Light Scattering Method for Aerosol Sizing Based on Machine Learning.

Optical scattering has been widely used for aerosol sizing due to its noninvasive and real-time measurement. However, it is crucial to retrieve the particle size distribution (PSD) of aerosols without prior knowledge of the refractive index. Now, it has been a great challenge to measure the refractive index in situ. In this study, a novel PSD sensing method utilizing the light scattering angular spectrum (LSAS) and machine learning techniques is proposed to address this challenge. The complex nonlinear relationship between LSAS and PSD can be constructed while accounting for the refractive index of aerosols. A miniaturized prototype sensor is designed and tested on different sizes of aerosol samples. The experiment results showed that the maximum Kullback-Leibler divergence (DKL) of PSD is 0.07, which indicates that the sensing method can provide the ability for highly accurate aerosol PSD measurement without requiring prior knowledge of the refractive index. The compacted prototype sensor shows great potential for aerosol analysis in conventional field measurements outside the laboratory.

Efficient photoelectrocatalytic degradation of pollutants over hydrophobic carbon felt loaded with Fe-doped porous carbon nitride via direct activation of molecular oxygen.

Developing a photoelectric cathode capable of efficiently activating molecular oxygen to degrade pollutants is a coveted yet challenging goal. In pursuit of this, we synthesize a Fe doped porous carbon nitride catalyst (Fe-CN) using an ionothermal strategy and subsequently loaded it on the hydrophobic carbon felt (CF) to fabricate the Fe-CN/CF photoelectric cathode. This cathode benefits from the synergistic effects between the porous CN support and the highly dispersed Fe species, which enhance O2 absorption and activation. Additionally, the hydrophobic CF serves as a gas diffusion layer, accelerating O2 mass transfer. These features enable the Fe-CN/CF cathode to demonstrate notable photoelectrocatalytic (PEC) degradation efficiency. Specifically, under optimal conditions (cathodic bias of -0.3 VAg/AgCl, pH 7, and a catalyst loading of 3 mg/cm2), the system achieves a 76.4% removal rate of tetracycline (TC) within 60 min. The general application potential of this system is further underscored by its ability to remove approximately 98% of 4-chlorophenol (4-CP) and phenol under identical conditions. Subsequent investigations into the active species and degradation pathways reveal that 1O2 and h+ play dominant role during the PEC degradation process, leading to gradually breakdown of TC into less toxicity, smaller molecular intermediates. This work presents a straightforward yet effective strategy for constructing efficient PEC systems that leverage molecular oxygen activation to degrade pollutants.

Real-time synchronized monitoring for the overlap accuracy of the combining beam spot in a wavelength beam combination based on confocal planes.

Beam overlap accuracy in a wavelength beam combination system determines the beam quality and efficiency, so systematic monitoring of overlap accuracy is essential. In this work, a method of performing real-time synchronized monitoring and recording overlap accuracy for a combining beam spot is proposed. Firstly, theoretical calculations for monitoring different wavelength sub-beam positions and angular errors are established. Then, an optical design and grayscale centroid algorithm are developed to analyze and simulate the combination spots. A monitoring device was designed and constructed to meet the requirements of combining system applications, which achieved an accuracy of 8.86 µrad. Finally, the method successfully monitored the system spot fluctuation range within ±22 µrad. This study resolves the issue of distinguishing the different wavelength sub-beams and their response delays in traditional combining beams. It offers precise error data for real-time synchronized calibration of the overlap accuracy in laser beam combining technology.

Towards the Clinical Translation of 3D PLGA/ß-TCP/Mg Composite Scaffold for Cranial Bone Regeneration.

