Multimorbidity and mortality among older patients with coronary heart disease in Shenzhen,China

BACKGROUND The current understanding of the magnitude and consequences of multimorbidity in Chinese older adults with coronary heart disease(CHD)is insufficient.We aimed to assess the association and population-attributable fractions(PAFs)between multimorbidity and mortality among hospitalized older patients who were diagnosed with CHD in Shenzhen,China.METHODS We conducted a retrospective cohort study of older Chinese patients(aged≥65 years)who were diagnosed with CHD.Cox proportional hazards models were used to estimate the associations between multimorbidity and all-cause and cardiovascular disease(CVD)mortality.We also calculated the PAFs.RESULTS The study comprised 76,455 older hospitalized patients who were diagnosed with CHD between January 1,2016,and August 31,2022.Among them,70,217(91.9%)had multimorbidity,defined as the presence of at least one of the predefined 14 chronic conditions.Those with cancer,hemorrhagic stroke and chronic liver disease had the worst overall death risk,with adjusted HRs(95%CIs)of 4.05(3.77,4.38),2.22(1.94,2.53),and 1.85(1.63,2.11),respectively.For CVD mortality,the highest risk was observed for hemorrhagic stroke,ischemic stroke,and chronic kidney disease;the corresponding adjusted HRs(95%CIs)were 3.24(2.77,3.79),1.91(1.79,2.04),and 1.81(1.64,1.99),respectively.All-cause mortality was mostly attributable to cancer,heart failure and ischemic stroke,with PAFs of 11.8,10.2,and 9.1,respectively.As for CVD mortality,the leading PAFs were heart failure,ischemic stroke and diabetes;the corresponding PAFs were 18.0,15.7,and 6.1,respectively.CONCLUSIONS Multimorbidity was common and had a significant impact on mortality among older patients with CHD in Shenzhen,China.Cancer,heart failure,ischemic stroke and diabetes are the primary contributors to PAFs.Therefore,prioritizing improved treatment and management of these comorbidities is essential for the survival prognosis of CHD patients from a holistic public health perspective.

Importance of oxygen-containing functionalities and pore structures of biochar in catalyzing pyrolysis of homologous poplar

Biochar and bio-oil are produced simultaneously in one pyrolysis process,and they inevitably contact and may interact,influencing the composition of bio-oil and modifying the structure of biochar.In this sense,biochar is an inherent catalyst for pyrolysis.In this study,in order to investigate the influence of functionalities and pore structures of biochar on its capability for catalyzing the conversion of homologous volatiles in bio-oil,three char catalysts(600C,800C,and 800AC)produced via pyrolysis of poplar wood at 600 or 800℃or activated at 800℃,were used for catalyzing pyrolysis of homologous poplar wood at 600℃,respectively.The results indicated that the 600C catalyst was more active than 800C and 800AC for catalyzing cracking of volatiles to form more gas(yield increase by 40.2%)and aromatization of volatiles to form more light or heavy phenolics,due to its abundant oxygen-containing functionalities acting as active sites.The developed pores of the 800AC showed no such catalytic effect but could trap some volatiles and allow their further conversion via sufficient aromatization.Nevertheless,the interaction with the volatiles consumed oxygen on 600C(decrease by 50%),enhancing the aromatic degree and increasing thermal stability.The dominance of deposition of carbonaceous material of a very aromatic nature over 800C and 800AC resulted in net weight gain and blocked micropores but formed additional macropores.The in situ diffuse reflectance infrared Fourier transform spectroscopy characterization of the catalytic pyrolysis indicated superior activity of 600C for removal of -OH,while conversion of the intermediates bearing C=O was enhanced over all the char catalysts.

