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Coughing as the main symptom of a chronic cough is a persistent respiratory disease that poses a substantial challenge to Western medicine and traditional Chinese medicine (TCM). The Large Intestine and Lung are intimately related, impacting each other's pathology and physiology, according to TCM theory. This complex interaction emphasizes how crucial it is to treat both organs at the same time while treating cough to support gut health. This viewpoint is further supported by current research, which demonstrates the connections between the Large Intestine and the Lung through a variety of pathways, including neurological pathways, mucosal immune system modulation, gas exchange dynamics, and microbiota composition. To clarify the mechanisms and practical applications of treating chronic cough by concurrently targeting the Lung and Intestine in TCM, this paper merges ideas from TCM theory and empirical research in Western medicine. This provides insightful information for efficiently managing chronic cough.

Summary: Background: Hospitals in China are classified into tiers (1, 2 or 3), with the largest (tier 3) having more equipment and specialist staff. Differential health insurance cost-sharing by hospital tier (lower deductibles and higher reimbursement rates in lower tiers) was introduced to reduce overcrowding in higher tier hospitals, promote use of lower tier hospitals, and limit escalating healthcare costs. However, little is known about the effects of differential cost-sharing in health insurance schemes on choice of hospital tiers. Methods: In a 9-year follow-up of a prospective study of 0.5 M adults from 10 areas in China, we examined the associations between differential health insurance cost-sharing and choice of hospital tiers for patients with a first hospitalisation for stroke or ischaemic heart disease (IHD) in 2009–2017. Analyses were performed separately in urban areas (stroke: n = 20,302; IHD: n = 19,283) and rural areas (stroke: n = 21,130; IHD: n = 17,890), using conditional logit models and adjusting for individual socioeconomic and health characteristics. Findings: About 64–68% of stroke and IHD cases in urban areas and 27–29% in rural areas chose tier 3 hospitals. In urban areas, higher reimbursement rates in each tier and lower tier 3 deductibles were associated with a greater likelihood of choosing their respective hospital tiers. In rural areas, the effects of cost-sharing were modest, suggesting a greater contribution of other factors. Higher socioeconomic status and greater disease severity were associated with a greater likelihood of seeking care in higher tier hospitals in urban and rural areas. Interpretation: Patient choice of hospital tiers for treatment of stroke and IHD in China was influenced by differential cost-sharing in urban areas, but not in rural areas. Further strategies are required to incentivise appropriate health seeking behaviour and promote more efficient hospital use. Funding: Wellcome Trust, Medical Research Council, British Heart Foundation, Cancer Research UK, Kadoorie Charitable Foundation, China Ministry of Science and Technology, and National Natural Science Foundation of China.

Abstract Introducing superconductivity into two-dimensional (2D) films with nontrivial topology has been intensively pursued as one of the feasible scenarios to realize 1D topological superconductor. Prevailing endeavors mostly exploit the external gating or proximity effect of a traditional superconductor, by which the critical temperatures ( T c $T_{\mathrm{c}}$ ) are limited to several Kelvin range. Here, we report on the discovery of interface-enhanced superconductivity in monolayer 1T′-MoTe2 film. A thermally driven phase transition from Mo6Te6 nanowires to 1T′-MoTe2 films, grown on SrTiO3(001) surface by the molecular beam epitaxial methods, is demonstrated. A combined study of scanning tunneling microscopy/spectroscopy, electrical transport and magnetization measurements indicates the T c $T_{\mathrm{c}}$ of MoTe2 film is around 30 K, two orders of magnitude larger than its 3D counterpart crystal. This study shows that interfacial engineering is an efficient way to tune monolayer 1T′-MoTe2 film into superconducting states, and thus may pave the way toward higher- T c $T_{\mathrm{c}}$ 1D intrinsic topological superconductivity.