Recent years have witnessed the rapid development of 3D porous scaffolds with excellent biocompatibility, tunable porosity, and pore interconnectivity, sufficient mechanical strength, controlled biodegradability, and favorable osteogenesis for improved results in cranioplasty. However, clinical translation of these scaffolds has lagged far behind, mainly because of the absence of a series of biological evaluations. Herein, we designed and fabricated a composite 3D porous scaffold composed of poly (lactic-co-glycolic) acid (PLGA), ß-tricalcium phosphate (ß-TCP), and Mg using the low-temperature deposition manufacturing (LDM) technique. The LDM-engineered scaffolds possessed highly porous and interconnected microstructures with a porosity of 63%. Meanwhile, the scaffolds exhibited mechanical properties close to that of cancellous bone, as confirmed by the compression tests. It was also found that the original composition of scaffolds could be maintained throughout the fabrication process. Particularly, two important biologic evaluations designed for non-active medical devices, i.e., local effects after implantation and subchronic systemic toxicity tests, were conducted to evaluate the local and systemic toxicity of the scaffolds. Additionally, the scaffolds exhibited significant higher mRNA levels of osteogenic genes compared to control scaffolds, as confirmed by an in vitro osteogenic differentiation test of MC3T3-E1 cells. Finally, we demonstrated the improved cranial bone regeneration performance of the scaffolds in a rabbit model. We envision that our investigation could pave the way for translating the LDM-engineered composite scaffolds into clinical products for cranial bone regeneration.

The opposite effect of tapinarof between IMQ and IL-23 induced psoriasis mouse models.

Tapinarof is an aryl hydrocarbon receptor (AHR) ligand which is used to treat plaque psoriasis in adults. However, the underlying mechanism is not yet fully understood. In this study, we applied two of the most studied psoriasis mouse models: topical application of imiquimod (IMQ) and subcutaneous injection of IL-23. Although both models successfully induced psoriasis-like lesions in mice, tapinarof had a completely opposite effect on the two models. Tapinarof decreased the expression of multiple essential cytokines involved in the pathological IL-23/IL-17/IL-22 axis and ameliorated IMQ-induced psoriatic dermatitis, inhibiting keratinocyte proliferation and abnormal differentiation. However, in the IL-23-injection-model, tapinarof instead aggravated the disease. Here, tapinarof increased epidermal thickness and differentiated epidermal dysplasia in mice. Our data suggest that tapinarof may have different effects on varied types of psoriasis.

Naturally derived highly resilient and adhesive hydrogels with application as surgical adhesive.

The inadequacy of conventional surgical techniques for wound closure and repair in soft and resilient tissues may lead to poor healing outcomes such as local tissue fibrosis and contracture. Therefore, the development of adhesive and resilient hydrogels that can adhere firmly to irregular and dynamic wound interfaces and provide a "tension-free proximity" environment for tissue regeneration has become extremely important. Herein, we describe an integrated modeling-experiment-application strategy for engineering a promising hydrogel-based bioadhesive based on recombinant human collagen (RHC) and catechol-modified hyaluronic acid (HA-Cat). Molecular modeling and simulations were used to verify and explore the hypothesis that RHC and HA-Cat can form an assembly complex through physical interactions. The complex was synergistically crosslinked via a catechol/o-quinone coupling reaction and a carbodiimide coupling reactions, resulting in superior hydrogels with strong adhesion and resilience properties. The application of this bioadhesive to tissue adhesion and wound sealing in vivo was successfully demonstrated, with an optimum collagen index, epidermal thickness, and lowest scar width. Furthermore, subcutaneous implantation demonstrated that the bioadhesive exhibited good biocompatibility and degradability. This newly developed hydrogel may be a highly promising surgical adhesive for medical applications, including wound closure and repair.

S-alkyl Dithiocarbamates Synthesis through Electrochemical Multicomponent Reaction of Thiols, Hydrogen Sulfide, and Isocyanides.

Dithiocarbamates synthesis is extremely important in plenty of biomedical and agrochemical applications, especially fungicide development, but remains a great challenge. In this work, we have successfully developed a multicomponent reaction protocol to convert H2S into S-alkyl dithiocarbamates under constant current conditions. No additional oxidants nor additional catalysts are required, and due to mild conditions, the reactions display a broad substrate scope, including varieties of thiols or disulfides.

Identification of a novel RSRC1-ALK (R6: A20) fusion using next-generation sequencing technique.

