Macromolecular Nano-Assemblies for Enhancing the Effect of Oxygen-Dependent Photodynamic Therapy Against Hypoxic Tumors.

In oxygen (O2)-dependent photodynamic therapy (PDT), photosensitizers absorb light energy, which is then transferred to ambient O2 and subsequently generates cytotoxic singlet oxygen (1O2). Therefore, the availability of O2 and the utilization efficiency of generated 1O2 are two significant factors that influence the effectiveness of PDT. However, tumor microenvironments (TMEs) characterized by hypoxia and limited utilization efficiency of 1O2 resulting from its short half-life and short diffusion distance significantly restrict the applicability of PDT for hypoxic tumors. To address these challenges, numerous macromolecular nano-assemblies (MNAs) have been designed to relieve hypoxia, utilize hypoxia or enhance the utilization efficiency of 1O2. Herein, we provide a comprehensive review on recent advancements achieved with MNAs in enhancing the effectiveness of O2-dependent PDT against hypoxic tumors.

Molecular fingerprints of nuclear genome and mitochondrial genome for early diagnosis of lung adenocarcinoma.

BACKGROUND: Lung adenocarcinoma (LUAD) is the most prevalent subtype of lung cancer with high morbidity and mortality rates. Due to the heterogeneity of LUAD, its characteristics remain poorly understood. Exploring the clinical and molecular characteristics of LUAD is challenging but vital for early diagnosis. METHODS: This observational and validation study enrolled 80 patients and 13 healthy controls. Nuclear and mtDNA-captured sequencings were performed. RESULTS: This study identified a spectrum of nuclear and mitochondrial genome mutations in early-stage lung adenocarcinoma and explored their association with diagnosis. The correlation coefficient for somatic mutations in cfDNA and patient-matched tumor tissues was high in nuclear and mitochondrial genomes. The mutation number of highly mutated genes was evaluated, and the Least Absolute Shrinkage and Selection Operator (LASSO) established a diagnostic model. Receiver operating characteristic (ROC) curve analysis explored the diagnostic ability of the two panels. All models were verified in the testing cohort, and the mtDNA panel demonstrated excellent performance. This study identified somatic mutations in the nuclear and mitochondrial genomes, and detecting mutations in cfDNA displayed good diagnostic performance for early-stage LUAD. Moreover, detecting somatic mutations in the mitochondria may be a better tool for diagnosing early-stage LUAD. CONCLUSIONS: This study identified specific and sensitive diagnostic biomarkers for early-stage LUAD by focusing on nuclear and mitochondrial genome mutations. This also further developed an early-stage LUAD-specific mutation gene panel for clinical utility. This study established a foundation for further investigation of LUAD molecular pathogenesis.

Electrochemical behaviors of Li-B alloys in a LiCl-LiBr-KBr molten salt system.

Li-B alloys present higher voltages and better power performances than those of conventional Li-Al and Li-Si anodes for thermal batteries. Herein, the electrochemical characteristics of the Li-B alloy in the LiCl-LiBr-KBr electrolyte, including the discharge mechanism, charge transfer coefficient and exchange current density, were investigated in the temperature range of 623-823 K by open circuit potential (OCP), cyclic voltammetry (CV), chronopotentiometry (CP), linear sweep voltammetry (LSV) and electrochemical impedance spectroscopy (EIS) techniques. Consequently, the OCP of the Li-B alloy in the LiCl-LiBr-KBr electrolyte is close to that of pure lithium at the investigated temperatures. The discharge of the Li-B alloy electrode includes electrochemical dissolution of free lithium (Li → Li+) and compounded lithium (LiB → Li+ + B). The charge transfer coefficient in the anodic direction (Li → Li+) is about 0.63 at 623 K, which slightly increases as the temperature increases. The exchange current density of the Li (Li-B)/Li+ couple determined by the EIS method increases from 3.84 A cm-2 to 8.40 A cm-2 when the temperature increases from 623 to 823 K, corresponding to an activation energy of 16.4 kJ mol-1. These results suggest that the Li-B anode allows ultrahigh-rate discharge in thermal batteries.

