The lymphocyte-specific protein tyrosine kinase (LCK) plays a crucial role in both T-cell development and activation. Dysregulation of LCK signaling has been demonstrated to drive the oncogenesis of T-cell acute lymphoblastic leukemia (T-ALL), thus providing a therapeutic target for leukemia treatment. In this study, we introduced a sophisticated virtual screening strategy combined with biological evaluations to discover potent LCK inhibitors. Our initial approach involved utilizing the PLANET algorithm to assess and contrast various scoring methodologies suitable for LCK inhibitor screening. After effectively evaluating PLANET, we progressed to devise a virtual screening workflow that synergistically combines the strengths of PLANET with the capabilities of Schrödinger's suite. This integrative strategy led to the efficient identification of four potential LCK inhibitors. Among them, compound 1232030-35-1 stood out as the most promising candidate with an IC50 of 0.43 nM. Further in vitro bioassays revealed that 1232030-35-1 exhibited robust antiproliferative effects on T-ALL cells, which was attributed to its ability to suppress the phosphorylations of key molecules in the LCK signaling pathway. More importantly, 1232030-35-1 treatment demonstrated profound in vivo antileukemia efficacy in a human T-ALL xenograft model. In addition, complementary molecular dynamics simulations provided deeper insight into the binding kinetics between 1232030-35-1 and LCK, highlighting the formation of a hydrogen bond with Met319. Collectively, our study established a robust and effective screening strategy that integrates AI-driven and conventional methodologies for the identification of LCK inhibitors, positioning 1232030-35-1 as a highly promising and novel drug-like candidate for potential applications in treating T-ALL.
Persimmon (Diospyros kaki) leaf is widely used as a tea substitute in East Asia, offering potential health benefits. Although studies have highlighted their effects on hyperpigmentation disorders, the active components remain unidentified. This study introduces a novel approach combining LC-MS/MS-based molecular networking with AlphaFold2-enabled virtual screening to expedite the identification of bioactive components in persimmon leaf. A total of 105 compounds were identified by MS/MS analysis. Further, virtual screening identified five flavonoids with potential anti-melanogenic properties. Bioassays confirmed myricetin, quercetin, and kaempferol inhibited melanogenesis in human melanocytes in a dose-dependent manner. Biolayer interferometry assays revealed strong binding affinity between these flavonols and hsTYR, with KD values of 23.26 ± 11.77 for myricetin, 12.43 ± 0.37 for quercetin, and 14.99 ± 3.80 µM for kaempferol. Molecular dynamics simulations provided insights into the binding interactions of these flavonols with hsTYR, particularly highlighting the essential role of the 3-OH group on the C-ring. This study elucidates the bioactive components responsible for the anti-melanogenic effects of persimmon leaf, supporting their use in product development.
Oncogenic overexpression or activation of C-terminal Src kinase (CSK) has been shown to play an important role in triple-negative breast cancer (TNBC) progression, including tumor initiation, growth, metastasis, drug resistance. This revelation has pivoted the focus toward CSK as a potential target for novel treatments. However, until now, there are few inhibitors designed to target the CSK protein. Responding to this, our research has implemented a comprehensive virtual screening protocol. By integrating energy-based screening methods with AI-driven scoring functions, such as Attentive FP, and employing rigorous rescoring methods like Glide docking and molecular mechanics generalized Born surface area (MM/GBSA), we have systematically sought out inhibitors of CSK. This approach led to the discovery of a compound with a potent CSK inhibitory activity, reflected by an IC50 value of 1.6 nM under a homogeneous time-resolved fluorescence (HTRF) bioassay. Subsequently, molecule 2 exhibits strong growth inhibition of MD anderson - metastatic breast (MDA-MB) -231, Hs578T, and SUM159 cells, showing a level of growth inhibition comparable to that observed with dasatinib. Treatment with molecule 2 also induced significant G1 phase accumulation and cell apoptosis. Furthermore, we have explored the explicit binding interactions of the compound with CSK using molecular dynamics simulations, providing valuable insights into its mechanism of action.