Non-small-cell lung cancer (NSCLC) patients with anaplastic lymphoma kinase (ALK) fusion showed promising responses to ALK tyrosine kinase inhibitors (TKIs). In this study, fluorescence in situ hybridization (FISH), immunohistochemistry (IHC), next generation sequencing (NGS) and Sanger sequencing were performed to identify the presence of ALK fusion, to investigate whether the patient may benefit from TKI therapy. Postoperative pathological analysis indicated invasive adenocarcinoma with mainly mucinous type and partial micropapillary type in left lower lung. Minimally invasive adenocarcinoma was seen in left upper lung, with mainly acinar type. NGS detected a novel RSRC1-ALK (R6: A20) fusion in left lower lobe sample, which was presented as the fusion of exon 6 of RSRC1 and exon 20 of ALK gene. Sanger sequencing validated the fusion. Break rearrangement signal of ALK gene was detected in 80% of tumor cells. Immunohistochemistry showed ALK positive expression in lung. For the treatment, the patient received ensartinib hydrochloride with a dose of 225 mg per day. He was in a state of progression-free survival for at least 24 months in follow-up with no complications. NGS can be used for exploring treatment options for NSCLC patients with ALK fusion.

Integrated Human Skin Bacteria Genome Catalog Reveals Extensive Unexplored Habitat-Specific Microbiome Diversity and Function.

The skin is the largest organ in the human body. Various skin environments on its surface constitutes a complex ecosystem. One of the characteristics of the skin micro-ecosystem is low biomass, which greatly limits a comprehensive identification of the microbial species through sequencing. In this study, deep-shotgun sequencing (average 21.5 Gigabyte (Gb)) from 450 facial samples and publicly available skin metagenomic datasets of 2069 samples to assemble a Unified Human Skin Genome (UHSG) catalog is integrated. The UHSG encompasses 813 prokaryotic species derived from 5779 metagenome-assembled genomes, among which 470 are novel species covering 20 phyla with 1385 novel assembled genomes. Based on the UHSG, the core functions of the skin microbiome are described and the differences in amino acid metabolism, carbohydrate metabolism, and drug resistance functions among different phyla are identified. Furthermore, analysis of secondary metabolites of the near-complete genomes further find 1220 putative novel secondary metabolites, several of which are found in previously unknown genomes. Single nucleotide variant (SNV) reveals a possible skin protection mechanism: the negative selection process of the skin environment to conditional pathogens. UHSG offers a convenient reference database that will facilitate a more in-depth understanding of the role of skin microorganisms in the skin.

Differential genome-wide associated variants and enriched pathways of ECG parameters among people with versus without HIV.

OBJECTIVES: People with HIV (PWH) are more likely to develop ECG abnormalities. Substantial evidence exists for genetic contribution to ECG parameters among general population. However, whether and how would host genome associate with ECG parameters among PWH is unclear. Our research aims to analyze and compare genetic variants, mapped genes, and enriched pathways of ECG parameters among PWH and HIV-negative controls. DESIGN: A cross-sectional study. METHOD: We performed a large original genome-wide association study (GWAS) of ECG parameters among PWH ( n  = 1730) and HIV-negative controls ( n =  3746). Genome-wide interaction analyses were also conducted. RESULTS: A total of 18 novel variants were detected among PWH, six for PR interval including rs76345397 at ATL2 , 11 for QRS duration including rs10483994 at KCNK10 and rs2478830 at JCAD , and one for QTc interval (rs9815364). Among HIV-negative controls, we identified variants located at previously reported ECG-related genes ( SCN5A , CNOT1 ). Genetic variants had a significant interaction with HIV infection ( P  < 5 × 10 -8 ), implying that HIV infection and host genome might jointly influence ECG parameters. Mapped genes for PR interval and QRS duration among PWH were enriched in the biological process of viral genome replication and host response to virus, respectively, whereas enriched pathways for PR interval among HIV-negative controls were in the cellular component of voltage-gated sodium channel complex. CONCLUSION: The present GWAS indicated a distinctive impact of host genome on quantitative ECG parameters among PWH. Different from HIV-negative controls, host genome might influence the cardiac electrical activity by interfering with HIV viral infection, production, and latency among PWH.

Clinicopathological characteristics and prognosis of young patients aged ≤45 years old with non-small cell lung cancer.

Lung cancer is rare in young people, but the incidence and mortality are on the rise. We retrospectively analyzed the data of young patients aged ≤45 years diagnosed as lung cancer in our hospital from 2014 to 2021. The purpose was to explore the clinicopathological characteristics of young patients, and the risk factors affecting overall survival (OS) time. The results showed that the young patients were mainly female, had no smoking history, asymptomatic at initial diagnosis, with a high proportion of adenocarcinoma and stage I-II. We divided all patients into two groups according to age and found that the proportion of stage I-II in 18-35 years group was significantly higher than that in 36-45 years group (P = 0.021). The main manifestation of tumor was ground glass opacity (GGO) in 18-35 years group, while most showed non-GGO in 36-45 years group (P = 0.003). The proportion of minimally invasive adenocarcinoma was higher in 18-35 years group, while the invasive adenocarcinoma was higher in 36-45 years group (P = 0.004). Univariate analysis showed that asymptomatic, stage I-II, surgery, women, with few or no metastatic organs had longer OS. Multivariate analysis showed that the independent factors affecting the OS of young patients were tumor stage and more metastatic organs.