Background Accumulating studies demonstrated that resistance of colon cancer (CC) to 5-fluorouracil (5-FU) contributes to adverse prognosis. We investigated how Kruppel-like factor 4 (KLF4) affected 5-FU resistance and autophagy in CC cells. Methods KLF4 expression and its downstream target gene RAB26 in CC tissues was analyzed by bioinformatics analysis, and the effect of abnormal KLF4 expression on prognoses of CC patients was predicted. Luciferase reporter assay detected the targeted relationship between KLF4 and RAB26. The viability and apoptosis of CC cells were analyzed by CCK-8 and flow cytometry. The formation of intracellular autophagosomes was detected by confocal laser scanning microscopy and immunofluorescence staining. The mRNA and protein levels were assayed by qRT-PCR and western blot. A xenograft animal model was constructed to verify the function of KLF4. Rescue assay was employed to verify whether KLF4/RAB26 could affect 5-FU resistance in CC cells through autophagy. Results KLF4 and RAB26 were lowly expressed in CC. KLF4 correlated with patients’ survival. KLF4 was down-regulated in 5-FU resistant CC cells. KLF4 overexpression suppressed the proliferation and 5-FU resistance of CC cells, and inhibited LC3 II/I expression and autophagosome formation. Autophagy activator Rapamycin or sh-RAB26 treatment reversed the impact of KLF4 overexpression on 5-FU resistance. In vivo assay verified that KLF4 inhibited 5-FU resistance in CC cells. Rescue experiments revealed that KLF4 targeted RAB26 to inhibit CC cell autophagy, resulting in decreasing the resistance to 5-FU. Conclusion KLF4 strengthened the sensitivity of CC cells to 5-FU by targeting RAB26 to restrain autophagy pathway.

Background and objective SMARCA4-deficient non-small cell lung cancer (SMARCA4-dNSCLC) is a rare primary lung malignancy. These diseases are not listed separately in the 2021 World Health Organization (WHO) classification of lung neoplasms, but they have special morphological, immunophenotypic and molecular genetic characteristics. This study aims to improve understanding of SMARCA4-dNSCLC by discussing the clinicopathological features, diagonosis and differential diagnosis of the disease. Methods The clinical and imaging data of 9 cases of SMARCA4-dNSCLC diagnosed in Shanghai Changhai Hospital from January 2020 to March 2022 were collected. The clinicopathological features were analyzed by histological and immunohistochemical staining, and the literature was reviewed. Results The median age of 9 patients was 65 years old. Six men were smokers. The average maximum diameter of tumor was 3.3 cm. Six cases had been metastasized. The imaging showed that it was an infiltrating mass with unclear boundary and 3 cases invaded the pleura. Nine cases were diagnosed as SMARCA4-dNSCLC, which mainly showed three pathological forms including classic lung adenocarcinoma, mucinous adenocarcinoma and poorly differentiated carcinoma. Poorly differentiated tumor cells are epithelioid, syncytial or rhabdomyoid, the cytoplasm was rich, the cytoplasm could be completely transparent to eosinophilic, eosinophilic globules or small abscesses could be seen, showing solid flakes, with more inflammatory cells and flake necrosis in the stroma. Immunohistochemistry showed that SMARCA4 was negative in all cases and eight cases demonstrated cytokeratin 5.2 (CAM5.2) and cytokeratin 7 (CK7) was diffusely strongly positive, p40 was negative, thyroid transcription factor-1 (TTF-1) was negative in 6 cases, partially positive in 2 cases and diffusely positive in 1 case. Conclusion SMARCA4-dNSCLC is a rare subtype of lung cancer with complex and diverse pathological morphology. The characteristic of immunohistochemical phenotype can assist in the diagnosis.