Codon usage bias regulates gene expression and protein conformation in yeast expression system P. pastoris.

BACKGROUND: Protein synthesis is one of the extremely important anabolic pathways in the yeast expression system Pichia pastoris. Codon optimization is a commonly adopted strategy for improved protein expression, although unexpected failures did appear sometimes waiting for further exploration. Recently codon bias has been studied to regulate protein folding and activity in many other organisms. RESULTS: Here the codon bias profile of P. pastoris genome was examined first and a direct correlation between codon translation efficiency and usage frequency was identified. By manipulating the codon choices of both endogenous and heterologous signal peptides, secretion abilities of N-terminal signal peptides were shown to be tolerant towards codon changes. Then two gene candidates with different levels of structural disorder were studied, and full-length codon optimization was found to affect their expression profiles differentially. Finally, more evidences were provided to support possible protein conformation change brought by codon optimization in structurally disordered proteins. CONCLUSION: Our results suggest that codon bias regulates gene expression by modulating several factors including transcription and translation efficiency, protein folding and activity. Because of sequences difference, the extent of affection may be gene specific. For some genes, special codon optimization strategy should be adopted to ensure appropriate expression and conformation.

Dual-probe fluorescent biosensor based on T7 exonuclease-assisted target recycling amplification for simultaneous sensitive detection of microRNA-21 and microRNA-155.

Effective and simultaneous monitoring of the abnormal expression of certain microRNAs (miRNAs), especially for miRNA-21 and miRNA-155, can indicate drug resistance in lung cancer. In this work, T7 exonuclease (T7 Exo)-assisted target recycling amplification coupled with the extensive fluorescence quenching of graphene oxide (GO) was designed for the simultaneous detection of miRNA-21 and miRNA-155 using FAM- and ROX-labeled single-strand DNA probes. Through this method, the variable emission intensities of FAM and ROX caused by the introduction of miRNA-21 and miRNA-155, respectively, were obtained with high sensitivity. The method exhibited excellent analytical performance for simultaneous detection of miRNA-21 and miRNA-155 without cross-interference. The linear range was from 0.005 nM to 5 nM over three orders of magnitude, with detection limits as low as 3.2 pM and 4.5 pM for miRNA-21 and miRNA-155, respectively. Furthermore, the recovery (92.49-103.67%) and relative standard deviation (RSD < 4.8%) of the standard addition test of miRNA-21 and miRNA-155 in human plasma suggested the potential for drug resistance warning in clinical practice via this simple strategy. A homogeneous T7 Exo-assisted signal amplification combined with GO quenching platform was developed for accurate, sensitive and simultaneous analysis of miRNA-21 and miRNA-155 for drug resistance warning in lung cancer. This simple method exhibited a wide linear range and low LODs for miR-21 and miR-155.

Codon usage bias affects α-amylase mRNA level by altering RNA stability and cytosine methylation patterns in Escherichia coli.

Codon usage bias exists in almost every organism and is reported to regulate protein translation efficiency and folding. Besides translation, the preliminary role of codon usage bias on gene transcription has also been revealed in some eukaryotes such as Neurospora crassa. In this study, we took as an example the α-amylase-coding gene (amyA) and examined the role of codon usage bias in regulating gene expression in the typical prokaryote Escherichia coli. We confirmed the higher translation efficiency on codon-optimized amyA RNAs and found that the RNA level itself was also affected by codon optimization. The decreased RNA level was caused at least in part by altered mRNA stability at the post-transcriptional level. Codon optimization also altered the number of cytosine methylation sites. Examination on dcm knockouts suggested that cytosine methylation may be a minor mechanism adopted by codon bias to regulate gene RNA levels. More studies are required to verify the global effect of codon usage and to reveal its detailed mechanism on transcription.