The accumulation of ice can pose numerous inconveniences and potential hazards, profoundly affecting both human productivity and daily life. To combat the challenges posed by icing, extensive research efforts have been dedicated to the development of low-ice adhesion surfaces. In this study, we harness the power of molecular dynamics simulations to delve into the intricate dynamics of polymer chains and their role in determining the modulus of the material. We present a novel strategy to prepare ultralow-modulus poly(dimethylsiloxane) (PDMS) elastomers with a molecular brush configuration as icephobic materials. The process involves grafting monohydride-terminated PDMS (H-PDMS) as side chains onto backbone chain PDMS with pendant vinyl functional groups to yield a molecular brush structure. The segments of this polymer structure effectively restrict interchain entanglement, thereby rendering a lower modulus compared to traditional linear structures at an equivalent cross-linking density. The developed soft coating exhibits a remarkably ultralow ice adhesion strength of 13.1 ± 1.1 kPa. Even after enduring 50 cycles of icing and deicing, the ice adhesion strength of this coating steadfastly stayed below 16 kPa, showing no notable increase. Importantly, the molecular brush coating applied to glass demonstrated an impressive light transmittance of 92.1% within the visible light spectrum, surpassing the transmittance of bare glass, which was measured at 91.3%. This icephobic coating with exceptional light transmittance offers a wide range of applications and holds significant potential as a practical icephobic material.
The gram-negative facultative intracellular pathogen Salmonella enterica serotype Choleraesuis, also known as S. Choleraesuis, is a major financial loss for the pig business. C500 is a vaccine strain that has been used for preventing S. Choleraesuis infection in pigs for many years in China. Although it possessed good immunogenicity and protection efficacy, it still showed severe side effects. The truncation of the key gene rpoS in C500 was believed to take the major responsibility for its attenuation. To achieve a good balance between attenuation and immunogenicity, rpoS was restored to an active state, and other essential virulent genes of crp, fur, phoP, and aroA were evaluated for their effects of deletion on safety and immunogenicity. Animal experiments demonstrated that C5001 (C500 rpoS+ Δcrp10) and C5002 (C500 rpoS+ Δfur9) showed an excellent ability to induce an immune response. To further decrease the endotoxic activity, the combination mutations of ΔpagL7 ΔpagP81::PlpplpxE ΔlpxR9 were introduced into the mutant strains to generate 1'-dephosphorylated lipid A. Animal experiments showed that SC3 (C500 rpoS+ Δfur9 ΔpagL7 ΔpagP81:: PlpplpxE ΔlpxR9) induced higher levels of IgG and secreted IgA antibodies and provided a higher protection rate than SC1 (C500 ΔpagL7 ΔpagP81:: PlpplpxE ΔlpxR9) and SC2 (C500 rpoS+ Δcrp10 ΔpagL7 ΔpagP81:: PlpplpxE ΔlpxR9). We also evaluated the ability of SC3 (C500 rpoS+ Δfur9 ΔpagL7 ΔpagP81:: PlpplpxE ΔlpxR9) as a vaccine carrier to deliver heterologous protein antigens and polysaccharide antigens. The results indicated that SC3 (C500 rpoS+ Δfur9 ΔpagL7 ΔpagP81:: PlpplpxE ΔlpxR9) showed an excellent ability to deliver heterologous antigens and induce the host to produce high levels of antibodies. Together, these results indicate that we constructed a safe and efficient attenuated strain of the S. Choleraesuis vaccine, which demonstrated strong resistance to infection with wild-type S. Choleraesuis and can be employed as a universal vector for the delivery of recombinant antigens.
[This corrects the article DOI: 10.3389/fpsyg.2023.1217290.].