Diabetic ischemic wound treatment remains a critical clinical challenge. Neovascularization plays a significant role in wound healing during all stages of the tissue repair process. Strategies that enhance angiogenesis and neovascularization and improve ischemic pathology may promote the healing of poor wounds, particularly diabetic wounds in highly ischemic conditions. We previously identified a cyclic peptide LXW7 that specifically binds to integrin αvβ3 on endothelial progenitor cells (EPCs) and endothelial cells (ECs), activates vascular endothelial growth factor (VEGF) receptors, and promotes EC growth and maturation. In this study, we designed and synthesized a multi-functional pro-angiogenic molecule by grafting LXW7 and collagen-binding peptides (SILY) to a dermatan sulfate (DS) glycosaminoglycan backbone, named LXW7-DS-SILY, and further employed this multi-functional molecule to functionalize collagen-based extracellular matrix (ECM) scaffolds. We confirmed that LXW7-DS-SILY modification significantly promoted EPC attachment and growth on the ECM scaffolds in vitro and supported EPC survival in vivo in the ischemic environment. When applied in an established Zucker Diabetic Fatty (ZDF) rat ischemic skin flap model, LXW7-DS-SILY-functionalized ECM scaffolds loaded with EPCs significantly improved wound healing, enhanced neovascularization and modulated collagen fibrillogenesis in the ischemic environment. Altogether, this study provides a promising novel treatment to accelerate diabetic ischemic wound healing, thereby reducing limb amputation and mortality of diabetic patients.

BackgroundColorectal surgery is associated with a high risk of surgical site infection (SSI). In March 2017, we developed an intervention, called “PRESS”, with the aim of reducing colorectal superficial SSI. This study assessed the effect of the new intervention in reducing the rates of superficial SSI in colorectal surgery.MethodsThis study was a retrospective review of 312 PRESS+ patients compared to 171 historical control PRESS− patients who were 18 years of age or older and underwent elective colorectal surgery with clean-contaminated wounds from January 2015 to June 2020. In the PRESS+ groups, we pressed the incision downward hard with clean gauze after the interrupted suturing of the skin. Propensity score matching with 15 variables was performed in a 1:1 ratio to reduce selection bias. Univariate analysis and multivariate analysis were performed to identify risk factors associated with SSI.ResultsThe characteristics of the PRESS+ (n = 160) and PRESS− (n = 160) groups were well balanced after propensity score matching. The PRESS+ group had a lower superficial SSI rate (1.9% vs. 6.9%, P = 0.029) and a lower overall SSI rate (2.5% vs. 10.0%, P = 0.006) than the PRESS− group. Furthermore, multivariate analysis showed that the incisional press was an effective protective factor for superficial SSI (adjusted odds ratio = 0.215, 95% confidence interval = 0.057–0.818, P = 0.024). In addition, female sex (P = 0.048) and blood transfusion (P = 0.011) were demonstrated to be independent risk factors for superficial SSI.ConclusionThe incisional press after suturing is a simple, costless, and effective intervention in reducing superficial incisional SSI.

The development and supporting technologies of high-strain line pipe steel and pipes used for oil and gas pipeline projects in special geological environments (such as earthquake zone, landslide zone, mine goaf and subsidence zone) is one of the international research hotspots, and it is also a major problem that China's major oil and gas pipeline projects have to solve. Focusing on a series of key technical difficulties in the research & development and application of high-strain line pipe steel and pipes, a number of theoretical and technological innovations have been achieved after more than ten years of joint researches. And the main achievements are as follows. First, the method of applying many different parameters (e.g. stress ratio, yield to tensile ratio and uniform elongation (UEL)) to comprehensively characterize and evaluate the deformation behavior of steel pipes is proposed, and a technical index system and standard for the new products of X70HD/X80HD high-strain line pipe steel and pipes are established. Second, a complete set of X70HD/X80HD steel plate manufacturing technology is researched and developed, and the high-strain steel plate with the properties of low yield to tensile ratio, high uniform elongation, high stress ratio and high strength and toughness is worked out. Third, the X70HD/X80HD JCOE and UOE high-strain longitudinal submerged arc welded pipe manufacturing technology is researched and developed, reasonable performance matching between a steel pipe and a welding seam is realized, and the performance deterioration in the process of forming, welding, diameter-expending and thermal coating is solved. Fourth, a full-scale test device of steel pipe internal pressure + bending large deformation is independently developed, and the physical simulation based steel pipe deformation test technology is formed. X70HD/X80HD high-strain line pipe steel and pipes have been applied in the West to East Gas Pipeline, the China–Myanmar Pipeline and other major gas pipeline projects in the large scale, and their application effects are remarkable. To satisfy the new needs of pipeline construction and long-term safe operation under complex geological conditions, some suggestions were put forwarded, such as developing new methods of strain-based pipeline design, and researching and developing or improving the supporting technologies suitable for the higher matching requirements of girth weld, such as welding methods, welding materials, welding processes, and girth weld performance quality and defect control requirements.