A dual-label time-resolved fluorescence immunoassay for screening of osteoporosis based on simultaneous detection of C-terminal telopeptide (ß-CTX) and aminoterminal propeptide (P1NP) of type I procollagen.

Osteoporosis is a disease where increased bone weakness increases the risk of a broken bone. Until a broken bone occurs, there are typically no symptoms. Osteoporosis affects more than 75 million people in the United States, Europe and Japan. The diagnosis of osteoporosis is primarily determined by measuring bone mineral density using dual-energy X-ray absorptiometry, but for men under 50 years of age, premenopausal women should not be made on the basis of densitometric criteria alone. Bone biomarkers are a useful tool in detecting osteoporotic. A two-step dual-label time-resolved fluorescence immunoassay (TRFIA) was developed for the simultaneous detection of serum C-terminal telopeptide (ß-CTX) and amino-terminal propeptide (P1NP) of Type I procollagen in a single run. The performance of this assay was first evaluated using clinical serum samples, and then compared with commercialized kits. The sensitivity of this assay for ß-CTX was 1 ng/L (dynamic range, 0-1000 ng/L), and the sensitivity for P1NP detection was 1 µg/L (dynamic range, 1-1000 µg/L). High correlation coefficients (R) were obtained between the present dual-label TRFIA and commercially available kits (R = 0.99 for ß-CTX and P1NP). The present dual-label TRFIA has high sensitivity, specificity and accuracy in clinical sample analysis. It is a good alternative to the single-label diagnostic methods.

A new method to predict the outcome of the volar locked plate treatment for distal radius fracture.

BACKGROUND: Volar locked plate for distal radius fracture is one of the common procedures performed in trauma surgery. There are already some factors which can be used to predict the functional outcome after volar locked plating for distal radius fracture. However their limitations caused that the outcomes couldn't be satisfactorily predicted. Better factors for predicting the prognosis more precisely are of great interest. The aim of this study is to introduce such a new factor. METHODS: A total of 56 patients suffered from unilateral distal radius fracture were managed operatively with the volar locked plate. Before operation, all CT scans of the distal radius were obtained. The ratios of soft tissue circumference to bone circumference at the watershed line in the distal radius were calculated based on the preoperative CT scans. Outcomes were evaluated after operation. The correlations between the ratio and the outcomes were analyzed using single factor linear regression analysis. RESULTS: Statistically significant linear relationships between the ratio and flexion degrees, extension degrees also patient-rated wrist evaluation (PRWE) scores were discovered. With the increase of the ratios, the flexion and extension range increased and the PRWE scores declined. CONCLUSIONS: There are obvious linear relationships between the ratio and postoperative wrist flexion-extension degrees also PRWE scores when using volar locked plating for distal radius fracture. So the ratio can be used as a predictor aiding surgeons to predict the outcome.

Aquaporin-3 Attenuates Oxidative Stress-Induced Nucleus Pulposus Cell Apoptosis Through Regulating the P38 MAPK Pathway.

BACKGROUND/AIMS: Previous studies have shown that oxidative damage is a main contributor to disc nucleus pulposus (NP) cell apoptosis. Aquaporin-3 (AQP-3) facilitates reactive oxygen species (ROS) scavenging and thus alleviates oxidative injury in other cells. This study aims to investigate the role and mechanism of AQP-3 in regulating NP cell apoptosis under oxidative damage. METHODS: Rat NP cells were treated with H2O2 for 48 hours, while control NP cells were free of H2O2. Recombinant AQP-3 lentiviral vectors were used to investigate the effect of enhanced AQP-3 expression levels in NP cells. NP cell apoptosis was assessed by flow cytometry, caspase-3 activity, gene expression of apoptosis-related molecules (Bax, Bcl-2 and caspase-3), and protein expression of cellular apoptosis markers (cleaved PARP and cleaved caspase-3). Additionally, intracellular ROS content and activity of the p38 MAPK pathway were evaluated. RESULTS: Compared with the control NP cells, oxidative damage in the treatment cells significantly increased cell apoptosis ratios and caspase-3 activity, upregulated gene expression of Bax and caspase-3, downregulated gene expression of Bcl-2, and increased protein expression of cleaved PARP and cleaved caspase-3, as well as increased intracellular ROS content and activity of the p38 MAPK pathway. However, AQP-3 overexpression partly alleviated cell apoptosis, decreased intracellular ROS content, and inhibited the p38 MAPK pathway in NP cells under oxidative damage. CONCLUSION: Oxidative damage can significantly downregulate AQP-3 expression. Enhancing AQP-3 expression in NP cells partly attenuates cellular apoptosis through regulating the p38 MAPK pathway under oxidative damage.