Unique Tubular BiOBr/g-C3 N4 Heterojunction with Efficient Separation of Charge Carriers for Photocatalytic Nitrogen Fixation.

The industrial ammonia synthesis process consumes a lot of energy and causes serious environmental pollution. As a sustainable approach for ammonia synthesis, photocatalytic nitrogen reduction employing water as the reducing agent has a lot of potential. A simple surfactant-assisted solvothermal method is used to synthesize g-C3 N4 nanotubes with flower-like spherical BiOBr grown inside and outside (BiOBr/g-C3 N4 , BC). The hollow tubular structure realizes the full use of visible light by the multi-scattering effect of light. Large surface areas and more active sites for N2 adsorption and activation are present in the distinctive spatially dispersed hierarchical structures. Particularly, the quick separation and transfer of electrons and holes are facilitated by the sandwich tubular heterojunctions and tight contact interface of BiOBr and g-C3 N4 . The maximal NH3 generation rate of the BiOBr/g-C3 N4 composite catalysts can reach 255.04 µmol⋅ g-1 ⋅ h-1 , and it is 13.9 and 5.8 times that of pure BiOBr and g-C3 N4 . This work provides a novel method for designing and constructing unique heterojunctions for efficient photocatalytic nitrogen fixation.

Electrical Stimulation Promotes the Vascularization and Functionalization of an Engineered Biomimetic Human Cardiac Tissue.

The formation of multiscale vascular networks is essential for the in vitro construction of large-scale biomimetic cardiac tissues/organs. Although a variety of bioprinting processes have been developed to achieve the construction of mesoscale and large-scale blood vessels, the formation of microvascular networks still mainly depends on the self-assembly behavior of endothelial cells (ECs), which is inefficient and demanding without appropriate stimulus. To address this problem, the elongation and connection of endothelial cells in engineered cardiac tissue (ECT) are sought to promote by electrical stimulation (ES) to achieve vascularization. As proof of the concept, bio-inks are composed of GelMA/fibrin hydrogel, human pluripotent stem cells induced cardiomyocytes (iPSC-CM), and human umbilical vein endothelial cells (HUVEC) are used for the bioprinting of ECTs. It is demonstrated that electrical stimulation significantly promotes the elongation, migration, and interconnection of HUVECs in ECT and increases the expression of related genes. Moreover, ES also enhances the secretion of signal factors interacting between CMs and HUVECs. It seems that the HUVECs further strengthen the contractility of cardiac tissue. Taken together, electrical stimulation promotes vascularization and CMs functionalization in ECT, which has important application potential in the fabrication of vascularized ECT and its clinical transplantation.

Adipose Tissue and Adipose-Tissue-Derived Cell Therapies for the Treatment of the Face and Hands of Patients Suffering from Systemic Sclerosis.

Adipose tissue is recognized as a valuable source of cells with angiogenic, immunomodulatory, reparative and antifibrotic properties and emerged as a therapeutic alternative for the regeneration and repair of damaged tissues. The use of adipose-tissue-based therapy is expanding in autoimmune diseases, particularly in Systemic Sclerosis (SSc), a disease in which hands and face are severely affected, leading to disability and a decrease in quality of life. Combining the advantage of an abundant supply of fat tissue and a high abundance of stem/stromal cells, fat grafting and adipose tissue-derived cell-based therapies are attractive therapeutic options in SSc. This review aims to synthesize the evidence to determine the effects of the use of these biological products for face and hands treatment in the context of SSc. This highlights several points: the need to use relevant effectiveness criteria taking into account the clinical heterogeneity of SSc in order to facilitate assessment and comparison of innovative therapies; second, it reveals some impacts of the disease on fat-grafting success; third, an important heterogeneity was noticed regarding the manufacturing of the adipose-derived products and lastly, it shows a lack of robust evidence from controlled trials comparing adipose-derived products with standard care.