Considering the low indoor positioning accuracy and poor positioning stability of traditional machine-learning algorithms, an indoor-fingerprint-positioning algorithm based on weighted k-nearest neighbors (WKNN) and extreme gradient boosting (XGBoost) was proposed in this study. Firstly, the outliers in the dataset of established fingerprints were removed by Gaussian filtering to enhance the data reliability. Secondly, the sample set was divided into a training set and a test set, followed by modeling using the XGBoost algorithm with the received signal strength data at each access point (AP) in the training set as the feature, and the coordinates as the label. Meanwhile, such parameters as the learning rate in the XGBoost algorithm were dynamically adjusted via the genetic algorithm (GA), and the optimal value was searched based on a fitness function. Then, the nearest neighbor set searched by the WKNN algorithm was introduced into the XGBoost model, and the final predicted coordinates were acquired after weighted fusion. As indicated in the experimental results, the average positioning error of the proposed algorithm is 1.22 m, which is 20.26–45.58% lower than that of traditional indoor positioning algorithms. In addition, the cumulative distribution function (CDF) curve can converge faster, reflecting better positioning performance.

The authors investigate 3D-printed tips, based on controlled microstructural architectured materials, as probes for shear-mode atomic force microscopy. They demonstrate that the tailored stiffness and energy-absorbing behaviour of the material are beneficial for improving image quality.

Ideal single-cell viscometer has remained unachieved, leaving a gap in current palette of single-cell nanotools. Information of single-cell viscosity could contribute to our knowledge of fundamental biological processes, e.g., mass diffusion, biochemical interaction, and cellular responses to many diseases and pathologies. Although advances have been made to this end, existing methods generally suffer from limitations, e.g., low spatiotemporal resolution. Here, we describe a high spatiotemporal iontronic single-cell viscometer that operates upon a patch clamp integrated with double-barreled nanopores separated by a septum of ca. 32 nm. The system enables reversible electroosmotic manipulation of the adjacent small fluid bridging two nanopores, the viscous alternation of which could be sensitively monitored by the ionic responses. In practical cellular studies, significantly, our findings reveal not only the less deviated medium viscosities than those of lysosomes and mitochondria but also the highest viscosities in the near-nuclear region than those of mitochondrion-dense and lysosome-dense regions. This work has provided an accessible single-cell viscometer and enriched the armory of single-cell nanotools.

Separation of strontium isotope is an important and challenging work. Herein, a novel method was developed for preparing of acid-resistance crown ether pillared zirconium phosphonate (D-AM-ZrP) by a directly one step reacting 4-amino-benzo-18-crown-6 phosphonic acid with zirconium other than the conventional complicated intercalation methods. By suitable control of the ratio of phosphoric acid to phosphonic acid, the D-AM-ZrP sample with perfect flower-like and well-ordered interlayer structure was obtained. More importantly, D-AM-ZrP exhibited high adsorption capacity and excellent selectivity for extracting Sr2+ from complicated system including high concentrated HNO3 media and competing ions, and the maximum adsorption capacity for Sr2+ (136.42 mg·g-1) was better than that of pure α-ZrP (28.17 mg·g-1) at weak acidic media, which could mainly be attributed to the presence of the strong chemical interaction between the intercalated molecule and material, resulting in its excellent stability and abundant adsorption sites.