An effective strategy of constructing multi-metallic oxides of ZnO/ CoNiO_(2)/CoO/C microflowers for improved supercapacitive performance

In this work,a new ZnO/CoNiO_(2)/CoO/C metal oxides composite is prepared by cost-effective hydrothermal method coupled with annealing process under N_(2) atmosphere.Notably,the oxidation-defect annealing environment is conducive to both morphology and component of the composite,which flower-like ZnO/CoNiO_(2)/CoO/C is obtained.Benefited from good chemical stability of ZnO,high energy capacity of CoNiO_(2) and CoO and good conductivity of C,the as-prepared sample shows promising electrochemical behavior,including the specific capacity of 1435 C·g^(-1) at 1 A·g^(-1),capacity retention of 87.3%at 20 A·g^(-1),and cycling stability of 90.5%for 3000 cycles at 5 A·g^(-1),respectively.Furthermore,the prepared ZnO/CoNiO_(2)/CoO/C/NF//AC aqueous hybrid supercapacitors device delivers the best specific energy of 55.9 W·h·kg^(-1) at 850 W·kg^(-1).The results reflect that the as-prepared ZnO/CoNiO_(2)/CoO/C microflowers are considered as high performance electrode materials for supercapacitor,and the strategy mentioned in this paper is benefit to prepare mixed metal oxides composite for energy conversion and storage.

Advances of Synergistic Electrocatalysis Between Single Atoms and Nanoparticles/Clusters

Combining single atoms with clusters or nanoparticles is an emerging tactic to design efficient electrocatalysts.Both synergy effect and high atomic utilization of active sites in the composite catalysts result in enhanced electrocatalytic performance,simultaneously provide a radical analysis of the interrelationship between structure and activity.In this review,the recent advances of single-atomic site catalysts coupled with clusters or nanoparticles are emphasized.Firstly,the synthetic strategies,characterization,dynamics and types of single atoms coupled with clusters/nanoparticles are introduced,and then the key factors controlling the structure of the composite catalysts are discussed.Next,several clean energy catalytic reactions performed over the synergistic composite catalysts are illustrated.Eventually,the encountering challenges and recommendations for the future advancement of synergistic structure in energy-transformation electrocatalysis are outlined.

Design of a novel hybrid quantum deep neural network in INEQR images classification

We redesign the parameterized quantum circuit in the quantum deep neural network, construct a three-layer structure as the hidden layer, and then use classical optimization algorithms to train the parameterized quantum circuit, thereby propose a novel hybrid quantum deep neural network(HQDNN) used for image classification. After bilinear interpolation reduces the original image to a suitable size, an improved novel enhanced quantum representation(INEQR) is used to encode it into quantum states as the input of the HQDNN. Multi-layer parameterized quantum circuits are used as the main structure to implement feature extraction and classification. The output results of parameterized quantum circuits are converted into classical data through quantum measurements and then optimized on a classical computer. To verify the performance of the HQDNN, we conduct binary classification and three classification experiments on the MNIST(Modified National Institute of Standards and Technology) data set. In the first binary classification, the accuracy of 0 and 4 exceeds98%. Then we compare the performance of three classification with other algorithms, the results on two datasets show that the classification accuracy is higher than that of quantum deep neural network and general quantum convolutional neural network.

The role of a novel antibacterial substance,cyclic opine-producing Lacticaseibacillus rhamnosus LS8 in ameliorating ulcerative colitis:a fecal microbiota transplantation study

Intestinal microbiota imbalance may worsen the progression of ulcerative colitis(UC).Lacticaseibacillus rhamnosus LS8(LR)has the potential ability to regulate microbiota through producing a novel antibacterial substance,cyclic opine:cycloalanopine.This study aimed to investigate whether LR could ameliorate dextran sulfate sodium-induced UC in mice via modulating intestinal microbiota using fecal microbiota transplantation(FMT)experiment.The results showed that both LR and FMT attenuated UC as evidenced by 1)alleviating disease activity index and colonic pathology;2)up-regulating MUCs and tight junction proteins;3)increasing oxidative mediators and decreasing antioxidant mediators;4)down-regulating proinflammatory cytokines and chemokines.These results were mainly attributable to the microbiota-regulating effect of LR,including increasing beneficial bacteria(like Akkermansia)and its related SCFAs,while decreasing harmful bacteria(like Proteobacteria)and its related LPS,thereby suppressing the hyperactivation of TLR4/NF-κB pathway.Consequently,LR can alleviate UC and is a potential dietary supplement to attenuate UC.