Dynamic Compression Promotes the Matrix Synthesis of Nucleus Pulposus Cells Through Up-Regulating N-CDH Expression in a Perfusion Bioreactor Culture.

BACKGROUND/AIMS: An adequate matrix production of nucleus pulposus (NP) cells is an important tissue engineering-based strategy to regenerate degenerative discs. Here, we mainly aimed to investigate the effects and mechanism of mechanical compression (i.e., static compression vs. dynamic compression) on the matrix synthesis of three-dimensional (3D) cultured NP cells in vitro. METHODS: Rat NP cells seeded on small intestinal submucosa (SIS) cryogel scaffolds were cultured in the chambers of a self-developed, mechanically active bioreactor for 10 days. Meanwhile, the NP cells were subjected to compression (static compression or dynamic compression at a 10% scaffold deformation) for 6 hours once per day. Unloaded NP cells were used as controls. The cellular phenotype and matrix biosynthesis of NP cells were investigated by real-time PCR and Western blotting assays. Lentivirus-mediated N-cadherin (N-CDH) knockdown and an inhibitor, LY294002, were used to further investigate the role of N-CDH and the PI3K/Akt pathway in this process. RESULTS: Dynamic compression better maintained the expression of cell-specific markers (keratin-19, FOXF1 and PAX1) and matrix macromolecules (aggrecan and collagen II), as well as N-CDH expression and the activity of the PI3K/Akt pathway, in the 3D-cultured NP cells compared with those expression levels and activity in the cells grown under static compression. Further analysis showed that the N-CDH knockdown significantly down-regulated the expression of NP cell-specific markers and matrix macromolecules and inhibited the activation of the PI3K/Akt pathway under dynamic compression. However, inhibition of the PI3K/Akt pathway had no effects on N-CDH expression but down-regulated the expression of NP cell-specific markers and matrix macromolecules under dynamic compression. CONCLUSION: Dynamic compression increases the matrix synthesis of 3D-cultured NP cells compared with that of the cells under static compression, and the N-CDH-PI3K/Akt pathway is involved in this regulatory process. This study provides a promising strategy to promote the matrix deposition of tissue-engineered NP tissue in vitro prior to clinical transplantation.

N-Cadherin Attenuates High Glucose-Induced Nucleus Pulposus Cell Senescence Through Regulation of the ROS/NF-κB Pathway.

BACKGROUND/AIMS: Diabetes mellitus (DM) is a potential etiology of disc degeneration. N-cadherin (N-CDH) helps maintain the cell viability, cell phenotype and matrix biosynthesis of nucleus pulposus (NP) cells. Here, we mainly aimed to investigate whether N-CDH can attenuate high glucose-induced NP cell senescence and its potential mechanism. METHODS: Rat NP cells were cultured in a base culture medium and base culture medium with a 0.2 M glucose concentration. Recombinant lentiviral vectors were used to enhance N-CDH expression in NP cells. Senescence-associated ß-galactosidase (SA-ß-Gal) activity was measured by SA-ß-Gal staining. NP cell proliferation was evaluated by CCK-8 assay. Telomerase activity and intracellular reactive oxygen species (ROS) content were tested by specific chemical kits according to the manufacturer's instructions. G0/G1 cell cycle arrest was evaluated by flow cytometry. Real-time PCR and Western blotting were used to analyze mRNA and protein expressions of senescence markers (p16 and p53) and matrix macromolecules (aggrecan and collagen II). Additionally, p-NF-κB expression was also analyzed by Western blotting to evaluate NF-κB pathway activity. RESULTS: High glucose significantly decreased N-CDH expression, increased ROS generation and NF-κB pathway activity, and promoted NP cell senescence, which was reflected in the increase in SA-ß-Gal activity and senescence marker (p16 and p53) expression, compared to the control group. High glucose decreased telomerase activity and cell proliferation potency. However, N-CDH overexpression partially attenuated NP cell senescence, decreased ROS content and inhibited the activation of the NF-κB pathway under the high glucose condition. CONCLUSION: High glucose decreases N-CDH expression and promotes NP cell senescence. N-CDH overexpression can attenuate high glucose-induced NP cell senescence through the regulation of the ROS/ NF-κB pathway. This study suggests that N-CDH is a potential therapeutic target to slow DM-mediated disc NP degeneration.