In this work, the Co-Ni basic carbonate nanowires were in-situ grown on carbon nanotube (CNT) network through a facile chemical bath deposition method, which could be further converted into active hydroxide via cyclic voltammetry strategy. A series of carbonate nanowire/nanotube with different Co/Ni ratio revealed the different growth status of the nanowires on CNT network. The nanostructures of the as-synthesized samples were examined via powder X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) techniques. The Co/Ni ratio of the carbonate largely affected the size of the nanowires, that the low Co/Ni ratio was beneficial for thin nanowire formation and the nanowires loading on CNT network. Subsequently, the electrochemical performance of the Co-Ni basic hydroxides was studied in a three-electrode test system. The nanowires with low Co/Ni ratio 1/2 can form nanowire array on individual CNTs, which exhibited better electrochemical capacitive performance than the composite network with high Co/Ni ratio nanowires after electrochemical activation. The addition of Co enhanced the rate performance of the hydroxide/CNT, especially improved the long cycle stability largely compared to the rate performance of pure Ni converted hydroxide/CNT composite film reported by our previous research. This result is valuable for the design of inorganic electrochemical active composites based on conductive networks for energy conversion/storage applications.

Vascular endothelial growth factor receptor 2 (VEGFR-2) and neuropilin-1 (NRP-1) are two prominent antiangiogenic targets. They are highly expressed on vascular endothelial cells and some tumor cells. Therefore, targeting VEGFR-2 and NRP-1 may be a potential antiangiogenic and antitumor strategy. A7R, a peptide with sequence of Ala-Thr-Trp-Leu-Pro-Pro-Arg that was found by phage display of peptide libraries, can preferentially target VEGFR-2 and NRP-1 and destroy the binding between vascular endothelial growth factor 165 (VEGF165) and VEGFR-2 or NRP-1. This peptide is a new potent inhibitor of tumor angiogenesis and a targeting ligand for cancer therapy. This review describes the discovery, function and mechanism of the action of A7R, and further introduces the applications of A7R in antitumor angiogenic treatments, tumor angiogenesis imaging and targeted drug delivery systems. In this review, strategies to deliver different drugs by A7R-modified liposomes and nanoparticles are highlighted. A7R, a new dual targeting ligand of VEGFR-2 and NRP-1, is expected to have efficient therapeutic or targeting roles in tumor drug delivery. Keywords: A7R, Vascular endothelial growth factor receptor 2, Neuropilin-1, Antiangiogenesis, Tumor angiogenesis imaging, Drug delivery

Abstract Background Recent studies have shown that CD103 is an important marker for tissue-resident memory T cells (TRM) which plays an important role in anti-infection. However, the role of CD103+ TRM was not elucidated in the progress of S. japonicum infection induced disease. Methods 6–8 weeks old C57BL/6 mice were infected by S. japonicum. Mice were sacrificed and the lungs were removed 5–6 weeks after infection. Immunofluorescent staining and Q-PCR were performed to identify the expression of CD103 molecule. Single cellular populations were made, percentages of CD103 on both CD4+ and CD8+ T lymphocytes were dynamical observed by flow cytometry (FCM). Moreover, the expression of memory T cells related molecules CD69 and CD62L, T cell function associated molecules CD107a, IFN-γ, IL-4, IL-9, and IL-10 were compared between CD103+ CD4+ and CD8+ T cells by FCM. Results CD103+ cells were emerged in the lung of both naive and S. japonicum infected mice. Both the percentage and the absolute numbers of pulmonary CD4+ and CD8+ cells were increased after S. japonicum infection (P