Quantum generative adversarial networks based on a readout error mitigation method with fault tolerant mechanism

Readout errors caused by measurement noise are a significant source of errors in quantum circuits,which severely affect the output results and are an urgent problem to be solved in noisy-intermediate scale quantum(NISQ)computing.In this paper,we use the bit-flip averaging(BFA)method to mitigate frequent readout errors in quantum generative adversarial networks(QGAN)for image generation,which simplifies the response matrix structure by averaging the qubits for each random bit-flip in advance,successfully solving problems with high cost of measurement for traditional error mitigation methods.Our experiments were simulated in Qiskit using the handwritten digit image recognition dataset under the BFA-based method,the Kullback-Leibler(KL)divergence of the generated images converges to 0.04,0.05,and 0.1 for readout error probabilities of p=0.01,p=0.05,and p=0.1,respectively.Additionally,by evaluating the fidelity of the quantum states representing the images,we observe average fidelity values of 0.97,0.96,and 0.95 for the three readout error probabilities,respectively.These results demonstrate the robustness of the model in mitigating readout errors and provide a highly fault tolerant mechanism for image generation models.

Mitochondrial genomes of Tapes dorsatus and Cardita variegata:insights into Heteroconchia phylogeny

Heteroconchia,a widespread and abundant aquatic invertebrate,is an important clade of bivalve mollusks.The relationship between the three branches of Heteroconchia,Palaeoheterodonta,Archiheterodonta,and Euheterodonta has become a main controversy in molecular studies of the relationships between bivalves.In the present study,we assembled the complete mitochondrial genomes of Tapes dorsatus(Veneridae)and Cardita variegata(Carditidae)using high-throughput sequencing.C.variegata is the first mitochondrial genome belonging to the family Carditidae to be reported.We used 12 protein coding genes(excluding atp8)from the complete mitochondrial genomes of 146 species to recover the internal relationships of Heteroconchia.Our results support the traditional view of early branching of Palaeoheterodonta and the recovery of the monophyly of Palaeoheterodonta,Anomalodesmata,Imparidentia.Rearrangement analysis show that gene arrangement within Venerida was highly variable.Time-calibrated phylogenetic studies based on a relaxed molecular clock model suggested that Veneridae originated approximately 337.62 million years ago(Ma)and split into two major clades,whereas Carditidae originated approximately 510.09 Ma.Our results provide evidence of the internal relationships of Heteroconchia.

Ocular surface in patients with different degrees of myopia

AIM:To investigate the clinical features of the ocular surface in patients with different degrees of myopia.METHODS:A cross-sectional study was conducted involving 122 participants with myopia in Beijing Tongren Hospital from February to June,2023.After completing the Ocular Surface Disease Index(OSDI)score scale,measurements were taken for refraction,biometric parameters and ocular surface parameters.The prevalence,severity and related parameters of the dry eye among different groups based on axial length(AL)were compared.Correlation analysis was performed between ocular surface parameters and refraction/biometric measurement parameters.RESULTS:Statistically significant differences were observed in refractive error,corneal thickness,anterior chamber depth,and subfoveal choroidal thickness among the groups(all P<0.05).With the increase in AL,the incidence and severity of dry eye increased significantly(P<0.05).Moreover,the tear film break-up time(BUT)shortened(P<0.05),and the corneal fluorescein staining(CFS)points increased significantly(P<0.05).OSDI scores were positively correlated with AL and spherical equivalent(SE;both P<0.05);BUT was negatively correlated with AL,SE,and corneal astigmatism(AST;all P<0.05);Schirmer I test(SIT)results were negatively correlated with AL and SE(both P<0.05).CONCLUSION:AL elongation is a risk factor for dry eye onset in myopic participants.The longer the AL,the more severe the dry eye is,with the increased CFS spots and tear film instability.Additionally,SE and AST exhibit negative correlations with dry eye symptom scores and ocular surface parameters.