N-Cadherin-Mediated Activation of PI3K/Akt-GSK-3ß Signaling Attenuates Nucleus Pulposus Cell Apoptosis Under High-Magnitude Compression.

BACKGROUND/AIMS: Mechanical overloading-induced nucleus pulposus (NP) apoptosis plays an important role in the pathogenesis of intervertebral disc degeneration. N-cadherin (N-CDH)-mediated signaling preserves normal NP cell phenotype. This study aims to investigate the effects of N-CDH on NP cell apoptosis under high-magnitude compression and the underlying mechanism behind this process. METHODS: Rat NP cells seeded on scaffold were perfusion-cultured using a self-developed perfusion bioreactor for 5 days and experienced different magnitudes (2% and 20% compressive deformation, respectively) of compression at a frequency of 1.0 Hz for 4 hours once per day. The un-loaded NP cells were used as controls. Lentivirus-mediated N-CDH overexpression and inhibitor LY294002 were used to further investigate the role of N-CDH and PI3K/Akt pathway under high-magnitude compression, respectively. NP cell apoptosis was evaluated by caspase-3 activity measured using a commercial kit, flow cytometry, and expression of apoptosis-related molecules analyzed by real-time PCR and western blotting assays. RESULTS: High-magnitude compression significantly increased apoptotic NP cells, caspase-3 activity and expression of pro-apoptotic molecules (Bax and caspase-3/cleaved caspase-3), but decreased expression of anti-apoptotic molecule (Bcl-2). High-magnitude compression decreased expression of N-CDH, p-Akt and p-GSK-3ß. However, N-CDH overexpression attenuated NP cell apoptosis and increased expression of p-Akt and p-GSK-3ß under high-magnitude compression. Further analysis showed that inhibition of the PI3K/Akt pathway suppressed NP cell apoptosis and decreased expression of p-GSK-3ß, but had no significant effects on N-CDH expression under high-magnitude compression. CONCLUSION: N-CDH can attenuate NP cell apoptosis through activating the PI3K/Akt-GSK-3ß signaling under high-magnitude compression.

Lnc-ATB contributes to gastric cancer growth through a MiR-141-3p/TGFß2 feedback loop.

The long noncoding RNA (lncRNA) ATB is an important regulator in human tumors. Here, we aimed to investigate the potential molecular mechanisms of lnc-ATB in gastric cancer (GC) tumorigenesis. RT-qPCR analysis was used to detect lnc-ATB expression level in 20 pairs of gastric cancer tissues and adjacent normal gastric mucosa tissues (ANTs). Moreover, the biological role of lnc-ATB was determined in vitro. We found that lnc-ATB was significantly upregulated in GC tissues compared to lnc-ATB expression in ANTs. These high lnc-ATB expression levels predicted poor prognosis in GC patients. Low levels of lnc-ATB inhibited GC cell proliferation and cell cycle arrest in vitro. Lnc-ATB was found to directly bind miR-141-3p. Moreover, TGF-ß actives lnc-ATB and TGF-ß2 directly binds mir-141-3p. Finally, we demonstrated that lnc-ATB fulfilled its oncogenic roles in a ceRNA-mediated manner. Our study suggests that lnc-ATB promotes tumor progression by interacting with miR-141-3p and that Lnc-ATB may be a valuable prognostic predictor for GC. In conclusion, the positive feedback loop of lnc-ATB/miR-141-3p/TGF-ß2 may be a potential therapeutic target for the treatment of GC.

