Materials & methods: A partitioned survival model was developed to assess the costs, quality-adjusted life years (QALYs) and incremental cost-effectiveness ratios (ICER) of zolbetuximab plus CAPOX versus placebo plus CAPOX. Sensitivity analyses were performed to test the robustness of model. Results: Zolbetuximab plus CAPOX gained an additional cost of $91,551 and an extra health benefit of 0.24 QALY over placebo plus CAPOX, producing an ICER of $388,186/QALY, which exceeded the willingness-to-pay threshold of $38,223/QALY. Sensitivity analysis shows that the model was generally robust. Conclusion: Zolbetuximab plus CAPOX would not be a cost-effective first-line treatment regimen in CLDN18.2-positive, HER2-negative, mG/GEJ adenocarcinoma in China.

[Box: see text].

Nano- and microplastics drive the dynamic equilibrium of amoeba-associated bacteria and antibiotic resistance genes.

As emerging pollutants, microplastics have become pervasive on a global scale, inflicting significant harm upon ecosystems. However, the impact of these microplastics on the symbiotic relationship between protists and bacteria remains poorly understood. In this study, we investigated the mechanisms through which nano- and microplastics of varying sizes and concentrations influence the amoeba-bacterial symbiotic system. The findings reveal that nano- and microplastics exert deleterious effects on the adaptability of the amoeba host, with the magnitude of these effects contingent upon particle size and concentration. Furthermore, nano- and microplastics disrupt the initial equilibrium in the symbiotic relationship between amoeba and bacteria, with nano-plastics demonstrating a reduced ability to colonize symbiotic bacteria within the amoeba host when compared to their microplastic counterparts. Moreover, nano- and microplastics enhance the relative abundance of antibiotic resistance genes and heavy metal resistance genes in the bacteria residing within the amoeba host, which undoubtedly increases the potential transmission risk of both human pathogens and resistance genes within the environment. In sum, the results presented herein provide a novel perspective and theoretical foundation for the study of interactions between microplastics and microbial symbiotic systems, along with the establishment of risk assessment systems for ecological environments and human health.