Next-Generation Sequencing of the Complete Huaibei Grey Donkey Mitogenome and Mitogenomic Phylogeny of the Equidae Family.

The Huaibei grey donkey (HGD) is an endangered species and a vital native breed in Anhui Province, China. However, its complete mitogenome, phylogeny, and maternal origin remain unclear. The objectives of this study were to detect the genetic diversity of the HGD and investigate its phylogenetic relationship with other breeds to inform conservation management. The complete mitogenome of the HGD was sequenced through next-generation sequencing, and the most variable region in the mitochondrial DNA displacement-loop (D-loop) was amplified via a polymerase chain reaction (PCR). Next, we used the median-joining network (MJN) to calculate the genetic relationships among populations and the neighbor-jointing method to build a phylogenetic tree and speculate as to its origin. The results showed that the mitogenome contains 22 tRNAs, 2 rRNAs, 13 PCGs, and 1 D-loop region. Analyzing the D-loop region of the HGDs, we identified 23 polymorphic sites and 11 haplotypes. The haplotype and nucleotide diversity were 0.87000 (Hd) and 0.02115 (Pi), respectively. The MJN analysis indicated that the HGD potentially has two maternal lineages, and phylogenetic analysis indicated that the Somali lineage could be the most probable domestication center for this breed. Therefore, our mitogenome analysis highlights the high genetic diversity of the HGD, which may have originated from the Somali wild ass, as opposed to the Asian wild ass. This study will provide a useful resource for HGD conservation and breeding.

Constructed wetlands as hotspots of antibiotic resistance genes and pathogens: Evidence from metagenomic analysis in Chinese rural areas.

In rural China, many constructed wetlands (CWs) have been developed to treat rural wastewater sustainably. However, due to the scarce information on those rural CWs, it is difficult to analyze the biological contaminants within those systems, such as antibiotic resistance genes (ARGs) and pathogens. Based on the data collected from two pilot-scale, one-year-observed CWs, for the first time, this study explored the accumulation of ARGs and pathogens using the metagenomic sequencing approach and SourceTracker analysis under different hydraulic loading rates. The Shannon index of ARGs in the effluent surpassed the level found in the influent. The DESeq2 analysis showed that up to 21.49% of the total pathogen species had increased relative abundance in the effluent compared with the influent. By combining the contribution of substrate and rhizosphere, the CW became a more influencing factor for ARGs and pathogens contamination than the influent. The network analysis revealed a critical but latent fact that the development of antibiotic-resistant pathogens is highly likely to be triggered by the co-occurrence of ARGs and pathogens. Collectively, from the aspect of biological risk, our study showed that CWs alone might not be an ideal solution for improving wastewater treatment in rural China.

Rapid analysis the type of customs paper using Micro-NIR spectrometers and machine learning algorithms.

Quick identification of paper types for customs is extremely crucial. Although there are a variety of researches focus on the discrimination of paper, these techniques either require complex preprocessing or large-scale instruments, which are not suitable for customs environments. In this study, we predicted the type of customs paper by using a Micro-NIR spectrometer, and compared the results with Attenuated Total Reflection-Fourier Transform Infrared Spectroscopy (ATR-FTIR). Four different classification algorithms, including linear and non-linear classifiers: K-nearest neighbor (KNN), soft independent modeling of class analogy (SIMCA), partial least squares discriminant analysis (PLS-DA), and least squares-support vector machine (LS-SVM) were employed to classify the type of paper. 20 groups of datasets were selected by Monte Carlo sampling. For Micro-NIR data, the performances of KNN and LS-SVM were outstanding than SIMCA and PLS-DA, with the average accuracies 96.06% and 98.91%, respectively. The outcome of SIMCA and PLS-DA were similar, with the average accuracies 93.00% and 93.97%. Based on the standard derivation, the best stability of models was LS-SVM (1.06%), followed by PLS-DA (1.12%), KNN (1.22%) and SIMCA (3.07%). Compared with ATR-FTIR, the effects of Micro-NIR were better, which were embodies in the better KNN and SIMCA models, and the comparable LS-SVM model. The result demonstrated that the Micro-NIR combined with machine learning algorithms was an effective method to classify the type of customs paper efficiently and quickly, even better than ATR-FTIR.