Aeromonas hydrophila (A. hydrophila) is a typical zoonotic pathogenic bacterium that infects humans, animals, and fish. It has been reported that the Fur, a Fe2+ regulatory protein, and the Crp, a cAMP receptor protein, play important roles in bacterial virulence in many bacteria, but no research has been investigated on A. hydrophila. In this study, the Δfur and Δcrp mutant strains were constructed by the suicide plasmid method. These two mutant strains exhibited a slightly diminished bacterial growth and also were observed some alterations in the number of outer membrane proteins, and the disappearance of hemolysis in the Δcrp strain. Animal experiments of crucian carp showed that the Δfur and Δcrp mutant strains significantly decreased virulence compared to the wild-type strain, and both mutant strains were able to induce good immune responses by two kinds of administration routes of intraperitoneal immunization (i.p) and immersion immunization, and the protection rates through intraperitoneal injection of Δfur and Δcrp to crucian carp were as high as 83.3 % and 73.3 %, respectively, and immersion immunization route of Δfur and Δcrp to crucian carp provided protection as high as 40 % and 20 %, respectively. These two mutant strains showed abilities to induce changes in enzymatic activities of the non-specific enzymes SOD, LZM, AKP, and ACP in crucian carp. Together, these results indicated the Δfur and Δcrp mutants were safe and effective candidate vaccine strains, showing good protection against the wild-type A. hydrophila challenge.
Carps , Fish Diseases , Gram-Negative Bacterial Infections , Humans , Animals , Gram-Negative Bacterial Infections/prevention & control , Gram-Negative Bacterial Infections/veterinary , Vaccines, Attenuated , Aeromonas hydrophila
Objective: This study sought to uncover the relationship between physical education courses integrated with civic education (PECICE) and prosocial behavior among university students. Additionally, we aimed to decipher the mediating roles of cultural confidence and self-esteem within this relationship. Methods: Employing a questionnaire-based approach, we assessed the relationship among PECICE, cultural confidence, self-esteem, and prosocial behavior in university students. The instrument comprised four distinct scales: the Perceived Effectiveness Scale for PECICE, the Cultural Confidence Scale, the Self-Esteem Scale, and the Prosocial Behavior Scale. Our sample encompassed 293 Chinese college students, consisting of 137 men and 156 women, with an average age of 21.39 years (SD = 2.1). Results: PECICE demonstrated significant positive associations with cultural confidence (r = 0.29, p < 0.001), self-esteem (r = 0.35, p < 0.001), and prosocial behavior (r = 0.40, p < 0.001). The influence of PECICE on prosocial behavior among university students was mediated through three channels: solely via cultural confidence (mediating effect value: 0.14), solely via self-esteem (mediating effect value: 0.22), and through the combined influence of both cultural confidence and self-esteem (mediating effect value: 0.2). Conclusion: The intertwined mediating roles of cultural confidence and self-esteem highlight their pivotal significance in enhancing the efficacy of PECICE. These insights offer a valuable reference for both educators and policymakers striving to augment prosocial behavior in university students.
3D reconstruction is the process of obtaining the three-dimensional shape or surface structure of an object, which is widely used in advanced manufacturing fields such as automotive, aerospace, industrial inspection, and reverse engineering. However, due to the structural characteristics of the component itself, the reflective properties of the coating material, and other factors, there may be specular reflection during image acquisition, making it difficult to achieve complete 3D reconstruction of the component. This paper proposes a method to address the problem of incomplete 3D reconstruction of strongly reflective objects by recognizing outlier points and filling point cloud holes. The proposed View-Transform-PointNet outlier point recognition network improves the alignment of the initial point cloud plane and implements secondary alignment of the point cloud based on the perpendicularity between the outlier plane in mixed reflection and the point cloud plane. The point cloud hole-filling method is based on the principle of outlier formation and approximates a local Gaussian distribution to linear variation. The distance between the end of each outlier plane and the real surface is calculated to repair the depth information of outlier points. The proposed method achieves a 39.4% increase in the number of point cloud filling, a 45.2% increase in the number of triangular mesh faces, a 46.9% increase in surface area, and a chamfer distance (CD) of 0.4471009, which is better than existing geometric repair methods in terms of standard deviation and smoothness. The method improves the alignment of initial point cloud planes and enhances the accuracy of outlier point recognition, which are the main innovative points of this study. The 3D reconstruction of the repaired point cloud model is achieved through Poisson equation and parameter adjustment. The proposed method reduces the error caused by large curvature in the boundary region and improves the smoothness and accuracy of the reconstructed model.