The influence of liquid Pb-Bi on the anti-corrosion behavior of Fe3O4: a first-principles study.

In this work, the influence of Pb and Bi atoms on the anti-corrosion behavior of the oxide film (Fe3O4) formed on steel surface is investigated based on first-principles calculations. Through calculations of the formation energies, we find that Pb and Bi atoms can promote the formation of point defects, such as interstitial atoms and vacancies in Fe3O4. Besides, the effects of the concentration of Pb (or Bi) and pressure on the formation of these defects are also studied. Our results depict that a high density of Pb (or Bi) and compression pressure can promote the formation of defects in Fe3O4 significantly. Furthermore, the energy barriers for Pb and Bi atom migration in Fe3O4 are also estimated using the climbing image nudge elastic band (CI-NEB) method, which implies that Pb and Bi can diffuse more easily in Fe3O4 compared to Fe. Our results reveal the underlying mechanism of how Pb and Bi influence the anti-corrosion ability of oxide films in an accelerate driven system (ADS). It is instructive for improving the corrosion resistance of the oxide films in the ADS.

3B, a novel of photosensitizer, exhibited anti-tumor effects via mitochondrial apoptosis pathway in MCF-7 human breast carcinoma cells.

Photodynamic therapy (PDT) has been recognized as an innovated therapeutic modality for the treatment of various cancers. In this study, we evaluated the anticancer effect of a new photosensitizer 3B in breast cancer, which was considered one of the most common cancers in women worldwide. Here, we determined the effect of 3B not only on the cell growth, apoptosis, and Bcl-2 signal pathway in vitro but also on the anti-cancer effect in nude mice in vivo. Our results showed that 3B was primarily accumulated in mitochondria, increased the level of ROS, induced apoptotic cells death via Bcl-2 family, and its activity could be blocked by the caspase inhibitor (Z-VAD-FMK). In vivo study, 3B made a significant opening inhibition of tumor growth and showed drug toxicity hardly. TUNEL assay indicated that PDT group showed more positive cells (green) than other groups. These data supported that 3B might develop as potential therapeutic drug for the treatment of breast cancer.

Effect of irradiation defects on the corrosion behaviors of steels exposed to lead bismuth eutectic in ADS: a first-principles study.

In accelerator driven systems (ADSs), steels will suffer not only from the irradiation damage produced by protons or neutrons, but also from the dissolution corrosion induced by the liquid lead-bismuth eutectic (LBE). In this work we investigate the interactions between LBE atoms (Pb, Bi) and the irradiation induced defects X (X is helium, vacancy or divacancy) in α-Fe based on first-principles calculations. It is found that LBE atoms repulse each other without irradiation defects, while they aggregate easily with the defects to form X-Pbn and X-Bin complexes. This indicates that the irradiation defects could promote the aggregation of LBE atoms in iron, especially Bi atoms. The total binding energies of the X-Pbn and X-Bin complexes increase with the number of Pb and Bi atoms, respectively. The origin of the total binding energies of the complexes is further discussed via the electronic structures and the distortion of the crystalline lattice. Finally, the concentration evolutions of the Vac-(Bi)n complexes and unbound vacancies with temperature are predicted by the mass action analysis. This work provides important information for the synergistic effect of irradiation and LBE corrosion on the steels in the ADSs, which can be used as basic parameters for further study.

Comparison of multiplex immunochemical and molecular serotyping methods for Shiga toxin-producing Escherichia coli.