Analysis of learnability of a novel hybrid quantum-classical convolutional neural network in image classification

We design a new hybrid quantum-classical convolutional neural network(HQCCNN)model based on parameter quantum circuits.In this model,we use parameterized quantum circuits(PQCs)to redesign the convolutional layer in classical convolutional neural networks,forming a new quantum convolutional layer to achieve unitary transformation of quantum states,enabling the model to more accurately extract hidden information from images.At the same time,we combine the classical fully connected layer with PQCs to form a new hybrid quantum-classical fully connected layer to further improve the accuracy of classification.Finally,we use the MNIST dataset to test the potential of the HQCCNN.The results indicate that the HQCCNN has good performance in solving classification problems.In binary classification tasks,the classification accuracy of numbers 5 and 7 is as high as 99.71%.In multivariate classification,the accuracy rate also reaches 98.51%.Finally,we compare the performance of the HQCCNN with other models and find that the HQCCNN has better classification performance and convergence speed.

Gene characterization and phylogenetic analysis of four mitochondrial genomes in Caenogastropoda

Caenogastropoda is a highly diverse group,containing~60%of all existing gastropods.Species in this subclass predominantly inhabit marine environments and have a high ecological and economic value.Owing to the increase in relevant phylogenetic studies,our understanding of between species relatedness in Caenogastropoda has improved.However,the biodiversity,taxonomic status,and phylogenetic relationships of this group remain unclear.In the present study,we performed next-generation sequencing of four complete mitochondrial genomes from three families(Buccinidae,Columbellidae,and Cypraeidae)and the four mitogenomes were classical circular structures,with a length of 16177 bp in Volutharpa ampullacea,16244 bp in Mitrella albuginosa,16926bp in Mauritia arabica asiatica and 15422 bp in Erronea errones.Base composition analysis indicated that whole sequences were biased toward A and T.Then compared them with 171 complete mitochondrial genomes of Caenogastropoda.The phylogenetic relationship of Caenogastropoda derived from Maximum Likelihood(ML)and Bayesian Inference(BI)trees constructed based on CDS sequences was consistent with the results of traditional morphological analysis,with all three families showing close relationships.This study supported Caenogastropoda at the molecular level as a separate clade of Mollusca.According to our divergence time estimations,Caenogastropoda was formed during the middle Triassic period(~247.2–237 Ma).Our novel mitochondrial genomes provide evidence for the speciation of Caenogastropoda in addition to elucidating the mitochondrial genomic evolution of this subclass.

An Efficient Plant Regeneration System for Different Explants of Rare and Endangered Plants in Mussaenda anomala

To establish an efficient regeneration method for the rare and endangered plant Mussaenda anomala to address problems regarding its reproductive obstacles and scarce populations.In this study,the terminal buds,axillary buds,stem segments with two axillary buds,stem segments with two axillary buds and one terminal bud,and leaves of M.anomala were used as explants.The effects of different explants and disinfection methods,plant growth regulators and substrates on plant regeneration were explored.The following results were obtained:(1)The terminal bud was a suitable explant for M.anomala tissue culture,and the disinfection method utilized was treatment with 0.2%HgCl2 for 8 min.(2)Initiate medium:MS basic medium supplemented with 0.5 mg/L 6-BA and 0.2 mg/L IBA for the high germination rate(100%)and the maximum bud height(1.70 cm)of the terminal bud.(3)Proliferation medium:MS basic medium supplemented with 3.0 mg/L 6-BA and 0.2 mg/L IBA for a high proliferation rate(96%)and proliferation time(6.0)of terminal buds.(4)Proliferation medium supplemented with 0.7 mg/L GA3 significantly increased the bud heights of multiple buds.(5)Rooting medium:MS basic medium supplemented with 0.5 mg/L IBA and 0.5 mg/L IAA for a high rooting rate(88%),root number(12.0)and root length(5.07 cm).(6)The optimal substrate for seedling acclimation and transplanting was perlite:vermiculite(1:1),which resulted in the highest survival rate(97%)and plant height(5.89 cm),as well as better growth potential for seedlings.The surfaces of M.anomala explants are densely covered with trichome,which increased the difficulty of disinfection;the plant growth regulators directly affected the growth and development in the regeneration process of M.anomala,and the substrate significantly affected the survival rate and height growth for seedling acclimation.