Lipopolysaccharides (LPS) are the major constituent on the cell envelope of all gram-negative bacteria. They are ubiquitous in air, and are toxic inflammatory stimulators for urinary disorders and sepsis. The reported optical, thermal, and electrochemical sensors via the intermolecular interplay of LPS with proteins and aptamers are generally complicated methods. We demonstrate the single-molecule nanopore approach for LPS identification in distinct bacteria as well as the serotypes discrimination. With a 4 nm nanopore, we achieve a detection limit of 10 ng/mL. Both the antibiotic polymyxin B (PMB) and DNA aptamer display specific binding to LPS. The identification of LPS in both human serum and tap water show good performance with nanopore platforms. Our work shows a highly-sensitive and easy-to-handle scheme for clinical and environmental biomarkers determination and provides a promising screening tool for early warning of contamination in water and medical supplies.
Aptamers, Nucleotide , Biosensing Techniques , Nanopores , Humans , Lipopolysaccharides , Water
Aiming at the difficulty of traditional pesticide sampling, a low-cost and convenient flexible surface enhanced Raman scattering (SERS) gold core-silver shell-sponge (Au-Ag-sponge) substrate was synthesized by chemical reduction. The SERS substrate consisted of Au-AgNPs and a melamine sponge. The sponge had a rich open pore structure, which could well "capture" Au-AgNPs, generating a large number of "hot spots". The SERS enhancement activity of the flexible substrate was characterized with rhodamine 6G (R6G) Raman probe molecules. The substrate showed good activity to 10-12 M rhodamine 6G with an enhancement factor (EF) of 7.72 × 106. Applying this substrate to the qualitative and quantitative detection of pesticide residues, the results showed that the Raman intensity was well related to the concentration of pesticide solution with the range of 0.1-10 mg L-1 of thiram and 1-10 mg L-1 of diquat. Furthermore, the substrate was analyzed by finite difference time domain (FDTD) simulation and the results were in good agreement with the experimental results. The reason for the difference in Raman signals of pesticide molecules on the same substrate was the different binding modes of Au-AgNPs on the sponge. Finally, we pointed out the advantages of flexible substrates in the field of pesticide residues, as well as future opportunities and challenges.
Surface-enhanced Raman scattering (SERS) has a unique fingerprint spectrum, which allows for rapid, highly sensitive, and non-destructive detection without the need for sample pretreatment. However, SERS substrates have disadvantages such as short storage time and poor reproducibility. In this study, carbon nanotubes, gold, and silver were combined to take advantage of their inherent structural and characteristic properties that enhance the Raman effect. A new type of SERS composite substrate, CNTs/Ag@Au/SiO2, was prepared using a hydrothermal method and seed growth method. The substrate was characterized using transmission electron microscopy (TEM), and the average distance between the core-shell nanoparticles was found to be 3.1 nm, which is more suitable than other gold-silver combined core-shell structures and significantly improves the SERS enhancement factor. The substrate demonstrated high sensitivity even at low concentrations of probe molecules and good uniformity at five randomly selected locations. After storage for 45 days, the substrate still exhibited good stability. In most gold-silver combined core-shell structures, the detection limit for Rhodamine 6G (R6G) is 10-9 mol L-1, while in this substrate, the detection limit for R6G is 10-11 mol L-1. Furthermore, the contribution of the substrate's enhancement was deeply investigated using finite-difference time-domain (FDTD), which revealed that the substrate's hotspots were present in two forms: the "hotspots generated between Ag@Au nanoparticles" and the "hotspots generated between Ag@Au nanoparticles and carbon nanotubes". These two forms of hotspots also demonstrated that the performance brought about by the preparation of the substrate structure was reliable. The simulation results were compared with the experimental results, and the analysis showed that the real environment would have an impact on the substrate's structure during the actual substrate preparation process. Finally, the substrate was used for detecting the pesticide fipronil, and the results showed clear peaks even at a concentration of 0.1 mg L-1. The results indicated that the Raman intensity was linearly exponential with the fipronil solution concentration, with a determination coefficient of R2 = 0.991. This study provides a new SERS substrate for pesticide residue detection and further explores the improvement of pesticide detection limits.