Abstract The hot deformation characteristics of Nickel‐based corrosion resistant alloy was studied in the temperature range of 1050°C to 1200°C and the strain rate range of 0.001 to 0.1 second−1 by employing hot compression tests. The results show that the peak stress increases with decreasing temperature and increasing strain rate, and the apparent activation energy is about 409 kJ/mol. Based on the Avrami equation through using the critical strain (εc) and the strain for 50% DRX (ε0.5), a kinetic model for dynamic recrystallization (DRX) was established, where the model parameters could be obtained using the modified Zener‐Hollomon parameter (Z*). Applying the model, the predicted value of the steady‐state strain (εss) and the strain for maximum softening rate (εm) agree well with the experimental results. Accordingly, the relationship between εm and ε0.5 is established, which is mainly dependent on the Avrami exponent (n). When n

Radix Pseudostellariae, the root of Pseudostellaria heterophylla (Miq.) Pax, has historically been used as medicine food homology plant for thousand years in China. Herein, PHP, a water soluble polysaccharides from Radix Pseudostellariae, was isolated and identified by HPGPC, GC–MS, FT-IR, and scanning electron microscopy. Results showed that PHP (Mw = 1.07 × 104 kDa) mainly consisted of rhamnose, galacturonic acid, glucose, galactose, arabinose, fucose in a molar ratio of 0.055:0.208:0.306:0.055:0.753:0.594:0.014. It contained nine types of linkages, including → 6)-Galp(A)-(1→, →6)-Galp-(1→, →3)-Arap-(1→, Galp-(1→, →3)-Rhap-(1→, Glup-(1→, →3)-Araf-(1→, →2,3,4)-Manp-(1 → and → 3,4)-Fucp-(1→. Morphology analysis showed that PHP took flakes with irregular size and properties. It significantly activated TLR4 in vivo. In DSS induced mouse-acute-colitis molds, PHP90-3a improved and the colon length was on ulcerative in mice. the production of IL-1β and TNF-α were decreased. The intestinal mucosa was more complete, there were much more goblet cells, and the infiltration of lymphocytes and neutrophils was significantly decreased. The content of SCFA increased, whereas acetic acid, propionic acid and butyric acid increased. 16S RNA sequencing and OTUs cluster analysis revealed that Bacteroides and Pseudomonas were the dominant intestinal flora. The analysis of community thermogram showed that the abundance ratio of Pseudomonas and Bacteroides (F/B) was significantly lower than the dominant group and significantly inhibited the levels of Verrucomicrobia and Akkermansia. Altogether, the data suggested that the anti-inflammatory activity might be implicated with the activation of TLR4 signal pathways, reducing IL-1β and TNF-α, and the regulation of mucosal immunity and the composition of intestinal flora.

Many countries, especially China, have extensively promoted liquefied natural gas (LNG) to replace diesel in heavy-duty vehicles for to achieve sustainable transport aims, including carbon peaks and neutrality. We developed a life-cycle calculation model for environmental load differences covering vehicle and fuel cycles to comprehensively compare the LNG tractor-trailer and its diesel counterpart in China on a full suite of environmental impacts. We found that the LNG tractor-trailer consumes less aluminum but more iron and energy; emits less nitrogen oxide, sulfur oxide, nonmethane volatile organic compounds, and particulate matter but more greenhouse gases (GHG) and carbon monoxide (CO); and causes less abiotic depletion potential, acidification potential, and human toxicity potential impacts but more global warming potential (GWP) and photooxidant creation potential (POCP) impacts. Poor fuel economy was found to largely drive the higher life-cycle GHG and CO emissions and GWP and POCP impacts of the LNG tractor-trailer. Switching to the LNG tractor-trailer could reduce carbon dioxide by 52.73%, GWP impact by 44.60% and POCP impact by 49.23% if it attains parity fuel economy with its diesel counterpart. Policymakers should modify the regulations on fuel tax and vehicle access, which discourage improvement in LNG engine efficiency and adopt incentive polices to develop the technologies.