肿瘤化疗患者焦虑和抑郁情绪与应对方式的关系研究

目的探讨肿瘤化疗患者的焦虑、抑郁情绪及不同应对方式对焦虑、抑郁的影响。方法选取2016年12月-2017年2月4所省、市级医院的339位肿瘤化疗患者为研究对象,分析研究肿瘤化疗患者的焦虑、抑郁状况与其应对方式的关系。结果 42.2%的化疗患者存在焦虑情绪,43.1%的化疗患者存在抑郁情绪,30.7%的患者两种情绪皆有。肿瘤化疗患者焦虑、抑郁得分均高于常模(P<0.05);面对得分低于国内常模(P<0.05),而回避和屈服得分均高于国内常模(P<0.05)。焦虑与屈服呈正相关(P<0.05);抑郁与面对呈负相关(P<0.05),与回避、屈服呈正相关(P<0.05)。多元逐步回归分析显示,医疗保障、职业及屈服是焦虑的影响因素(P<0.05);职业、屈服、面对及回避是焦虑的影响因素(P<0.05)。结论肿瘤化疗患者存在较为普遍的焦虑、抑郁情绪,这与其应对方式有一定的关系,尤其是屈服的应对方式对焦虑、抑郁情绪的产生具有重要影响。

滋肾化石汤辅助双镜联合术治疗复杂性肾结石的疗效及对红细胞免疫功能的影响

目的评价滋肾化石汤辅助双镜联合术治疗复杂性肾结石的疗效及对患者红细胞免疫功能的影响。方法该研究为前瞻性随机对照试验,选取2018年10月—2021年4月就诊于唐山市工人医院的108例复杂性肾结石患者。按随机数表法分组将患者分为对照组和实验组,每组54例。对照组接受经皮肾镜联合输尿管软镜碎石取石术,实验组实施经皮肾镜联合输尿管软镜碎石取石术+滋肾化石汤治疗。对比两组疗效、一期手术结石清除率、肾损伤因子、红细胞免疫功能、炎症应激反应、并发症。结果实验组总有效率、一期手术结石清除率较对照组高(P<0.05)。两组术前血清人肾损伤分子1(KIM-1)、中性粒细胞明胶酶相关载脂蛋白(NGAL)、尿素氮(BUN)及胱抑素C(Cys-C)水平比较,差异无统计学意义(P>0.05),实验组术后7 d血清KIM-1、NGAL、BUN及Cys-C水平较对照组低(P<0.05)。两组术前血清红细胞C3b受体花环率(C3bRR)、免疫复合物受体花环率(ICRRR)、肿瘤红细胞花环率(TER)及红细胞免疫复合物花环率(RBC-ICR)水平比较,差异无统计学意义(P>0.05),实验组术后7 d血清C3bRR、ICRRR、TER及RBC-ICR水平较对照组高(P<0.05)。两组术前血清白细胞介素-1β(IL-1β)、高敏C反应蛋白(hs-CRP)、血管紧张素Ⅰ(AngⅠ)及血管紧张素Ⅱ(AngⅡ)水平比较,差异无统计学意义(P>0.05)。实验组术后7 d血清血清IL-1β、hs-CRP、AngⅠ及AngⅡ水平较对照组低(P<0.05)。两组并发症发生率比较,差异无统计学意义(P>0.05)。结论复杂性肾结石患者接受滋肾化石汤辅助双镜联合术的效果确切,可提升一期手术结石清除率,减轻肾损伤,改善红细胞免疫功能,降低炎性应激反应。

超交联多孔材料的合成及吸附氨基酸特性

以苯为反应单体、二甲氧基甲烷(FDA)为交联剂,通过Friedel-Crafts烷基化反应,改变单体与交联剂之间的摩尔比来调控聚合物的孔隙结构,合成了一系列具有高比表面积的多孔芳香交联聚合物HCP-PhH-(1-3)。通过氮气吸附脱附、热失重和红外光谱对其进行表征,聚合物表现出丰富的孔径结构与良好的稳定性。将其对色氨酸(Trp)、苯丙氨酸(Phe)以及苯甘氨酸(Phg)进行了吸附行为研究,结果表明,单体苯与交联剂FDA的摩尔比为1:5时,合成的聚合物HCP-PhH-3具有相对较大的比表面积901.498 m^(2)/g,在25℃时对pH=7、浓度1.5 mmol/L的氨基酸溶液具有较大的吸附能力,吸附量分别为色氨酸(0.199 mmol/g)、苯丙氨酸(0.140mmol/g)、苯甘氨酸(0.034 mmol/g),显示出明显的吸附效果。对HCP-PhH-3吸附氨基酸的过程进行动力学拟合,线性拟合相关度(R2)符合准二级动力学模型。