Phenol red (PR) is generally used as an acid-base indicator and a printing and dyeing colorant. When its content exceeds a certain concentration in water, it will cause great damage to the human body. Therefore, it is very important to detect the content of PR in water. The advantage of surface enhanced Raman scattering (SERS) is detecting samples quickly, non-destructive and high sensitivity without sample pre-treatment. SERS has attracted great attention in all fields of detection and analysis. In this paper, the method of attaching silver nanoparticles to metallic single-walled carbon nanotubes form carbon nanotubes/silver nanoparticles (CNTs/AgNPs) structure and then combining it with silica sheet is proposed. SERS substrate with silica/carbon nanotubes/silver nanoparticles (SiO2/CNTs/AgNPs) composite structure has extremely high reinforcement effect. In the quantitative analysis of the detected substance, mathematical fitting or machine learning is used to find the relationship between the intensity of Raman signal and the concentration of the detected substance. The BP neural network optimized by genetic algorithm (GA-BP) is designed in this study. The weights of GA-BP to enhance the robustness of BP neural network, the method of adaptive learning rate and the number of hidden nodes is set to solve the problem that GA-BP is easy to fall into local optimum, thus establishing a quantitative analysis model of PR solution concentration. The model can detect different concentrations of PR solutions with high accuracy quickly, simply and sensitively. Finally, compared with other published quantitative models, GA-BP correlation coefficient R2 determined by the training results of the model is 0.99996, and the root mean square error of the prediction is RMSEP = 0.002510, which is 0.0005 higher than the mathematical fitting method, it shows better performance. A reliable idea for the preparation of SERS substrate and online detection of PR concentration in water proposed in this study.
The insecticide flonicamid (FLO) and its main degradation intermediate 4-trifluoromethylnicotinamide (TFNA-AM) are hazardous to the environment and animals. Microbial transformation of FLO has been well studied, but no study has yet reported on TFNA-AM degradation by a microorganism. Here, Pseudomonas stutzeri CGMCC 22915 effectively degraded TFNA-AM to 5-trifluoromethylnicotinic acid (TFNA). P. stutzeri CGMCC 22915 degraded 60.0% of TFNA-AM (1154.44 µmol/L) within 6 h with a half-life of just 4.5 h. Moreover, P. stutzeri CGMCC 22915 significantly promoted TFNA-AM decomposition in surface water. The reaction was catalyzed by an amidase, PsAmiA. PsAmiA is encoded in a novel nitrile-converting enzyme gene cluster. The enzyme shared only 20-44% identities with previously characterized signature amidases. PsAmiA was successfully expressed in Escherichia coli and its enzymatic properties were investigated using TFNA-AM as the substrate. PsAmiA was more active toward amides without hydrophilic groups, and did not hydrolyze another amide metabolite of FLO, N-(4-trifluoromethylnicotinoyl)glycinamide (TFNG-AM), which is structurally very similar to TFNA-AM. Molecular docking of PsAmiA and TFNA-AM indicated that hydrophobic residues Leu148, Ala150, Ala195, Ile225, Trp341, Leu460, and Ile463 may affect its substrate spectrum. This study provides new insights of the environmental fate of FLO at the molecular level and the structure-function relationships of amidases.