Traditionally, serotyping of Escherichia coli has been performed via slide agglutination methods using antisera. More recently, multiplex immunoassays and "molecular serotyping" via polymerase chain reaction (PCR) have been validated for this purpose. In this study, the serogroups of 161 Shiga toxin-producing Escherichia coli (STEC) strains isolated from fecal samples of California cattle were typed by conventional methods using antisera as well as two newly developed multiplex PCR- and antibody-based microbead assays using the Luminex technology. Using the Luminex assays, we were capable of serotyping 11 STEC isolates that were previously determined untypeable for the O antigen by conventional methods using antisera. Except for 14 isolates, results from the 2 Luminex assays agreed.

An energetic evaluation of dissolution corrosion capabilities of liquid metals on iron surface.

Using first principles calculations, dissolution corrosion of liquid metals on iron surfaces has been investigated by calculating adsorption energies of metal atoms in the liquid phase on the surface and escape energies of surface Fe atoms. The adsorption energies, characterizing the stability of the adsorbed atoms on the investigated surfaces, show that Bi is more stable than Pb and Au. The escape energies, representing the energy required for an Fe atom to escape from the surface, show that adsorbed Pb makes surface Fe atoms escape more easily than Bi and Au. The combination of adsorption energy and escape energy indicates that the corrosion capabilities of liquid metals decrease in the order Bi > Pb > Au. This is further proved by the investigation of surface properties, such as inter-layer distance, magnetic momentum and charge density difference. The results are consistent with experimental results that Fe can be corroded more severely in Bi than in Pb. In the case of liquid alloys, chemical proportions of compositions are incorporated to evaluate the corrosion capabilities of Pb-Bi eutectic (LBE) and Pb-Au eutectic (LGE). It is found that LBE has more severe corrosion capability than LGE. The energetic calculation is further developed in evaluating the effect of alloying elements in popular steels on the dissolution corrosion. The results indicate that Si, V, Nb and Mo may mitigate the dissolution corrosion of martensite steels in liquid Pb, Bi and Au.

Exploring the temporal-spatial characteristics and determinants of high-quality development of city clusters in China.

The complexity, severity, and uncertainty of the international situation have prompted the development of city clusters to focus more on resilience and the building of infrastructures and safeguards. Chinese-style modernization proposes a new realization path for the high-quality development (HQD) of city clusters, based on which an evaluation system for HQD indicators of city clusters is constructed. We also measured the HQD levels of 19 city clusters from 2011 to 2021 and analyzed their spatial differentiation characteristics, agglomeration evolution characteristics, and influencing factors by using kernel density, standard deviation ellipse, Moran's index, geographic detector, and geographically weighted regression. The study revealed that (1) the overall level of HQD of China's city clusters shows a trend of continuous growth, and there is obvious polarization in the high quality of city clusters in different regions. (2) The spatial distribution of HQD in city clusters decreased in the "East, Center and West" direction, but the spatial patterns of "Southeast highlighting" and "Northwest rising" became more obvious. (3) The HQD of city clusters shows obvious spatial agglomeration characteristics and overall presents a spatial pattern of "hot in the east and cold in the west", with the scope of the cold spot area gradually shrinking, and the hot spot area tends to spread outward, with mature city clusters at the core. (4) The influencing factors of HQD in Chinese city clusters are diverse, with financial levels, digital economics, human capital and green innovations having decreasing influence on HQD in city clusters but showing an obvious two-factor enhancement trend, with financial levels being able to effectively stimulate the driving potential of other factors. Financial levels can effectively stimulate the driving potential of other factors. (5) The coefficients of the driving factors affecting the HQD of city clusters vary significantly spatially, with human capital, financial levels and green innovations showing a north‒south hierarchical banded distribution of "high in the south and low in the north", and digital economy shows an east-west hierarchical belt distribution of "high west and low east". Based on the above conclusions, the realization path of accelerating the HQD of China's city clusters is proposed by optimizing the functional division of labor of the city clusters, giving full play to the comparative advantages of the hinterland city clusters, and relying on the high level of the city clusters for opening up.