含磷六元醇的制备及改性水性聚氨酯的结构与性能

以三氯氧磷、丙烯酸羟乙酯和二乙醇胺为原料,合成了一种含磷六羟基多元醇(NP6),将NP6作为内交联剂,异氟尔酮二异氰酸酯、聚丙二醇及二羟甲基丙酸等原料,合成了一系列NP6含量不同的聚氨酯乳液(WPU)。采用红外光谱、核磁共振对NP6和WPU的结构进行了表征,测试了乳液粒径、乳液稳定性和胶膜力学性能,采用热失重分析对乳胶膜的耐热性能进行了分析。结果表明,合成了与设计结构相符的NP6,制备的WPU具有良好的储存稳定性;乳液粒径随NP6含量增加先减小后增大,NP6的加入使乳胶膜的初始模量显著增加;加入NP6后乳胶膜的起始分解温度降低,分解后的残渣量增加。

微创玻璃体切除术中及术后脉络膜脱离的原因及治疗

无缝线微创玻璃体切除术的应用已有10余年,高速切割和精细器械的应用有效治疗严重的玻璃体视网膜疾病,但术中及术后的并发症也严重影响视力恢复,其中脉络膜脱离是微创玻璃体切除术中及术后比较少见的并发症,可继发青光眼、视力下降等。

Tooth regeneration: a revolution in stomatology and evolution in regenerative medicine

A tooth is a complex biological organ and consists of multiple tissues including the enamel, dentin, cementum and pulp. Tooth loss is the most common organ failure. Can a tooth be regenerated？ Can adult stem cells be orchestrated to regenerate tooth structures such as the enamel, dentin, cementum and dental pulp, or even an entire tooth？ If not, what are the therapeutically viable sources of stem cells for tooth regeneration？ Do stem cells necessarily need to be taken out of the body, and manipulated ex vivo before they are transplanted for tooth regeneration？ How can regenerated teeth be economically competitive with dental implants？ Would it be possible to make regenerated teeth affordable by a large segment of the population worldwide？ This review article explores existing and visionary approaches that address some of the above-mentioned questions. Tooth regeneration represents a revolution in stomatology as a shift in the paradigm from repair to regeneration： repair is by metal or artificial materials whereas regeneration is by biological restoration. Tooth regeneration is an extension of the concepts in the broad field of regenerative medicine to restore a tissue defect to its original form and function by biological substitutes.

Histogram analysis of apparent diffusion coefficient predicts response to radiofrequency ablation in hepatocellular carcinoma

Objective: The aim of this study was to predict tumor progression in patients with hepatocellular carcinoma(HCC) treated with radiofrequency ablation(RFA) using histogram analysis of apparent diffusion coefficients(ADC).Methods: Breath-hold diffusion weighted imaging(DWI) was performed in 64 patients(33 progressive and 31 stable) with biopsy-proven HCC prior to RFA. All patients had pre-treatment magnetic resonance imaging(MRI)and follow-up computed tomography(CT) or MRI. The ADC values(ADC_(10), ADC_(30_, ADC_(median) and ADC_(max))were obtained from the histogram's 10 th, 30 th, 50 th and 100 th percentiles. The ratios of ADC_(10), ADC_(30_,ADCmedian and ADCmax to the mean non-lesion area-ADC(RADC_(10), RADC_(30_, RADC_(median), and RADC_(max)) were calculated. The two patient groups were compared. Key predictive factors for survival were determined using the univariate and multivariate analysis of the Cox model. The Kaplan-Meier survival analysis was performed, and pairs of survival curves based on the key factors were compared using the log-rank test.Results: The ADC_(30_, ADCmedian, ADCmax, RADC_(30_, RADC_(median), and RADC_(max) were significantly larger in the progressive group than in the stable group(P<0.05). The median progression-free survival(PFS) was 22.9 months for all patients. The mean PFS for the stable and progressive groups were 47.7±1.3 and 9.8±1.3 months,respectively. Univariate analysis indicated that RADC_(10), RADC_(30_, and RADC_(median) were significantly correlated with the PFS [hazard ratio(HR)=31.02, 43.84, and 44.29, respectively, P<0.05 for all]. Multivariate analysis showed that RADCmedian was the only independent predictor of tumor progression(P=0.04). And the cutoff value of RADC_(median) was 0.71.Conclusions: Pre-RFA ADC histogram analysis might serve as a useful biomarker for predicting tumor progression and survival in patients with HCC treated with RFA.