Toward an intensive understanding of sewer sediment prokaryotic community assembly and function.

Prokaryotic communities play important roles in sewer sediment ecosystems, but the community composition, functional potential, and assembly mechanisms of sewer sediment prokaryotic communities are still poorly understood. Here, we studied the sediment prokaryotic communities in different urban functional areas (multifunctional, commercial, and residential areas) through 16S rRNA gene amplicon sequencing. Our results suggested that the compositions of prokaryotic communities varied significantly among functional areas. Desulfomicrobium, Desulfovibrio, and Desulfobacter involved in the sulfur cycle and some hydrolytic fermentation bacteria were enriched in multifunctional area, while Methanospirillum and Methanoregulaceae, which were related to methane metabolism were significantly discriminant taxa in the commercial area. Physicochemical properties were closely related to overall community changes (p < 0.001), especially the nutrient levels of sediments (i.e., total nitrogen and total phosphorus) and sediment pH. Network analysis revealed that the prokaryotic community network of the residential area sediment was more complex than the other functional areas, suggesting higher stability of the prokaryotic community in the residential area. Stochastic processes dominated the construction of the prokaryotic community. These results expand our understanding of the characteristics of prokaryotic communities in sewer sediment, providing a new perspective for studying sewer sediment prokaryotic community structure.

Genome-wide associated variants of subclinical atherosclerosis among young people with HIV and gene-environment interactions.

BACKGROUND: Genome-wide association studies (GWAS) have identified some variants associated with subclinical atherosclerosis (SCA) in general population but lacking sufficient validation. Besides traditional risk factors, whether and how would genetic variants associate with SCA among people with HIV (PWH) remains to be elucidated. METHOD: A large original GWAS and gene-environment interaction analysis of SCA were conducted among Chinese PWH (n = 2850) and age/sex-matched HIV-negative controls (n = 5410). Subgroup analyses by age and functional annotations of variants were also performed. RESULTS: Different from HIV-negative counterparts, host genome had a greater impact on young PWH rather than the elders: one genome-wide significant variant (rs77741796, P = 2.20 × 10-9) and eight suggestively significant variants (P < 1 × 10-6) were identified to be specifically associated with SCA among PWH younger than 45 years. Seven genomic loci and 15 genes were mapped to play a potential role on SCA among young PWH, which were enriched in the biological processes of atrial cardiac muscle cell membrane repolarization and molecular function of protein kinase A subunit binding. Furthermore, genome-wide interaction analyses revealed significant HIV-gene interactions overall as well as gene-environment interactions with alcohol consumption, tobacco use and obesity among PWH. The identified gene-environment interaction on SCA among PWH might be useful for discovering high-risk individuals for the prevention of SCA, particularly among those with tobacco use and alcohol consumption. CONCLUSION: The present study provides new clues for the genetic contribution of SCA among young PWH and is the starting point of precision intervention targeting HIV-related atherosclerosis.

Complete Mitochondrial Genome, Genetic Diversity and Phylogenetic Analysis of Pingpu Yellow Chicken (Gallus gallus).

In this study, the complete mitochondrial genome sequence of one female Pingpu Yellow chicken (PYC) and the D-loop sequences obtained from 60 chickens were analyzed to investigate their genetic diversity and phylogeny. The total length of the PYC mitogenome is 16,785 bp and that of the complete D-loop is 1231 to 1232 bp. The mitogenome comprises 22 transfer ribonucleic acids (tRNAs), 2 ribosomal ribonucleic acids (rRNAs), 13 protein-coding genes (PCGs), and 1 non-coding control region (D-loop). Additionally, the total length of the 13 PCGs is 11,394 bp, accounting for 67.88% of the complete mitogenome sequence, and the PCGs region has 3798 codons. A majority of the PCGs have ATG as the start codon. The haplotype and nucleotide diversity of PYC were 1.00000 ± 0.00029 and 0.32678 ± 0.29756, respectively. In the D-Loop data set, we found 25 polymorphic sites, which determined 18 haplotypes and 3 major haplogroups (A-C). Therefore, PYC has a classical vertebrate mitogenome, with comparatively high nucleotide diversity and potentially three maternal lineages. The neighbor-joining (NJ) tree analysis results showed PYC grouped with the Luhua (MT555049.1) and Nandan chickens (KP269069.1), which indicates that PYC is closely related to these two breeds.