Insecticides , Amides , Amidohydrolases/genetics , Animals , Insecticides/chemistry , Molecular Docking Simulation , Niacinamide/analogs & derivatives , Nitriles , Water
Organic shell material and phase change material (PCM) have low thermal conductivity, which reduces the heat absorption and release rate of microencapsulated phase change materials (MEPCMs). Boron nitride nanosheets (BNNSs) with high thermal conductivity can not only stabilize the oil phase as the Pickering emulsifier but also improve the thermal conductivity of MEPCMs as one of the shell components, thus facilitating the heat conduction in the microcapsule system. Herein, MEPCM with paraffin wax (PW) as the core material and polystyrene (PS) modified by BNNSs as the shell material (PW@PS/BNNS MEPCMs) are synthesized via Pickering emulsion polymerization. The structure of PW@PS/BNNS MEPCMs can be regulated by tuning the PW and BNNS contents, to achieve high latent heat and thermal conductivity. In comparison to pure PW, the thermal conductivity of MEPCMs-5 wt % BNNSs increases by 63.76% at 25 °C. The PW@PS/BNNS powder possesses a latent heat capacity of 166.3 J/g, corresponding to a high encapsulation ratio of 80.77%. These properties endow the prepared MEPCMs with excellent thermal regulation properties. We also propose the formation mechanism of PW@PS/BNNS MEPCMs via Pickering emulsion polymerization for the first time, which will guide the MEPCM fabrication toward a reliable direction.
The three-dimensional trajectory tracking of AUV is an important basis for it to complete its task. Due to many uncertain disturbances such as wind, wave and current on the sea, it is easy to cause problems such as slow convergence speed of the controller and saturation of the controller output in the three-dimensional trajectory tracking control of AUV. And the dynamic uncertainty of AUV's own model will have a great negative impact on AUV's trajectory tracking control. In order to solve the problem of slow convergence speed of the above controller, the finite time control method is introduced into the designed position controller. In order to solve the problem of AUV controller output saturation, an auxiliary dynamic system is designed to compensate the system control output saturation. In order to solve the uncertainty of AUV model, a reduced order extended observer is designed in the dynamic controller. It can observe the motion parameters of AUV at any time, and compensate the uncertainty of model uncertainty and external environment disturbance in real time. The control method in this paper is simulated in a three-dimensional model. The experimental results show that the convergence speed, control accuracy, robustness and tracking effect of AUV are higher than those of common trajectory tracker. The algorithm is loaded into the "sea exploration â ¡" AUV and verified by experiments in Suzhou lake. The effect of AUV navigation basically meets the task requirements, in which the mean value of pitch angle and heading angle error is less than 8 degrees and the mean value of depth error is less than 0.1M. The trajectory tracker can better meet the trajectory tracking control needs of the AUV.
Subsurface damage of fused silica optics is one of the major factors restricting the performance of optical systems. The densification-affected deformation and fracture in fused silica under a sliding contact are investigated in this study, via three-dimensional finite element analysis (FEA). The finite element models of scratching with 70.3° conical and Berkovich indenters are established. A refined elliptical constitutive model is used to consider the influence of densification. The finite element models are experimentally verified by elastic recovery, and theoretically verified by hardness ratio. Results of densification and plastic deformation distributions indicate that the accuracy of existent sliding stress field models may be improved if the spherical/cylindrical yield region is replaced by an ellipsoid/cylindroid, and the embedding of the yield region is considered. The initiation sequence, and the locations and stages of radial, median, and lateral cracks are discussed by analyzing the predicted sliding stress fields. Median and radial cracks along the sliding direction tend to be the first cracks that emerge in the sliding and unloading stages, respectively. They coalesce to form a big median-radial crack that penetrates through the entire yield region. The fracture behavior of fused silica revealed in this study is essential in the low-damage machining of fused silica optics.
Colloidal molecules (CMs), nonspherical clusters of a small number of particles, can be used as building blocks for self-assembly applications. Here, we propose a novel one pot method for CMs synthesis. First, poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAM-co-AA)) microgels were prepared by soap-free emulsion polymerization as seed particles, then monomer styrene and cross-linking agent divinylbenzene (DVB) were added, which could be polymerized by the remaining free radicals on the seed surface in situ. P(NIPAM-co-AA)-PS colloidal molecules with a series of morphologies such as popcorn-like, CO2-like, NH3-like, CH4-like and so on could be obtained. The effects of satellite colloid viscosity, interfacial tension, and polymer chain mobility on the number of satellite colloid have been investigated, and the formation mechanism of CMs is proposed based on morphology evolution investigation. Compared with the existing CM synthesis techniques, our method enables fabricating CMs from vinyl monomer in a facile and efficient way, and the scientific finding regarding the CMs formation will guide the CMs fabrication toward salable and reliable direction.