Development and in vitro study of a bi-specific magnetic resonance imaging molecular probe for hepatocellular carcinoma

BACKGROUND Hepatocellular carcinoma(HCC)ranks second in terms of cancer mortality worldwide.Molecular magnetic resonance imaging(MRI)targeting HCC biomarkers such as alpha-fetoprotein(AFP)or glypican-3(GPC3)offers new strategies to enhance specificity and help early diagnosis of HCC.However,the existing iron oxide nanoparticle-based MR molecular probes singly target AFP or GPC3,which may hinder their efficiency to detect heterogeneous micro malignant HCC tumors<1 cm(MHCC).We hypothesized that the strategy of double antibody-conjugated iron oxide nanoparticles which simultaneously target AFP and GPC3 antigens may potentially be used to overcome the tumor heterogeneity and enhance the detection rate for MRI-based MHCC diagnosis.AIM To synthesize an AFP/GPC3 double antibody-labeled iron oxide MRI molecular probe and to assess its impact on MRI specificity and sensitivity at the cellular level.METHODS A double antigen-targeted MRI probe for MHCC anti-AFP-USPIO-anti-GPC3(UAG)was developed by simultaneously conjugating AFP andGPC3 antibodies to a 5 nm ultra-small superparamagnetic iron oxide nanoparticle(USPIO).At the same time,the singly labeled probes of anti-AFP-USPIO(UA)and anti-GPC3-USPIO(UG)and non-targeted USPIO(U)were also prepared for comparison.The physical characterization including morphology(transmission electron microscopy),hydrodynamic size,and zeta potential(dynamic light scattering)was conducted for each of the probes.The antigen targeting and MRI ability for these four kinds of USPIO probes were studied in the GPC3-expressing murine hepatoma cell line Hepa1-6/GPC3.First,AFP and GPC3 antigen expression in Hepa1-6/GPC3 cells was confirmed by flow cytometry and immunocytochemistry.Then,the cellular uptake of USPIO probes was investigated by Prussian blue staining assay and in vitro MRI(T2-weighted and T2-map)with a 3.0 Tesla clinical MR scanner.RESULTS Our data showed that the double antibody-conjugated probe UAG had the best specificity in targeting Hepa1-6/GPC3 cells expressing AFP and GPC3 antigens compared with single antibody-conjugated and unconjugated USPIO probes.The iron Prussian blue staining and quantitative T2-map MRI analysis showed that,compared with UA,UG,and U,the uptake of double antigen-targeted UAG probe demonstrated a 23.3%(vs UA),15.4%(vs UG),and 57.3%(vs U)increased Prussian stained cell percentage and a 14.93%(vs UA),9.38%(vs UG),and 15.3%(vs U)reduction of T2 relaxation time,respectively.Such bi-specific probe might have the potential to overcome tumor heterogeneity.Meanwhile,the coupling of two antibodies did not influence the magnetic performance of USPIO,and the relatively small hydrodynamic size(59.60±1.87 nm)of double antibodyconjugated USPIO probe makes it a viable candidate for use in MHCC MRI in vivo,as they are slowly phagocytosed by macrophages.CONCLUSION The bi-specific probe presents enhanced targeting efficiency and MRI sensitivity to HCC cells than singly-or non-targeted USPIO,paving the way for in vivo translation to further evaluate its clinical potential.