Nuclear factor-kappa B (NF-κB) signaling is regulated by ubiquitin to maintain immune homeostasis. Here the authors show that a peptidylprolyl isomerase, CYPJ, blocks TAB2/3 or LUBAC ubiquitin chain sensing and suppress NF-κB activation, with CYPJ-deficiency leading to susceptibility to inflammatory stimuli.

Summary: The transcription factor IRF3 is phosphorylated in response to viral infection, and it subsequently forms a homodimer and translocates into the nucleus to induce the transcription of genes important for antiviral immunity, such as type I interferons (IFNs). This multistep process is essential for host defense against viral infection, but its regulation remains elusive. Here, we report that the EF-hand protein calmodulin-like 6 (CALML6) directly bound to the phosphorylated serine-rich (SR) region of IRF3 and impaired its dimerization and nuclear translocation. Enforced CALML6 expression suppressed viral infection-induced production of IFN-β and expression of IFN-stimulated genes (ISGs), whereas CALML6 deficiency had the opposite effect. In addition, impaired IFN-β and ISG expression in bone-marrow-derived macrophages and tissues of CALML6 transgenic mice promoted viral replication. These findings identify a phosphorylation-dependent negative feedback loop that maintains the homeostasis of antiviral innate immunity. : Wang et al. report that the EF-hand protein CALML6 binds to the phosphorylated serine-rich region of IRF3 to impair IRF3 dimerization and nuclear translocation, thereby attenuating the transcription of type I interferons. This work identifies a negative feedback regulatory loop that maintains homeostasis during antiviral innate immunity. Keywords: calmodulin-like 6, CALML6, interferon regulatory factor 3, IRF3, phosphorylation, dimerization, antiviral innate immunity

Toll-like receptors (TLRs) play an important role in regulating immune responses during pathogen infection. However, roles of TLRs on T cells reside in the mesenteric lymph node (MLN) were not be fully elucidated in the course of S. japonicum infection. In this study, T lymphocytes from the mesenteric lymph node (MLN) of S. japonicum-infected mice were isolated and the expression and roles of TLR2, TLR3, TLR4, and TLR7 on both CD4+ and CD8+ T cells were compared. We found that the expression of TLR7 was increased in the MLN cells of S. japonicum-infected mice, particularly in CD4+ and CD8+ T cells (P

CC chemokine receptor 4 (CCR4) has attracted much attention as a promising therapeutic drug target for CCR4+ tumor cells and Tregs. CCR4 is expressed on some tumor cells such as T‐cell acute lymphoblastic leukemia (ALL), adult T‐cell leukemia/lymphoma (ATLL), adult peripheral T cell lymphoma (PTCL) and cutaneous T cell lymphoma (CTCL). CCR4 is also expressed on majority of Tregs, mainly effector Tregs. In this study we have successfully developed three versions of diphtheria‐toxin based anti‐human CCR4 immunotoxins (monovalent, bivalent and single‐chain fold‐back diabody). Binding analysis by flow cytometry showed that all three versions of the anti‐human CCR4 immunotoxins bound to the human CCR4+ tumor cell line as well as CCR4+ human PBMC. The bivalent isoform bound stronger than its monovalent counterpart and the single‐chain foldback diabody isoform was the strongest among the three versions. In vitro efficacy analysis demonstrated that the bivalent isoform was 20 fold more potent in inhibiting cellular proliferation and protein synthesis in human CCR4+ tumor cells compared to the monovalent anti‐human CCR4 immunotoxin. The single‐chain fold‐back diabody isoform was 10 fold more potent than its bivalent counterpart and 200 fold more potent than its monovalent counterpart. The in vivo efficacy was assessed using a human CCR4+ tumor‐bearing mouse model. The immunotoxin significantly prolonged the survival of tumor‐bearing NOD/SCID IL‐2 receptor γ−/− (NSG) mice injected with human CCR4+ acute lymphoblastic leukemia cells compared with the control group. This novel anti‐human CCR4 immunotoxin is a promising drug candidate for targeting human CCR4+ tumor cells and Tregs in vivo.