PURPOSE: The conventional genetic screening for deafness involves 9-20 variants from four genes. This study expands screening to analyze the mutation types and frequency of hereditary deafness genes in Zhejiang, China, and explore the significance of in-depth deafness genetic screening in newborns. METHODS: This was a multi-centre study conducted in 5,120 newborns from 12 major hospitals in the East-West (including mountains and islands) of Zhejiang Province. Concurrent hearing and genetic screening was performed. For genetic testing, 159 variants of 22 genes were screened, including CDH23, COL11A1, DFNA5, DFNB59, DSPP, GJB2, GJB3, KCNJ10, MT-RNR1, MT-TL1, MT-TS1, MYO15A, MYO7A, OTOF, PCDH15, SLC26A4, SOX10, TCOF1, TMC1, USH1G, WFS1, and WHRN using next-generation sequencing. Newborns who failed to have genetic mutations or hearing screening were diagnosed audiologically at the age of 6 months. RESULTS: A total of 4,893 newborns (95.57%) have passed the initial hearing screening, and 7 (0.14%) have failed in repeated screening. Of these, 446 (8.71%) newborns carried at least one genetic deafness-associated variant. High-risk pathogenic variants were found in 11 newborns (0.21%) (nine homozygotes and two compound heterozygotes), and eight of these infants have passed the hearing screening. The frequency of mutations in GJB2, GJB3, SLC26A4, 12SrRNA, and TMC1 was 5.43%, 0.59%, 1.91%, 0.98%, and 0.02%, respectively. The positive rate of in-depth screening was significantly increased when compared with 20 variants in four genes of traditional testing, wherein GJB2 was increased by 97.2%, SLC26A4 by 21% and MT-RNR1 by 150%. The most common mutation variants were GJB2c.235delC and SLC26A4c.919-2A > G, followed by GJB2c.299_300delAT. Homoplasmic mutation in MT-RNR1 was the most common, including m.1555A > G, m.961T > C, m.1095T > C. All these infants have passed routine hearing screening. The positive rate of MT-RNR1 mutation was significantly higher in newborns with high-risk factors of maternal pregnancy. CONCLUSION: The positive rate of deafness gene mutations in the Zhejiang region is higher than that of the database, mainly in GJB2c.235delC, SLC26A4 c.919-2A > G, and m.1555A > G variants. The expanded genetic screening in the detection rate of diseasecausing variants was significantly improved. It is helpful in identifying high-risk children for follow-up intervention.
Despite ceaseless efforts in past decades, the memory effect of semi-crystalline polymers has not been elucidated completely yet. An important reason why is that residual lamellar crystals in the structured melt are difficult to characterize. Recently, we developed a new small-angle X-ray scattering (SAXS) theory [Li et al. (2019). IUCrJ, 6, 968-983] and Fourier transform method [Li et al. (2020). CrystEngComm, 22, 3042-3058] for lamellar crystals that could derive structural information from SAXS readily. In this study, we tried to employ the new theory and method to characterize residual lamellar crystals in the structured melt. It was found that although scattering peaks cannot be observed in raw scattering profiles, they actually exist. Subtracting free-melt scattering and multiplying by q 4 benefit the observation of these weak scattering peaks. With the new Fourier transform method, it was found that indeed as proposed previously, thicker lamellar crystals exist in the structured melt. To determine the lateral size of residual lamellar crystal especially, a new method was developed under the guidance of the new theory. With the new method, it was found that although the crystallinity is very low (â¼1% at 174°C), the lateral sizes in the structured melts are still large, e.g. 45.3â nm at 174°C, much greater than the critical nucleation size. This implies that these residual lamellar crystals can act as athermal nuclei after quenching to a lower temperature, as proposed by Ziabicki & Alfonso [(1994). Colloid Polym. Sci. 272, 1027-1042; (2002). Macromol. Symp. 185, 211-231] more than 20 years ago. The methodologies proposed here could also be applied to other polymer lamellar systems.
